Escolar Documentos
Profissional Documentos
Cultura Documentos
, 50,
677( 1967).
(1) R. I. Poust and V. F. Smolen, J. Pharm. Sci., 59, 1461 (16) M. M. Breuer, J. Phys. Chem., 68,2067(1964).
(1970). (17) Ibid., 68, 2074(1964).
(2) V. F. Smolen and R. 1. Poust, ibid., 60,1745(1971). (18) Ibid., 68,2081(1964).
(3) V. F. Smolen and F. P. Siegel, ibid., 57,378(1968).
(4) V. F. Smolen and L. D. Grimwood, J . Colloid Interface Sci., ACKNOWLEDGMENTSAND ADDRESSES
36, 308(1971).
( 5 ) V. F. Smolen, D. E. Snyder, and R. J. Erb, J . Pharm. Sci., Received January 31, 1972, from the Biophysical Pliarniaceutics
59, 1093(1970). Area of the Department of Industrial and Physical Pharmacy,
(6) V. F. Smolen, Amer. J . Pharm. Educ., 33,381(1969). School of Pharmacy and Pharmacal Sciences, Purdue Unicersity,
(7) V. F. Smolen and E. J. Williams, J . Pharm. Sci., 61, 921 Lafayeite, IN 47907
(1972). Accepted for publication March 7, 1972.
(8) D. S . Riggs, “The Mathematical Approach to Physiological Presented to the Pharmacology and loxicology Section, APHA
Problems,” Williams & Wilkins, Baltimore, hld., 1963, pp. Academy of Pharmaceutical Sciences, Washington, D. C., meeting,
120-168. April 1970.
(9) V. F. Smolen, J. Pharm. Sci., 60,354(1971). Abstracted in part from a thesis prepared by R. I. Poust in partial
(10) R. D. Schoenwald, Ph.D. thesis, Purdue University, Lafay- fulfillment of the Doctor of Philosophy degree requirements.
ette, Ind., 1971. Supported by a grant from Armour-Dial, Inc., Chicago, Ill.
(11) E. Ackerman and J. B. Hazelrig, U. S. Atomic Energy The technical assistance of Mr. Michael Crosby and Mr. Richard
Commission Symposium No. 3, June 1964. Shutt is gratefully acknowledged.
(12) A. R. Cade, Soap Sanit. Chem., 26, 35(1950). * Present address : Department of Pharmaceutics, School of
(13) A. R. Cade, J. Soc. C o m e t . Chem., 2,281(1951). Pharmacy, University of Pittsburgh, Pittsburgh, PA 15213
(14) P. B. Price, Ann. Surg., 134,476(1951). A To whom inquiries should be directed.
Abstract 0 The lack of detailed information on the manipulation oil (5-10 %) plus polyvinylpyrrolidone (4-5 %) containing approxi-
and preparation of cannabinoid formulations prompted an in- mately 1 % cannabinoid. Such an approach incorporated an in-
vestigation of useful vehicles for administration of tetrahydro- nocuous vehicle, did not require the presence or removal of an
cannabinols and crude marijuana extracts. It was found that pure organic solvent, provided wide latitude for needed concentrations
A9- and As-tetrahydrocannabinols could be quantitatively handled of cannabinoid, and was timesaving. The ratio of emulsifier and
by chipping samples at 4“, transferring them to cold receptacles cannabinoid was critical for stable emulsions.
for weighing, and, after liquefying the cannabinoid at 50”, adding a Keyphrases 0 Marijuana constituents-oral and parenteral for-
warmed vehicle for further transfers and final dilution. Tetrahy- mulations, stability in various solvents, biological evaluation of
drocannabinol samples larger than 10 g. were liquefied at 55” and formulations, vehicle toxicity 0 Toxicity, vehicle-stability of
poured directly into a tared receptacle. Crude marijuana extract marijuana constituents in various solvents, biological evaluation of
samples were smeared on tared receptacles and diluted and trans- formulations for long-term oral and parenteral administration to
ferred as above. Stock solutions of cannabinoid in sesame oil were laboratory animals 0 Formulations-effect of solvents on mari-
stable for months and could be used directly for oral administra- juana constituent stability, biological evaluation, vehicle toxicity Ji
tion or for formulating injectables. Suitable emulsions for paren- Tetrahydrocannabinols-stability in various solvents, biological
teral use consisted of sesame oil (10-15%7,)plus polysorbate 80 (0.4- evaluation of formulations for long-term oral and parenteral ad-
1 %) in saline containing up to 4 % tetrahydrocannabinol or sesame ministration, vehicle toxicity
A full understanding of the biological properties of The tetrahydrocannabinols have been identified as
marijuana constituents has been hampered by the lack the major biologically active components of marijuana
of a desirable vehicle for the preparation of injectables. (1, 2). Whereas crude marijuana extracts are of a tar
The diversity of formulations and routes of administra- consistency, the tetrahydrocannabinols are highly vis-
tion of the active ingredients of marijuana has somewhat cous oils, virtually of a glue nature at room temperature.
