Você está na página 1de 249

Pro-drugs as Novel Drug

Delivery Systems

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

Pro-drugs as Novel Drug


Delivery Systems
T . Higuchi and V . Stella, Editors

A symposiu
the Division of Medicinal
Chemistry at the 168th
Meeting of the American
Chemical Society,
Atlantic City, N . J . ,
September 10, 1974.

ACS S Y M P O S I U M SERIES

AMERICAN CHEMICAL SOCIETY


1975
WASHINGTON, D. C.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

14

Library of Congress

Data

Pro-drugs as novel drug delivery systems.


(ACS symposium series; 14)
Includes bibliographies and index.
1. Chemistry, Medical and pharmaceuticalCongresses.
2. Drug metabolismCongresses.
I. Higuchi, Takeru, 1918-

II. Stella, V . , 1946-

III. American Chemical Society. Division of Medicinal


Chemistry.

IV. American Chemical Society.

V . Title.

VI. Series: American Chemical Society. ACS symposium


series; 14.
RS421.S92 1974

615'.7

ISBN 0-8412-0291-5

75-11721
ACSMC8 14 1-245

Copyright 1975
American Chemical Society
All Rights Reserved
PRINTED I N T H E U N I T E D STATES O F AMERICA

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

ACS Symposium Series


Robert F. Gould, Series Editor

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

FOREWORD
The ACS SYMPOSIU

a medium for publishing symposia quickly in book form. The


format of the SERIES parallels that of its predecessor, ADVANCES
IN CHEMISTRY SERIES, except that in order to save time the
papers are not typeset but are reproduced as they are submitted by the authors in camera-ready form. As a further
means of saving time, the papers are not edited or reviewed
except by the symposium chairman, who becomes editor of
the book. Papers published in the ACS SYMPOSIUM SERIES
are original contributions not published elsewhere in whole or
major part and include reports of research as well as reviews
since symposia may embrace both types of presentation.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

PREFACE
'his volume contains the papers presented during the Symposium on
Pro-drugs held at Atlantic City, September 10,1974 under the sponsorship of the Division of Medicinal Chemistry. No serious effort was made
in organizing the program to provide a comprehensive treatment of the
subject matter. Rather it was hoped that the material presented would
stimulate greater interest in this chemical approach to the problem of
drug delivery.
A short review of the subject sets forth some of the basic underlying
concepts and approaches
array of antibiotics are then discussed. The remainder of the volume
details chemical and biological studies on pro-drug candidates developed
in our own laboratories.
We apologize for the limited coverage of the subject matter contained in this book. This was caused more by necessity than by choice.
At the time the program was formulated we were forced to depend
largely on projects completed or being carried out in our several facilities
or in those of our collaborators. This was in large part due to the sensitive
nature of pro-drug research programs in established drug houses and
their reluctance to publicize their early efforts in the field. In any case,
the examples have been selected to illustrate the real utility of this
approach.
I would like to take this opportunity to thank all those who took
part in the program and Naida Jimenez and her able secretarial assistants
for their help in preparing the manuscript.
A

University of Kansas

TAKERU HIGUCHI

Lawrence, Kans.
March 11, 1975

ix

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1
Pro-drugs: An Overview and Definition
V. STELLA
University of Kansas, Department of Pharmaceutical Chemistry, Lawrence, Kans. 66044

Historically the term pro-drug was first introduced by Albert (1,2)


"pro-agent" to describ
biotransformation prior to exhibiting their pharmacological effects. Albert suggested that this concept
could be used for many different purposes. For
example, in his book "Selective Toxicity" (2) he mentions that "as a means of introducing selectivity into
toxicity, the principle of latent activity has endless
possibilities." Albert himself points out that the
pro-drug approach is not new. Methenamine (I) and
aspirin (II), both synthesized in the late nineteenth
century, are examples of bioreversible derivatives of

(I)

(II)
formaldehyde and salicylic acid respectively. Use of
salicylic acid as an analgesic and anti-inflammatory
agent was somewhat limited by its corrosiveness which
was in part overcome with the use of II. Formaldehyde, although a useful topical antiseptic, could not
be used orally as a urinary tract antiseptic until i t
was converted to I and formulated in an enteric coated
tablet.
The chemical modification of drugs to overcome
pharmaceutical problems has also been termed "drug
1
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

PRO-DRUGS

l a t e n t i a t i o n . " The term was f i r s t used by Harper


(3.3!) f o l l o w i n g the suggestion of Dr. L. Golberg.
Harper defined drug l a t e n t i a t i o n "as the chemical
m o d i f i c a t i o n of a b i o l o g i c a l l y a c t i v e compound to form
a new compound which upon i n v i v o enzymatic attack
w i l l l i b e r a t e the parent compound. The chemical a l t e r a t i o n s of the parent compound are such that the
change i n physicochemical p r o p e r t i e s w i l l a f f e c t the
absorption, d i s t r i b u t i o n and enzymatic metabolism."
Kupchan et a l . ( 5 . ) , i n attempting to u t i l i z e the prodrug or drug l a t e n t i a t i o n approach f o r s o l v i n g v a r i o u s
problems, extended the d e f i n i t i o n of drug l a t e n t i a t i o n
to include nonenzymatic r e g e n e r a t i o n of the parent
compound. Regeneration takes place as a consequence
of h y d r o l y t i c , d i s s o c i a t i v
n e c e s s a r i l y enzyme
The te^ms pro-drugs, l a t e n t i a t e d drugs and b i o r e v e r s i b l e d e r i v a t i v e s have been used interchangeably.
S i n k u l a and Yalkowsky i n t h e i r review (6) s t a t e , "by
i n f e r e n c e , l a t e n t i a t i o n i m p l i e s a time l a g element or
time component involved i n regenerating the b i o a c t i v e
parent molecule i n v i v o . . . .
the term pro-drug i s
general i n that i t includes l a t e n t i a t e d drug d e r i v a t i v e s as w e l l as those substances which are converted
a f t e r a d m i n i s t r a t i o n to the a c t u a l substance which
combines with receptors."
The term pro-drug i s a catchy, generic term f o r
agents which undergo biotransformation p r i o r to exh i b i t i n g t h e i r pharmacological actions and w i l l be
used i n t h i s manuscript to describe both s p e c i f i c a l l y
designed b i o r e v e r s i b l e d e r i v a t i v e s of a troublesome
compound as w e l l as "accidents" or r e t r o s p e c t i v e prodrugs. As A l b e r t (2) s t a t e s , many pro-drugs are the
r e s u l t of "accidents" rather than f o r e s i g h t e d attempts
to overcome some p h y s i o l o g i c a l , p h y s i c a l or psychol o g i c a l b a r r i e r . For example, anthracene glycosides
e x h i b i t t h e i r l a x a t i v e a c t i o n through t h e i r aglycone
while codeine may exert i t s a c t i o n due to the format i o n of morphine (2).
The u t i l i z a t i o n of the pro-drug approach has been
growing since s c i e n t i s t s began to r e a l i z e that problems such as l a c k of s o l u b i l i t y , poor b i o a v a i l a b i l i t y
due to p o l a r i t y or " f i r s t pass" e f f e c t , or lack of
chemical s t a b i l i t y could be overcome by preparing
chemically a l t e r e d temporary transport forms of the
drug (see I I I ) .
Once the b a r r i e r to the use of the
parent compound has been overcome, these temporary
transport forms can be converted to the parent compound. This releases the transport moiety "C," which

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

DRUG
A

TEMPORARY
TRANSPORT
MOIETY
C

LINKAGE

CHI)

obviously has to be n o n t o x i c , so that the parent drug


i s f r e e to exert i t s pharmacological a c t i v i t y .
Is a s a l t or a complex of a drug a pro-drug?
The question of d e f i n i n g drug d e r i v a t i v e s as pro-drugs
can be quite c o n t r o v e r s i a l
Linkage B i s normally
thought of as a covalen
a phosphate ester l i n k a g e
such products as benzathine p e n i c i l l i n have been
viewed as pro-drugs or as examples of drug l a t e n t i a
t i o n ( 3 . , 4 . ) , yet the linkage "B between the parent
compound, p e n i c i l l i n , and the transport moiety, ben
z a t h i n e , i s e l e c t r o s t a t i c . The regeneration of p e n i
c i l l i n from benzathine p e n i c i l l i n i s simply d i s s o c i a
t i o n of t h i s poorly water s o l u b l e s a l t . A complex or
a s a l t i s a chemically defined new substance, i . e . , a
new thermodynamic e n t i t y , j u s t as a m o d i f i c a t i o n i n
v o l v i n g covalent bonding r e s u l t s i n a new chemical
substance. I f the p h y s i c a l and chemical p r o p e r t i e s of
t h i s new substance give i t unique q u a l i t i e s capable of
overcoming some undesirable b a r r i e r to the use of the
parent compound and t h i s new substance r e v e r t s to the
parent compound a f t e r overcoming t h i s b a r r i e r , then i t
i s the b e l i e f of t h i s author that no matter how t r i v
i a l the chemical m o d i f i c a t i o n may be, the new sub
stance i s a pro-drug of the parent compound. A num
ber of semantic arguments f o r and against t h i s d e f i n i
t i o n can be made and r e s e r v a t i o n s about c a l l i n g the
s a l t of a compound a pro-drug would be r e a d i l y ad
m i t t e d . However, procaine and benzathine p e n i c i l l i n
(7-11), and mafenide acetate ( s a l t not amide) f o r
t o p i c a l a p p l i c a t i o n (12-14) are examples of s p e c i f i c
s a l t forms of a parenF~compound which impart unique
and important q u a l i t i e s to the s t a b l e , prolonged, and
e f f i c i e n t r e l e a s e c h a r a c t e r i s t i c s of the parent com
pound. S i m i l a r l y , the d i s c u s s i o n l a t e r by Dr. Repta
of the water soluble gentisate complex of hexamethylmelamine w i l l demonstrate the uniqueness of t h i s com
b i n a t i o n over the parent compound, thus q u a l i f y i n g the
complex as a pro-drug of hexamethylmelamine.
!!

ff

!!

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

PRO-DRUGS

Reviews and overviews of the pro-drug concept are


numerous. Apart from the c l a s s i c reviews of Harper
(3,1) and A l b e r t (1,2), the reviews of Ariens ( 1 5 - 1 8 ) ,
Bundgaard ( 1 9 ) , S i n k u l a et a l . ( 6 , 2 0 ) , and S t e l l a 21)
should be mentioned. Each review o f f e r s a d i f f e r e n t
approach to the pro-drug concept, even though obvious
overlap does e x i s t . N o t a r i (22) > i n h i s recent t h e s i s
on pharmacokinetics and molecular m o d i f i c a t i o n ,
touches l i g h t l y on the pharmacokinetic i m p l i c a t i o n s
of the pro-drug approach, an area of study which de
serves much more a t t e n t i o n .
Rationale f o r the Use of Pro-Drugs
The awareness
s i r e d pharmacologica
a c t i o n has r e c e n t l y been reemphasized by the r e s u r
gence and growth of pharmacokinetics which i s the
study of the time course of a b s o r p t i o n , d i s t r i b u t i o n ,
metabolism and e x c r e t i o n of drugs. The c o n c e n t r a t i o n
versus time p r o f i l e of a drug, i t s metabolite i n var
ious t i s s u e s and organs, and the time p r o f i l e of the
corresponding pharmacological response has been a par
t i c u l a r l y e x c i t i n g and f r u i t f u l area i n current phar
macokinetic research ( 2 3 - 2 5 ) . The awareness that the
onset, i n t e n s i t y and d u r a t i o n of drug a c t i o n are
g r e a t l y a f f e c t e d by the p h y s i c a l and chemical proper
t i e s of the drug has promoted the emergence of various
t h e o r e t i c a l a'nd p r e d i c t i v e models f o r drug design and
evaluation ( 2 6 - 2 8 ) .
Ariens et a l . (29) point out that drug a c t i o n i n
volves three major phases: the pharmaceutical phase,
the pharmacokinetic phase, and the pharmacodynamic
phase. The pharmacodynamic phase represents the drugreceptor i n t e r a c t i o n or b i o l o g i c a l a v a i l a b i l i t y of the
drug. Scheme I shows a s i m p l i f i e d pharmacokinetic
model f o r a t y p i c a l drug e n t i t y and demonstrates that
before a drug-receptor i n t e r a c t i o n can occur the drug
must reach the t a r g e t organ i n which the drug receptor
i s l o c a t e d . A number of b a r r i e r s may l i m i t a drug s
a b i l i t y to reach a d e s i r e d t a r g e t organ and the sub
sequent receptor s i t e and these b a r r i e r s can be of
pharmacokinetic o r i g i n . To reach an e f f e c t i v e and de
s i r e d concentration of drug at the t a r g e t organ r e
quires not only the drug to be e f f i c i e n t l y absorbed
( f ^ 1 and.k]_ e i t h e r l a r g e r e l a t i v e to other r a t e con
stants or c o n t r o l l e d ) but i t a l s o i d e a l l y r e q u i r e s
that the amount of drug i n the r e s t of the body be
minimized to prevent t o x i c i t y . The pathway shown by
the broken l i n e and r a t e constants k i and k 2 would
1

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

DRUG

DOSAGE
FORM

41

Scheme I

TISSUE
COMPARTMENT

21
w

13

"31

FAT DEPOT
OR "DEEP"
COMPARTMENT

CENTRAL
COMPARTMENT

l4

kV

ELIMINATION
PRODUCT

2 - ^ e x c r e t i o n ^metabolism
other
f = f r a c t i o n of dose absorbed

where k. = r a t e constants d e f i n i n g the transport of a drug between


various components

TARGET
ORGAN

a.

PRO-DRUGS

be i d e a l . For a comprehensive d i s c u s s i o n of how the


p h y s i c a l chemical p r o p e r t i e s of drug molecules a f f e c t
various pharmacokinetic parameters, the paper by
N o t a r i (22) should be consulted. A p r i o r i , any change
i n p h y s i c a l chemical p r o p e r t i e s of a drug molecule due
to i t s conversion t o a pro-drug could obviously a f f e c t
the time p r o f i l e of the parent drug i n various compartments. For example, on the p o s i t i v e s i d e , a prodrug may be converted t o the parent compound by a spec i f i c enzyme found only i n the target organ. I f the
parent compound passes from the target organ i n t o the
c e n t r a l compartment and i s immediately e l i m i n a t e d ,
then the pro-drug w i l l have conferred t o the drug a
degree of s p e c i f i c i t y f o r the target organ
On the
other hand, i f i n attemptin
aqueous s o l u b i l i t y
g (poo
y
f i n e d as the primary source of the b i o a v a i l a b i l i t y
problem) a w e l l absorbed, water soluble pro-drug der i v a t i v e i s s y n t h e s i z e d , care must be taken to ensure
that the pro-drug r e v e r t s t o the parent compound
(Scheme I I ) . The r a t e of conversion must insure a
buildup i n concentration of the parent drug t o a l e v e l above i t s minimum e f f e c t i v e l e v e l at i t s s i t e of
a c t i o n , i . e . , ki| >> (k3 + k2) , the point being that
once the p a r t i c u l a r undesirable b a r r i e r has been overcome, r a p i d r e v e r s i o n t o the parent compound i s des i r a b l e t o minimize other complications such as the
pro-drug being metabolized t o an i n a c t i v e metabolite
or being excreted unchanged from the body. On t h i s

PARENT
DRUG

PRO-DRUG

PRO-DRUG METABOLITE
(Inactive)

DRUG METABOLITE
(Inactive)

Scheme I I

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

p o i n t , A l b e r t (2) commented that "although a d e t a i l e d


knowledge of p e r m e a b i l i t y and enzymes can a s s i s t a
s k i l l f u l designer i n f i n d i n g u s e f u l pro-agents, he
w i l l have i n mind an organism's normal r e a c t i o n of a
f o r e i g n substance i s to burn i t up f o r food."
B a r r i e r s of nonpharmacokinetic and nonpharmacodynamic o r i g i n may also play a major r o l e i n preventing a drug from reaching a d e s i r e d target organ.
R e f e r r i n g to Scheme I , i t i s obvious that there are
other b a r r i e r s (represented by the rate constant k)
i n h i b i t i n g the drug's a b i l i t y to reach the dosage form
stage. The r e j e c t i o n of a product can be due to pat h o l o g i c a l l i m i t a t i o n s such as t o x i c i t y and high i n cidences of side e f f e c t s t e r a t o g e n i c i t y e t c
Phar
maceutical l i m i t a t i o n
chemical i n s t a b i l i t y of the compound or formulation
difficulties.
Common p s y c h o l o g i c a l l i m i t a t i o n s can be
traced to the unpleasant t a s t e of a drug, pain at an
i n j e c t i o n s i t e , or cosmetic damage to the p a t i e n t .
Economic b a r r i e r s which are also important are o f t e n
overlooked. In the economic s t r u c t u r e of our s o c i e t y ,
a drug must have the p o t e n t i a l to make economic gains
f o r i t s promotor or i t w i l l not reach the market
place. Just as pro-drugs can be used to overcome
pharmacokinetic b a r r i e r s , pro-drugs have been used to
overcome nonpharmacokinetic b a r r i e r s . An unpatented,
pharmacologically a c t i v e drug with some physico-chemi c a l p r o p e r t i e s l i m i t i n g i t s usefulness may not be of
i n t e r e s t to a large company. I f , however, the b a r r i e r
to the drug s use i s s u c c e s s f u l l y removed by b i o r e v e r s i b l e chemical m o d i f i c a t i o n and the m o d i f i c a t i o n i s
patentable, the product may then have economic potential.
The design of e f f i c i e n t , s t a b l e , s a f e , p a t i e n t
acceptable and e s t h e t i c a l l y p l e a s i n g way to d e l i v e r
a drug to i t s s i t e of a c t i o n while overcoming v a r i o u s
p h y s i c a l , chemical and s o c i a l b a r r i e r s i s c e r t a i n l y an
area where the u t i l i z a t i o n of the pro-drug approach
holds great p o t e n t i a l . Figure 1 shows the types of
b a r r i e r s that have l i m i t e d the s u c c e s s f u l screening
and/or f u l l development of suspected pharmacologically
a c t i v e agents and f o r which the pro-drug approach has
proven to be s u c c e s s f u l i n overcoming.
1

A p p l i c a t i o n s of the Pro-drug Approach


I t i s not the o b j e c t i v e of t h i s overview to d i s cuss every p o s s i b l e example where the pro-drug concept
was used to overcome a problem. That would be an a r duous task. What w i l l be presented w i l l be a b r i e f

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

PRO-DRUGS

LACK OP SITE
SPECIFICITY

TOXICITY IF
ABSORBED AS
SUCH

UNSTABLE AS " A "

POOR DOCTOR AND


NURSE ACCEPTANCE
DUE TO PRAGMATIC
PROBLEMS
POOR PATIENT
ACCEPTANCE-TASTE
ODOR, PAIN AT
INJECTION SITE

NOT ABSORBED FROM


GI TRACT BECAUSE
OF POLARITY - OR
NOT ABSORBED
THROUGH BLOOD
BRAIN BARRIER OR
SKIN

FORMULATION
PROBLEMS, E.G.,
LIQUID-TABLET
FORMULATION
DESIRED

METABOLIZED AT
ABSORPTION SITE

WATER INSOLUBLE
NOT ABSORBED NOT CAPABLE OF
DIRECT IV
INJECTION

ABSORBED TOO
QUICKLY SUSTAINED RELEASE
DESIRED

Figure 1

l i s t i n g and d i s c u s s i o n of some of the more c l a s s i c


examples as w e l l as some recent developments. Hopef u l l y , t h i s approach w i l l encourage a p p l i c a t i o n of the
pro-drug concept t o current research and problem s o l v ing by others.
The pro-drug approach has apparently l e d t o a
great deal of success i n overcoming s p e c i f i c problems
associated with c e r t a i n drug molecules. Many of the
examples that w i l l be given i n t h i s review are what
may be c a l l e d f o r e s i g h t e d pro-drugs, i . e . , cases where
the s c i e n t i s t has, through the use of the knowledge of
f a c t o r s a f f e c t i n g drug absorption, d i s t r i b u t i o n , metabolism and e x c r e t i o n , designed and synthesized prodrugs with the s p e c i f i c view t o overcome some problem
associated w i t h the parent compound. At the same
time, there are many examples of " a c c i d e n t a l " prodrugs which, i n r e t r o s p e c t , have been found t o be very
u s e f u l and s i g n i f i c a n t l y superior t o the parent com-

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

pound. When the r a t i o n a l e f o r the design or use of a


p a r t i c u l a r pro-drug i s d i s c u s s e d , i t should be stated
that the current a p p l i c a t i o n of the pro-drug may not
have been the reason f o r s y n t h e s i z i n g the pro-drug i n
the f i r s t p l a c e .
Use of Pro-Drugs i n Overcoming Absorption Problems
To s t a t e that the pro-drug approach has been used
to overcome absorption problems i s r a t h e r meaningless
unless the p a r t i c u l a r absorption b a r r i e r i s d e f i n e d .
For example, a drug may be poorly absorbed from the
g a s t r o i n t e s t i n a l (GI) t r a c t , i n t o the c e n t r a l nervous
system (CNS), i n t o th
through th s k i n e t c .
because the drug i
compounds and other h i g h l y p o l a r chemicals are not
w e l l absorbed through these b a r r i e r s because the barr i e r s are l i p o i d a l i n nature. The q u a l i f y i n g s t a t e ment should be made that some h i g h l y p o l a r molecules
such as v i t a m i n s , amino acids and carbohydrates are
absorbed through these b a r r i e r s but are absorbed by
a c t i v e t r a n s p o r t . A drug may be poorly absorbed from
the GI t r a c t because of the very water i n s o l u b l e chara c t e r i s t i c s of the drug. The r a t e determining step to
absorption may become the d i s s o l u t i o n r a t e of the
drug. A l s o , a drug may apparently be poorly absorbed
i n t o general c i r c u l a t i o n as a r e s u l t of the s o - c a l l e d
" f i r s t pass" e f f e c t (30-31). The " f i r s t pass" e f f e c t
r e s u l t s from the l o s s of the drug due to metabolism of
the drug i n the GI mucosa or l i v e r i n i t s i n i t i a l passage through these organs.
Understanding the problem drugs would be an easy
task i f they could be p a r t i t i o n e d i n t o neat categor i e s . I n v a r i a b l y , poor absorption of a drug cannot be
a t t r i b u t e d to any s i n g l e f a c t o r .
To F a c i l i t a t e Passage Through L i p i d Membranes of
Drugs w i t h Poor L i p i d S o l u b i l i t y .
Catecholamines. Chemical m o d i f i c a t i o n to i n crease l i p i d s o l u b i l i t y and to f a c i l i t a t e the absorpt i o n of pharmacologically a c t i v e catecholamines from
the GI t r a c t and through the blood b r a i n b a r r i e r (BBB)
has l e d to a great deal of study. Moderate success
has been achieved to date.
D e f i c i e n c i e s of b r a i n dopamine (IV) r e s u l t i n g
from degeneration of the s u b s t a n t i a n i g r a seem to be
associated w i t h a number of symptoms of Parkinson's
disease (35-40). Therefore, attempts have been made
to r a i s e the b r a i n l e v e l s of dopamine i n p a t i e n t s

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

10

PRO-DRUGS

HO
(IV)
s u f f e r i n g from Parkinson's disease. I t has been
stated that dopamine i t s e l f cannot be used because i t
i s incapable of bein absorbed
th BBB
fact
primarily attribute
h i g h l y i o n i z e d s t a t e at p h y s i o l o g i c a l pH. A precursor
or a pro-drug of dopamine, L-Dopa (V) or L-3,4-dihydroxyphenylalanine, has repeatedly been found to be
e f f e c t i v e i n the treatment of Parkinson's disease
(41-50). L-Dopa i s absorbed from the GI t r a c t and

DOPA DECARBOXYLASE

DOPAMINE
i n t o the CNS through the a c t i v e transport mechanism
f o r amino acids (51-52). I n the CNS, Dopa decarboxylase can convert L-Dopa to the d e s i r e d dopamine. I t
has been assumed that the poor absorption of dopamine
i s due t o i t s p o l a r i t y and h i g h l y i o n i z e d s t a t e . However, r a p i d enzymatic metabolism of c a t e c h o l molecules
v i a conjugation mechanisms such as s u l f a t i o n , g l u c u r o n i d a t i o n and O-methylation c o n t r i b u t e h e a v i l y t o
the r a p i d l o s s of dopamine i f dopamine i s administered
o r a l l y , i . e . , dopamine given o r a l l y probably never
reaches the BBB. Even w i t h L-Dopa, approximately only
20% of an o r a l l y administered dose reaches general
c i r c u l a t i o n as L-Dopa since i t can be r a p i d l y conjugated, decarboxylated, O-methylated and o x i d i z e d i n
the GI t r a c t and mucosa (53-60). The degree of t h i s
so c a l l e d " f i r s t pass" e f f e c t i n any given i n d i v i d u a l

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

11

p a t i e n t i s a f u n c t i o n of the age, genetic s t r u c t u r e ,


d i e t , e t c , of the i n d i v i d u a l .
I t has been suggested that part of t h i s " f i r s t
pass" e f f e c t might be circumvented by the use of
L-Dopa esters (VI) which can be transformed to the

(VI):

R = a l k y l substituent

a c t i v e drug, L-Dopa, f o l l o w i n g absorptio (1) .


t i a l l y t h i s may seem incongruous because i t appears
that the e s t e r should have the same absorption prob
lems as dopamine i t s e l f , i . e . , VI i s a primary amine
and the catechol groups have not been protected from
metabolism. The i o n i z a t i o n of phenolic amines i n
c l u d i n g L-Dopa has r e c e n t l y been discussed by M a r t i n
(62) and others (63-66).
The i o n i z a t i o n c h a r a c t e r i s
t i c s of molecules s i m i l a r to dopamine can be depicted
schematically by Schemes I I I and IV. In these schemes
K]_ and K2 represent the normal macroscopic or observed
i o n i z a t i o n constants and k^, k 2 , 12>
21
~
croscopic i o n i z a t i o n constants. I f i t i s assumed that
only N~OH or (00) i s absorbed through l i p o i d a l memk

a n d

a r e

(00)
Scheme I I I

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

m i

12

PRO-DRUGS

^^^

* *******

^| *mmimmmfi\ OH

Scheme IV
branes, then the percentage of t o t a l drug present at
p h y s i o l o g i c a l pH of 7
(00) w i l l b
functio
f
the various microscopi
cussion of the i n t e r r e l a t i o n s h i p s of the microscopic
and macroscopic constants see references 6 2 - 6 6 ) . The
f o l l o w i n g drugs were subjected to a n a l y s i s of 35(00.)
present at p h y s i o l o g i c a l pH; tyramine ( V I I ) , t y r o s i n e
e t h y l e s t e r ( V I I I ) , epinephrine ( I X ) , dopamine ( I V ) ,
and morphine ( X ) . An estimate of , pK2, pk]_, and
pk2 f o r a t y p i c a l L-Dopa e s t e r (VI) was made and ap
proximate $(00) c a l c u l a t e d at p h y s i o l o g i c a l pH. Table
I gives the pK]_, pK2, pk]_, pk2, R(where R i s the r a
t i o (+-)/(00) and equals k]_/k2) and $(00) at physio
l o g i c a l pH f o r these compounds. As i s r e a d i l y appar
ent from t h i s t a b l e l i t t l e passive absorption from the

(VII)

(VIII)

(X)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

13

Table I
Compound

pK

PK

8.87

IV

10.63

pk

8.90

pk

10.06

7.21

9.44

VII

9.61

10.65

VIII

7.33

9.80

9.42

7.33

IX

8.66

8.31

9.51

8.87

8.45

8.76*
9.70

7.22
10.32

0.21

15

VI

$(00)

0.03

*
60.2

4.2
0.008

0.12
53.1

0.38

7.98

Roughly estimated from the data of M a r t i n ( 6 2 )


based on the e f f e c t on p K i , pK2, p k i , pk2 of est e r i f i c a t i o n of the carboxyl group of t y r o s i n e
to give t y r o s i n e e t h y l e s t e r .
GI t r a c t or through the BBB would be expected f o r IV,
V I I or IX unless some compensation f o r the small f r a c t i o n of ( 0 0 ) present i s made i n terms of increased l i p o p h i l i c i t y and/or reduced " f i r s t pass" metabolism.
The r a t h e r high f r a c t i o n of ( 0 0 ) present at physiol o g i c a l pH f o r V I , V I I I and X suggests that these compounds should have l i t t l e problem penetrating l i p o i d a l
membranes, assuming they possess s u f f i c i e n t l i p o p h i l i c i t y . Any reduced b i o a v a i l a b i l i t y of these compounds
can probably be a t t r i b u t e d to a " f i r s t pass" e f f e c t .
L a i et a l . (6l)
have synthesized esters of L-Dopa
i n an attempt to overcome the " f i r s t pass" metabolism
of L-Dopa while Anden et^ a l . (67.) have prepared the
methyl e s t e r of t y r o s i n e to help absorption and prevent decarboxylation.
Pinder suggested 0 , 0 - d i a c e t y l - ( X I ) , 0 , 0 - d i ( t r i methylsilyl)dopamine (XII) as u s e f u l pro-drugs of
dopamine capable of penetrating the BBB (68).
If i t
i s assumed that pk2> the microscopic i o n i z a t i o n constant f o r the amino group of dopamine, i s unaffected
by a c y l a t i o n or s i l y a t i o n of the hydroxy groups, $ ( 0 0 )
i s c a l c u l a t e d to be 0 . 2 2 , i . e . , a c y l a t i o n of the hydroxy groups does l i t t l e to improve the f r a c t i o n of
n e u t r a l species present at p h y s i o l o g i c a l pH. What
a c y l a t i o n may do i s protect the hydroxy groups from
being conjugated and increase the l i p o p h i l i c i t y of the
molecule.
Pinder based h i s suggestions on the r e s u l t s

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

14

PRO-DRUGS

CH C0
o

(XIII)
of C r e v e l i n g et a l . ( 9 , 7 0 ) who had shown that 3 , 4 ,
-triacetyl (XIIT, and 3 , 4 , 3 - t r i ( t r i m e t h y l s i l y l )
(XIV) d e r i v a t i v e s of norepinephrine caused prolonged
release of the parent catecholamine i n mice b r a i n s .
However, as pointed out by C r e v e l i n g et_ a l . ( 6 9 , 7 0 )
even though both X I I I and XIV r e a d i l y entered the CNS,
the d e r i v a t i v e s survived as noncatechol e n t i t i e s f o r
long periods i n the b r a i n . For example, when H 3
tagged X I I I was given I.V. to mice approximately 20$
of t o t a l b r a i n r a d i o a c t i v i t y could be a t t r i b u t e d to
catechol species while the remaining 80$ was noncatechol s p e c i e s . In the hearts of the same animals the
reverse was the case, i . e . , the d e r i v a t i v e appeared to
be q u i c k l y converted to c a t e c h o l species. This tends
to suggest that enzymatic regeneration of the n o r e p i nephrine from X I I I may not be f a c i l e i n the CNS.
Borgman et a l . (Jl)
have r e c e n t l y synthesized a s e r i e s
of 0 , 0 - d i a c e t y l d e r i v a t i v e s of v a r i o u s dopaminergic
catecholamines i n c l u d i n g dopamine. The i n a b i l i t y of
XI to antagonize oxotremorine-induced tremor i n mice
( 7 1 ) , reserpine-induced depression ( 7 1 ) or to cause
hypothermia ( 7 1 ) i n mice suggests that P i n d e r s proposal that XI should penetrate the CNS may be e r r o neous.
The use of I.V. dopamine i n the treatment of
shock ( 7 2 , 7 3 ) suggests that dopamine pro-drugs such as
XI and other 0 , 0 - d i a c y l ( 1 1 , 7 4 ) and amino a c i d amides
d e r i v a t i v e s of dopamine (75.) might provide u s e f u l ,
f

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

15

o r a l l y b l o a v a i l a b l e forms of dopamine f o r the t r e a t


ment of shock. In the treatment of shock, p e r i p h e r a l
and not CNS l e v e l s of dopamine are d e s i r e d .
Other examples of attempted chemical m o d i f i c a t i o n
intended to promote CNS absorption of amines include
the studies of V e r b i s c a r et a l . (76.) w i t h amphetamine,
B j u r u l f et a l . (77.) w i t h chlorphentermine and Kupchan
et a l . (75") w i t h normeperidine. Each study attempted
carbamoylation of the amine i n an e f f o r t to (a) i n
crease the l i p o p h i l i c i t y of the amine by preventing
the i o n i z a t i o n r e a c t i o n . P e n e t r a t i o n i n t o the CNS
from blood has been c o r r e l a t e d to l i p o p h i l i c i t y and
the c o n c e n t r a t i o n of u n d i s s o c i a t e d molecules i n the
blood ( 7 9 - 8 1 ) : and/o (b)
t th metaboli a c t i o
of monoamine oxidas
r e s u l t s w i t h amphetamine and normeperidine were mar
g i n a l . The carbamates of amphetamine d i d appear to
provide a prolonged r e l e a s e e f f e c t . The r e s u l t s of
B j u r u l f ert a l . ( 7 7 ) w i t h N-carbethoxychlorphentermine
(XV) or Oberex TSraco), a pro-drug of the anorectic
agent chlorphentermine, showed that the pro-drug had a
" r e l a t i v e l y prolonged e f f e c t which makes one dose i n
the morning apparently s u f f i c i e n t . "

CH 3
(XV)
Water soluble v i t a m i n s . Many of the water s o l u
ble vitamins such as thiamine (vitamin , XVI),
r i b o f l a v i n (vitamin B 2 , XVII) and p y r i d o x i n e (vitamin
B6, XVIII) are h i g h l y polar and a c t i v e l y absorbed
agents.
Thiamine, being a water soluble compound w i t h a
quaternary n i t r o g e n , i s poorly absorbed i n t o the CNS
(83) and poorly absorbed from the GI t r a c t ( 8 4 - 8 7 ) .
Thiamine passes through these b a r r i e r s because both
i n CNS and o r a l a b s o r p t i o n , i t i s a c t i v e l y absorbed.
However, a c t i v e absorption processes are saturable
and/or e a s i l y i n h i b i t e d . I n h i b i t i o n of the o r a l ab
s o r p t i o n of thiamine by chronic a l c o h o l consumption
has been i m p l i c a t e d i n Wernicke's encephalopathy ( 8 8 )
and i n h i b i t e d CNS absorption of thiamine has been im
p l i c a t e d i n the Leigh's disease (89.,0) Thomson

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

16

PRO-DRUGS

0
(XVII)
et a l . (88) have shown that chronic a l c o h o l consumpt i o n and long d i e t a r y d e f i c i e n c y may reduce the i n t e s t i n a l absorption of thiamine. Thiamine undergoes
a r a t h e r unusual second i o n i z a t i o n (Scheme V) t o a
t h i o l a t e i o n (XIX), i n v o l v i n g the consumption of two
moles of hydroxide i o n f o r each mole of thiamine.
D e r i v a t i z a t i o n of XIX leads t o many l i p i d s o l u b l e prodrugs o f (a) the d i s u l f i d e type, such as thiamine prop y l d i s u l f i d e (TPD, XX), thiamine t e t r a f u r f u r y l d i s u l f i d e (TTPD, XXI) and 0,0 -dibenzoylthiamine d i s u l f i d e
( X X I I ) ; (b) d i a c y l type, such as 0,S-diacetylthiamine
(DAT, X X I I I ) ; and (c) 0,S- and S-carbonate e s t e r s of
thiamine, such as 0,S-diethoxycarbonylthiamine (DECT,
XXIV). These and many other d e r i v a t i v e s have been
synthesized i n Japan since the e a r l y 1950 s (91) The
synthesis of these d e r i v a t i v e s was not n e c e s s a r i l y
aimed at p r e f e r e n t i a l GI or CNS absorption of thiamine
but was geared mainly t o the p o s s i b l e use of these
l i p i d s o l u b l e thiamine d e r i v a t i v e s as s t a b l e food add i t i v e s . The p o l i s h i n g of r i c e had l e d t o some t h i a mine d e f i c i e n c y i n Japan. Thiamine i t s e l f could not
be used as a food a d d i t i v e f o r r i c e because i t i s too
water s o l u b l e , thus e a s i l y washed from r i c e . I t i s
a l s o chemically unstable (92) and very poorly absorbed.
f

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

17

(XIX)
Scheme V
Compounds l i k e (XX-XXIV) and t h e i r homologues do
not possess a quaternized n i t r o g e n so a l l o w i n g them to
be p a s s i v e l y absorbed from the GI t r a c t . Each are
q u a n t i t a t i v e l y converted t o thiamine once i n the body
(93).
XXIII and XXIV and t h e i r homologues are converted t o thiamine by t h i o e s t e r a s e s and esterases
(93).
The d i s u l f i d e compounds (XX-XXII) and t h e i r
homologues are converted to thiamine by a d i s u l f i d e
exchange mechanism i m p l i c a t i n g g l u t a t h i o n e and g l u t a thione reductase ( 9 4 - 9 7 ) . Grode et a l . (9.8) r e c e n t l y
speculated that d i s u l f i d e thiamine pro-drugs might be
s u s c e p t i b l e to i n t e r a c t i o n with serum p r o t e i n s v i a a
d i s u l f i d e exchange r e a c t i o n and p r e c i p i t a t e antibody
formation. Their r e s u l t s show that long term dosing
of XXI i n r a b b i t s d i d not e l i c i t antibody formation.
Thomson et a l . (8J3) have presented some e x c e l l e n t
data on thiamine blood and CNS l e v e l s i n Wernicke's
encephalopathy as w e l l as lowered blood and CNS l e v e l s
of thiamine i n a l c o h o l i c s having symptoms s i m i l a r to
but not n e c e s s a r i l y s u f f e r i n g from Wernicke's disease.
Their r e s u l t s show that XX on s i n g l e o r a l dosing r e s u l t e d i n increased, and i n some cases normal, red
blood c e l l (RBC) t r a n s k e t o l a s e a c t i v i t y i n a l c o h o l i c ,
thiamine d e f i c i e n t p a t i e n t s while thiamine i t s e l f had

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

CH
^NH

C-CH CH OH
S-SR
2

(XX) : R

C H
3

(XXI) : R = -CH 2

J3H

CH-

I
CH
NH $

C-CH CH 0CR
o

0
(XXIII) : R = -CH.
(XXIV) : R = -OC H
2

C H

^ ^

^CH.

CH ^N3

CH

CH
I

I
N,

0
II
C-CH CH 0C
I
S
0
i
C-CH CH oB
2~ 2
:

2 o

CH.

CK
(XXII)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

19

only a small e f f e c t on RBC transketolase a c t i v i t y .


Six hours a f t e r o r a l a d m i n i s t r a t i o n of XX, 5 of 6 a l
c o h o l i c p a t i e n t s with Wernicke's disease and the ac
companying b i l a t e r a l rectus palsy showed complete r e -

* 24

TIME (hours)
Annals of Internal Medicine

Figure 2. Thiamine blood levels in malnourished alco


holic patients with fatty livers () and in normal subjects
(0)> given 50 mg of XX (open symbols) or 50 mg of thia
mine hydrochloride (closed symbols) (88)

mission
showing
thiamine
dose of

of the occular p a l s y , with the s i x t h p a t i e n t


an improved c o n d i t i o n . Figure 2 gives the
blood l e v e l versus time p r o f i l e f o r a 50 mg
XX compared t o an i d e n t i c a l dose of thiamine

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

20

PRO-DRUGS

given to malnourished a l c o h o l i c p a t i e n t s with f a t t y


l i v e r and normal s u b j e c t s . Figure 3 i s a comparison
of blood l e v e l and c e r e b r o s p i n a l f l u i d concentration
versus time p r o f i l e and the accompanying c l i n i c a l r e sponse f o r a group of thiamine d e f i c i e n t a l c o h o l i c s
t r e a t e d w i t h thiamine f o r 25 hours and then t r e a t e d
with an equivalent dose of XX.

46 2 4
TIME (HOURS)
9

Annals of Internal Medicine

Figure 3. Comparison of clinical and laboratory abnormalities response to 50 mg of thiamine hydrochloride followed
by XX in thiamine deficient ahoholics (88)
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

21

Overview and Definition

Subacute n e c r o t i s i n g encephalomyleopathy (SNE) o r


Leigh's d i s e a s e , a t e r m i n a l disease a f f l i c t i n g c h i l dren, has been suspected to be due to thiamine CNS
d e f i c i e n c y p o s s i b l y caused by malabsorption of t h i a mine i n t o the CNS.
I f thiamine CNS absorption i s i n h i b i t e d , the use of a p a s s i v e l y absorbed l i p i d s o l uble thiamine pro-drug may prove u s e f u l . Pincus ( 8 9 )
attempted to use XX i n a number of Leigh's disease
cases with some degree of success. Temporary remissions have been noted ( 8 9 ) . Iwasaki ( 9 9 ) has s t u d i e d

Blood

13

0 13

0 13

TIME (hours)
Vitamins (Kyoto)

Figure 4. Thiamine levels in milk and blood after


parenteral administration of 200 mg of modified thiamine compounds given S.C. to goats (99)

the absorption of XXIII and XX i n t o a l i p i d depot


(goat milk) a f t e r subcutaneous (S.C.) i n j e c t i o n of
these d e r i v a t i v e s to goats. Iwasaki's r e s u l t s are
shown i n Figure 4. On the b a s i s of t h i s experiment,

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

22

PRO-DRUGS

XXIII would be expected to produce higher CNS l e v e l s


of thiamine than XX or thiamine i t s e l f , e s p e c i a l l y i n
the presence of an i n h i b i t e d thiamine CNS uptake mech
anism. A p r e l i m i n a r y c l i n i c a l i n v e s t i g a t i o n using
X X I I I , at Loyola Medical Center, Maywood, I l l i n o i s , on
a p o s s i b l e Leigh's disease case produced encouraging
r e s u l t s (100).
For a complete review of thiamine pro-drugs, the
paper of Kawasaki (93.) should be consulted. For a
summary of various s y n t h e t i c procedures f o r preparing
various thiamine pro-drugs, the reader i s d i r e c t e d to
the paper by Matsukawa et a l . (1).
The improved o r a l
b i o a v a i l a b i l i t y of thiamine through dosing with v a r i
ous thiamine pro-drug i w e l l documented i th Jap
anese l i t e r a t u r e (91*93,99,102-113)
Chronic a l c o h o l i n g e s t i o n has a l s o been shown to
i n h i b i t the a c t i v e absorption of r i b o f l a v i n , other
a c t i v e l y absorbed water soluble vitamins (114,115)>
amino acids (116-118) and carbohydrates (116).
F a t t y a c i d esters of r i b o f l a v i n have been synthe
s i z e d by Yagi et a l . ( l u , 1 2 0 ) "to widen the a p p l i
c a t i o n of r i b o f l a v i n to pharmaceutical and n u t r i t i o n a l
f i e l d s . " Their r e s u l t s show that
2 ,3 ,4 ,5 -tetrap a l m i t a t e , - t e t r a c a p r a t e , - t e t r a b u t y r a t e and - t e t r a propionate esters of r i b o f l a v i n could be hydrolyzed
to r i b o f l a v i n and the corresponding f a t t y a c i d by
pancreatic l i p a s e .
S h i n t a n i et_ a l . (121) have shown that the d i - and
t r i p a l m i t a t e e s t e r s of pyridoxine given o r a l l y to mice
had vitamin B5 a c t i v i t y . However, i f the esters were
given i n t r a p e r i t o n e a l l y ( I . P . ) , the v i t a m i n Bg a c t i v
i t y was diminished.
Results i n r a t s confirmed the
e a r l i e r f i n d i n g s i n mice (122). I t seemed that the
palmitate esters required cleavage to pyridoxine be
fore absorption and that i n j e c t i o n of the esters r e
s u l t e d i n t h e i r incomplete conversion to the pyridox
ine. The triaminobenzoate e s t e r of pyridoxine has
a l s o been prepared (123) as a p o s s i b l e pro-drug form
of pyridoxine.
L i p o p h i l i c d e r i v a t i v e s of ascorbic a c i d , such as
-palmitoylascorbic a c i d (124,125) and 6-stearoylasc o r b i c a c i d (126,127), have been synthesized as l i p o
p h i l i c a n t i o x i d a n t s f o r nonaqueous formulations.
Various mono- and p o l y a c y l d e r i v a t i v e s of ascorbic
f

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

23

Overview and Definition

a c i d have been synthesized with the view to increase


the aqueous s t a b i l i t y of ascorbic a c i d . The weak
v i t a m i n C a c t i v i t y of the 2 , 3 5 6 - t e t r a c e t y l d e r i v a
t i v e administered o r a l l y has been noted (128) while
6-benzoyl (129-132), -stearoyl ( 1 2 6 1 2 7 ) 7 ^ - l a u r y l
(126,127) and some d i a c e t y l d e r i v a t i v e s have v i t a m i n
C a c t i v i t y e q u i v a l e n t , but not s u p e r i o r , to ascorbic
a c i d . However, as w i l l be discussed l a t e r , the 2and/or 3-acyl d e r i v a t i v e s are more chemically s t a b l e
than ascorbic a c i d i t s e l f . The l i p o p h i l i c 6-palmit o y l d e r i v a t i v e i s used as a l i p o p h i l i c a n t i o x i d a n t .
5

Nucleosides and n u c l e o t i d e s . Another group of


h i g h l y p o l a r , poorly l i p o p h i l i c molecules with r e s u l
t i n g poor p e r m e a b i l i t
analogs of the n a t u r a purin
pyrimidin
sides (133). These compounds can i n t e r f e r e with nu
c l e i c a c i d synthesis and the synthesis of p r o t e i n s
and carbohydrates.
The r o u t i n e use of the nucleoside
analog, 6-azauridine (XXV), i n the treatment of neo
p l a s t i c diseases and p s o r i a s i s was i m p r a c t i c a l be
cause of i t s poor o r a l b i o a v a i l a b i l i t y . The poor
b i o a v a i l a b i l i t y can be a t t r i b u t e d to the poor perme
a b i l i t y c h a r a c t e r i s t i c s of XXV and/or metabolism of
0

(XXV): R = -H
(XXVI): R = - C O C H 3
(XXVII): R = -COC^H

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

24

PRO-DRUGS

XXV during the absorption process. The synthesis of


various e s t e r s of XXV such as 2 , 3 , 5 ' - t r i a c e t y l (XXVI), and 2 , 3 , 5 - t r i b e n z o y l - 6 - a z a u r i d i n e (XXVII) as
w e l l as other mono- and p o l y a c y l d e r i v a t i v e s was carr i e d out i n an e f f o r t to obtain an o r a l l y b i o a v a i l a b l e
form of XXV (134-139). XXVI on i n j e c t i o n i n p a t i e n t s
s u f f e r i n g from various n e o p l a s t i c diseases (140) was
found to be excreted i n the u r i n e as 6-azauridine (2911%) and monoacetyl-6-azauridine
(4-19?). The t r e a t ment of p s o r i a s i s with o r a l doses of XXVI of 250
mg/Kg/day proved s u c c e s s f u l ( l 4 l ) . XXVI given o r a l l y
to r a t s showed a n t i f e r t i l i t y p r o p e r t i e s s i m i l a r to XXV
w i t h the added advantage that XXVI was o r a l l y absorbed
!

(142).

Welch (134) ha
o r a l l y every 8 hours and
s completely absorbed. O
o r a l dosing, XXVI i s excreted 80$ as XXV and 11% as i t s
5 - a c e t y l d e r i v a t i v e w i t h only traces of XXVI excreted.
O r a l l y administered XXVI caused the same c l i n i c a l e f f e c t s as a molar equivalent dose of XXV given I.V.
The poor o r a l b i o a v a i l a b i l i t y of the n u c l e o s i d e ,
p s i c o f u r a n i n e , (XXVIII), has been a t t r i b u t e d to the
b a s i c i t y of i t s 6-amino group, and i t s n o n l i p o p h i l i c
character (143). Various acetate esters of XXVIII were
prepared i n c l u d i n g t h ^ t e t r a a c e t a t e e s t e r (XXIX).
Oral
CD50 studies with S. hemolyticus i n f e c t e d mice showed
XXIX to be twice as e f f e c t i v e as the parent compound
XXVIII. Figure 5 gives human serum l e v e l s of XXVIII as
f

OR

OR

(XXVIII) :
(XXIX) :

R = -H
R = -COCH

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

25

a f u n c t i o n of time a f t e r o r a l dosing w i t h XXVIII and


XXIX. The poor b i o a v a i l a b i l i t y of XXVIII from an o r a l
dose of XXVIII was confirmed and the s u p e r i o r i t y of
XXIX as an o r a l l y a v a i l a b l e form of XXVIII demonstra
t e d . Hoeksema et a l . (143.) s t a t e that the higher s o l
u b i l i t y of XXIX r e l a t i v e to XXVIII i n chloroform (>150

>

J. -L

12
3 4 5
TIME (hours)

Biochemical and Biophysical Research Communications

Figure 5. Serum levels in humans of XXVIII as a


function of time after 1.5 g oral dosing of XXVIII
() or XXIX (O) (143)

mg/ml compared to 0.007 mg/ml), while m a i n t a i n i n g a


reasonable aqueous s o l u b i l i t y (3 mg/ml compared t o
13 mg/ml), s t r o n g l y suggested that the s u p e r i o r o r a l
a v a i l a b i l i t y of XXVIII from XXIX can be a t t r i b u t e d to
the increased l i p o p h i l i c character of XXIX.
C y c l i c 3 ,5'-adenosine monophosphate (XXX), a po
l a r n u c l e o t i d e r e g u l a t o r of g l y c o g e n o l y s i s , has been
a c y l a t e d by Pasternak et a l . (144). The d i b u t a n o y l
d e r i v a t i v e , N-2
-0-dibutanoyladenosine-3 ,5 -mono
phosphate (XXXI), given I.V. to dogs showed a greater
hyperglycemic a c t i v i t y than XXX i t s e l f . Two^other de
r i v a t i v e s , the N - b u t a n o y l (XXXII) and the N - o c t a n o y l
(XXXIII) d e r i v a t i v e s , a l s o showed s u p e r i o r and pro1

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

26

PRO-DRUGS

NHR-

?
(XXX):
(XXXI):
(XXXII):
(XXXIII):

R
R
R
R

OR
1

= R = -COC H
=~COC H , R2 = -H
= -COC^H^, R = -H
3

longed hyperglycemic a c t i v i t y compared t o XXX. Posternak et a l . (144) a t t r i b u t e d t h i s g r e a t e r a c t i v i t y


to increased entrance i n t o c e l l s and/or the r e s i s t a n c e
of the d e r i v a t i v e s t o i n a c t i v a t i o n by phosphodiester
ases .
The a c e t a t e , formate and propionate e s t e r s o f 9(-D-arabinofuranosyl)adenine have been synthesized as
o r a l l y a v a i l a b l e forms of the parent drug (145) . Adamantoyl e s t e r s of v a r i o u s deoxyribonucleosides ( s p e c i
f i c a l l y the 5 e s t e r s ) have been prepared by Gerzon et
a l . (146). Although the authors a t t r i b u t e the a c t i v
i t y of the 5 -adamantoyl e s t e r s t o the i n t a c t e s t e r ,
the p o s s i b i l i t y that the esters were a c t i n g as pro
drugs of the parent nucleoside was not excluded.
f

Other p o l a r compounds. The large d i f f e r e n c e be


tween e f f e c t i v e o r a l and I.V. doses of many quaternary
ammonium drugs has been a t t r i b u t e d to the incomplete
o r a l absorption of quaternary compounds (147>148).
The o r a l absorption of quaternary ammonium compounds
from the GI t r a c t has always presented a problem. At
l e a s t one quaternary compound, thiamine, has been
shown t o be a c t i v e l y absorbed (84).

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

27

Levine et_ a l . (149) were able to show that i n t r a


molecular c y c l i z a t i o n s could be used to overcome t h i s
problem. Table I I shows four compounds. Compound
XXXV under p h y s i o l o g i c a l conditions found i n the p l a s
ma and i n t e s t i n a l t r a c t i s converted to XXXIV, the
quaternary compound, v i a an i n t r a m o l e c u l a r nucleop h i l i c r e a c t i o n . S i m i l a r l y , XXXVII i s converted to
XXXVI. The absorption f i g u r e s quoted i n Table I I were
from i n s i t u i n t e s t i n a l loop experiments and do not
r e f l e c t the concentration of quaternary compounds ac
t u a l l y appearing i n the blood stream. As can be seen
TABLE I I
Compoun
Hours

CH.

CH

"(CH )

Ik

(XXXIV)
CH

CH_
I
( C H2'5) - N

Br ( C H ) - N
0

( C H ^ Br

56

(XXXV)
CH,

CH,

(CH )
2

16.8

(XXVI)
CH
CH.
I 3,
I 3
C I ( CH ) ^ - N ( CH ) N ( CH ) - C l
0

21

(XXXVII)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

28

PRO-DRUGS

seen from the Table I I , the quaternary compounds were


poorly absorbed because of t h e i r low l i p i d s o l u b i l i t y
while the t e r t i a r y amine d e r i v a t i v e s were b e t t e r ab
sorbed. Studies (149) of the u r i n a r y e l i m i n a t i o n pro
ducts a f t e r dosing with the t e r t i a r y amine precursors
showed that metabolic pathways other than conversion
to the quaternary compounds were o c c u r r i n g . As a r e
s u l t , the superior absorption c h a r a c t e r i s t i c s of the
t e r t i a r y amine precursors d i d not n e c e s s a r i l y r e f l e c t
increased blood l e v e l s of quaternary compounds.
The mechanisms d e s c r i b i n g the i n t r a m o l e c u l a r cyc l i z a t i o n s of -haloalkylamines to t h e i r quaternary
analogues have been discussed by S t r e i t w i e s e r (150)
and Kusnetsov et a l (151)
Whil Levin
t a l pio
neered the p o s s i b l
of the -haloalkylamines to t h e i r quaternary analogues,
Ross and coworkers (152-157) i n a s e r i e s of s t u d i e s ,
have attempted to u t i l i z e t h i s concept more f u l l y .
Ross and Proden (152) studied the absorption and f o r
mation of XXXVIII~Trom XXXIX i n mouse b r a i n a f t e r I.P.
a d m i n i s t r a t i o n of XXXIX. The I.P. i n j e c t i o n of XXXVIII
i t s e l f d i d not give any detectable amounts of XXXVIII
i n the b r a i n , whereas I.P. a d m i n i s t r a t i o n of XXXIX r e
s u l t e d i n s u b s t a n t i a l b r a i n l e v e l s of XXXVIII. The
q u a n t i t a t i v e conversion of XXXIX to XXXVIII i n mouse
b r a i n homogenates was a l s o observed. The purpose of
the study was t o e f f e c t CNS absorption o f a quaternary
CH

CH

(XXXIX)

CH

(XXXVIII)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

29

compound by the a d m i n i s t r a t i o n of i t s t e r t i a r y -haloalkylamine precursor. The study of the e l i m i n a t i o n of


quaternary compounds from the CNS has been l i m i t e d by
absorption, i . e . , the study of e l i m i n a t i o n i s d i f f i
c u l t i f i t has never been e s t a b l i s h e d that the quater
nary compound was e f f e c t i v e l y absorbed i n the f i r s t
place. The very long e l i m i n a t i o n h a l f - l i f e o f XXXVIII
from the brains of mice, approximately 30 hours, dem
onstrates the poor e l i m i n a t i o n of i n s i t u formed p o l a r
m a t e r i a l s from the CNS. These r e s u l t s are consistent
with the f i n d i n g s of a long e l i m i n a t i o n h a l f l i f e f o r
the polar and charged i n s i t u formed acetate anion
e l i m i n a t i o n from the CNS (545).
Ross and coworker (155) hav subsequentl stud
ied the various parameter
of -haloalkylamines t o t h e i r quaternary d e r i v a t i v e s .

(XL)

(XLI)
Scheme VI
Scheme VI was the general r e a c t i o n studied. The dura
t i o n and i n t e n s i t y of l o c a l anesthesia using i n vivo
and i n v i t r o t e s t s , f o r both XL and XLI, were studied.
Chemical studies of the e f f e c t of and -x on the con
v e r s i o n of XL t o XLI and the l o c a l anesthetic a c t i v i t y
of both the t e r t i a r y analog i t s e l f and the formed qua
ternary compound suggested that the formed quaternary
compounds c o n t r i b u t e t o the d u r a t i o n of the anesthesia
(155*157). The l o c a l anesthetic e f f e c t s of XL and XLI
on the s c i a t i c nerve of guinea p i g s , i n v i v o , and
f r o g , i n v i t r o , showed that sustained l o c a l anesthetic
a c t i v i t y occurred f o r compounds where = 5 and =

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

30

PRO-DRUGS

- C l or -Br. Apparently the prolonged a c t i v i t y was


w e l l c o r r e l a t e d with the extremely slow e l i m i n a t i o n of
i H s i t u formed quaternary compound from the s c i a t i c nerve ( 1 5 7 ) and the concentration of the quaternary
compound i n the nerve. S i m i l a r studies with b r e t y l i u m
r e l a t e d d e r i v a t i v e s (153)> x y l o c h o l i n e r e l a t e d d e r i v a t i v e s ( 1 5 4 ) and troxonium r e l a t e d d e r i v a t i v e s ( 1 5 6 )
have a l s o r e c e n t l y been published.
The poor o r a l b i o a v a i l a b i l i t y of many a n t i b i o t i c s ,
such as a m p i c i l l i n ( 1 5 8 - 1 6 4 ) , erythromycin ( 1 6 5 - 1 6 7 ) *

oleandomycin (-168) and lincomycin (169*170) has been


a t t r i b u t e d t o both t h e i r p o l a r character as w e l l as
metabolism i n the GI t r a c t , GI mucosa or l i v e r during
absorption. The poo b i o a v a i l a b i l i t
f ampicilli
compared to a numbe
w i l l be discussed by Dr. S i n k u l a .
The a n t i b i o t i c , oleandomycin ( X L I I ) was found t o
have a f a i r l y broad a n i t b a c t e r i a l spectrum and t o be
OR 0 C H 0
I
II
CH CHCHCHCHC
0

3
0

CH^ ^^"2
(CH ) N
3

(XLII):
(XLIII):

R = -H
R = -COCH,

e f f e c t i v e both o r a l l y and p a r e n t e r a l l y ( 1 7 1 ) . I t was


subsequently shown that i t s t r i a c e t y l d e r i v a t i v e , t r i acetyloleandomycin ( X L I I I ) was more e f f e c t i v e o r a l l y
than the parent compound ( 1 6 8 , 1 7 2 - 1 7 5 ) This i n creased e f f e c t i v e n e s s was a t t r i b u t e d t o the greater
b i o a v a i l a b i l i t y of XLII from X L I I I than from XLII i t s e l f (See Figure 6 ) . Clemer et a l . have shown that
X L I I I has some a n t i b a c t e r i a l a c t i v i t y of i t s own
against S. aureus and S. l u t e a but that the a c t i v i t y
was only 2 5 $ that of oleandomycin free base ( 1 7 2 ) .
A f t e r i n g e s t i o n of X L I I I , XLII i s detected i n urine

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

31

along with 3-acetyloleandomycin, (major m e t a b o l i t e ) ,


1-acetyloleandomycin and 1,3-diacetyloleandomycin ( i n termediate metabolites) and 2,3-diacetyloleandomycin
(minor m e t a b o l i t e s ) .

TIME (hours)
Antibiotics Annual

Figure 6. Antibiotic activity vs. time curve in human


beings given 500 mg XLII (O), 500 mg XLIII (), and
250 mg XLIII (). Activity expressed in terms of XLII
base (168).

The o r a l b i o a v a i l a b i l i t y problems of erythromycin


(XLIV) are w e l l e s t a b l i s h e d (165-167)- E s t e r i f i c a t i o n
of XLIV at the 2 p o s i t i o n , ( i . e . , Hp = -COR- where
R 3 = a l k y l , a r y l , alkoxy or -(CH ) COOR4 where R4 =
a l k y l group) to give various e s t e r s was done w i t h the
expressed purpose of lowering the aqueous s o l u b i l i t y
of the erythromycin i n an attempt to decrease i t s
b i t t e r t a s t e . Many of these esters on o r a l dosing
gave s u p e r i o r or equivalent blood l e v e l s to e r y t h r o mycin base (176-181). The propionate e s t e r i n p a r t i c u l a r was considered to give superior l e v e l s of erythromycin when compared to erythromycin. There i s
some d i s c u s s i o n as to whether the conversion of the
2 -propionate ester to erythromycin i s complete i n
T

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

32

PRO-DRUGS

(XLIV): erythromycin
erythromycin

A, C
FT
B, R*
R

=
=
=
=

-OH,
-H
-H,
-H

v i v o . E s t e r i f i c a t i o n of erythromycin at the 2 , 4 " ,


and 1 1 p o s i t i o n s r e s u l t e d i n s i g n i f i c a n t increases i n
the l i p o p h i l i c i t y of the molecules. Whether these
esters r e v e r t t o the parent compound, maintain a c t i v i t y of t h e i r own, or impart any r e a l advantage over
erythromycin i t s e l f i s a c u r r e n t l y c o n t r o v e r s i a l t o p i c
( 1 8 0 , 1 8 2 ) . The a b i l i t y of a number of erythromycin
esters and s a l t s to mask the b i t t e r t a s t e of e r y t h r o mycin w i l l be discussed l a t e r .
The p o l a r a n t i b i o t i c , lincomycin (XLV) has been
e s t e r i f i e d at the 2 and 7 p o s i t i o n (XLVI) i n an
attempt to produce a t a s t e l e s s form of XLV s u i t a b l e
f o r p e d i a t r i c dosing ( 1 8 3 - 1 8 5 ) .
Obviously the b i o l o g i c a l p r o p e r t i e s of XLVI, such as absorption charact e r i s t i c s and regeneration of the parent compound,
a l s o need to be optimized. Good a c t i v i t y f o r 2 - a c y l
and 2-alkoxycarbonyl d e r i v a t i v e s were noted i n S.
aureus i n f e c t e d mice. The maximum a c t i v i t y was noted
for
through C^g e s t e r s with median chain length
e s t e r s having the b e t t e r a c t i v i t y ( 1 8 4 ) . The
F

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition


I 3

CH
H

33

(XLV) : R = R = -H
?
(XLVI) : 1
R and/or R = a c y l or
alkoxycarbonyl s u b s t i t u e n t
2,7-dicarbonate e s t e r s were found to be i n a c t i v e i n
v i t r o but had i n v i v o a c t i v i t y a f t e r S.C. i n j e c t i o n and
o r a l dosing i n S. aureus i n f e c t e d mice. A number of
the d e r i v a t i v e s (both a c y l and carbonate e s t e r s ) were
shown to achieve the o r i g i n a l o b j e c t i v e of the authors,
i . e . , a t a s t e l e s s lincomycin d e r i v a t i v e with i n v i v o
a c t i v i t y comparable to lincomycin base (185)
Mlek et a l . (l86) have attempted to increase del i v e r y of p o l a r a n t i b i o t i c s such as p e n i c i l l i n to p u l monary t i s s u e and lymph nodes by d e r i v a t i v e formation.
The authors argue that d e l i v e r y to the lymphatic system might be achieved i f drugs such as streptomycin,
neomycin and viomycin were a s s o c i a t e d as macromolecular
s a l t s with c a r b o x y l , s u l f o n y l or phosphoryl high molec u l a r weight polymers.
They conclude that these macromolecules, w i t h t h e i r c o l l o i d a l p r o p e r t i e s , have a high
a f f i n i t y f o r the lymphatic system. The authors prepared a number of macromolecular s a l t s of the various
a n t i b i o t i c s (the authors termed the r e s u l t a n t s a l t s
"antibiolymphins") i n c l u d i n g (a) p o l y a c r y l a t e s a l t s ,
(b) s a l t s w i t h s u l f o n y l and phosphorylated polysacchar i d e s , and (c) s a l t s w i t h n a t u r a l p o l y c a r b o x y l i c acids
of the polyuronic and polysaccharide s e r i e s . A f t e r
I.M., I.P., I.V., and i n t r a p l u r a l a d m i n i s t r a t i o n of the
macromolecular s a l t s of streptomycin, neomycin and v i o mycin, l e v e l s of the various drugs i n the lymphatic
system appeared to be higher than l e v e l s obtained from
a d m i n i s t r a t i o n of an equivalent dose of the nonderiva-

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

34

PRO-DRUGS

t i z e d drug.
The o r a l absorption of heavy metal c h e l a t i n g
agents such as ethylenediaminetetraacetic
a c i d (EDTA,
XLVII) and diethylenetriaminepentaacetic
a c i d (DTPA,
XLVIII) had been shown to be l e s s than k% of the t o t a l
dosage (187). The r e l a t i v e l y poor e f f e c t i v e n e s s of a
number of c h e l a t i n g agents t o promote r a d i o a c t i v e metal
m o b i l i z a t i o n has been a t t r i b u t e d t o t h e i r poor permeability characteristics. The chelating agents (2-hydroxyethylenediamine-N,N,N-triacetic acid (HEDTA, LXIX) and N,N'-bis(2-hyHOOCCH

^CH COOH

\cH CH N
HOOCCH
(XLIX)
.0
Hr-COOCCH

NCHCHN

H C OOCCH
5

(LI)

CH^COOH CH C00H
o

(LII)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

35

Overview and Definition


f

droxycyclohexyl)-ethylenediamine-N,N -diacetic a c i d
(DOC L) have been d e r i v a t i z e d to the l e s s polar compounds LI and L I I r e s p e c t i v e l y . A f t e r I.V. administrat i o n of L I , m o b i l i z a t i o n of various radiometals from
the l i v e r of Ce ^^ dosed cats was f a r s u p e r i o r to an
equivalent dose of XLIX. LI administered o r a l l y was as
e f f i c i e n t at radiometal m o b i l i z a t i o n as an equivalent
I.V. dose of L I . The increased o r a l b i o a v a i l a b i l i t y
and increased p e r m e a b i l i t y to various organs of LI and
LII were a t t r i b u t e d to the l e s s polar nature of the der i v a t i v e s and t h e i r ready conversion to the parent compound . The increased m o b i l i z a t i o n of radiometals by
LI was a t t r i b u t e d to the formed XLIX because LI was
shown to have no heav i o c h e l a t i n capacit (188)
5

Opthalmic absorptio o epinephrine.


e highly
p o l a r adrenergic agent, epinephrine ( I X ) , i s u s e f u l i n
the treatment of glaucoma. As demonstrated e a r l i e r ,
l e s s than 1% of IX i s present i n i t s n e u t r a l form at
p h y s i o l o g i c a l pH. A c y l a t i o n of the phenolic hydroxy
groups to give the d i p i v a l y l d e r i v a t i v e ( L I I I ) was
found to increase the therapeutic e f f e c t i v e n e s s of IX
by a f a c t o r of approximately 100 (to be discussed f u r ther by Dr. McClure). Even though the f r a c t i o n of

CHCH NH(CH )

(CH ) CC0
Q

(LIII)
n e u t r a l molecule present at p h y s i o l o g i c a l pH should not
be g r e a t l y a f f e c t e d by the a c y l a t i o n , the l i p i d s o l u b i l i t y of L I I I i s f a r superior to i t s parent compound,
IX. Since corneal absorption i n v o l v e s transport
through a l i p o i d a l b a r r i e r , the greater l i p o p h i l i c i t y
of L I I I may account f o r i t s superior therapeutic e f f e c t i v e n e s s . The use of the d i p i v a l y l e s t e r was necessary
not only to increase the l i p o p h i l i c i t y of the compound
but a l s o to help guarantee adequate aqueous s t a b i l i t y .
Percutaneous absorption of p o l a r drug e n t i t i e s .
The absorption of drugs i n t o and through the s k i n i s an
area of study which has achieved wide coverage but
which i s not, as y e t , f u l l y understood. I t i s generall y accepted that only n e u t r a l , r e l a t i v e l y l i p o i d a l drug

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

36

PRO-DRUGS

molecules can be absorbed percutaneously (19*0 The


work of Imai et_ a l . ( 1 8 9 ) on the percutaneous absorpt i o n of the p o l a r v i t a m i n , a s c o r b i c a c i d (LIV) and i t s
h i g h l y p o l a r 3-phosphoryl e s t e r (LV) tends to contrad i c t t h i s g e n e r a l i z a t i o n . The relevancy of t h i s work

1>=0

H-

-OH

H-

-OH
H

(LIV);
(LV):

R = -H
R = -P0 H
3

and the true e f f e c t i v e n e s s of percutaneous absorption


of LIV from LV i s not known at t h i s time.
The s u l f a drug mafenide (LVI) used i n burn therapy
was r e l a t i v e l y i n e f f e c t i v e when a p p l i e d percutaneously
as i t s hydrochloride s a l t ( L V I I ) . However, a p p l i c a t i o n
of the acetate s a l t of mafenide ( L V I I I ) was found to be
very e f f e c t i v e i n burn treatment (12-14). Mafenide i s
marketed as Sulfamylon ( L V I I I i n a watermiscible
cream formulation) and Sulfamylon hydrochloride s o l u t i o n (LVII as a 5% aqueous s o l u t i o n ) . The i n e f f e c t i v e ness of the 5% aqueous s o l u t i o n of LVII i n the t r e a t ment of burns has been discussed (190). The d i f f e r e n c e
i n a c t i v i t y between LVII and L V I I I i s an i n t e r e s t i n g
problem. The pKa of mafenide i s 8.52 at 21C (191).
A 5% aqueous s o l u t i o n of LVII would be expected to have
a pH of approximately 4.5. The pH of an aqueous f i l m
of L V I I I , regardless of c o n c e n t r a t i o n , would be expected t o have a pH of approximately 6.6. Therefore, the
f r a c t i o n of mafenide present i n i t s n e u t r a l and presumably absorbable form, i n a 5% s o l u t i o n of LVII i s approximately 0.01$ whereas t h e / f r a c t i o n present i n an
aqueous s o l u t i o n of L V I I I i s approximately 1%. A model
f o r the absorption of mafenide i n t o the s k i n i s shown
x

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

37

Overview and Definition

( L V I I ) ; X = CI
(LVIII); X =

CH3COO

i n Scheme V I . Whethe
depend on whether i s a weak or strong base or wheth
er LVII or L V I I I i s i n buffered s o l u t i o n s . Since the
acetate anion i s a much stronger base than the c h l o r i d e
anion, the e q u i l i b r i u m i s forced t o the r i g h t f a v o r i n g
the formation of LVI. L V I I I i s considered a pro-drug
of LVI, yet i t i s simply a s a l t of LVI and regeneration
of LVI from L V I I I i n v o l v e s simple d i s s o c i a t i o n .
S t e r o i d s are an area where the pro-drug approach
has been apparently s u c c e s s f u l l y u t i l i z e d i n attempts
to promote t o p i c a l or percutaneous absorption. Ste
r o i d s such as triamcinolone ( L I X ) , f l u o c i n o l o n e (LX)
and f l u c l o r o l o n e have been d e r i v a t i z e d t o t h e i r acetonide d e r i v a t i v e s (192), triamcinolone acetonide

Scheme VI

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

38

PRO-DRUGS

(LXa), f l u o c i n o l o n e acetonide (LXb), f l u o c i n o l o n e acetonide-21-acetate (LXc), f l u c l o r o l o n e acetonide and


desonide. Ester d e r i v a t i v e s of various other s t e r o i d s
such as the v a l e r a t e ester of betamethasone, the p r o p i onate ester of c l o b e t a s o l , the p i v a l a t e and hexanoate
esters of f l u c o c o r t o l o n e , the acetate ester of hydroc o r t i s o n e , the p i v a l a t e e s t e r of flumethasone, the d i propionate ester of beclomethasone, and the acetate
ester of methylprenisolene are examples of a few of
the c o r t i c o s t e r o i d esters i n use (192,193)
T o p i c a l c o r t i c o s t e r o i d s are used i n the treatment
of inflammatory, a l l e r g i c and p r u r i t i c s k i n c o n d i t i o n s .
Whether any of the pro-drug d e r i v a t i v e s exert any a n t i inflammatory a c t i v i t
f thei
whethe the
quire conversion t
i t y can probably be argued.
Poulsen (194) has r e c e n t l y discussed formulation
f a c t o r s a f f e c t i n g the percutaneous d e l i v e r y of drugs.
As pointed out by Poulsen, there are many f a c t o r s a f f e c t i n g the dermal d e l i v e r y of a drug. I t appears that
f o r s o l u t i o n s , g e l s , creams, e t c . , the d i f f u s i o n of
drug across the s k i n b a r r i e r i s rate l i m i t i n g . I n such
cases, the p h y s i c a l and chemical properties of the drugto be d e l i v e r e d and the v e h i c l e which contains the drug
are of paramount importance. I n p a r t i c u l a r , the a c t i v i t y of the drug i n the v e h i c l e or the e f f e c t i v e concent r a t i o n of the drug i n the v e h i c l e determines, "the
d r i v i n g force f o r d i f f u s i o n from the v e h i c l e (194).
The d i f f u s i o n c o e f f i c i e n t , the a c t i v i t y of a drug
i n a v e h i c l e , and the p a r t i t i o n c o e f f i c i e n t of a drug
between the stratum corneum and the v e h i c l e are a l l a f fected by the p h y s i c a l properties of a drug. Acetonide
formation of the dihydroxy groups i n some s t e r o i d s
(Scheme VII) or e s t e r i f i c a t i o n of hydroxy groups i n
other s t e r o i d s generally r e s u l t s i n an a p r i o r i i n crease i n p a r t i t i o n c o e f f i c i e n t between the stratum
corneum and a h y d r o p h i l i c v e h i c l e . This causes an i n crease i n the d i f f u s i o n constant across human stratum
corneum and lowers the s o l u b i l i t y of the s t e r o i d i n the
hydrophilic vehicle.
Poulsen points out that the q u a n t i t a t i v e d i f f e r ences i n anti-inflammatory a c t i v i t y between various der i v a t i v e s of an agent i s d i f f i c u l t t o judge because of
v e h i c l e e f f e c t s (194). For example, i n the comparison
of the anti-inflammatory a c t i v i t y of LXb t o LXc (administered as a carboxypolymethylene g e l containing
various r a t i o s of propylene glycol/water as the s o l v e n t ) , LXb was found t o be more a c t i v e than LXc (human
v a s o c o n s t r i c t o r assay) when the percentage of propylene
g l y c o l i n the g e l was <55% but was found t o be l e s s
11

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

39

(LXb)
Scheme V I I
a c t i v e when the percentage of propylene g l y c o l i n the
v e h i c l e was >55$ These d i f f e r e n c e s were a t t r i b u t e d t o
d i f f e r e n c e s i n s o l u b i l i t i e s of LXb and LXc i n the ve
h i c l e as a f u n c t i o n of propylene g l y c o l concentration.
LXb (0.025$) was completely soluble i n the v e h i c l e when
the percentage of propylene g l y c o l was >20-40$ while
LXc (0.025$) d i d not d i s s o l v e completely i n the v e h i c l e
u n t i l the percentage of propylene g l y c o l reached ap
proximately 80$. The importance of v e h i c l e composi
t i o n , see Poulsen (194) f o r a complete d i s c u s s i o n of
t h i s problem, i n the anti-inflammatory a c t i v i t y of s t e
r o i d s and t h e i r d e r i v a t i v e s was r e c e n t l y re-emphasized
by the study of Barry and Woodford (192).
M a i s t r e l l o et a l . ( 1 9 5 ) have r e c e n t l y attempted t o
quantitate the t o p i c a l anti-inflammatory e f f e c t s of
various s t e r o i d a l agents administered as s o l u t i o n s i n
2-(2-ethoxyethoxy)ethanol.
The greater a c t i v i t y of LXa
r e l a t i v e to LIX was r e a d i l y apparent.
The pro-drug approach has been s u c c e s s f u l i n im
proving the anti-inflammatory c l i n i c a l e f f i c a c y of per
cutaneously administered s t e r o i d s as can be seen by the
large number of s t e r o i d s c u r r e n t l y a v a i l a b l e as e i t h e r
esters or acetonide d e r i v a t i v e s ( r e l a t i v e to nonderivat i z e d s t e r o i d s ) . However, the r o l e of the v e h i c l e i n
the success of these products has only r e c e n t l y begun
to be understood and f u l l y appreciated (194) .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

40

PRO-DRUGS

To Increase the Aqueous S o l u b i l i t y of Drugs to


Help F a c i l i t a t e Oral Absorption.
Rather s u r p r i s i n g l y ,
there are few examples of the pro-drug approach being
used to increase the aqueous s o l u b i l i t y of poorly water
s o l u b l e drugs i n an e f f o r t to increase the o r a l absorp
t i v i t y of the drugs. The c l a s s i c a l examples of im
proved water s o l u b i l i t y of poorly water s o l u b l e drugs
are those f o r which an I.V. i n j e c t a b l e form of the drug
was d e s i r e d . The apparent l a c k of examples may be due
to the f a c t that improvement i n the o r a l absorption of
a drug can o f t e n be e f f e c t e d by formulation techniques.
The o r a l b i o a v a i l a b i l i t y of 5,5-diphenylhydantoin,
n i t r o f u r a n t o i n , g r i s e o f u l v i n , d i g o x i n , prednisolone,
etc., has been s u c c e s s f u l l y improved by formulation
techniques.
The poor and e r r a t i c o r a b i o a v a i l a b i l i t y o d i
goxin due t o i t s low water s o l u b i l i t y and formulation
v a r i a b l e s has been w e l l e s t a b l i s h e d (196-200).
Higuchi
and Ikeda (201) have r e c e n t l y demonstrated that the
complex between hydroquinone and d i g o x i n (2 d i g o x i n :
3 hydroquinone) had a much higher d i s s o l u t i o n r a t e than
digoxin i t s e l f .
Any c r y s t a l l i n e m a t e r i a l such as d i g o x i n w i l l have
a d i s s o l u t i o n r a t e h i g h l y dependent on i t s aqueous
s o l u b i l i t y as w e l l as other v a r i a b l e s defined i n the
Noyes-Whitney equation (202). The energetics of the
d i s s o l u t i o n process are determined by the breakdown of
i n t e r m o l e c u l a r forces i n the c r y s t a l l a t t i c e and the
s o l v e n t , both r e q u i r i n g energy, r e l a t i v e to the r e l e a s e
of s o l v a t i o n energy due to s o l u t e - s o l v e n t i n t e r a c t i o n s .
The high m e l t i n g point ( 2 6 5 with decomposition) of
d i g o x i n s t r o n g l y suggested that c r y s t a l l a t t i c e energy
played an important r o l e i n the poor aqueous s o l u b i l i t y
of d i g o x i n . The complexation of d i g o x i n w i t h a polyhydroxy compound such as hydroquinone might d i s r u p t
t h i s t i g h t c r y s t a l l a t t i c e to form a complex with a
s u p e r i o r aqueous s o l u b i l i t y . On d i s s o l u t i o n , d i s s o c i a
t i o n of the complex would r a p i d l y release d i g o x i n .
" I n t r i n s i c a l l y more r a p i d l y d i s s o l v i n g forms of d i g o x i n
would provide greater assurance of more reproducible
and more b i o a v a i l a b l e d i g o x i n products" (201).
Use of d i g o x i n d e r i v a t i v e s such as i t s 4 ' - m e t h y l
d e r i v a t i v e (203-205) and a c y l a t e d d e r i v a t i v e s such as
acetyldigoxin- 26,207), a c e t y l d i g o x i n - 3 (203*208211) have been promoted. The acylated d e r i v a t i v e s of
g i t o x i n (212-214), as w e l l as c y c l i c a c e t a l s and acet a l s of d i g i t o x i n , d i g o x i n , and quabain have been of
i n t e r e s t (215). Although 4 ' - a c e t y l d i g o x i n ( L X I I ) has
been shown to regenerate d i g o x i n ( L X I ) , 4 ' ' m e t h y l d i goxin ( L X I I I ) was not thought to r e v e r t to d i g o x i n .
ft

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

41

0
II

OH H
However, the studies of Rietbock e_t a l . (204) confirm
that demethylation of L X I I I does occur. Whether any of
these d e r i v a t i v e s , monoalkyl ( 2 0 3 - 2 0 5 ) monoacyl ( 2 0 6 214), or p o l y a c y l ( 2 1 6 ) , have any r e a l advantage over
d i g o x i n i t s e l f i s debatable. White and G r i s v o l d ( 2 1 7 )
claim good o r a l absorption p r o p e r t i e s f o r LXII i n cats
and LXII i s marketed as Acylanid (Sandoz) i n the
United S t a t e s .
Recently, Hussain and R y t t i n g ( 2 1 8 ) argued that
a l l o p u r i n o l (LXIV) owed i t s low water s o l u b i l i t y of
0 . 7 8 mg/ml to strong i n t e r m o l e c u l a r hydrogen bonding i n
i t s c r y s t a l l a t t i c e . A melting point of 3 6 5 f o r LXIV
seemed to confirm t h i s assumption. D i s r u p t i o n of the
c r y s t a l l a t t i c e by t r a n s i e n t pro-drug formation was
suggested as a means of i n c r e a s i n g the aqueous s o l u b i l i t y of a l l o p u r i n o l . The authors synthesized 1e t h o x y e t h y l - 4 - a l l o p u r i n y l ether (LXV) and 2 - t e t r a h y d r o p y r a n y l - 4 - a l l o p u r i n y l ether (LXVI) and demonstrated
the improved d i s s o l u t i o n r a t e from a constant surface
area p e l l e t of LXV and LXVI when compared to LXIV.
LXV and LXVI were shown to regenerate LXIV under a c i d i c
0

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

42

PRO-DRUGS

c o n d i t i o n s . Unfortunately the authors d i d not attempt


to confirm the p r e d i c t e d improved b i o a v a i l a b i l i t y of
LXIV from i t s pro-drug d e r i v a t i v e s by i n v i v o s t u d i e s .
However, a u s e f u l conclusion from the work i s that improved aqueous s o l u b i l i t y of an agent need not necess a r i l y r e q u i r e d e r i v a t i z a t i o n to a water soluble s a l t

(LXIV):

R = -H

(LXV):

R = -CH (CH )-OC H


2

(LXVI):
form of the drug. I f the determining f a c t o r t o poor
aqueous s o l u b i l i t y i s the strength of the c r y s t a l l a t t i c e of the drug, and t h i s c r y s t a l energy i s not s u f f i c i e n t l y r e l i e v e d by s o l v a t i o n energy on d i s s o l u t i o n ,
then d i s r u p t i o n of the c r y s t a l l a t t i c e by pro-drug f o r mation may provide a s i g n i f i c a n t increase i n aqueous as
w e l l as l i p i d s o l u b i l i t y .
The strong c r y s t a l l a t t i c e energies of the hydant o i n s , 5*5-diphenylhydantoin and n i t r o f u r a n t o i n , and
various pro-drug forms of these drugs w i l l be discussed
l a t e r by S t e l l a et a l .
The a n t i f u n g a l agent, g r i s e o f u l v i n (LXVII), has
been shown to be poorly absorbed a f t e r o r a l administrat i o n t o man as w e l l as animals (219-225) The study by
Bates et a l . (226) has shown that the absorption of
g r i s e o f u l v i n can be g r e a t l y enhanced by the concomital
a d m i n i s t r a t i o n of f a t s . I t appears that the incomplete
b i o a v a i l a b i l i t y of LXVII i s a f u n c t i o n of i t s low water
s o l u b i l i t y . This can be traced t o i t s high l i p o p h i l i c i t y , a contrast to the c r y s t a l l a t t i c e s t r u c t u r e problems associated with the e a r l i e r examples. F i s c h e r and
Riegelman (227) attempted t o increase the aqueous s o l u b i l i t y of LXVII by pro-drug formation. The d e r i v a t i v e s

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

43

studied were g r i s e o f u l v i n - 4 ' - a l c o h o l ( L X V I I I ) , g r i s e o f u l v i n - ^ -oxime (LXIX), griseofulvin-4'-carboxymethoxime (LXX) and griseofulvin-4'-hemisuccinate (LXXI).
In a l l d i s c u s s i o n s thus f a r , most of the d e r i v a t i v e s
were e s t e r s of the parent compound where regeneration
of the parent compound was p o s s i b l e by the known presence and abundance of esterases i n the body. The s a l t
and complex pro-drugs reverted to the parent compounds
by d i s s o c i a t i o n (a nonenzymatic process). In the case
of g r i s e o f u l v i n pro-drugs, the conversion of LXVIII to

CI

(LXVII):
(LXVIII):
(LXIX):
(LXX):
(LXXI):

CH

R
R
R
R
R

=
=
=
=
=

=0
-OH
=N0H
=N0CH C00H
-OCOCHpCHpCOOH
p

LXVII r e q u i r e s an o x i d a t i v e metabolism. On I.V. dosing


to a r a b b i t , the disappearance of LXVIII from the p l a s ma of the r a b b i t had a h a l f - l i f e of 28 minutes whereas
the formed g r i s e o f u l v i n had a h a l f - l i f e of 70 minutes.
LXXI i s converted to LXVII by esterase h y d r o l y s i s of
the e s t e r f u n c t i o n followed by o x i d a t i o n of the formed
LXVIII. Oximes have a l s o been shown to be enzymaticall y converted to the corresponding ketone (228).
Although each of the LXVII pro-drugs had s u p e r i o r
aqueous s o l u b i l i t i e s when compared to LXVII (227)*
none of the d e r i v a t i v e s on o r a l dosing showed any sup e r i o r i t y over LXVII i t s e l f . This was p o s t u l a t e d to be
due to e i t h e r incomplete conversion of the d e r i v a t i v e s
to LXVII, concurrent metabolism to i n a c t i v e metabol i t e s , or e l i m i n a t i o n from the body before conversion
to LXVII was complete.
To Help S t a b i l i z e Drugs Against Metabolism and/or
H y d r o l y s i s During Oral Absorption. Many drugs are extremely a c t i v e i f administered p a r e n t e r a l l y but s u f f e r
from the problem of incomplete absorption on o r a l

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

44

PRO-DRUGS

dosing. This incomplete absorption, as has already


been noted, can r e s u l t from the drug being too p o l a r or
poorly water s o l u b l e . A t h i r d p o s s i b l e reason f o r i n complete absorption i s that the drug, i f administered
o r a l l y , may be r a p i d l y metabolized by enzymes secreted
i n t o the GI t r a c t , by b a c t e r i a i n the GI t r a c t , by enzymes encountered while passing through the GI mucosa,
and/or by the l i v e r i n i t s i n i t i a l t r a n s i t through the
l i v e r before ever reaching the general c i r c u l a t i o n .
Examples of drugs poorly absorbed due to one or a comb i n a t i o n of these processes have already been d i s cussed, e.g., L-Dopa. This o v e r a l l process of incomp l e t e systematic a v a i l a b i l i t y on o r a l absorption due to
metabolism during the absorption process has been
termed the " f i r s t pass
" f i r s t pass" e f f e c t
originally
g
only l i v e r e x t r a c t i o n (30).
I t has become l e s s w e l l
defined and i s used g e n e r i c a l l y to describe dose dependent b i o a v a i l a b i l i t y due to metabolism of o r a l l y adm i n i s t e r e d drugs during the absorption process.
Magee et a l . (229) have r e c e n t l y studied the i n s i t u
absorption of various prostaglandins from the small i n t e s t i n e of r a t s . Although disappearance from the r a t
lumen was f a i r l y r a p i d f o r a l l the p r o s t a g l a n d i n s , very
l i t t l e of the dose a c t u a l l y appeared i n the blood
stream and only a small f r a c t i o n of t h i s " e f f e c t i v e l y "
absorbed dose was i n t a c t p r o s t a g l a n d i n . The p r o s t a g l a n d i n , 15-methyl P a (LXXII) and i t s methyl e s t e r
(LXXIII) were s t u d i e d . LXXII had an apparent absorpt i o n h a l f - l i f e of 60-70 minutes w i t h approximately 2$
of the dose reaching general c i r c u l a t i o n but of that
only 0.8$ was i n t a c t LXXII. When LXXIII was administered up to 7$ of the dose reached general c i r c u l a t i o n , and 1.8$ of the dose was present i n serum as i n 2

OH

(LXXII): R = -H
(LXIII):

R =

-CH

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

45

t a c t LXXII w i t h l i t t l e or no detectable LXXIII. This


i s a c l e a r example of an agent, n o t o r i o u s l y suscept i b l e to metabolism, that was chemically modified to
p a r t i a l l y overcome the " f i r s t pass" e f f e c t .
That e s t e r i f i c a t i o n of prostaglandins i n h i b i t s
the metabolism of prostaglandins was c l e a r l y demons t r a t e d by the greater a c t i v i t y of the methyl e s t e r of
p r o s t a g l a n d i n 15-methyl
over the parent non-methyl
e s t e r p r o s t a g l a n d i n . The greater potency i s a t t r i buted to a lengthening of the metabolic h a l f - l i f e (230)
of the e s t e r form r e l a t i v e to the parent a c i d p r o s t a glandin.
Another group of compounds which undergoes considerable metabolism during o r a l absorption are s t e r o i d s
The b i o a v a i l a b i l i t
unknown because of a n a l y t i c a
i t y and i n t e r f e r e n c e from n a t u r a l s t e r o i d a l hormones
i n analyzing f o r absorbed i n t a c t s t e r o i d . Another
c o n t r i b u t i o n to t h i s unknown b i o a v a i l a b i l i t y i s that
many of the s t e r o i d s are probably metabolized to act i v e agents.
Schedl et a l . (231) showed that the r a t e of abs o r p t i o n of various s t e r o i d s using the i n s i t u r a t i n t e s t i n a l loop experiment was r a p i d and i n v e r s e l y prop o r t i o n a l to the number of hydroxy or p o l a r groups
present i n the molecule.
This observation was confirmed when many of the s t e r o i d s were a c e t y l a t e d and
the absorption r a t e of the a c e t y l a t e d versus the nona c e t y l a t e d s t e r o i d s compared. Although many of the
s t e r o i d s were found to be r a p i d l y absorbed, the weak
o r a l potency of a number of the agents, e.g., t e s t o s terone, progesterone and desoxycorticosterone, which
were apparently w e l l absorbed, the authors s t a t e that
the "pharmacologic a c t i v i t y of a s t e r o i d by the o r a l
route i s independent of i t s absorption r a t e . Blood
l e v e l s of s t e r o i d s f o l l o w i n g o r a l a d m i n i s t r a t i o n are
more a f u n c t i o n of metabolic d i s p o s i t i o n than of abs o r p t i o n r a t e " (231). A c e t y l a t i o n may not only help
increase the absorption r a t e of these s t e r o i d s but may
a l s o provide a degree of metabolic p r o t e c t i o n .
Tanabe et a l . (232) and P r i e d et a l . (233) have
shown that 17ot,21-acetonides of various c o r t i c o s t e r o i d s have higher o r a l a c t i v i t y than t h e i r parent s t e r o i d s on o r a l dosing. The study of P r i e d et a l . (233)
demonstrated the higher a c t i v i t y of l6a,17a-acetonide
of triamcinolone (LXXIV) over the parent s t e r o i d
(LXXV). Gardi et a l . (234) a l s o observed that p r e d n i solone acetonide (LXXVI) when administered o r a l l y and
prednisolone cyclopentylidenedioxy (LXXVII) when app l i e d l o c a l l y had greater a c t i v i t y than the parent

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

46

PRO-DRUGS

CH 0H
I
c=o
o
2

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

ilu
1.

STELLA

Overview and Definition

ary

a m i c a l Society

47

prednisolone. The a n t i f e r t i l i t y , anti-inflammatory


compound 9ct,ll$ , 2 1 - t r i c h l o r o - l 6 a , 1 7 a - ( i s o p r o p y l i d e n e d i o x y ) - l , 4 , 6 - p r e g n a t r i e n e - 3 5 2 0 - d i o n e , an acetonide der i v a t i v e , was a l s o shown to be o r a l l y a c t i v e (235).
The s t e r o i d desoxycorticosterone (LXXVIII) i s
very unstable and d i f f i c u l t to handle but i t s acetate
e s t e r has been used p a r e n t e r a l l y i n the treatment of
Addison's disease. LXXVIII as i t s acetate e s t e r has
been found to be destroyed a f t e r o r a l a d m i n i s t r a t i o n
but when given s u b l i n g u a l l y was found to be u s e f u l i n
the treatment of Addison's disease (236).
Gardi et a l . (237) have r e c e n t l y demonstrated the
high o r a l a c t i v i t y of l , 3 , 5 ( 1 0 ) - e s t r a t r i e n - 1 7 6 - y l enol
ethers and a c e t a l s a
classe
f orall
d
t e r a l l y a c t i v e estrogeni
i s a t t r i b u t e d to th
regenerate
Moreover the authors s t a t e that "the ether linkage
should be s t a b l e enough to survive the a c i d i c g a s t r i c
medium and s u i t a b l y delay the hepatic i n a c t i v a t i o n
after oral administration."
That catecholamines show poor absorption through
l i p o i d a l membranes was e s t a b l i s h e d e a r l i e r i n t h i s
manuscript.
" F i r s t pass" metabolism, p r i m a r i l y due to
conjugation ( g l u c u r o n i d a t i o n and/or s u l f a t i o n ) , was
also mentioned as a primary means of l i m i t i n g the systemic a v a i l a b i l i t y of catecholamines and other phenol i c compounds. The n a r c o t i c antagonist naloxone
(LXXIX) has been shown to have low potency after oral
administration (238-241). The short duration of action of
LXXIX a f t e r p a r e n t e r a l dosing due to r a p i d metabolism
suggests that naloxone might be r a p i d l y metabolized by
the l i v e r and/or g a s t r i c mucosa on o r a l dosing, i . e . ,
the poor o r a l a c t i v i t y r e s u l t s from a " f i r s t pass" e f f e c t . Linder and Fishman (240) synthesized a s e r i e s
of s u l f a t e and acetate e s t e r s of LXXIX and t e s t e d
t h e i r n a r c o t i c antagonist a c t i v i t y a f t e r o r a l and pare n t e r a l dosing i n r a t s . The 3-acetyl d e r i v a t i v e
(LXXX), 14-acetyl d e r i v a t i v e (LXXXI) and 3,14-diacetyl
d e r i v a t i v e (LXXXII) a l l showed good o r a l a c t i v i t y i n
the morphine challenge t e s t whereas LXXIX administered
o r a l l y was r e l a t i v e l y i n e f f e c t i v e . A f t e r I.V. adminis t r a t i o n , both LXXX and LXXXI were more potent than
LXXIX whereas LXXXII was s l i g h t l y l e s s potent than
LXXIX. These r e s u l t s suggest that the a c e t y l a t i o n of
the 3 and/or 14 hydroxy groups of LXXIX blocked (or
p a r t i a l l y blocked) the " f i r s t pass" metabolism of
LXXIX (probably s u l f a t i o n and/or g l u c u r o n i d a t i o n ) by
p r o t e c t i n g the hydroxy groups.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

48

PRO-DRUGS

(LXXIX):
(LXXX);
(LXXXI)
(LXXXII)

Rt
K: = -COCHo, R^ = -H
R^ = -H,
R = -COCH.
R = R = -COCH^
2

Way and Adler (242), i n t h e i r review of morphine


(LXXXIII), morphine m e t a b o l i t e s , and other n a r c o t i c
a n a l g e t i c s , s t a t e that the poor o r a l a c t i v i t y of morphine r e l a t i v e t o i t s p a r e n t e r a l a c t i v i t y may be due
to the poor o r a l absorption of morphine. The e s s e n t i a l l y negative pharmacological a c t i v i t y o f o r a l l y adm i n i s t e r e d morphine i n man can be i n t e r p r e t e d as e i t h e r
poor i n t r i n s i c absorption and/or " f i r s t pass" metabol i s m . Heroin or 3 6-diacetylmorphine (LXXXIV) e x h i b i t s
3

CH

N-

(LXXXIII):
(LXXXIV):

R = -H
R = -COCH

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

49

considerable i f not somewhat i r r e g u l a r a c t i v i t y i n man


a f t e r o r a l dosing which i s again c o n s i s t e n t with the
b l o c k i n g (or p a r t i a l blocking) of " f i r s t pass" metabo
l i s m . The r a p i d d e a c e t y l a t i o n of LXXXIV i n a l l b i o l o g
i c a l t i s s u e s , i n c l u d i n g the CNS, to LXXXIII and i t s mo
noesters i s w e l l e s t a b l i s h e d (242). Kupchan et^ a l .
(243) synthesized l a b i l e ether d e r i v a t i v e s of morphine
and phenazocine and showed t h e i r a c t i v i t y to be l e s s
than those of the parent compounds. Papers by Y o s h i mura et a l . (244), Oguri et a l . (245) and Mori et a l .
(246) have confirmed the presence of glucuronide and
s u l f a t e metabolites of morphine. Their r e s u l t s sur
p r i s i n g l y show that the 6-glucuronide and 6 - s u l f a t e me
t a b o l i t e s when i n j e c t e d S.C
i mic (246) giv highe
potency and l a r g e r d u r a t i o
pared to a comparabl
morphine
pointe
by the authors, both the 6-glucuronide and 6 - s u l f a t e
are minor metabolites of morphine.
In a s e r i e s of papers by F i e l d s et a l . (247-250),
various l a t e n t i a t e d forms of t h i o l s , amino-thiols and
2-acetamidoethanethiol were attempted to help reduce
the t o x i c i t y and/or improve a c t i v i t y , i . e . , increase
a v a i l a b i l i t y , but w i t h marginal success.
Similarly,
H a r t l e s et a l . (251) and Siuda et a l . (252) synthesized
and t e s t e d f o r a c t i v i t y a number of a c y l and carbamate
d e r i v a t i v e s of mafenide (LVI). Their o b j e c t i v e was to
f i n d o r a l l y a c t i v e forms of LVI. LVI was shown to un
dergo rapid^metabolism on o r a l absorption and i t was
f e l t that - a c y l and N^-alkoxycarbonyl d e r i v a t i v e s
might prevent the r a p i d metabolism so r e s u l t i n g i n ade
quate blood l e v e l s . Regeneration of LVI from i t s N^a c y l and N^-alkoxycarbonyl d e r i v a t i v e s was f e l t to be
too slow and inadequate to produce reasonable blood
l e v e l s of LVI.
Recently the t u b e r c u l o s t a t i c agent, p-aminosalic y l i c a c i d (LXXXV), has been shown to undergo " f i r s t
pass" metabolism due to N - a c e t y l a t i o n (253) . In e f
f o r t s to o b t a i n t a s t e l e s s forms of LXXXV, various chem
i c a l and formulation m o d i f i c a t i o n s of LXXXV have been
attempted. Most of these m o d i f i c a t i o n s r e s u l t i n a
slower release form of LXXXV which i s then more r e a d i l y
and e f f i c i e n t l y metabolized.
However, an i n t e r e s t i n g
pro-drug of LXXXV, the calcium 4-benzamidosalicylate
(LXXXVI), which i s q u i t e water i n s o l u b l e has been shown
to be as c l i n i c a l l y e f f e c t i v e on a molar basis as LXXXV
and yet t a s t e l e s s (254,255). This appears p a r a d o x i a l
i n that most slow release forms of LXXXV provide incom
p l e t e b i o a v a i l a b i l i t y due to the " f i r s t pass" e f f e c t .
I t appears that LXXXVI may provide a poorly s o l u b l e ,
t a s t e l e s s form of LXXXV while circumventing " f i r s t

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

50

PRO-DRUGS

pass" a c e t y l a t i o n (the aromatic group already being


acylated) and once i n the body regenerates LXXXV. The
r a t e of d e a c y l a t i o n of N-acyl d e r i v a t i v e s of drugs
other than formyl d e r i v a t i v e s and benzoyl d e r i v a t i v e s
i s poor. Chiou ( 2 5 6 ) has shown that deformylation o f
4,4 -diformamidodiphenylsulfone (DFD, LXXXVII) t o the
a n t i m a l a r i a l agent, 4,4-diaminodiphenylsulfone (DDS,
LXXXVIII) by kynurenine formamidase of mammalian l i v e r s
does occur (see scheme V I I I ) .
f

Ca(H 0)

J 2

(LXXXVI)

Lj yu \
(LXXXVII)
KYNURENINE
FORMAMIDASE

(LXXXVIII)

Scheme V I I I

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

51

Overview and Definition

The Use of Pro-Drugs to E f f e c t Sustained or


Release

Prolonged

Sustained r e l e a s e has u s u a l l y been e f f e c t e d i n


pharmacy by the use of various dosage form changes such
as coated slow r e l e a s e beads and granules, m u l t i p l e
l a y e r t a b l e t s , and other formulation techniques (257)
Many of the examples of pro-drugs causing sustained r e lease incorporate a formulation technique combined w i t h
chemical m o d i f i c a t i o n ideas.
Stempel (257) has described the advantages of prolonged or sustained r e l e a s e products:
(a) reduces the
number and frequency of doses needed to be administ e r e d ; (b) e l i m i n a t e th "peak"
d "valley" effect
noted w i t h conventiona
o f t e n reduces the t o t a
g
the d e s i r e d pharmacological a c t i v i t y ; (d) e l i m i n a t e s
the problem of nighttime a d m i n i s t r a t i o n of drugs; (e)
helps minimize the problem of p a t i e n t noncompliance by
decreasing the number of times a p a t i e n t must remember
to take t h e i r medication; ( f ) reduces the incidence of
peak blood l e v e l s r i s i n g above the t o x i c blood l e v e l s ;
(g) reduces the incidence of GI side e f f e c t s . To maint a i n an e f f e c t i v e blood l e v e l of the very short h a l f l i v e d drug cytosine arabinoside (LXXXIX), long term
I.V. i n f u s i o n s of considerable inconvenience to the
p a t i e n t and nursing s t a f f are needed. 5 - A c y l d e r i v a t i v e s of LXXXIX (an immunosuppressive, a n t i v i r a l and
c y t o t o x i c agent) as w e l l as 2 and 3 esters have been
synthesized and t e s t e d (258-262). When the s l i g h t l y
water s o l u b l e 5 - a c y l d e r i v a t i v e s (XC) were administered I.P. to mice as suspensions the pro-drugs d i s solved s l o w l y , g r a d u a l l y r e l e a s i n g XC to the c i r c u l a t i o n where the d e r i v a t i v e s were enzymatically cleaved
to LXXXIX a l l o w i n g i t to exert i t s pharmacological act i v i t y . Gray et a l . (258) demonstrated a q u a l i t a t i v e
c o r r e l a t i o n between a c t i v i t y of the XC d e r i v a t i v e s and
decreasing aqueous s o l u b i l i t y suggesting that the slow
d i s s o l u t i o n of the suspension of the 5 - e s t e r s was important. The s l i g h t l y s o l u b l e e s t e r s such as the 5 p a l m i t a t e , 5'-starate, 5 -benzoate and 5 -adamantoate
were p a r t i c u l a r l y impressive. The s l i g h t l y water s o l uble but s t e r i c a l l y hindered e s t e r , 2,4,6-trimethylbenzoate and the sulfonate e s t e r , 2 , 4 , 6 - t r i i s o p r o p y l benzene sulfonate were l e s s e f f e c t i v e than the parent
compound, presumably because of t h e i r slower enzymatic
cleavage to the parent compound.
Probably the area i n which the greatest e f f o r t has
been made to e f f e c t sustained or prolonged r e l e a s e act i v i t y v i a the pro-drug approach has been i n the area
1

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

52

PRO-DRUGS

OH

(LXXXIX):
(XC);

R = -H
R = a c y l substituent

of s t e r o i d therapy.
Recently Tanaka et a l . ( 2 6 3 ) have
attempted to q u a n t i t a t e f a c t o r s which a f f e c t the prol o n g a t i o n of a c t i v i t y of drug i n j e c t e d I.M. i n o i l y
s o l u t i o n s . James et a l . ( 2 6 4 ) showed that f o r a s e r i e s
of testosterone (XCI) esters the b i o l o g i c a l h a l f l i f e
f o r the r e l e a s e of testosterone and i t s esters from an
I.M. o i l i n j e c t i o n was c l o s e l y r e l a t e d to the o i l / w a t e r
d i s t r i b u t i o n c o e f f i c i e n t of the d e r i v a t i v e s . The o i l
used by James ejb a l . ( 2 6 4 ) was e t h y l o l e a t e . I t was
i n t e r e s t i n g to note that the homologous s e r i e s of f o r myl through v a l e r y l esters of testosterone had approximately equal s o l u b i l i t y i n the e t h y l oleate showing
that the d i s t r i b u t i o n c o e f f i c i e n t was l a r g e l y determined by the decrease i n water s o l u b i l i t y . Apart from
these i n t e r e s t i n g b a s i c s t u d i e s , the r e s u l t s of many
workers have shown that longer duration of a c t i o n of
testosterone from an I.M. i n j e c t i o n could be e f f e c t e d
by a c y l a t i o n of the 173-hydroxy group ( 2 6 5 - 2 8 8 ) .
Increasing chain length of the a c y l group e f f e c t i v e l y i n creased the d u r a t i o n of a c t i o n , i . e . , the testosterone
esters are thought to gradually leach out of the I.M.
i n j e c t i o n s i t e ( o i l based I.M. i n j e c t i o n ) , regenerate
testosterone i n the general c i r c u l a t i o n which then
exerts i t s androgenic a c t i v i t y ( 2 6 4 ) .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

53

Miescher ert a l . ( 2 6 5 ) were the f i r s t t o promote


the use of long chain esters o f testosterone as depot
forms of testosterone. Ott et^ a l . ( 2 7 6 ) have suggest
ed that the -cyclopentylpropionate ester of t e s t o s
terone (XCII) i n j e c t e d I.M. i n cottonseed o i l was a
superior ester form of testosterone when compared i n a
s e r i e s of saturated and nonsaturated e s t e r s . This
conclusion was based on the r e l a t i v e growth of seminal
v e s i c l e s i n castrated r a t s a f t e r I.M. dosing of the
various e s t e r s . S i m i l a r l y , Meier and Tschopp ( 2 8 4 )
using the growth of the capon c r e s t as an i n d i c a t o r ,
showed that the undecylenate e s t e r of testosterone
(XCIII) was f a r superior i n d u r a t i o n of a c t i o n t o the
propionate, i s o b u t y r a t e and -valerate e s t e r s
Voss
(277) and Haack et
OR

0
(XCI)

-H

(XCIII)

-COC H

(XCIV)

-C0CH C0C H

-C0C H

(XCVIII)

(XCIX)

(C)

(XCII)
10

21

(XCV)
(XCVI)

(XCVII)
C

6 i3
H

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

54

PRO-DRUGS

ketonic esters of t e s t o s t e r o n e , e s p e c i a l l y t e s t o s t e r one decanoylacetate (XCIV). Dekanski and Chapman ( 2 7 3 )


f e l t that testosterone phenyl propionate (XCV) was as
e f f e c t i v e as XCI but was f a r superior t o the e a r l i e r
promoted testosterone propionate (XCVI). D i c z f a l u s y
et a l . ( 2 7 1 ) demonstrated the s u p e r i o r i t y of a s e r i e s
of alkoxyhydrocinnamic a c i d esters of testosterone t o
both XCV and XCVI.
Para-heptyloxyhydrocinnamyltestosterone (XCVII), showing considerable a c t i v i t y over 90
days, appeared superior t o XCV and XCVI.
The study of Junkmann and W i t z e l ( 2 6 7 ) i s the
most comprehensive review of depot forms of t e s t o s t e r one and other s t e r o i d s . The enanthate (XCVIII), undecanoate ( X C I I I ) , and palmitat (XCIX)
f
terone a l l showed e x c e l l e n
E v a l u a t i o n of various other esters of testosterone
as depot forms of testosterone can be found ( 2 6 5 - 2 8 8 ) .
I t i s i n t e r e s t i n g t o note that Kishimoto ( 2 8 9 ) has r e c e n t l y noted the presence of enzymes i n the CNS that
are capable of s y n t h e s i z i n g f a t t y a c i d esters of t e s tosterone. McEwen et. a l . ( 2 9 0 ) have suggested that d i hydrotestosterone i s the a c t i v e form of testosterone.
I t does appear that testosterone esters are l e g i t i m a t e
pro-drugs of testosterone. However, because the major
e v a l u a t i o n of t h e i r release c h a r a c t e r i s t i c s i s based on
some pharmacological e f f e c t , some minor doubt does
e x i s t as t o whether the esters are t r u l y pro-drugs.
Junkmann and W i t z e l ( 2 6 7 ) document the various
testosterone esters a v a i l a b l e commercially i n Europe.
In the United S t a t e s , XCII, XCVI, XCVIII and t e s t o s terone phenylacetate (C) are a l l commercially a v a i l a b l e
as depot forms of testosterone.
Nandrolone (nortestosterone, CI) and various nandrolone d e r i v a t i v e s have a l s o been e s t e r i f i e d f o r the
purpose of prolonging the a c t i o n of these anabolic
agents a f t e r t h e i r S.C. or I.M. i n j e c t i o n i n an o i l
v e h i c l e ( 2 9 1 - 2 9 5 * 2 6 7 * 2 7 1 ) Nandrolone phenylpropionate
(CII) and nandrolone decanoate (CIII) are both commerc i a l l y a v a i l a b l e . C I I I i s longer a c t i n g than C I I and
i s administered monthly whereas C I I I i s administered
weekly ( 2 9 6 ) . The longer a c t i v i t y of C I I I (myotrophic
e f f e c t s as measured by seminal v e s i c l e growth i n r a t s )
with a s i n g l e 4 mg I.M. dose i n sesame o i l r e l a t i v e t o
CII ( s i m i l a r l y administered) was demonstrated by de
V i s s e r and Overbeek ( 2 9 5 ) . The a c t i o n of CII and the
mechanisms of anabolic androgenic a c t i v i t y were l a t t e r
discussed by van der Vies ( 2 9 5 ) A q u a n t i t a t i v e s t r u c ture anabolic a c t i v i t y a n a l y s i s of a s e r i e s of nandrolone esters has r e c e n t l y been presented by Chaudry and
James ( 2 9 4 ) while Pala et a l . ( 2 9 1 ) t e s t e d terpenoates

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

55

of nandrolone f o r anabolic a c t i v i t y .
Depot forms (I.M. i n j e c t i o n i n an o i l v e h i c l e ) of
other s t e r o i d s are a l s o quite common. The weekly use
of dromostanolone propionate i n the treatment of
breast carcinoma was discussed by Seay e_t a l . (297)
Long a c t i n g estrogenic s t e r o i d s u s e f u l i n the t r e a t ment of estrogen d e f i c i e n c y i n women has been a goal
of many workers. D e r i v a t i v e s of e s t r a d i o l (CIV), such
as the oligemeric d e r i v a t i v e s (CV longest a c t i n g der i v a t i v e ) of Kuhl ert a l . (298), p o l y e s t r a d i o l phosphate (CVI) of D i c z f a l u s y et a l . (299), e s t r a d i o l 3benzoate-17-cyclooctenyl ether (CVII) of P a l c o n i et
a l . (300), 3- and/or 17-acyl d e r i v a t i v e s of e s t r a d i o l
of F e r r i n (301), and others (302-310) were a l l shown
to have prolonged estrogeni
t r a t i o n . CVI was a l s
treatment of p r o s t a t i c carcinoma. Prolonged release
forms of dihydroxy progesterone f o r b i r t h c o n t r o l
(acetophenide d e r i v a t i v e s ) have been found u s e f u l
(311-313) while the caproate e s t e r of hydroxyprogesterone (Delalutin) i s used as a long a c t i n g s t e r o i d
i n amonorrhea (314).
The use of desoxycorticosterone acetate and t r i methylacetate i n adrenal i n s u f f i c i e n c y (Addison's d i s ease) has proven s u c c e s s f u l (315)
Desoxycorticosterone t r i m e t h y l a c e t a t e (CIX, Percorten p i v a l a t e ) has
a very prolonged a c t i o n and should not be administered
more than once a month (316).
Long a c t i n g , pro-drug r e p o s i t o r y i n j e c t a b l e forms
of c o r t i c o s t e r o i d s u s e f u l i n the treatment of i n f l a m mation are betamethasone acetate, methylprednisolone
acetate (Depo-Medrol), f l u o r o c o r t i s o n e acetate ( F l o r inef a c e t a t e ) , hydrocortisone cypionate (Cortef) and
triamcinolone hexacetonide (Aristocort), to name a
few

(316).

Winter et a l . (317) found that the r e l e a s e of the


parent s t e r o i d i n t o general c i r c u l a t i o n was a f u n c t i o n
not only of the p h y s i c a l p r o p e r t i e s of the pro-drug
( a f f e c t i n g the r e l e a s e from the i n j e c t i o n s i t e ) but
a l s o the chemical p r o p e r t i e s of the pro-drug ( a f f e c t ing the regeneration r a t e of s t e r o i d once released
from the i n j e c t i o n s i t e ) .
Fluphenazine (CX) d i h y d r o c h l o r i d e i s a drug usef u l i n the c o n t r o l of psychotic behavior and i s administered o r a l l y or by I.M. i n j e c t i o n (318.). Patient
compliance with a n t i p s y c h o t i c drugs i s a r e a l problem
(319). Fluphenazine enanthate (CXI) and fluphenazine
decanoate (CXII) are fluphenazine esters given by I.M.
i n j e c t i o n ( i n sesame o i l v e h i c l e ) which have prolonged
a c t i v i t y f o r up to two and four weeks (318,320-324).

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

(CVI): R = e s t r a d i o l
molecule

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

57

Depot forms of other n e u r o l e p t i c drugs e f f e c t e d by es


t e r i f i c a t i o n (325-331) and I.M. dosing i n an o i l v e h i
c l e are -fluphenthixol ( C X I I I ) , as i t s decanoate e s t e r
(CXIV), and p i p o t h i a z i n e (CXV) as i t s palmitate (CXVI)
and undecanoate esters (CXVII).
The prolongation of n e u r o l e p t i c a c t i v i t y of CXIV
i n Viscoleo administered I.M. compared to CXIII dihyd r o c h l o r i d e was demonstrated by Nymark et^ a l . (325)
Figure 7 compares the i n h i b i t i o n of a c o n d i t i o n a l

TIME, (DAYS)
Acta Pharmacologica et Toxicologica

Figure 7. Days after drug administrationsingle I.M.


dose of CXIV in oil (Q) or CXIII oral daily ( ) (325)

avoidance response a f t e r o r a l CXIII d i h y d r o c h l o r i d e


administered once d a i l y (5 mg/Kg) to CXIV (10 mg/Kg)
a f t e r a s i n g l e I.M. i n j e c t i o n i n o i l . Dreyfus et a l .
(323) i n the case of CX e s t e r s , V i l l e n e u v e et a l . 1*327)
i n the case of CXV e s t e r s , and Jorgensen et a l . (326)
i n the case of CXIII esters were able to show that the
metabolic p a t t e r n of the esters was i d e n t i c a l to those

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

PRO-DRUGS

58

(CX):

R = -H

(CXI)
(CXII): R = - C O C H
n

(CXIII): R = -H
(CXIV):

R = -COC H
Q

C H CH C H /
2

\ c H CH 0 R

S0 N(CH )
2

(CXV):

R = -H

(CXVI): R = - C O C
(CXVII):

1 5

R = -C0C

i n

3 1

o l

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

59

Overview and Definition

STELLA

(CXIX)
(CXX):

R = -COC H
15

31

of the parent compounds and that the a c t i v i t y of the


e s t e r s appeared to be r e l a t e d to the formation of the
parent n e u r o l e p t i c .
Another p a r e n t e r a l r e p o s i t o r y pro-drug e f f e c t e d
through e s t e r formation was O-palmitoylamodiaquine
(CXX), a moderately s u c c e s s f u l depot form of amodiaquine (CXIX) (332). E l s l a g e r (333), i n h i s review of
chemotherapy i n the treatment of m a l a r i a and i n p a r t i c
u l a r r e p o s i t o r y forms of a n t i m a l a r i a l drugs, discussed
at great lengths v a r i o u s means by which the d u r a t i o n of
a c t i o n of v a r i o u s a n t i m a l a r i a l agents could be extend
ed. The two most i n t e r e s t i n g examples given were v a r i
ous a c y l a t e d and S c h i f f base forms of 4,4'-diaminodiphenylsulfone (LXXXVIII) and the use of s p a r i n g l y water
s o l u b l e s a l t s of c y c l o g u a n i l (CXXI) and chloroguanide
(CXXII).
4,4 -Diacetoamidodiphenylsulfone (CXXIII) was
found to give a r e p o s i t o r y a n t i m a l a r i a l e f f e c t when i n
j e c t e d I.M. to P. berghei i n f e c t e d mice. However,
CXXIII had poor a c t i v i t y i n r a t s . The poor a c t i v i t y
i n r a t s was a t t r i b u t e d by Thompson (334) to the i n a b i l
i t y of r a t s to metabolize CXXIII to LXXXVIII whereas
f

0
CH CHN-^/
3

(CXXIII)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

60

PRO-DRUGS

mice were able t o convert CXXIII t o LXXXVIII. CXXIII


was found to be u s e f u l i n the treatment of P. f a l i c i arum i n humans f o r 42 days a f t e r a s i n g l e 3-25 mg/Kg
I.M. i n j e c t i o n (333).
As noted i n the s t e r o i d examples, the a b i l i t y of
a drug t o leave an I.M. i n j e c t i o n s i t e i s h i g h l y depen
dent on the s o l u b i l i t y of the drug i n p h y s i o l o g i c a l
f l u i d s and i n the i n j e c t i o n v e h i c l e . I f the drug i s
water i n s o l u b l e , an I.M. i n j e c t i o n of a suspension of
the drug w i l l deposit i n the muscle and over a period
of time gradually d i s s o l v e s , r e l e a s i n g the drug i n t o
general c i r c u l a t i o n . The aqueous s o l u b i l i t y of any
amine s a l t i s dependent on the counter anion with the
s o l u b i l i t y equal t o the square root of the s o l u b i l i t y
product of the s a l t
the s a l t i s h i g h l y p dependent

Cl

(CXXII)

(CXXI)

The poor i n v i t r o a c t i v i t y of chloroguanide


(CXXII) compared to i t s good i n v i v o a c t i v i t y suggested
that CXXII was metabolized i n the body t o an a c t i v e
metabolite (335-340). This a c t i v e metabolite was found
to be c y c l o g u a n i l (CXXI) which has a short d u r a t i o n of
a c t i o n because of r a p i d e x c r e t i o n (333)> Various spari n g l y water soluble s a l t s of CXXII were found t o be
poor r e p o s i t o r y forms of CXXII but the pamoate s a l t of
CXXI (CXXIV) with an aqueous s o l u b i l i t y of 0.03 mg/ml
at pH 7 was found t o be e f f e c t i v e against P. berghei i n
i n f e c t e d mice f o r up t o 8 1/2 weeks when given S.C.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

Overview and Definition

STELLA

61

(333). Figure 8 shows a p l o t of the logarithm PMW,


the estimated weeks 50% of the mice were protected from
challenge w i t h P. b e r g h e i against logarithm S, the
s o l u b i l i t y of various s a l t s of CXXI. The s o l i d l i n e i s
s

oc
a

tu
H

LU
I-

- S
Ui

ce
Lu

g *
X

Lu
LU

LU

8"

|i
CO
Lu

Figure 8. Plot of log PMW vs. log (solubility) for a series of


CXXI salts. Plotted from the data of ElsUger (333).
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

62

PRO-DRUGS

(CXXIV)
the l e a s t squares l i n e described by equation 1.
l o g PMW

= -0.71

+ 0.11

l o g S + 0.077

The
(eq.

1)

c o r r e l a t i o n c o e f f i c i e n t was 0 . 9 0 6 . As i s r e a d i l y apparent, the l e s s water s o l u b l e s a l t s gave the greater


r e p o s i t o r y e f f e c t s confirming that the r a t e determining
step i n the r e l e a s e of CXXI from the S.C. i n j e c t i o n of
400 mg/Kg equivalent of CXXI was the d i s s o l u t i o n of the
sparingly soluble s a l t s .
For a comprehensive review of r e p o s i t o r y s a l t , est e r and amide forms of various other a n t i m a l a r i a l s , the
reader i s d i r e c t e d to the review by E l s l a g e r (333)> a
s e r i e s of papers by E l s l a g e r et a l . ( 3 3 2 , 3 4 1 - 3 4 4 ) , and
the references t h e r e i n .
Recently naloxone (CXXV), used as a n a r c o t i c ant a g o n i s t , was found to be very potent but e f f e c t i v e f o r
only three to four hours i f administered p a r e n t e r a l l y
(238-241). Levine et a l . (346) found that by g i v i n g
I.M. i n j e c t i o n s of a.suspension of naloxone pamoate
(CXXVI) a sustained r e l e a s e a c t i o n was seen which
blocked the e f f e c t of opiates without harmful side e f f e c t s f o r up to 72 hours. Use of s p a r i n g l y s o l u b l e
a c i d s a l t s to e f f e c t sustained r e l e a s e a f t e r I.M. pare n t e r a l a d m i n i s t r a t i o n w i t h a number of other amines
such as streptomycin (347*348) dihydrostreptomycin
(347)s naltrexone (349TT~eyelazocine ( 3 5 0 ) , and others
(351) have proven to be r a t h e r s u c c e s s f u l . Thompson
and Hecht (352) prepared and demonstrated the sustained

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

63
r

r e l e a s e of cyanocobalamine (CXXVII) or v i t a m i n B^2 ? om


cyanocobolamine zinc tannate (CXXVIII). Figure 9 shows
the serum l e v e l s of CXXVII a f t e r I.M. i n j e c t i o n of 500
\ig of CXXVII i n normal s a l i n e compared to serum l e v e l s
of CXXVII from a molar equivalent amount of CXXVIII.

ol

Li

12

15

18

21

24

27

30

DAY

AFTER INJECTION
American Journal of Clinical Nutrition

Figure 9. CXXVII serum levels in humans after I.M in


jection of 500 mg of CXXVII ( ) and a molar equivalent
amount of CXXVIII (Q) (352)

P a r e n t e r a l l y sustained r e l e a s e of a c i d substances
such as p e n i c i l l i n has s i m i l a r l y been e f f e c t e d by the
use of s p a r i n g l y soluble s a l t s of p e n i c i l l i n such as
benzathine and procaine p e n i c i l l i n (7-11). The spar-

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

64

PRO-DRUGS

i n g l y soluble protamine z i n c i n s u l i n might a l s o be considered as a chemical m o d i f i c a t i o n u s e f u l i n e f f e c t i n g


sustained r e l e a s e (353) of i n s u l i n .
Attempts to produce sustained r e l e a s e or prolonged
a c t i o n medication f o r o r a l a d m i n i s t r a t i o n by the use of
s p a r i n g l y water s o l u b l e s a l t s have been m a r g i n a l l y succ e s s f u l . Quinidine polygalacturonate does produce a
prolonged r e l e a s e q u i n i d i n e on o r a l a d m i n i s t r a t i o n
(345).
The marginal success of these types of products
stems from the f a c t that the minimum s o l u b i l i t y (or
maximum s t a b i l i t y ) of the s a l t s occurs around n e u t r a l i t y . Because the GI t r a c t has a wide spectrum of pH
(from the strong a c i d of the stomach to the s l i g h t l y
a l k a l i n e lower i n t e s t i n e )
f th comple s a l t
( i n c l u d i n g r e s i n s ) ten
r a p i d l y than the e q u i l i b r i u
y
(or r e s i n s ) might i n d i c a t e . Marginal prolonged r e l e a s e
e f f e c t s were noted with codeine r e s i n a t e ( 3 5 4 ) and amphetamine r e s i n a t e (355) M i l l e r et a l . ( 3 5 6 ) i n a
c l i n i c a l study appeared to confirm that a" s i n g l e 75 nig
dose of imipramine pamoate was t h e r a p e u t i c a l l y equival e n t to d i v i d e d doses of 25 mg three times a day of
imipramine hydrochloride. Other examples are amphetamine tannate ( 3 5 7 ) * p y r a n t e l pamoate ( 3 5 8 , 3 5 9 ) * pamoates
( 3 6 0 ) , tanntes (351)
and rsintes ( 3 6 1 , 3 6 2 ) of various
amine drugs.
An i n t e r e s t i n g study by Loucas and Haddad (363)
demonstrated that p i l o c a r p i n e a l g i n a t e , a s p a r i n g l y
water s o l u b l e s a l t of p i l o c a r p i n e , when a p p l i e d as s o l i d f l a k e s to the cul-de-sac of the eye could e f f e c t the
prolonged release of p i l o c a r p i n e . Another i n t e r e s t i n g
prolonged r e l e a s e pro-drug i s t r i a c e t i n (CXXIX), a dermal d e l i v e r y form of the f u n g i s t a t i c agent a c e t i c a c i d
( 3 6 4 ) . The d i r e c t percutaneous a p p l i c a t i o n of a c e t i c
acid i s too c o r r o s i v e and short a c t i n g while CXXIX
g r a d u a l l y releases a c e t i c a c i d i n noncorrosive q u a n t i t i e s . Dimethylol urea (CXXX) and 3 - m e t h y l o l - 5 , 5 - d i methylhydantoin (CXXXI) are two prolonged r e l e a s e forms
of the s k i n d i s i n f e c t a n t formaldehyde (CXXXII). Both
drugs, when a p p l i e d dermally, g r a d u a l l y release CXXXII
(364,365).

The use of polyethylene g l y c o l d e r i v a t i v e s


(CXXXIII) of procaine (CXXXIV) by Weiner and Z i l k h a
( 3 6 6 ) to prolong the l o c a l anesthetic a c t i o n of dermall y a p p l i e d procaine was s u c c e s s f u l . CXXXIII and the
equivalent d e r i v a t i v e with PEG 400 showed a slower onset of a c t i o n but an increase i n d u r a t i o n of a c t i o n .
The r e s u l t s were c o n s i s t e n t with CXXXIV being the act i v e agent r a t h e r than the i n t a c t CXXXIII.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

65

Overview and Definition

CH 0C0CH
I
CH-OCOCEL
CH OCOCH
o

-> CH COOH (FUNGISTATIC AGENT)


3

(CXXIX)

HOCH NHCNHCH OH
2

(CXXX)

H(?H

i n vivo

H
3

CH
c
I

(CXXXII)

C= 0
1
NCH OH
2

(CXXXI)

RCO(CH CH 0) CR
2

i|

R = NH

(CXXXIII)

(H C ) NCH CH OC
5

('

COCH CH N(C H )
2

NH

(CXXXIV)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

66

PRO-DRUGS

E s t e r i f i c a t i o n of i n s e c t r e p e l l e n t dihydroxyacetone increased the d u r a t i o n a c t i o n of t h i s dermally app l i e d product ( 3 6 7 ) * Garner et a l . ( 3 6 7 ) suggested


that the i n t a c t e s t e r s may have some a c t i v i t y of t h e i r
own and may not be a c t i n g as pro-drugs.
The Use of Pro-Drugs to Increase the Aqueous S o l u b i l i t y
of a Drug to Allow f o r E i t h e r D i r e c t Aqueous I.V. or
I.M. I n j e c t i o n , or Opthalmic D e l i v e r y
As we have already seen, many drugs possess very
poor aqueous s o l u b i l i t y which o f t e n leads to l i m i t e d
o r a l b i o a v a i l a b i l i t y . S i m i l a r l y , many drugs on o r a l
dosing undergo " f i r s t
" metabolism
P a r e n t e r a l ad
m i n i s t r a t i o n of drug
travenously or i n t r a m u s c u l a r l y
y advantages
(a) allows r a p i d blood l e v e l s of the drug to be obt a i n e d . This i s e s p e c i a l l y u s e f u l i n emergency t r e a t ments; (b) e f f i c i e n t d e l i v e r y of the drug, e s p e c i a l l y
f o r drug t e s t i n g ; and (c) allows d e l i v e r y of a drug
when o r a l therapy i s not f e a s i b l e .
A number of s p a r i n g l y water s o l u b l e drugs are adm i n i s t e r e d i n mixed organic/aqueous v e h i c l e s . I.M. i n j e c t i o n s of chlordiazepoxide ( 3 6 8 ) , diazepam ( 3 6 9 - 3 7 0 ) *
and sodium diphenylhydantoin ( 3 7 1 - 3 7 5 ) administered i n
propylene glycol/water v e h i c l e s have shown delayed abs o r p t i o n due to p r e c i p i t a t i o n of the administered drug
at the i n j e c t i o n s i t e . The delayed absorption w i t h
sodium diphenylhydantoin (I.M. dosing) l e d to increased
seizure r a t e s i n t r e a t e d p a t i e n t s ( 3 7 ^ )
As discussed e a r l i e r , s t e r o i d s are a group of compounds w i t h poor aqueous s o l u b i l i t y . Adrenal c o r t i c o s t e r o i d s such as betamethasone, dexamethasone, hydroc o r t i s o n e , methylprednisolone and prednisolone are a l l
a v a i l a b l e commercially as water s o l u b l e sodium phosphate esters or sodium hemisuccinate e s t e r s . These
soluble c o r t i c o s t e r o i d s are used i n emergency treatment
of b r o n c h i a l asthma ( s t a t u s asthmaticus), acute adrenal
c o r t i c a l i n s u f f i c i e n c y , a l l e r g i c drug r e a c t i o n s and are
given i n t r a a r t i c u l a r l y or i n t r a s y n o v i a l l y i n the t r e a t ment of j o i n t inflammation. The water s o l u b l e prodrugs (CXXXV as sodium phosphate, or CXXXVI as sodium
succinate) regenerate the parent s t e r o i d i n v i v o ( 3 7 9 )
CXXXV regenerates the parent s t e r o i d v i a a c i d and a l k a l i n e phosphatase enzymes while CXXXVI regenerates the
parent s t e r o i d v i a esterase enzymes.
Lange and S t e i n ( 3 7 6 ) synthesized other water s o l uble d e r i v a t i v e s of s t e r o i d s such as amino a c i d carbamates (CXXXVII). However, these d e r i v a t i v e s were found
to be e i t h e r i n a c t i v e or poorly a c t i v e .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

67

Overview and Definition

R-P-OH
(

Na

(CXXXV)

i n vivo

Na
( CXXXVI

where R = c o r t i c o s t e r o i d molecule
Although prednisolone hemisuccinate sodium s a l t
(CXXXIX) i s the current commercially a v a i l a b l e water
s o l u b l e form of prednisolone (CXXXVIII), other water
s o l u b l e d e r i v a t i v e s such as prednisolone-21-m-sulfobenzoate sodium s a l t (XCLI) and prednisolone-21-disodium
phosphate (XCL) have a l s o been synthesized and t e s t e d
(377-379). S i m i l a r l y , methylprednisolone sodium s u c c i
nate (316,380), hydrocortisone-21-phosphate (379,381),
-21-succinate (316,379*381-382), -21-aminoalkylcarboxyl a t e s (383), -21-m-sulfobenzoates (384) , and -21-sulf a t e s (384), dexamethasone sodium phosphate (316,379)*
betamethasone disodium phosphate (316) have been syn
t h e s i z e d and found to be water s o l u b l e , p a r e n t e r a l l y
b i o a v a i l a b l e forms of the parent s t e r o i d s .
The succinate e s t e r s of the various s t e r o i d s are
u s e f u l but s u f f e r somewhat from s t a b i l i t y problems,
i . e . , the drug must be s u p p l i e d as a l y o p h i l i z e d powder
f o r r e c o n s t i t u t i o n (316). Flynn et. a l . (385)> i n d i s
cussing the s o l v o l y s i s and f a c t o r s a f f e c t i n g the s t a
b i l i t y of corticosteroid-21-phosphate e s t e r s , s t a t e s
that " a d d i t i o n a l l y , some types of phosphate e s t e r s are
s u f f i c i e n t l y s t a b l e to allow the f o r m u l a t i o n of s o l u
t i o n s with p r a c t i c a l s h e l f - l i v e s . " Another added ad
vantage of phosphate e s t e r s as opposed to the succinate
esters i s t h e i r very r a p i d conversion to the parent
s t e r o i d . Melby et a l . (379) i n a very i n t e r e s t i n g
study showed that I.M. administered hydrocortisone-21phosphate ( X C L I I ) , (XCL), dexamethasone-21-phosphate
(XCLIII) and I.V. administered XCLII a l l produced
higher plasma l e v e l s of the parent s t e r o i d than the
corresponding 21-succinate e s t e r s , (see Figure 10).