complicated the comparison of pharmacological, toxi- Despite reports of extensive analytical data to establish
cological, and behavioral data from different labora- the purity of the sample, little detail has been given as to
tories. Those studies involving a single administration how t o transfer and manipulate the compounds to ob-
have justifiably not been too concerned with the influ- tain accurate concentrations.
ence of the vehicle. However, chronic investigations at Relatively high concentrations of tetrahydrocanna-
relatively high doses (>50 mg./kg.) of cannabinoids binol or crude marijuana extract for intragastric use
cannot overlook the effect of the diluent. can be achieved in natural vegetable oils (3). On the
other hand, aqueous systems necessary for parenteral of sesame oil USP, propylene glycol, and glycerol in conjunction
administration must incorporate some type of emulsi- with 1 g. of cannabinoid.
Cannabinoid Solution Preparation-True solutions of canna-
fier. The limitations of such parenteral formulations binoid were prepared by dissolving 1 g. of liquified tetrahydro-
reside in three considerations: (a) the low concentration cannabinol in 1 ml. of ethanol or acetone. Stock solutions in sesame
of cannabinoid achieved dictates the volume of formula- oil were prepared by dissolving 10 g. of liquified cannabinoid in 25
tion that must be given, (b) the concentration of emul- ml. of warmed sesame oil. For all true solutions, formulations were
sifier and number of treatments introduce the hazard transferred by pipet or poured into graduate cylinders t o accom-
plish quantitative dilution at room temperature. In this manner,
of vehicle toxicity, and (c) the lability of the formulation 140% solutions of cannabinoids in ethanol and sesame oil were
may require frequent preparation of the injectable. prepared and sealed in vials flushed with nitrogen.
One must also keep in mind that formulations developed Cannabinoid Emulsion Preparation-One milliliter of 100-400
for animal trials may not be acceptable for human in- mg./ml. of tetrahydrocannabinol in sesame oil was introduced into
a 50-ml. beaker or conical flask followed by 0.01-0.1 ml. of poly-
vestigations. sorbate 802and isotonic saline to a final volume of 10 ml. Emulsifica-
The present paper provides sufficient details as to the tion was performed by sonication with a sonifier at intensity position 5
preparation of dosage forms of tetrahydrocannabinol at 7.5 amp. for 30 sec. 3. On occasion, the polysorbate 80 was replaced
and crude marijuana extract, outlines the limitations of by 5 % polyvinylpyrrolidone in isotonic saline or 1 % poloxalene4.
various injectables, and reviews formulations used in Other emulsions were formed by addition of 0.1-0.3 ml. of warmed
polysorbate 80 to 100-200 mg. of liquified cannabinoid, mixing
marijuana studies. with a microspatula and mechanical Vortex, and followed by tri-
turation with 1-15 ml. of isotonic saline. Attempted emulsion forma-
EXPERIMENTAL tion with cannabinoid solutions in organic solvents was performed
by trituration of 0.1-0.3 ml. of approximately 50% cannabinoid
Cannabinoids-Crude marijuana extract, synthetic (-))-trans- solutions with approximately 0.9 ml. of steroid-suspending vehicle,
A9-tetrahydrocannabinol, and synthetic ( -)-rruns-As-tetrahydro- 10% dextrans, 10% mannitol, or 0.9% saline.
cannabinol were used'. Cannabinoid shipments were in approxi- Stability Studies-Optical rotation of tetrahydrocannabinols
mately 50-g. quantities per bottle, and the crude extract came in was determined at a concentration of 20 mg./ml. in chloroform with
500-g. quantities per bottle. The provided analytical data on all AS- a polarimeter6. Suitable aliquots of ethanol solutions were diluted
tetrahydrocannabinol lot numbers showed a purity between 95 and with chloroform t o attain the necessary analytical concentration.
97% by G C and an optical rotation range of from -163 to -175". In the instance of sesame oil formulations, 10-30 ml. was extracted
Corresponding data for As-tetrahydrocannabinol were 98-99 with an equal volume of h e x a n e 4 z sodium hydroxide in 50%
purity by GC and optical rotation values from -254 to -268". ethanol. Aliquots of the hexane layer were concentrated and ap-
The crude marijuana extract assayed as 25 % Ag-tetrahydrocan- plied to thin-layer silica gel flexible sheets impregnated with di-
nabinol, 2-3% cannabidiol, and 2-3 % cannabinol. methylformamide and developed with hexane-benzene-methanol
Cannabinoid Transfer Procedure-Samples of tetrahydrocan- (30:30:4) in a sandwich chamber for 45 m i t ~ . The ~ . cannabinoids
nabinols between 0.1 and 10 g. were chipped free a t 4" with a clean, were visualized with Turnbull's blue reagent.
cold, ice pick and transferred to cold, tared, glass receptacles. An Biological Studies-Potential cannabinoid vehicles were tested
18 X 60-mm. conical tube was used for samples under 1 g., a 10-ml. in animal trials for clinical toxicity signs. Single treatments of eth-
conical flask was used for samples between 1 and 3 g., and a 2 5-ml anol were given at doses between 0.005 and 0.05 ml./kg. p.0. or
conical flask was used for samples between 3 and 10 g. Weighings i.v. to 6-10 rats, at 2 ml./kg. p.0. to three rhesus monkeys, at 0.45
were rapidly performed on an analytical balance at room temperature. ml./kg. i.v. t o five rhesus monkeys, at 0.75 ml./kg. p.0. to two
The cannabinoid in its receptacle was next placed in a water bath beagle dogs, and at 0.2 ml./kg. i.v. t o three beagle dogs. Chronic
at 45" under a stream of nitrogen until liquefaction occurred ( 5 1 0 treatment with 100% sesame oil was performed at 0.05 ml./kg.
min.). Ten 50-g. samples of tetrahydrocannabinol were directly p.0. for 119 days in 120 rats, at 1 ml./kg. S.C.for 28 days in three
liquified in their shipping container by heating the inverted con- New Zealand rabbits, and at 3 ml./kg. p.0. in eight rhesus monkeys.
tainer with an IR lamp to 55" under a canopy of nitrogen whenever Sesame oil (10%)-polysorbate 80 (0.4%) dispersed in isotonic
possible. The compound was collected in a tared beaker placed in a saline was administered at 0.3 ml./kg. i.p. for 5 days to three rats,
water bath at 50". The crude marijuana extract was also directly at 0.1 ml./kg. i.v. for 5 days to three rats, and at 6 ml./kg. i.v. for 28
warmed in its shipping container to 45" in a water bath, and 5@500 days to two rhesus monkeys. Sesame oil (IOZ) in 4.5% polyvinyl-
g. of liquified material was transferred by pouring into a tared beaker. pyrrolidone was tested at 6 ml./kg. i.v. for 28 days in two rhesus
Samples less than 50 g. were transferred by spatula.
Cannabinoid Solubility Studies-In the solubility trials, 0.1 -ml.
aliquots of analytical reagent grade ethyl alcohol, benzyl alcohol, * Tween 80.
3 Branson, model LS75, Heat Systems-Ultrasonics, Inc.. Plainview.
acetone, or dimethyl sulfoxide were added stepwise t o 1 g. of can- N. Y.
nabinoid until maximum solubility was achieved a t room tempera- 4 Pluronic F68, BASF Wyandotte Corp., Wyandotte, Mich.
ture. Similar solubility studies were conducted with 0.1-ml. aliquots The dextran was obtained from Bios Laboratories, Inc., New York,
N. Y . ;particle size not given.
Bendix Corp., Cincinnati, Ohio.