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

68

PRO-DRUGS

0
RCNHCH CCP

(CXXXVII):

R = various s t e r o i d molecules

CH 0R
C=0
-OH
o

(CXXXVIII): R
(CXXXIX): R
(XCL): R

-COCH CH COO
2

-PO

Na

2Na

(XCLI): R =
so

Na

I t was suggested by Melby ejb a l . (379) that the 21-suc


cinate esters underwent metabolism at other points i n
the molecule before d e - e s t e r i f i c a t i o n . This could oc
cur i f d e - e s t e r i f i c a t i o n was slow r e l a t i v e to other
metabolic processes. The 21-phosphates, on the other
hand, undergo r a p i d dephosphorylation r e l e a s i n g the
parent s t e r o i d . Although Melby elb a l . s (379) data
does not suggest i t , another p o s s i b l e mechanism i s that
the 21-succinate might be excreted unchanged. The ac
t i v e e l i m i n a t i o n of a c i d i c drugs such as p e n i c i l l i n and
p r o b e n i c i d have been shown to occur (386) .
Sandman et a l . (387388) i n studying the aqueous
s t a b i l i t y of chloramphenicol succinate (XCLIV), a water
soluble pro-drug of chloramphenicol (XCLV) s u i t a b l e f o r
I.V., I.M. and opthalmic d e l i v e r y of chloramphenicol,
noted an unusual p a r t i a l a c y l t r a n s f e r r e a c t i o n (see
scheme IX) of the s u c c i n y l group to give a c y c l i c hemiortho ester (XCLVI). A s i m i l a r r e a c t i o n may a l s o po
t e n t i a l l y occur f o r hydrocortisone (see Scheme X).
f

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

69

XCLIX, i f formed, would be more s t a b l e , i . e . , would not


be subjected to esterase a c t i v i t y , and might be e l i m i nated or metabolized v i a other pathways.

250

TIME (minutes)
Metabolism

Figure 10. Mean 17,21 -dihydroxy-20-oxosteroids in


plasma following I.M. injection of XCLII ( ), LV. injection of XCLII (), I.M. injection of hydrocortisone-21 succinate (0)> &nd LV. injection of hydrocortisone-21succinate (), in a dose of lmg/kg of body weight in
humans (379)

The estrogenic s t e r o i d , d i e t h y l s t i l b e s t e r o l (CL),


may be given I.V. as i t s diphosphate e s t e r (CLI) d i s o dium s a l t i n the treatment of p r o s t a t i c carcinoma.
However, the o r i g i n a l synthesis of CLI was not done
with the i n t e n t i o n of obtaining a water s o l u b l e form of
CL but to help l o c a l i z e CL i n the carcinoma c e l l s by
u t i l i z i n g the high a c i d and a l k a l i n e phosphatase l e v e l s
(389-397) i n carcinoma c e l l s to e f f e c t p r e f e r e n t i a l uptake of CL. Water soluble g l y c i n e esters of CL have

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

70

PRO-DRUGS

0
Il

0 A

I I
2

i
C=0
I
CHC1

H
2 2

(XCLIV)

i n vivo

CH CH COO*
Na
2

CHCHCH 0H
I I 2
OHNH
I
c=o
I
CHC1
o

^CH
I

NH
I

(XCLV)

CHC1
(XCLVI)

Scheme IX

(XCLVIII)

(XCLIX)

Scheme X

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

71

Overview and Definition

a l s o been synthesized (398.).


Hydroxydione (CLII) i s a water i n s o l u b l e b a s a l
anesthetic that may be given I.V. (398) as i t s sodium
succinate d e r i v a t i v e ( C L I I I ) .
0

(CLII) : R = -H
( C L I I I ) : R = -COCH CH COO Na
0

As stated e a r l i e r , chloramphenicol (XCLV), a


s l i g h t l y water s o l u b l e a n t i b i o t i c , cannot be used d i
r e c t l y f o r I.V., or I.M. i n j e c t i o n or f o r l o c a l s o l u
t i o n a p p l i c a t i o n as eye/ear drops. Glazko ejt a l .
(399) overcame t h i s problem by s y n t h e s i z i n g the sodium
s a l t of chloramphenicol monosuccinate (XCLIV). XCLIV
can be given I.V./I.M. as a r e c o n s t i t u t e d i n j e c t a b l e
and i s reasonably q u a n t i t a v e l y converted to XCLV and
s u c c i n i c a c i d (399) by esterase a c t i v i t y present i n
plasma (400-402). Adenine arabinoside (CLIV), an an
t i v i r a l , c y t o t o x i c agent whose low aqueous s o l u b i l i t y
prevented i t s use i n small volume I.V. p r e p a r a t i o n s ,
was s o l u b i l i z e d by p r e p a r a t i o n of the 5'-formate e s t e r
(CLV) (see paper by Dr. Repta). LePage et a l . (403.)
prepared the 5 -phosphate (CLVI) of CLIV. However,
CLVI may undergo deamination before regenerating CLIV
which may l i m i t i t s usefulness (4344).
Oxazepam (CLVII), a minor t r a n q u i l i z e r and a c t u a l
l y a metabolite of diazepam (405-408), has been s o l u
b i l i z e d as oxazepam sodium succinate ( C L V I I I ) . CLVIII
i s a pro-drug of CLVII with favorable p h y s i c a l proper
t i e s f o r I.V. or I.M. a d m i n i s t r a t i o n (409-413).
An
i n t e r e s t i n g side l i n e to t h i s work i s that the number
3 carbon of CLVII i s an o p t i c a l l y a c t i v e center and
the two p o s s i b l e isomers of CLVIII have been shown t o
regenerate CLVII at d i f f e r i n g r a t e s (410). The d i f f

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

72

PRO-DRUGS

(CLIV)
(CLV): R = -COH
9

(CLVI) : R = -P0 H Na'


o

(CLVII): R = -H
8

(CLVIII): R = -COCHgCHgCOoSla

f e r i n g r a t e s were also found t o be a f u n c t i o n of the


p a r t i c u l a r animal species i n which the regeneration was
studied (410).
Recently S t e l l a and Higuchi (4l4) synthesized and
demonstrated the p o s s i b l e usefulness of 3 N ,N -diethylaminoethyl-5-methyl-2,2-ethylphenylhydantoate
(CLIX)
as a water s o l u b l e pro-drug of mephenytoin (CLX).
CLIX i s converted to CLX under p h y s i o l o g i c a l c o n d i t i o n s
without enzyme mediation. C u r r e n t l y the only i n j e c t able anticonvulsant hydantoin a v a i l a b l e i s sodium d i phenylhydantoin which, due to i t s a l k a l i n i t y , has many
unwanted side e f f e c t s (415). A d e r i v a t i v e of 5,5-diphenylhydantoin s i m i l a r to CLIX w i l l be reported by
S t e l l a et a l . l a t e r .
t

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

73

Overview and Definition

B i o r e v e r s i b l e chemical m o d i f i c a t i o n of drug mole


cules t o increase the aqueous s o l u b i l i t y o f a compound
has a l s o been u t i l i z e d i n s o l u b i l i z i n g menadione,
e i t h e r as i t s b i s u l f i t e adduct, i t s disodium diphos
phate e s t e r , or i t s carboxymethoxime d e r i v a t i v e ( 4 l 6 4 l 8 ) , tetrahydrocannabinol as a s e r i e s of b i f u n c t i o n a l
esters (419-421), and -methydopa as i t s e t h y l ester
(422) .

HN\

(CLIX)
in vivo/in vitro

-c=o

HC5

CH^
II

0
(CLX)

The Use of Pro-Drugs t o Lower the T o x i c i t y of a Drug


A drug may e x h i b i t t o x i c i t y i f i t accumulates
s e l e c t i v e l y i n some t i s s u e or organ and i n t e r a c t s with
some receptor i n the organ thus e l i c i t i n g a r e a c t i o n
which i s not n e c e s s a r i l y the desired pharmacological
r e a c t i o n . Any drug-receptor i n t e r a c t i o n w i l l be depen
dent on the amount of drug reaching the receptor and
i t s time p r o f i l e i n contact with the receptor.
As
stated e a r l i e r , "peak" and " v a l l e y " e f f e c t s of m u l t i p l e
dosing might r a i s e the concentration of a drug i n the
plasma above i t s t o x i c l e v e l o r , at the end of i t s

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

74

PRO-DRUGS

c y c l e (before the next dose i s g i v e n ) , the plasma l e v e l


may drop below the therapeutic l e v e l . For these r e a sons, a sustained r e l e a s e or prolonged release form of
a drug may be d e s i r a b l e to narrow the d i f f e r e n c e seen
between the "peaks" and " v a l l e y s . "
I f a pro-drug w i t h
i n t r i n s i c a l l y lower t o x i c i t y i s designed to regenerate
the parent compound slowly (the equivalent to slow absorption) the t o x i c i t y of the parent compound might be
lowered.
A few i n t e r e s t i n g , although c o n t r o v e r s i a l , and
perhaps marginal examples of t h i s phenomena do e x i s t .
The agent 5,5-ethylphenylhydantoin (CLXI), was used as
an a n t i c o n v u l s a n t . I t was removed from the market because of t o x i c i t y and i n i t s place mephenytoin (CLX)
was promoted (423)
i s N-demethylated t
althoug
out t o x i c i t y i t i s s t i l l used i n the treatment of p e t i t
mal s e i z u r e s (423) The known a c t i v i t y of CLX, which
does not i n v o l v e any l a g time, i s c o n s i s t e n t with CLX
maintaining some anticonvulsant a c t i v i t y of i t s own.
S i m i l a r l y primacolone (primidone, CLXII) was promoted as a nontoxic anticonvulsant u s e f u l i n the t r e a t ment of grand mal s e i z u r e s (424). S p e c i f i c a l l y , i t was
f e l t that i t might replace phenobarbitone ( C L X I I I ) . I t
has since been found that CLXII i s m e t a b o l i c a l l y o x i d i z e d to CLXIII both i n humans and animals (426-428).
Kutt (425) i n a recent review of pharmacodynamic and

NH
.NH
(CLXII)

(CLXIII)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

OH

1.

STELLA

Overview and Definition

75

pharmacokinetic measurements of a n t i e p i l e p t i c drugs


states that CLXIII plasma l e v e l s a f t e r chronic admini s t r a t i o n of CLXII are 1.5-3 times higher than the
CLXII l e v e l s . This i s c o n s i s t e n t with the slow e l i m i n a t i o n of CLXIII. Even though the long term a n t i e p i l e p t i c e f f e c t s of CLXII might be a t t r i b u t e d to i t s
m e t a b o l i t e , CLXIII, i t does appear that CLXII and another metabolite phenylethylmalonamide have a n i t e p i l e p t i c a c t i v i t y of t h e i r own (425)*
Another drug whose t o x i c i t y appears r e l a t e d to i t s
t i s s u e d i s t r i b u t i o n i s the c y t o t o x i c agent adriamycin
(CLXIV). The c a r d i o t o x i c i t y of CLXIV has been w e l l est a b l i s h e d . Arcamone et a l . (429) synthesized and
showed the change i
d i s t r i b u t i o i mic hear
t i s s u e a f t e r the a d m i n i s t r a t i o
tanoate (CLXV). CLXV was shown to have comparable
c y t o x i c a c t i v i t y to CLXIV. A f t e r CLXV dosing of t r i tium l a b e l l e d compound, CLXV showed greater amounts of
r a d i o l a b e l l e d m a t e r i a l i n lungs, l i v e r , and spleen
r e l a t i v e to CLXIV dosed animals, but lower amounts of
r a d i o l a b e l l e d m a t e r i a l s were seen i n the heart and
kidney.
Probably the area of drug t o x i c i t y which has been
given the greatest a t t e n t i o n and f o r which the pro-drug
approach has o f t e n been used i s g a s t r i c i r r i t a t i o n .
Drug induced g a s t r i c u l c e r a t i o n has been a recognized

OCH.

'3

OH
0

NH
(CLXIV): R
(CLXV): R

-H
-COC^H

7"15

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

76

PRO-DRUGS

problem e s p e c i a l l y i n p a t i e n t s t r e a t e d f o r inflammatory
diseases. S a l i c y l a t e s i n c l u d i n g a c e t y l s a l i c y l i c a c i d
or a s p i r i n , as w e l l as s a l i c y l i c a c i d i t s e l f , have been
known to induce g a s t r i c bleeding, u l c e r a t i o n and genera l GI i r r i t a t i o n (430-441). Attempts to modify s a l i c y l i c and a c e t y l s a l i c y l i c a c i d i r r i t a t i o n by the prodrug approach has a long h i s t o r y . None of the products
studied appears to hold any great advantage over acet y l s a l i c y l i c a c i d e i t h e r i n a c t i v i t y or decreased gast r i c i r r i t a n t properties.
However, p o t e n t i a l decreased
g a s t r i c i r r i t a n t properties with the use of some of the
pro-drugs have been claimed by some of t h e i r promoters
(442-445).

Other s a l i c y l i
benzoi a c i d e r i v a t i v e whic
have shown g a s t r i c i r r i t a n
c a l l y modified i n attempts to decrease t h e i r i r r i t a n t
properties.
The g l y c e r y l ester of N - a r y l a n t h r a n i l i c
a c i d apparently has lower g a s t r i c i r r i t a n t p r o p e r t i e s
than the parent compound (446), while various b i o r e v e r s i b l e d e r i v a t i v e s of n i c o t i n i c a c i d showed no advantage or showed marginal advantages over n i c o t i n i c a c i d
(447-448).

A l l potent n o n s t e r o i d a l anti-inflammatory agents


appear to e x h i b i t GI i r r i t a n t p r o p e r t i e s .
Indomethicin
(CLXVI) has been p a r t i c u l a r l y notorious.
The a c i d i c
p r o p e r t i e s of the anti-inflammatory agents and GI i r r i tants i n general suggest that the b i o r e v e r s i b l e b l o c k ing of the a c i d f u n c t i o n may lead to a decrease i n GI
i r r i t a t i o n . This i s not to say that t h e i r i r r i t a n t
mechanism of a c t i o n i s l o c a l because i t has been shown
that p a r e n t e r a l a d m i n i s t r a t i o n of many of these agents
does promote u l c e r a t i o n * However, i t also stands to
reason that high l o c a l i z a t i o n i n the GI mucosa of these^
agents on o r a l a d m i n i s t r a t i o n can only help promote
rapid ulceration.
Glamkowski et_ al_. (449), although not s t a t i n g that
t h e i r i n t e n t i o n was to bypass the g a s t r i c i r r i t a n t
p r o p e r t i e s of CLXVI, tested the a c t i v i t y of the aldehyde analog of CLXVI. Using the canageenan-induced
foot odema t e s t i n the r a t , the aldehyde analog (CXVII)
was 0.6-0.7 times as a c t i v e as CLXVI. Subsequent a n a l y s i s of plasma showed that CLXVII gave s u b s t a n t i a l
plasma l e v e l s of CLXVI presumably as a r e s u l t of oxidat i v e metabolism (see F i g u r e 11).
Whether the lower
CLXVI l e v e l s a f t e r CLXVII dosing i s due to incomplete
absorption or incomplete metabolism i s u n c e r t a i n .
The
authors d i d not state whether any d i f f e r e n c e s i n GI i r r i t a n t p r o p e r t i e s between the two compounds e x i s t e d .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

77

Figure 11. CLXVI plasma level-time profile after


administration of CLXVI (O) d CLXVII () at a
dose of 10 mg/kg orally to rats. Plotted from the
data of Glamkowski et al. (449)
an

(CLXVI): R = -COOH
(CLXVII): R =

-COH

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

78

PRO-DRUGS

Phenylbutazone (CLXVIII), another a c i d (carbon


acid) n o n s t e r o i d a l agent, has been shown to be a GI i r r i t a n t . I t has yet to be e s t a b l i s h e d whether the 0trimethoxybenzoyl enol e s t e r of CLXVIII (CLXIX), which
supposedly i s b e t t e r t o l e r a t e d and l e s s t o x i c than
CLXVIII (450), or an a c y c l i c form of CLXVIII (CLXX),
which i s p a r t i a l l y metabolized to CLXVIII (451-454),
hold any great advantage over CLXVIII.
C l o f i b r a t e or p-chlorophenoxyisobutyric a c i d e t h y l
e s t e r (CLXXI) synthesized by Jones ejb a l . (455) was
found to be u s e f u l as an antihypercholesteremic agent.
I t i s g e n e r a l l y accepted that the corresponding a c i d ,
p-chlorophenoxyisobutyric a c i d (CLXXII), i s the pharmac o l o g i c a l l y a c t i v e specie (456)
Th
f th e t h y l
e s t e r as opposed to
pears r e l a t e d to the b e t t e r tolerance of CLXXI on pro
longed a d m i n i s t r a t i o n (457).
Oleandrin, a potent d i u r e t i c u s e f u l i n the t r e a t ment of cardiac i n s u f f i c i e n c y , was e s t e r i f i e d to i t s

0
II

(CLXIX)

Ca ++

2
(CLXX)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

79

acetate ester i n an attempt to lower i t s GI d i s t u r bance tendencies (458-459).


Pain due to I.M. i n j e c t i o n i s a t o x i c i t y problem
that has been associated with a number of compounds.
Intramuscular i n j e c t i o n s of the a n t i b i o t i c , clindamycin, were found to be p a i n f u l but a phosphate ester prodrug of clindamycin on I.M. dosing (460-461) was p a i n l e s s . S i m i l a r l y chloramphenicol sodium succinate (399)
and b i o r e v e r s i b l e oleandomycin d e r i v a t i v e s (168,172173) were l e s s i r r i t a t i n g on I.M. i n j e c t i o n than the
parent compounds.
Subcutaneous and I.M. i n j e c t i o n of i o n i c i r o n and
c o l l o i d a l i r o n were found to be e i t h e r t o x i c or very
p a i n f u l . Martin et a l (462) and others (463-466)
found that i r o n administere
(Imferon) was w e l l
low t o x i c i t y and i r r i t a t i o n , and was a s a t i s f a c t o r y
hematinic agent. Complexes of i r o n with s o r b i t a l (467)
have also proven s u c c e s s f u l . Terrato et a l . (468-470)
have studied o r a l i r o n absorption from complexes with
low molecular weight noncarbohydrate polymers and
shown that the absorption i s enhanced i n the presence
of these c h e l a t i n g agents. In the body, i r o n i s
stored as the complex f e r r i t i n and i s transported i n
combination with the p r o t e i n B - g l o b u l i n . The use of
polysaccharide molecules such as dextran, e t c . , was an
attempt to simulate the n a t u r a l l y occurring macromolecules as a transport medium f o r i r o n .
Two general examples where the pro-drug approach
has l e d to a reduction i n t o x i c i t y but where the mechanisms are not understood are amphotericin methyl est e r (CLXXIII) and the e t h y l ester of p r o s t a g l a n d i n .
Bonner et a l . (471) tested both the a c t i v i t y and t o x i c i t y of a s e r i e s of polyene a n t i b i o t i c esters and found
that the t o x i c i t y of the esters was much lower than
the t o x i c i t y of the parent compounds. Amphotericin
(CLXXIV) i s administered as a large volume I.V. c o l l o i d
suspension. The z w i t t e r i o n i c nature of CLXXIV i s a t t e s t e d to by i t s low water s o l u b i l i t y at n e u t r a l pH
and i n c r e a s i n g s o l u b i l i t y below pH 2 and above pH 11
(472) . CLXXIII i s water soluble at n e u t r a l pH a l l o w i n g
i t to be administered as true s o l u t i o n . The t o x i c i t y
of the o r i g i n a l c o l l o i d a l CLXXIV may have been due to
the I.V. a d m i n i s t r a t i o n of p a r t i c u l a t e matter.
The
a c t i v i t y of CLXXIII versus CLXXIV, against C. a l b i c a n s
i n f e c t e d mice was e s s e n t i a l l y i d e n t i c a l . The e t h y l est e r s of prostaglandins are claimed by Anderson et a l .
(473) to lead to a decrease i n diarrhea associated with
prostaglandin a d m i n i s t r a t i o n .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

PRO-DRUGS

80

The Use of Pro-Drugs to Overcome Problems of Poor


Patient Acceptance of a Product
The commercial p o t e n t i a l of a product might be
judged i n terms of a) i t s a b i l i t y to cure or suppress
some diseased s t a t e more s u c c e s s f u l l y than agents curr e n t l y a v a i l a b l e , and b) the p o t e n t i a l market produced
by that disease. However, a c r i t i c a l c o n s i d e r a t i o n often overlooked i s a p a t i e n t acceptance f a c t o r . P a i n f u l
i n j e c t i o n s might be a deterrent to the use of a drug,
e s p e c i a l l y i f m u l t i p l e dosing i s r e q u i r e d . An a n t i b i o t i c might be extremely b i t t e r t a s t i n g making i t s u i t able f o r dosing i n a capsule or coated t a b l e t but uns u i t a b l e f o r a p e d i a t r i suspensio
chewabl t a b l e
dosage form. Poor p a t i e n
p e c i a l l y with p e d i a t r i c and p o s s i b l y g e r i a t r i c prod
u c t s , can often be a deterrent f a c t o r f o r commercializ a t i o n of a product f o r that segment of the population.
Chloramphenicol (XCLV) i s an example of a u s e f u l
drug which, although s p a r i n g l y water s o l u b l e , had an
unpleasant b i t t e r t a s t e . Various d e r i v a t i v e s of XCLV
such as i t s palmitate e s t e r (CLXXV) were synthesized
and t e s t e d f o r t a s t e acceptance and t h e i r a b i l i t y to
d e l i v e r XCLV (474-476) on o r a l dosing.
Other s p a r i n g l y
soluble XCLV d e r i v a t i v e s have been synthesized i n an
e f f o r t to overcome the t a s t e problem (477-479). Because of the s p a r i n g l y water soluble nature of CLXXV,
i t presented a number of o r a l b i o a v a i l a b i l i t y problems
that were overcome by the use of a metastable polymorph
of CLXXV and the c a r e f u l screening of p a r t i c l e s i z e e f f e c t s (480). Glazko et a l . (475,481) i n t h e i r studies
of the b i o a v a i l a b i l i t y of XCLV from CLXXV noted some
unusual r e s u l t s . I t might seem that CLXXV should be
p a r t i a l l y absorbed as such and be converted to XCLV
both i n the GI t r a c t and plasma but no CLXXV was found
i n plasma. Recently Andersgaard et_ a l . (482) noted
that the d i s s o l u t i o n of CLXXV from p a r t i c l e s was c a t a lyzed by pancreatic l i p a s e and that on d i s s o l u t i o n XCLV
was r e l e a s e d . S u r p r i s i n g l y the pancreatic l i p a s e d i d
not catalyze the conversion of CLXXV to XCLV i n the
bulk phase s o l u t i o n . The authors postulate that the
pancreatic l i p a s e i s a c t u a l l y adsorbed onto the CLXXV
p a r t i c l e and a c t u a l l y catalyzes a s o l i d s t a t e hydrolys i s of the ester to i t s corresponding a c i d and a l c o h o l
fractions.
A question that probably should be asked i s why
does a drug t a s t e b i t t e r ? B i t t e r n e s s r e s u l t s from a
compound d i s s o l v i n g i n the s a l i v a of the mouth and i n t e r a c t i n g with a b i t t e r t a s t e receptor.
Molecular
t h e o r i e s on the cause of sweet and b i t t e r t a s t e have

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA

Overview and Definition

81

appeared i n the l i t e r a t u r e and w i l l not be discussed


here (483-485). The general method of overcoming a bitter taste
i n a pharmaceutical has i n v o l v e d e i t h e r a f o r m u l a t i o n
technique, i . e . coated t a b l e t , use of capsules, or prep a r a t i o n of a b i o r e v e r s i b l e , l e s s water s o l u b l e d e r i v a t i v e of the drug. CLXXV was one example of the l a t t e r
technique. The lowering of the water s o l u b i l i t y does
not block the drug-receptor i n t e r a c t i o n but simply prevents the drug from ever reaching the r e c e p t o r . I f the
molecular theory of sweet and b i t t e r receptors i s accepted, simply b l o c k i n g the e l e c t r o p h i l i c or nucleop h i l i c s i t e of i n t e r a c t i o n should s u f f i c e .
Other examples of pro-drugs used to overcome t a s t e
problems are the p a l m i t a t
discussed f u r t h e r b
s a l t of erythromycin (486), the e t h y l s u c c i n a t e and
e t h y l carbonate e s t e r s of erythromycin ( 1 7 8 , 1 7 9 ) , phosphate and carbonate e s t e r s of lincomycin ( 4 8 7 - 4 8 9 ) ,
a c y l e s t e r N-oxide oleandomycin ( 4 9 0 ) , and the t r i a c e t y l e s t e r of oleandomycin which was l e s s s o l u b l e and
therefore l e s s b i t t e r than oleandomycin ( 1 6 8 , 1 7 2 - 1 7 3 ) .
N - A c e t y l s u l f i s o x a z o l e (Lipogantrisin) and N - a c e t y l sulfamethoxypyridazine (Kynex) were two t a s t e l e s s der i v a t i v e s of s u l f i s o x a z o l e and sulfamethoxypyridazine
s u i t a b l e as p e d i a t r i c suspensions ( 4 9 1 ) . The 3 , 4 , 5 trimethoxybenzoate s a l t of t e t r a c y c l i n e was a l s o found
f

to be t a s t e l e s s

(492).

The unpleasant t a s t e of acetaminophen (CLXXVI) has


prevented i t s use i n a chewable t a b l e t f o r m u l a t i o n f o r
p e d i a t r i c p a t i e n t s . Repta and Hack ( 4 9 3 ) prepared 2 (p-acetaminophenoxy)tetrahydropyran
(CLXXVII), a prodrug of CLXXVI, which was shown to have a lower water
s o l u b i l i t y than CLXXVI and i s r e a d i l y converted to
CLXXVI under a c i d i c c o n d i t i o n s . A s e r i e s of 0 - a c y l and
O-carbonate e s t e r s of CLXXVI as pro-drugs of CLXXVI
were evaluated by D i t t e r t et a l . ( 4 9 4 - 4 9 8 ) . Their obj e c t i v e d i d not appear to be the blockage of CLXXVI's
t a s t e problem. C h l o r a l hydrate (CLXXVIII), a hypnotic,
was l i m i t e d i n i t s use by an unpleasant, b i t t e r t a s t e
and odor as w e l l as f o r m u l a t i o n problems (to be d i s cussed f u r t h e r i n the next s e c t i o n ) . Various d e r i v a t i v e s of CLXXVIII have been attempted to overcome the
t a s t e and odor problems. One example was d i c h l o r a l phenazone (CLXXIX), a molecular complex between
CLXXVIII and phenazone ( 4 9 9 ) . A l t e r n a t i v e l y CLXXVIII
i s considered to be a precursor (a pro-drug) of t r i chloroethanol (CLXXX), a high b o i l i n g point c o r r o s i v e
l i q u i d ( 5 0 0 ) . S o l i d pro-drugs of CLXXX have been promoted ( 4 9 7 , 4 9 8 , 5 0 1 , 5 0 2 ) , i n c l u d i n g t r i c l o r p h o s (CLXXXI),

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

82

PRO-DRUGS

CH CHN-^

0 <^

^)

(CLXXVII)
+

H /H 0
2

(CLXXVI)

b i s - t r i c h l o r o e t h y l carbonate (CLXXXII) and t r i c h l o r o ethyl-4-acetamidophenyl carbonate (CLXXXIII).


B i t t e r n e s s associated with amine drugs are w e l l
documented (361,362). Borodkin et_ a l . (361,362) prepared a s e r i e s of s l i g h t l y soluble ion exchange rsintes of a s e r i e s of amine drugs with the o b j e c t i v e of
preparing products s u i t a b l e f o r a chewable t a b l e t dosage form. The s l i g h t l y water soluble pyruvium pamoate
(Povan) i s described as a pleasant t a s t i n g suspension
(503) while the t r a n q u i l i z e r / a n t i h i s t a m i n e hydroxyzine
pamoate (Vistarl) and the antiemetic diphenidol
pamoate (Vontrol) are a l s o s l i g h t l y water soluble
t a s t e l e s s suspensions (504,505). S i m i l a r l y the spari n g l y water soluble napsylate s a l t of propoxyphene
(Darvon-N) was promoted as a t a s t e l e s s and stable der i v a t i v e of propoxyphene (506,508). Propoxyphene could
not be prepared i n a l i q u i d dosage form because of
s t a b i l i t y problems or i n a p e d i a t r i c dosage form because of b i t t e r n e s s . Various s u l f o n i c a c i d s a l t s were
found to be t a s t e l e s s (506,509) because of t h e i r low
aqueous s o l u b i l i t y .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

83

Overview and Definition

9H
C1CCH
3

C1_CCH0H

OH
(CLXXVIII)

(CLXXX)

ClgCCHgO Na

Cl CCH OCOCH CCl


3

OH
(CLXXXI)

CH CHNF
3

(CLXXXII)

yOCOCH CCl
2

(CLXXXIII)
The Use of Pro-Drugs to Promote S i t e S p e c i f i c D e l i v e r y
of a Drug
I d e a l l y , pro-drugs might be u s e f u l i n promoting
s i t e s p e c i f i c i t y f o r a given drug by l o c a l i z i n g the
drug i n a target organ by u t i l i z i n g e i t h e r some s p e c i f
i c p h y s i c a l or chemical property of the s i t e . For ex
ample, tumor c e l l s are postulated to contain a higher
concentration of phosphatase and amidase enzymes than
normal c e l l s (510). Therefore, i f a c y t o t o x i c drug i s
phosphorylated (assuming a s u i t a b l e f u n c t i o n a l group
f o r attachment i s a v a i l a b l e ) , the tumor c e l l s w i l l pro
v i d e a " s i n k " f o r the drug thus promoting a somewhat
s p e c i f i c accumulation of the drug at that s i t e .
Phosp h o r y l d e r i v a t i v e s of c y t o t o x i c agents such as d i e t h y l s t i l b e s t e r o l (CLXXXIVa and CLXXXIVb) and e s t r a d i o l
(CVI) have been found u s e f u l i n the treatment of pro
s t a t i c carcinoma (299,389-397) presumably due to t h i s
s p e c i f i c i t y . Phosphate esters of cytosine arabinoside
(511,512) and adenine arabinoside (43,44) have not
proven to be any more a c t i v e than the parent compounds.
The dicarbamate of CLXXXIVa, (CLXXXV), has a l s o been
suggested f o r the treatment of p r o s t a t i c carcinoma
(513). The parent compounds i n each of these cases are

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

84

PRO-DRUGS

RO

OR
(CLXXXIVa):
( CLXXXIVb): R
(CLXXXV): R
the a c t i v e agent with the precursors having l i t t l e i n trinsic activity.
L o c a l i z a t i o n of drugs at a s i t e has received some
success with the bowel s t e r i l a n t s , s u c c i n y l and phthal o y l s u l p h a t h i a z o l e (514). Both these d e r i v a t i v e s are
monoamides of sulphathiazole which, due t o t h e i r p o l a r i t y , are not w e l l absorbed from the GI t r a c t . I n the
lower i n t e s t i n e and colon, both r e l e a s e s u l p h a t h i a z o l e
which then acts as the bowel s t e r i l a n t . Another prodrug which has shown some s i t e s p e c i f i c i t y i s oxyphenis a t i n and i t s d i a c e t a t e d e r i v a t i v e (CLXXXVII). CLXXXVI
OR

(CLXXXVI): R = -H
(CLXXXVII): R = -COCH

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

85

i t s e l f i s a c t i v e as a bowel vacuant i f administered


r e c t a l l y as a s o l u t i o n . CLXXXVII i s a c t i v e o r a l l y and
i s metabolized t o CLXXXVI i n the i n t e s t i n e s and exerts
i t s vacuant p r o p e r t i e s as CLXXXVI. Bruzzese e^t a l .
(515) and Hubacher et_ a l . (516.) have surveyed the e f f e c t of a c e t y l a t i o n of d i p h e n o l i c l a x a t i v e s and found
the d i a c e t y l d e r i v a t i v e s t o be l e s s potent than the d i hydroxy metabolite per se. However, the a c e t y l a t e d der i v a t i v e s are l e s s i r r i t a t i n g and more s t a b l e when administered o r a l l y .
Other examples of s i t e s p e c i f i c i t y through prodrugs were discussed e a r l i e r , e.g., promotion of passage through the blood b r a i n b a r r i e r and changing the
permeability characteristic
variou
chelatin
agents. Methenamin
o f f e r s s i t e s p e c i f i c i t y f o r formaldehyde t o the u r i n a r y
t r a c t . When administered o r a l l y i n an e n t e r i c coated
t a b l e t , I i s absorbed and excreted i n the u r i n e . A c i d i f i c a t i o n of the u r i n e by e i t h e r d i e t a r y r e g u l a t i o n or
c o a d m i n i s t r a t i o n of a c i d i f y i n g agents such as ammonium
c h l o r i d e or sodium biphosphate promotes formation of
the n o n s p e c i f i c a n t i b a c t e r i a l formaldehyde (22). The
e n t e r i c coating of methenamine t a b l e t s i s necessary t o
prevent g a s t r i c a c i d i t y from converting I t o formaldehyde prematurely (22).
An example of a product which may be considered a
pro-drug i s selenium s u l f i d e .
Selenium d e r i v a t i v e s are
u s e f u l as a n t i s e b o r r h e i c and a n t i b a c t e r i a l agents
(517). However, the more water soluble d e r i v a t i v e s are
a l s o t o x i c due t o systemic selenium a b s o r p t i o n . S e l e nium s u l f i d e i s a very s l i g h t l y soluble selenium d e r i v a t i v e u s e f u l f o r l o c a l a p p l i c a t i o n i n the treatment of
dandruff. The poor s o l u b i l i t y allows f o r l o c a l e f f e c t
while preventing systemic t o x i c i t y .
The Use of Pro-Drugs t o E l i m i n a t e S t a b i l i t y and Other
Formulation Problems
The s t a b i l i t y of a drug i n i t s dosage form, whether a l i q u i d or s o l i d dosage form, can l i m i t the commerc i a l p o t e n t i a l of a drug product. Most drug s t a b i l i t y
problems can g e n e r a l l y be overcome v i a p h y s i c a l means
r a t h e r than chemical means. That i s , i f a drug, u s e f u l
as an i n j e c t a b l e , i s s p a r i n g l y stable i n aqueous s o l u t i o n , l y p o p h y l i z a t i o n of the drug and simple r e c o n s t i t u t i o n w i t h a solvent before a d m i n i s t r a t i o n to the pat i e n t might be the answer t o the s t a b i l i t y problem.
The pro-drug approach has been u t i l i z e d w i t h v a r y ing degrees of success as an a l t e r n a t i v e means of product s t a b i l i z a t i o n . P e n i c i l l i n s are r a t h e r unstable i n

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

PRO-DRUGS

86

aqueous s o l u t i o n due to -lactam r i n g h y d r o l y s i s zo the


corresponding p e n i c i l l o i c a c i d . Under a c i d i c condi
t i o n s the h y p e r a l l e r g e n i c , p e n i c i l l e n i c a c i d can a l s o
be formed. I n j e c t a b l e and o r a l suspension forms of
P e n i c i l l i n G (CLXXXVIII) were l i m i t e d due to t h i s i n
s t a b i l i t y . P e n i c i l l i n G degradation occurs i n s o l u
t i o n . I f a s p a r i n g l y soluble s a l t of the p e n i c i l l i n i s
employed, then the degradation becomes zero order,
since the concentration of p e n i c i l l i n i n the s o l u t i o n
remains small and constant due to replenishment of the
degraded p e n i c i l l i n from the suspension. Therefore,
the r a t e of degradation i s a f u n c t i o n of the amount of
d i s s o l v e d p e n i c i l l i n which, i n t u r n , i s a f u n c t i o n of
the s o l u b i l i t y product of the s a l t
The benzathine
procaine, and hydrabamin
(CLXXXIX, XCC and XCC r e s p e c t i v e l y )
s p a r i n g l y soluble s a l t s of p e n i c i l l i n s f o r both o r a l
and I.M. a d m i n i s t r a t i o n ( 7 - 1 1 ) . For P e n i c i l l i n G
CLXXXIX has an aqueous s o l u b i l i t y of 0 . 1 5 mg/ml, XCC
0 . 4 mg/ml, XCCI 0 . 0 7 5 mg/ml. These s p a r i n g l y water
soluble p e n i c i l l i n s a l t s which allowed the preparation
of l i q u i d p e n i c i l l i n dosage forms a l s o l e d to more sus
tained or prolonged release forms of p e n i c i l l i n u s e f u l
as s i n g l e dose I.M. i n j e c t i o n (7.)
As stated e a r l i e r , propoxyphene (XCCII) was unsta
b l e i n aqueous s o l u t i o n so preventing i t s use i n a pe
d i a t r i c l i q u i d dosage form. The napsylate s a l t of pro
poxyphene (Darvon-N, XCCIII) as a s p a r i n g l y soluble
s a l t of XCCII was formulated i n a p e d i a t r i c suspension
( 5 0 8 ) . XCCIII showed release c h a r a c t e r i s t i c s of
XCCII from the suspension and capsule dosage forms
very s i m i l a r to the p r e v i o u s l y used XCCII hydrochlo
r i d e . However, there d i d appear to be a s l i g h t and ex
pected prolonged release e f f e c t .
Erythromycin i s degraded very r a p i d l y under a c i d i c
conditions ( 1 6 6 ) . Nelson has shown that the b i o a v a i l
a b i l i t y of erythromycin from various erythromycin es
t e r s i s i n v e r s e l y p r o p o r t i o n a l to the aqueous s o l u b i l
i t y of the e s t e r s . That i s , the l e s s water soluble the
e s t e r , the b e t t e r the b i o a v a i l a b i l i t y ( 1 6 6 ) .
The three examples j u s t given represent one mode
of product s t a b i l i z a t i o n v i a the pro-drug approach.
S t a b i l i z a t i o n of a product through covalent chemical
m o d i f i c a t i o n by b l o c k i n g a decomposition s i t e or block
ing a f u n c t i o n a l group which f a c i l i t a t e s the decomposi
t i o n has also been t r i e d . H e t a c i l l i n , a more s t a b l e
pro-drug form of a m p i c i l l i n , w i l l be discussed l a t e r by
Dr. S i n k u l a . Epinephrine ( I X ) , the catecholamine d i s
cussed e a r l i e r , i s s u s c e p t i b l e to pH dependent oxida
t i o n ( 5 1 8 ) . Attempted s t a b i l i z a t i o n of IX by the addi-

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

Overview and Definition

STELLA

87

CH NH CH CH NH CH
2

Penicillin

(CLXXXIX)

y L

< c

Penicillin

(XCC)
CH

CH NH CH CH NH CH

CH.

CH(CH )
3

(CH ) CH
3

Penicillin

J 2

(XCCI)

t i o n o f the a n t i o x i d a n t , sodium b i s u l f i t e , was found


to catalyze a nonoxidative breakdown of IX t o a s u l
fonate
(XCCIV). This c a t a l y s i s was studied by Higuchi
and Schroeter (519) who found that the p-hydroxy group
of IX was necessary f o r the s u l f o n a t i o n r e a c t i o n t o
occur. Riegelman and Fischer (520) found that the
a d d i t i o n of b o r i c a c i d b u f f e r t o a s o l u t i o n of IX s t a
b i l i z e d IX against b i s u l f i t e a t t a c k . They postulated
and l a t e r i s o l a t e d epinephryl borate (XCCV), a s t a b i -

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

88

PRO-DRUGS

CH.
(CH ) NHCH CH

(j)CC H
2

"

C H

2 6 5
C

6 5
H

(XCCIII)

HO-^

\-CHCH NH ( CH ) fh^

HO

CHCH NH( CH )
2

n u

(XCCIV)
(XCCV)
l i z e d b i o r e v e r s i b l e d e r i v a t i v e of IX.
Ascorbic a c i d or v i t a m i n C i s very s u s c e p t i b l e to
o x i d a t i o n both i n s o l u t i o n and to some degree i n the
s o l i d s t a t e . This o x i d a t i o n w i l l take place only i f
the 2 , 3 - d i o l system of ascorbic a c i d i s f r e e , i . e . ,
not d e r i v a t i z e d . Such d e r i v a t i v e s as 2 and/or 3 - a c y l
( 5 2 1 ) , -benzoyl ( 1 2 9 - 1 3 2 ) , -phosphoryl ( 1 8 9 , 5 2 2 - 5 2 4 ) ,
and s u l f a t e d e r i v a t i v e s (525) have been shown to be
more stable i n s o l u t i o n and to provide s i m i l a r v i t a m i n
C a c t i v i t y as ascorbic a c i d i t s e l f . S i m i l a r l y , various
b i o r e v e r s i b l e d e r i v a t i v e s of hydrocortisone have been
shown to be quite s t a b l e ( 3 8 2 - 3 8 4 ) , whereas h y d r o c o r t i
sone i t s e l f i s quite s u s c e p t i b l e to degradation ( 5 . 2 6 ) .
S o l i d state degradations can a l s o be a problem
with some drugs. Highly unsaturated hydrocarbons, such
as v i t a m i n A and v i t a m i n D, are s u s c e p t i b l e to degrada
t i o n . G u i l l o r y and Higuchi (527) studied the s o l i d
s t a t e s t a b i l i t y of some v i t a m i n A d e r i v a t i v e s . They
found that the s o l i d s t a t e s t a b i l i t y was i n v e r s e l y pro
p o r t i o n a l to the melting point of the s o l i d , i . e . , the
higher the melting point of the d e r i v a t i v e the more
s t a b l e the product. However, as stated by the authors,
the higher melting point d e r i v a t i v e s a l s o had the lower
aqueous s o l u b i l i t y so the b i o a v a i l a b i l i t y of the more
s t a b l e products might present some problems.
Forlano et a l . ( 5 2 8 - 5 3 0 ) studied the e f f e c t of
a c y l a t i o n of v i t a m i n A a l c o h o l on the s t a b i l i t y of
v i t a m i n A. The ,-dimethylpalmityl d e r i v a t i v e was
found to be quite s t a b l e . The b i o l o g i c a l a v a i l a b i l i t y

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

89

of v i t a m i n A from the s t e r i c a l l y hindered esters using


cod l i v e r o i l and v i t a m i n A palmitate as c o n t r o l s , was
lower than the c o n t r o l s . The highest a c t i v i t y r e s u l t e d
from v i t a m i n A palmitate w i t h the ,-dimethylpalmitate
d e r i v a t i v e g i v i n g 70% b i o l o g i c a l a c t i v i t y r e l a t i v e t o
the palmitate d e r i v a t i v e ( 5 3 0 ) .
Thiamine, or v i t a m i n B]_, was found t o be unstable
when added t o p o l i s h e d r i c e . Higuchi and Windheuser
( 9 2 ) have shown thiamine t o be a very unstable com
pound. Various l i p i d s o l u b l e , s t a b l e thiamine pro
drugs, such as XX-XXIV, have been found u s e f u l as food
additives

(91-93).

The p h y s i c a l and chemical p r o p e r t i e s of a drug may


prevent i t s f o r m u l a t i o n
mercaptan (CoHc-SH,
the treatment of t u b e r c u l o s i s and leprosy ( 5 3 1 ) . How
ever, XCCVI has a very low b o i l i n g point of 3 5 which
creates obvious f o r m u l a t i o n problems. S i m i l a r l y , be
cause o f i t s odor and high vapor pressure, a problem of
p a t i e n t acceptance was created. Davies et_ a l . ( 5 3 2 - 5 3 3 )
overcame the problem by preparing a s e r i e s of t h i o e s t e r s , the most favorable being d i e t h y l d i t h i o l i s o p h t h a l a t e (XCCVII). XCCVII was a high b o i l i n g , r e l a t i v e 0
II
CSC H
0

.CH.
H0CH CH N^
CH.
2

CSC^
II 2 5
(XCCVII)

(XCCVIII)

^CH^
H0CH CH N
\m
2

CH CNH
3

C-0

(XCCIX)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

90

PRO-DRUGS

l y odorless l i q u i d , which was administered by enunction


and on absorption r e v e r t e d t o XCCVI and i s o p h t h a l i c
a c i d . S i m i l a r l y the l i q u i d t r i c h l o r o e t h a n o l was formu
l a t e d as e i t h e r CLXXVIII (500), CLXXXI (501), CLXXXII
(502) or CLXXIII (497-498)Another low b o i l i n g point
l i q u i d ,-dimethylaminoethanol or deanol (XCCVIII) was
formulated as i t s acetamidobenzoate s a l t (XCCIX, 534).
The f o r m u l a t i o n of formaldehyde as I , CXXX, and CXXXI
has already been d i s c u s s e d .
" A c c i d e n t a l " Pro-Drugs
Many of the examples that have been discussed i n
t h i s paper d i d not r e s u l t fro
planned
h t
the o p t i m i z i n g of dru
a c c i d e n t s . P r o n t o s i l (CC), the compound that provided
the c l u e that l e d t o the development of s u l f a n i l a m i d e
(CCI) and subsequently sulfonamide a n t i m i c r o b i a l
agents, was not a preconceived pro-drug of s u l f a n i l a
mide (535)* S i m i l a r l y , i t was not i n i t i a l l y r e a l i z e d
that oxazepam (CLVII) was a metabolite of diazepam
(405-408), that phenacetin gave r i s e t o acetaminophen
(CLXXVI) (536-538), that phenylbutazone had an a c t i v e
metabolite oxyphenbutazone (539), and that zoxazolamine
was metabolized t o chlorzoxazone which a l s o possessed
muscle r e l a x a n t p r o p e r t i e s (540-541).
At the same time there i s some doubt that some of
the examples g e n e r a l l y accepted and discussed as p r o -

NH 2
(CC)

AZO-REDUCTAZE

(CCI)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1.

STELLA

Overview and Definition

91

drugs are t r u l y pro-drugs.


The carbamates of mepheni s n and the 2 - s u b s t i t u t e d propanediols may have muscle
r e l a x a n t and anticonvulsant a c t i v i t y of t h e i r own (5^2,
543.). For example, there i s no proof t h a t adriamycin14-octanoate (CLXV) exerts i t s c y t o x i c a c t i v i t y due to
conversion to adriamycin (429) To prove that a d e r i v a t i v e exerts i t s a c t i v i t y as a r e s u l t of conversion to
the parent compound or some other metabolite i s not an
easy task.
Conclusion
As has been demonstrated i n t h i s review, the prodrug concept has produced many u s e f u l and p o t e n t i a l l y
u s e f u l drugs. I t mus
chemical m o d i f i c a t i o
dependen
mode of a d m i n i s t r a t i o n of the drug. Fluphenazine decanoate i s only u s e f u l as a long a c t i n g a n t i p s y c h o t i c
drug i f given as an I.M. i n j e c t i o n i n an o i l v e h i c l e
(318). The s o l v i n g of one problem may create another.
The dextran i r o n complex with a l l i t s advantages over
i o n i c i r o n was at one stage suspected of causing s a r comas (544). S o l v i n g one problem v i a a technique can
lead to other b e n e f i t s . Benzathine p e n i c i l l i n , which
provided a l i q u i d dosage form of p e n i c i l l i n , a l s o l e d
to a more sustained or prolonged r e l e a s e form of p e n i c i l l i n on I.M. i n j e c t i o n (7).
As the FDA i n the USA and governing agencies i n
other countries become more s t r i n g e n t w i t h new drug app l i c a t i o n s , many companies are t u r n i n g to the pro-drug
approach to both improve the e f f i c i e n c y and s a f e t y of
d e l i v e r y of new products and to help gain f u r t h e r patent coverage on o l d e r products which had shown d e f i c i e n c i e s . Whether new drug a p p l i c a t i o n s f o r pro-drugs
of some o l d e r , w e l l e s t a b l i s h e d products w i l l be e a s i e r
to o b t a i n i s u n c e r t a i n .
A problem that i s now w e l l recognized i s that many
u s e f u l drugs have been r e j e c t e d because, i n the screening process, l e s s than the i d e a l dosage form f o r the
drug was used. A r a t h e r a c t i v e drug may have been
overlooked simply because i t s poor aqueous s o l u b i l i t y
d i d not a l l o w a s u f f i c i e n t amount of the drug to be absorbed. Of course, i t would be an arduous and impract i c a l task to make pro-drugs of each and every e n t i t y
as i t appears. However, i f an agent i s suspected to be
h i g h l y a c t i v e based on some s t r u c t u r e - a c t i v i t y r e l a t i o n s h i p (SAR) or p r e l i m i n a r y t e s t i n g , but s u f f e r s from
poor s o l u b i l i t y or some other l i m i t a t i o n , the p o s s i b l e
development of pro-drugs at an e a r l y stage may provide
f o r greater success i n the screening of a c t i v e m e d i c i -

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

92

PRO-DRUGS

nal agents.
All the implications just discussed make the area
of pro-drugs an exciting and fruitful field for con
tinued study. It is an area where the pharmaceutical
chemist, with his knowledge and expertise in solubility
theory, pharmacokinetics and formulation variables, the
medicinal chemist, with his knowledge of synthesis, SAR
and metabolism, and the pharmacologist, with his know
ledge of mechanisms and sites of drug action and toxi
city can cooperate to optimize the delivery of an ac
tive drug to its site of action while minimizing toxi
city and unfavorable reactions to the drug. Many prob
lems associated with drugs can be overcome by the use
of pro-drug approach and i t is the hope of this author
that this review wil
this area.
Literature Cited
1. Albert, ., Nature (1958), 182, 421.
2. Albert, ., "Selective Toxicity", pp 57-63, John
Wiley and Sons Inc., New York, N.Y., 1964.
3. Harper, N. J., J. Med. Pharm. Chem. (1959), 1,
467.
4. Harper, N. J., Progr. Drug Res., (1962), 4, 221.
5. Kupchan, S. ., Casy, A. F., and Swintosky, J. V.,
J. Pharm. Sci. (1965), 54, 514.
6. Sinkula, . ., and Yalkowsky, S. H., "Rationale
for the Design of Biologically Reversible Drug Derivatives-Prodrugs" to be published as a review in
J. Pharm. Sci., 1975.
7. Sullivan, N. P., Symmes, A. T., Miller, H. C., and
Rhodehamel, Jr., H. W., Science (1948), 107, 169.
8. Hobby, G. L., Brown, E. and Patelski, R. ., Proc.
Soc. Exp. Biol. Med. (1948), 67, 6.
9. Szabo, J. L., Edwards, C. D., and Bruce, W. F.,
Antibiot. Chemother. (1951), 1, 499.
10. Samuel, M., Science (1947), 106, 370.
11. Swintosky, J. V., Rosen, E., Robinson, M. J.,
Chamberlain, R. E., and Guarini, J. R., J. Amer.
Pharm. Ass. Sci. Ed. (1956), 45, 37.
12. White, M. G., and Asch, M. J., N. Engl. J. Med.
(1971), 284, 1281.
13. Pruit, Jr., . ., Mancrief, J. ., and Mason,
A. D., U. S. Army Institute of Surgical Research,
(1967), Section 2, 1.
14. Beyer, . H., and Gourier, G. W., Science (1945),
101, 150.
15. Ariens, . J., Progr. Drug Res. (1966), 10, 429.
16. Ariens, . J., I l Farmaco Ed. Sci. (1969), 24, 3.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA

Overview and Definition

93

17. Ariens, . J., "Drug Design", Vol II, p. 2,


Academic Press, New York, . Y . , 1971.
18. Ariens, . J., "A Molecular Approach to the Modu
lation of Pharmacokinetics", published as a paper
in "The Physiological Equivalence of Drug Dosage
Forms", p. 23, presented by the Food and Drug
Directorate in Ottawa, Canada, 1969.
19. Bungaard, H., Dansk Tidsskrift Farm. (1971), 45,
73.
20. Sinkula, . ., "Molecular Modification: Deriva
tive Formation and Pharmaceutical Properties",
14th Annual National Industrial Pharmaceutical
Research Conference, Land O'Lakes, Wis., June,
1972.
21. Stella, V. J.,
57.
22. Notari, R. E . , J. Pharm. Sci. (1973), 62, 865.
23. Teorell, T., Dedrick, R. L., and Condliffe, P. G.,
"Pharmacology and Pharmacokinetics", Plenum Press,
New York, . Y . , 1974.
24. Perrier, D., and Gibaldi, M., J. Clin. Pharmacol.
(1974), 14, 415.
25. Gibaldi, M., Levy, G., and Weintraub, H., Clin.
Pharmacol. Ther. (1971), 12, 734.
26. Higuchi, T., and Davis, S. S., J. Pharm. Sci.
(1970), 59, 1376.
27. Hansch, C., "Quantitative Approaches to Pharma
cological Structure-Activity Relationships",
Chapt. 3, Cavallito, C. J., ed., Pergamon Press,
New York, . Y . , 1973.
28. Flynn, G., Yalkowsky, S. H., and Roseman, T. J.,
J. Pharm. Sci. (1974), 63, 479.
29. Ariens, . J., and Simonis, . ., "Drug Action:
Target Tissue, Dose-Response Relationships, and
Receptors", in "Pharmacology and Pharmacokinetics"
p. 163, Teorell, T., Dedrick, R. L., and
Condliffe, P. G., eds., Plenum Press, New York,
. Y . , 1974.
30. Perrier, D., and Gibaldi, M., J. Clin. Pharmacol.
(1972), 12, 449.
31. Rowland, ., "Effect of Some Physiologic Factors
in Bioavailability of Oral Dosage Forms, " in "Dosage Form
Design and Bioavailability," Chapter 6, Swarbrick, J., ed.,
Lea and Febiger, Philadelphia, Penn., 1973.
32. Barr, W. H., Drug Inf. Bull. (1969), 27.
33. Riegelman, S., and Sadee, W., "Which Drugs Can
and Should be Monitored Today and Tomorrow" in
"Clinical Pharmacokinetics", p. 169, Levy, G.,
ed., American Pharmaceutical Association, Washing
ton, D. C., 1974.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

94

PROD
- RUGS

34. Dollery, C. T., Davies, D. S., and Connolly, M.


E., Ann. . Y. Acad. Sci. (1971), 179, 108.
35. Hornykiewicz, O., Pharmacol. Rev. (1966), 18, 925.
36. Hornykiewicz, O., Pharmako-Psychiat. Neuro-Psychopharmakol. (1968), 1, 6.
37. Barbeau, ., Rev. Can. Biol. (1967), 26, 55.
38. Ernst, . ., Acta Physiol. Pharmacol. Neerl.
(1965), 15, 141.
39. Friedman, . ., and Everett, G. M., Adv. Pharma
col. (1964), 3, 83.
40. Curzon, G., Int. Rev. Neurobiol. (1968), 10, 323.
41. Cotzias, G. C., Papavasilious, P. S., Gellene, R.,
and Aronson, R. B., Trans. Ass. Amer. Phys. (1968),
81, 171.
42. Cotzias, G. C.
R., N. Engl. J. Med. (1966), 780, 337.
43. Godwin-Austen, R. B . , Tomlinson, . B . , Frears,
C. C., and Kok, H. W. L., Lancet (1969), 2, 165.
44. Peaston, M. J., and Bianchine, J. R., Brit. Med.
J. (1970), 1, 400.
45. Klawans, H. L., and Garvin, J. S., Dis. Nerv.
Syst. (1969), 30, 737.
46. McDowell, F . , Lee, J. E . , Swift, T., Sweet, T.,
Ogsbury, J. S., and Kessler, J. T., Ann. Intern.
Med. (1970), 72, 29.
47. Yahr, M. D., Duvoisin, R. C., Schear, M. J.,
Barrett, R. B., and Hoehn, M. M., Arch. Neurol.
(1969), 21, 343.
48. Barbeau, ., Can. Med. Ass. J. (1969), 101, 91.
49. Calne, D. B., Karoum, F . , Ruthven, C. R. J., and
Sandler, M., Brit. J. Pharmacol. (1969), 37, 57.
50. Mawdsley, C., Brit. Med. J. (1970), 1, 331.
51. Yoshida, H., Namba, J., Kaniike, ., and Imaizumi,
R., Jap. J. Pharmacol. (1963), 13, 1.
52. Shindo, H., Ann. Sankyo Res. Lab. (1972), 24, 1.
53. Kaplan, S. ., and Cotler, S., Paper 25 presented
at the 17th National Meeting of the Academy of
Pharmaceutical Sciences, New Orleans, 1974.
54. Bianchine, J. R., Calimlim, L. R., Morgan, J. P.,
Dujuvne, C. ., and Lassagna, L . , Ann. . Y. Acad.
Sci. (1971), 171, 126.
55. Imai, K., Suguira, M., Tamura, Z., Hirayama, K.,
and Narabayshi, H., Chem. Pharm. Bull. (1971), 17,
439.
56. Rivera-Calimlim, L., Duyuvne, C. ., Morgan, J.
P., Lassagna, L., and Bianchine, J. R., Brit. Med.
J. (1970), 4, 93.
57. Shindo, ., Miyakoshi, N . , and Takahashi, I . ,
Chem. Pharm. Bull. (1971), 19, 2490.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

95
1. STELLA Overview and Definition
58. Shindo, H., Miyakoshi, ., and Nakajima, E., Chem.
Pharm. Bull. (1972), 20, 966.
59 Shindo, H., Komai, T., Tanaka, K., Nakajima, E . ,
and Miyakoshi, N . , Chem. Pharm. Bull (1973), 21,
826.
60. Shindo, ., Nakajima, E . , Kawai, K., Miyakoshi,
N., and Tanaka, K., Chem. Pharm. Bull. (1973), 21,
817.
61. Lai, C. M., and Mason, W. D., J. Pharm. Sci.
(1973), 62, 510.
62. Martin, R. B., J. Phys. Chem. (1971), 75, 2657.
63. Niebergall, P. J., Schnaare, R. L., and Sugita, E.
T., J. Pharm. Sci. (1972), 6 1 , 232.
64. Fung, H. L., and Cheng L., J. Chem Ed (1974)
51, 106.
65. Riegelman, S., Strait
J. Pharm. Sci. (1962), 51, 129.
66. Edsall, J. T., Martin, R. B., and Hollingworth, B.
R., Proc. Nat. Acad. Sci. (1958), 44, 505.
67. Andn, . E . , Corrodi, D. L., Dahlstrm, ., Fuxe,
K., and Hkfelt, T., Life Sciences (1966), 5, 561.
68. Pinder, R. M., Nature (1970), 228, 358.
69. Creveling, C. R., Daly, J. W., Tokuyama, T., and
Witkop, B., Experientia (1969), 25., 26.
70. Daly, J . W., Creveling, C. R., and Witkop, B., J.
Med. Chem. (1966), 9, 273.
71. Borgman, R. J., McPhillips, J. J., Stitzel, R. E.,
and Goodman, I. J., J. Med. Chem. (1973), 16, 630.
72. Goldberg, L. I., Talley, R. C., McNay, J. L . ,
Progr. Cardiovasc. Dis. (1969), 12, 40.
73. Marchetti, G., Merlo, L., Noseda, V., G. Ital.
Cardiol. (1971), 1, 49.
74. Casagrande, C., and Ferrari, G., Il Farmaco Ed.
Sci. (1973), 28, 143.
75. Jones, P. H., Biel, J. H., Ours, C. W., Klundt, L.,
and Lenga, R. L., Paper and abstract presented at
the 165th meeting of the Amer. Chem. Soc., MEDI 9,
(1973).
76. Verbiscar, A. J., and Abood, L. G., J. Med. Chem.
(1970), 13, 1176.
77. Bjurlf, P., Carlstrm, S., and Rorsman, G., Acta
Med. Scand. (1967), 182, 273.
78. Kupchan, S. M., and Isenberg, A. C., J. Med. Chem.
(1967), 10, 960.
79. Mayer, S., Maickel, R. P., and Brodie, . B., J.
Pharmacol. Exp. Ther. (1959), 127, 205.
80. Brodie, B. B., Kurz, H., and Schanker, L. S., J.
Pharmacol. Exp. Ther. (1960), 130, 20.
81. Hansch, C., Steward, A. R., Anderson, S. ., and
Bentley, D., J. Med. Chem. (1968), 11, 1.
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

96

PROD
- RUGS

82. Bert1er, ., Falck, ., Owman, C., and


Rosengrenn, E., Pharmacol. Rev. (1966), 18, 369.
83. Cohen, S., Vzan, ., and Valette, G., Biochem.
Pharmacol. (1962), 11, 721.
84. Rindi, G., and Ventura, U . , Physiol. Rev. (1972),
52, 821.
85. Thomson, A. D., Baker, H., and Leevy, C. ., J.
Lab. Clin. Med. (1970), 76, 34.
86. Thomson, A. D., Baker, H., and Leevy, C. .,
Amer. J. Clin. Nutr. (1968), 21, 537.
87. Tomasulo, P. ., and Kater, R. M. H., Amer. J.
Clin. Nutr. (1968), 21, 1341.
88. Thomson, A. D., Prank, O., Baker, H., and Leevy,
C. M., Ann. Intern Med (1971) 74 529
89. Pincus, J . H.
(1972), 14, 87.
90. Montpetit, V. J. ., Anderman, F., Carpenter, S.,
Fawcett, J. S., Zborowska-slvis, D., and Giverson, H. R., Brain (1971), 94, 1.
91. Matsukawa, T., Yurugi, S., and Oka, Y . , Ann. N.
Y. Acad. Sci. (1962), 92, 430.
92. Windheuser, J. J., and Higuchi, T., J. Pharm.
Sci. (1962), 51, 354.
93. Kawasaki, C., Vitam. Horm. (1963), 21, 69.
94. Nogami, H., Hasegawa, J., and Riheshisa, T.,
Chem. Pharm. Bull. (1973), 21, 858.
95. Nogami, H., Hasegawa, J., and Noda, K., Chem.
Pharm. Bull. (1969), 17, 219.
96. Nogami, H., Hasegawa, J., and Noda, ., Chem.
Pharm. Bull. (1969), 17, 228.
97. Nogami, H., Hasegawa, J., and Noda, K., Chem.
Pharm. Bull. (1969), 17, 234.
98. Grode, G. ., Falb, R. D., Crowley, J. P., and
Truitt, . B . , Pharmacol. (1974), 11, 102.
99. Iwasaki, T., Vitamins (Kyoto) (1955), 9, 525.
100. Stella, V . , Unpublished results.
101. Matsubara, K., Vitamins (Kyoto) (1957), 12, 80.
102. Takenouchi, K., Aso, K., Shimizu, S., and Koba
yashi, T., Vitamins (Kyoto) (1962), 26, 245.
103. Takenouchi, K., Aso, K., Shimizu, S., and Koba
yashi, T., Vitamins (Kyoto) (1962), 26, 251.
104. Takenouchi, K., Aso, K., Shimizu, S., and Koba
yashi, T., Vitamins (Kyoto) (1962), 26, 257.
105. Takenouchi, K., Aso, K., Shimizu, S., and Koba
yashi, T., Vitamins (Kyoto) (1962), 26, 26l.
106. Takenouchi, ., Aso, K., and Nazaki, Y., Vita
mins (Kyoto) (1962), 26, 222.
107. Takenouchi, K., Aso, K., Minato, ., and Hirose,
S., Vitamins (Kyoto) (1962), 26, 241.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA

Overview and Definition

97

108. Morita, M., and Minesita, T., Vitamins (Kyoto)


(1966), 33, 61.
109. Takamizawa, ., and Harai, K., Chem. Pharm. Bull.
(1962), 10, 1102.
110. Takamizawa, ., Inazu, K., Nakanishi, S., Ito, H.,
Sato, H., Ando, ., Akahori, ., and Yamamoto,
R., Ann. Rept. Shionogi Res. Lab. (1967), 17, 59.
111. Minesita, T., Morita, ., and Iwata, T., Vita
mins (Kyoto) (1962), 25, 483.
112. Miyagawa, K., Mujamoto, T., and Murata, K., Vita
mins (Kyoto) (1962), 26, 31.
113. Suzuoki, Z., Suzuoki, T., and Kurihara, ., Vita
mins (Kyoto) (1962), 7, 118.
114. Howard, L., Wagner C.
d Schenker S. J
Nutr. (1974),
115 Levy, ., and Hewitt, R. R., Amer. J. Clin. Nutr.
(1971), 24, 401.
116. Chang, T., Lewis, J., and Glazko, A. J., Biochim.
Biophys. Acta (1967), 135, 1000.
117. Israel, Y . , Salazar, I . , and Rosenmann, E . , J.
Nutr. (1968), 96, 499.
118. Israel, Y., Valenzuela, J. E . , Salazar, I . , and
Vgarte, G., J. Nutr. (1970), 98, 222.
119. Yagi, K., Okuda, J., Dmitrovskii, . ., Honda,
R., and Matsubara, T., J. Vitaminol. (1961), 7,
276.
120. Yagi, K., Yamamoto, Y., and Okuda, J., Nature
(1961), 191, 171.
121. Shintani, S., Tanaka, F., Nakamura, ., and Sato,
M, J. Vitaminol. (1961), 7, 182.
122. Shintani, S., Tanaka, F., Nakamura, ., and Sato,
M., J. Vitaminol. (1961), 7, 122.
123. Editorial, Jap. Med. Gaz. (1965), 2, 12.
124. Isler, O., Experientia (1970), 26, 225.
125. Swern, D., Stirton, A. J., and Wells, P. ., Oil
and Soap (1943), 20, 224.
126. Kulesza, J., Gora, J., and Drygier, J., Zesz.
Nachem. Spozyw. (1967), 12, 29. Through Chem.
Abst. (1967), 67, 117165g.
127. Misaka, S., Tsuji, N . , Seo, S., and Iijima, S.,
Tokyo Ikadaiguku-zasshi (i960), 18, 1743.
128. Feldheim, W., and Czerny, M., Biochem. Z. (1959),
331, 150.
129. Imai, Y., Chem. Pharm. Bull. (1966), 14, 1045.
130. Imai, Y . , Matsumura, H., and Aramaki, Y . , Jap. J.
Pharmacol. (1967), 17, 330.
131. Nomura, H., and Sugimoto, ., Chem. Pharm. Bull.
(1966), 14, 1039.
132. Hoffman-LaRoche and Co., Ger. Pat. 701561 (1940).