Supplied under contract with the National Institute of Mental 7 M. Hagopian, Mason Research Institute, Worcester, Mass., un-
Health. published data.
Final Concentration of
-Ingredient (v/v+
Dil-
-Cannabinoid--. -----Solvent- -----Diluent- THC, Solvent, uent,
N.me mg. Name ml. Name ml. Appearance of Preparation mg./ml.
monkeys. Saline solutions of 2.5-10% polysorbate 80 were ad- circumvented by performing transfers of samples less than 10 g. at
ministered at doses between 0.03 ml./kg. i.v. and 0.3 ml./kg. p.0. to 4" and larger samples after liquefaction. Concentrated solutions
5-10 rats; at 2-3.6 ml./kg. p.0. and i.v., respectively, to two rhesus could be quantitatively transferred if the outside walls of receptacles
monkeys, and at 1 ml./kg. i.v. to two beagle dogs. A 5.5% sodium were also warmed.
glycocholate solution was also tested at 2 ml./kg. i.v. for 9 days in Solubility of Cannabinoids in Solvents Yielding Solutions-
two rhesus monkeys. Hematological and clinical chemical param- Because of fragmentary data on the solubility of cannabinoids,
eters were determined by standard procedures and will be the some studies on the solubility of A'- and As-tetrahydrocannabinols
subject of a separate report. and crude marijuana extract were carried out. Tetrahydrocanna-
binols behave primarily as liquids and are freely miscible with many
RESULTS organic solvents. In such cases the limit of solubility may be taken
at that point where the solvent is still in excess. Since the volume
As the tetrahydrocannabinols thawed, they became tacky and contribution by the cannabinoid mass was approximately 80% in
glued to glass, metal, or paper surfaces. This difficulty could be the presence of an organic solvent like ethanol, the solubility limit
Estimated
Tehahydro- Optical Percent
cannabinol, Elapsed Time, Rotation in Purity by
Cannabinoid Diluent mg./ml. Temperature Days Chloroform TLCa
Ag-Tetrahydro- None 1000 5" 0 -163 -
cannabinol None 1000 55" 0 . 5 hr. -160 -
Ethanol 500 5" 0 -164 90
Ethanol 500 5" 40 -152 80
Ethanol 500 22" 40 -126 -
Ethanol 500 5" 90 - 70
Chloroform 20 22" 21 -163 -
Chloroform 20 55O 0 . 5 hr. -161 -
Sesame oil 100 60 - 88
Sesame oil +
polysorbate
10
5 O
5" 7 - 90
Number
Animals
Reacting/
Dose, Number
ml./kg. x Animals
Vehicle Species" Route Days Treated Clinical Signs
of cannabinoid in ethanol may be considered as 1 g./ml. (e.g., the successful emulsions comprised I-ml. aliquots of stock solutions of
ethanol volume was slightly in excess of that of the cannabinoid). cannabinoid in sesame oil ( l W 4 0 0 mg./inl.) to which 0.01 -0.1 ml.
Further additions of cannabinoid to 1 ml. of ethanol yielded true of polysorbate 80 and 8 9 ml. of isotonic saline were added and
solutions, but now the cannabinoid was the solvent and ethanol the emulsified by sonication. Increased amounts of polysorbate 80
solute. In effect, a solurion of 0.95 g./ml. of cannabinoid in ethanol permitted higher concentrations of cannabinoid. Other usable
could be produced by dissolving 1 ml. of ethanol in 3.2 ml. (4 g.) of emulsions were obtained with cannabinoid in sesame oil and 5 10%
canna binoid. polyvinylpyrrolidone or 1 ?< poloxalene'. The sesame oil-poly-
The data in Table 1 are presented for the comparison of canna- vinylpyrrolidone emulsions were stable for 3-6 hr. when a slight
binoid solubility in various solvents equated to I g./ml. of tetra- (5-10%) oil slick formed. This oil slick could be readily reeniulsitied
hydrocannabinol in ethanol. Similar solubiliiies were obtained for by sonication, and this procedure was successfully applied for several
AB- and ~n-tetrahydrocannabinols and crude marijuana extract in days without deterioration of the emulsion. Concentrations of
polar solvents. For practical purposes, 100-400 mg. of cannabinoid polyvinylpyrrolidone above 307; when mixed with cannabinoid in
can be dissolved in I ml. of a suitable organic solvent or natural oil sesame oil yielded two-phase systems. and polyvinylpyrrolidone
like sesame oil. concentrations below 5 % formed unstable emulsions. In the in-
Characteristics of Emulsions-To explore possible aqueous sys- stance of poloxalene, concentrations above 2 7; resulted in unstable
tems for cannabinoids, various combinations of solvents, emulsifiers, emulsions.