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

98

PROD
- RUGS

133. Bloch, ., "The Design of Biologically Active


Nucleosides"in "Drug Design", Vol. IV, Chapt. 8,
Ariens, E. J., ed., Academic Press, New York, N.
Y. 1973.
134. Welch, A. D., Cancer Res. (1961), 21, 1475.
135. Sorm, F., Beranek, J., Smrt, J., Krupicka, J.,
and Skoda, J., Collection Czech. Chem. Commun.
(1962), 27, 575.
136. Chladek, S., Sorm, F . , and Smrt, J., Collection
Czech. Chem. Commun. (1962), 27, 87.
137. Zemlicka, J., Beranek, J., and Smrt, J., Collec
tion Czech. Chem. Commun. (1962), 27, 2784.
138. Beranek, J., and Pitha, J., Collection Czech.
Chem. Commun (1964) 29 625
139. Ceskoslovensk
Sorm, F . ) , Belgian Pat., 639,341 (1964).
140. Grafnettorova, J., Beranek, J., Koenig, J.,
Smahel, O., and Sorm, F., Neoplasma (1966), 13,
241.
141. Turner, R. W., and Calabresi, P., J. Invest. Der
matol. (1964), 43, 551.
142. Saksena, S. ., and Chaudhury, R. R., Indian J.
Med. Res. (1969), 57, 1940.
143. Hoeksema, H., Whitfield, G. B . , and Rhuland, L.
E., Biochem. Biophys. Res. Commun. (1961), 6,
213.
144. Posternak, T., Sutherland, E. W., and Henion, W.
F., Biochim. Biophys. Acta (1962), 65, 558.
145. Haskell, T. H., and Hanessian, S., U. S. Patent
3,651,045 (1972).
146. Gerzon, K., and Kau, D., J. Med. Chem. (1967),
10, 189.
147. Levine, R. R., and Pelkin, E. W., J. Pharmacol.
Exp. Ther. (1961), 131, 319.
148. Levine, R. R., J. Pharmacol. Exp. Ther. (1960),
129, 296.
149. Levine, R. R., Weinstock, J., Zirckle, C. S., and
McClean, R., J. Pharmacol. Exp. Ther. (1961),
131, 334.
150. Streitwieser, Jr., ., Chem. Rev. (1950), 56,
667.
151. Kusnetsov, S. G., and Ioffe, D. B., J. Gen. Chem.
USSR (1961), 31, 2289.
152. Ross, S. B . , and Frden, ., Europ. J. Pharmacol.
(1970), 13, 46.
153. Ross, S. B., Johansson, J. G., Lindborg, B., and
Dahlbom, R., Acta Pharm. Suecica (1973), 10, 29.
154. Johansson, J. G., Lindborg, B . , Dahlbom, R.,
Ross, S. B . , and Akerman, . ., Acta Pharm.
Suecica (1973), 10, 199.
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA
155.
156.
157.
158.
159.
160.
161.
162.
163.
164.
165.
166.
167.
168.
169.
170.
171.
172.
173.
174.
175.
176.
177.
178.
179.

Overview and Definition

99

Ross, S. B., Sandberg, R., Akerman, . .,


Domeii, K. E . , Stening, G., and Suensson, S.,
J. Med. Chem. (1973), 16, 787.
Lindborg, B., Johansson, J. G., Dahlbom, R., and
Ross, S. B . , Acta Pharm. Suecica (1974), 11, 401.
Ross, S. B., and Akerman, S. . ., J. Pharmacol.
Exp. Ther. (1972), 182, 351.
Kirby, W. . ., and Kind, C., Ann. . Y. Acad.
Sci. (1967), 145, 291.
Kunin, C. ., Ann. . Y. Acad. Sci. (1967), 145,
282.
Daehne, W. V., Prederiksen, E., Gundersen, E . ,
Lund, F., Mrch, P., Petersen, H. J., Roholt, K.,
Tybring, L., and Godtfredsen W O., J. Med
Chem. (1970)
Klein, J . O., and Finland, M., Amer. J. Med.
Sci. (1963), 66, 544.
Modr, Z., and Dvoracek, ., Rev. Czech. Med.
(1970), 16, 84.
Loo, J. C. K., Foltz, E. L., Wallick, ., and
Kwan, K. C., Clin. Pharmacol. Ther. (1974), 16,
35.
Perrier, D., and Gibaldi, ., J. Pharm. Sci.
(1973), 62, 1486.
Chun, A. H. C., and Seitz, J. ., J. Amer. Pharm.
Ass. (1974), NS14, 407.
Nelson, E., Chem. Pharm. Bull. (1962), 10, 1099.
Smith, J. W., Dyke, R. W., and Griffith, R. S.,
J. Amer. Med. Ass. (1953), 151, 805.
Shubin, H., Dumas, K., and Sokmensuer, ., Antibiot. Ann. (1957-1958), 679.
Meyer, C. E . , and Lewis, C., "Antimicrobial
Agents and Chemotherapy", American Society for
Microbiology (1963), 169.
Wagner, J. G., Can. J. Pharm. Sci. (1966), 1, 55.
Sobin, . ., English, A. R., and Celmer, W. D.,
Antibiot. Ann. (1954-1955), 827.
Celmer, W. D., Els, H., and Murray, K., Antibiot.
Ann. (1957-1958), 476.
Celmer, W. D., Antibiot. Ann. (1958-1959), 277.
English, A. R., and McBride, T. J., Antibiot.
Chemother. (1958), 8, 424.
English, A. R., and McBride, T. J., Proc. Soc.
Exp. Biol. Med. (1959), 100, 880.
Smith, I. M., and Soderstrom, W. H., J. Amer.
Med. Ass. (1959), 170, 184.
Stephens, V. C., Antibiot. Ann. (1953-1954), 514.
Murphy, H. W., Antibiot. Ann. (1953-1954), 500.
Clark, R. ., and Varner, E. L., Antibiot. Chemo
ther. (1957), 7, 487.
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

100
180.
181.
182.
183.
184.
185.
186.
187.
188.
189.
190.
191.
192.
193.
194.
195.
196.
197.
198.
199.

PROD
- RUGS
Jones, P. H., Perun, T. J., Rowley, . K., and
Baker, E. J., J. Med. Chem. (1972), 15, 631.
Martin, Y. C., Jones, P. H., Perun, T. J.,
Grundy, W. E., Bell, S., Bower, R. R., and Shipkowitz, N. L . , J. Med. Chem. (1972), 15, 635.
Tardew, P. L . , Mao, J. C. H., and Kennedy, D.,
Appl. Microbiol. (1969), 18, 159.
Fletcher, H. P., Murray, . ., and Weddon, T.
E.,
J.
Pharm. Sci. (1968), 57, 2101.
Morozowich, W., Sinkula, . ., MacKellar, F. .,
and Lewis, C., J. Pharm. Sci. (1973), 62, 1102.
Sinkula, . ., and Lewis, C., J. Pharm. Sci.
(1973), 62, 1757.
Mlek, P., Kolc
Antibiot. Ann
Foreman, H., "Metal Binding in Medicine", p. 82,
Seven, M. J., ed., Lippincott, Philadelphia,
Penn., 1960.
Catsch, ., Fed. Proc. (1961), 20, part II, 206.
Imai, Y . , Usui, T., Matsuzaki, T., Yokotani, H.,
Mima, H., and Aramaki, Y . , Jap. J. Pharmacol.
(1967), 17, 317.
"American Hospital Formulary Service", American
Society Hospital Pharmacists, Washington, D. C.
(1974), 84, 04.16.
Perrin, D. D., "Dissociation Constants of Organic
Bases in Aqueous Solution", p. 119, Butterworth
Co., London, England, 1965.
Barry, B. W., and Woodford, R., Brit. J. Derma
t o l . (1974), 91, 323.
"American Hospital Formulary Service", American
Society Hospital Pharmacists, Washington, D. C.
(1974), 84, 06.
Poulsen, B . , "Design of Topical Drug Products:
Biopharmaceutics", in "Drug Design", Vol. IV,
Chapt. 5, Ariens, E. J., ed., Academic Press,
New York, . Y . , 1973.
Maistrello, I., Rigamonti, G., Frova, C., and de
Ruggieri, P., J. Pharm. Sci. (1973), 62, 1455.
Fed. Regist. (1974), 39, 2471.
Wagner, J. G., Christensen, M., Sakmar, E . , Blair,
D., Yates, J. D., Willis, III, P. W., Sedman, A.
J., and Stoll, R. G., J. Amer. Med. Ass. (1973),
224, 199.
Skelly, J., and Knapp, G., J. Amer. Med. Ass.
(1973), 224, 243.
Lindenbaum, J., Mellow, M. G., Blackstone, M. D.,
and Butler, P., N. Engl. J. Med. (1971), 285,
1344.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA

Overview and Definition

101

200. V i t t i , T. G., Banes, D., and Byers, T. ., N.


Engl. J. Med. (1971), 285, 1433.
201. Higuchi, T., and Ikeda, ., J. Pharm. Sci. (1974),
63, 809.
202. Noyes, . ., and Whitney, W. R., J. Amer. Chem.
Soc. (1897), 19, 930.
203. Rietbrock, N . , Abshagen, U., Bergmann, . V . , and
Kewitz, H., Naunyn Schmiedebergs Arch. Pharmacol.
(1972), 274, 171.
204. Rietbrock, N . , Rennekamp, C., Rennekamp, H.,
Bergmann, . V . , and Abshagen, U., Naunyn
Schmiedebergs Arch. Pharmacol. (1972), 272, 450.
205. Rennekamp, H., Rennekamp, C., Abshagen, U.,
Bergmann, . V .
d Rietbrock N . Nauny
Schmiedebergs
206. Stoll, ., and Kreis, W., Schweiz. Med. Wschr.
(1953), 83, 266.
207. Rothlin, E . , Bircher, R., and Schalch, W. R.,
Schweiz. Med. Wschr. (1953), 83, 267.
208. Haberland, G., Arzneim. Porsch. (1965), 15, 481.
209. Greef, K., Schwarzmann, D., and Waschulzik, G.,
Arzneim. Porsch. (1965), 15, 483.
210. Benthe, H. F., Arzneim. Forsch. (1965), 15, 486.
211. Frster, W., and Schulzeck, S., Biochem. Pharmacol. (1968), 17, 489.
212. Cloetta, ., Arch. Exp. Path. Pharmak. (1926),
112, 261.
213. Fiehring, ., Knappe, J., and Sundermann, .,
Dt. Ges. Wesen. (1964), 19, 2439.
214. Georges, ., Pape, J., and Duvernay, G., Arch.
Int. Pharmacodyn. Ther. (1966), 164, 47.
215. Ishaq, K. S., and Gisvold, O., J. Pharm. Sci.
(1970), 59, 412.
216. Megges, S. R., and Repke, K., Naunyn Schmiede
bergs Arch. Exp. Path. Pharmak. (1961), 241, 534.
217. White, W. F., and Gisvold, O., J. Amer. Pharm.
Ass. Sci. Ed. (1952), 41, 42.
218. Hussain, ., and Rytting, J. H., J. Pharm. Sci.
(1974), 63, 798.
219. Atkinson, R. M., Bedford, C., Child, K. J., and
Tomich, E. G., Antibiot. Chemother. (1962), 12,
232.
220. Crounze, R. G., J. Invest. Dermatol. (1961), 37,
529.
221. McNall, E. G., Antibiot. Ann. (1959-1960), 7,
674.
222. Gonzalez-Ochoa, ., and Ahumada - Padilla, .,
Arch. Dermatol, (1960), 8 1 , 833.
223. Sharpe, . ., and Tomich, E. G., Toxicol. Appl.
Pharmacol. (1960), 2, 44.
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

102
PROD
- RUGS
224. Bedford, C., Busfield, D., Child, K. J., Mac
Gregor, I., Sutherland, P., and Tomich, E. G.,
Arch. Dermatol. (1960), 8 1 , 735.
225. Chiou, W. L., and Riegelman, S., J . Pharm. Sci.
(1971), 60, 1376.
226. Carrigan, P. J., and Bates, T. R., J . Pharm. Sci.
(1973), 62, 1476.
227. Fischer, L. J., and Riegelman, S., J . Pharm. Sci.
(1967), 56, 469.
228. Williams, R. T., "Detoxification Mechanisms", p.
166, John Wiley and Sons, Inc., New York, . .,
1959.
229. Magee, W. ., Armour, S. B., and Miller, O. V . ,
Biochim. Biophys Acta (1973) 306 270
230. Karim, S. M.
Gynaec. Commun
231. Schedl, H. P., and Clifton, J . ., Gastroenterol.
(1961), 41, 491.
232. Tanabe, ., and Bigley, B . , J. Amer. Chem. Soc.
(1961), 83, 756.
233. Fried, J., Borman, ., Kessler, W. ., Grabowich,
P., and Sabo, E. F . , J . Amer. Chem. Soc. (1958),
80, 2338.
234. Gardi, R., V i t a l i , R., and Ercoli, ., J . Org.
Chem. (1962), 27, 668.
235. Heller, U. S. Patent 3,608,076 (1971), through
Wilbur, R. D. (to American Cyanamid Co.), U. S.
Patent 3,772,435 (1973).
236. Anderson, ., Haymaker, W., and Henderson, .,
J . Amer. Med. Ass. (1940), 115, 2167.
237. Gardi, R., V i t a l i , R., Falconi, G., and Ercoli,
., J . Med. Chem. (1973), l6, 123.
238. Zaks, ., Jones, T., Fink, ., and Freedman, A.
M., J . Amer. Med. Ass. (1971), 215, 2108.
239. Dayton, P. B . , and Blumberg, H., Fed. Proc. Fed.
Amer. Soc. Exp. Biol. (1970), 29, 686.
240. Linder, C., and Fishman, J., J . Med. Chem. (1973),
16, 553.
241. Editorial, Chem. Eng. News (1972), 14.
242. Way, E. L., and Adler, T. K., Pharmacol. Rev.
(1960), 12, 383.
243. Kupchan, S. ., and Casy, A. F., J . Med. Chem.
(1967), 10, 959.
244. Yoshimura, ., Oguri, ., and Tsukamoto, .,
Biochem. Pharmacol. (1969), 18, 279.
245. Oguri, K., Ida, S., Yoshimura, H., and Tsukamoto,
H., Chem. Pharm. Bull. (1970), 18, 2414.
246. Mori, ., Oguri, ., Yoshimura, H., Shimomura, .,
Kamata, O., and Ueki, S., Life Sciences (1972),
11, 525.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA
247.
248.
249.
250.
251.
252.
253.
254.
255.
256.
257.
258.
259.
260.
261.
262.
263.
264.
265.
266.
267.

Overview and Definition

103

Field, L., and Sweetman, B. J., J. Org. Chem.


(1969), 34, 1799.
Field, L., Sweetman, B. J., and Bellas, ., J .
Org. Chem. (1969), 12, 62.
Sweetman, B. J., Bellas, ., and Field, L., J.
Med. Chem. (1969), 12, 888.
Srivastava, P. K., and Field, L . , J. Med. Chem.
(1973), 16, 428.
Hartles, R. L., and Williams, R. T., Biochem. J.
(1949), 44, 335.
Siuda, J . F . , and Cihonski, C. D., J. Pharm. Sci.
(1972), 61, 1856.
Wagner, J . G., Holmes, T. D., Wilkinson, P. .,
Blair, D. C., d Stoll R G. Amer Rev Resp
Dis. (1973), 108
Gibson, M. O. J., and Nagley, M. M., Tubercle
(1955), 363 209.
Jeker, K., Lauewer, H., Regli, J., and Friedrich,
T., Amer. Rev. Tub. Pul. Dis. (1959), 79, 351.
Chiou, C. ., Biochem. Pharmacol. (1971), 20,
2401.
Stempel, E . , "Dispensing of Medication", pp.
1002-1042, Martin, E . , ed., Mack Pub. Co.,
Easton, Penn., 1971.
Gray, G. D., Nichol, F. R., Mickelson, . .,
Camiener, G. W., Gish, D. T., Kelly, R. C.,
Wechter, W. J. Moxley, T. E . , and Neil, G. L . ,
Biochem. Pharmacol. (1972), 21, 465.
Warner, D. T., Neil, G. L., Taylor, A. J., and
Wechter, W. J., J. Med. Chem. (1972), 15, 790.
Neil, G. L., Buskirk, H. H., Moxley, T. E . ,
Manak, R. C., Kuentzel, S. L., and Bhuyan, . K.,
Biochem. Pharmacol. (1971), 20, 3295.
Gish, D. T., Kelly, R. C., Camiener, G.W.,and
Wechter, W. J., J. Med. Chem. (1971), 14, 1159.
Neil, G. L., Wiley, P. F., Manak, R.C.,and
Moxley, T. E . , Cancer Res. (1970), 30, 1047.
Tanaka, T., Kobayashi, H., Okumura, K., Muranishi,
S., and Sezaki, H., Chem. Pharm. Bull. (1974),
22, 1275.
James, K. C., Nicholls, P. J., and Roberts, .,
J . Pharm. Pharmacol. (1969), 21, 24.
Miescher, K., Wettstein, ., and Tschopp, E . ,
Biochem. J . (1936), 30, 1977.
Dorfman, R. I., and Shipley, R. ., "Androgens",
pp. 119-120, John Wiley and Sons, Inc., New York,
. Y . , 1956.
Junkman, K., and Witzel, ., Z. Vitamin-, Hormon-Fermentforsch. (1957), 9, 222.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

^04

PROD
- RUGS

268.

Camerino, ., and Sala, G., Progr. Drug Res.


(I960), 2, 71.
269. Diczfalusy, E . , Acta Endocrinol. (1960), 35, 59.
270. Dorner, G., and Shubert, ., Acta Biol. Med. Ger.
(1963), Suppl. II, 209.
271. Diczfalusy, E . , Perno, O., Fex, H., and Hogberg,
B., Acta Chem. Scand. (1963), 17, 2536.
272. Dirscherl, W., and Kruskemper, H. L., Biochem.
Ztschr. (1953), 323, 520.
273. Dekanski, J., and Chapman, R. N . , Brit. J . Phar
macol. (1953), 8, 271.
274. Kupchan, M., Casy, A. F . , and Swintosky, J. V.,
J. Pharm. Sci (1965) 54 514
275. Lloyd, C. W.
crinol. (1951), 11, 724.
276. Ott, A. C., Kuizenga, M. H., Lyster, S. C., and
Johnson, . ., J . Clin. Endocrinol. (1952), 12,
15.
277. Voss, . E . , Arzneim. Forsch. (1955), 5, 208.
278. Haack, E . , Stoeck, G., and Voigt, H., Arzneim.
Forsch. (1955), 5, 211.
279. Deanesly, R., and Parkes, A. S., Biochem. J .
(1936), 30, 291.
280. Schenk, ., and Junkman, ., Arch. Exp. Path.
Pharmak. (1955), 227, 210.
281. Dirscherl, W., and Dardenne, U . , Biochem. Zschr.
(1954), 325, 195.
282. Emmens, C. W., Endocrinol. (1941), 28, 633.
283. Kupperman, H. S., Aronson, S. G., Gagliani, J.,
Parsonnet, ., Roberts, M., Silver, B . , and Postiglioni, R., Acta Endocrinol. (1954), 16, 101.
284. Meier, R., and Tschopp, E . , Naunyn Schmeidebergs,
Arch. Exp. Path. Pharmak. (1955), 226, 532.
285. Gould, D., Finckenor, L., Herschberg, . B . ,
Pearlman, P., Cassidy, J., Margolin, S., and
Spoerlein, M. T., Chem. Ind. (1955), 1424.
286. Baggett, B . , Engel, L. L., Savard, K., and Dorfman, R. I., J . Biol. Chem. (1956), 221, 931.
287. Ruzicka, L . , and Kagi, ., Helv. Chim. Acta
(1936), 19, 842.
288. Sulman, F. G., Horokeach Haivari (1958), 7, 76.
289. Kishimoto, Y . , Arch. Biochem. Biophys. (1973),
159, 528.
290. McEwen, B. S., Pfaff, D. W., and Zigmond, R. E . ,
Brain Res. (1970), 21, 17.
291. Pala, G., Casadio, S., Mantegani, ., Bonardi,
G., and Coppi, G., J. Med. Chem. (1972), 15, 995.
292. Rapala, R. T., Kraay, R. J., and Gerzon, ., J .
Med. Chem. (1965), 8, 580.
293. van der Vies, J., Acta Endocrinol. (1965), 49, 271,
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA

Overview and Definition

105

294. Chaudry, M. A. Q., and James, K. C., J. Med.


Chem. (1974), 17, 157.
295. DeVisser, J., and Overbeck, G. ., Acta Endocrin
o l . (1960), 35, 405.
296. "American Hospital Formulary Service", American
Society of Hospital Pharmacists, Washington, D.C.
(1974), 68, 08.
297. Seay, D. G., Bradshaw, J. D., and Nicol, N. T.,
Cancer Chemother. Rep. (1972), 56, 89.
298. Kuhl, H., and Taubert, H. D., Steroids (1973),
22, 73.
299. Diczfalusy, E . , and Westman, ., Acta Endocrinol.
(1956), 21, 321.
300. Falconi, G., Galletti F . Celasco G.
d
Gardi, R., Steroids
301. Ferrin, J., J. Clin. Endocrinol. (1952), 12, 28.
302. Gardi, R., Vitali, R., Falconi, G., and Erloli,
., J . Med. Chem. (1973), 16, 123.
303. Parkes, A. S., J. Endocrinol. (1943), 3, 288.
304. Parkes, A. S., Biochem. J . (1937), 31, 579.
305. Freed, S. C., Eisin, W. ., and Greenhill, J . P.,
J . Amer. Med. Ass. (1942), 119, 1412.
306. Ferno, O., Fex, H., Hogberg, B . , Linderot, T.,
Veige, S., and Diczfalusy, E . , Acta Chem. Scand.
(1958), 12, 1675.
307. Tillinger, K. G., and Westman, ., Acta Endocrin
o l . (1957), 25, 113.
308. American Home Products Co., U. S. Patent
3, 647, 784 (March 7, 1972).
309. American Home Products Co., U. S. Patent
3, 649, 621 (March 14, 1972).
310. Yamada, J., Toko Igakkai Zasshi (1959), 6, 481.
311. Rizkallah, T. H., and Taymor, M. L., Adv. Planned
Parenthood Int. Congr. Series (1967), #138, 111.
312. Zartman, E. R., Adv. Planned Parenthood Int.
Congr. Series (1967), #138, 116.
313. Haspels, . ., Ned. Tijdschr. Geneesk. (1970),
114, 61.
314. "American Hospital Formulary Services", American
Society of Hospital Pharmacists, Washington, D.C.
(1974), 68, 32.
315. Gross, F . , and Tschopp, E . , Experientia (1952),
8, 75.
316. "American Hospital Formulary Service", American
Society of Hospital Pharmacists, Washington, D.C.
(1974), 68, 04.
317. Winter, C. ., and Porter, C. C., J. Amer. Pharm.
Ass. Sci. Ed. (1957), 46, 515.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

106

PROD
- RUGS

318. "American Hospital Formulary Service", American


Society of Hospital Pharmacists, Washington, D.C.
(1974), 28, 16.08.
319. Enerheim, B., Gottfries, C. G., and Sunden, C.,
Nor. Psyk. Tidskr. (1970), 24, 239.
320. Ebert, A. G., and Hess, S. ., J . Pharmacol. Exp.
Ther. (1965), 148, 412.
321. Kincross-Wright, J., and Charalampous, K. D., J.
Neuropsychiat. (1965), 1, 66.
322. Mishinsky, J., Khazen, K., and Sulman, F. G.,
Neuroendocrinol. (1969), 4, 321.
323. Dreyfus, J., Ross, J r . , J. J., and Schreiber, E.
C.,
J.
Pharm. Sci. (1971), 60, 829.
324. Denham, J., and Adamson L . Act Psychiat
Scand. (1971)
325. Nymark, M., Franck, K. F . , Pedersen, V., Boeck,
V., and Nielsen, I. M., Acta Pharmacol. Toxicol.
(1973), 33, 363.
326. Jrgensen, ., Over, K. F . , and Hansen, V . , Acta
Pharmacol. Toxicol. (1971), 29, 339.
327. Villeneuve, ., Pires, ., Jus, ., Lachance, R.,
and Drolet, ., Cur. Ther. Res. (1972), 14, 696.
328. Ayd, F. J., Int. Drug Ther. Newsletter (1972), 7,
1.
329. Villeneuve, ., and Simon, P., J. Ther. (1971),
2, 3.
330. Thomsen, J. B . , and Birkerod, D., Acta Psychiat.
Scand. (1973), 49, 119.
331. Julou, L . , Bourat, G., Ducrot, R., Fournel, J.,
and Garrett, C., Acta Psychiat. Scand. (1973),
241, 9.
332. Elslager, E. F . , Tendick, F. H., and Werbel, L.
., J . Med. Chem. (1969), 12, 600.
333. Elslager, E. F . , Progr. Drug Res. (1969), 13,
170.
334. Thompson, P. E . , Int. J. Lepr. (1967), 35, 605.
335. Carrington, H. C., Crowther, A. F . , Davey, D. G.,
Levi, . ., and Rose, F. L., Nature, (1951),
168, 1080.
336. Waitz, J. ., Olszewski, B. J., and Thompson, P.
E., Science (1963), l 4 l 723.
337. Ryley, J . F . , Brit. J . Pharmacol. (1953), 8, 424.
338. Coatney, G. R., Contacos, P. G., and Lunn, J. S.,
Amer. J. Trop. Med. Hyg. (1964), 13, 383.
339. Crowther, A. F . , and Levi, . ., Brit. J . Phar
macol. (1953), 8, 93.
340. Carrington, H. C., Crowther, A. F., and Stacey,
G. J., J. Chem. Soc. (1954), 1017.
341. Elslager, E. F . , and Worth, D. F . , Nature (1965),
206, 630.
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA

Overview and Definition

107

342. Thompson, P. E . , Olszewski, B. J., Elslager, E.


F., and Worth, D. F . , Amer. J. Trop. Med. Hyg.
(1963), 12, 481.
343. Elslager, E. F . , Gavrilis, . B., Phillips, A.
., and Worth, D. F . , J. Med. Chem. (1969), 12,
357.
344. Elslager, E. F . , Capps, D. B . , and Worth, D. F . ,
J . Med. Chem. (1969), 12, 597.
345. Halpern, ., Shaftel, N . , and Bovi, A.J.M.,
Amer. J. Pharm. (1958), 130, 190.
346. Levine, R. H., Zaks, ., Fink, M., and Freedman,
A. (1972), New York Medical College and Metro
politan Hospital Center. See Ref. 24l.
347. Caldwell, H. C. Rednick A B . Scott G C.,
Yakatan, G. J.,
(1970), 59, 1689.
348. Shibini, . . ., Nasser, . ., and Motarvi,
M. M., Pharmazie (1971), 26, 630.
349. Gray, A. P., and Robinson, D. S., J. Pharm. Sci.
(1974), 63., 159.
350. Gray, A. D., and Robinson, D. S., First Inter
national Conference on Narcotic Antagonists,
Airlie House, Warrenton, Va., November 1972.
351. Cavallito, C. J., and Jewel, R., J. Amer. Pharm.
Assoc. Sci. Ed. (1958), 47, 165.
352. Thomspon, R. E . , and Hecht, R. ., Amer. J. Clin.
Nutr. (1959), 7, 311.
353. Lawrence, R. D., Brit. Med. J. (1955), 1, 603.
354. Benedetti, M. S., and Larue, D. ., Arzneim.
Forsch. (1973), 23, 826.
355. Hinsvark, O. N . , Truant, A. P., Jenden, D. J.,
and Steinborn, J . ., J. Pharmacokin. Biopharm.
(1973), 1, 319.
356. Miller, J r . , W. C., Marcotte, D. B . , and McCurdy,
L . , Curr. Ther. Res. (1973), 15, 700.
357. Garrett, . ., Clin. Med. (1956), 3, 1185.
358. Bundalo, T. S., Fugazzato, D. J., and Wyczalek,
J. V . , Amer. J. Trop. Med. Hyg. (1969), l 8 , 50.
359. Haves, H. L., and Lynch, J . E . , J. Parasitol.
(1967), 53, 1085.
360. Saias, F . , Jondet, ., and Phillipe, J., Ann.
Pharm. Fr. (1969), 27, 557.
361. Borodkin, S., and Yunker, . ., J. Pharm. Sci.
(1970) , 59, 481.
362. Borodkin, S., and Sundberg, D. P., J. Pharm. Sci.
(1971) , 70, 1523.
363. Loucas, S. P., and Haddad, . ., J. Pharm. Sci.
(1972) , 61, 985.
364. Myddleton, W. W., J. Soc. Cosmetic Chem. (1960),
11, 192.
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

108

PROD
- RUGS

365.
366.

Cohen, S., Drug Cosmetic Ind. (1957), 8 l , 306.


Weiner, . Z., and Zilkha, ., J. Med. Chem.
(1973), 16, 573.
367. Garner, D. D., and Garson, L. R., J. Pharm. Sci.
(1973), 62, 2049.
368. Greenblatt, D. J., Shader, R. I., and Koch-Wesser, J., N. Engl. J . Med. (1974), 291, 1116.
369. Gamble, J. A. S., MacKay, J. S., and Dundee, J .
W., Brit. J . Anaesth. (1973), 45, 926.
370. Gamble, J . A. S., MacKay, J . S., and Dundee, J .
W., Brit. J . Anaesth. (1973), 45 1085.
371. Baldwin, J., and Amerson, A. B . , Amer. J. Hosp.
Pharm. (1973), 30, 837.
372. Rowland, ., "Clinical Pharmacology"
27 Mac
millan Co., Ne
373. Dam, M., and Olesen, V., Neurol. (1966), 16, 288.
374. Serrano, E. E . , Roye, D. B . , and Hammer, R. H.,
Neurol. (1973), 23, 311.
375. Wilensky, A. J., and Lowden, J. ., Neurol.
(1973), 23, 318.
376. Lange, W. E . , and Stein, M. E . , J . Pharm. Sci.
(1964), 53, 435.
377. Kawamura, M., Yamamoto, R., and Fujisawa, S.,
Yakugaku Zasshi. (1971), 91, 879.
378. "The Merck Index", p. 86l, Eighth Ed., Stecher,
P. G., ed., Merck and Co., Rahway, N. J., 1968.
379. Melby, J . C., and St. Cyr, M., Metabolism (1961),
10, 75.
380. Novak, E . , Stabbs, S., and Chodos, D. J., Clin.
Pharmacol. Ther. (1972), 13., 148.
381. "The Merck Index", p. 542, Eighth Ed., Stecher,
P. G., ed., Merck and Co., Rahway, . Y., 1968.
382. Yamamoto, R., Fujisawa, S., and Kawamura, M.,
Yakugaku Zasshi (1971), 91, 855.
383. Kawamura, M., Yamamoto, R., and Fujisawa, S.,
Yakagaku Zasshi (1971), 91, 863.
384. Kawamura, M., Yamamoto, R., and Fujisawa, S.,
Yakugaku Zasshi (1971), 91, 871.
385. Flynn, G., and Lamb, D. J., J. Pharm. Sci. (1970)
59, 1433.
386. Kampman, J., Hansen, J . ., Siersbaek-Nielsen,
K., and Laursen, H., Clin. Pharmacol. Ther.
(1972), 13, 516.
387. Sandman, B . , "The Chemistry of Chloramphenicol
3-Monosuccinate", Ph.D. thesis, U. Wisconsin,
1968.
388. Sandman, B . , Szulczewski, D., Winheuser, J., and
Higuchi, T., J. Pharm. Sci. (1970), 59, 427.
389. Druckrey, H., and Raabe, S., Klin. Wschr. (1952),
30, 882.
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA

Overview and Definition

109

390. Brandes, D., and Bourne, G. H., Lancet (1955),


481.
391. Wilmanns, H., Medizinsche (1954), 1, 17.
392. Flocks, R. H., Marberger, H., Begley, B. J., and
Prendergast, L. J., J. Urol. (1955), 74, 549.
393. Gadermann, ., Klin. Wschr. (1952), 30, 882.
394. Framondino, M., Marino, G., Randazzo, G., and
Scardi, V . , Il Farmaco Ed. Sci. (1971), 26, 294.
395. Druckrey, H., Dtsch. Med. Wschr. (1952), 77,
1495.
396. Druckrey, ., Dtsch. Med. Wschr. (1952), 77,
1534.
397. Jacob, H., and Rothauge C. Zschr Urol (1956),
49, 301.
398. "The Merck Index"
cher, P. G., ed., Merck and Co, Rahway, N. J.,
1968.
399. Glazko, A. J., Carnes, H. ., Kazenko, ., Wolf,
L. M., and Reutner, T. F . , Antibiot. Ann. (19571958), 792.
400. Ross, S., Puig, J. R., and Zaremba, . ., Anti
biot. Ann. (1957-1958), 803.
401. Payne, H. M., and Hachney, J r . , R. L., Antibiot.
Ann. (1957-1958), 821.
402. McCrurab, Jr., F. R., Snyder, M. J., and Hicken,
W. J., Antibiot. Ann. (1957-1958), 837.
403. LePage, G. ., Lin, Y. T., Orth, R. E . , and Gott
lieb, J . ., Cancer Res. (1972), 32, 244l.
404. Hanessian, S., J. Med. Chem. (1973), 16, 290.
405. Kaplan, S. ., Jack, M. L., Alexander, ., and
Weinfeld, R. E . , J . Pharm. Sci. (1973), 62, 1789.
406. DeSilva, J . A. F . , Koechlin, . ., and Bader,
G., J . Pharm. Sci. (1966), 55, 692.
407. Marcucci, F . , Fanelli, R., Mussini, E . , and
Garattini, S., Europ. J. Pharmacol. (1969), 7,
307.
408. Berlin, ., Sirvers, ., Agurell, S., Hiort, .,
Sjqvist, F . , and Strm, S., Clin. Pharmacol.
Ther. (1972), 13, 733.
409. Walkenstein, S. S., Wiser, R., Gudmundsen, C. H.,
Kimmel, H. B., and Corradino, R. ., J. Pharm.
Sci. (1964), 51, 1181.
410. Salmona, M., Saronio, C., Bianchi, R., Marcucci,
F., and Mussini, E . , J. Pharm. Sci. (1974), 63,
222.
411. Pelzer, H., and Maass, D., Arzniem. Forsch.
(1969), 6, 1652.
412. Graziani, G., and DeMarchi, F . , Boll. Soc. Ital.
Biol. Sper. (1967), 43, 1422.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

110

PROD
- RUGS

413. Babbini, M., DeMarchi, F., Montanaro, N . ,


Strocchi, P., and Torrielli, M. N . , Arzneim.
Forsch. (1969), 19, 1931.
414. Stella, V . , and Higuchi, T., J . Pharm. Sci.
(1973), 62, 962.
415. Atkinson, J r . , A. J., and Davidson, R., Ann. Rev.
Med. (1974), 25, 99.
416. "The Merck Index", pp. 652-653, Eighth Ed.,
Stecher, P. G., ed. Merck and Co., Rahway, N. Y . ,
1968.
417. Greenberg, F. H., Leung, . K., and Leung, M.,
J . Chem. Ed. (1971), 48, 632.
418. "American Hospital Formulary Service", American
Society of Hospital Pharmacists Washington D
C. (1974), 88
419. Zitko, . ., Howes, J . F., Razdan, R. K.,
Dalzell, H. C., Sheehan, J . C., Pars, H. G.,
Dewey, W. L., and Harris, L. S., Science (1972),
179 442.
420. L i t t l e , A. D., U. S. Patent 3, 728, 360 (April 17,
1973).
421. Editorial, Chem. Eng. News (1974), Sept. 30,
15.
422. Marcus, ., U. S. Patent 3, 230, 143 (1966).
423. Butler, T. C., J . Pharmacol. Exp. Ther. (1952),
104, 299.
424. Bogue, J . Y . , and Carrington, H. C., Brit. J .
Pharmacol. (1953), 8, 230.
425. Kutt, H., Clin. Pharmacol. Ther. (1974), 16, 243.
426. Husman, J . W., Pharm. Weekbl. (1969), 104, 799.
427. Fujimoto, J . M., Mason, W. H., and Murphy, M.,
J . Pharmacol. Exp. Ther. (1968), 159, 379.
428. Bogan, J., and Smith, H., J. Pharm. Pharmacol.
(1968), 20, 64.
429. Arcamone, F . , Franceschi, G., Minghetti, .,
Denco, S., Redaelli, S., DiMarco, A. D., Casazza,
A. M., Dasdia, T., DiFronzo, G., Guiliani, P.,
Lenaz, L . , Necco, ., and Soranzo, C., J . Med.
Chem. (1974), 17, 335.
430. Davenport, H. W., Gastroenerol. (1965), 49, 189.
431. Davenport, H. W., Gastroenterol. (1965), 49, 238.
432. Smith, M. J . H., and Smith, P. ., "The Salicy
lates", pp. 235-257, Interscience Pub., New York,
N. Y . , 1966.
433. Stubb, L. Th. F. L., Pietersen, J . H., and Van
Heulen, C., Brit. Med. J . (1962), 675.
434. Pierson, R. N . , Holt, P. R., Watson, R. M., and
Keating, R. P., Amer. J . Med. (196l), 31, 259.
435. Menguy, R., Gastroenterol. (1966), 51, 430.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA

Overview and Definition

111

436. Wood, P. . ., Harvey-Smith, . ., and Dixon,


A. S. J., Brit. Med. J. (1962), 669.
437. Hurst, ., Brit. Med. J. (1943), 768.
438. Leonards, J . P., and Levy, G., J. Pharm. Sci.
(1970), 59, 1511.
439. Anderson, K. W., Arch. Int. Pharmacodyn. (1964),
152, 379.
440. Douthwaite, A. H., Lancet (1954), 917.
441. Davenport, H. W., Gastroenterol. (1964), 46, 245.
442. Croft, D. ., Cuddigan, J. H. P., and Sweetland,
C., Brit. Med. J. (1972), 545.
443. Misher, ., Adams, H. J., Fishier, J . J., and
Jones, R. G., J . Pharm. Sci. (1968), 57, 1128.
444. Dittert, L. W. Caldwell H C., Ellison T.
Irwin, G. M.
V., J . Pharm. Sci. (1968), 57, 828.
445. Hussain, ., Yamasaki, M., and Truelove, J . E . ,
J. Pharm. Sci. (1974), 63, 627.
446. Schering Corp., U. S. Patent 3, 767, 811 (Oct. 23,
1973).
447. Kutnowskie Zakay Farmaceutyzine Polfa, French
Patent 1, 604, 123 (Aug. 20, 1971).
448. Bailey, D. ., Wood, D., Johnson, R. E . ,
McAuliff, J . P., Bradford, J . C., and Arnold,
., J . Med. Chem. (1972), 15, 344.
449. Glamkowski, E. J., Gal, G., and Sletzinger, .,
J . Med. Chem. (1973), 16, 176.
450. Izquierdo, M., Gomis, P., U. S. Patent 3, 607, 881
(Sept. 21, 1971).
451. Ludwig, G., and Ache, I. ., Arzneim. Forsch.
(1973), 23, 1226.
452. Riedel, R., and Schoetensack, W., Arzneim.
Forsch. (1973), 23, 1215.
453. Konig, J., Knoche, C., and Schafer, ., Arzneim.
Forsch. (1973), 23, 1246.
454. Benakis, ., Tsoukas, G., and Glasson, B . ,
Arzneim. Forsch. (1973), 23, 1231.
455. Jones, W. G. ., Thorp, J . M. T., and Waring, W.
S., (to Imperial Chemical Industries Ltd.) Brit.
Patent 860, 303 (1961).
456. Barrett, . ., and Throp, J . M. T., Brit. J .
Pharmacol. Chemother. (1968), 32, 38l.
457. Klebanov, . ., Farmakol. Toksikol. (1970), 33,
324. (through Chem. Abstr. 73, 64752m).
458. Beirsdorf, A. G., West Ger. Patent 2, 212, 830
(Sept. 27, 1973).
459. Beirsdorf, A. G., West Ger. Patent 2, 212, 831
(Sept. 27, 1973).

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

112

PROD
- RUGS

460.

Novak, ., Wagner, J . G., and Lamb, D. J., Int.


J. Clin. Pharmacol., Ther. Toxicol. (1970), 3,
201.
461. Morozowich, W., Lamb, D. J., DeHaan, R. M., and
Gray, J . E . , p. 63, Abstracts of Papers, A.Ph.A.
Academy of Pharmaceutical Sciences, Washington,
D. C. Meeting, April 1970.
462. Martin, L. E . , Bates, C. M., Beresford, C. R.,
Donaldson, J . D., McDonald, F. F . , Dunlop, D.,
Sheard, P., London, E . , and Twigg, G. D., Brit.
J . Pharmacol. (1955), 10, 375.
463. Beresford, C. R., Golberg, L . , and Smith, J . P.,
Brit. J . Pharmacol. (1957), 12, 107.
464. Fletcher, F . and London E . Brit Med J.
(1954), 984.
465. Olsson, K. S., and Weinfeld, ., Acta Med. Scand.
(1972), 192, 543.
466. Olsson, K. S., Lundvall, O., and Weinfeld, .,
Acta Med. Scand. (1972), 191, 49.
467. Olsson, K. S., Acta Med. Scand. (1972), 192, 551.
468. Terato, K., Fujita, T., and Yoshino, Y . , J.
Pharm. Soc. Jap. (1972), 92, 1247.
469. Terato, K., Hiramatsu, Y . , and Yoshino, Y . , Amer.
J . Dig. Dis. (1973), 18, 129.
470. Terato, ., Fujita, T., and Yoshino, Y . , Amer. J.
Dig. Dis. (1973), 18, 121.
471. Bonner, D. P., Mechlinski, W., and Schaffner, C.
P., J . Antibiot. (1972), 25, 26l.
472. "The Merck Index", p. 75, Eighth Ed., Stecher, P.
G., ed. Merck and Co., Rahway, N. J., 1968.
473. Anderson, . H., and Weinshenker, N. M., U. S.
Patent 3, 723, 473 (March 27, 1973).
474. Edgerton, W. H., U. S. Patent 2, 662, 906 (1953).
475. Glazko, A. J., Edgerton, W. H., Dill, W. ., and
Lenz, W. R., Antibiot. Chemother. (1952), 2, 234.
476. Siegert, R., and Vmel, W., Anzneim. Forsch.
(1956), 6, 714.
477. Taylor, E. P., J. Pharm. Pharmacol. (1953), 5,
254.
478. Houtman, R. L., U. S. Patent 3, 447, 926 (May 6,
1969).
479. Sumitomo Chem. Co. Ltd., West Ger. Patent
2, 244, 179 (March 29, 1973).
480. Aguiar, A. J., Krc, J., Kinkel, A. W., and Samyn,
J . C., J. Pharm. Sci. (1967), 56, 847.
481. Glazko, A. J., Dill, W. ., Kazenko, ., Wolf, L.
., and Carnes, . ., Antibiot. Chemother.
(1958), 8, 516.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA

Overview and Definition

482. Andersgaard, ., Finholt, P., Gjermundsen, R.,


and Hyland, T., Acta Pharm. Suecica (1974), 11,
239.
483. Kier, L. B., J . Pharm. Sci. (1972), 61, 1394.
484. Boncrieff, R. W., "The Chemical Senses", pp. 490543, Leonard Hill, London, England, 1967.
485. Kubota, T., and Kubo, I . , Nature (1969), 223, 97.
486. Jones, P. ., Rowley, . K., Weiss, A. L.,
Bishop, D. L . , and Chun, A. H. C., J . Pharm. Sci.
(1969), 58, 337.
487. Morozowich, W., Sinkula, . ., MacKellar, F. .,
and Lewis, C., J . Pharm. Sci. (1973), 62, 1102.
488. Sinkula, . ., and Lewis, C., J . Pharm. Sci.
(1973) , 62, 1757
489. Morozowich, W.
MacKellar, F. ., Lewis, C., Stern, K. F., and
Rowe, E. L . , J. Pharm. Sci. (1969), 58, 1485.
490. Celmer, C. D., Canad. Patent 779, 315 Feb. 27,
1968).
491. "American Hospital Formulary Service", American
Society of Hospital Pharmacists, Washington, D.C.
(1974) , 8, 24.
492. Officina Terapeutica Italiana S r l , French Patent
2, 099, 449 (Mar. 17, 1972).
493. Repta, A. J., and Hack, J., J . Pharm. Sci.
(1973), 62, 1892.
494. Dittert, L. W., Caldwell, H. C., Adams, H. J.,
Irwin, G. ., and Swintosky, J . V . , J . Pharm.
Sci. (1968), 57, 774.
495. Dittert, L. W., Irwin, G. M., Chong, C. W., and
Swintosky, J . V., J. Pharm. Sci. (1968), 57, 780.
496. Rattle, E. S., Shami, E. G., Dittert, L. W., and
Swintosky, J . V . , J. Pharm. Sci. (1970), 59,
1738.
497. Swintosky, J. V., Caldwell, H. C., Chong, C. W.,
Dittert, L. W., and Irwin, G. M., J . Pharm. Sci.
(1968), 57, 752.
498. Swintosky, J . V . , Adams, H. J., Caldwell, H. C.,
Dittert, L. W., Ellison, T., and Rivard, D. E . ,
J. Pharm. Sci. (1966), 55, 992.
499. Rice, W. B., and McColl, J . W., J . Amer. Pharm.
Assoc. Sci. Ed. (1956), 45, 137.
500. Butler, T. C., J . Pharmacol. Exp. Ther. (1948),
92, 49.
501. Editorial, Drug Intel. Clin. Pharm. (1973), 7,
126.
502. Caldwell, H. C., Adams, H. J., Rivard, D. E . , and
Swintosky, J . V . , J. Pharm. Sci. (1967), 56, 920.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

114
503.
504.
505.
506.
507.
508.
509.
510.
511.
512.
513.
514.
515.
516.
517.
518.
519.
520.
521.
522.
523.
524.

PROD
- RUGS
"American Hospital Formulary Service", American
Society of Hospital Pharmacists, Washington, D.C.
(1974), 8, 08.
"American Hospital Formulary Service", American
Society of Hospital Pharmacists, Washington, D.C.
(1974), 28, 16.08.
"American Hospital Formulary Service", American
Society of Hospital Pharmacists, Washington, D.C.
(1974), 56, 20.
Gruber, J r . , C. ., Stephens, V. C., and T e r r i l l ,
P. ., Toxicol. Appl. Pharmacol. (1971), 19, 423.
Emmerson, J . L . , Gibson, W. R., and Anderson, R.
C., Toxicol. Appl. Pharmacol. (1971), 19, 445.
Wolen, R. L., Gruber J r . C M. Kiplinger G
F., and Scholz
(1971), 19, 480.
Stephens, V. C., U. S. Patent 3, 728, 379 (Apr. 17,
1973).
Shapira, F . , Adv. Cancer Res. (1973), 18, 77.
Wechter, W. J., J. Med. Chem. (1967), 10, 762.
Smith, C. G., Buskirk, H. H., and Lumis, W. L . ,
J . Med. Chem. (1967), 10, 774.
Fabrica de Medicamente Biofarm. West Ger. Patent
2, 231, 486 (Feb. 15, 1973).
"American Hospital Formulary Service", American
Society of Hospital Pharmacists, Washington, D.C.
(1974), 8, 24.
Bruzzese, T., Gio, ., and Riva, M., Il Farm. Ed.
Sci. (1972), 28, 121.
Hubacher, . ., and Doernberg, S., J. Pharm.
Sci. (1964), 53, 1067.
"The Merck Index", pp. 940-941, Eighth Ed.,
Stecher, P. G., ed., Merck and Co., Rahway, . Y.,
1968.
Sokoloski, T. D., and Higuchi, T., J. Pharm. Sci.
(1962), 51, 172.
Higuchi, T., and Schroeter, L. C., J. Amer. Chem.
Soc. (1960), 82, 1904.
Riegelman, S., and Fischer, . Z., J . Pharm. Sci.
(1962), 51, 206.
Nomura, ., and Sugimoto, ., Chem. Pharm. Bull.
(1966), 14, 1039.
Nomura, H., Kuwayama, M., Ishiguro, T., and Morimoto, S., Chem. Pharm. Bull. (1971), 19, 341.
Nomura, H., Shimomura, M., and Morimoto, S.,
Chem. Pharm. Bull. (1971), 19, 1433.
Cutolo, E . , and Larizza, ., Gazz. Chim. Ital.
(1961), 91, 964.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

1. STELLA

Overview and Definition

115

525. Seib, P. ., Liang, Y. T., Lee, C. H., Hoseney,


C., and Deyoe, C. W., J . Chem. Soc. Perkin I
(1974), 1220.
526. Pitman, I. H., Higuchi, T., Alton, M., and Wiley,
R., J . Pharm. Sci. (1972), 61, 918.
527. Guillory, J . K., and Higuchi, T., J . Pharm. Sci.
(1962), 51, 100.
528. Porlano, A. J., Jarowski, C. I., and Hammer, H.
F., J . Pharm. Sci. (1968), 57, 1184.
529. Forlano, A. J., Jarowski, C. I., Hammer. H. F . ,
and Merritt, E. G., J. Pharm. Sci. (1970), 59,
121.
530. Forlano, A. J., J . Pharm. Sci. (1971), 60, 6l6.
531. Davies, G. E .
d Driver G W. Natur (1958)
182, 664.
532. Davies, G. E . , Driver, G. W., Hoggarth, E . ,
Martin, A. R., Paige, M. F. C., Rose, F. L . , and
Wilson, B. R., Brit. J . Pharmacol. (1956), 11,
351.
533. Davies, G. E . , and Driver, G. W., Brit. J . Phar
macol. (1957), 12, 434.
534. "American Hospital Formulary Service", American
Society of Hospital Pharmacists, Washington, D.C.
(1974), 28, 20.
535. Trfoul, J., Trfoul, J., N i t t i , F., and Bovet,
D.
,
Comp. Rend. Soc. Biol. (1935), 120, 756.
536. Brodie, . B . , and Axelrod, J., J . Pharmacol.
Exp. Ther. (1949), 97, 58.
537. Brodie, . B . , and Axelrod, J., J . Pharmacol.
Exp. Ther. (1948), 94, 29.
538. Flinn, F. B . , and Brodie, . B . , J. Pharmacol.
Exp. Ther. (1948), 94, 76.
539. Burns, J . J., Rose, R. K., Goodwin, S., Reichenthal, J., Horning, E . , and Brodie, . B . , J .
Pharmacol. Exp. Ther. (1955), 113, 9.
540. Burns, J . J., Yu, T. F., Berger, L . , and Gutmann,
., Amer. J . Med. (1958), 25, 401.
541. Conney, A. H., Trousof, ., and Burns, J . J., J .
Pharmacol. Exp. Ther. (1960), 128, 333.
542. Berger, F. M., J. Pharmacol. Exp. Ther. (1952),
104, 229.
543. Berger, F. M., J . Pharmacol. Exp. Ther. (1952),
104, 468.
544. Editorial, J . Amer. Med. Ass. (1961), 175, 388.
545. Freundt, K. J., Arznein. Forsch. (1973), 23, 949.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

2
Application of the Pro-drug Approach to Antibiotics
A. A. SINKULA
Research Laboratories, The Upjohn Co., Kalamazoo, Mich. 49001

Antibiotics constitute a valuable adjunct to the physicians


therapeutic armamentariu
infectious diseases. Whil
with a minimum of modification to ensure their therapeutic effect,
exceptions exist where extensive development must be undertaken
prior to their becoming efficacious medicinal agents. Certain
shortcomings of these agents, such as lack of stability or poor
bioavailability, can be minimized or eliminated by the use of
carefully designed dosage formulations. In many instances,
however, formulation development fails to improve those properties
of the antibiotic that are necessary to ensure therapeutic efficacy. It is in this area that chemical modification (prodrug
formation) of the parent antibiotic molecule plays an important
role.
In the rational design and synthesis of the ideal antibiotic
prodrug derivative, several factors should be considered and can
be briefly stated as follows:
1. Availability of inexpensive chemical intermediates - any
potential derivative should not substantially enhance production
costs of an already expensive drug. Most starting materials
(acid chlorides or anhydrides, alkyl halides, alkyl or aryl
amines, semicarbazides, etc.) considered as chemical modifiers are
commercially available in a high state of purity at reasonable
costs. Bulk rates are sometimes available on large quantities of
certain starting materials thus further lowering overall costs.
2. Derivative easily synthesized and purified - elaborate
synthetic schemes should be avoided i f at all possible due to
increased costs. Multi-step syntheses increase operator time,
decrease yields of ultimate product, and increase the probability
of unwanted side reactions occurring. Purification should ideally
be effected by crystallization from the reaction mixture. Cumbersome separations such as column or liquid chromatography,
counter-current distribution, etc., should be avoided when
feasible.
116
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

2.

SINKULA

Pro-drug Approach to Antibiotics

117

3. D e r i v a t i v e c o n v e n i e n t l y scaled-up i n high y i e l d - s c a l e up problems i n c r e a s e i n i n t e n s i t y as a f u n c t i o n o f the bench


s c a l e s y n t h e s i s . The s i m p l e r the bench s c a l e scheme, the l e s s
i n v o l v e d are the scale-up problems.
4. D e r i v a t i v e s t a b l e i n bulk form and i n dosage form - many
drug d e r i v a t i v e s l a c k a market due to t h e i r i n s t a b i l i t y i n bulk
or dosage form. Such problems as polymorphic changes, d e g r a d a t i o n
i n the presence o f t r a c e amounts o f moisture o r s o l v a t e , photodecomposition, c a k i n g , melt back, and i n c o m p a t i b i l i t i e s with
v e h i c l e , e x c i p i e n t s , l u b r i c a n t s , e t c . , are common among drug substances. The i d e a l drug d e r i v a t i v e e x h i b i t s s u f f i c i e n t p h y s i c o chemical s t a b i l i t y i n the bulk and formulated s t a t e .
5. D e r i v a t i v e i s s u f f i c i e n t l y l a b i l e i n v i v o - r e g e n e r a t i o n
o f the parent drug molecul i v i v i f
Th m e r i
f
the d e r i v a t i v e p o r t i o n
to modify some u n d e s i r a b l e pharmaceutical (physicochemical)
property o f the parent molecule. I t can a l t e r the t r a n s p o r t ,
d i s t r i b u t i o n , s i t e l o c a l i z a t i o n , metabolism or e x c r e t i o n charact e r i s t i c s o f the parent molecule. Other m o d i f i c a t i o n s can i n c l u d e
increased s o l u b i l i t y ( i n c r e a s e d b i o a v a i l a b i l i t y , decreased pain
on i n j e c t i o n ) , decreased s o l u b i l i t y ( e l i m i n a t i o n o f b i t t e r n e s s or
t a r t n e s s , i n c r e a s e d depot b i o a v a i l a b i l i t y , i n c r e a s e d product
s t a b i l i t y , decreased g a s t r i c or i n t e s t i n a l i r r i t a t i o n ) . For
whatever purpose the drug d e r i v a t i v e i s used, the parent molecule
must be regenerated e i t h e r c h e m i c a l l y (pH e f f e c t s ) and/or enzyma t i c a l l y i n v i v o . In most c a s e s , c h e m i c a l l y b l o c k i n g a f u n c t i o n a l group o f a drug molecule renders the drug t h e r a p e u t i c a l l y
i n a c t i v e , thus, the n e c e s s i t y f o r i n v i v o l a b i l i t y .
Examples o f e n s u r i n g i n v i v o l a b i l i t y o f a drug d e r i v a t i v e
i n c l u d e (a) the use o f " a c t i v a t e d " e s t e r s , e.g. e l e c t r o n w i t h drawing groups adjacent t o the e s t e r bond such as halogens, -NhL*
-N0 ,
R (R=electron withdrawing s u b s t i t u e n t ) and (b)
avoidance o f s t e r i c bulk ( t - b u t y l , i - b u t y l , i - p r o p y l , e t c . ) a t or
near the s i t e o f h y d r o l y s i s .
6. D e r i v a t i v e i s non-toxic - an extremely important cons i d e r a t i o n i n view o f the i n c r e a s e d t o x i c o l o g i c a l t e s t i n g r e q u i r e d f o r any and a l l new promising drug d e r i v a t i v e s . R e l a t i v e l y " s a f e " m o i e t i e s i n c l u d e amino a c i d s , s h o r t t o medium l e n g t h
a l k y l e s t e r s , and many i n o r g a n i c and o r g a n i c a c i d and base s a l t
combinations.
7. The d e r i v a t i v e e x h i b i t s some r e a l advantage over the
parent molecule - prodrug d e r i v a t i v e s , by v i r t u e o f t h e i r a b i l i t y
to c l e a r l y modify some-pharmaceutical p r o p e r t y o f a drug substance, make t h i s a f r u i t f u l area o f drug r e s e a r c h . Advantages
such as i n c r e a s e d a b s o r p t i o n and i n c r e a s e d serum l e v e l s o f parent
drug, l a c k o f pain on i n j e c t i o n , and s u s t a i n e d b i o a c t i v i t y (depot
e f f e c t ) can be claimed. I t should be noted t h a t the m o d i f i c a t i o n
o f one p r o p e r t y f r e q u e n t l y a l t e r s s e v e r a l p r o p e r t i e s o f the drug
molecule and c a u t i o n must be e x e r c i s e d when embarking on a program
of t h i s nature.
2

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

118

PRO-DRUGS

Employing the aforementioned f a c t o r s as a foundation f o r our


r a t i o n a l e , prodrug d e r i v a t i v e s o f s e l e c t e d c l a s s e s o f a n t i b i o t i c s
w i l l be d i s c u s s e d with emphasis on t h e i r chemistry and b i o l o g y .
When p e r t i n e n t , s p e c i f i c examples w i l l be chosen t h a t i l l u s t r a t e
the r a t i o n a l e most e m p h a t i c a l l y .
-Lactam A n t i b i o t i c s
Penicillins.
A m p i c i l l i n . A m p i c i l l i n ( d - a - a m i n o b e n z y l p e n i c i l l i n , V) i s a
broad-spectrum a n t i b i o t i c c u r r e n t l y e n j o y i n g wide use i n a n t i
b a c t e r i a l therapy a g a i n s t a v a r i e t y o f s u s c e p t i b l e g r a m - p o s i t i v e
organisms. While i s r e l a t i v e l s t a b l t stomach pH i t i i n
e f f i c i e n t l y absorbed whe

Chemistry. In an e f f o r t to overcome t h i s a b s o r p t i o n problem,


von Daehne e t . a l . [Vj prepared a s e r i e s o f acyloxymethyl e s t e r s
o f t h i s a n t i b i o t i c . In g e n e r a l , two pathways were u t i l i z e d to
prepare a v a r i e t y o f such e s t e r s (Scheme I ) . The f i r s t route
i n v o l v e s the r e a c t i o n o f potassium b e n z y l p e n i c i l l i n a t e w i t h
chloromethyl p i v a l a t e to form pivaloyloxymethyl b e n z y l p e n i c i l l i
nate 3., f o l l o w e d by h y d r o l y s i s o f the amide s i d e c h a i n with P C I /
q u i n o l i n e to a f f o r d pivaloyloxymethyl 6 - a m i n o p e n i c i l l i n a t e 4.
Treatment o f 4 h y d r o c h l o r i d e with D-a-phenylglycyl c h l o r i d e hydro
c h l o r i d e i n the presence o f sodium bicarbonate g i v e s p i v a l o y l o x y
methyl D - a - a m i n o b e n z y l p e n i c i l l i n a t e 5. ( p i v a m p i c i l l i n ) i n good
y i e l d . 5^ was a l s o prepared from 4_ by u t i l i z i n g a 3-dicarbonyl
p r o t e c t i v e group approach. A mixed anhydride 6_ was prepared by
treatment o f potassium
N-{1-methyl-2-carbethoxyvinyl}-D-a-amino-phenylacetate hemihydrate with i s o b u t y l chloroformate. The
a d d i t i o n o f 4_ to a s o l u t i o n o f the mixed anhydride a f f o r d e d the
a d d i t i o n product ]_ which was hydrolyzed with HC1 i n s i t u to g i v e a
65% y i e l d o f 5.
The t h i r d pathway i n v o l v e s the use o f potassium D-a-azidob e n z y l p e n i c i l l i n a t e 8 with formation o f the e s t e r 9 by treatment
with chloromethyl p i v a l a t e . The a z i d e was c a t a l y t i c a l l y hydrogenerated to _5.
5

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

2.

SINKULA

Pro-drug Approach to Antibiotics

119

B i o l o g i c a l . Although p i v a m p i c i l l i n i s s t a b l e i n n e u t r a l
s o l u t i o n , i t i s r a p i d l y hydrolyzed to a m p i c i l l i n i n the presence
o f e s t e r a s e s d e r i v e d from a v a r i e t y o f mammalian sources ( 2 ) .
Esterases obtained from rodent sources, e.g. r a t and mouse, exh i b i t a high degree of h y d r o l y t i c a c t i v i t y while e s t e r a s e enzymes
o f dog and man show a somewhat lower a c t i v i t y . Table I summarizes
the i n v i t r o enzyme h y d r o l y s i s s t u d i e s conducted with pivampicillin.
These s t u d i e s , while i n d i c a t i v e o f the f a t e o f the e s t e r i n
serum and whole blood, do not provide c o n c l u s i v e proof t h a t the
same e s t e r w i l l behave s i m i l a r l y i n the i n t a c t organism. Human
s u b j e c t s dosed, i n a c r o s s o v e r experiment, with 250 mg. o f amp i c i l l i n and 358 mg. o f p i v a m p i c i l l i n (*250 mg o f a m p i c i l l i n )
showed a b s o r p t i o n o f th
concentrations of ampicilli
The e s t e r was absorbed almost q u a n t i t a t i v e l y (a t h r e e - f o l d i n crease i n peak serum l e v e l s when administered as the e s t e r ) . I t
was f u r t h e r noted t h a t 99% o f the drug i n blood was present as
a m p i c i l l i n 15 minutes a f t e r a d m i n i s t r a t i o n . S p e c u l a t i o n centered
on the f a c t t h a t , due to the inherent l a b i l i t y o f the p i v a l o y loxymethyl e s t e r , h y d r o l y s i s i n v i v o proceeded v i a Sequence I.
A f t e r a b s o r p t i o n o f the e s t e r , h y d r o l y s i s to the hydroxymethyl
e s t e r occurred f o l l o w e d by f u r t h e r degradation to a m p i c i l l i n and
formaldehyde.
Hetacillin. Hetacillin
{6-(2,2-dimethyl-5-oxo-4-phenyl-li m i d a z o l i d i n y l ) p e n i c i l l a n i c acid} V [ represents another type o f
prodrug o f a m p i c i l l i n ( 3 ) .

0
II
T h i s a n t i b i o t i c d e r i v a t i v e was prepared by the condensation o f
acetone with a m p i c i l l i n and was o r i g i n a l l y designed to enhance
the g a s t r o i n t e s t i n a l a b s o r p t i o n of a m p i c i l l i n . I t i s a l s o u t i l i z e d as a s t a b l e form o f a m p i c i l l i n f o r use i n i n f u s i o n s o l u t i o n s
f o r a d m i n i s t r a t i o n over extended periods o f time.
Chemistry. Several methods have been devised whereby H can
be prepared from commercially a v a i l a b l e intermediates (Scheme I I ) .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

103
<1
<1
50
23
50
10
5
<5
5
5
5
3-4

Ha1f-life
(min)

ANTIMICROBIAL AGENTS AND CHEMOTHERAPY


In a l l experiments, the s t a r t i n g c o n c e n t r a t i o n o f p i v a m p i c i l l i n
h y d r o c h l o r i d e was 14.3 yg/ml. Determinations were madeatpH 7.4 and 37 C. (2j

"

None
Mouse serum, 1%
Rat serum, 1%
Dog serum, 5%
Dog serum, 10%
Human serum, 10%
Homogenate o f g a s t r i c mucosa from t h e dog, 10%
Homogenate o f i n t e s t i n a l mucosa from the dog, 10%
L i v e r homogenate from the dog, 10%
Homogenate o f human g a s t r i c mucosa, 10%
Homogenate o f human duodenal mucosa, 10%
Human whole blood
Whole blood from the dog

Enzyme Source

ENZYMATIC HYDROLYSIS OF PIVAMPICILLIN

TABLE I

2.

SINKULA

121

Pro-drug Approach to Antibiotics


1. PCIg/quinoline
2. n-PrOH

HN
2

^3-CH cNH-f-f- 4c:

~^C00CH20C0CMe3

0
0 - CHCCi
H
I
NH2
HCI

O-CH2CNH-1fy:

*0

6.

0E1

0-CHCNH-i-Y ><

C00CH 0C0CMe
2

CICH20C-CMe3
Chloromethylpivolate

Nol
H30+

NH

N_

^C00CH 0C0CMe
2

0
^-CHCNH

<f

0"

-r-f^Y

Scheme I

0
I
/
IIIl

f > C H - c - N :
: .
v

7/

/
CH3
^C00CH 0CC(CH3)
5

Pivampicillin
non-specific esterases
0
Il
n

/rv

2
0

**C00CH 0H + (CH )3C-C00H

<^H-C~N

v i

CH
C

a _ h
3
"
^COOH + C H 0
H

Ampicillin
Antimicrobial Agents and Chemotherapy

Sequence I. Hydrolysis of piOompicillin (2)


In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

122

PRO-DRUGS

The f i r s t r o u t e e n t a i l s the use o f sodium a m p i c i l l i n 10^ and ace


tone, with the formation o f 1J_ being e f f e c t e d under a c i d i c c o n d i t i o n s ( p H 1-3) ( 4 ) . The condensation product p r e c i p i t a t e s from
the aqueous r e a c t i o n mixture and any unreacted a m p i c i l l i n remains
in s o l u t i o n as the HC1 s a l t thereby s i m p l i f y i n g the i s o l a t i o n o f
the d e r i v a t i v e .
A second p r e p a r a t i v e method ( 3 ) i n v o l v e s the r e a c t i o n o f 6a m i n o p e n i c i l l a n i c a c i d J2 w i t h D-(-)--aminophenylacetyl c h l o r i d e
h y d r o c h l o r i d e i n the presence o f acetone a t pH 2.5-3 and low
temperature (0-10). Subsequent pH adjustment t o 7.5 a f t e r
s e v e r a l hours s t a n d i n g , and e x t r a c t i o n with methyl i s o b u t y l
ketone, a f f o r d e d a 50% y i e l d o f U_.
Hetacillin esters I
e f f o r t t f u r t h e enhanc
l
absorption of h e t a c i l l i n
S l e e z e r and Johnson (5)
methoxymethy
lj_ y
the sequence o u t l i n e d i n Scheme I I I . Sodium 6-(a-phenoxyacetamido) p e n i c i l l i n a t e 13 was e s t e r i f i e d a t the C~ carboxyl with
methoxymethyl c h l o r i d e and subsequently d e a c y l a t e d a t C u s i n g
e i t h e r chemical o r enzymatic means t o a f f o r d methoxymetnyl-6aminopen c i l u n a t e JMk The methoxymethyl e s t e r o f a m p i c i l l i n l j [
was produced by r e a c t i n g with D-(-)-a-aminophenylacetyl
c h l o r i d e h y d r o c h l o r i d e . Condensation o f 15 w i t h acetone, under
c o n d i t i o n s d e s c r i b e d p r e v i o u s l y , a f f o r d e d a good y i e l d o f methoxy
methyl h e t a c i l l i n JjS. E s s e r y (6) has s i m i l a r l y prepared the p i v a
loyloxymethyl e s t e r o f h e t a c i l l i n (Scheme II) i n a f u r t h e r e f f o r t
to enhance o r a l a b s o r p t i o n o f t h i s important a n t i b i o t i c .
g

B i o l o g i c a l . Although h e t a c i l l i n i s more s t a b l e than ampi


c i l l i n i n aqueous s o l u t i o n a t a c o n c e n t r a t i o n o f 250 mg./ml.
(<10% d e g r a d a t i o n , 1 hr. vs. 6 h r s . ) , i t appears t h a t h e t a c i l l i n
i s r a p i d l y converted i n v i v o to a m p i c i l l i n ( t ^ 112 minutes) ( 7 ) .
Jusko and Lewis (8) s t u d i e d the pharmacokinetics o f h e t a c i l l i n
and a m p i c i l l i n i n man, and from data generated d u r i n g the study
have d e s c r i b e d the d i s t r i b u t i o n and e l i m i n a t i o n o f t h i s a n t i b i o
t i c u s i n g a two compartment model (Scheme IV). While the model
i s perhaps an o v e r s i m p l i f i c a t i o n , i t can be u t i l i z e d to q u a n t i f y
the d i s t r i b u t i o n and e l i m i n a t i o n parameters o f both a m p i c i l l i n
and h e t a c i l l i n a f t e r intravenous d o s i n g . Table II i l l u s t r a t e s
t h a t f o r those parameters measured, very l i t t l e , i f any, d i f f e r
ence e x i s t s between the magnitude o f the v a l u e s .
The h a l f - l i f e h y d r o l y s i s r a t e f o r h e t a c i l l i n i n v i v o
averages 11.2 minutes (range 8-13 min. f o r 8 s u b j e c t s ) and i t was
s p e c u l a t e d t h a t h y d r o l y s i s might be c h e m i c a l l y r a t h e r than enzyma t i c a l l y mediated. B i o a v a i l a b i l i t y s t u d i e s i n d i c a t e d t h a t the
amount o f the dose absorbed, on the average, was g r e a t e r f o r
h e t a c i l l i n than a m p i c i l l i n (38% vs. 29%). F u r t h e r , a m p i c i l l i n
a b s o r p t i o n was enhanced s l i g h t l y to 42% o f the dose when ad
m i n i s t e r e d as h e t a c i l l i n d u r i n g food i n t a k e but the reason f o r
t h i s i n c r e a s e d a b s o r p t i o n i s not apparent.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

2. SINKULA

-? " !
NH

<f

~~^COONa

CH CCH
3

I. H2O
_ _ _ _ _
3

1 1

r f
H^^CHs ^
N

>

C H

-^0

CICH 0CCMe
2

fV-HC-c'
'
I I

H C CH3

CH3CCH3

^COOH

A_ -cci

N-|-YS<

123

Pro-drug Approach to Antibiotics

r Q
^C00CH 0C0CMe

><

L,

Pivaloyloxymethyl 6 [D(-)-2,2-dimethyl-5-oxo4-phenyl-l-imidazolidinyl] penicillanate

1\
Il
V V0CH C-NH-ir >:

.
Il
(' V o C H ^ - N H - r - r - S Y

P-CcOON.

'

methoxymethyl
chloride

.F^C0OCH 0CH
2

chemical or
enzymatic

H N-f <

S^SST

>*

9>

0
fi^i-CH-CCI
^ N HNH
HCI
2

^-"- TT Y
NH

CH "CH3
3

^C00CH 0CH
^COOCHoOCHx
2

v**

,
*

H C CH
^C00CH 0CH
, , 7/ ~~^Cl
3

Scheme III

HETACILLIN

AMPICILLIN
CENTRAL
COMPARTMENT

|2

*T~~
21

PERIPHERAL
COMPARTMENT

Journal of Pharmaceutical Sciences

Scheme IV. Multiple-compartment phar


macokinetic model used to characterize
hetacillin conversion to ampicillin (k ), am
picillin distribution (k ,k ), and amptcillin
renal (k ) and extrarenal (k ) elimination (8)
c

BILE AND
METABOLISM

URINE

12

21

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

124

PRO-DRUGS

TABLE II
D i s t r i b u t i o n and E l i m i n a t i o n Parameters o f the Two-Compartment
Open Model f o r A m p i c i l l i n
Intravenous
A m p i c i l l i n (SD)

Parameter

A m p i c i l l i n from
H e t a c i l l i n (SD)

D i s t r i b u t i o n volumes, 1
12.0 (1.9)
17.9 (1.5)

12.5 (2.8)
19.3 (2.9)

341 (91)
335 (56)

296 (91)
350 (102)

0.384 (0.185)
0.733 (0.163)
1.73 (0.49)
1.55 (0.47)
0.17 (0.12)
1.29 (0.11)

0.419 (0.180)
0.728 (0.161)
1.68 (0.30)
1.58 (0.31)
0.10 (0.13)
1.34 (0.20)

0.899 (0.075)

0.939 (0.076)

22.7 (5.2)

22.9 (7.6)

0.581 (0.133)
0.297 (0.081)
0.878 (0.151)

0.581 (0.118)
0.355 (0.102)
0.921 (0.141)

Clearances, ml./mi.
Rate c o n s t a n t s , hr.
k
k

1 2
2 1

pi
e
b
Slow th hr.
Fraction (f )
excreted i n u r i n e
Plasma l e v e l
_-.
a r e a , meg. hr. ml.
Integral c o e f f i c i e n t s , hr.
k
K

JOURNAL OF PHARMACEUTICAL SCIENCES


V , VpSS, C l g , C 1 , and area are normalized f o r 1.73 m. body
s u r f a c e area. (8).
a

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

2.

SINKULA

125

Pro-drug Approach to Antibiotics

C a r b e n i c i l l i n . C a r b e n i c i l l i n (-carboxybenzylpenicillin)
17
i s an important p e n i c i l l i n analog having unique b i o a c t i v i t y
a g a i n s t pseudomonas aeruginosa and i n d o l e - p o s i t i v e Proteus species
which are u s u a l l y r e s i s t a n t to a m p i c i l l i n . C a r b e n i c i l l i n i s
u s u a l l y administered p a r e n t e r a l l y due to i t s poor g a s t r o i n t e s t i n a l
absorption c h a r a c t e r i s t i c s . A d d i t i o n a l l y , i t i s rendered i n a c
t i v e i n the g a s t r i c contents due to i t s a c i d l a b i l i t y . Many
d e r i v a t i v e s , p r i m a r i l y e s t e r s , of t h i s , a n t i b i o t i c have been pre
pared i n an attempt to overcome these shortcomings inherent i n
the parent molecule.

The e s t e r most widely studied to date i s c a r b e n i c i l l i n i n danyl sodium - sodium 6-{2-phenyl-2-(5-indanyloxycarbonyl)}


acetamido p e n i c i l l i n a t e 19.
Chemistry. The s y n t h e s i s of 19 i s o u t l i n e d i n Scheme V and
was devised by Hobbs (_) as a r a d i o s y n t h e s i s u t i l i z i n g H l a b e l e d
i n d a n o l . The mnoacid c h l o r i d e of phenylmalonic a c i d was prepared
by treatment of phenylmalonic a c i d with t h i o n y l c h l o r i d e i n r e f l u x i n g dimethylformamide. The a c i d c h l o r i d e was condensed with
indanol to y i e l d the indanyl e s t e r of phenylmalonic a c i d 18 i n 70%
y i e l d . The a c i d c h l o r i d e o f |8 was prepared by a d d i t i o n of
t h i o n y l c h l o r i d e and was subsequently t r e a t e d with 6-aminopenic i l l a n i c a c i d to a f f o r d 1J.
A s e r i e s o f 3-acyloxyalkyl e s t e r s was synthesized by B u t l e r
and Hamanaka (10) as l a b i l e , l i p o p h i l i c d e r i v a t i v e s of c a r b e n i c i l l i n designed to enhance the o r a l absorption o f c a r b e n i c i l l i n .
To produce the 3-mono(a-acetoxyethyl) e s t e r 2]_, a-chloroethyl
acetate i s added to a suspension o f 6 - a m i n o p e n i c i l l a n i c a c i d ]2_
(as the t r i e t h y l a m i n e s a l t ) and s t i r r e d f o r several hours.
(Scheme VI) T h i s e s t e r 20 can be i s o l a t e d and stored f o r f u t u r e
use as the p - t o l u e n e s u l f o n i c a c i d s a l t or i t can be converted
d i r e c t l y to 21_ by a c y l a t i o n with phenylmalonic a c i d mono a c i d
c h l o r i d e . T h i s l a s t step i n the r e a c t i o n sequence proceeds
o p t i m a l l y i n a heterogeneous s o l v e n t system employing water and a
water immiscible i n e r t s o l v e n t such as i s o p r o p y l ether or benzene
at i c e - b a t h temperature. As the product forms, i t p a r t i t i o n s
i n t o the organic l a y e r and can be d r i e d and recovered i n high
yields.
3

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

126

PRO-DRUGS

a-{Carbo(a-acetoxyethyloxy)} b e n z y l p e n i c i l l a n i c a c i d 21a can


be prepared d i r e c t l y by a c y l a t i o n of 12 with a-acetoxyethyl
phenylmalonyl c h l o r i d e 22 (11). (Scheme V I I ) . T h i s r e a c t i o n step
i s f a c i l i t a t e d by employing a 20-40% molar excess o f a c i d c h l o r i d e
i n a heterogeneous s o l v e n t system at a pH of 5.5 - 6.5.
B i s e s t e r s o f c a r b e n i c i l l i n 24 have i d e a l l y been made by
d i r e c t e s t e r i f i c a t i o n o f c a r b e n i c i l l i n disodium s a l t 23. with the
d e s i r e d -acyloxyalkyl h a l i d e (Scheme V I I I ) .
B i o l o g i c a l . In v i t r o - the minimal i n h i b i t o r y c o n c e n t r a t i o n
(MIC) o f c a r b e n i c i l l i n indanyl sodium was determined by a s e r i a l
d i l u t i o n technique using a v a r i e t y o f b a c t e r i a l i s o l a t e s o f
c l i n i c a l o r i g i n (12). The MIC was found to be s i m i l a r to c a r
b e n i c i l l i n but may be misleading s i n c e the c o n d i t i o n s o f the
assay ( i n c u b a t i o n at 3
could cause h y d r o l y s i s o
S t a b i l i t y i n a c i d i c media. A c i d s t a b i l i t y i s r e p u t e d l y one
of the major advantages o f c a r b e n i c i l l i n e s t e r s . Studies de
signed to t e s t t h i s premise were performed by i n c u b a t i n g the i n
danyl e s t e r i n s y n t h e t i c g a s t r i c j u i c e (pH2) at 37 C. f o r one
hour. There was no l o s s o f a c t i v i t y ( r e f l e c t i n g a c i d s t a b i l i t y )
f o r c a r b e n i c i l l i n indanyl sodium ]_9 while disodium c a r b e n i c i l l i n
23 l o s t 99.2% of i t s a n t i b a c t e r i a l a c t i v i t y .
C a r b e n i c i l l i n indanyl e s t e r i s the d e r i v a t i v e t h a t has
been s t u d i e d most i n t e n s i v e l y i n several mammalian species (9).
In r a t s , absorption of 19 i s v i r t u a l l y q u a n t i t a t i v e . Using r a d i o
l a b e l e d indanyl c a r b e n i c i l l i n , >99% of the dose i s excreted v i a
the u r i n e i n 24 hours. Only t r a c e s o f r a d i o a c t i v i t y are found i n
the feces i n d i c a t i n g t h a t t h i s e s t e r d e r i v a t i v e g r e a t l y enhances
a b s o r p t i o n o f c a r b e n i c i l l i n . The e s t e r i s r a p i d l y hydrolyzed
a f t e r absorption and the l a b e l e d indanol i s excreted as the
glucuronide and s u l f a t e conjugates. Using the dog as the b i o
l o g i c a l model f o r absorption s t u d i e s , the absorption and excre
t i o n patterns are more complex. Assay o f dog u r i n e a f t e r admini
s t r a t i o n of indanyl c a r b e n i c i l l i n i n d i c a t e that about 20% o f the
dose presented as indanol conjugates. The remainder appear as
other conjugated metabolites o f indanol (Scheme IX) i n the form
o f hydroxy indanols and hydroxy indanones.
B i o a v a i l a b i l i t y s t u d i e s c a r r i e d out with 12 human v o l u n t e e r s ,
who were administered a s i n g l e one gram dose, p a r a l l e l e d the ab
s o r p t i o n and e x c r e t i o n p a t t e r n found i n r a t s (9). Thus, t h a t
amount of indanol (as the glucuronide and s u l f a t e conjugates)
t h e o r e t i c a l l y a t t r i b u t a b l e to the amount o f e s t e r administered
was accounted f o r i n the u r i n e . The indanyl e s t e r appears to be
r a p i d l y absorbed q u a n t i t a t i v e l y from the g a s t r o i n t e s t i n a l t r a c t
with subsequent h y d r o l y s i s by n o n - s p e c i f i c serum and t i s s u e
e s t e r a s e s to c a r b e n i c i l l i n .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

SINKULA

127

Pro-drug Approach to Antibiotics

J!
Scheme V

CO H
Z

f 3
---

CH C00CHCI

\oC\

HaN-r-fSC

J-N-<C02H
0

-CH

TEA

J-N^ 00CH0C0CH
0
j
C

12.

20

< ^ C H - C N H y - f
*

C0 H

CH3

~~^C00CH0C0CH
CI H
3

21

Scheme VI

-CHCOCI
. =0
^
CH0C0CH
I
CH

^ C H - C - N H ^ S ^

M ~~^C00H

A= 0

CH0C0CH
I
CH

22

21

Scheme VII

CH3

<_ ~"~ _
C02Na

/ ~~^C02Na
0

2CH3C00CHC1

0- -"- t_

23

nh

M
0

C=0
,
0
f
CH

"C0 CH0C0CH
,
2

CH
C

24

Scheme VIII
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

128

PRO-DRUGS

S i m i l a r claims have been made f o r a v a r i e t y o f s u b s t i t u t e d


t h i e n y l e s t e r s o f c a r b e n i c i l l i n (13).
Other prodrug d e r i v a t i v e s o f a v a r i e t y o f p e n i c i l l i n s have
been prepared u t i l i z i n g b a s i c a l l y t h e chemical pathways and
r a t i o n a l e d i s c u s s e d above and a r e summarized i n Table I I I .
Cephalosporins. The c e p h a l o s p o r i n s , a new c l a s s o f a n t i
b i o t i c s c h e m i c a l l y s i m i l a r t o t h e p e n i c i l l i n s , possess advantages
not i n h e r e n t i n many o f the p e n i c i l l i n d e r i v a t i v e s . Although
both c l a s s e s o f a n t i b i o t i c s share a commonality i n the presence
o f a 3-lactam r i n g , the c e p h a l o s p o r i n s c o n t a i n a six-membered
d i h y d r o t h i a z i n e r i n g i n l i e u o f the five-membered t h i a z o l i d i n e
r i n g present i n t h e p e n i c i l l i n s . The c e p h a l o s p o r i n s e x h i b i t a
high degree o f r e s i s t a n c e t o p e n i c i l l i n a s e - p r o d u c i n g s t a p h l o
c o c c i (35), possess b a c t e r i c i d a
and g r a m - p o s i t i v e b a c t e r i (36)
cross-allergenicity
with p e n i c i l l i n (37). Several c e p h a l o s p o r i n s c u r r e n t l y e n j o y i n g
c l i n i c a l acceptance i n c l u d e c e p h a l o t h i n , c e p h a l o r i d i n e , cephalog l y c i n and c e p h a l e x i n .
Despite the f a c t t h a t t h e c e p h a l o s p o r i n s e x h i b i t adequate
s t a b i l i t y i n a c i d i c media, they a r e p o o r l y absorbed on o r a l ad
m i n i s t r a t i o n . C e p h a l o g l y c i n and c e p h a l e x i n , however, have
demonstrated higher serum l e v e l s than c e p h a l o t h i n and c e p h a l o r i
d i n e (38-41). Various attempts have been made t o enhance the o r a l
a b s o r p t i o n o f t h e c l i n i c a l l y u s e f u l c e p h a l o s p o r i n s by r e v e r s i b l e
m o d i f i c a t i o n s ( u s u a l l y e s t e r i f i c a t i o n ) a t the C3 and C4 p o s i t i o n s
on t h e d i h y d r o t h i a z i n e r i n g . The s y n t h e t i c approaches u t i l i z e d
to o b t a i n l a b i l e d e r i v a t i v e s o f these a n t i b i o t i c s i s both imagina
t i v e and voluminous and an e f f o r t w i l l be made here o n l y t o high
l i g h t some o f the more s u c c e s s f u l accomplishments.
Specific
examples w i l l be used throughout the d i s c u s s i o n .
Chemistry - C3 e s t e r s ( c e p h a l o t h i n ) . The achievements o f
Flynn (42) and Kukolja (43,44) exemplify t h e unique s y n t h e t i c
routes taken t o o b t a i n C3 e s t e r s . The i n i t i a l step i n v o l v e s
the p r e p a r a t i o n o f 7-(2'-thienylacetamido)-3-hydroxymethyl-A2cephem-4-carboxylic a c i d 26 by the simultaneous h y d r o l y s i s and
isomerization o f 7-(2 -thienylacetamido) cephalosporanic acid
( c e p h a l o t h i n , 25_) with sodium hydroxide. (Scheme X). 26 was
then e s t e r i f i e d with cyclobutane c a r b o x y l i c anhydride i n p y r i d i n e
to y i e l d
7-(2'thienylacetamido)-3-cyclobutylcarbonyloxymethyl- cephem-4-carboxylic a c i d 27. I s o m e r i z a t i o n t o the e s t e r was
accomplished by warming equimolar q u a n t i t i e s o f 27 and mc h l o r o p e r b e n z o i c a c i d f o r 10 minutes. I s o l a t i o n o f 7 - ( 2 ' - t h i e n y l acetamido)-3-cyclobutylcarbonyloxymethyl-A -cephem-l-oxide-4c a r b o x y l i c a c i d 28 as c r y s t a l s was achieved by evaporating t h e
m a j o r i t y o f t h e s o l v e n t . Reduction o f t h e 1-oxide with stannous
c h l o r i d e and a c e t y l c h l o r i d e y i e l d e d 7-(2'thienylacetamido)-3cyclobutylcarbonyloxymethyl-A -cephem-4-carboxylic a c i d 29.
,

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

SINKULA

Pro-drug Approach to Antibiotics


20%
HO"

OH
15%
HO

<0

HO

^0

I
OH

iJO
Antimicrobial Agents and

10%

Chemotherapy

Scheme IX. Indanol metabolites in dog urine


after administration of indanol or indanyl carbenicillin. The figures represent the rehtive
amounts of each substance present. All are
present in urine as their glucuronide and sulfate
ester conjugates (9).

Scheme X
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

7. 6-(D--Su1foami nopheny 1acetamido) p e n i c i l l i n


Pivaloyloxymethyl e s t e r

Oral

25
Absorption

26

17,18
1,2,19-22
3,8,23
32
24

Absorption
Absorption
Absorption
Absorption
Absorption

Absorption

16

15

Absorption

Oral
Absorption

Reference
14

Property
Modified
Absorption

Route o f
Administration
Oral

3. P e n i c i l l i n , general s t r u c t u r e D i e t h y l aminoethyl e s t e r s ,
Oral
alkoxymethyl e s t e r s , ether
O r a l , IV
a. Azide
4. -Aminobenzyl p e n i c i l l i n
b. Acyloxymethyl e s t e r s
Oral
c.
N,N-isopropylidene
Oral
adduct
Oral
d. P h t h a l i d y l e s t e r
IM
Amide
5. P e n i c i l l i n G
Penicillin V
6. -Amino ( o r ureido)
Acyloxymethyl e s t e r s
Oral
cyclohexad i e n y l a l k y l
penicillin

esters

Chemical
Modification
Pivaloyloxymethyl e s t e r

2. -Aryl-3-aminoethyl peni c i 11i Alkoxymethyl

Parent Molecule
1. 6-N '-Cyanoamidopeni c i 11i

TABLE I I I . PENICILLIN PRODRUGS DESIGNED TO MODIFY VARIOUS PROPERTIES


OF THE PARENT ANTIBIOTIC

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

Carbenicillin

9.

Pivaloyloxymethyl-D-aami nobenzylpeni c i 11i nate

14. P e n i c i l l i n , general s t r u c t u r e

13.

12. -Aminobenzyl p e n i c i l l i n

11. A c e t a m i d o p e n i c i l l i n s

penicillin

10. 6-(3-Thienyloxyacetamido)

Hetacillin

Molecule

8.

Parent

Chalcon-4-yl
Amide

b.

salt

a.

Probenecid

Dibenzyl ethylene
diamine s a l t

Carboxamido e s t e r

esters

Resistance t o p e n i
cillinase
Resistance to peni
cillinase

Oral
Oral

Bitterness

Duration o f a c t i v i t y

Duration o f a c t i v i t y

Oral

IM

IM

Oral

34

33

30

29

28

31

10
9,12
27

Absorption
Absorption
Absorption
Absorption,
duration of a c t i v i t y

Absorption

Oral

Reference

Property
Modified

Route o f
Administration

Mono and b i s a l k y l e s t e r s Oral


Oral
Indanyl e s t e r
Oral
Thienyl esters

ester

Acetoxymethyl e s t e r

a.
b.
c.

Pivaloyloxymethyl

Chemical
Modification

TABLE I I I . (Continued)

00

ST
*

QTQ

>

132

PRO-DRUGS

An a l t e r n a t i v e procedure (Scheme XI) i n v o l v e s d e a c e t y l a t i o n o f the


potassium s a l t o f 25 with c i t r u s a c e t y l e s t e r a s e (orange peel
enzyme) to a f f o r d potassium 7-(2'-thienylacetamido) cepbalosporadesate* 30 i n good y i e l d without i s o m e r i z a t i o n to the
d e r i v a t i v e (45,46). A c y l a t i o n o f the 3-hydroxymethyl group i s
base dependent. Conventional attempts a t
a c y l a t i o n under a c i d i c c o n d i t i o n s , e.g. a c e t i c anhydride, pro
duce cephalosporadesolactones. Aromatic e s t e r d e r i v a t i v e s are
made, however, by employing the c o n d i t i o n s o f the SchottenBaumann r e a c t i o n . Thus, treatment o f 30 with a l a r g e excess
of benzoyl c h l o r i d e and sodium hydroxide i n aqueous acetone
a f f o r d s a good y i e l d o f sodium 0-benzoyl-7-(2-thienylacetamido)
cephalosporadesate 3]_. A l i p h a t i c a c i d c h l o r i d e s under the same
c o n d i t i o n s r e a c t p r e f e r e n t i a l l with
d
esterificatio
occurs.
C4 e s t e r s . The c e p h a l o s p o r i n C4 e s t e r s are s y n t h e s i z e d
by conventional methods (47). (Scheme X I I ) . A d d i t i o n o f e q u i molar q u a n t i t i e s o f trietTiylamine (TEA) and i s o b u t y l c h l o r o f o r mate to 25 gave 7 - ( 2 - t h i e n y l a c e t a m i d o ) c e p h a l o s p o r a n i c a c i d monoi s o b u t y l c a r b o n a t e anhydride 32 as an o i l which was subsequently
obtained c r y s t a l l i n e . The e t h y l carbonate anhydride was a l s o
prepared by t h i s method. F u r t h e r attempts t o e s t e r i f y a t C4 v i a
the mixed anhydride r e s u l t e d i n mixtures o f and e s t e r s .
E f f o r t s t o separate the i s o m e r i c e s t e r s by r e c r y s t a l l i z a t i o n
were u n s u c c e s s f u l .
f

Ca amides. Two approaches have been u t i l i z e d i n an e f f o r t


to form c e p h a l o s p o r i n C4 amides. The f i r s t i n v o l v e s the d i r e c t
condensation o f c e p h a l o t h i n with
,'-dicyclohexylcarbodiimide
(DCC) to form the a c t i v a t e d carboxyl i n t e r m e d i a t e . (Scheme X I I I ) .
The exchange r e a c t i o n with t-butyl--aminopropionate t o form
N - { l - c a r b o - t - b u t o x y ) e t h y l } - 7 - ( 2 ' - t h i e n y l a c e t a m i d o ) cephalospor
a n i c a c i d amide 33 proceeds without i s o m e r i z a t i o n . The a l t e r n a
t i v e approach i s somewhat more i n v o l v e d and p r o v i d e s the C4
amide as the isomer. (Scheme XIV) (47). The procedure
of Nefkens e t . a l . (48) was f o l l o w e d t o produce the a c t i v a t e d c a r bonyl i n t e r m e d i a t e . Thus, 25, N-hydroxyphthalimide (phthaloxime,
34) and DCC were s t i r r e d t o g e t h e r and s t o r e d f o r s e v e r a l days.
Work up o f the r e a c t i o n mixture gave a 50% y i e l d o f N-{7-(2't h i e n y l a c e t a m i d o ) cephalosporanoyloxy} phthalimide 35. The r e a c
t i o n o f 35 with e t h y l g l y c i n a t e a f f o r d e d a 73% y i e l d o f completely
isomerized tr amide, N-(carbethoxymethyl)-3-acetoxymethyl-7-(2*thienylacetamido)-2-cephem-4-carboxylic a c i d amide 3j6. Other
2

D e s a c e t y l c e p h a l o s p o r i n s have been t r i v i a l l y named cephalosp o r a d e s i c a c i d s f o r convenience.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

SINKULA

25

Pro-drug Approach to Antibiotics

^CHCH 0C0CI

-I^U

CH2CNH
i-Nvj^CH 0C0CH .
C00C00CH Crf
^CH

3 ct

32

Scheme XII

1. DCC
2. CH CHC00C(CH
,
" ' )3
'
VV

N H

C H

C N H

T /-N>f^CH20C0CH3
T J
0

C0NHCHC00C(CH3)3
33

CH3

Scheme XIII

8
JVW^^

CH CNH-p^
2

c H 2 0 c o c H 3

C0NHCH C00C H
2

36

Scheme XIV
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

134

PRO-DRUGS

amino a c i d e s t e r s r e a c t i n s i m i l a r f a s h i o n .
Another s e r i e s o f i n t e r e s t i n g s y n t h e t i c r o u t e s designed t o
p r o v i d e b i o r e v e r s i b l e Ca c e p h a l o s p o r i n e s t e r s a r e r e p o r t e d by
Binderup e t . a l . (49). S t i m u l a t e d by the success achieved with
acyloxymethyl e s t e r s o f a m p i c i l l i n , these i n v e s t i g a t o r s attempted
to repeat the e a r l i e r achievements with c e p h a l o g l y c i n . U t i l i z i n g
the sodium s a l t o f c e p h a l o t h i n as t h e i r s t a r t i n g p o i n t , t h e
and acetoxymethyl e s t e r s were made by treatment with c h l o r o
methyl a c e t a t e . (Scheme XV). T h i s mixture was o x i d i z e d t o the
s u l f o x i d e 37, as p r e v i o u s l y d e s c r i b e d , and subsequently reduced
to acetoxymethyl-7-(2'-thienylacetamido) cephalosporanate 38 with
phosphorus t r i c h l o r i d e .
Acetoxymethyl-7-aminocephalosporanate
39 was produced by treatment o f 38 with phosphorus p e n t a c h l o r i d e
and n-propanol. The h y d r o c h l o r i d e s a l t o f 39 was formed by the
a d d i t i o n o f IN HC1. T
chloride to yield
acetoxymethyl-7-(D-a
cephalosporanate 40 which was subsequently hydrogenated with 10%
palladium/carbon t o y i e l d
acetoxymethyl-7-(D-a-aminophenylacetamido) cephalosporanate 4K
The p i v a l o y l o x y m e t h y l e s t e r was s y n t h e s i z e d by combining
f e a t u r e s o f s e v e r a l syntheses p r e v i o u s l y d e s c r i b e d . T h i s e s t e r
was i n i t i a l l y prepared by treatment o f potassium 7-(D-a-azidophenylacetamido) cephalosporanate 42 with c h l o r o m e t h y l p i v a l a t e .
A mixture o f t h e 43 and e s t e r s were formed by t h i s proce
dure. Treatment o f the mixture f i r s t with m-chloroperbenzoic
a c i d t o form t h e s u l f o x i d e e s t e r and secondly with sodium
d i t h i o n i t e / a c e t y l c h l o r i d e gave t h e r e q u i s i t e p i v a l o y l o x y m e t h y l 7-(D-a-azidophenylacetamido) cephalosporanate 44 e x c l u s i v e l y as
the isomer. Reduction o f 44 by c a t a l y t i c hydrognation a f forded pivaloyloxymethyl-7-(D-a-aminophenylacetamido)
cephalosporanate 45.
Many o t h e r c e p h a l o s p o r i n d e r i v a t i v e s have been made u t i l i z i n g s y n t h e t i c pathways s i m i l a r t o those p r e v i o u s l y d i s c u s s e d
(50-53).
2

B i o l o g i c a l . Several attempts have been made t o improve


c e r t a i n physicochemical and b i o l o g i c a l p r o p e r t i e s ( e s p e c i a l l y
61 a b s o r p t i o n ) o f t h e c e p h a l o s p o r i n s by the prodrug approach.
Most e f f o r t s have been o n l y moderately s u c c e s s f u l . Kukolja (44)
r e p l a c e d the 3-acetate o f c e p h a l o t h i n with a s e r i e s o f s t e r i c a l l y
hindered e s t e r s f o r t h e purpose o f i n h i b i t i n g h y d r o l y s i s a t t h i s
p o s i t i o n on the a n t i b i o t i c . The b u t y r a t e and i s o b u t y r a t e d e r i v a t i v e s e x h i b i t e d good i n v i t r o a c t i v i t y a g a i n s t a v a r i e t y o f gramp o s i t i v e and gram-negative b a c t e r i a . ED50 values i n mice i n d i c a t e d b i o a c t i v i t y somewhat improved over sodium c e p h a l o t h i n .
The c y c l o b u t y r a t e d e r i v a t i v e a l s o e x h i b i t e d good broad-spectrum
b i o a c t i v i t y . Chauvette and Flynn (47) s y n t h e s i z e d a v a r i e t y o f
C4 e s t e r s and amides o f c e p h a l o t h i n with the s p e c i f i c o b j e c t i v e
of o b t a i n i n g d e r i v a t i v e s with improved o r a l a b s o r p t i o n . On adm i n i s t r a t i o n o f these d e r i v a t i v e s t o mice, a low order o f a n t i -

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

2.

SINKULA

Pro-drug Approach to Antibiotics

135

b a c t e r i a l a c t i v i t y was found (as measured by l e v e l s o f a n t i b i o t i c


i n the blood).
A s e r i e s of C3 a r o y l d e r i v a t i v e s o f c e p h a l o t h i n and 7-phenylmercaptoacetamidocephalosporanic a c i d were made by Van Heyningen
(46) and found to possess no s i g n i f i c a n t l y improved b i o a c t i v i t y
over the parent a n t i b i o t i c s .
The s y n t h e t i c and b i o a c t i v i t y s t u d i e s o f Binderup e t . a l . (49)
with the acyloxymethyl e s t e r s of c e p h a l o g l y c i n represent the
f i r s t major success i n o b t a i n i n g a s u p e r i o r o r a l l y absorbed prodrug o f a cephalosporin. The s y n t h e s i s of the C4 acetoxymethyl
e s t e r 41_ and the pivaloyloxymethyl e s t e r o f c e p h a l o g l y c i n 45 are
d e t a i l e d i n Scheme XVI. The h a l f - l i v e s of 4]_ and 45 i n 10% human
serum are 5 minutes and 10-20 minutes r e s p e c t i v e l y . The absorpt i o n and e x c r e t i o n pattern
over study i n f o u r f a s t i n g
e f f i c i e n t absorption and r a p i d e s t e r h y d r o l y s i s a f t e r a b s o r p t i o n .
F i g u r e 1 i l l u s t r a t e s serum l e v e l s obtained with these e s t e r s
vs. c e p h a l o g l y c i n . Recovery of c e p h a l o g l y c i n i n the u r i n e during
s i x hours a f t e r a d m i n i s t r a t i o n represented 68% (acetoxymethyl
e s t e r ) and 61% (pivaloyloxymethyl e s t e r ) o f the t h e o r e t i c a l
amount administered. The corresponding average f i g u r e f o r cepha l o g l y c i n was 18%.
Cephalosporins viewed as prodrugs. When administered to
animals and man, s e v e r a l of the cephalosporins are metabolized
to the correspondingly b i o a c t i v e d e s a c e t y l c e p h a l o s p o r i n .
These C3 acyloxymethyl e s t e r s can, t h e r e f o r e , be considered
cephalosporin prodrugs, e.g., c e p h a l o t h i n i s e n z y m a t i c a l l y and/or
c h e m i c a l l y hydrolyzed i n v i v o to the b i o a c t i v e d e s a c e t y l c e p h a l o t h i n (54,55). F u r t h e r , the lactones of c e r t a i n d e s a c e t y l cephalos p o r i n s e x h i b i t a c t i v i t y a g a i n s t a s t r a i n of Staphylococcus
aureus t h a t i s equal to t h a t o f the parent cephalosporin (56),
i n d i c a t i n g t h a t perhaps the lactone may a l s o be considered a
cephalosporin prodrug.
Studies on the metabolic f a t e o f c e p h a l o g l y c i n i n the r a t by
S u l l i v a n and coworkers (57) have demonstrated t h a t a l a r g e
amount o f the o r a l absorbed dose o f t h i s a n t i b i o t i c i s excreted
as d e s a c e t y l c e p h a l o g l y c i n . Approximately 70% of the administered
dose i s recovered i n the feces i n d i c a t i n g poor GI a b s o r p t i o n .
P a r e n t e r a l l y administered c e p h a l o g l y c i n i s a l s o metabolized
p r i m a r i l y to d e s a c e t y l c e p h a l o g l y c i n . In the mouse, Wick, e t . a l .
(58) found o r a l l y administered c e p h a l o g l y c i n i n u r i n e i n a 1:1
r a t i o with d e s a c e t y l c e p h a l o g l y c i n . The p r i n c i p a l metabolite o f
c e p h a l o g l y c i n i n humans i s d e s a c e t y l c e p h a l o g l y c i n and i s equival e n t i n a c t i v i t y a g a i n s t gram-positive organisms but i s l e s s
a c t i v e a g a i n s t gram-negative organisms. E r a d i c a t i o n of u r i n a r y
t r a c t i n f e c t i o n s i s a t t r i b u t e d mainly to the b i o a c t i v i t y o f
the d e s a c e t y l d e r i v a t i v e suggesting t h a t c e p h a l o g l y c i n may be the
prodrug d e r i v a t i v e of d e s a c e t y l c e p h a l o g l y c i n .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

136

PRO-DRUGS

CO3H

0
O Ss . CH CNH-|f |
/-Nxj^
0
frtrtUn
b N

CICH20C0CH3
Chloromethylacetate

C H 2 0 A c

5 ,

0
f
^

ff\
^ c A ,

PCI3

* N-f^CH 0Ac
C00CH 0C0CH3
L

J~ Y^CH 0Ac
COCH2OCOCH3
II
0
N


0
VAru-Lu

0
J

L PCI /quinoline
2. PrOH
5

^S

/)-N^CH 0Ac
C00CH 0C0CH3
2

S^CH CNH-r-Y h

3. H 0
3

37

- f "h
H 2 N- - r!
// N>f^CH 0Ac

-HC0C

U2

d'C00CH 0C0CH3
2

11 _

J-Nf^CH 0Ac
2

39

fnnrMonmnHi

0
NH

/r Y^CH 0Ac
C00CH 0C0CH3
N

2
0

il
Scheme XV

Scheme XVI
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

2.

SINKULA

Pro-drug Approach to Antibiotics

137

Table IV contains s e v e r a l a d d i t i o n a l cephalosporin prodrugs


designed f o r a v a r i e t y o f uses i n medical p r a c t i c e .
Rifampicin
The r i f a m y c i n s , a f a m i l y of a n t i b i o t i c s i s o l a t e d from the
fermentation broth o f Streptomyces mediterranei n. sp., are c u r
r e n t l y used i n c l i n i c a l p r a c t i c e a g a i n s t gram-positive micro
organisms and t u b e r c u l a r i n f e c t i o n s (mycobacteria). Of s e v e r a l
r i f a m y c i n s , r i f a m y c i n SV and r i f a m y c i n d i e t h y l a m i d e are used
p a r e n t e r a l l y a g a i n s t these b a c t e r i a l i n f e c t i o n s . Rifamycin SV,
moreover, achieves extremely high b i l e c o n c e n t r a t i o n s and i s
used i n i n f e c t i o n s o f the b i l i a r y t r a c t . The a v a i l a b i l i t y o f
3-formyl r i f a m y c i n SV 46 l e d to the s y n t h e s i s o f a l a r g e number
of rifamycin d e r i v a t i v e
form of t h i s a n t i b i o t i
{3-(4-methyl
p i p e r a z i n y l i m i n o m e t h y l ) r i f a m y c i n SV}, 47, ( r i f a m p i c i n ) p r o t e c t s
mice a g a i n s t experimental s t a p h y l o c o c c a l i n f e c t i o n s a t low o r a l
doses (=0.1 mg./kg.). In man, i t i s o f low t o x i c i t y and well
absorbed o r a l l y (73-75). While e l i m i n a t i o n occurs mainly
through the b i l e T e n t e r o h e p a t i c c i r c u l a t i o n ) , small amounts are
a l s o found i n u r i n e . Thin l a y e r chromatography s t u d i e s i n d i c a t e d
t h a t 47 was almost e n t i r e l y converted to d e s a c e t y l r i f a m p i c i n 48
i n v i v o and was probably the a c t i v e a n t i b a c t e r i a l form o f t h i s
a n t i b i o t i c . Thus, while i r r e v e r s i b l e m o d i f i c a t i o n o f 3-formyl
r i f a m y c i n SV to r i f a m p i c i n enhances o r a l a b s o r p t i o n , i n v i v o
d e a c e t y l a t i o n o f the C25 a c e t a t e l i b e r a t e s 48, the t r u e parent
b i o a c t i v e s p e c i e s o f t h i s a n t i b i o t i c . (Scheme XVII). R i f a m p i c i n ,
then, represents the prodrug form o f d e s a c e t y l r i f a m p i c i n . T h i s
f a c t i s c o r r o b o r a t e d by a n t i b a c t e r i a l s t u d i e s with d e s a c e t y l r i f a m p i c i n i n man i n which the b i o a c t i v i t y i s found to be
e x c e l l e n t a g a i n s t gram-positive b a c t e r i a (76). Oral a d m i n i s t r a
t i o n o f a 150 mg. dose o f r i f a m p i c i n i n man produces b i l e l e v e l s
o f d e s a c e t y l r i f a m p i c i n i n excess o f 95% over a 5 hour p e r i o d .
No other m e t a b o l i t e s are present (77).
Clindamycin
Clindamycin {7(S)-chloro-7-deoxylincomycin} h y d r o c h l o r i d e
49 i s a s e m i s y n t h e t i c a n t i b i o t i c d e r i v e d from l i n c o m y c i n . I t s
a c t i v i t y a g a i n s t gram-positive aerobes and gram-positive and
gram-negative anaerobic pathogens i s g r e a t e r than t h a t o f
l i n c o m y c i n . Clindamycin i s well absorbed from the GI t r a c t and
produces serum l e v e l s c o n s i d e r a b l y g r e a t e r than lincomycin (7880). The extreme b i t t e r n e s s o f clindamycin precludes i t s s o l u
t i o n or suspension f o r m u l a t i o n as an acceptable o r a l dosage form.
F u r t h e r , the i n c i d e n c e o f pain a t the i n j e c t i o n s i t e a f t e r i n t r a
muscular i n j e c t i o n i s c o n s i d e r a b l e . These u n d e s i r a b l e p r o p e r t i e s
o f clindamycin prompted a search f o r prodrug d e r i v a t i v e s o f
t h i s a n t i b i o t i c designed t o enhance i t s a c c e p t a b i l i t y .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

4. 7-Acyl ami nodesacetoxy


cephalosporanic a c i d
5. 7-Acylcephalosporani c
acid
C^-benzhydryl

esters

esters

C^-acyloxybenzyl

Oral

Oral

62
63

Absorption
Absorption

61

Absorption,
decreased
toxicity
Oral

C^-acyloxyalkyl esters

3. 7-Acylami nocephalosporani c
acid

59

60

Absorption

Reference

Absorption

Oral

Property
Mod i f i ed

Oral

C--al koxycarbonyloxyalkyl esters


C,-ary1oxycarbony1oxyalkyl esters
C.-alkoxycarbonylami noalkyl esters

Route o f
Administration

C.-p-alkoxycarbonyloxybenzyl
esters

c.

b.

a.

Chemical
Modification

2. 7-Acylami nocephalosporani c
acid

1. 7-Acylcephalosporanic Acid

Parent Molecule

TABLE IV. CEPHALOSPORIN PRODRUGS DESIGNED TO MODIFY VARIOUS PROPERTIES


OF THE PARENT ANTIBIOTIC

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

7 - S u b s t i t u t e d cephalosporanic

S u b s t i t u t e d 7-3-aminocephemol- C . - p h y s i o l o g i c a l l y l a b i l e
4-carboxylic acid
esters

8.

9.

Oral

Oral

Oral

Oral

Route o f
Administration

^-Aminoacyloxymethyl e s t e r s

C.-acyloxymethyl e s t e r s

7-a-Amino (or ureido) c y c l o h e x a d i e n y l a l k y l cephalosporanic a c i d

7.

Imino ether

Chemical
Modification

7-Acy1 ami nocephalos porani c


acid

Molecule

(Continued)

6.

Parent

TABLE IV.

65

Absorption

67-69

Absorption,
i n c r e a s e d water
solubility,
66
decreased s i d e
effects

Absorption

6 4

Reference

Decreased
toxicity,
resistance to
3-lactamase

Property
Modified

140

PRO-DRUGS

Cephaloglycin
41 H CI (Acetoxy methyl ester)

- -

45 HCI (Pivaloyloxymethyl ester)

Journal of Antibiotics

Figure 1. Mean serum levels of cephaloglycin in fasting, healthy volunteers following oral administration of
200 mg of cephaloglycin and equimolar amounts of 41
HCI and 45 HCI (49)

Scheme XVII
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

2.

siNKULA

Pro-drug Approach to Antibiotics

141

For prodrug purposes, clindamycin can be c h e m i c a l l y modified


a t any o f the three hydroxyl groups present on the sugar p o r t i o n
o f the molecule.
Clindamycin A l k y ! E s t e r s . B i o l o g i c a l l y r e v e r s i b l e a l k y l
e s t e r s were sought t h a t would e l i m i n a t e the b i t t e r t a s t e o f
clindamycin by d e c r e a s i n g aqueous s o l u b i l i t y below t a s t e t h r e s h o l d
levels.
Chemistry. Standard s y n t h e t i c procedures were u t i l i z e d i n
the s y n t h e s i s o f 2- and 3-monoesters and 2,3-bis a l k y l e s t e r s .
(Schemes XVIII - XX). (81J.
Clindamycn-2-esters are s y n t h e s i z e d by u t i l i z i n g 3,4-0-pa n i s y l i d e n e - 7 ( S ) - c h l o r o - 7 - d e o x y l i n c o m y c i n 50 i n which the 3 and
4 hydroxyl groups are blocke
i s e s t e r i f i e d with e i t h e
a l k y anhydride,
o r an a l k y l chloroformate and the e s t e r - a c e t a l hydrolyzed i n
a c i d i c media to the 2-acyl e s t e r o f c l i n d a m y c i n .
S e l e c t i v e e s t e r i f i c a t i o n o f the 3 hydroxyl group i s achieved
by the use o f a low temperature r e a c t i o n medium ( p y r i d i n e a t -25
C) and a two-fold excess o f a l k y l chloroformate.
The 2,3-bis e s t e r s are made by treatment o f 49 with excess
a c y l a t i n g agent and using to advantage the stereochemistry o f
the hydroxyl groups. The C2 and C3 hydroxyl groups are e q u a t o r i a l
and r e l a t i v e l y c h e m i c a l l y r e a c t i v e whereas the C- hydroxyl i s
a x i a l and i s not e s t e r i f i e d to any a p p r e c i a b l e extent under these
conditions.
In v i t r o and i n v i v o s t u d i e s . Table V l i s t s p r e l i m i n a r y
a n t i b a c t e r i a l a c t i v i t i e s o f s e l e c t e d clindamycin e s t e r s using the
mouse as the model t e s t system (81J. In v i t r o data f o r a l l e s t e r
d e r i v a t i v e s show a c t i v i t y l e s s than c l i n d a m y c i n . T h i s appears
reasonable s i n c e many other e s t e r i f i e d a n t i b i o t i c s have been
shown to be i n a c t i v e u n t i l h y d r o l y s i s occurs (82-83).
The i n v i v o CD50 data i n mice i n d i c a t e t h a t v i r t u a l l y none
o f the e s t e r s possess a c t i v i t y comparable to c l i n d a m y c i n . Subsequent blood l e v e l s t u d i e s i n dogs, however, provided ample
evidence t h a t s e v e r a l 2-acyl e s t e r s were e q u i v a l e n t i n a c t i v i t y
(as measured by serum c o n c e n t r a t i o n o f clindamycin r e l e a s e d from
e s t e r ) to clindamycin HCI (81). T h i s d i f f e r e n c e i n b i o a c t i v i t y
between two animal s p e c i e s i l l u s t r a t e s the importance o f t e s t i n g
prodrug a n t i b i o t i c s i n s e v e r a l s p e c i e s before a d e c i s i o n i s
made to e l i m i n a t e a p o t e n t i a l candidate from f u r t h e r t e s t i n g
i n other animal s p e c i e s o r i n man.
Clindamycin-2-palmitate and -2-hexadecylcarbonate were
s e l e c t e d f o r comparative b i o a v a i l a b i l i t y s t u d i e s because (1)
f o l l o w i n g o r a l a d m i n i s t r a t i o n to dogs, each e s t e r produced serum
clindamycin l e v e l s e q u i v a l e n t to those obtained with clindamycin
HCI and (2) the e s t e r s lacked the c h a r a c t e r i s t i c b i t t e r n e s s o f
clindamycin.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

142

PRO-DRUGS

Scheme XVIII.

Clindamycin-2-monoesters

CH
'ISh

49

II
ROC C I ,
Base
-25C

CH

H-C-CI
I
C-NH-CH

S
ROC

HO^o
SCH
OH

Scheme XIX. Clindamycin-3-monocarbonate esters

Acylation
Pyridine.R.T.

Scheme XX. Clindamycin^,3-biscarbonate esters


In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.
5Q

690 (69)
2-hexanoate h y d r o c h l o r i d e
330 (33)
2-hexylcarbonate h y d r o c h l o r i d e
20 (2.0)
2-laurate hydrochloride
21 (2.1)
2-palmitate h y d r o c h l o r i d e
<4 (<0.4)
2-hexadecylcarbonate h y d r o c h l o r i d e
55 (5.5)
2-(p-benzoyl) benzoate h y d r o c h l o r i d e
7 (0.7)
2-(o-benzoyl) benzoate h y d r o c h l o r i d e
180
(18)
3-pentylcarbonate h y d r o c h l o r i d e
2,3-bis (hexylcarbonate) h y d r o c h l o r i d e <4 (<0.4)

0.67
1.31
1.36
0.43
<0.28
<0.1
<0.3
0.21
<0.1
0.23
1.67
0.54
<0.15
0.29
0.37
0.67
0.60
0.18
JOURNAL OF PHARMACEUTICAL

SCIENCES

a c t i v i t i e s c a l c u l a t e d as c l i n d a m y c i n base e q u i v a l e n t s . A s measured on a standard curve agar assay


versus S a r c i n a l u t e a . Results expressed as micrograms o f clindamycin base a c t i v i t y per m i l l i g r a m o f
e s t e r and as percent o f l i n c o m y c i n base a c t i v i t y ( i n p a r e n t h e s i s ) . Median p r o t e c t i v e dose r e l a t i v e
to t h a t o f c l i n d a m y c i n (clindamycin = 1.0) i n the mouse. (81_).

Clindamycin
Clindamycin
Clindamycin
Clindamycin
Clindamycin
Clindamycin
Clindamycin
Clindamycin
Clindamycin

In V i t r o
A c t i v i t y , . R e l a t i v e Median P r o t e c t i v e Dose ( C D )
mcg./mg.
Subcutaneous
Oral

Antibacterial Activity

TABLE V. ANTIBACTERIAL ACTIVITY OF CLINDAMYCIN 2- and 3-M0N0ESTERS AND 2,3-DIESTERS

144

PRO-DRUGS

Following a study o f t h e two e s t e r s and clindamycin HCI i n


twelve healthy male v o l u n t e e r s , F o r i s t e t . a l . (84) f i t t e d t h e
r e s u l t i n g serum c o n c e n t r a t i o n data t o a one-compartment open
pharmacokinetic model.
Clindamycin
i n serum
r e p r e s e n t s the f i r s t - o r d e r r a t e constant f o r appearance o f
clindamycin i n the serum. Both h y d r o l y s i s and a b s o r p t i o n a r e
represented i n t h i s constant f o r the e s t e r s . KE represents t h e
f i r s t - o r d e r r a t e constant f o r e l i m i n a t i o n o f clindamycin from the
body. Other parameters
included h a l f - l i v e s f o
b i o a c t i v i t y , serum c o n c e n t r a t i o n maximum and time o f maximum
c o n c e n t r a t i o n as well as area under t h e c o n c e n t r a t i o n vs. time
curve (0
>>) and a r e l i s t e d i n Table V I .
The r a t e o f appearance (A^) o f clindamycin and the p a l m i t a t e
e s t e r i n serum a r e not s i g n i f i c a n t l y d i f f e r e n t from each other
but both were more r a p i d l y absorbed than the hexadecylcarbonate
e s t e r . I t appears t h a t clindamycin-2-hexadecylcarbonate may not
be hydrolyzed as r a p i d l y o r as completely as the p a l m i t a t e e s t e r .
T h i s o b s e r v a t i o n i s f u r t h e r s u b s t a n t i a t e d by a comparison o f t h e
serum c o n c e n t r a t i o n maxima, the estimated time o f maximum concen
t r a t i o n and t o t a l area under the c o n c e n t r a t i o n vs. time curve
f o r t h e two e s t e r s . Based on t h i s study, clindamycin-2-palmitate
became the candidate o f choice f o r f u r t h e r c l i n i c a l t r i a l s .
Subsequent i n v e s t i g a t i o n s on l a r g e r p a t i e n t populations have
v e r i f i e d the r e s u l t s o f t h i s twelve s u b j e c t study (85-89).
CIindamycin-2-phosphate.
The phosphate e s t e r o f clindamycin
was s y n t h e s i z e d t o provide a b i o r e v e r s i b l e form o f clindamycin
devoid o f pain and i r r i t a t i o n upon i n j e c t i o n . The i r r i t a t i o n
caused by i n j e c t a b l e clindamycin was thought t o a r i s e from pre
c i p i t a t i o n o f the f r e e base a t the s i t e o f i n j e c t i o n o r l y s i s
due t o drug p a r t i t i o n i n g i n t o c e l l s surrounding the i n j e c t i o n
s i t e . A t p h y s i o l o g i c a l pH (7.4)clindamycin HCI i s s o l u b l e
o n l y t o the extent o f 3 mg./ml. whereas clindamycin-2-phosphate
i s r e a d i l y s o l u b l e a t t h i s pH (>150 mg./ml.) and should not
p r e c i p i t a t e a t the i n j e c t i o n s i t e . F u r t h e r , p e n e t r a t i o n o f
clindamycin-2-phosphate i n t o t h e c e l l should be n e g l i g i b l e s i n c e
i t s l i p o p h i l i c c h a r a c t e r i s d r a s t i c a l l y diminished v i a the hydrophi l i e phosphate e s t e r .
Chemistry ( 9 0 , 9 ] j . The C3 and C4 hydroxyl groups o f c l i n
damycin a r e c h e m i c a l l y blocked by formation o f 3 , 4 - 0 - p - a n i s y l i dene-7(S)-chloro-7-deoxylincomycin 50. (Scheme XXI).
Treatment
o f 50 with cyanoethyl phosphate and d i c y c l o h e x y l c a r b o d i i m i d e (DCC)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

2.

siNKULA

145

Pro-drug Approach to Antibiotics

TABLE VI
Pharmacokinetic Parameters Estimated from Serum Clindamycin B i o a c t i v i t y Concentrations Following Oral A d m i n i s t r a t i o n (150 mg.)
o f Clindamycin-2-Palmitate ( A ) , Clindamycin-2-Hexadecylcarbonate
(B), and Clindamycin H y d r o c h l o r i d e (C)
Parameter

K. ( h r . " )

2.81(0.92)
1.23-4.38&

1.53(0.94)
0.75-3.41

7.65(7.56)
1.03-22.92

A, ( h r . )

0.28(0.11)
0.16-0.56
0.25

0.58(0.25)
0.20-0.93
0.76

0.19(0.18)
0.03-0.67
0.15

Estimated time o f
1.16(0.24)
maximum c o n c e n t r a t i o n ( h r . ) 0.86-1.62
1.15

1.67(0.32)
1.09-2.09
1.80

0.86(0.53)
0.19-1.88
0.78

Estimated maximum
c o n c e n t r a t i o n (yg/ml)

1.99(0.61)
1.29-3.34
1.97

1.47(0.48)
0.97-2.33
1.44

2.61(1.14)
1.75-5.75
2.38

Observed maximum
c o n c e n t r a t i o n (pg/ml)

2.05(0.66)
1.33-3.38
2.05

1.47(0.54)
0.96-2.39
1.42

2.80(1.26)
1.67-6.40
2.39

- 1

0.25(0.07)
0.17-0.35
0.24

0.35(0.19)
0.17-0.81
0.36

0.26(0.07)
0.18-0.35
0.23

E, ( h r . )
*

2.94(0.83)
2.00-4.08
2.83

2.48(1.14)
0.85-4.17
1.95

2.84(0.76)
2.00-3.93
3.04

(hr. )

Area, 0 ( p g

hr./ml)

10.68(4.04) 8.22(3.85) 12.33(4.37)


5.63-21.14
3.71-15.40
8.36-20.04
10.20
7.37
12.20
JOURNAL OF PHARMACOKINETICS AND BIOPHARMACEUTICS
Mean (SD) f o r 12 s u b j e c t s .
''Range.
c Average obtained from mean serum l e v e l s f o r 12 s u b j e c t s . (84).
a

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

146

PRO-DRUGS

y i e l d s the cyanoethyl phosphate e s t e r intermediate 51^ which i s


converted t o clindamycin-2-cyanoethyl
phosphate 52 a f t e r a c i d
treatment. The intermediate 51_ i s p u r i f i e d by r e c r y s t a l l i z a t i o n
from water. Base h y d r o l y s i s o f the phosphodiester with ammonium
hydroxide on a T E A E - c e l l u l o s e column packed with IN ammonium
acetate a f f o r d s clindamycin-2-phosphate 53.
An a l t e r n a t i v e procedure i n v o l v e s the treatment o f 50 with
P O C W p y r i d i n e . A d d i t i o n o f water y i e l d s the phosphomonoester
3,4-u-anisylidene a c e t a l 54. A c i d h y d r o l y s i s o f the a c e t a l leads
to good y i e l d s o f 53.
B i o a c t i v i t y . Clindamycin-2-phosphate a c t i v i t y i n v i t r o i s
very low (<1%) i n d i c a t i n g t h a t t h e phosphate e s t e r possesses
very l i t t l e , i f any, a n t i b a c t e r i a l a c t i v i t y per se (92) The
median p r o t e c t i v e dose
i n f e c t e d with Staphylococcu
clindamycin HCI (6.6 mg./kg. v s . 4.9 mg./kg.), r e f l e c t i n g a
slowed r a t e o f h y d r o l y s i s o f t h e b i o i n a c t i v e e s t e r . S i n g l e
subcutaneous doses o f 10 mg./kg. i n r a t s produce lower blood
l e v e l s f o r the phosphate v s . clindamycin again i n d i c a t i n g t h a t
the phosphate e s t e r experiences a s l i g h t delay i n h y d r o l y s i s
i n v i v o . This study i s summarized i n Table VII ( 9 2 ) .
C l i n i c a l s t u d i e s i n human volunteers reveal a p a t t e r n s i m i l a r
to t h a t found i n t h e lower animal s p e c i e s . Thus, DeHaan e t . a l . ,
(93) found t h a t the e s t e r i s absorbed i n t a c t a f t e r m u l t i p l e i n tramuscular i n j e c t i o n s but i s r a p i d l y hydrolyzed i n the serum
(Figure 2). In f a c t , a f t e r a t e n minute intravenous i n f u s i o n
of clindamycin phosphate, the mean h a l f - l i f e o f t h e e s t e r i n
serum i s estimated t o be 9.6 minutes (Figure 3) v e r i f y i n g the
r a p i d conversion o f b i o i n a c t i v e e s t e r t o clindamycin.
Other s t u d i e s i n animals (94) and i n man (95) have demons t r a t e d t h e l a c k o f l o c a l i r r i t a t i o n and pain with i n j e c t a b l e
clindamycin phosphate.
Summary
T h i s b r i e f review h i g h l i g h t s the importance o f the use o f an
i n t e r d i s c i p l i n a r y approach i n t h e s y n t h e s i s and t e s t i n g o f a n t i b i o t i c prodrug d e r i v a t i v e s . Along with the development o f unique
chemistry f o r the s y n t h e s i s o f such prodrugs, a knowledge and
a p p r e c i a t i o n o f the various i n v i t r o and i n v i v o methods o f
b i o l o g i c a l e v a l u a t i o n o f such d e r i v a t i v e s i s necessary t o r a t i o n a l l y choose those best s u i t e d f o r c l i n i c a l use. Further, s e l e c t e d
pharmacokinetic s t u d i e s a r e v a l u a b l e i n the determination o f
the a b s o r p t i o n , d i s t r i b u t i o n , metabolism, and e l i m i n a t i o n
c h a r a c t e r i s t i c s o f such drugs.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

' C a l c u l a t e d as molecular e q u i v a l e n t s o f clindamycin base.


W e a under time : c o n c e n t r a t i o n curve standard e r r o r s . Values r e p r e s e n t mean o f three determina
t i o n s with 3 r a t s per d e t e r m i n a t i o n .
W e a v a l u e s r e l a t i v e t o t h a t o f c l i n d a m y c i n . Clindamycin = 1.00.
H i m e (min) a t which 50% o f area i s under the time : c o n c e n t r a t i o n curve. ( 9 2 ) .

JOURNAL OF ANTIBIOTICS

1.23 mcg/ml a t 45 mi

74 5

0.76

136 32

Clindamycin-2-P0

2.25 mcg/ml a t 30 mi

72 10

1.00

177 30

Clindamycin

Mean time o f
Max. cone.(mcg/ml)
50% area 2 S E '
a t time t

Area under , x
curve 2 SE '
R e l a t i v e area
o f curves '

dose o f c l i n d a m y c i n and

Antibiotic

Comparative b i o a c t i v i t i e s i n whole blood o f r a t s a f t e r a s i n g l e subcutaneous


clindamycin 2-PCL a t 10 mg./kg. '.

TABLE VII

148

PRO-DRUGS

Scheme XXI

300mg q8h 43 doses

4
Hr. after 1st Dose

8j

7.5

2nd dose 3rd dose

| 0

43rd Dose
Journal of Clinical Pharmacology

Figure 2. Mean serum concentrations (pg/ml) of clindamycin and clindamycin pho


after I.M. injections of clindamycin phosphate (93)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

siNKULA

Pro-drug Approach to Antibiotics

20fl

18

Clindamycin bioactivity
"Total" clindamycin +
clindamycin phosphate)

1st dose; 10-min. infusion

\/

Estimated mean half-time for


hydrolysis in serum 9.6 minutes

/
/
/

/
-Total"
^Clindamycin phosphate

Clindamycin-

10

12

Hr. after Start of Infusion


Journal of Clinical Pharmacology

Figure 3. Mean serum concentrations (\*>g/m\) of clindamycin


and clindamycin phosphate after a 300-mg LV. infusion of
clindamycin phosphate (10-min infusion) (93)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

150

PRO-DRUGS

Acknowledgements
The author wishes to express his appreciation to Dr.
W. Morozowich for helpful suggestions concerning the synthesis
of clindamycin phosphate. Special thanks are also extended to
D. Harling for typing and to E.E. Beals for preparation of the
schemes and figures used throughout the manuscript.
Literature Cited

1. von Daehne, W., Frederiksen, E., Gunderson, E., Lund, F.,


Mrch, P., Petersen H.J. Roholt . Tybring L .
d
Godtfredsen, W.O.
2. von Daehne, W., Godtfredsen, W.O., Roholt, ., and Tybring,
L., Antimicrob. Ag. Chemother., (1970), 431.
3. Hardcastle, G.A., J r . , Johnson, D.A., Panetta, C.A., Scott,
A.I., and Sutherland, S.A., J. Org. Chem., (1966), 31, 897.
4. Johnson, D.A., and Panetta, C.A., U.S. Patent, 3,198,804
(August 3, 1965).
5. Sleezer, P.D., and Johnson, D.A., Belgian Patent 788,720
(March 12, 1973).
6. Essery, J.M., U.S. Patent 3,679,663 (July 25, 1972).
7. Schwartz, M.A., and Hayton, W.L., J. Pharm. Sci., (1972), 61,
906.
8. Jusko, W.J., and Lewis, G.P., Ibid., (1973), 62, 69.
9. Hobbs, D.C., Antimicrob. Ag. Chemother., (1972), 2, 272.
10. Butler, ., and Hamanaka, E.S., U.S. Patent 3,681,342 (August
1, 1972).
11. Brain, E.G., and Nayler, J.H.C., U.S. Patent 3,282,926
(November 1, 1966).
12. English, A.R., Retsema, J.., Ray, V.A., and Lynch, J.E.,
Antimicrob. Ag. Chemother., (1972), 1, 185.
13. Farbwerke Hoechst, Netherlands Patent 73,09128 (January 8,
1974).
14. Petersen, H.J., J. Med. Chem., (1974), 17, 101.
15. Frederiksen, E.K., and Godtfredsen, W.O., U.S. Patent 3,719,
668 (March 6, 1973).
16. Jensen, A.B.A., and Russell, T.J., J. Chem. Soc., (1965),
2127.
17. Hansson, E., Magni, L . , and Wahlgvist, S., Antimicrob. Ag.
Chemother., (1967), 568.
18. Ramsey, C . H . , Bodin, N.O., and Hansson, E., Arzneim. -Forsch.
(1972), 22, 1962.
19. Foltz, E.L., West, J.W., Breslow, I.H., and Wallick, H.,
Antimicrob. Ag. Chemother., (1970), 442.
20. Jordan, M.C., deMaine, J.B., and Kirby, W.M.M., Ibid., 438.
21. Hultberg, E.R., and Backelin, B., Scand. J. Infect. Dis.,
(1972), 4, 149.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

SINKULA

22.