and diluents were examined. These results (Table 11) show that Stability of Cannabinoid Formulations- -Because of the time
small quantitites of solubilizer or emulsifier provided stable emul- required to manipulate thc cannabinoid (e.g., chipping, weighing,
sions. Relatively high concentrations of cannabinoid with 10 liquefaction, admixture, and emulsification) and the desire to use
polysorbate 80 yielded emulsions which were stable for a few min- stock solutions for long periods of time, some exploration of chem-
utes; as the concentration of cannabinoid was reduced, the physical ical stability was performcd. These data, presented in Table 111,
stability of the emulsion increased. Reduction of the polysorbate 80 reveal that the optical rotations of the tetrahydrocannabinols were
content severely restricted the amount of cannabinoid that could not affected by elevated temperatures for short periods of time. In
be stably emulsified. ethanolic solutions at concentrations from 500 to loo0 mg./ml., the
Small quantities of organic solvents failed to provide suitable optical density of Sg-tetrahydrocannabinol decreased about 8%
cannabinoid suspensions (c.g., propylene glycol and glycerol after 40 days at 4". The decrement was more noticeable over this
formed a two-phase system upon dilution with saline). The most interval of time at room temperature. Under similar conditions, the
8 Communication obtained under National Institute of Mental Health 1-1 Triton X-100.
contract elsewhere. “These vehicles were tested in collaboration with Dr. Monroe E.
9 Unpublished data. Wall.
Abstract 0 A series of solid, substituted benzoic acids (p-XC&- Guillory and Higuchi (29).
COOH), which decompose into a liquid (xC6I-h) and carbon diox-
ide, were studied. For decomposition to take place below the melt- Attempts to correlate decompositions in the solid
ing point, the u value must be less than -0.35; the decomposition state with usual substituent parameters in homologous
then follows Bawn-type kinetics. Neither solid (ks) nor liquid (kz) series have not met with success. Dorko et a2. (30)
decomposition constants show isokinetic relations at their melting found no such correlation in a study of substituted
points. However, log k , is proportional to l/Tm,where T , is the
tosylates, and Meyers et al. (31) found that in the re-
absolute melting temperature, much as was found for vitamin A
esters. action R’COONa +R”C0OH + R’COOH +
Key phrases Decomposition rates of solid compounds-p-sub- R”COONa, substituent u values were the governing
stituted benzoic acid derivatives 0 Solid-state decomposition- parameters in that u’ would have to be larger than u”
p-substituted benzoic acid derivatives 0Benzoic acid derivatives- for the reaction to occur; this, in essence, is paramount
solid-state decomposition, rate constants to the time-tested rule that “the stronger acid drives
~
out the weaker acid.” The latter two studies aimed at the
Many studies have been conducted related to decom- importance of the chemical factors involved in reactivity
position rates of solid compounds. A great majority in the solid state; whereas, in general, physical pa-
of these have concerned inorganic salt decompositions rameters (active sites, dislocations, efc.) have been the
[carbonates (1-9), oxalates (1&17), and permanganates bases for proposed hypotheses.
(18-21)], and several general patterns have been pro- The pharmaceutical literature has partially touched
posed as primary mechanisms in the decompositions; upon the importance of liquid layers as mediators of
most notable of these are the Prout-Tompkins model the actual decompositions (26, 29), whereas the re-
(21) and the power laws (22-25). Some studies have maining great majority of the cases cited dealt with re-
been reported in the pharmaceutical literature, notably actions of the type solid + solid +
gas. The work deal-
the ones by Leeson and Mattocks (26), by Kornblum ing with aspirin anhydride (28), as well as the work