Frederiksen, E . K . , and Godtfredsen, W.O., U.S. Patent 3,697,


507 (October 10, 1972).
Jusko, W . J . , Lewis, G . P . , and Schmitt, G.W., C l i n . Pharmacol.
Ther., (1973), 14, 90.
Yurchenco, J.., Hopper, M.W., Vince, T.D., and Warren, G . H . ,
Chemotherapy, (1972), 17, 405.
E.R. Squibb and Sons, Inc., West German Patent 2,152,745
(April 27, 1972).
Essery, J.M., and West, J.R., U.S. Patent 3,654,265
(April 4, 1972).
Farbwerke Hoechst AG, Netherlands Patent 7,309,129 (January
8, 1974).
Sellstedt, J.H., and Wolf, ., U.S. Patent 3,692,774
(September 19, 1972)
Andreu, SA, West
Merck and C o . , Inc., West German Patent 2,256,538 (May 24,
1973).
Farbwerke Hoechst AG, West German Patent 2,159,555 (June 7,
1973).
Beecham Group L t d . , Belgian Patent 784,698 (December 11,
1972).
Soc. Rech. et Applic. Ind., French Patent 2,181,505
(December 7, 1973).
Soc. Rech. et Applic. Ind., French Patent 2,181,506
(December 7, 1973).
Boniece, W.S., Wick, W.E., Holmes, D . H . , and Redman, C.E.,
J. Bacteriol.,(1962), 84, 1292.
Naumann, P . , and Fedder, J., Int. J. C l i n . Pharmacol.
Beiheft Oracef., (1970), 6.
Wick, W.E., in "Cephalosporins and Penicillins - Chemistry
and Biology", E.H. Flynn, e d . , pp. 515-517, Academic Press,
New York, NY, 1972.
Wick, W.E., and Boniece, W.S., Appl. Microbiol., (1965), 13,
243.
Wick, W.E., I b i d . , (1967), 15, 765.
Applestein, J.M., Crosby, E.B., Johnson, W.D., and Kaye, D.,
Ibid., (1968), 16, 1006.
Thornhill, T.S., Levison, M.E., Johnson, W.D., and Kaye, D . ,
Ibid., (1969), 17, 457.
Flynn, E . H . , U.S. Patent 3,218,318 (November 16, 1965).
Kukolja, S . , U.S. Patent 3,728,342 (April 17, 1973).
Kukolja, S . , J. Med. Chem., (1970), 13, 1114.
Jensen, E.F., Jang, R . , and MacDonnell,L.R.,Arch. Biochem.,
(1947), 15, 415.
Van Heyningen, E., J. Med. Chem., (1965), 8, 22.
Chauvette, R.R., and Flynn, E . H . , J. Med. Chem., (1966), 9,
741.
Nefkens, G . H . L . , Tesser, G.I., and Nivard, R.J.F., Rec. Trav.
Chim., (1962), 81, 683.

23.
24.
25.
26.
27.
28.
29.
30.
31.
32.
33.
34.
35.
36.
37.
38.
39.
40.
41.
42.
43.
44.
45.
46.
47.
48.

Pro-drug Approach to Antibiotics

151

2.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

152

PROD
- RUGS

49. Binderup, E., Godtfredsen, W.O., and Roholt, ., J. Antibiot.


(1971), 24, 767.
50. Cheney, L . C . , Godfrey, J.C., Crast, L.B., Jr., and Luttinger,
J.R., U.S. Patent 3,284,451 (November 6, 1966).
51. Webber, J.., Huffman, G.W., Koehler, R . E . , Murphy, C.F.,
Ryan, C.W., Van Heyningen, E.M., and Vasileff, R.T., J. Med.
Chem., (1971), 14, 113.
52. Jen, T., Dienel, B . , Frazee, J., and Weisbach, J., J. Med.
Chem., (1972), 15, 1172.
53. Webber, J.., and Van Heyningen, E.M., U.S. Patent 3,708,
480 (January 2, 1973).
54. Wick, W.E., Antimicrob. Ag. Chemother., (1966), 870.
55. Lee, C.C., Herr, E . B . , and Anderson, R.C., Clin. Med., (1963)
70, 1123.
56. Chauvette, R.R., Flynn
Morin, R.B., Mueller, R.A., Pioch, R.P., Roeske, R.W., Ryan,
C.W., Spencer, J.L., and Van Heyningen, E., Antimicrob. Ag.
Chemother., (1963), 687.
57. Sullivan, H.R., Billings, R . E . , and McMahon, R.E., J . Anti
biot., (1969), 22, 27.
58. Wick, W.E., Wright, W.E., and Kuder, H.V., Appl. Microbiol.,
(1971), 21, 426.
59. Astra Lakemedel AB, Netherlands Patent 73,03437 (September
17, 1973).
60. Yamanouchi Pharmaceutical Co. Ltd., Japanese Patent 4,720,
187 (September 27, 1972).
61. Yamanouchi Pharmaceutical Co. Ltd., Belgian Patent 781,659
(July 31, 1972).
62. Yamanouchi Pharmaceutical Co. Ltd., West German Patent 2,223,
588 (August 9, 1973).
63. Merck and Co., Inc., Netherlands Patent 73,09893 (February
5, 1974).
64. Glaxo Labs, Ltd., West German Patent 2,223,375 (November 23,
1972).
65. E.R. Squibb and Sons, Inc., West German Patent 2,152,745
(April 27, 1972).
66. Loevens Kemiske Fab Produktions A/S, West German Patent
2,230,620 (December 28, 1972).
67. Ciba-Geigy AG, Netherlands Patent 73,09136 (January 2, 1974).
68. Ciba-Geigy AG, Netherlands Patent 73,09137 (January 2, 1974).
69. Ciba-Geigy AG, Netherlands Patent 73,09139 (January 2, 1974).
70. Maggi, N., Gallo, G.G., and Sensi, P., Farmaco. Ed. Sci.,
(1967), 22, 316.
71. Maggi, N . , Pallanza, R., and Sensi, P., Antimicrob. Ag.
Chemother., (1965), 765.
72. Furesz, S., Arioli, V . , and Pallanza, R., Ibid., (1965), 770.
73. Sensi, P.,Maggi, N . , Furesz, S., and Maffii, G . , Ibid.,
(1966), 699.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

2. sN
i KULA
74.
75.
76.
77.
78.
79.
80.
81.
82.
83.
84.
85.
86.
87.
88.
89.
90.
91.
92.
93.
94.
95.

Pro-drug Approach to Antibiotics

Furesz, S., Scotti, R., Pallanza, R., and Mapelli, E.,


Arzneim. -Forsch., (1967), 17, 726.
Acocella, G . , Nicolis, F . B . , and Lamarina, ., "A Study on
the Kinetics of Rifampicin in Man", Vth International
Congress of Chemotherapy, Vienna, 1967.
Maggi, . , Vigevani, . , and Pallanza, R., Experientia,
(1968), 24, 209.
Maggi, . , Furesz, S., Pallanza, R., and Pelizza, G . , Arz
neim.-Forsch, (1969), 191, 651.
Wagner, J.G., Novak, E., Patel, N.C. Chidester, C . G . ,
and Lummis, W.L., Amer. J. Med. Sci., (1968), 256, 25.
McGehee, R.F., Jr., Smith, C.B., Wilcox, C., and Finland,
M., Ibid., 279.
DeHaan, R.M., Metzler C.M., Schellenberg D. VandenBosch
W.D., and Masson,
Toxicol., (1972), ,
Sinkula, .., Morozowich, W., and Rowe, E.L., J. Pharm. Sci.
(1973), 62, 1106.
Wick, W.E., and Mallett, G . E . , Antimicrob. Ag. Chemother.,
(1968), 410.
Tardrew, P.L., Mao, J.C.H., and Kenney, D., Appl. Microbiol.,
(1969), 58, 2140.
Forist, .., DeHaan, R.M., and Metzler, C.M., J. Pharmaco
kinet. and Biopharmaceutics, (1973), 1, 89.

DeHaan, R.M., VandenBosch, W.D., and Metzler, C.M., J. Clin.


Pharmacol., (1972), 12, 205.
DeHaan, R.M., and Schellenberg, D., Ibid., (1972), 12, 74.
DeHaan, R.M., Schellenberg, D., VandenBosch, W.D., and Maile,
M.H., Curr. Ther. Res., (1972), 14, 81.
Pfeifer, R.T., DeHaan, R.M., VandenBosch, W.D., and Schellen
berg, D., Clin. Med., (1973), 80, 21.
Metzler, C.M., DeHaan, R., Schellenberg, D., and VandenBosch,
W.D., J . Pharm. Sci., (1973), 62, 591.
Morozowich, W., Lamb, D.J., DeHaan, R.M., and Gray, J.E.,
Abstracts of Papers, APhA Academy of Pharmaceutical Sciences,
Washington, D.C. Meeting, April 1970, p. 63.
Morozowich, W., and Lamb, D.J., U.S. Patent 3,487,068
(December 30, 1969).
Brodasky, T.F., and Lewis, C., J. Antibiot., (1972), 25,
230.
DeHaan, R.M., Metzler, C.M., Schellenberg, D., and Vanden
Bosch, W.D., J. Clin. Pharmacol., (1973), 13, 190.
Gray, J.E., Weaver, R.N., Moran, J., and Feenstra, E.S.,
Toxicol. Appl. Pharmacol., (1974), 27, 308.
Edmondson, H.T., Ann. Surg., (1973), 178, 637.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

153

3
The Chemistry of a Novel 5,5-Diphenylhydantoin
Pro-drug
V. STELLA, T. HIGUCHI, A. HUSSAIN, and J. TRUELOVE
University of Kansas, Department of Pharmaceutical Chemistry and
INTERx Research Corp., Lawrence, Kans. 66044
Phenytoin or 5,5-diphenylhydantoin (DPH), a
widely used anticonvulsan
primaril i
the treatment of gran
erratic absorption and dissolution patterns as well
as possible precipitation in the body after intravenous (I.V.) infusions of highly alkaline solutions
of its sodium salt (1-6). It was the objective of
this work to find a water soluble pro-drug (7,8)
form of DPH which would revert to DPH in the body.
The aim was to improve both the parenteral and oral
bioavailability of DPH.
A number of hydantoins including DPH, mephenytoin
or 5-ethyl-3-methyl-5-phenylhydantoin (I), and nitrofurantoin or 1-(5-nitro-2-furfurylindene) aminohydantoin (II) are widely used as drugs. Their uses,
however, have been hampered by their low water solubility combined with their weakly acidic nature.
Orally the bioavailability of DPH from capsules
has been erratic (1-3). The P a of DPH, approximately 8.3 (1,9), allows DPH to be formulated as its
mono-sodium salt. Formulations of DPH as the free
acid are also available. A report before a congressional investigation committee in 1967 suggested
that a brand change of DPH resulted in increased convulsive seizures due to lower bioavailability from
the new formulation. Similarly differences between
products have been noted by Martin et al. (2) and by
Arnold et al. (1). The dependency of DPH blood levels
after oral dosing on differences, in the rate of
metabolism of DPH, particle size, and various generic
and trade name products has recently been discussed
by Glazko (10). DPH has an aqueous solubility of
between 1-4 mg/100 ml (1)and it appears that this
poor aqueous solubility is the major cause of the
erratic absorption and dissolution rates (1) of
K

154
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

STELLA

E T

AL.

Chemistry of a 5,5-Diphenylhydantoin Pro-drug

(ID

v a r i o u s DPH p r e p a r a t i o n s .
The low aqueous s o l u b i l i t y o f DPH can be a t t r i b u t e d t o i t s s t r o n g c r y s t a l
l a t t i c e (mp. 293) r e s u l t i n g from i n t e r m o l e c u l a r h y d r o gen b o n d i n g (11).
That the s t r o n g c r y s t a l l a t t i c e i s
a major f a c t o r i n the d e t e r m i n a t i o n o f s o l u b i l i t y i s
a l s o e x e m p l i f i e d by the poor l i p i d s o l u b i l i t y o f DPH.
DPH i s g i v e n p a r e n t e r a l l y as i t s mono-sodium s a l t
in a reconstituted i n j e c t i o n with a solvent consist i n g o f "40% p r o p y l e n e g l y c o l and 10% a l c o h o l i n water
f o r i n j e c t i o n and i s b u f f e r e d t o pH 10 t o 12 w i t h
sodium h y d r o x i d e " (12) .
Blum e t a l . (_5,6) and o t h e r s
(13) n o t e d s e v e r e s i d e e f f e c t s " T h dogs and man when
DPH i n j e c t i o n s were g i v e n i n t r a v e n o u s l y .
A subsequent
autopsy o f the dogs used i n the experiments o f Blum
e t a l . (5^,6) showed p r e c i p i t a t i o n o f DPH i n the l u n g s ,
I T e . , the i n j e c t i o n o f sodium DPH o f pH 12 on m i x i n g
w i t h b l o o d b u f f e r e d a t pH 7.4 r e s u l t e d i n the p r e c i p i t a t i o n o f DPH as the f r e e a c i d .
Precautions i n
the I . V . use o f i n j e c t a b l e DPH i n the t r e a t m e n t o f
s t a t u s e p i l e p t i c u s and d i g i t a l i s i n t o x i c a t i o n
(13)
and i n the attempted p r e p a r a t i o n o f l a r g e volume I . V .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

155

156

PRO-DRUGS

i n j e c t i o n s o f DPH have r e c e n t l y been t h e c e n t e r


of d i s c u s s i o n (14-19).
The use o f sodium DPH v i a t h e i n t r a m u s c u l a r
(I.M.)
r o u t e has a l s o been c r i t i c i z e d ( 2 0 - 2 3 ) .
Blood l e v e l s
o f DPH a f t e r I . M . a d m i n i s t r a t i o n were g e n e r a l l y found
t o be lower than t h o s e from o r a l d o s i n g .
Muscle d i s s e c t i o n two days a f t e r sodium DPH was g i v e n I . M . t o
r a b b i t s showed t h a t almost 50% o f t h e a d m i n i s t e r e d
dose remained a t t h e s i t e o f i n j e c t i o n .
DPH had
o b v i o u s l y p r e c i p i t a t e d a t the s i t e o f t h e i n j e c t i o n
(21).
C u r r e n t dosage forms o f DPH f o r o r a l a b s o r p t i o n
r e s u l t i n f a i r l y complete b i o a v a i l a b i l i t y and p a t i e n t
t o p a t i e n t v a r i a b i l i t y c a n be a t t r i b u t e d t o d i f ferences i n metabolis
tion.
The v e r y e r r a t i c b l o o d l e v e l s and o c c a s i o n a l
t o x i c i t y o f DPH i n c h i l d r e n (24,25) cannot seem t o
be c o m p l e t e l y a s c r i b e d t o m e t a b o l i s m .
After oral
d o s i n g the b l o o d l e v e l maximum o f DPH c a n o c c u r 6-12
hours a f t e r d o s i n g , s u g g e s t i n g t h a t a b s o r p t i o n t a k e s
p l a c e a l o n g t h e whole o f t h e g a s t r o i n t e s t i n a l (GI)
tract.
The completeness o f a b s o r p t i o n i n a m a r g i n a l l y
b i o a v a i l a b l e p r o d u c t w i l l o b v i o u s l y be a f f e c t e d by
the r e s i d e n t time o f t h e d r u g i n t h e GI t r a c t , i . e . ,
t r a n s i t time.
The s h o r t e r r e s i d e n t time noted i n
c h i l d r e n may account f o r t h e more e r r a t i c b e h a v i o r
o f DPH i n c h i l d r e n r e l a t i v e t o a d u l t s .
As p r e v i o u s l y s t a t e d , t h e aim o f t h i s i n v e s t i g a t i o n was t o determine t h e f e a s i b i l i t y o f o b t a i n i n g
transient pro-drug
forms o f DPH w h i c h would
c o n f e r a c c e p t a b l y h i g h e r aqueous s o l u b i l i t y w i t h i n
the p h y s i o l o g i c a l l y c o m p a t i b l e pH range b u t once
i n t h e body r e v e r t t o DPH i n a r e l a t i v e l y s h o r t
period of time.
The o n l y p r e v i o u s a t t e m p t s a t
p r o - d r u g forms o f DPH appeared i n the work o f Nakamura
e t a l . (26-29) who showed t h a t
3-carbethoxy-5,5-dip h e n y l h y d a n t o i n ( I I I ) was a l e s s e r r a t i c b i o a v a i l a b l e

N-C-OC H .

HN

XT

(III)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

STELLA

E T

Chemistry of a 5,5-Diphenylhydantoin Pro-drug

AL.

form o f DPH when compared t o DPH i t s e l f .


Ill,
however, was n o t water s o l u b l e so c o u l d n o t be
g i v e n by I . V . i n j e c t i o n .
3-N , -Diethylaminoethyl-2-ethyl-5-methyl2 - p h e n y l h y d a n t o a t e (IV) was r e c e n t l y s y n t h e s i z e d
and i t s u s e f u l n e s s as a water s o l u b l e p r o - d r u g o f
mephenytoin (30) was shown.
1

/ 2 5
OC H -N-H
C

NHCNHC
I
CH

C H
2

(IV)
pH = 7 . 4 , = 37<
t i = 20 m i n .

C H -C
2

(I)
The s y n t h e s i s , c h e m i s t r y , p h y s i c a l p r o p e r t i e s
and f o r m u l a t i o n problems o f a s i m i l a r d e r i v a t i v e t o
IV b u t f o r DPH w i l l be p r e s e n t e d a t t h i s t i m e .
Rationale
E s t e r s o f h y d a n t o i c a c i d s were known t o c y c l i z e
v i a an i n t r a m o l e c u l a r r e a c t i o n under b a s i c c o n d i t i o n s
t o t h e i r r e s p e c t i v e h y d a n t o i n (scheme I) b u t no
a c t u a l k i n e t i c d a t a was a v a i l a b l e .
K i n e t i c d a t a was
a v a i l a b l e on the c y c l i z a t i o n o f e s t e r s o f the r e l a t e d
O - u r e i d o b e n z o i c a c i d i n water i n the n e u t r a l t o a l k a
l i n e pH range (31).
I n t r a m o l e c u l a r c y c l i z a t i o n s had
been p o s t u l a t e d as a means o f l a t e n t i a t i o n by L e v i n e

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

157

158

PRO-DRUGS

R
OH

HN^
CNH

R"
I
C1

Il
4
-COR^

2 I
R-C-

HN

NCH-

Scheme I
e t a l . (32) i n t h e i r work on the c o n v e r s i o n s o f - h a l o alkylamines to t h e i
many d e r i v a t i v e s o f drugs formed t h r o u g h an e s t e r l i n k
age have been dependent on e n z y m a t i c r e a c t i o n s t o r e
l e a s e the p a r e n t compound (3J3,_34) .
The c y c l i z a t i o n s o f
the e s t e r s o f h y d a n t o i c a c i d a r e i n t r a m o l e c u l a r r e a c
t i o n s and t h e s e are known t o be many o r d e r s o f m a g n i
tude f a s t e r than t h e i r i n t e r m o l e c u l a r e q u i v a l e n t .
This
forms a b a s i s f o r the f o r m a t i o n o f the p a r e n t drug from
i t s p r o - d r u g which i s n o t dependent on e n z y m a t i c m e d i a
tion.
P r o c e d u r e and R e s u l t s
Because o f the a p p a r e n t s u c c e s s o f compound IV i n
p r o d u c i n g a water s o l u b l e p r o - d r u g o f I a s i m i l a r d e
r i v a t i v e t o IV b u t f o r DPH was s y n t h e s i z e d .
The com
pound o f i n t e r e s t was V .

(V)
The s y n t h e s i s o f V was a c c o m p l i s h e d v i a the
f o l l o w i n g p r o c e d u r e and pathway.
The f i r s t s t e p i n
v o l v e d the r e a c t i o n o f 2 , 2 - d i p h e n y l g l y c i n e w i t h e t h y l c h l o r o f o r m a t e under aqueous a l k a l i n e c o n d i t i o n s .
Ini
t i a l l y the s y n t h e s i s o f 2 , 2 - d i p h e n y l g l y c i n e was accom
p l i s h e d by t h e method o f D u s c h i n s k y (35.) which i n v o l v e d
the a c t u a l h y d r o l y s i s o f DPH i n 20% sodium h y d r o x i d e

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

STELLA

E T A L .

Chemistry of a 5,5-Diphenylhydantoin Pro-drug

i n an a u t o c l a v e under a n i t r o g e n atmosphere a t 1801 9 0 f o r 24 h o u r s .


The s y n t h e s i s o f 2 , 2 - d i p h e n y l g l y c i n e v i a o t h e r p r o c e d u r e s such as t h o s e found i n
O r g a n i c S y n t h e s i s , C o l l . v o l . 3, p p . 8 8 - 9 1 , 1965 met
with only p a r t i a l success.
The i n e x p e n s i v e and r e l a
t i v e l y s i m p l e s y n t h e s i s o f DPH meant t h a t i t was used
as a p r i m a r y s o u r c e o f 2 , 2 - d i p h e n y l g l y c i n e .
2,2D i p h e n y l g l y c i n e was s u b s e q u e n t l y o b t a i n e d from a com
m e r c i a l s o u r c e ( J . T . Baker and C o . ) which a p p a r e n t l y
a l s o used t h e h y d r o l y s i s o f DPH as i t s method o f
preparation.
Step I .
2 , 2 - D i p h e n y l g l y c i n e ( V I ) , 68.1 g (0.3
m o l e s ) , was p l a c e d i n a 2 l i t e r e r l e n m e y e r f l a s k and
d i s s o l v e d i n 700 ml 1 NaOH.
While s t i r r i n g v i g o r
o u s l y , 25 ml o f e t h y l c h l o r o f o r m a t
o f water was a d d e d .
The r e a c t i o n m i x t u r e was m a i n
t a i n e d a t a temperature o f 2 0 - 2 5 by p e r i o d i c a l l y
submerging i n an i c e b a t h .
The pH o f the r e a c t i o n
m i x t u r e was m a i n t a i n e d a t 11 o r above by p e r i o d i c
a d d i t i o n s o f NaOH p e l l e t s .
A f t e r 25-30 m i n u t e s ,
a n o t h e r 25 ml o f V I I and 100 ml o f H 0 was a g a i n
added and t h e pH c o n t i n u o u s l y a d j u s t e d w i t h NaOH
pellets.
T h i s p r o c e s s was r e p e a t e d u n t i l 250 ml
(283.8 g , 2.6 moles) o f V I I had been a d d e d . The
r e a c t i o n m i x t u r e was s t i r r e d an a d d i t i o n a l two hours
( t o t a l r e a c t i o n time ^6% hours) a t room t e m p e r a t u r e .
The p r o d u c t , V I I I , o f the r e a c t i o n was then i s o l a t e d
as f o l l o w s :
The r e a c t i o n m i x t u r e was e x t r a c t e d t w i c e w i t h
e t h e r ( f i r s t w i t h a volume o f e t h e r e q u a l t o t h e
volume o f the aqueous r e a c t i o n m i x t u r e and second
w i t h o n e - h a l f t h e volume o f t h e aqueous r e a c t i o n m i x
ture) .
The aqueous l a y e r was t r a n s f e r e d t o a l a r g e
beaker and s l o w l y a c i d i f i e d w i t h c o n c e n t r a t e d HCI
(NOTE;
The a c i d must be added s l o w l y t o p r e v e n t e x
c e s s foaming and l o s s o f t h e p r o d u c t . ) t o a pH o f
0 to 1.
At t h i s point, considerable s o l i d material
separated.
The a c i d i f i e d aqueous m i x t u r e was then e x t r a c t e d
w i t h e t h e r i n t h e same manner as a b o v e .
The e t h e r
l a y e r s were d r i e d o v e r anhydrous sodium s u l f a t e ,
f i l t e r e d , and e v a p o r a t e d w i t h t h e a i d o f a r o t a r y
evaporator to y i e l d a c l e a r , l i g h t - y e l l o w c o l o r e d ,
and v i s c o u s o i l . The o i l s l o w l y (1-2 hours) c r y s
t a l l i z e d upon s t a n d i n g a t room t e m p e r a t u r e .
The
c r y s t a l s were suspended i n p e t r o l e u m e t h e r , f i l t e r e d ,
and washed w i t h p e t r o l e u m e t h e r .
The r e a c t i o n i s o u t l i n e d on t h e f o l l o w i n g page
and u s u a l l y r e s u l t e d i n a p p r o x i m a t e l y 75% o f t h e
of the t h e o r e t i c a l y i e l d .
Compound V I I I o r
2

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

159

160

PRO-DRUGS

N - c a r b e t h o x y - 2 , 2 - d i p h e n y l g l y c i n e was found t o have


a melting p o i n t of 1 5 0 - 1 5 3 .
Step I I .
V I I I , 60 g (0.2 moles) was p l a c e d
i n a round-bottomed f l a s k f i t t e d w i t h a magnetic
s t i r r e r and r e f l u x condenser and 200 ml (328 g ,
7.75 moles) o f c l e a r , c o l o r l e s s t h i o n y l c h l o r i d e was
added.
The m i x t u r e was s l o w l y warmed t o a p p r o x i m a t e l y
8 0 and a l l o w e d to r e f l u x f o r one h o u r .
A f t e r the r e a c t i o n m i x t u r e was c o o l e d , the exc e s s t h i o n y l c h l o r i d e was removed r e s u l t i n g i n a
w h i t e to l i g h t y e l l o w s o l i d ( I X )
IX was t h o r o u g h l y
washed w i t h p e t r o l e u m e t h e r u n t i l a l l t r a c e s o f
y e l l o w c o l o r and t h i o n y l c l o r i d e odor were removed.
The r e a c t i o n sequence i s o u t l i n e d on the
f o l l o w i n g page and u s u a l l y r e s u l t s i n 95-100% o f the
theoretical yield:
The m e l t i n g p o i n t o f 4 , 4 - d i p h e n y l 2 , 5 - o x a z o l i d i n e d i o n e (IX) was found t o be 1 6 5 - 6 .
Step I I I .
I X , 50.6 g (0.2 m o l e s ) , was p l a c e d
i n a 500 ml 3-necked d i s t i l l i n g f l a s k f i t t e d w i t h
a s o u r c e o f HCI g a s , an exhaust t u b e , a magnetic
s t i r r e r , and an o i l b a t h as shown i n the s k e t c h on
the f o l l o w i n g page.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

Chemistry of a 5,5-Diphenylhydantoin Pro-drug 161

STELLA ET A L .

V-cCOOH

c=o

HN
C0C Hc
o

(VIII)

To HCI Trap

HCI

(gas)

Oil

Bath

Magnetic
Heater

Stirrer/

A f t e r the a d d i t i o n o f I X , the f l a s k was


f l u s h e d w i t h HCI (gas) and 27.5 ml (24.6 g ,
0.21
moles) o f , - d i e t h y l a m i n o e t h a n o l was added.
The
s t o p p e r was r e p l a c e d and the HCI (gas) f l o w r e s t a r t e d
immediately.
The m i x t u r e was then lowered i n t o an
o i l b a t h a t 9 8 - 1 0 5 , and a l l o w e d t o r e a c t f o r 15
minutes under a c o n t i n u o u s b l a n k e t o f HCI g a s .
At
the end o f 15 m i n u t e s , e v o l u t i o n o f carbon d i o x i d e
c e a s e d and a c l e a r o i l y l i q u i d r e s u l t e d .
The f l a s k was removed from the o i l b a t h and a l
lowed t o c o o l u n t i l the r e a c t i o n m i x t u r e temperature
was below 6 0 .
The c o o l e d r e a c t i o n m i x t u r e was d i s s o l v e d i n
250 ml c h l o r o f o r m and f i l t e r e d .
A white c r y s t a l l i n e
m a t e r i a l i d e n t i f i e d as the d i h y d r o c h l r o i d e o f - ' , ' d i e t h y l a m i n o e t h y l - 2 , 2 - d i p h e n y l g l y c i n a t e (X) was

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

162

PRO-DRUGS

o c c a s i o n a l l y observed at t h i s p o i n t .
This material
may be saved by washing t h o r o u g h l y w i t h c h l o r o f o r m ,
f i l t e r i n g , and a d d i n g i t to the c h l o r o f o r m s o l u t i o n
o f X i m m e d i a t e l y p r i o r t o the NaOH e x t r a c t i o n
procedure below.
The c h l o r o f o r m f i l t r a t e was p l a c e d on a r o t a r y
e v a p o r a t o r and a p p r o x i m a t e l y 50 ml o f c h l o r o f o r m were
removed.
Any e x c e s s HCI d i s s o l v e d i n the r e a c t i o n
m i x t u r e was removed by t h i s p r o c e d u r e .
The r e
m a i n i n g c h l o r o f o r m " r e a c t i o n m i x t u r e " s o l u t i o n was
then e x t r a c t e d w i t h aqueous NaOH u n t i l the aqueous
l a y e r had a pH > 11.
Each f r a c t i o n (aqueous and o r
g a n i c ) was b a c k - e x t r a c t e d and the c h l o r o f o r m l a y e r s
combined w i t h anhydrous sodium s u l f a t e .
A f t e r d r y i n g , th
c h l o r o f o r m removed w i t h the a i d o f a r o t a r y e v a p o r a t o r .
The r e s u l t i n g c l e a r y e l l o w o i l s h o u l d be n e a r l y pure
X as the f r e e b a s e .
The o i l y p r o d u c t may c o n t a i n
u n r e a c t e d , - d i e t h y l a m i n o e t h a n o l , which was r e a d i l y
d e t e c t e d by NMR. I f the a m i n o a l c o h o l was p r e s e n t
the o i l y m a t e r i a l was washed by the f o l l o w i n g
procedure :
To the p r o d u c t , i n a l a r g e s e p a r a t o r y f u n n e l ,
was added 250 ml c o l d ( ^ 4 ) w a t e r .
The m i x t u r e was
shaken g e n t l y and the l a y e r s a l l o w e d t o s e p a r a t e .
V i g o r o u s s h a k i n g r e s u l t e d i n a e m u l s i o n which s e p a
r a t e d very s l o w l y .
The o i l y l a y e r was then s e p a r a t e d
and added t o a f l a s k c o n t a i n i n g a p p r o x i m a t e l y 200 ml
c h l o r o f o r m o v e r anhydrous sodium s u l f a t e .
After
d r y i n g , the s o l u t i o n was f i l t e r e d and the c h l o r o f o r m
removed on the r o t a r y e v a p o r a t o r .
The r e s u l t i n g o i l
was X.
The r e a c t i o n sequence f o r s t e p I I I i s o u t l i n e d
on the f o l l o w i n g page and u s u a l l y r e s u l t e d i n 95-100%
o f the t h e o r e t i c a l y i e l d :
Step I V .
X, a p p r o x i m a t e l y 65 g , was d i s s o l v e d
i n 170 ml g l a c i a l a c e t i c a c i d .
The s o l u t i o n was
s t i r r e d i n an i c e b a t h u n t i l a c l e a r s o l u t i o n r e
sulted.
W h i l e v i g o r o u s l y s t i r r i n g the a c e t i c s o l u
t i o n , 17.8 g (0.22 moles) p o t a s s i u m i s o c y a n a t e was
g r a d u a l l y added.
The s o l u t i o n was a l l o w e d t o s t i r
f o r 15 minutes i n the i c e b a t h and t h e n an a d d i t i o n a l
t h r e e hours a t room t e m p e r a t u r e .
The p r o d u c t $ - N ' , N - d i e t h y l a m i n o e t h y l - 2 , 2 d i p h e n y l h y d a n t o a t e (XI) was i s o l a t e d as the HSO4
s a l t by the f o l l o w i n g p r o c e d u r e :
A f t e r t h r e e hours s t i r r i n g a t room t e m p e r a t u r e ,
125 ml methanol was added to the r e a c t i o n m i x t u r e .
To a s e p a r a t e 125 ml p o r t i o n o f m e t h a n o l , 16.7 ml
(30 g , 0.3 moles) c o n c e n t r a t e d (98%)
sulfuric
,

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

STELLA

E T

AL.

Chemistry of a 5,5-Diphenylhydantoin Pro-drug

C= 0

HOC H -N

(IX)

HCI

M
/
C-OC H -N
2

NH 2

CO
2

(X)
a c i d was c a r e f u l l y added.
A f t e r the s u l f u r i c
a c i d / m e t h a n o l s o l u t i o n had c o o l e d t o room temperat u r e , i t was added t o the methanol r e a c t i o n m i x t u r e
s o l u t i o n with vigorous s t i r r i n g .
The HSO4 s a l t o f
p r o d u c t , X I , was t h e n p r e c i p i t a t e d by the a d d i t i o n
of e t h e r .
(Ether was added u n t i l no f u r t h e r p r e c i p i t a t i o n occurred.
Up t o 2 l i t e r s o f e t h e r may be
required.)
The r e a c t i o n sequence i s o u t l i n e d on the f o l lowing page:_
The HSO4 s a l t (XI) was c o n v e r t e d t o the S 0
salt
(XII) by the f o l l o w i n g p r o c e d u r e :
P r o d u c t X I , 47 g (0.1 m o l e ) , was d i s s o l v e d i n
1500 ml water (a c l e a r s o l u t i o n may n o t r e s u l t i f
p r o d u c t , X I , was i m p u r e ) .
Ammonium s u l f a t e , 1425 g ,
was d i s s o l v e d i n 2000 ml water and the s o l u t i o n
filtered.
The s o l u t i o n o f XI was t h e n f i l t e r e d
i n t o the same f l a s k and the m i x t u r e s t i r r e d .
After
c r y s t a l l i z a t i o n had commenced, the f l a s k was c o o l e d
t o 0 - 5 and l e f t f o r 3-4 hours a t t h i s temperature
or stored overnight i n a r e f r i g e r a t o r .
The c r y s t a l s were r e c o v e r e d by f i l t r a t i o n .
After
the c r y s t a l s had been d r i e d on the f i l t e r , the
c r y s t a l s were added t o 10 ml o f the w a t e r .
The m i x t u r e was s t i r r e d t h o r o u g h l y and the w a t e r removed by
vacuum f i l t r a t i o n .
=

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

163

164

PRO-DRUGS

1)KCN0/CH C00H
3

O C

2 4~
H

N <

(X)

HSO

(XI)
A f t e r a l l apparent t r a c e s o f m o i s t u r e had been
removed, the c r y s t a l s were t r a n s f e r r e d t o a f l a s k
c o n t a i n i n g 750 ml o f 95% e t h a n o l and s t i r r e d t h o r
oughly.
A t t h i s s t a g e , a c l e a r s o l u t i o n may n o t
r e s u l t due t o the p r e s e n c e o f (NH4)2S04.
The s o l u t i o n
was f i l t e r e d and the p r o d u c t X I I was p r e c i p i t a t e d by
the a d d i t i o n o f e t h e r ( a p p r o x i m a t e l y 2 l i t e r s was
required).
The e t h a n o l / e t h e r s o l u t i o n was s t i r r e d
f o r s e v e r a l hours o r o v e r n i g h t a t room temperature
and the c r y s t a l l i n e p r o d u c t r e c o v e r e d by f i l t r a t i o n .
The p e r c e n t a g e y i e l d i n the f i n a l c o n v e r s i o n t o
p r o d u c t XII was u s u a l l y q u i t e low (30-50%).
I t should
be p o i n t e d o u t t h a t t h i s s t e p c o u l d be improved by
a r e c y c l i n g o f the aqueous s o l u t i o n .
Compound, X I I , o r the h e m i - s u l f a t e s a l t o f - ' ,
N - d i e t h y l a m i n o e t h y l - 2 , 2 - d i p h e n y l h y d a n t o a t e w i l l be
r e f e r r e d t o as Pro-DPH f o r c o n v e n i e n c e ; t h i s was the
d e r i v a t i v e used i n a l l subsequent s t a b i l i t y and
animal s t u d i e s .
T a b l e I g i v e s the m e l t i n g p o i n t s , m o l e c u l a r
w e i g h t s , % e q u i v a l e n t o f DPH and s o l u b i l i t y o f a
number o f - , N - d i e t h y l a m i n o e t h y l - 2 , 2 - d i p h e n y l hydantoate s a l t s at 2 5 i n water.
The s u p e r i o r
aqueous s o l u b i l i t y o f t h e h e m i - s u l f a t e s a l t made i t
the c a n d i d a t e o f c h o i c e f o r i n i t i a l s c r e e n i n g
for anticonvulsant a c t i v i t y .
N o t e , t h a t DPH has
a s o l u b i l i t y o f a p p r o x i m a t e l y 2 mg/100 ml
,

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

STELLA

Chemistry of a 5,5-Diphenylhydantoin Pro-drug

E T A L .

H2
(XII)
or 0.02 mg/ml i n aqueous s o l u t i o n at pH pK at
25 which means that the increase i n aqueous s o l u
b i l i t y of Pro-DPH over DPH i s about a f a c t o r of
15,000 or 9,000 i n terms of DPH equivalents.
a

Table I
Some p h y s i c a l properties of various s a l t forms of
-', -diethylaminoethyl-2,2-diphenylhydantoate.
1

Mol.
Wt.

%Equivalence Melting
Pt.
of Diphenylhydantoin

Solu- *
bility
at 25 C

HN0

432

58.3

173(dec)

22 mg/ml

HCL

405.5

62.1

188

23 mg/ml

49.7

138

8 mg/ml

145

301 mg/ml

Salt

S a l i c y l a t e 507
Sulfate

837.01

60.28

S o l u b i l i t y r e f e r s to the s o l u b i l i t y of the s a l t
i t s e l f and not DPH equivalence.
** This gives an aqueous s o l u t i o n of pH ^ 3.3.

The conversion of Pro-DPH to DPH was quantita


t i v e as checked by s p e c t r a l comparisons and t h i n
layer chromatography. The rate of c y c l i z a t i o n of
Pro-DPH to DPH was followed at 25 and 37 to simulate
storage conditions and p h y s i o l o g i c a l conditions. A l l
k i n e t i c measurements were e i t h e r c a r r i e d out d i r e c t l y
i n the thermostated c e l l compartment of a recording
spectrophotometer, Cary Model 14, or i n ampoules
placed i n a constant temperature water bath. The

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

165

166

PRO-DRUGS

formation of DPH was followed spectrophotometrically


at 245 nm with the rate constants determined from
p l o t s of log (Aoo -A ) versus time, where Aoo and A
were the absorbances at i n f i n i t y and time t respec
tively.
As an a l t e r n a t i v e , Pro-DPH and/or DPH were
followed by high pressure l i q u i d chromatography a f t e r
extraction from the aqueous buffer s o l u t i o n s . A l l
reactions were c a r r i e d out i n aqueous buffered solu
tions of constant i o n i c strength. However, no a t
tempts to extrapolate to zero b u f f e r concentration
were made.
The observed rate constants and h a l f - l i v e s f o r
the conversion of Pro-DPH at 25 and 37 i n aqueous
buffered solutions t
Figure 1 shows a p l o
observed rate of c y c l i z a t i o n of Pro-DPH to DPH,
versus pH. Also shown i n t h i s figure i s the data
for the c y c l i z a t i o n of the equivalent pro-drug de
r i v a t i v e of mephenytoin and 3-N ,'-diethylaminoethyl-5-methyl-2,2-ethylphenylhydantoate the equiva
lent pro-drug d e r i v a t i v e of 3-methyl-5,5-ehtylphenylhydantoin. Note that the conversion of Pro-DPH to
DPH at 37 i s very e f f i c i e n t at pH 7.4, the h a l f l i f e i s 6.8 minutes, but at pH 3 the h a l f - l i f e i s
about 130 hours. At 25 and pH 3 the h a l f - l i f e f o r
the conversion of Pro-DPH to DPH was found to be
approximately 450 hours and at 4, r e f r i g e r a t o r
temperature, the h a l f - l i f e was 4000 hours.
I t should be noted that i n terms of s t a b i l i t y the
determining factor i n a possible reconstituted lyophil i z e d dosage form of Pro-DPH w i l l not be the loss of
Pro-DPH per se but more probably the gradual formation
of DPH and i t s subsequent p r e c i p i t a t i o n as the satura
t i o n s o l u b i l i t y of DPH i s reached. The formation of a
saturated s o l u t i o n of DPH w i l l be a function of the
i n i t i a l concentration of the Pro-DPH. Since one
mole of Pro-DPH released two moles of DPH
t

fc

[DPHft

= 2k [Pro-DPHl

(eq. 1)

^ t
o
/ci [DPHN
where V d s
is the i n i t i a l rate of formation of DPH
from Pro-DPH, [Pro-DPH] i s the i n i t i a l concentration
of Pro-DPH and k i s the pseudo f i r s t order observed
rate constant f o r the conversion of Pro-DPH to DPH
under the designated experimental conditions. I f
d[DPHf\ i s expressed as mg/ml/hr and [Pro-DPH]
d

(.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

STELLA ET

AL.

Chemistry of a 5,5-Diphenylhydantoin Pro-drug

CO

-H

>

I
m

r-i
rd

--

rH

Cfl

0
^!

m
m

<d

*"

CM

4J
\

(d

4J
(d

<

>

En

CO

LO

C0

00

CO

C0

CM

00

rd

-
Cl

CO

m
CM

CM CO
I
i

m
CM

-
fd

CM

Q
0

-M

ffi

CM

*
CO

M
P4

CM

CO
I

C0

*>

CM

as

00

CM

00

Q
1
0

M-l

00

CM

00

CO

rH

CM

>
0

**

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

167

168

PRO-DRUGS

i s expressed i n mg/ml then


,d[DPH]. _ 2k[Molecular Weight DPH]fPro-DPH]
dt 'o
[Molecular Weight Pro-DPH]
^

= k [Pro-DPH]
1

2 )

(eq. 3)

where k = 154 10~ 252 2/837 = 9.27 "


hours"! t 25 and at a pH of 3. Table I I I gives
the calculated zero order i n i t i a l rates of formation
of DPH from Pro-DPH as a function of i n i t i a l Pro-DPH
concentration at 25 and pH 3 and the time f o r
DPH to p o t e n t i a l l y begin nucleating from s o l u t i o n
(t
).
The s o l u b i l i t
i s assumed to be 2
that no DPH was present i n the i n i t i a l sample of
Pro-DPH so the time before p r e c i p i t a t i o n , i . e . ,
t p p t e , i s an o p t i m i s t i c estimate. The value of t t
may vary depending on the occurrence of super satura
ted solutions although t h i s a p r i o r i does not seem
l i k e l y i n l i g h t of the strong c r y s t a l l a t t i c e
a

p p t e

p p

Table I I I
Apparent zero order i n i t i a l rate of formation of
DPH from Pro-DPH as a function of i n i t i a l Pro-DPH
concentration at 25 and pH 3 and the p o t e n t i a l
t
based on Equations 1-3 and a s o l u b i l i t y of
Dt>! at 25 and pH 3 of 2 mg/100 ml.
t

t .
(min)
PPte

[Pro-DPH]

[d[DPH]/dt)

25 mg/ml

2.31 10~ mg/ml/hour

50 mg/ml

52 min

4.63 10"" mg/ml/hour

26

100 mg/ml

9.26 10" mg/ml/hour

13

200 mg/ml

1.85 lO^mg/ml/hour

of the formed DPH. Figure 2 i l l u s t r a t e s the t h e o r e t i


cal i n i t i a l rate of formation of DPH from Pro-DPH.
The time required f o r DPH p r e c i p i t a t i o n to occur from
solutions of Pro-DPH was determined experimentally
and compared to these predicted figures,
The t t
did not appear to c o r r e l a t e w e l l with t t e pre
dicted from the known rates of c y c l i z a t i o n of Pro-DPH
to DPH. Table IV gives some of the experimentally
determined t u r b i d i t y times f o r various i n i t i a l con
centrations of Pro-DPH.
p p

p p

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

STELLA

ET

AL.

Chemistry of a 5,5-Diphenyhydantoin Pro-drug

-51
0

I
1

I
2

I
3

I
4

I
5

I
6

1
7

i
8

1
9

L
10

PH

Figure 1. Phts of log k vs. pH for the


conversion of Pro-DPH to DPH at 37 ()
and at 25 (). Also included are data for
conversion of IV to I at 37 () and the
conversion of -', N'-diethylaminoethyl5-methyl-2,2-ethylphenylhydantoate to 3methyl-2,2-ethylphenylhydantoate at 37 ()
in aqueous buffered solutions.
ob8d

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

169

170

PRO-DRUGS

Table IV
Experimental Determination of T u r b i d i t y Time as a
Function of I n i t i a l Pro-DPH Concentration.
Concentration of Pro-DPH i n
mg/ml i n d i s t i l l e d water

pH

Time f o r T u r b i d i t y
Development

83.25

3.6

36 Minutes

83.25

3.7

15 Minutes

1 Hour 10 Minutes

208.1

2 Hours 10 Minutes

246.75

3.5

36 Minutes

124.84
166.5

To overcome the problem of DPH p r e c i p i t a t i o n , the


following approaches were attempted. The f i r s t approach used a solvent i n which DPH was more soluble.
This allowed more DPH to form before i t began to prec i p i t a t e out. The s o l u b i l i t y of DPH i n the current
DPH i n j e c t i o n vehicle of 40% propylene g l y c o l , 10%
alcohol and 50% water was 1.07 mg/ml. This meant
that a s o l u t i o n of Pro-DPH of 100 mg/ml and c y c l i z i n g
to DPH with a h a l f - l i f e of 450 hours a t 25 would
remain c l e a r f o r 46 hours. This approach would suff e r from the problem that some of the t o x i c i t y associated with the current DPH i n j e c t i o n may i n part
be due to the presence of propylene g l y c o l (36_* 37J .
The second approach that could be used (although i t
would not be s u i t a b l e f o r an I.V. i n j e c t i o n dosage
form) would require a sparingly water soluble but
r e a d i l y d i s s o c i a t a b l e s a l t of Pro-DPH. A sparingly
soluble s a l t (perhaps i n the presence of an excess of
the counter anion) could be used to e f f e c t a suspension dosage form. On I.M. i n j e c t i o n , the absorption
of the common ion into general c i r c u l a t i o n and subsequent d i s s o c i a t i o n of the s l i g h t l y soluble s a l t should
r e s u l t i n reasonable blood l e v e l s of DPH. This technique was attempted using the s a l i c y l a t e s a l t of XI
(Pro-DPH s a l i c y l a t e ) as a model. Pro-DPH s a l i c y l a t e
was found to have an aqueous s o l u b i l i t y i n water of
8 mg/ml. I f 100 mg of Pro-DPH s a l i c y l a t e was suspended i n one ml of water the amount of dissolved
Pro-DPH s a l i c y l a t e would be 8 mg, i . e . , saturation

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

STELLA

E T

Chemistry of a 5,5-Diphenylhydantoin Pro-drug

AL.

solubility.
The c y c l i z a t i o n o f Pro-DPH t o DPH a t
2 5 and pH 4.5 (the pH n e c e s s a r y t o m i n i m i z e d i s s o c i a
t i o n o f Pro-DPH s a l i c y l a t e ) has a h a l f - l i f e o f 180
hours o r k k
i s 3.85 10"3 h o u r s " .
The i n i t i a l
r a t e o f f o r m a t i o n o f DPH o r ( d [ D P H ] / d t ) from t h i s
s a t u r a t e d s o l u t i o n would be 0.015 mg/ml/hour, i . e . ,
i t would be 1.3 hours b e f o r e DPH would b e g i n
p r e c i p i t a t i n g out.
T a b l e V shows the e f f e c t
o f added s a l i c y l a t e a n i o n on the s o l u b i l i t y o f P r o DPH s a l i c y l a t e .
In the p r e s e n c e o f 0.1 M sodium
s a l i c y l a t e , the s o l u b i l i t y i s lowered t o 1 mg/ml so
t h a t the i n i t i a l r a t e o f f o r m a t i o n o f DPH becomes
0.0010 mg/ml/hour and i t would be 10.5 hours b e f o r e
DPH would b e g i n t o p r e c i p i t a t e .
The c a l c u l a t e d
s o l u b i l i t y product o
d a t a was found t o be 3.04 1 0 ~ M .
0

S c i

Table V
Aqueous S o l u b i l i t y o f Pro-DPH S a l i c y l a t e as
F u n c t i o n o f Added S a l i c y l a t e A n i o n
Pro-DPH s a l i c y l a t e
s o l u b i l i t y i n mg/ml

6.8

0.01

2.9

0.05

1.5

0.10

S i n c e the pK

s o l u b i l i t y product w i l l
o f about two

(^ 1.5

SP

3.04xlO" M
4

of s a l i c y l i c i s ^ 3.0,

as the pH i n c r e a s e s ,

Calculate

Cone, o f
added Sodium S a l i
cylate

8.7

or

the

n o t change s i g n i f i c a n t l y

but w i l l
1"

d e c r e a s e by a

factor

M ) a t pH 3 . 0 .
2

Discussion
The s y n t h e s i s o f Pro-DPH was n o t s h o r t b u t a t
the same time d i d n o t r e q u i r e any h i g h degree o f
sophistication.
On a commercial b a s i s , the c o n v e r
s i o n o f V I , 2 , 2 - d i p h e n y l g l y c i n e , t o IX might be
e f f e c t e d by a one s t e p r e a c t i o n w i t h phosgene r a t h e r
than the two s t e p approach t a k e n .
O b v i o u s l y the c o s t
o f p r o d u c i n g the Pro-DPH would be h i g h e r than DPH

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

171

172

PRO-DRUGS

i t s e l f s i n c e DPH i s the p r i m a r y s t a r t i n g m a t e r i a l and


the s y n t h e s i s o f the Pro-DPH i n v o l v e s a t l e a s t a f i v e s t e p pathway.
The v a r i o u s s a l t forms o f Pro-DPH and t h e i r
v a r i a b l e aqueous s o l u b i l i t y ( r e f e r t o T a b l e I) r a i s e s
an i n t e r e s t i n g and o f t e n o v e r l o o k e d p o i n t .
The
o r i g i n a l o b j e c t i v e o f t h i s p r o p o s a l was t o s y n t h e s i z e
a water s o l u b l e b i o r e v e r s i b l e p r o - d r u g o f DPH which
c o u l d be a d m i n i s t e r e d o r a l l y , i n t r a m u s c u l a r l y o r
intravenously i n a p h y s i o l o g i c a l l y compatible v e h i c l e .
The p r o - d r u g s h o u l d not p r e c i p i t a t e on i n j e c t i o n i n
the body and s h o u l d q u a n t i t a t i v e l y r e v e r t t o DPH.
The s o l u b i l i t y o f h y d r o c h l o r i d e s a l t s o f amines f o r
o r a l d e l i v e r y i s s u p p r e s s e d by the common i o n e f f e c t
o f endogenous h y d r o c h l o r i d i
A l s o i t i s not uncommo
o f l a r g e h y d r o p h o b i c amines t o be among the l e a s t
water s o l u b l e s a l t .
As n o t e d i n r e c e n t d i s c l o s u r e s
and i n new drugs approved f o r c l i n i c a l use, the use
o f s a l t s o t h e r than h y d r o c h l o r i d e s i s a welcome ass u r a n c e t h a t the importance o f the c o u n t e r a n i o n on
the aqueous s o l u b i l i t y o f h y d r o p h o b i c amines s a l t s i s
being recognized.
The c h o i c e o f a good c o u n t e r a n i o n
f o r an amine drug i s o f t e n d i f f i c u l t t o p r e d i c t a
p r i o r i i n t h a t the d e t e r m i n i n g f a c t o r i n the aqueous
s o l u b i l i t y o f amine s a l t s i s o f t e n the s t r e n g t h o f the
formed c r y s t a l l a t t i c e i n the s o l i d phase r e l a t i v e t o
the s o l v a t i o n energy r e l e a s e d on s o l u t i o n . A l t h o u g h
the s o l v a t i o n energy a s p e c t s o f s o l u b i l i t y a r e o f t e n
w e l l u n d e r s t o o d , s o l i d s t a t e geometry and s t a c k i n g
o f the c r y s t a l l a t t i c e can o n l y be d e t e r m i n e d by
X-ray t o t a l s t r u c t u r e d e t e r m i n a t i o n which i s o f l i t t l e
v a l u e t o the p r a g m a t i s t l o o k i n g f o r a q u i c k answer.
N e v e r t h e l e s s , T a b l e I does show t h a t o v e r an o r d e r o f
magnitude i n c r e a s e i n aqueous s o l u b i l i t y c o u l d be
e f f e c t e d by the j u d i c i o u s s c r e e n i n g o f more than one
s a l t form o f a s u s p e c t e d p h a r m a c o l o g i c a l l y a c t i v e
agent whose aqueous s o l u b i l i t y r e q u i r e d o p t i m i z a t i o n .
The p a r t i a l l o g kobsd v e r s u s pH p r o f i l e f o r the
c o n v e r s i o n o f Pro-DPH t o DPH a t 25 and 37 shown i n
F i g u r e 1 r e v e a l s the pH o f maximum s t a b i l i t y t o be
^2.5.
The c o n v e r s i o n appears t o have an a c i d c a t a l y z e d and a s p e c i f i c base c a t a l y z e d component as
w e l l as a spontaneous o r water c a t a l y z e d component.
The i n f l e c t i o n c o r r e s p o n d i n g t o a d i f f e r e n c e i n
r e a c t i v i t y o f the s p e c i f i c base c a t a l y z e d r e a c t i o n
between the p r o t o n a t e d r e l a t i v e t o u n p r o t o n a t e d
form o f the Pro-DPH was not n o t e d because the h i g h e s t
pH s t u d i e d was 7.4.
As n o t e d i n our e a r l i e r s t u d y ,
a p a r t i a l p l a t e a u i n g s h o u l d have been n o t e d between

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

STELLA

Chemistry of a 5,5-Diphenylhydantoin Pro-drug 173

E TA L .

pH 8 and 9.5 (30.)


P a
group i n
Pro-DPH i s e x p e c t e d t o be ^ 8 . 5 - 9 . 0 a t 3 7 (30).
K i n e t i c a l l y the c o n v e r s i o n o f Pro-DPH t o DPH
may be d e f i n e d by e q u a t i o n 4.
T

obsd

[ H + ] a

-OH
r

k ' - ^ ] (1-a)

(eq. 4)

where k b s d i s the o b s e r v e d r a t e c o n s t a n t f o r t h e
c o n v e r s i o n o f Pro-DPH a t a g i v e n t e m p e r a t u r e , k +
i s an a c i d c a t a l y z e d c o n s t a n t , k i s a spontaneous
o r water c a t a l y z e d r a t e c o n s t a n t , k is a specific
base c a t a l y z e d r a t e c o n s t a n t r e p r e s e n t i n g t h e a t t a c k
o f the u r e i d o a n i o n
p r o t o n a t e d form o f Pro-DPH and k ' is a specific
base c a t a l y z e d r a t e c o n s t a n t r e p r e s e n t i n g t h e a t t a c k
o f t h e u r e i d o a n i o n a t t h e e s t e r l i n k a g e o f the u n p r o t o n a t e d form o f P r o - D P H .
A l t h o u g h w i t h Pro-DPH
no s o p h i s t i c a t e d p h y s i c a l o r g a n i c study was c a r r i e d
o u t ( t h a t was n o t t h e o b j e c t i v e o f t h i s study) by
i n f e r e n c e t o o u r e a r l i e r more d e t a i l e d k i n e t i c
s t u d y o f the c o n v e r s i o n o f IV t o I the l o g k
v e r s u s pH p r o f i l e and E q u a t i o n 4 were c o n s i s t e n t w i t h
Scheme I I .
A t p H ' s g r e a t e r t h a n 5 b u t l e s s t h a n the p K
o f t h e amino g r o u p , t h e r a t e d e t e r m i n i n g s t e p appears
t o be t h e i n t r a m o l e c u l a r a t t a c k o f t h e u r e i d o a n i o n
on the n e i g h b o r i n g e s t e r l i n k a g e .
T h i s was c o n s i d e r e d
t o be t h e most l i k e l y mechanism because o f t h e l a c k
o f b u f f e r c a t a l y s i s (30).
The spontaneous r a t e c o n
s t a n t k p r o b a b l y i n v o l v e s the a t t a c k o f t h e n e u t r a l
u r e i d o group a g a i n v i a an i n t r a m o l e c u l a r r e a c t i o n on
the n e i g h b o r i n g e s t e r l i n k a g e .
These r e a c t i o n s
o f c o u r s e i n v o l v e Pro-DPH c o n v e r t i n g t o DPH w i t h t h e
subsequent e x p u l s i o n o f t h e , - d i e t h y l a m i n o e t h a n o l
leaving group.
The minor n a t u r e o f t h e a p p a r e n t
s p e c i f i c a c i d c a t a l y z e d pathway which i n v o l v e s t h e
a t t a c k o f t h e u r e i d o group on the p r o t o n a t e d e s t e r
f u n c t i o n i s p r e d i c t a b l e i n t h a t p r o t o n a t i o n o f the
e s t e r f u n c t i o n would n o t be f a v o r a b l e due t o e l e c t r o
s t a t i c r e p u l s i o n . The d i m i n u t i v e n a t u r e o f t h i s
pathway i s c o n s i s t e n t w i t h e a r l i e r h y d r o l y s i s s t u d i e s
on o t h e r amino e s t e r s such as p r o c a i n e (.37) and
a t r o p i n e (38^,39) t where l i t t l e s p e c i f i c a c i d c a t a l y z e d
h y d r o l y s i s was s i m i l a r l y n o t e d .
These r e a c t i o n s
d i d n o t i n v o l v e an i n t r a m o l e c u l a r r e a c t i o n
b u t were normal i n t e r m o l e c u l a r h y d r o l y s i s r e a c t i o n s .
T a b l e I n o t e d t h e 15,000 f o l d i n c r e a s e i n aqueous
s o l u b i l i t y o f Pro-DPH o v e r DPH. I n terms o f DPH
Q

Q H

0 H

o t ) S d

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

H 0

R,

2 5 2

K,

( C

C-OC H -N(C H )

+H

O R

li i

- 2 4-7

NH-C-NH

\ /

-H

Scheme

II

HYDANTOIN

+H

+H

-H

K
2

-H

i - \

\ /

R_

NH-C-N

+H

O R

II l

NH-C-NH

C-OCJ.-N(C

3.

STELLA

ET

AL.

Chemistry of a 5,5-Diphenylhydantoin Pro-drug

equivalents t h i s was a factor of 9,000. As noted


e a r l i e r , the r e s u l t i n g formulation problem, i . e . ,
the rapid formation of a saturated s o l u t i o n of DPH
from a sample of Pro-DPH, presented another i n t e r e s t ing dimension to the problem. The experimentally determined t u r b i d i t y times or t p
as shown i n Table IV
as a function of i n i t i a l Pro-DPH concentration were
not consistent with the t h e o r e t i c a l values of Table
III.
Increasing i n i t i a l Pro-DPH concentration should
have led to a shorter t u r b i d i t y time. This anomalous
behavior could have r e s u l t e d from two possible
sources: a) The Pro-DPH formed a m i c e l l a r s o l u t i o n
which helped to p a r t i a l l y s o l u b i l i z e the formed DPH
thus lengthening tppte? ) The Pro-DPH formed m i c e l l e s
but, i n the m i c e l l a
slowly to DPH than i
of these postulated mechanisms were v e r i f i e d but the
f a c t that Pro-DPH i n the presence of plasma was apparently converted s l i g h t l y more slowly to DPH (see
paper by Glazko et al.) than i n the absence of plasma
under the same conditions of pH and temperature suggested that bound Pro-DPH may not be as r e a d i l y
converted to DPH as unbound Pro-DPH. This f i n d i n g
would tend to favor the second postulate as the
source of the anomalous t u r b i d i t y data.
p

t e

Conclusion
The o r i g i n a l objective of t h i s work was to u t i l i z e the pro-drug approach to produce a b i o - r e v e r s i b l e
DPH d e r i v a t i v e with good aqueous s o l u b i l i t y . I t was
hoped that the d e r i v a t i v e would have good i n vivo
DPH release c h a r a c t e r i s t i c s (see the paper of Glazko
et a l . following) and lead to a superior form of DPH.
A water soluble b i o r e v e r s i b l e pro-drug of DPH was
synthesized and i t s p h y s i c a l and chemical properties
studied. The pro-drug was an a c y c l i c form of DPH
which underwent an intramolecular reaction to regenerate the parent compound, DPH.
Under simulated
p h y s i o l o g i c a l conditions, the h a l f - l i f e f o r the conversion of the Pro-DPH to DPH was approximately seven
minutes i l l u s t r a t i n g that enzyme mediation i n the r e generation of the parent compound was unnecessary.
The r e l a t i v e s t a b i l i t y of Pro-DPH under a c i d i c s o l u t i o n s , pH ^ 3, allowed a l y o p h i l i z e d powder to be
formed. When reconstituted with water, the extremely
poor aqueous s o l u b i l i t y of the gradually formed DPH
did create problems by producing t u r b i d solutions rel a t i v e l y q u i c k l y . The e f f e c t of the counter anion

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

175

176

PRO-DRUGS

on the aqueous s o l u b i l i t y of the amine pro-drug was


shown to be an important factor i n producing a
s a t i s f a c t o r i l y water soluble pro-drug.
Other Attempts at Pro-Drug Forms of Hydantoins
In the course of the i n v e s t i g a t i o n on the poss i b l e usefulness of hydantoic acid esters as pro-drugs
of hydantoins, the acid catalyzed c y c l i z a t i o n of hydant o i c acids themselves, was investigated i n the pH range
0-2.
The reaction was found to follow Scheme I I I . Det a i l s of the k i n e t i c s and the e f f e c t s of the R , R and
R3 groups on the closure rate have been reported (40);
of s p e c i f i c i n t e r e s t were 2,2-diphenylhydantoic acid
and 2-ethyl-5-methyl-2-phenylhydantoi
t o i c acids of DPH an
respectively
objectiv
the study of the acid catalyzed c y c l i z a t i o n of the hydantoic acids was to observe i f when taken o r a l l y , the
closure of the hydantoic acids to t h e i r respective hydantoin was s u f f i c i e n t l y fast to allow hydantoic acid
to be converted to the hydantoin under p h y s i o l o g i c a l pH
conditions. I t was found that only i n the acid pH r e gion was the closure rate observable. The study showed
that i n the pH range 0 to 2 both of the respective hydantoic acids of DPH and I were q u a n t i t a t i v e l y converted to the hydantoin.
The pH of stomach contents i n a fasted human i s
considered to be i n the range of 1 to 3 and the stomach
emptying time f o r small volumes of l i q u i d s i s v a r i a b l e .
If the hydantoic acids were to act as pro-drugs of hydantoins, they should c y c l i z e with h a l f - l i v e s at pH 1.5
and 37 of less than 15 minutes. At pH 1.5 and 50,
the h a l f - l i f e f o r the conversion of 2,2-diphenylhydant o i c acid to DPH was 12 hours while the h a l f - l i f e f o r
the conversion of 2-ethyl-5-methyl-2-phenyl hydantoic
acid to I was 40 minutes. I f a three f o l d decrease i n
rate i s assumed f o r a temperature drop of 50 to 37,
the h a l f - l i v e s f o r the conversions were w e l l outside
the range where the hydantoic acids might be considered
as s u i t a b l e pro-drugs of hydantoins.
Reference to l , 3 - d i a c y l - , 3 - a c y l - , and 1-acyl-, der i v a t i v e s of hydantoins have appeared i n t e r m i t t e n t l y i n
the l i t e r a t u r e . As discussed e a r l i e r , poor aqueous and
l i p i d s o l u b i l i t y of hydantoins not substituted at the 1
and 3 positions i s due i n part to strong intermolecular
hydrogen bonding i n the c r y s t a l l a t t i c e . Acylation at
the number 3 p o s i t i o n , which displaces the a c i d i c proton, should lead to a reduction i n the intermolecular
hydrogen bonding i n the c r y s t a l l a t t i c e . This should
r e s u l t i n the increased l i p o p h i l i c i t y of hydantoins as
1

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

STELLA

E T

Chemistry of a 5,5-Diphenylhydantoin Pro-drug

A L .

R
2

I II

R-C-C-0

+H

NHCNH3
Il

K'
a

(A )
R

I II

CCOH

2

R CCOH

.+
+H

NHCNH
O

NHCNH
O R

(HA)
\

(H A )
2

,
product

I
C

/ '2
k

C=0
1

HN

NR

II
o

(H)
Scheme

III

where R , R , R = a l k y l ,
hydrogen s u b s t i t u e n t s .
1

aryl,

and/or

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

177

178

PRO-DRUGS

w e l l as a p o s s i b l e i n c r e a s e i n water s o l u b i l i t y .
E n g l i s h e t a l . ( 1 ) t e s t e d b o t h t h e i n v i t r o and
in vivo a c t i v i t y of various 3 - a l k y l , 3-acyl,
3-acyloxym e t h y l , and 3 - a l k o x y c a r b o n y l d e r i v a t i v e s (see s t r u c t u r e
below) o f t h e h y d a n t o i n , n i t r o f u r a n t o i n (II) a g a i n s t
v u l g a r i s .
They n o t e d l i t t l e c o r r e l a t i o n between i n

(ID
where R = - a l k y l ;
- C R where R i s an
a l k y l , a r y l o r a l k o x y g r o u p ; and - O ^ O R
where R
i s an a l k y l o r a c y l g r o u p .
1

1 1

v i t r o and i n v i v o a c t i v i t y f o r a number o f compounds.


Compounds w i t h i n v i v o a c t i v i t y s i m i l a r t o I I i t s e l f
were 3 - h y d r o x y m e t h y l - and 3 - a c e t o x y m e t h y l n i t r o f u r a n toin.
A l l o f t h e 3 - a c y l d e r i v a t i v e s showed good a c t i v i t y w i t h t h e g r e a t e s t a c t i v i t y demonstrated by t h e C2
t h r o u g h C5 compounds.
In t h e 3 - a l k o x y c a r b o n y l d e r i v a t i v e s good a c t i v i t y was n o t e d f o r C^ t h r o u g h C4 d e r i v a tives.
The a u t h o r s n o t e d t h a t " t h e r o l e o f 3 - s u b s t i t u e n t s i n a l t e r i n g t h e c h e m i c a l and p h y s i c a l p r o p e r t i e s
i n f l u e n c i n g a b s o r p t i o n , e x c r e t i o n and t r a n s p o r t , e t c .
is recognized".
The f a c t t h a t a l l d e r i v a t i v e s showed
the e v e n t u a l appearance o f n i t r o f u r a n t o i n i t s e l f s u g g e s t s t h a t t h e a c t i v e m e d i c i n a l agent was n i t r o f u r a n t o i n a l t h o u g h t h i s was n o t a c t u a l l y v e r i f i e d i n t h e
study.
R e f e r e n c e t o 1 , 3 - d i a c y l , 3 - a c y l , and 1 - a c y l d e r i v a t i v e s o f DPH and 1 , 3 - d i a l k o x y c a r b o n y l ,
3-alkoxycarb o n y l , and 1 - a l k o x y c a r b o n y l d e r i v a t i v e s o f DPH have
been n o t e d ( 4 2 - 4 7 ) .
A number o f t h e s e d e r i v a t i v e s s y n t h e s i z e d by Umemoto (42-43) were s u b j e c t e d t o a n i m a l
t r i a l s by Nakamura e t a l . (26-29) and one d e r i v a t i v e ,
3 - c a r b e t h o x y - 5 , 5 - d i p h e n y l h y d a n t o i n (III) has a l s o been
s u c c e s s f u l l y s u b j e c t e d t o human t e s t i n g ( 4 8 - 4 9 ) . A n o t h e r d e r i v a t i v e 3 - a c e t y l - 5 , 5 - d i p h e n y l h y d a n t o m (XIV)
a l s o showed e a r l y promise b u t d i d n o t appear t o be as
e f f e c t i v e as I I I .
XIV had a mp o f 1 3 3 - 1 3 5 w h i l e I I I

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3.

STELLA E T A L .

Chemistry of a 5,5-Diphenylhydantoin Pro-drug 179

C= 0
HN

N C - 0 C H
'
Il
o

(III)
had a mp o f 138-140, i . e . , a c y l a t i o n a t t h e 3 p o s i t i o n
o f DPH caused a d e c r e a s e i n mp o f a p p r o x i m a t e l y 1 6 0
when compared t o DPH. T h i s o c c u r r e d as t h e a c y l group
d i s p l a c e d t h e most a c i d i c p r o t o n o f DPH [ p K 8 . 3 ,
(1/9.)] r e s u l t i n g i n d e c r e a s e d i n t e r m o l e c u l a r hydrogen
bonding i n the c r y s t a l l a t t i c e .
S e v e r a l s t u d i e s have
s u g g e s t e d t h a t 1 - a c y l - and l , 3 - d i a c y l - 5 , 5 - d i p h e n y l h y d a n t o i n s had l i t t l e a n t i c o n v u l s a n t a c t i v i t y and were
e s s e n t i a l l y e x c r e t e d as 1 - a c y l - 5 , 5 - d i p h e n y l h y d a n t o i n
whereas 3 - a c y l d e r i v a t i v e s had c o n s i d e r a b l e a n t i c o n v u l s a n t a c t i v i t y and i n t h e c a s e o f I I I t h e a c t i v i t y a p p e a r e d t o be g r e a t e r than DPH i t s e l f .
These r e s u l t s
suggest t h a t 3 - a c y l d e r i v a t i v e s are converted to the
p a r e n t h y d a n t o i n whereas t h e 1 - a c y l d e r i v a t i v e s a r e r e s i s t a n t to b i o t r a n s f o r m a t i o n to the parent hydantoin.
Nakamura e t a l . (26-29) have shown t h a t I I I demonstrat e d h i g h e r b l o o d l e v e l s and h i g h e r CNS l e v e l s o f DPH i n
r a t s and dogs when compared t o DPH i t s e l f .
Both drugs
were g i v e n as o r a l s u s p e n s i o n s .
Two human t r i a l s comparing I I I t o DPH by K i s h i e t
a l . (48) and Taen e t a l . ( 9 ) were e n c o u r a g i n g e s p e c i a l l y i n t h e i r conclusions of diminished side e f f e c t s
o f I I I r e l a t i v e t o DPH.
We have r e c e n t l y s y n t h e s i z e d a s e r i e s o f 3 - a l k o x y c a r b o n y l - 5 , 5 - d i p h e n y l h y d a n t o i n d e r i v a t i v e s w i t h a view
o f u t i l i z i n g t h e lower m e l t i n g p o i n t s and h i g h e r l i p i d
s o l u b i l i t i e s o f the d e r i v a t i v e s t o e f f e c t a s o f t g e l a t i n c a p s u l e dosage form o f DPH.
3-Hexoxycarbonyl-5,5d i p h e n y l h y d a n t o i n (XV) has a m e l t i n g p o i n t o f 8 6 and
was found t o be s o l u b l e and s t a b l e i n sesame o i l , p e a nut o i l , e t c .
XV was a l s o s t a b l e i n t h e b l a n d o i l s i n
the p r e s e n c e o f s u r f a c t a n t s such as Tween 80.
The b i o a

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

180

PRO-DRUGS

6 13

(XV)
a v a i l a b i l i t y o f DPH
form o f XV w i l l be r e p o r t e d a t a l a t e r d a t e .
The h y d a n t o i n , 5 - e t h y l - 5 - p h e n y l h y d a n t o i n (XVI) o r
N i r v a n o l was marketed as an a n t i c o n v u l s a n t drug b u t
f e l l i n t o d i s f a v o r and was removed from the market b e
cause o f h i g h t o x i c i t y .
I i s d e m e t h y l a t e d i n man and
dog t o XVI and i t seems l i k e l y t h a t the a n i t c o n v u l s a n t
a c t i v i t y o f I (see Scheme IV) i s , i n p a r t , due t o i t s
d e m e t h y l a t e d p r o d u c t , XVI (5JD) .
T h e r e f o r e , i t would
seem t h a t I i t s e l f i s a p r o - d r u g o f X V I .
P r o - d r u g s o f h y d a n t o i n s are not numerous.
This
s e c t i o n has r e v i e w e d what d e r i v a t i v e s have been attemp
t e d w i t h the hope t h a t i t might s t i m u l a t e f u r t h e r i n
v e s t i g a t i o n i n t o more e f f i c i e n t l y absorbed h y d a n t o i n
pro-drugs.

C= 0
N - CH

HN

oxidation

in vivo

C=0

C-

NH

HN
II

II

(XVI)

(I)

Scheme IV

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3. STELLA ET AL.

Chemistry of a 5,5-Oiphenyhydantoin Pro-dru

Literature Cited
1. Arnold, ., Gerber, ., and Levy, G., Can. J.
Pharm. Sci. (1970), 5, 89.
2. Martin, C. ., Rubin, M., O'Malley, W., Garaguai,
V. F., and McCauley, C. ., Pharmacologist (1968),
10, 167.
3. Rail, L., Med. J. Aust. (1968), ii, 339.
4. Suzuki, T., Saitoh, ., and Hishihara, ., Chem.
Pharm. Bull. (1970), 18, 405
5. Blum, M. R., Riegelman, S., and McGilveray, I.,
Paper 12 presented to the Basic Pharmaceutics
Section, APhA Academy of Pharmaceutical Sciences
San Francisco,
6. Blum, M. R., Riegelman, ,
,
, Pape
12 presented to the Basic Pharmaceutics Section,
APhA Academy of Pharmaceutical Sciences, San
Francisco, 1971.
7. Albert, ., Nature (1958), 182, 421.
8. Albert, ., "Selective Toxicity", 2nd. Ed., p.
30, Wiley, New York, N.Y., 1960.
9. Agarwal, S. P., and Blake, M. I., J. Pharm. Sci.
(1968), 57, 1434.
10. Glazko, A. J., Pharmacology (1972), 8, 163.
11. Sohar, P., Acta Chim. Acad. Sci. Hung. (1968),
57, 425.
12. "Dilantin", Formulary Monograph from Parke,
Davis and Co., Detroit, Michigan 48232.
13. Atkinson, Jr., A. J., and Davison, R., Ann. Rev.
Med. (1974), 25, 99.
14. Frank, J. T., Drug Intel. Clin. Pharm. (1973),
7, 287.
15. Sachtler, G., Drug Intel. Clin. Pharm. (1973).
7, 418.
16. Tobias, D. C., and Kellick, . ., Drug Intel.
Clin. Pharm. (1973), 7, 418.
17. Catania, P. ., Drug Intel. Clin. Pharm. (1973),
7, 418.
18. Chan, N. L., Drug Intel. Clin. Pharm. (1973),
7, 419.
19. Frank, J. T., Drug Intel. Clin. Pharm. (1973),
7, 419.
20. Baldwin, J., and Amerson, . ., Amer. J. Hosp.
Pharm. (1973), 30, 837.
21. Rowland, ., "Clinical Pharmacology", p.27,
Macmillan Co., New York, N.Y., 1972.
22. Serrano, E., Roye, D., Hammer, R., and Wilder,
B., Neurology (1973), 23, 311.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

182

PROD
- RUGS

23.

Wilensky, ., and Lowden, A., Neurology (1973),


23, 318.
Lagos, J., J. Amer. Med. Assoc. (1972), 220, 726.
Hopkins, I. J., and Rooney, J. C., Med. J. Aust.
(1969), 2, 278.
Nakamura, K., O'Hasi, K., Nakatsuji, K., Hiroka,
T., Fujimoto, ., and Ose, S., Arch. Int.
Pharmacodyn. (1965), 156, 261.
Nakamura, ., and Masuda, ., Arch. Int. Pharma
codyn. (1966), 164, 255.
Nakamura, ., Masuda, ., and Nakatsuji., Arch.
Int. Pharmacodyn. (1967), 165, 103.
Nakamura, ., Masuda, Y. Nakatsuji, ., and
Hiroka, T., Nauyn-Schmiedebergs Arch Pharmak
u. Exp. Path. (1966)
Stella, V., and Higuchi, T., J. Pharm. Sci.
(1973), 62, 962.
Hegarty, A. F., and Bruice, T. C., J. Amer. Chem.
Soc. (1970), 92, 6575.
Levine, R. R., Weinstock, J., Zirckle, C. S., and
McLean, R., J. Pharmacol. Exp. Ther. (1961), 131,
334.
Harper, N. J., J. Med. Pharm. Chem. (1959), 1,
467.
Harper, N. J., Progr. Drug Res. (1972), 4, 221.
Duschinsky, R., U. S. Patent 2,593,860 (1952),
to Hoffman-LaRoche, Inc., Nutley, N.J.
Louis, S., Kutt, ., and McDowell, F., Amer.
Heart J. (1967), 74, 523.
Higuchi, T., Havinga, ., and Busse, L. W., J.
Amer. Pharm. Assoc. Sci. Ed. (1950) ,39,405.
Kondritzer, . ., and Zvirblis, P., J. Amer.
Pharm. Assoc. Sci. Ed. (1957), 46, 531.
Zvirblis, P., Socholitsky, I., and Kondritzer,
. ., J. Amer. Pharm. Assoc. Sci. Ed. (1956),
45, 450.
Stella, V., and Higuchi, T., J. Org. Chem. (1973),
38, 1527.
English, A. R., McBride, T. J., Conover, L. H.,
and Gordon, P. ., Antimicrobial Agents Chemo
therapy (1966), 434.
Umemoto, S., J. Pharm. Soc. Jap. (1965), 84, 504.
Umemoto, S., J. Pharm. Soc. Jap. (1965), 84, 509.
Takamatsu, H., Umemoto, S., Fujimoto, ., and
Nakamura, K., U.S. patent 3,161,652, Dec. 15,
1964; Jap. App., Sept. 12, 1962 through C.A.
62, 7768d (1965).
Garcia Marquina, J. ., and Angles Besa, J. .,
Farm. Neuva. (1951), 16, 255.

24.
25.
26.
27.
28.
29.
30.
31.
32.
33.
34.
35.
36.
37.
38.
39.
40.
41.
42.
43.
44.
45.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

3. STELLA ET AL.

Chemistry of a 5,5-Diphenylhydantoin Pro

46. Angles Besa, J. M., Rev. Real Acad. Ciene Exact.


Fis. y Nat. Madrid (1949), 43, 193.
47. Garcia Marquina, J. M., andAnglesBesa, J. M.,
Farm Neuva. (1950), 15, 169.
48. Kishi, Y., Fukui, S., Maumara, S., and Aoki,
K., Brain and Nerve (1964), 16, 982.
49. Taen, S., Goto, Y., Sekiba, K., and Arai, S.,
Brain and Nerve (1964), 16, 977.
50. Butler, T. C., J. Pharmacol., (1952), 104, 299.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

4
The Metabolic Disposition of a Novel
5,5-Diphenylhydantoin Pro-drug
A. J. GLAZKO, W. A. DILL, R. H. WHEELOCK, R. M. YOUNG,
A. NEMANICH, and L. CROSKEY
Research Laboratories, Parke, Davis, and Co., Ann Arbor, Mich. 48106
V. STELLA and T. HIGUCHI
University of Kansas, Lawrence, Kans. 66044
5,5-Diphenylhydantoin (DPH; phenytoin; Dilantin )
has been used extensivel
oral and parenteral
its low water solubility (about 20 g/ml at 25 C),
parenteral formulations are dissolved in40%propylene
glycol and10%alcohol in water for injection, adjusted
to pH 12 to convert the acid to the sodium salt. A l
though this solution has been adequate for intravenous
use, there are clinical indications that intramuscular
use may result in slow absorption and low plasma levels
of DPH. One approach to this problem is based upon the
observation of Kozelka and Hine (1) that diphenylhydantoic acid undergoes ring closure to form DPH when heat
ed with strong acids. However, ring closure of diphenylhydantoic acid proceeds very slowly under physiolog
ical conditions.
Stella and Higuchi (2) studied a number of water
-soluble hydantoate esters and found that cyclization
occurred rapidly in neutral and alkaline solutions at
room temperature, apparently due to a specific base
-catalyzed intramolecular closure. As an outgrowth of
this work, Stella and Higuchi prepared the diethylami
noethanol ester of diphenylhydantoic acid, first as the
nitrate salt, and later as the hemisulfate (Pro-DPH).
These preparations were tested in the Parke-Davis Lab
oratories, and preliminary observations on metabolic
disposition were reported elsewhere (3). The structure
of this compound is shown in Figure 1.
*

Some Properties of Pro-DPH


The first preparation available for study was the
nitrate salt, which has a water solubility of 22 mg/ml
(representing the equivalent of 12.8 mg DPH per ml).
However, most of the experiments were carried out with
184
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

4.

GLAZKO

ET

AL.

Metabolic Disposition of 5,5-Diphenylhydantoin

the hemisulfate s a l t (Figure 1 ) , with a water s o l u b i l i


ty of about 308 mg/ml (representing the equivalent of
185 mg DPH per ml). This i s approximately 1 0 , 0 0 0 - f o l d
greater than the water s o l u b i l i t y of DPH. Both s a l t s
showed good s t a b i l i t y i n a c i d i c s o l u t i o n s , but were
converted r a p i d l y to DPH v i a r i n g closure i n a l k a l i n e
s o l u t i o n s . H a l f - l i f e estimates f o r aqueous s o l u t i o n s
of the n i t r a t e s a l t ranged from more than 3 days at pH
5 , about 11 hours at pH 6 , and 30 minutes at pH 7 . 4 .
A n a l y t i c a l Procedures
In order to study the conversion of Pro-DPH to DPH
i n b i o l o g i c a l systems
d t
fo
h
d
i n the presence of th
were devised to separate the two compounds. This was
followed by a p p l i c a t i o n of the permanganate o x i d a t i o n
technic f o r diphenylmethane d e r i v a t i v e s to form benzophenone, and u l t i m a t e f l u o r o m e t r i c assay of the benzophenone as described by D i l l and Glazko (4_). The sepa
r a t i o n procedure i s o u t l i n e d i n Figure 2.
Blood specimens were c o l l e c t e d with one-tenth v o l
ume of 1 M c i t r a t e b u f f e r added to y i e l d a f i n a l pH of
4 . 5 - 5 . 0 , e f f e c t i v e l y b l o c k i n g f u r t h e r r i n g c l o s u r e of
the remaining Pro-DPH. The blood specimens were
c h i l l e d immediately, c e n t r i f u g e d to separate the p l a s
ma, and assayed the same day. In a t y p i c a l assay, 1 ml
of plasma i s d i l u t e d with 1 ml of water, 0 . 5 g of
NaHCU3 i s added and the mixture i s extracted without
delay by shaking f o r 10 minutes i n a glass-stoppered
t e s t tube with 7 ml 1 , 2 - d i c h l o r o e t h a n e (EDC). Five ml
of the EDC i s t r a n s f e r r e d to a second tube and shaken
f o r 5 minutes w i t h 3 ml 0 . 1 N CHI to separate the ProDPH from the EDC. The HCI e x t r a c t (A) i s r e t a i n e d f o r
f l u o r o m e t r i c assay of Pro-DPH. A p o r t i o n of the r e
maining EDC l a y e r (4 ml) i s t r a n s f e r r e d to another tube
and shaken f o r 5 minutes with 3 ml IN NaOH to e x t r a c t
any DPH. The NaOH e x t r a c t (B) i s a l s o r e t a i n e d f o r
f l u o r o m e t r i c assay of DPH. A l i q u o t s of e x t r a c t s A and
(2 ml) are then made s t r o n g l y a l k a l i n e by the a d d i
t i o n of 1.5 ml of 50% NaOH, 4 ml of -heptane and 300
mg p u l v e r i z e d KMnOij are added, and the mixtures are
heated on a steam bath f o r 30 minutes i n g l a s s - s t o p
pered tubes. The benzophenone produced i n t h i s step i s
extracted i n t o the -heptane l a y e r , a 3 ml p o r t i o n i s
shaken with 1 ml cone, s u l f u r i c a c i d , and the f l u o r e s
cence i s read i n a spectrophotofluorometer at 3 6 0 / 4 9 0
nm as described i n our e a r l i e r report (40 .
Tissue specimens were frozen on blocks of dry i c e
immediately upon removal, weighed, and 10$ (wt/vol)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

185

186

PRO-DRUGS

HN

X c /

NH

li

0
MOLECULAR WEIGHT (SULFATE SALT)

= 837.01

SOLUBILITY (WATER) = 308 MG/ML (pH = 3.3)


DPH EQUIVALENTS =

OF SULFATE SALT.

Figure 1. Chemical structure of Pro-DPH sulfate (N-carbamoyl-2,2-diphenylglycine,2-(diethylamino)ethyl ester, hemisulfate)

1 ml Plasma
+ 0.5 g NaHCO^

+ 7 ml EDC
(10)
5 ml EDC
+ 3 ml 0.1N HCI

( d i s c a r d aqueous) ^ _

(5 )

ml EDC

+ 3 ml IN NaOH

(5)
2 ml HCI

2 ml NaOH

(discard EDC)

PRO-DPH

DPH

FLUOROMETRIC ASSAY VIA BENZOPHENONE


Journal of Clinical Chemistry

Figure 2. Extraction scheme for separation of ProDPH and DPH. The extracts are then subjected to
permanganate oxidation to form benzophenone, which
is assayed by thefluorometrictechnicque. described
elsewhere (4).
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

4.

GLAZKO

ET

AL.

Metabolic Disposition of 5,5-Diphenylhydantoin

187

homogenates were prepared w i t h pH 4 . 5 c i t r a t e b u f f e r


(0.2M). A l l assay r e s u l t s were expressed i n terms of
DPH e q u i v a l e n t s . Fluorometer readings were d i r e c t l y
p r o p o r t i o n a l to concentrations of DPH and Pro-DPH over
a wide range. However, w i t h known standards of DPH and
Pro-DPH, about 5% of the DPH was picked up i n the ProDPH assay, and about 5% of the Pro-DPH was picked up i n
the DPH assay. Nevertheless, t h i s technic provided i n formation which was not a v a i l a b l e i n any other way.
Conversion of Pro-DPH to DPH i n B u f f e r s and i n Plasma
As an example of the a p p l i c a t i o n of t h i s assay
procedure t o . s t a b i l i t studie i v i t r o Pro-DPH s u l
fate i n a f i n a l concentratio
per ml was added to 10 ml pH 7 . 8 phosphate b u f f e r
(0.2M) at 25C and 37C. Volumes of 0 . 5 ml were r e moved by p i p e t t e at frequent i n t r e v a l s , and t r a n s f e r r e d
to 2 . 5 ml volumes of 0.2M c i t r a t e b u f f e r at pH 4 to
stop r i n g c l o s u r e . The r e s u l t s of the assays f o r ProDPH and DPH are shown i n Figure 3- The apparent h a l f l i f e of Pro-DPH at 25C was about 1 hour, whereas at
37C the h a l f - l i f e was only 8 minutes. A s i m i l a r t r i a l
was run w i t h r a t plasma plus pH 7 . 8 phosphate b u f f e r ,
with the r e s u l t s shown i n Figure 4 . Here the apparent
h a l f - l i f e of the Pro-DPH was about 50 minutes at 25C,
and 10 minutes at 37C. DPH l e v e l s showed a c o r r e s ponding i n c r e a s e , i n d i c a t i n g e s s e n t i a l l y complete conv e r s i o n of the Pro-DPH to DPH.
Plasma Levels i n Rats
Albino r a t s r e c e i v i n g intramuscular doses of ProDPH n i t r a t e equivalent to 45 mg DPH per Kg body weight
produced the plasma l e v e l s shown i n Figure 5 The i n i t i a l plasma l e v e l s f e l l o f f q u i t e r a p i d l y , w i t h a h a l f l i f e of about 20 minutes, extending out to 1 hour i n
l a t e r time periods. The i n i t i a l drop i n plasma l e v e l s
appears to be due to r e d i s t r i b u t i o n of the Pro-DPH i n t o
the t i s s u e s , as w e l l as to conversion to DPH by r i n g
c l o s u r e . Assay of the muscle i n j e c t i o n s i t e s at d i f ferent time period a f t e r dosing i n d i c a t e d r a p i d absorpt i o n of Pro-DPH, with removal of 50? of the drug i n
about 15 minutes. The plasma DPH l e v e l s rose r a p i d l y
to a f a i r l y f l a t plateau ( 6 - 8 yg/ml) over the 1 to 4
hour period a f t e r dosing. P a r a l l e l experiments w i t h 45
mg/Kg intramuscular doses of commercially a v a i l a b l e DPH
i n r a t s produced the plasma l e v e l s shown as a broken
l i n e i n Figure 5 Although the plasma l e v e l s of DPH
a f t e r dosing with DPH were somewhat higher than those

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

188

PRO-DRUGS

Figure 3. Conversion of Pro-DPH sulfate to DPH in


pH 7.8 phosphate buffer at 25 and 37 C

Figure 4. Conversion of Pro-DPH sulfate to DPH in rat


plasma buffered to pH 7.8 at 25 and 37C

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

4. GLAZKO ET AL.

Metabolic Disposition of 5,5-Diphenylhydantoin

TIME AFTER DOSING

HOURS

Figure 5. Phsma levels of Pro-DPH and in


rats following intramuscular doses of Pro-DPH ni
trate (solid lines) and DPH (broken line). DPH as
says (), Pro-DPH assays (O). Doses as DPHequivalents per kg body weight were 45 mg/ kg.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

189

190

PRO-DRUGS

r e s u l t i n g from the a d m i n i s t r a t i o n of Pro-DPH, 15 to 30


minutes a f t e r dosing, they f e l l below the DPH l e v e l s
obtained with Pro-DPH i n l a t e r time periods.
Rat plasma l e v e l s r e s u l t i n g from p e r o r a l administ r a t i o n of Pro-DPH s u l f a t e i n doses equivalent to 50 mg
DPH per Kg body weight are shown i n Figure 6. The ProDPH l e v e l s reached a peak at or before the 15 minute
sampling time, and then f e l l r a p i d l y due to t i s s u e d i s t r i b u t i o n and conversion to DPH. The plasma DPH l e v e l s
reached a peak of 8-9 yg/ml 2 hours a f t e r dosing, at a
time when plasma Pro-DPH l e v e l s were q u i t e low. This
could occur only with r e t u r n of DPH to the plasma from
the t i s s u e depots where Pro-DPH had accumulated, thus
p r o v i d i n g a mechanis
extendin
duratio
plasma l e v e l s i n th
at the same dose l e v e l , absorption from the g a s t r o - i n
t e s t i n a l t r a c t was r e l a t i v e l y slow, with considerably
lower plasma l e v e l s (1-2 yg/ml) appearing 8 to 12 hours
a f t e r dosing. Since the plasma h a l f - l i f e of DPH i n
r a t s (< 2 hours) i s much shorter than i n man (> 15
hours), t h i s f a c t o r i s not expected to provide a t h e r a peutic advantage over DPH i n human subjects.
Plasma Levels i n Dogs
Plasma l e v e l s of DPH and Pro-DPH were determined
i n a s e r i e s of dogs r e c e i v i n g intravenous doses of ProDPH s u l f a t e equivalent to 7, l * and 27 mg DPH per Kg
body weight. The r e s u l t s are shown i n Figure 7 The
i n i t i a l plasma l e v e l s of Pro-DPH were considerably
higher than those observed i n the r a t s , due to the d i f ference i n routes of a d m i n i s t r a t i o n . There was an expected r a p i d f a l l i n Pro-DPH plasma l e v e l s accompanied
by a sharp r i s e i n plasma DPH to plateau l e v e l s w i t h i n
1 hour. These were roughly p r o p o r t i o n a l to dosage, and
they were maintained quite w e l l at the two higher dose
l e v e l s over the 160 minute blood sampling p e r i o d . However, at the lower dose l e v e l , there was a n o t i c a b l e
decrease i n the DPH plasma l e v e l s over the same time
period. These doses of Pro-DPH were w e l l t o l e r a t e d by
the dogs, whereas s i m i l a r doses of DPH produced acute
s i d e - e f f e c t s , such as tremors and vomiting. This i s
probably due to the poor d i f f u s i o n of Pro-DPH i n t o the
c e n t r a l nervous system, as i n d i c a t e d by the t i s s u e d i s t r i b u t i o n studies.
1

Tissue D i s t r i b u t i o n Studies
Albino r a t s were given intramuscular doses of ProDPH s u l f a t e equivalent to 50 mg DPH per Kg body weight,

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

GLAZKO ET AL.

Metabolic Disposition of 5,5-Diphenylhydantoin

Figure 6. Plasma levels in rats (two per time pe


riod) following peroral doses of Pro-DPH sulfate
equivalent to 50 mg DPH per kg body weight.
DPH assays (), Pro-DPH assays (Q).

Figure 7. Plasma levels of Pro-DPH and DPH in dogs following intra


venous doses of Pro-DPH sulfate equivalent to 7 (left), 14 (center), and 27
(right) mg DPH equivalents per kg body weight. DPH assays (), ProDPH assays (Q).
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

192

PRO-DRUGS

and the t i s s u e s i n d i f f e r e n t animals were assayed f o r


Pro-DPH and DPH at various time periods a f t e r dosing.
The r e s u l t s are shown i n Figure 8. The Pro-DPH l e v e l s
10 minutes a f t e r dosing were highest i n the kidneys,
lungs and spleen, w i t h p r o g r e s s i v e l y lower l e v e l s occ u r r i n g i n the l i v e r , heart, plasma, s k e l e t a l muscle
and body f a t . The t i s s u e l e v e l s of Pro-DPH f e l l of at
a f a i r l y constant r a t e with an estimated h a l f - l i f e o f
30 minutes. Only t r a c e s of drug were found i n the
b r a i n , i n d i c a t i n g that Pro-DPH does not r e a d i l y cross
the blood-brain b a r r i e r . DPH l e v e l s i n the t i s s u e s
showed a steady increase over the 90 minute period of
the experiment, and i t s d i s t r i b u t i o n was s i m i l a r t o
that observed f o l l o w i n
l
p a r e n t e r a l dose
f
DPH.
A s i m i l a r t i s s u e d i s t r i b u t i o n study was c a r r i e d
out i n a s e r i e s of 5 dogs given intravenous doses o f
Pro-DPH s u l f a t e (equivalent t o 14 mg per Kg body
weight), and s a c r i f i c e d at d i f f e r e n t time periods a f t e r
dosing. The r e s u l t s are shown i n Figure 9 The t i s s u e
d i s t r i b u t i o n of Pro-DPH was very s i m i l a r t o that observed i n the r a t , although i n i t i a l l y at much higher
l e v e l s due t o the route of a d m i n i s t r a t i o n . Most t i s sues had t h e i r highest l e v e l s 5 minutes a f t e r dosing,
i n d i c a t i n g very r a p i d d i f f u s i o n of drug i n t o the t i s sues. The Pro-DPH l e v e l s f e l l r a p i d l y f o r the f i r s t
h a l f - h o u r , and then more slowly with an estimated h a l f l i f e of 35 minutes. Again, only traces of Pro-DPH were
found i n b r a i n (< 1 yg/g), i n d i c a t i n g poor t r a n s p o r t
across the blood-brain b a r r i e r . However, DPH l e v e l s i n
the b r a i n rose to 5-6 yg/g i n about 30 minutes, i n para l l e l w i t h the plasma l e v e l s , i n d i c a t i n g entry o f DPH
i n t o the c e n t r a l nervous system a f t e r i t had been
formed elsewhere i n the body.
Urinary E x c r e t i o n
The u r i n a r y e x c r e t i o n of DPH and Pro-DPH s u l f a t e
was measured i n r a t s f o l l o w i n g I.M. doses of the two
drugs, equivalent t o 50 mg DPH per Kg body weight.
Urine was c o l l e c t e d f o r 22 hours over dry i c e t o keep
the specimens f r o z e n . Upon thawing, pH 4.5 c i t r a t e
b u f f e r was added t o minimize r i n g c l o s u r e . Assay r e s u l t s are shown i n Table 1.
When DPH was administered, the e x c r e t i o n of unchanged DPH represented 0.05-0.15$ of the dose. However, when Pro-DPH was administered, 2.0-3-0$ of the
dose was recovered as f r e e DPH. Since very l i m i t e d
q u a n t i t i e s o f DPH appear i n the u r i n e f o l l o w i n g admini s t r a t i o n of DPH, i t i s d i f f i c u l t t o see why such i n -

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

4. GLAZKO ET AL.

Metabolic Disposition of 5,5-Diphenylhydantoin

100

<

PRO-DPH ASSAY

DPH ASSAY

10

'-

F /

0.1

V
10

30

90
10
TIME AFTER DOSING

30
MINUTES

KIDNEY
SPLEEN
LUNG
LIVER

FAT
BRAIN
90

Figure 8. Tissue levels of Pro-DPH and DPH in rats


following intramuscuhr doses of Pro-DPH sulfate equiva
lent to 50 mg DPH per kg body weight

TIME AFTER DOSING - MINUTES

Figure 9. Tissue levels of Pro-DPH and DPH in dogs


following intravenous doses of Pro-DPH sulfate equiva
lent to 14 mg DPH per kg body weight
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

193

194

PRO-DRUGS

TABLE 1
ASSAY OF 22-HOUR RAT URINE AFTER A SINGLE 50 MG/KG
INTRAMUSCULAR DOSE OF PRO-DPH OR DPH

DRUG
ADMINISTERED

URINARY EXCRETION
PER CENT OF DOSE

RAT
NO.
PRO-DPH

DPH

TOTAL

(*)

{%)

(%)

PRO-DPH

k.2

2.0

6.2

DPH

0.15

0.15

0.05

0.05

creased amounts are excreted f o l l o w i n g a d m i n i s t r a t i o n


of Pro-DPH, unless the Pro-DPH i t s e l f undergoes r i n g
closure i n urine f i l t r a t e s .
The assays i n d i c a t e that
some Pro-DPH i s excreted unchanged i n r a t u r i n e .
This
aspect of Pro-DPH metabolism requires f u r t h e r i n v e s t i g a t i o n , since i t i s p o s s i b l e that the DPH generated
from Pro-DPH i n urine may reach a s a t u r a t i o n p o i n t ,
producing c r y s t a l l u r i a or even d e p o s i t i o n of c r y s t a l s
i n the r e n a l t i s s u e . The pH of the urine would be expected t o be a c r i t i c a l f a c t o r i n a f f e c t i n g the r a t e of
r i n g c l o s u r e . These and other r e l a t e d aspects o f ProDPH metabolism r e q u i r e f u r t h e r examination i n animal
s t u d i e s , e s p e c i a l l y with repeated doses of Pro-DPH, before t h i s pro-drug can be regarded as a s u i t a b l e candidate f o r therapeutic use i n man.
Summary
Pro-DPH i s of i n t e r e s t because of i t s high water
s o l u b i l i t y and ease of conversion to DPH i n b i o l o g i c a l
systems. Assay procedures were devised to measure the
concentration of DPH and Pro-DPH i n the same b i o l o g i c a l
specimens. Pro-DPH was found t o be absorbed r a p i d l y
from intramuscular i n j e c t i o n s i t e s i n r a t s and dogs,
and i t showed good absorption c h a r a c t e r i s t i c s from the
g a s t r o - i n t e s t i n a l t r a c t of the r a t . The plasma l e v e l s
of Pro-DPH f e l l r a p i d l y due to d i s t r i b u t i o n i n t o the
t i s s u e s and conversion t o DPH by r i n g c l o s u r e .
This
was accompanied by the appearance of high and extended
plasma l e v e l s of DPH, which were roughly p r o p o r t i o n a l

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

4. GLAZKO ET AL.

Metabolic Disposition of 5,5-Diphenylhydantoin

to dosage. Highest concentrations of Pro-DPH were


found in the kidneys, with somewhat lower concentra
tions in the spleen and lungs; only traces of Pro-DPH
were found in the brain. Excretion data in rats indi
cate that some Pro-DPH is excreted unchanged in the
urine, and that a portion may be converted to DPH
after clearance through the kidneys. Additional work
is needed in this area before Pro-DPH can be cleared
for human t r i a l .
Literature Cited
1. Kozelka, F. L . , and Hine, C. E . , J. Pharmacol.
Exper. Therap. (1941) 72 276
2. Stella, V . , an
(1973), 62, 962.
3. Glazko, A. J., Dill, W. ., Wheelock, R. .,
Nemanich, ., Croskey, L . , and Higuchi, T., Feder
at. Proc. (1973), 32, 684.
4. D i l l , W. ., and Glazko, A. J., Clin. Chem. (1972),
18, 675.
FOOTNOTE
Dilantin is the Parke, Davis & Company tradename for
5,5-diphenylhydantoin.
*

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5
Case Histories of the Development of Pro-drugs for
Use in the Formulation of Cytotoxic Agents in
Parenteral Solutions
ARNOLD J. REPTA
Department of Pharmaceutical Chemistry, School of Pharmacy,
University of Kansas, Lawrence, Kans. 66044
Introduction
The pro-drug approac
drug delivery through a number of related but distinctly different mechanisms. These might include use of a
pro-drug in attempting 1) to alter the rate of metabolism of the drug, 2) to alter the rate of release from
a dosage form or depot site, and/or 3) to deliver the
drug to a specific site or organ in the body. The
achievement of any one or a combination of two or more
of these objectives would be expected to have significant effects on the concentration vs time profile of
the drug at the receptor site. Although the technology
for achieving any one or more such goals could probably
be developed for a given drug substance, a good deal of
data would have to be available relative to the metabolism, site of action, toxicity and desired concentration-time profile for that drug in the body. Only on
the basis of such information would i t be possible to
define the specific properties and characteristics that
need be incorporated in the pro-drug. Obviously, the
application of pro-drug approaches in order to attain
such objectives requires that there be both a great
deal of interest in the drug and considerable advantage
to be gained before one could justify committing the
resources necessary for the design and development of a
suitable pro-drug.
A more common and presumably more achievable objective, which would involve the use of pro-drugs for
improved drug delivery, would be that concerned with the
enhancement of the absorption or bioavailability of a
drug through some reversible alteration of the properties of the drug. Pro-drugs used for such a purpose
would normally exhibit an apparent increase in the solubility or stability of the drug which in turn would
196
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

REPTA

Cytotoxic Agents in Parenteral Solutions

197

allow the drug to be released from the dosage form and


to a r r i v e at the receptor s i t e more r a p i d l y and/or more
completely when employed as the pro-drug form.
The problem of poor b i o a v a i l a b i l i t y of c e r t a i n cyt o t o x i c agents of i n t e r e s t i n cancer chemotherapy has
apparently l e d to a d e s i r e by i n v e s t i g a t o r s working
with the N a t i o n a l Cancer I n s t i t u t e to carry out c l i n i c a l t e s t i n g of such agents as intravenous s o l u t i o n s .
This preference f o r s o l u t i o n dosage forms r e s u l t s from
the current acceptance of intravenous a d m i n i s t r a t i o n of
a drug as r e p r e s e n t i n g complete b i o a v a i l a b i l i t y ( 1 ) .
While such equivalence may be argued (1,2), comparison
of c l i n i c a l a c t i v i t y of d i f f e r e n t drugs and drug r e gimes administered i t h i
would c e r t a i n l
to be superior to a d m i n i s t r a t i o
combination of routes.
Although the use of intravenous s o l u t i o n s i n c l i n i c a l t e s t i n g of c y t o t o x i c agents seems to be both r a t i o n a l and w e l l accepted, often there are major problems encountered i n the formulation of some of these
agents i n s o l u t i o n s s u i t a b l e f o r intravenous use. Such
problems u s u a l l y i n v o l v e the s o l u b i l i t y and/or the
chemical s t a b i l i t y of the candidate drugs i n p h y s i o l o g i c a l l y acceptable solvent systems. Many of these spec i f i c problems may be overcome by formulating as a c i d i c
or a l k a l i n e aqueous s o l u t i o n s , by using mixed solvent
systems, by using dry formulations which are r e c o n s t i tuted j u s t p r i o r to use, and by various other completel y acceptable techniques. However, on occasion c e r t a i n
c y t o t o x i c agents present problems f o r which such s o l u t i o n s are not adequate and these are the types of substances f o r which the pro-drug approach i s most a p p l i cable.
The i d e n t i f i c a t i o n and use of a pro-drug system as
the s o l u t i o n to a d e l i v e r y problem which could be
solved by s a l t formation, pH adjustment, use of c o s o l vents, or other simple formulation techniques appears
to be a superfluous exercise and should probably be
discouraged i n l i g h t of the time and resources which
would normally be required to b r i n g such an approach to
a s u c c e s s f u l conclusion. Thus when a s t a b i l i t y or s o l u b i l i t y problem has been encountered, the r a t i o n a l development of the answer to the problem begins w i t h a
s e r i e s of questions which must be asked and answered
carefully.
The f i r s t question to be considered i s , "Can the
gross problem be obviated by any of the more or l e s s
t r i v i a l manipulations such as pH adjustment, e t c ? " I f
the answer to the f i r s t question i s n e g a t i v e , the second question i s asked. I t i s , "What are the basic f a c -

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

198

PRO-DRUGS

t o r s g i v i n g r i s e t o the gross problem being confronted?" The answer t o t h i s question i s u s u a l l y m u l t i f a c eted and often cannot be answered with complete cert a i n t y . Usually the answer requires o b t a i n i n g and
e v a l u a t i n g various p h y s i c a l and chemical data about the
candidate drug. The p a r t i c u l a r nature of the data
needed depends on the problem type. For i n s t a n c e , i f
the problem i s one of the low aqueous s o l u b i l i t y of the
substance, then information on the melting behavior,
the s o l u b i l i t y i n a v a r i e t y o f s o l v e n t s , the s t r u c t u r a l
f e a t u r e s , e t c . , are important.
I f the problem i s s t a b i l i t y r e l a t e d , then the nature of the degradation
products, the k i n e t i c order of the degradation process,
information r e l a t i v t r e a c t i o mechanis
d
d
a l l other data p e r t a i n i n
are u s e f u l . The c a r e f u
type
information and s i m i l a r information on r e l a t e d compounds often allows one t o s p e c i f y with some degree of
c e r t a i n t y the basic f a c t o r s causing the problem. For
example, the low aqueous s o l u b i l i t y of a r a t h e r p o l a r
compound which has a high melting point and a low s o l u b i l i t y i n v i r t u a l l y a l l solvents could perhaps be a t t r i b u t e d t o strong i n t e r m o l e c u l a r hydrogen bonding i n t e r a c t i o n s i n the c r y s t a l l i n e s t a t e . This would cause
the thermodynamic a c t i v i t y of the solute phase to be
low which i n t u r n gives r i s e t o the observed low s o l u b i l i t y . A second example, i n v o l v i n g a s t a b i l i t y problem, might be a s i t u a t i o n where the data may suggest
that i n t e r - or i n t r a m o l e c u l a r c a t a l y s i s by some funct i o n a l group i n the drug molecule i s responsible f o r
the r a p i d degradation.
A f t e r answering the second question, one can proceed to the next question which i s , "What s p e c i f i c type
of manipulation of the molecule or system can be accomp l i s h e d so that the basic cause or causes w i l l be suff i c i e n t l y ameliorated so as t o solve the gross problem?"
Normally the generic answer t o t h i s question i s
not a d i f f i c u l t one. This i s p a r t i c u l a r l y true i f the
previous question ( r e l a t i v e t o the basic causes of the
problem) was answered i n unequivocal terms. As an example, a s o l u t i o n t o the above described s o l u b i l i t y
problem, which stemmed from hydrogen bonding, might be
the d i s r u p t i o n of the hydrogen bonding by preparing a
chemical d e r i v a t i v e which would be incapable of a c t i n g
as a hydrogen donor. S i m i l a r l y an answer to the s t a b i l i t y problem p r e v i o u s l y described might i n v o l v e using
some p h y s i c a l or chemical a l t e r a t i o n of the system
which would t i e up the f u n c t i o n a l group responsible f o r
the c a t a l y s i s of the decomposition r e a c t i o n .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

REPTA

Cytotoxic Agents in Parenteral Solutions

199

The f o u r t h and f i n a l (and perhaps the most d i f f i c u l t ) question to be asked i s , "What s p e c i f i c pro-drug
approach can be used to achieve the desired e f f e c t on
the p r o p e r t i e s of the drug substance and y e t , while
being s u f f i c i e n t l y stable i n the dosage form, w i l l rapi d l y release the parent drug i n the body upon administ r a t i o n ? " The answer to t h i s question i s often a d i f f i c u l t one since i t r e q u i r e s that the approach to be
used must s a t i s f y the general aims of the answer to
question three and a d d i t i o n a l l y i t must contain a
b u i l t - i n r e l e a s i n g mechanism which i s t r i g g e r e d by some
agent or event encountered upon a d m i n i s t r a t i o n of the
pro-drug. This t r i g g e r must dependently and r a p i d l y
allow or cause the pro-drug to r e v e r t to the free par
ent drug.
I t appears that there are at l e a s t three such po
t e n t i a l t r i g g e r i n g mechanisms which are encountered i n
the a d m i n i s t r a t i o n of intravenous s o l u t i o n s . The f i r s t
of these i s the various enzymes i n the blood and other
t i s s u e s (3.) I f one designs a pro-drug which w i l l
serve as a substrate f o r one or more of the enzymes
found i n the body and i f t o t a l enzymatic a c t i v i t y i s
s u f f i c i e n t l y l a r g e , then the release of the drug from
the pro-drug w i l l be much enhanced.
Another p o t e n t i a l t r i g g e r i s the event i n v o l v i n g
the d i l u t i o n of the pro-drug formulation upon administ r a t i o n of the s o l u t i o n i n t o the bloodstream. Such d i l u t i o n r e s u l t s i n reduced concentrations of components
and thus causes s h i f t s i n e q u i l i b r i a . An example of a
pro-drug system f o r which t h i s t r i g g e r would be s u i t able i s one i n v o l v i n g complexation which i s subject to
the law of mass a c t i o n (4_).
The buffered and r e l a t i v e l y constant value of the
p h y s i o l o g i c a l pH (= 7 . 4 ) i s a t h i r d f a c t o r which may be
u s e f u l i n t r i g g e r i n g the release of a drug from a prodrug. In those cases where the pH and b u f f e r capacity
of the blood i s to be used to advantage, i t would seem
to be necessary that the p r o - d r u g K i r u g r e a c t i o n be
very pH s e n s i t i v e , and that the pro-drug formulation
used be at a pH value appreciably d i f f e r e n t than
pH 7 . 4 .
In view of the l i m i t e d number of t r i g g e r i n g mechanisms a v a i l a b l e one must c a r e f u l l y consider the p h y s i co-chemical c h a r a c t e r i s t i c s of the v a r i o u s p o t e n t i a l
pro-drug approaches w i t h a view toward s e l e c t i n g only
those pro-drug systems which w i l l be most responsive to
one or more of the a v a i l a b l e t r i g g e r s encountered upon
intravenous a d m i n i s t r a t i o n of the f i n a l formulation.
When a l l of the above four questions have been
s a t i s f a c t o r i l y answered, the pro-drug system must be
f

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

200

PRO-DRUGS

prepared and t e s t e d . I n i t i a l i n v i t r o t e s t s w i l l usu


a l l y screen out many unsuccessful approaches. However,
the f i n a l proof of the u t i l i t y of the pro-drug approach
s e l e c t e d w i l l n e c e s s a r i l y await the r e s u l t s of i n vivo
testing.
The remainder o f t h i s d i s c u s s i o n i s a review of
the concerns involved i n the i d e n t i f i c a t i o n and evalua
t i o n of some pro-drug approaches used i n attempting t o
solve problems associated with the intravenous d e l i v e r y
of three c y t o t o x i c agents.
Case I . I d e n t i f i c a t i o n o f 9-(5-0-formyl-g-D-arabinofuranosyl)adenine as a pro-drug o f l-g-D-arabinofuranosyladenine.
The compound --D-arabinofuranosyladenine ( I ) i s
a nucleoside which i s most commonly c a l l e d ara-. I t
i s the 2 epimer of adenosine ( I I ) and considerable i n
t e r e s t has been generated i n the compound because of
i t s a b i l i t y t o i n h i b i t the growth of some v i r u s e s (5.)
and tumors ( 6 ) .
f

OH
I
ara~A

OH

OH
II
adenosine

Some c l i n i c a l t e s t i n g of ara- as a cancer chemotherapeutic agent has been attempted but a f u l l scale
e v a l u a t i o n has been severely l i m i t e d because of prob
lems associated with the a d m i n i s t r a t i o n of the drug.
Human adult doses of 2.5 grams have been suggested and
a c t u a l l y administered as an aqueous intravenous i n f u
s i o n , but the low s o l u b i l i t y value of ^0.5 mg per ml of
water (8_) found f o r ara- have n e c e s s i t a t e d the use of
5 t o 6 l i t e r s of such s o l u t i o n s . Obviously, such a
large volume of s o l u t i o n r e q u i r e s a d m i n i s t r a t i o n over a
prolonged period of time ( 9 ) .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

REPTA

Cytotoxic Agents in Parenteral Solutions

201

Along w i t h the n e c e s s a r i l y slow a d m i n i s t r a t i o n ,


there i s a problem of the enzymatic metabolism of ara-A
by adenosine deaminase (10,11,) to the corresponding hypoxanthine d e r i v a t i v e ( I I I , ara-) which i s i n e f f e c t i v e
i n i n h i b i t i n g tumor growth (12.). The b i o l o g i c a l h a l f l i f e of the enzymatic deamination r e a c t i o n i s reported
to be about 30 minutes (12). Such f a s c i l e metabolism,
together w i t h the slow a d m i n i s t r a t i o n , makes i t impos
s i b l e to achieve appreciable blood l e v e l s of ara-A.
Thus a meaningful c l i n i c a l e v a l u a t i o n of the inherent
antitumor a c t i v i t y of ara-A was d i f f i c u l t to make.

III
ara-H

IV
ara-A-5 -formate
1

I t seemed that the most l o g i c a l and simplest ap


proach to achieving improved drug d e l i v e r y i n t h i s case
was to increase the r a t e at which the drug could be ad
m i n i s t e r e d . This i n v o l v e d preparing and using a more
concentrated s o l u t i o n s u i t a b l e f o r intravenous use.
Rather extensive studies aimed at producing an i n
creased apparent s o l u b i l i t y of ara-A through the use of
mixed solvent systems, agents which would i n h i b i t c r y s t a l i z a t i o n , and other approaches had been c a r r i e d out
(13) and proved f r u i t l e s s . The weakly basic nature of
ara-A (pKa ^ 3.7 (lJO) together with i t s low aqueous
s o l u b i l i t y and high dose r u l e d out the use of an a c i d
s a l t to overcome the s o l u b i l i t y problem. Thus, i t ap
peared that an increase i n the apparent s o l u b i l i t y
could not be achieved by simple p h y s i c a l approaches and
the p o s s i b i l i t y of u t i l i z i n g some s u i t a b l e pro-drug
systems began to be considered.
A comparison of the s t r u c t u r a l features and some
of the p r o p e r t i e s of ara-A and adenosine, as shown i n
Table I , brought out some u s e f u l p o i n t s . F i r s t of a l l ,

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

202

PRO-DRUGS

Table I - Some P r o p e r t i e s of Adenosine,


Ara-A and Ara-A-5 -Formate (14)
Water s o l u b i l i t y ,
mp
mol. wt.
(25) M
!

adenosine
ara-a
ara-A-5 -formate
1

*
^
^

0.02
0.0018
0.12a

235
^ 260
* 175

267
267
295

Due to h y d r o l y s i s of the e s t e r t h i s i s an approximate


value.

both ara-A and adenosine are h i g h l y p o l a r compounds and


capable of i n t e r a c t i n s t r o n g l w i t h wate
t
p o s i t i o n s , yet n e i t h e
both have r a t h e r high melting points, and t h e i r s o l u b i l
i t i e s i n most solvents i s quite low. Furthermore, a l though ara-A and adenosine are c l o s e l y r e l a t e d s t r u c t u r a l l y , i t i s noteworthy that adenosine i s more than
ten times more s o l u b l e than ara-A. The conclusions
that were reached on the b a s i s of such information was
that the low s o l u b i l i t y of these nucleosides was due to
strong i n t e r m o l e c u l a r i n t e r a c t i o n s i n the c r y s t a l l i n e
s t a t e which r e s u l t e d i n a very low thermodynamic a c t i v i t y f o r the s o l u t e phase. On the basis of the s t r u c t u r a l features of the compounds, these strong i n t e r a c t i o n s appeared to be due most probably to hydrogen
bonding between the various proton donor and acceptor
s i t e s i n the molecules. The order of magnitude d i f f e r ence i n s o l u b i l i t y between the two epimers f u r t h e r suggested that the extent of hydrogen bonding changed app r e c i a b l y w i t h only moderate changes i n molecular geometry. Thus i t appeared that i f i t were p o s s i b l e to
decrease the i n t e r m o l e c u l a r hydrogen-bonding i n the
c r y s t a l l i n e ara-A, while not s i g n i f i c a n t l y decreasing
the o v e r a l l a b i l i t y of the molecule to i n t e r a c t w i t h
water, i t should be p o s s i b l e to increase s u b s t a n t i a l l y
the apparent aqueous s o l u b i l i t y . I t was a n t i c i p a t e d
that a decrease i n hydrogen bonding could most e a s i l y
be accomplished by decreasing the hydrogen donor capaci t y of the arabinose p o r t i o n of the molecule by s u b s t i t u t i n g non-donor groups f o r the hydrogen atoms.
At t h i s point i n the study i t becomes necessary to
attempt to define r a t h e r s p e c i f i c a l l y a chemical d e r i v a t i v e of ara-A which might be used as a s o l u b l e prodrug. On the b a s i s of the c o n s i d e r a t i o n s which f o l l o w ,
the 5'formate e s t e r of ara-A (IV) was i n i t i a l l y s e l e c t ed ( 8 ) .
An e s t e r d e r i v a t i v e was chosen since e s t e r i f i c a t i o n of a hydroxy group would e l i m i n a t e the hydrogen-

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

REPTA

Cytotoxic Agents in Parenteral Solutions

203

donor capacity at that p o s i t i o n . Although an ether der i v a t i v e would a l s o be e f f e c t i v e i n e l i m i n a t i n g the hydrogen donor a c t i v i t y , the ester was p r e f e r e n t i a l l y seLected because of i t s greater ease of h y d r o l y s i s , esp e c i a l l y i n the presence of the various esterases which
abound i n blood and other t i s s u e s (3.).
Such enzymatic
assistance could therefore serve as the t r i g g e r i n the
release of the drug from the pro-drug.
The choice of the 5 hydroxyl as the s i t e of
e s t e r i f i c a t i o n was based on the general knowledge that
esters of primary a l c o h o l s are normally more e a s i l y hydrolyzed than the corresponding e s t e r s of secondary a l cohols such as are found at the 2 and 3 p o s i t i o n s of
ara-A. An a d d i t i o n a l
fo desirin
derivativ
of the 5 hydroxyl wa
d e r i v a t i v e s of compound
poo
substrates f o r adenosine deaminase (.6,10.) and thus the
5 e s t e r of ara-A would probably not undergo appreciable metabolism p r i o r to h y d r o l y s i s . While t h i s i s an
added bonus i t i s not an e s s e n t i a l f a c t o r i n so f a r as
achieving an increased molar s o l u b i l i t y i s concerned.
The choice of the formate e s t e r r a t h e r than some
other e s t e r was made on the b a s i s of the compact and
r e l a t i v e l y p o l a r nature of the formate group which was
not expected to n o t i c e a b l y a l t e r the o v e r a l l h y d r o p h i l i c character of the pro-drug r e l a t i v e to the parent
compound. The f a c t that formate e s t e r s are normally
more r a p i d l y hydrolyzed than corresponding homologs was
a l s o important ( 1 5 ) .
The reasons f o r choosing the monoformate e s t e r i n
preference to the diformate or t r i f o r m a t e esters
stemmed mainly from a d e s i r e to use as simple a system
as p o s s i b l e i n order to be able to f o l l o w u l t i m a t e l y
the regeneration of the drug. I f the t r i e s t e r was to
be used as the pro-drug, ( p r o v i d i n g i t was s u f f i c i e n t l y
s o l u b l e ) random h y d r o l y s i s of the esters would potent i a l l y r e s u l t i n three d i f f e r e n t diformate esters which
i n t u r n would hydrolyze to y i e l d three d i f f e r e n t monoe s t e r s . This k i n e t i c sequence would then r e s u l t i n the
simultaneous existence of s e v e r a l pro-drug species
which would be present i n amounts r e l a t e d to t h e i r
r a t e s of formation and degradation.
The s i t u a t i o n i s
somewhat simpler f o r the diformate d e r i v a t i v e s , but
s t i l l p o t e n t i a l l y q u i t e complicated when compared to
that to be expected with the 5 monoformate.
Ara-A-5'-formate was prepared (8_) and the aqueous
s o l u b i l i t y was found to change i n the a n t i c i p a t e d d i f

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

204

PRO-DRUGS

r e c t i o n . As seen i n Table I , the s o l u b i l i t y increase


was accompanied by a decrease i n the melting point i n d i c a t i n g a r e d u c t i o n i n the i n t e r m o l e c u l a r i n t e r a c t i o n s
i n the pro-drug e s t e r r e l a t i v e to c r y s t a l l i n e ara-A.
The approximately 65 f o l d increase i n the molar s o l u b i l i t y of the pro-drug i n comparison to ara-A meant
that the volume of intravenous s o l u t i o n necessary to
d e l i v e r the equivalent of 2 . 5 grams of ara-A could be
decreased from 5 or 6 l i t e r s to l e s s than 100 ml. Obv i o u s l y such a r e d u c t i o n i n volume would allow the drug
to be administered i n a much shorter p e r i o d of time.
Although the problem of the low s o l u b i l i t y of a r a A could obviously be solved by using a r a - A - 5 - f o r m a t e ,
the p o t e n t i a l u t i l i t
pro-drug of ara-A depende
i n b i o l o g i c a l f l u i d s and i t s s t a b i l i t y i n an aqueous
s o l u t i o n s u i t a b l e f o r intravenous use. In v i t r o studi e s c a r r i e d out i n 91% whole human blood at 3 7 (jB)
showed h y d r o l y s i s of the ara-A-5'-formate, to y i e l d
ara-A, occurred r a p i d l y . Data from one such run i s
shown i n Figure 1 where i t can be seen that h y d r o l y s i s
of ara-A-5'-formate to ara-A was e s s e n t i a l l y complete
i n about 30 minutes. The r a t h e r r a p i d disappearance of
the regenerated ara-A i s due presumably to i t s metabol i s m by adenosine deaminase ( , 1 0 ) . Nevertheless peak
l e v e l s of ara-A corresponding t o between 60% and 70$
of the administered dose were achieved. Since both the
r a p i d h y d r o l y s i s of the e s t e r and the deamination of
ara-A are r e a c t i o n s which are enzyme a s s i s t e d , i t i s
not s u r p r i s i n g that the r a t e s of both processes were
found to be somewhat concentration dependent (80.
The antitumor a c t i v i t y of ara-A-5 -formate, when
t e s t e d i n t r a p e r i t o n e a l l y i n mice innoculated with e i ther Heptoma 134 or E h r l i c h a s c i t e s tumor c e l l s , was
found to be equivalent to that of ara-A ({$) suggesting
that ara-A was regenerated i n v i v o .
The remaining question which had yet to be answered dealt with the s u i t a b i l i t y of the proposed prodrug i n a formulation acceptable f o r intravenous admini s t r a t i o n . The aqueous s t a b i l i t y of ara-A-5'-formate
was studied and i t was found, not s u r p r i s i n g l y , that
the most serious degradation problem involved hydrolys i s of the formate e s t e r to y i e l d ara-A (8). The e f f e c t
of pH on the r a t e of the h y d r o l y t i c r e a c t i o n was studi e d at 25 (<B) and a p o r t i o n of that p r o f i l e obtained
i s shown i n F i g . 2 . At the p h y s i o l o g i c a l pH of 7 . 4 , the
observed r a t e was ^ 3 x 10~3 m i n u t e ^ which corresponds
1

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5. REPTA

Cytotoxic Agents in Parenteral Solutions

205

to a h a l f - l i f e of about 4 hours. At pH 4 . 2 to 4 . 5 ,
which was the pH range of greatest s t a b i l i t y , the observed h a l f - l i f e was approximately 1 0 days.
Although i t was obvious from the data that the
pro-drug could not be formulated as an aqueous s o l u t i o n with acceptable long term s t a b i l i t y , the formate

TIME (minutes)
Journal of Pharmaceutical Sciences

Figure 1. Plot of the concentrations of ara-A-5'-formate and


ara-A in 91% whole blood at 37 as a function of time, (8).
Initial concentration of ara-A-5'-formate was 2 mg/ml.

ester was s u f f i c i e n t l y (8) s t a b l e at pH 4 . 5 to be used


i n a l y p o h i l i z e d formulation to be r e c o n s t i t u t e d at the
time of use.
Such r e c o n s t i t u t e d s o l u t i o n s , c o n t a i n i n g the
equivalent of 3 0 mg of ara-A/ml (as the pro-drug) would
not be expected to undergo s u f f i c i e n t h y d r o l y s i s i n a
5 to 6 hour p e r i o d to produce a saturated s o l u t i o n of

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

206

PRO-DRUGS

ara-A and thus there would be no problem of p r e c i p i t a


t i o n during the time necessary to administer the pro
drug formulation. Thus i t appears i n the case of a r a A that the ara-A-5'-formate meets v i r t u a l l y a l l the
c r i t e r i a r e q u i r e d of a pro-drug s u i t a b l e f o r use i n a
intravenous f o r m u l a t i o n .

41
3

PH
Journal of Pharmaceutical Sciences

Figure 2. Partial profile of the effects of pH on the observed


first order rate constant for the spontaneous hydrolysis of araA-5'-formate at 25 (8)

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

REPTA

Cytotoxic Agents in Parenteral Solutions

207

Case I I . Hexamethylmelamine-Gentisic A c i d Complexes as


a Soluble Pro-Drug form of Hexamethylmelamine.
Hexamethylmelamine (V) i s a c y t o t o x i c agent which
has commanded c l i n i c a l i n t e r e s t p r i m a r i l y because of
the c o n s i s t e n t , a l b e i t low, response rates obtained
when i t has been used i n the treatment of s e v e r a l s o l i d
tumors i n c l u d i n g lung cancers ( 1 6 , 1 7 ) *

V
Hexamethylmelamine

VI
G e n t i s i c Acid

The d e s i r e f o r an intravenous formulation of t h i s


drug stemmed from the occurance of the side e f f e c t s
( p r i m a r i l y nausea and vomiting) which i t s use i l l i c i t e d
when given o r a l l y (1_8). P r i o r attempts had been made
to provide a s u i t a b l e intravenous preparation based on
the use of an a c i d i c s o l u t i o n of the weakly basic drug.
However, the combined e f f e c t s of a pK, = 9 - 1 0 , an aqueous s o l u b i l i t y of approximately 0 . 1 mg per ml (at room
temperature) (19.) and an estimated dose of about 100 to
200 mg (18) r e s u l t e d i n an aqueous formulation of pH 2 .
The use of t h i s preparation caused serious problems of
l o c a l i r r i t a t i o n and thrombophlebitis (18) which may
have been due to e i t h e r the a c i d i t y of the s o l u t i o n or
an inherent property of the drug or a combination of
both.
In attempting to e l i m i n a t e or at l e a s t reduce the
side e f f e c t s of intravenous a d m i n i s t r a t i o n , a l e s s
a c i d i c aqueous s o l u t i o n (pH 3 . 5 or greater) c o n t a i n i n g
hexamethylmelamine at a concentration of at l e a s t 3 mg
per ml was requested (1_8). Because of i t s somewhat low
s o l u b i l i t y i n most solvents (see Table I I ) which might
be used as cosolvents together with water, the use of a
s u i t a b l e cosolvent mixture d i d not appear to o f f e r a
good p r o b a b i l i t y of success. The f a c t s that the compound e x h i b i t s good s o l u b i l i t y i n non-polar solvents

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

208

PRO-DRUGS

Table I I - Approximate S o l u b i l i t y of Hexamethylmelamine


i n Various Solvents at Ambient Temperature
(19)
Solvent
Approximate s o l u b i l i t y (mg/ml)
?

Water
,-Diethylacetamide
Dimethylsulfoxide
Ethanol
Ethylacetate
Propylene g l y c o l
Polyethylene g l y c o l 400
Chloroform
Benzene
D i e t h y l ether

'V/O.l
30-50
a>5
^15
*15

<2
<2
^200
^10 6

such as benzene, chloroform and d i e t h y l ether, while


being considerably l e s s soluble i n non-polar solvents
such as propylene g l y c o l , ethanol and e s p e c i a l l y water,
appeared to suggest that the s o l u b i l i t y problem i n t h i s
case was not so much due t o a low a c t i v i t y of the mole
cules i n the s o l i d solute as i t was to a r a t h e r high
a c t i v i t y of the d i s s o l v e d solute i n the p o l a r s o l v e n t s .
Thus one approach to i n c r e a s i n g s o l u b i l i t y was t o de
crease the a c t i v i t y of the solute i n s o l u t i o n by some
means.
A method of achieving such a decrease of a c t i v i t y
i n an aqueous s o l u t i o n would be through the use of a
more h y d r o p h i l i c chemical d e r i v a t i v e . However, inspec
t i o n of the chemical s t r u c t u r e suggested that the more
apparent chemical d e r i v a t i v e s capable of achieving the
s o l u b i l i t y i n c r e a s e , such as quaternary s a l t s , probably
would not be u s e f u l as pro-drugs since they would not
be expected t o r e a d i l y release the drug under b i o l o g i
cal conditions.
The apparent l a c k of chemical s i t e s s u i t a b l e f o r
forming a p o t e n t i a l l y u s e f u l chemically derived pro
drug r e s u l t e d i n the c o n s i d e r a t i o n of complexation as
another method of reducing the a c t i v i t y of the d i s
solved hexamethylmelamine. A number of c h a r a c t e r i s t i c s
of the hexamethylmelamine molecule, i n c l u d i n g i t s aro
matic and i t s weakly basic nature, suggested that i t
would associate or complex with a s u i t a b l e l i g a n d . In
view of the f a c t that i n the pH 3.5 to 4.0 r e g i o n , the
apparent increase i n s o l u b i l i t y which was needed was
only about 5 to 10 f o l d , i t seemed l i k e l y that t h i s
magnitude of increase could be achieved through com
p l e x a t i o n . I f the s o l u b i l i t y increase d e s i r e d had been
2 or 3 orders of magnitude or greater, complexation
would not have been s e r i o u s l y considered, since such

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

REPTA

Cytotoxic Agents in Parenteral Solutions

209

large increases are not o f t e n achieved by a s s o c i a t i o n


of organic molecules i n aqueous media.
In choosing a p o t e n t i a l l i g a n d f o r the complexat i o n of hexamethylmelamine two f a c t o r s had to be considered. One of these was a c c e p t a b i l i t y of the l i g a n d
from the p h y s i o l o g i c a l standpoint. The other was the
a b i l i t y of the agent t o i n t e r a c t with the drug. Somewhat f o r t u i t o u s l y gntisic a c i d (VI) was the l i g a n d
i n i t i a l l y chosen. This choice was based on the f a c t s
that i t i s r e l a t i v e l y non-toxic (2), has been used
m e d i c i n a l l y ( 21) , and thus appeared to be acceptable
from the b i o l o g i c a l standpoint. Through experience
gained from other studies of the complexation of gent i s i c a c i d w i t h heteraromati
such as c a f f e i n e (22),
a c i d or the gentisate i o n might be l i k e l y to i n t e r a c t
with a compound such as hexamethylmalamine under s u i t able c o n d i t i o n s . Thus the complexation between g e n t i s i c a c i d and hexamethylmelamine was studied and f o r t u n a t e l y , as w i l l be discussed below, the system achieved
the d e s i r e d r e s u l t s (23).
Before proceeding, a b r i e f d i s c u s s i o n of complexat i o n and some advantages of such a pro-drug system i n
the intravenous a d m i n i s t r a t i o n of drugs i s i n order.
Complexation normally r e f e r s to the a s s o c i a t i o n of two
or more molecules of two d i f f e r e n t compounds i n a r e v e r s i b l e manner (2_4). The e q u i l i b r i u m involved i n the
formation of a p a r t i c u l a r complex species may be w r i t en as

( e q

1 )

mS + nL

In eq. 1, S and L represent the i n t e r a c t i n g mole


c u l a r s p e c i e s , one of which i s normally c a l l e d the sub
s t r a t e and the other the l i g a n d . S L represents the
complex species formed and m and are the c o e f f i c i e n t s
which express the stoichiometry of the complex.
The e q u i l i b r i u m involved may be defined by an as
s o c i a t i o n constant which may be w r i t t e n i n terms of
the concentration of the complexed and uncomplexed spe
c i e s as

= [S L ]/[S] [L]
m

(eq. 2 ) .

The p r o p e r t i e s of the substrate and l i g a n d molecules i n


the complex S L a r e often quite d i f f e r e n t from (and
r a t h e r independent o f ) those of the uncomplexed spe
c i e s . In such s i t u a t i o n s the apparent t o t a l molar s o l m

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

210

PRO-DRUGS

u b i l i t y , [ S ] , may be expressed as
T

[S]

= [S] + m [ S L ]
m

(eq. 3 ) .

When the concentration o f S i s at i t s s a t u r a t i o n value,


[ S ] , then eq. 3 becomes
Q

[S]

= [ S ] = m[S L ]
Q

(eq. 4 ) .

I t i s c l e a r from eq. 4 that i f S L i s a very s o l u b l e


species r e l a t i v e t o S then the apparent s o l u b i l i t y ,
[S]p may be g r e a t l y increased by complexation.
Another advantage of such a system, i m p l i c i t i n
eqs. 1 and 2 , i s that i concentrated s o l u t i o n
fS
and L, the e q u i l i b r i u
Such a s i t u a t i o n might be employed i n a concentrated
intravenous s o l u t i o n dosage form. When the concentrated s o l u t i o n i s d i l u t e d , as would occur upon the parent e r a l a d m i n i s t r a t i o n of the s o l u t i o n , the e q u i l i b r i u m
would be s h i f t e d t o the l e f t and r a p i d and extensive
d i s s o c i a t i o n of the complex would occur. Thus i f S was
a drug molecule whose s o l u b i l i t y was g r e a t l y increased
by complexation with L, a s o l u t i o n with a high concent r a t i o n of a s o l u b l e l i g a n d L would r e s u l t i n extensive
formation o f S L which would r e s u l t i n an increased
apparent s o l u b i l i t y of S. Such a s o l u t i o n would be
p h y s i c a l l y s t a b l e at e q u i l i b r i u m i n the dosage form but
would release the free drug from the complex upon d i l u t i o n i n blood without any required a i d from enzymes or
pH changes although the l a t t e r may a f f e c t the e q u i l i b rium i n some cases ( 2 3 . ) .
The e f f e c t s of g e n t i s i c a c i d on the s o l u b i l i t y of
hexamethylmalamine was studied at s e v e r a l pH values
(23.).
Phase diagrams o f the r e s u l t s at pH 3 . 5 , 4 . 0 ,
4 . 5 , and 5 are presented i n P i g . 3 I n a l l four cases
the apparent s o l u b i l i t y of hexamethylmalamine increased
as gentisate was added. At pH 3 - 5 and 4 . 0 , apparently
discontinuous curves were obtained and a s o l i d p r e c i p i t a t e was formed at added gentisate concentrations o f
greater than 0 . 8 M (at pH 3 . 5 ) and at 1 . 8 M or greater
(at pH 4 . 0 ) .
The reasons f o r the apparent and unexpected d i s c o n t i n u i t i e s were not f u l l y determined but could be due
i n part to some supersaturation and some very small
changes i n pH which occur upon p r e c i p i t a t i o n of the
complexes ( 2 3 . ) . I n the cases o f pH 4 . 5 and 5 - 0 , no
plateau regions corresponding t o those obtained at the
lower pH s were observed.
m

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

REPTA

Cytotoxic Agents in Parenteral Solutions

211

I t was apparent from the data i n F i g . 3 that s o l u t i o n s containing up to 5 mg of hexamethylmelamine per


ml could be prepared at pH 3 . 5 and 4 . 0 . Although i t
may have been p o s s i b l e to a t t a i n an even high apparent

Total Gentisate Species, M


Figure 3. Plot of the apparent aqueous solubility of hexamethylmalamine as a function of total added gentisate species
at 25 (23). Key:
pH 3.5;
pH 4.0;
pH
4.5;
pH 5.0. (The data points have been omitted for the
sake of clarity.)

s o l u b i l i t y f o r hexamethylmalamine at these pH v a l u e s ,
the abrupt d i s c o n t i n u i t i e s and the apparent s e n s i t i v i t y
of the observed s o l u b i l i t y to s l i g h t compositional

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

212

PRO-DRUGS

changes (as i n d i c a t e d i n P i g . 3 f o r s o l u t i o n s c o n t a i n ing hexamethylmalamine at concentrations exceeding 2 . 5


to 3 x 10""3 M) discouraged the use of such systems
which exceeded the p l a t e a u concentrations.
At the higher pH values o f 4 . 5 and 5 . 0 , p h y s i c a l l y
s t a b l e s o l u t i o n s c o n t a i n i n g concentrations of hexamethylmelamine exceeding 12 mg/ml may be prepared. A l though a greater t o t a l apparent s o l u b i l i t y o f hexamethylmelamine can be achieved at the higher pH values
shown, the concentration o f l i g a n d necessary to achieve
a given value increased sharply as the pH was i n creased. Thus i n choosing the s o l u t i o n f o r m u l a t i o n i t
i s necessary to consider the optimum composition w i t h
respect to concentration
f dru
d ligand
d th
pH. The system whic
through the N a t i o n a
pH 3 - 5 c o n t a i n i n g 5 mg of hexamethylmelamine and 6 mg
of g e n t i s i c a c i d per ml at pH 5 .
A more extensive complexation study than that r e ported here was made (23.) before the preceeding approach was adjudged t o be an acceptable s o l u t i o n t o the
problem of s o l u b i l i z i n g hexamethylmelamine. However,
the present d i s c u s s i o n does cover most of the s a l i e n t
aspects which l e d to the development and use o f the
complexed system as a r e v e r s i b l e drug d e l i v e r y system
s u i t a b l e f o r intravenous use.
Case I I I . Acetylacronycinium S a l t s and Complexes as a
Soluble S t a b i l i z e d Pro-Drug Form o f Acronycine.
Acronycine (VII) i s a c y t o t o x i c substance which
has e x h i b i t e d a c t i v i t y against a spectrum of tumor t e s t

OCH

OCH

HO

CH3
3
VII

VIII

Acronycine

Acronycinium Ion

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

REPTA

Cytotoxic Agents in Parenteral Solutions

213

systems ( 2 5 ) .
However, the low water s o l u b i l i t y (^2
mg/liter)T?6) has presented b i o a v a i l a b i l i t y problems
and t h i s has made the c l i n i c a l e v a l u a t i o n of t h i s agent
extremely d i f f i c u l t . The only p r e v i o u s l y reported a t tempt (27.) made to s o l u b i l i z e acronycine i n v o l v e d the
preparation of a c o p r e c i p i t a t e c o n t a i n i n g p o l y v i n y l
pyrrolidone (5 parts by weight) and acronycine (1 part
by weight). When the c o p r e c i p i t a t e was d i s s o l v e d i n
water, an apparent 5 f o l d increase i n the s o l u b i l i t y of
acronycine was obtained and the c o p r e c i p i t a t e demons t r a t e d enhanced antitumor a c t i v i t y when administered
i n t r a p e r i t o n e a l l y as a suspension. Whether or not the
enhanced a c t i v i t y was due to an increase i n the e q u i l i b r i u m s o l u b i l i t y of acronycin
(a
prob
ably the case) to a
c o p r e c i p i t a t e r e l a t i v e to acronycine alone was not
c l e a r . Nevertheless, because of the r a t h e r l a r g e a n t i cipated acronycine dose of 100 to 200 mg, and a d e s i r e
f o r that dose to be contained i n a volume of about 100
mis or l e s s ( 2 8 ) , the approach using the p o l y v i n y l p y r r o l i d o n e c o p r e c i p i t a t e apparently was not found s u i t able f o r intravenous use. Therefore i t was necessary
to pursue other means of s o l u b i l i z i n g acronycine.
The s o l u b i l i t y of acronycine i n a v a r i e t y of simple and mixed solvent systems was determined (29.) as
shown i n Table I I I . Prom such data i t was obvious that
Table I I I - Approximate Apparent S o l u b i l i t y of Acronycine i n Various Solvents at about 25 (29)
Solvent

S o l u b i l i t y (mg/ml)

chloroform
water
benzene
acetone
25% (v/v) acetone i n water
40$ (v/v) propylene g l y c o l i n
water

275
0.002
40
32
20
8

the s o l u b i l i t y problem was not one which could be


solved by using mixed solvent systems which would be
s u i t a b l e f o r intravenous purposes.
The s o l u b i l i t y data
together w i t h the r e l a t i v e l y non-polar s t r u c t u r a l f e a tures of the molecule and i t s r a t h e r inconspicuous
m e l t i n g behavior (m.p. 175-7 (26.)) suggested that the
low aqueous s o l u b i l i t y was p r i m a r i l y due to a l a c k of
h y d r o p h i l i c character r a t h e r than to strong i n t e r c r y s t a l l i n e i n t e r a c t i o n s . Therefore i t appeared, as i n

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

214

PRO-DRUGS

Case I I above, that the s o l u b i l i t y could most r e a d i l y


be increased by decreasing the a c t i v i t y of the d i s solved s o l u t e molecules. Since i t was d e s i r e d to i n crease the apparent s o l u b i l i t y of acronycine from a
value of about 2 mg per l i t e r up to 1 to 2 mg per ml,
which represents a 500 to 1000 f o l d i n c r e a s e , complexat i o n was r u l e d out due to the much smaller increases
normally obtained by that method.
The s o l u b i l i t y of acronycine i n a c i d i c s o l u t i o n i s
shown i n P i g . 4 and i t was obvious that i n order to ob-

t a i n the d e s i r e d s o l u b i l i t y the s o l u t i o n would have to


be at pH l e s s than zero, which i s i m p r a c t i c a l . However, the data i n F i g . 5 d i d demonstrate that through
the i n t r o d u c t i o n of a charge on the acronycine molecule

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

REPTA

Cytotoxic Agents in Parenteral Solutions

215

I t was p o s s i b l e to g r e a t l y increase apparent s o l u b i l i


t y . The problem then was one of i n t r o d u c i n g and main
t a i n i n g such a charge i n a medium compatible w i t h the
physiological state.

loool
600
400
2001-

CM

100
60
40

J3

20
10
6
4

6
PH

10

Figure 5. Plot showing the effects of pH on the observed


first order rate constant for the hydrolysis of the acetyhcronycinium ion at 25. The data points were obtained in a variety
of aqueous buffers at an ionic strength of unity (29, 30).

One method of a c h i e v i n g t h i s goal was through the


preparation of a d e r i v a t i v e which possessed e l e c t r o l y t e
p r o p e r t i e s . But, the major drawback i n t h i s case, as
i n Case I I , was the absence of chemical s i t e s which
would lend themselves to the formation of d e r i v a t i v e s
which would r e a d i l y and r a p i d l y release free acronycine
f o l l o w i n g intravenous a d m i n i s t r a t i o n .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

216

PRO-DRUGS

An i n s p e c t i o n of the acid-base chemistry of acronycine and r e l a t e d compounds (29.) u l t i m a t e l y provided


the s o l u t i o n to the problem of the l a c k of u s e f u l s i t e s
s u i t a b l e f o r preparing a chemically derived pro-drug.
I t appeared that the protonation of acronycine a c t u a l l y
occurj at the carbonyl oxygen which r e s u l t s i n aromatiz a t i o n of the n i t r o g e n - c o n t a i n i n g r i n g w i t h q u a t e r n a t i z a t i o n of the nitrogen atom as shown i n V I I I . This
c a t i o n i c species contains, i n a d d i t i o n to the p o s i t i v e
charge, a p h e n o l - l i k e group. Replacement of the a c i d i c
hydrogen atom with a l e s s l a b i l e s u b s t i t u e n t such as an
a l k y l or a c y l group was an apparent approach f o r maint a i n i n g an aromatic quaternary species s i m i l a r to V I I I .
For the reasons discussed i n Case I the a c y l d e r i v a
t i v e (IX) again seeme
derivative.
The f i r s t d e r i v a t i v e synthesized, a c e t y l a c r o n y c i nium (IXa) p e r c h l o r a t e , was prepared by heating acrony-

IX
Acylacronycinium

Ion

cinium perchlorate i n a c e t i c anhydride (2_9) The s a l t


obtained was found to hydrolyze r a t h e r r a p i d l y i n aqueous s o l u t i o n to y i e l d acronycine. Because of such hyd r o l y s i s only approximate values of the s o l u b i l i t y of
the s a l t could be obtained, but i t appeared that the
s o l u b i l i t y increase obtained was greater than 100 f o l d .
Since the apparent s o l u b i l i t y of the acetylacronycinium

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

REPTA

Cytotoxic Agents in Parenteral Solutions

217

i o n would be expected to be dependent upon the nature


of the anion i n the s a l t , a number of d i f f e r e n t s a l t s
were prepared and t h e i r approximate aqueous s o l u b i l i
t i e s are given i n Table IV. From t h i s data alone
i t appeared that phosphate or c h l o r i d e s a l t s might be
the most u s e f u l because of t h e i r high s o l u b i l i t i e s but
i t was found that a l l but the perchlorate s a l t were ex
tremely d i f f i c u l t to prepare and underwent r a t h e r r a p i d
decomposition i n the s o l i d s t a t e . Thus despite i t s
lower s o l u b i l i t y the acetylacronycinium perchlorate
s a l t was the substance which was chosen f o r more de
t a i l e d study ( 2 ) .
Table IV - Approximat Aqueou
Acetylacronycin

Solubilit

f Variou

(29).

acetylacronycine s a l t
perchlorate
sulfate
phosphate
bromide
chloride
a

a
s o l u b i l i t y (mg/ml)
^ .25
^ 1.5
^12
^ 6
^12

Concentrations are expressed i n terms o f acronycine


i n the s o l u t i o n .

The r e a c t i o n of acetylacronycinium p e r c h l o r a t e i n
water and aqueous b u f f e r was studied and i t was found
that the regeneration of acronycine from the a c e t y l a c
ronycinium i o n was q u a n t i t a t i v e ( 2_9) K i n e t i c studies
demonstrated that the rate of h y d r o l y s i s of the e s t e r
linkage was e s s e n t i a l l y independent of both pH and buf
f e r concentration over a range o f pH 0-7 However, at
pH greater than 8, hydroxide c a t a l y s i s was observed.
Some of the k i n e t i c data obtained (2) i s shown i n F i g .
5 where i t can be seen that at 25 the h y d r o l y t i c r a t e
at pH values below 8 i s about 2.8 1 0 " minute" which
corresponds to a h a l f - l i f e of about 25 minutes. From
studies of the temperature dependence of the h y d r o l y s i s
r e a c t i o n (29.), i t appeared that at a body temperature
of 37, the i n v i t r o h a l f - l i f e f o r h y d r o l y s i s would be
only about 5 minutes. Although such r a p i d h y d r o l y s i s
i s d e s i r e d f o l l o w i n g p a r e n t e r a l a d m i n i s t r a t i o n , the
o v e r a l l i n s t a b i l i t y presents problems i n the prepara
t i o n of the intravenous formulation. The d i f f i c u l t y
a r i s e s from a combination of f a c t o r s which include the
low s o l u b i l i t y of acronycine, the high apparent concen
t r a t i o n of acronycine (as the acetylacronycinium per2

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

218

PRO-DRUGS

c h l o r a t e ) d e s i r e d , and the r a t h e r r a p i d h y d r o l y s i s of
the acetylacronycinium i o n . As an i l l u s t r a t i o n of the
problem, consider a s o l u t i o n c o n t a i n i n g the a c e t y l a c r o nycinium perchlorate at a concentration corresponding
to the equivalent of 0.2 mg of acronycine per ml. Upon
h y d r o l y s i s of 5% of the acetylacronycinium s a l t , which
would occur i n only about 2 minutes (at room temperature) a f i v e f o l d supersaturated s o l u t i o n of acronycine
would r e s u l t . Moreover, the above example i s an o p t i m i s t i c one i n that no c o n s i d e r a t i o n was given to the
h y d r o l y s i s l i k e l y to occur during the preparation of
the s o l u t i o n .
In l i g h t of the above s t a b i l i t y problems, i t was
necessary to conside approache b which th h y d r o l y t
i c s t a b i l i t y of the
could be increased. In view of the p H - p r o f i l e ( F i g .
5), the s t a b i l i t y could not be enhanced by pH a d j u s t ment and thus more elaborate approaches had to be considered.
I n i t i a l l y , i t was f e l t that the i n t r o d u c t i o n of a
more bulky a c y l group would reduce the r a t e of hydrolys i s through s t e r i c e f f e c t s (15.). In order to evaluate
t h i s approach, the p e r c h l o r i d e s a l t s of IXb, IXc, and
IXd were prepared and the h y d r o l y s i s of each was studi e d at 25 and pH 7.0 or 7.5.
Somewhat s u r p r i s i n g l y ,
the r e s u l t s showed that the h y d r o l y t i c r a t e s f o r a l l
four of the acetylacronycinium perchlorate s a l t s were
e s s e n t i a l l y independent of the nature of the a c y l group
and the h a l f - l i f e f o r a l l was about 25 minutes. This
apparent complete l a c k of any dependence of the s t a b i l i t y on the s t r u c t u r e of the a c y l group prompted the
c o n s i d e r a t i o n of an a l t e r n a t i v e method f o r a t t a i n i n g
greater s t a b i l i t y .
E a r l i e r work by Higuchi and a s s o c i a t e (31332,33)
had shown that complex formation between organic spec i e s i n s o l u t i o n could a l t e r t h e i r chemical behavior
such as the s u s c e p t i b i l i t y to h y d r o l y s i s . Those r e s u l t s l e d to the c o n s i d e r a t i o n of complexing as a pot e n t i a l method f o r s t a b i l i z i n g the acetylacronycinium
ion.
For many of the same reasons mentioned i n Case I I ,
g e n t i s i c a c i d was chosen as a p o t e n t i a l l i g a n d and i t s
e f f e c t s on the s t a b i l i t y of acetylacronycinium i o n were
s t u d i e d . Some of the r e s u l t s obtained (29.) are shown
i n F i g . 6.
The observed decrease i n h y d r o l y t i c r a t e which occurs with i n c r e a s i n g gentisate concentration can be a t t r i b u t e d to the formation of complexes between the gent i s a t e and acetylacronycinium i o n s . Such complex spec i e s appear to decrease the s u s c e p t i b i l i t y of the acet-

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

REPTA

Cytotoxic Agents in Parenteral Solutions

219

ylacronycinium i o n to h y d r o l y s i s . Studies c a r r i e d out


i n more a c i d i c s o l u t i o n s showed a much l e s s e r enhancement of the acetylacronycinium i o n s t a b i l i t y by the
g e n t i s i c a c i d species, i n d i c a t i n g that the e f f e c t i v e
l i g a n d i s the gentisate i o n (29).

30H

. 0 8 O T W
.02
.04
.06
Gentisate Concentration [moles/liter]
Figure 6. Plot of the effects of the gentisate on the observed
first order rate constant for the hydrolysis of the acetylacronycinium ion at 25 (29, 30)

The non-linear data i n F i g . 2 has been shown (29.)


to f i t a model based on the formation of both 1:1 and
2:1 (gentisate i o n : acetylacronycinium ion) complexes.
In both such complexes the h y d r o l y t i c s t a b i l i t y of the
acetylacronycinium i o n was g r e a t l y enhanced r e l a t i v e to
the uncomplexed m a t e r i a l .
A f u l l and d e t a i l e d d i s c u s s i o n of the model used
i n the mathematical treatment of data such as that
shown i n F i g . 6 i s not e s s e n t i a l f o r the present d i s -

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

220

PRO-DRUGS

cussion and since such information i s a v a i l a b l e e l s e where (29.,30) 3 i t w i l l be omitted here.


From the data i n F i g . 6 i t may be shown that by
adding 0.2 M g e n t i s a t e i o n to the s o l u t i o n described
e a r l i e r ( c o n t a i n i n g the equivalent of 0.2 mg of acronycine per ml as the acetylacronycinium p e r c h l o r a t e s a l t )
h y d r o l y s i s of 5% of the acetylacronycinium i o n would
only occur a f t e r about 45-50 minutes. The production
of a saturated s o l u t i o n , w h i c h corresponds to 1% h y d r o l y s i s , would r e s u l t i n about 8-9 minutes. Although such
a system would s t i l l be a very marginal one f o r c l i n i c a l use due to i t s short-term s t a b i l i t y , i t represents
a s i g n i f i c a n t improvement over the system i n the absence of g e n t i s a t e
Continuin studie i n v o l v i n th
use of higher g e n t i s a t
of other more e f f e c t i v l i g a n d
g pursued
A l s o , recent r e s u l t s (34.) suggest that the s o l u b i l i t y
of f r e e acronycine i n aqueous g e n t i s a t e s o l u t i o n may be
enhanced s e v e r a l f o l d through complexation.
On the bas i s of the work done and that underway, i t i s f e l t that
i n the near future i t w i l l be p o s s i b l e to prepare a
pro-drug formulation of acronycine, as a r e c o n s t i t u t able s o l u t i o n , u t i l i z i n g a s o l u b l e acetylacronycinium
s a l t s t a b i l i z e d with a s u i t a b l e l i g a n d such as g e n t i s ate. A good deal more work remains to be done, both i n
v i t r o and i n v i v o , before that goal can be achieved.
However, regardless of the f i n a l outcome of t h i s study
i n terms of useable product, i t has been demonstrated
that r a t h e r s o p h i s t i c a t e d and complex systems may be
developed and used as a pro-drug approach to overcoming
problems i n the formulation of intravenous s o l u t i o n s of
c y t o t o x i c agents.
Summary
The above cases represent some of the more succ e s s f u l a p p l i c a t i o n s of the pro-drug approach to overcoming problems encountered i n the intravenous d e l i v e r y
of c y t o t o x i c agents.
I t should be obvious that the development of s u i t a b l e pro-drugs i n v o l v e s a good deal
more than simply preparing some new chemical species.
There must be a r a t i o n a l e f o r choosing the new species
and the t o t a l system must be r e v e r s i b l e under b i o l o g i c a l c o n d i t i o n s i n order to achieve any degree of success. Whether or not any of the pro-drug systems d i s cussed ever becomes medically important i s l a r g e l y dependent upon the b i o l o g i c a l p r o p e r t i e s inherent i n the
parent drug i t s e l f . Therefore, while the approaches
i l l u s t r a t e d here i n the cases of c y t o t o x i c agents may
not prove to be important i n these p a r t i c u l a r systems

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5. REPTA

221
Cytotoxic Agents in Parenteral Solutions

they may ultimately be extrapolated to other systems


involving other promising drugs.
It has been this author's objective in the above
cases to discuss and review the rationale which was in
volved in the development and evaluation of the ap
proaches used and not to delve into the details of
these studies which are or will be reported elsewhere
as indicated in the literature citations and especially
references (8)(23)(29) and (30).
Acknowledgements
The author wishes to recognize the partial support
of this work by contract #N01-CM-23217 from the Divi
sion of Cancer Treatment
National Institute of Health, Department of Health, Ed
ucation and Welfare. In addition, special thanks are
extended to those persons who were responsible for much
of the work which has been reviewed here. A l i s t of
these persons includes D. W. A. Bourne, T. Higuchi,
C-H. Huang, B. Krielgard, B. J . Rawson, R. D. Shaffer,
. B. Sloan and W. Waugh.
Literature Cited
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.

Benet, L. Z., in "Drug Design", E. J . Ariens, ed.,


Vol. IV, pp. 9-11, Academic Press, New York, 1973.
Wagner, J . G., "Biopharmaceutics and Relevant
Pharmacokinetics", pp. 61-2, Drug Intelligence
Publications, Hamilton, Ill., 1971.
Hess, B . , "Enzymes in Blood Plasma", pp. 7-18,
Academic Press, New York, 1963.
Beck, M. T., "Chemistry of Complex Equilibria",
pp. 21-3, Van Nostrand Reinhold Co., London, 1970.
Medical News, JAMA (1974), 230, 189.
LePage, G. ., Advances in Enzyme Regulation
(1970), 8, 323.
Cohen, S. S., Prog. in Nucleic Acid Res. Mol.
Biol. (1966), 5, 1.
Repta, A. J., Rawson, B. J., Shaffer, R. D.,
Sloan, . B . , Bodor, ., and Higuchi, T., J .
Pharm. Sci. (1975), 64, 392.
Davignon, J., and Cradock, J . (of the National
Cancer Institute) personal communications, Aug.,
1972.
Bloch, ., Robins, M. J., and McCarthy, J . R.,
J r . , J . Med. Chem. (1967), 10, 908.
Koshima, R., and LePage, G. ., Cancer Res. (1968),
28, 1014.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

222

PROD
- RUGS

12. White, F. (of the National Cancer Institute, per


sonal communication, May, 1973.
13. Wheeler, L. M. (of Parke, Davis and Co.) personal
communication, Aug., 1972.
14. Rawson, B. J., Unpublished data (from this labora
tory).
15. March, J., "Advanced Organic Chemistry; Reactions,
Mechanisms and Structures", pp. 233-245, McGrawHill,
New York, 1968.
16. Blum, R. H., Livingston, R. B . , and Carter, S. K.,
Europ. J . Cancer (1973), 9, 195.
17. Takita, H. and Didolkar, M. S., Cancer Chemother.
Rep. (1974), 58, 371.
18. Cradock, J . (of the National Cancer Institute)
personal communication
19. National Cancer Institute, Progress reports from
the Warner-Lambert Research Institute (Contract
#72-3250), Aug., 1972.
20. "Toxic Substances List", p. 472 (1973), U. S. De
partment of Health, Education and Welfare, Rock
ville, Maryland, 1973.
21. "Merck Index", 85th Ed., p. 486, P. G. Stecher,
ed., Merck and Co., Rahway, New Jersey, 1968.
22. Higuchi, T. and Pitman, I. H., J . Pharm. Sci.
(1973), 62, 55.
23. Krielgard, B., Higuchi, T., and Repta, A. J.,
(Manuscript submitted to J . Pharm. S c i . , January,
1975).
24. Higuchi, T. and Connors, . ., "Advances in Ana
lytical Chemistry and Instrumentation", Vol. IV,
pp. 128-193, C. N. Reilly, ed., Interscience, New
York, 1965.
25. Svoboda, G. H., Poore, G. ., Simpson, P. J., and
Bodor, G. B., J. Pharm. Sci. (1966), 55, 758.
26. C. A. Hewitt, Stanford Research Institute reports
to the National Cancer Institute, Nos. 758 and 770
1968.
27. Svoboda, G. H., Sweeney, M. J., and Walking, W.
D., J . Pharm. Sci. (1971), 60, 333.
28. Davignon, J. P. (of the National Cancer Institute)
personal communication, March, 1972.
29. Bourne, D. W. ., Doctoral Dissertation, Univer
sity of Kansas, 1974.
30. Bourne, D. W. ., Higuchi, T., and Repta, A. J.,
A presentation to the Basic Pharmaceutical Section
of the Academy of Pharmaceutical Sciences, F a l l
meeting, 1973, San Diego. Detailed manuscripts on
the work have been submitted for publication to J .
Pharm. Sci. and may be expected to appear in print
in late 1975.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

5.

31.
32.
33.
34.

REPTA

Cytotoxic Agents in Parenteral Solutions

223

Higuchi, T. and Lachman, L., J. Pharm. S c i . (1955)


44, 5 2 1 .
Lachman, L. and Higuchi, T., i b i d . ( 1 9 5 7 ) , 46, 3 2 .
Lachman, L., Ravin, L., and Higuchi, T., i b i d .
(1956), 4 5 , 290.
Huang, C.-H., Master's D i s s e r t a t i o n , University of
Kansas, 1 9 7 5 .

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

6
The Effect of a Pro-drug of Epinephrine (Dipivalyl
Epinephrine) in GlaucomaGeneral Pharmacology,
Toxicology, and Clinical Experience
DAVID A. McCLURE
Alergan Pharmaceuticals, Irvine, Calif. 92664
Epinephrine ha
treatment of the conditio
is a disease where the pressure within the eye increases to a point where damage to the visual apparatus occurs and, if left unchecked, could lead to
a significant deminuation in visual acuity and eventually, blindness.
The reasons for this pressure increase are
numerous and we will touch on these shortly. However,
first, let us examine the anatomy of the eye and how
glaucoma comes about. Figure 1 shows a cross section
of an eye. From the crystalline lens backward is one
chamber filled with a thick, viscus fluid that does
not move. Another chamber, that is in front of the
lens and the Zonule ligament, is actually composed of
two chambers with fluid circulating between. This
fluid is called the aqueous humor. The path of the
aqueous humor is from the ciliary body, through the
pupil, and out the trabecular meshwork and an area
called the Canal of Schlemm. Figure 2 is a close-up
of the area of fluid circulation and drainage.
Now, we mentioned earlier that glaucoma results
from an increase in the intraocular pressure (IOP).
The normal IOP averages around 17 mmHg. This pressure
can increase in 3 ways: 1) a greater influx of fluid
into the envelope of the eye while maintaining a constant outflow, 2) a decreased outflow while maintaining a constant inflow or, 3) a combination of the
two. We are concerned primarily (in simple primary
glaucoma) with a decrease in the outflow of aqueous
fluid. Now what happens if the IOP goes up 10 to 20
mmHg from normal and is maintained at that level? In

224
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

6.

MC CLURE

Pro-drug of Epinephrine in Glaucoma

225

o t h e r words, why i s glaucoma bad?


The p r e s s u r e t h a t
i s b u i l t up i n the f r o n t p a r t o f the eye i s r e f l e c t e d
t o the back p a r t o f the eye where, o f c o u r s e , the
most v i t a l p a r t s o f the eye r e s i d e .
Figure 3 i s a
photograph o f the r e t i n a .
T h i s i s the p o i n t where
the o p t i c nerve and b l o o d v e s s e l s emerge.
As the
p r e s s u r e i n c r e a s e s , the d i s c c o n t a i n i n g the nerve
and b l o o d v e s s e l s i s pushed back and a c o n d i t i o n
c a l l e d cupping occurs.
This r e s u l t s i n a decreased
v i s u a l f i e l d ( p e r i p h e r a l ) and as the c o n d i t i o n c o n t i n u e s , v i s i o n gets worse.
C h r o n i c s i m p l e glaucoma has been t r e a t e d b o t h
m e d i c a l l y and s u r g i c a l l y .
M e d i c a l l y , t h e r e are a
v a r i e t y o f drugs c u r r e n t l y a v a i l a b l e .
These a r e
primarily cholinergi
a d r e n e r g i c a g e n t s such as e p i n e p h r i n e .
There a r e
p r o s and cons f o r the use o f each g r o u p .
F o r example,
the c h o l i n e r g i c s cause m i o s i s ( i . e . , a s m a l l p u p i l )
and enough accomodative spasm ( i . e . , d e c r e a s e d a b i l i t y
to focus) t h a t r e a d i n g i s d i f f i c u l t , d r i v i n g at n i g h t
is d i f f i c u l t , etc.
In a d d i t i o n , because some c h o l i n e r g i c s do n o t p e n e t r a t e i n t o the eye v e r y w e l l , the
f r e q u e n c y o f a d m i n i s t r a t i o n i s a t an i n c o n v e n i e n t ,
6 t o 8 times p e r d a y .
In a d d i t i o n , because most o f
the c h o l i n e r g i c s are e s t e r s o r l a c t o n e s , the h i g h
l e v e l o f l o c a l e s t e r a s e enzymes r e q u i r e s the i n c r e a s e d
frequency of a d m i n i s t r a t i o n .
Even w i t h a l l t h e s e
i n c o n v e n i e n c e s , m i o t i c s are used e x t e n s i v e l y and t e n d
t o work q u i t e w e l l .
T h e i r mechanism o f a c t i o n i s
p r o b a b l y v a s o d i l a t i o n which p e r m i t s a g r e a t e r e x i t
o f aqueous f l u i d from Schlemm's C a n a l .
The main a d r e n e r g i c compound used today i s
epinephrine.
T h i s compound appears t o have a t w o f o l d
e f f e c t on the maintenance o f IOP.
The f i r s t i s the
b e t a a d r e n e r g i c mechanism w h i c h , s i m i l a r t o the
c h o l i n e r g i c a g e n t s , opens up the e p i s c l e r a l b l o o d
v e s s e l s p e r m i t t i n g a g r e a t e r f l u i d l o s s from the
c a n a l o f Schlemm.
T h i s appears t o be b r o u g h t about
by the s t i m u l a t i o n o f e p i n e p h r i n e on the b e t a a d r e n e r g i c r e c e p t o r s i n e p i s c l e r a l b l o o d v e s s e l s and a l s o
t h o s e r e s i d i n g i n the C a n a l o f Schlemm and perhaps
even i n the t r a b e c u l a r meshwork.
In a d d i t i o n ,
e p i n e p h r i n e has a v a s o c o n s t r i c t i v e mechanism, i . e . ,
the a l p h a a d r e n e r g i c mechanism, which may a c t by
d e c r e a s i n g the p r o d u c t i o n o f aqueous f l u i d from the
c i l i a r y body.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

226

PRO-DRUGS

Figure 1

Figure 2

Figure 3
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

6.

MCCLURE

Pro-drug of Epinephrine in Glaucoma

227

Even though e p i n e p h r i n e appears t o have t h i s


unique t w o f o l d mechanism o f a c t i o n , a number o f
problems a r i s e , l i k e the c h o l i n e r g i c s , w i t h i t s u s e .
T a b l e 1 i l l u s t r a t e s some o f t h e s e p r o b l e m s .

Table 1
Problems o f E p i n e p h r i n e use i n Glaucoma
I.
II.

III.
IV.

Duration of Action
Side E f f e c t
A.
Ocular
B.
Systemic
Bioavailability
Stability

Duration of a c t i o n .
When a d m i n i s t e r e d i n t r a v e n o u s l y , the c a r d i o v a s c u l a r , pulmonary and o t h e r
e f f e c t s from e p i n e p h r i n e are o v e r w i t h i n 10 m i n u t e s .
When i n s t i l l e d i n t o the e y e , a d i l a t e d p u p i l o c c u r s
i n 10 t o 15 minutes and the d i l a t i o n l a s t s up t o an
hour.
The d u r a t i o n o f l o w e r i n g o f IOP from e p i n e p h r i n e can be seen i n F i g u r e 4.
The c o n c e n t r a t i o n o f
e p i n e p h r i n e (2%) used i n t h e s e d a t a r e p r e s e n t s the
highest concentration a v a i l a b l e .
It i s presented
h e r e t o demonstrate the peak a c t i v i t y time o f 4 hours
and a d u r a t i o n o f a c t i o n o f between 12 and 24 h o u r s .
The m e t a b o l i c pathways f o r e p i n e p h r i n e can be seen
i n F i g u r e 5.
The s i d e e f f e c t s o c c u r r i n g from e p i n e p h r i n e
t o p i c a l l y a p p l i e d t o the eye are b o t h l o c a l o c u l a r
s i d e e f f e c t s and s y s t e m i c .
T a b l e 2 i l l u s t r a t e s some
of these e f f e c t s .
To g e t around a l l t h e s e problems w i t h e p i n e p h r i n e , we were f o r t u n a t e t o get a chance t o examine
an analogue o f e p i n e p h r i n e and p o s s i b l y what i s f e l t
t o be a p r o - d r u g o f e p i n e p h r i n e .
F i g u r e 6 shows the
c h e m i c a l s t r u c t u r e o f e p i n e p h r i n e and i t s p r o - d r u g ,
d i p i v a l y l e p i n e p h r i n e (DPE).

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

228

PRO-DRUGS

40

30

-A

20
AVERAGE OF
44 UNTREATED
GLAUCOMATOUS
EYES

10

1
TIME (HOURS)
Archives of Ophthalmology

Figure 4. Response to 1 drop 2% epinephrine (1)

Figure 5
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

6. MCCLURE

Pro-drug of Epinephrine in Glaucoma

Table 2
Ocular Side E f f e c t s

of T o p i c a l Epinephrine

I . Hyperemia
II. Mydriasis
-r-r-r

V . Browache
(Photophobia)

I I I . C o r n e a l Edema
_ _..
.
_
.
IV. A l l e r g i c S e n s i t i v i t y
Systemic
I.

V I . Adrenochrome
DepOSltS

. -,

Side E f f e c t s

V I I . Tolerance
VIII.

Maculaopathy

of T o p i c a l Epinephrine

Cardiovascula
A . Cardiac Arrhythmias
B. Blood Pressure E l e v a t i o n
C. Cerebrovascular

II. Pallor,

Dizziness,

III. Fear, Anxiety,

Accidents
Tremor

Tenseness,

Restlessness

CH-CH -N
2

CH,

EPINEPHRINE (Mol.Wt. 183.20)


CH-CH-.N'
^CH,

DIPIVALYL EPINEPHRINE (DPE,Mol.Wt. 587. 904)

Figure 6

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

229

230

PRO-DRUGS

Table 3
Proposed Advantages o f DPE
I.
II.

Increased Duration o f A c t i o n
Increased B i o a v a i l a b i l i t y

III.

Increased

Potency

IV.

Decreased

Side

V.

Increased

Stability

Effects

T a b l e 3 shows t h e p r o p o s e d advantages o f DPE


epinephrine.
Why an i n c r e a s e d d u r a t i o n o f a c t i o n w i t h DPE?
F i g u r e 7 i l l u s t r a t e s t h e p o s s i b l e r e a s o n f o r an i n creased duration of a c t i o n .
The main m e t a b o l i c p a t h way o f e p i n e p h r i n e i s v i a an enzyme c a l l e d c a t e c h o l o - m e t h y l t r a n s f e r a s e (COMT).
T h i s enzyme m e t h y l a t e s
the meta h y d r o x y l i n e p i n e p h r i n e .
T h i s h y d r o x y l , as
w e l l as t h e p a r a h y d r o x y l a r e n o t f r e e i n DPE. The
p i v a l y l m o i t i s a r e p r o b a b l y s l o w l y removed by l o c a l
e s t e r a s e enzymes so t h a t COMT c a n then a c t .
This
would t a k e a p r o l o n g e d p e r i o d o f t i m e , thus c a u s i n g
a "sustained release" epinephrine.
over

ON
N CO

CHCHgNHCH

NO
METANEPHRINE

CHCN NHCH
2

NO

3.4 Dl HYDROXY
MANDELIC
ALDEHYDE

Figure 7
In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

6. MCCLURE

Pro-drug of Epinephrine in Glaucoma

231

Why an i n c r e a s e d b i o a v a i l a b i l i t y ? I t i s v e r y
e v i d e n t t h a t DPE i s much more l i p o p h i l i c than e p i n e p h r i n e by v i r t u e o f t h e two l a r g e p i v a l y l groups a t t a c h e d t o t h e two h y d r o x y l s o f e p i n e p h r i n e .
In a d d i t i o n , DPE m a i n t a i n s a h i g h degree o f h y d r o p h i l i c i t y .
T h e r e f o r e , by i t s d u a l s o l u b i l i t i e s , i t f i t s i n v e r y
n i c e l y t o t h e p r e s e n t day a b s o r p t i o n t h e o r i e s . The
c o r n e a o f t h e eye i s t h e b a r r i e r drugs must overcome
i n o r d e r t o be absorbed i n t o t h e e y e . The c o r n e a i s
composed o f t h r e e l a y e r s :
an e p i t h e l i u m and an e n d o t h e l i u m , b o t h o f w h i c h r e q u i r e drugs t o be l i p o p h i l i c
i f t h e y a r e t o be absorbed a n d , t h e s t r o m a ; s a n d wiched between t h e e p i t h e l i u m and e n d o t h e l i u m t h a t
r e q u i r e d a d r u g t o be h y d r o p h i l i c f o r p e n e t r a b i l i t y .
By t h e d u a l s o l u b i l i t
the eye i s g r e a t e r t h a n t h e l e s s l i p o p h i l i c e p i n e phrine molecule.

20,

TIME IN HOURS

Figure 8. Comparison of IOP effects in rabbits

Why an i n c r e a s e d potency?
I f more m a t e r i a l g a i n s
a c c e s s t o t h e i n s i d e o f t h e eye i t w i l l t h e n , on a
r e l a t i v e b a s i s , be more p o t e n t .
F i g u r e 8 shows a
comparison between 0.5% e p i n e p h r i n e and 0.16% DPE
DPE on t h e IOP o f u n a n e s t h e t i z e d r a b b i t s .
The e f f e c t s

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

232

PRO-DRUGS

are o b v i o u s . F i g u r e 9 i l l u s t r a t e s a dose-response o f
DPE on c a u s i n g p u p i l l a r y d i l a t i o n .
On t h e d i f f e r e n t i a l s i d e e f f e c t s t u d i e s , a
comparison o f DPE t o e p i n e p h r i n e r e l a t i v e t o b l o o d

bJ 01

If

TIME-(HOURS)

j
< kl

Figure 9. Mean percent differences in mydriatic response between


treated eyes and control eyes when treated with DPE: 0.20% (O),
0.25% (),0.30% (A),0.40% (O),I.O0% ()

p r e s s u r e and h e a r t r a t e e f f e c t s a f t e r i n t r a v e n o u s
a d m i n i s t r a t i o n was c a r r i e d o u t i n dogs and c a t s .
F i g u r e s 10 and 11 compare t h e e f f e c t s o f i n t r a
v e n o u s l y a d m i n i s t e r e d DPE and e p i n e p h r i n e on t h e
b l o o d p r e s s u r e and h e a r t r a t e i n a n e s t h e t i z e d d o g s .
I t i s e v i d e n t t h a t DPE has s i g n i f i c a n t l y l e s s e f f e c t
on b l o o d p r e s s u r e and h e a r t r a t e t h a n e p i n e p h r i n e .
In a s i m i l a r f a s h i o n , t h e e f f e c t o f DPE and e p i n e
p h r i n e on t h e b l o o d p r e s s u r e o f a n e s t h e t i z e d a t r o p i n i z e d c a t s may be seen i n F i g u r e 12.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

MCCLURE

Pro-drug of Epinephrine in Glaucoma

TIME (MINUTES)

Figure 10. feet of I.V. epinephrine and DPE on blood pressure


in dogs

Figure 11. Effect of I.V. epinephrine and DPE on heart rate in dogs

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

234

PRO-DRUGS

DPE i s , t h e n , about 100 t o 400 times weaker


than e p i n e p h r i n e i n a f f e c t i n g the c a r d i o v a s c u l a r
systems o f dogs and c a t s .
I t i s about 100 times
more p o t e n t t h a n e p i n e p h r i n e i n i t s a b i l i t y t o lower
IOP.

DOSE (XIO-SMOLES/KC)
Figure 12. The effect of epinephrine and DPE on the arterial blood
pressure of the anesthetized atropinized cat

F i n a l l y , t h e e f f e c t o f DPE on humans w i t h
glaucoma may be seen i n F i g u r e 13.
I t c a n be seen
t h a t t h e response e l i c i t e d by DPE i s p r o n o u n c e d .
If
one s u b t r a c t s t h e c o n t r a l a t e r a l c o n t r o l eye (normal
d i u r n a l response) from t h e t r e a t e d e y e , i t c a n be
see t h a t DPE produced a marked r e d u c t i o n i n IOP.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

6. MCCLURE

Pro-drug of Epinephrine in Glaucoma


235

Summary
The dipivalyl analogue of epinephrine has been
shown to produce fewer side effects and greater
potency than the parent compound. These results have
been found in both animal and human studies. More
studies are to be carried out which will amplify
these data and perhaps lead to a better understanding
of the mechanism of action of this epinephrine prodrug.
Literature Cited
1. L. L. Garner, W. W. Johnstone, E. J.
Ballintine, M. E. Carroll, Arch Opth. (1959), 62, 230.

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

INDEX
A
Absorption barriers
9
Absorption of drugs
4,196
Accidental pro-drugs
90
Acetamidopenicillins
131
Acetaminophen
81,90
O-acyl and O-carbonate esters of ....
81
2- ( p- Aeetaminophenoxy ) tetrahydropyran
81
Acetonide formation .,
38
a-Acetoxyethyl phenylmalonyl
chloride
126
Acetoxymethyl-7-aminocephalosporanate
13
Aeetoxymethyl-7- ( D-a-aminophenylacetamido)cephalosporanate
134
Acetoxymethyl-7- ( D-a-azidophenylacetamido)cephalosporanate
134
Acetoxymethyl-7- ( 2'-thienylacetamido) cephalosporanate
134
Acetylacronycine salts
217
Acetylacronycinium
complexes
212
ion
215,219
perchlorate
216
salts
212
4"'-Acetyldigoxin
40
3- Acetyl-5,5-diphenylhydantoin
178
Acetylsalicylic acid
76
Acidic media
126
Acronycine, pro-drug of
212
Acronycine, solubility of
213,214
Acronycinium ion
212,216
7-Acylacephalosporanic acid
138
7-Acylaminocephalosporanic acid 138,139
7-Acylaminodesacetoxy cephalosporanic acid
138
Acylanid
41
Adamantoyl esters
26
Addison's disease
47,55
Adenine arabinoside, 5' phosphate of
71
Adenosine
200,202
3',5'-Adenosine monophosphate
25
Adrenergic agents
225
Adrenergic mechanism
225
Adriamycin-14-octanoate
75,91
Aglycone
2
Alcoholic patients
19
Allopurinol
41
Amines
15
Amino acid carbamates
..
66

-Aminobenzyl penicillin
118,130,131
7-a-Amino cyclohexadienylalkyl
cephalosporanic acid
139
-Amino cyclohexadienylalkyl
penicillin
130
6-Aminopenicillanic acid
122,125
p-Aminosalicylic acid
49
Ammonium drugs, quaternary
26
Amodiaquine
59
Amphotericin
79
methyl ester
79
Ampicillin
30, 118, 122-124
deoxylincomyci
,
Antagonist, narcotic
62
Anthracene glycosides
2
Antibacterial activity
143
Antibiotics
^-lactam
118
macromolecular salts of
33
oral bioavailability of
30
polar
33
pro-drug approach to
116
Anticonvulsant
74,154,184
Anti-inflammatory agents,
nonsteroidal
76
Antifungal agent
42
Antihypercholesteremic agent
78
Antimicrobial agents, sulfonamide ...
90
Aqueous solubility ( See also Water
soluble)
40,66,73,155
Ara-A
200-202,205
-5' formate
201,202,205,206
Ara-H
201
l-/?-D-Arabinofuranosyladenine,
pro-drug of
200
a-Aryl-0-aminoethyIpenicillin
130
Ascorbic acid
22,36,88
3-phosphoryl ester of
36
Aspirin
1,76
6-Azauridine
23

Barrier(s)
blood brain
of drug to taget organ
physical, chemical, and social
Behavior, psychotic
Benzathine
penicillin
Benzoic acid derivatives

239

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

85
6
7
55
3
3,86
76

240

PRO-DRUGS

Benzophenone
185
Bioactivities
145, 147
Bioavailability
of antibiotics, oral
30
dose-dependent
44
of a drug
196
of steroids
45
Bioreversible
chemical modification of drugs
73
derivatives
1
pro-drugs, water soluble
175
Biotransformation
2
Birth control
5
,'-Bis ( 2-hydroxycyclohexyl ) -ethylenediamine-,' diacetic acid .... 35
Bis-trichloroethyl carbonate
82
Blood
ara-A-5' formate and ara-A i
brain barrier (BBB)
9,85
levels, thiamine
19,20
pressure of cats
234
pressure of dogs
233
of rats, bioactivities in
147
thiamine levels in
21
Brain dopamine
9
Buffers
187, 188
Burn therapy
36
N -Butanoyl derivatives
25
6

Calcium 4-benzamidosalicylate
49
Cancer chemotherapeutic agent
200
N-Carbamoyl-2,2-diphenylglycine,2( diethylamino ) ethyl ester,
hemisulfate
184,186
Carbenicillin
125,131
esters of
126,128
disodium salt
126
indanyl sodium-sodium 6-[2phenyl-2- ( 5-indanyloxycarbonyl)] acetamido peni
cillinate
125
N-Carbethoxychlorphentermine
15
N-Carbethoxy-2,2-diphenylglycine .... 160
3-Carbethoxy-5,5-diphenylhydantoin
156,178
N- ( Carbethoxymethyl ) -3-acetoxymethyl-7- ( 2'-thienylacetamido ) 2-cephem-4-carboxylic acid
amide
132
IV- [ 1-Carbo-t-butoxy ) ethyl] -7- ( 2'thienylacetamido ) cephalosporanic acid amide
132
-Carboxybenzylpenicillin
125
Carcinoma
55,69,83
Cat
234
Catecholamines
9,14,47
Catechol-0-methyl transferase
230
Central nervous system (CNS)
9,15,29
Cephaloglycin
135,140

Cephalosporin
128
C amides
132
C esters
128
C esters
132
GI absorption of
134
pro-drugs
135,138
Cerebrospinalfluidconcentration
20
Chelating agents, heavy metal
34
Chemotherapeutic agent, cancer
200
Chloral hydrate
81
Chloramphenicol
68,71,80
succinate
68, 71
Chloroguanide
59,60
p-Chlorophenoxyisobutyric acid
78
ethyl ester
78
Cholinergic agents
225
4

alkyl esters
141
bioactivity concentrations, serum ... 145
-2,3-biscarbonate ester
142
[7( S )-chloro-7-deoxylincomycin]
hydrochloride
137
-2-cyanoethyl phosphate
146
2,3-diesters
143
mean serum concentrations of . .148,149
-3-monocarbonate ester
142
-2-monoester
142,143
-2-phosphate
144,146,148,149
subcutaneous dose of
147
Codeine
2
Complex of a drug
3
Complexation
209
Corneal absorption
35
Corticosteroids
38
Covalent bond linkage
3
Covalent chemical modification
86
Crystal lattice energies
42
6-N'-Cyanoamidopenicillin
130
Cyanoethyl phosphate ester
intermediate
146
Cycloguanil
59,60
Cytosine arabinoside, 5'acyl
derivatives of
51
Cytotoxic agents
196
D

Darvon
Deanol
Degradations, solid state
Delivery
of polar anticiotics
problems of cytotoxic agents,
intravenous
site specific
Derivative
formation
in vivo liability of a drug

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

86
90
88
33
196
83
33
117

241

INDEX

Derivative (continued)
nontoxic
117
over the parent molecule,
advantage of
117
scale-up of
117
stability of
117
synthesis and purification of
116
Desacetylrifampicin
137
Desoxycorticosterone
47
trimethylacetate
55
Dexamethasone-21-phosphate
67
4,4'-Diaeetoamidodiphenylsulfone .... 59
,-Diacetyldopamine
13
3,6-Diacetylmorphine
48
0,S-Diacetylthiamine
16
4,4'-Diaminodiphenylsulfone
50,59
,-Dibenzoylthiamine disulfide
16
N -2'-0-Dibutanoyladenosine-3',5'
monophosphate, diburanoy
derivative
25
Dichloralphenazone
81
O-S-Diethoxycarbonylthiamine
16
Dilantin
184
^-N,N-Diethylaminoethyl-2,2diphenylglyoinate
161
^-iV,iV'-Diethylaminoethyl-2,2diphenylhydantoate
162,164
^-N',N-Diethylaminoethyl-2-ethyl-5methyl-2-phenylhydantoate
157
^N',N'-Diethylaminoethyl-5-methyl2,2-ethylphenylhydantoate
72
Diethyldithiolisophthalate
89
Diethylenetriaminepentaacetic acid .. 34
Diethylstilbesterol
69, 83
4,4'-Diformamidodiphenylsulfone
50
Digoxin
40
Dihydroxy groups in some steroids .... 38
L-3,4-Dihydroxyphenylalanine
10
N-N-Dimethylaminoethanol
90
Dimethylol urea
64
6- ( 2,2-Dimethyl-5-oxo-4-phenyl-limidazolidinyl) penicillanic acid 119
2,2-Diphenylglycine
159,171
5,5-Diphenylhydantoin (DPH)
pro-drug
154,175,184
aqueous and lipid solubility of
155
bioavailability
154
conversion of pro-DPH
to
165,169,185,187,188
in dogs
190,191,193
in rats
187-189, 191-193
separation of pro-DPH and
186
Diphenylmethane derivatives
185
4,4-Diphenyl-2,5-oxazolidinedione ... 160
Dipivalyl derivative
35
Dipivalyl epinephrine
(DPE)
227, 230-234
Distribution of drugs
4
0,0-Di(trimethylsilyl) dopamine
13
Dogjs)
effect of I.V. epinephrine and DPE

Dogs (continued)
on blood pressure and heart
rate of
233
plasma levels at DPH and
pro-DPH in
190, 191
tissue levels of pro-DPH and
DPH in
193
urine, indanol metabolites in
129
L-Dopa
10,44
esters
11
Dopamine
12,15
Dose dependent bioavailability
44
Dose-response
231
Drug(s)
absorption or bioavailability of
196
absorption, distribution, metabo
lism, and excretion of
4

derivative, in vivo lability of a


117
formulation problems of
85
-induced gastric ulceration
75
metabolite
6
patient acceptance of
80
polar
35
quaternary ammonium
26
-receptor interaction
4
solubility of (See also Aqueous
solubility and Water soluble)
9, 40, 60,66, 73, 81
stabilization of
43,85
to target organ, barriers of
6
taste problem of
80,81
time profile of a
4
toxicity of
73,75

Epinephrine ...12, 35, 86, 224-229, 233, 234


Epinephryl borate
87
Erythromycin
31, 32, 86
Ester formation
59
Estradiol
47,55,83
3-benzoate-17-cyclooctenyl ether ... 55
l-Ethoxyethyl-4-allopurinyl ether
41
Ethylchloroformate
159
Ethylenediaminetetraacetic acid
34
Ethyl mercaptan
89
5-Ethyl-3-methyl-5-phenylhydantoin .. 154
5,5-Ethylphenylhydantoin
74
5-Ethyl-5-phenylhydantoin
180
Excretion of drugs
4
Extraction scheme
186
Eye
226
F
First order rate constant
206
First pass effect
9,44
Fluid circulation and drainage in the
eye
26
Fluids, physiological
60

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

242

PRO-DRUGS

Fluoinolone
acetonide-21-acetate
Fluphenazine decanoate
Fluphenazine dihydrochloride
Fluphenazine enanthate
-Fluphenthixol dihydrochloride
Formaldehyde
Formulation problems of drugs
9- ( 5-O-Formyl-0-D-arabinofuranosyl) adenine drug
3-Formyl rifamycin SV

37
38
55
55
55
57
1,64
85
200
137

G
Gastric irritants
76
Gastric ulceration, drug-induced
75
Gastrointestinal (GI) tract
9,134
Gentisate complex
3
Gentisic acid
207,209,21
Glaucoma
35,224,227,23
Griseofulvin
42
-4'-alcohol
43
-4'-carboxymethoxime
43
-4'-hemisuccinate
43
-4'-oxime
43
H
-Haloalkylamines
28
Heart rate in dogs
233
Heavy metal chelating agents
34
p-Heptyloxyhydrocinnamyltestosterone
54
Heroin
48
Hetacillin
119,122,123,131
Hexamethylmelamine
complexation of
209
-gentisic acid complexes
3,207
pro-drug of
3, 207
solubility of
208,210,211
3, Hexoxycarbonyl-5,-diphenylhydantoin
179
Human serum levels
24
Hydantoic acid
158,176
Hydantoins
176,180
Hydrocortisone-21-phosphate
67
Hydrolysis
43, 120, 121
Hydroxydione
71
2-Hydroxyethylenediamine-N,N,IVtriacetic acid
34
N-Hydroxyphthalimide
(phthaloxime)
132

I
Indanol
Indanylcarbenicillin
Indomethicin
Inflammation
Injection
direct aqueous I.V. or I.M
of ionic and colloidal iron
pain
vehicle

129
129
76,77
55
66
79
79
60

Intermediates, chemical
116
Intramuscular doses
187
Intraocular pressure
(IOP)
224,227,231,235
Intravenous
administration of hexamethyl
malamine
207
delivery problems of cytotoxic
agents
196
epinephrine to dogs,
233
solutions, problems of
197
Ionization constants, macroscopic
and microscopic
11
Iron, injection of ionic and colloidal 79
L
Latentiation
Leigh's disease
Leprosy
Lincomycin
Linkage, covalent bond
Lipid solubility
Lymph nodes

2
15,21
89
32
3
-9,155
33

M
Mafenide
36,49
acetate
3
Mephenytoin
72, 74, 154
Metabolic pathways
227
Metabolism
during oral absorption
43
enzymatic drug
201
time course of drug
4
Metabolic disposition
184
Metabolites
6, 49, 129
Metal chelating agents
34
Methanamine
1
Methoxymethyl-6-aminopenicillinate.. 122
Methoxymethyl hetacillin 16
122
4"'Methyldigoxin
40
15-Methyl F
44
3-Methylol-5,5-dimethylhydantoin
64
3- ( 4-Methylpiperazinyliminomethyl )
rifamycin SV
137
Milk
21
Modification, bioreversible chemical 73
Modification, covalent chemical
86
Morphine
2,12, 48, 49
2

Naloxone
pamoate
Nandrolone
decanoate
phenylpropionate
Narcotic antagonist

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

47,62
62
54
54
54
62

243

INDEX

Neoplastic diseases
Nirvanol
Nitrofuran
Nitrofurantoin
1- ( 5-Nitro-2-furfurylindene )
aminohydantoin
Nonsteroidal anti-inflammatory agents
Norepinephrine
Nucleosides and nucleotides

23
180
154
178

Pivaloyloxymethyl 6-aminopenicillinate
118
Pivaloyloxymethyl-7- ( D-a-aminophenylacetamido ) cephalosporanate
134
154 Pivaloyloxymethyl-7-(D--azidophen76
ylacetamido) cephalosporanate .. 134
14 Pivaloyloxymethyl benzylpenicillinate 118
23 Pivampicillin
119-121
Plasma

conversion of pro-DPH to
DPH in
187,188
N -Octanoyl derivatives
25
level, indomethicin
77
Ocular side effects of epinephrine .... 229
levels of pro-DPH and DPH in
Oleandomycin
30
rats and dogs
187-191
Opthalmic absorption
35
26
Opthalmic delivery
66 Polar compounds
Polar drugs
33,35
Oral
absorption
26
activity
45 Potassium
D-a-azidobenzylpenicillinate
118
bioavailability
30
7- ( D-a-azidophenylacetamido )
-fluphenthixol dihydrochloride
57
cephalosporanate
134
Organ, target
6
benzylpenicillinate
118
Oxazepame
71,90
7-(2'thienylaeetamido) cephalosodium succinate
71
sporadesate
132
Oxyphenisatin
84
Prednisolone
67
acetonide
45

cyclopentylidenedioxy
45
Pain, injection
79
-21-disodium phosphate
67
O-Palmitoylamodiaquine
59
hemisuccinate sodium salt
67
Pamoate salt of cycloguanil
60
-21-m-sulfobenzoate sodium salt .... 67
Parent
74
catecholamine
14 Primacolone
1
compound
2 Pro-agent
3, 64, 86
molecule
117 Procaine
164,165
Parenteral repository pro-drug
59 Pro-DPH
conversion of
165-169,185,187,188
Parenteral solutions ...
196
Parkinson s disease
9
in dogs, plasma levels of
190,191
Patient acceptance of drugs
80
in dogs and rats, tissue levels of
193
intramuscular doses of
187
Penicillin
3,118
properties of
184
delivery of
3
in rats, plasma levels
G
86,130
of
187,188,189,191
pro-drugs
130
salicylate
170,171
salts of
86
separation of
186
V
130
sulfate
190,192
Percorten
55
turbidity times for
168
Percutaneous absorption
35
Permanganate oxidation technique
185 Pro-drug(s)
Peroral administration
190
accidental
90
Pharmaceutical phase
4
approach to antibiotics
7,116,199
Pharmacodynamic phase
4
definition of
1
Pharmacokinetic phase
4
in the formulation of cytotoxic
Phenobarbitone
74
agents
196
Phenylbutazone
78
to increase the aqueous solubility
Phenylmalonic acid, indanyl ester of 125
of a drug
66
Phenytoin
154,184
to lower the toxicity of a drug
73
Physiological
fluids
60
metabolite
6
Pilocarpine
225
parenteral repository
59
Pipothiazine
57
rationale for use of
4
Pivaloyloxymethyl-D-a-aminobenzylretrospective
2
penicillinate
118,131
for sustained or prolonged release 51
6

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

244

PRO-DRUGS

Prontosil
Propoxyphene
Prostaglandins
Psicofuranine
Psoriasis
Psychotic behavior
Pulmonary tissue
Pupillary dilation
Pyridoxine (vitamin B )
palmitate esters of
6

Q
Quaternary
ammonium drugs
compounds
derivatives
Quaternized nitrogen

90
86
45
24
23
55
33
231
15
22

26
26,29
28
17

R
Rabbits, IOP effects in
231
Rate constant, first order
206
Rate determining step
173
Rats
bioactivities in blood of
147
plasma levels of pro-DPH and
DPH in
187-189, 191
tissue levels of pro-DPH and
DPH in
193
urinary excretion of DPH and
pro-DPH sulfate in
192
Release, sustained or prolonged
51
Repository pro-drug effect
59
Retina
226
Retrospective pro-drugs
2
Reversible alteration of drug
properties
196
Riboflavin (vitamin B2)
15
fatty acid esters of
22
Rifampicin
137
S

Salicylic acid
76
derivatives
1, 76
Salt(s)
of antibiotics, macromolecular
33
drug
3
sparingly water soluble
63
Scale-up of derivative, convenient .... 117
Selenium sulfide
85
Serum clindamycin bioactivity
concentrations
145
Serum levels
24,140,148,149
Shock treatment
15
Side effects
207,229
Site-specific delivery
83
Skin conditions
38
Sodium
ampicillin
122
O-benzoyl-7- ( 2thienylacetamido )
cephalosporadesate
132
/

Sodium (continued)
6- ( -phenoxyacetamido )
penicillinate
122
salt of chloramphenical
monosuccinate
71
Solid state degradations
88
Solubility aqueous
( See Aqueous Solubility and Water
soluble )
of drug in physiologicalfluidsand
in injection vehicle
60
lipid
9,155
Sparingly water soluble drugs
63, 66
Stability
of carbenicillin esters
126
of derivative
117
problems of drugs
67,85
bioavailability of
45
dihydroxy groups in
38
succinate esters of
67
therapy
52
Subacute necrotising encephalomyleopathy (SNE)
15,21
Substituted 7-/?-aminocephemol-4carboxylic acid
139
7-Substituted cephalosporanic acid .... 139
21-Succinate esters
67
6- ( D-a-Sulfoaminophenylacetamido )
penicillin
130
Sulfamylon
36
hydrochloride solution
36
Sulfanilamide
90
Sulfonamide antimicrobial agents .... 90
Systemic side effects of epinephrine 229
Target organ, barriers of drug to
6
Taste problem of drugs
80, 81
Temporary transport forms
2
Testosterone
-cyclopentylpropionate ester of
53
decanoylacetate
54
esters
24,52-54
phenyl propionate
54
propionate
54
2-Tetrahydropyranyl-4-allopurinyl
ether
41
Thiamine (vitamin B i )
15,26
blood levels in alcoholic patients .... 19
CNS absorption of
15
levels in milk and blood
21
pro-drugs
89
propyldisulfide
16
tetrafurfuryldisulfide
16
N- [7- ( 2'-Thienylacetamido ) cephalosporanoyloxy] phthalimide
132
7- ( 2'-Thienylacetamide ) cephalo
sporanic acid monoisobutylcarbonate anhydride
132

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

245

INDEX

7- ( 2'-Thienylacetamido ) -3-cyclobutylcarbonyloxymethyl-cephem-4-carboxylic acid


128
7- ( 2'-niienylacetamido ) -3-cyelobutylcarbonyloxymethyl-eephem-4-carboxylic acid
128
7- ( 2'-Thienylacetamido ) -3-cyclobutylcarbonyloxymethyl-cephem-l-oxide-4-carboxylic acid 128
7- ( 2'-Thienylacetamido ) -3-hydroxymethyl-A-cephem-4-carboxylic
acid
128
6-(3-Thienyloxyacetamido) penicillin 131
Thiolate ion
16
Time course of absorption, distribu
tion, metabolism, and excretion
of drugs
4
Time profile of a drug
4
Tissue distribution
75,190
Tissue levels of pro-DPH and DPH
in dogs and rats
193
Topical corticosteroids
38
Toxicity, drug
73, 75
Transport
forms, temporary
2
medium for iron
79
moiety "C"
2
Triacetin
64
2',3',5'-Triacetyl-6-azauridine
24
Triacetyloleandomycin
30
Triamcinolone
37
acetonide
37, 45
2',3,5'-Tribenzoyl-6-azauridine
24
Trichloroethanol
81
Trichloroethyl-4-acetamidophenyl
carbonate
82
9a,ll^,21-Trichloro-16a,17-(isopropylidenedioxy ) -1,4,6-pregnatriene-3,20-dione
47
2

Triclorphos
Triggering mechanisms
Tuberculosis
Tumors, solid
Turbidity times
Tyramine
Tyrosine ethyl ester

81
199
89
207
168
12
12

Ulceration, gastric
Ureido anion
Urinary excretion of DPH and proDPH sulfate in rats
Urine, indanol metabolites in dog ....

Vasoconstrictive mechanism
Vitamin
A

B
Bo
C
D
water soluble
2

75
173
192
129

225
88
15,89
15
15
88
88
15

W
Water solubility and taste of drugs .... 81
Water soluble
bioreversible pro-drug of DPH
175
drugs, sparingly
66
gentisate complex of hexamethyl
melamine
3
salts, sparingly
63
vitamins
15
Wernicke's disease
17

In Pro-drugs as Novel Drug Delivery Systems; Higuchi, T., et al.;


ACS Symposium Series; American Chemical Society: Washington, DC, 1975.

Você também pode gostar