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Article history: The purpose of the present work was to provide theoretical and experimental support in generating an
Received 10 June 2009 optimal pH (pHmax ) for a representative weak base compound (propranolol), that can lead to enhanced
Received in revised form 23 October 2009 sublingual absorption. Initially equations for pH-solubility and pH-permeability profiles were derived
Accepted 23 December 2009
and compared to the profiles obtained experimentally. Excellent correlation (R2 = 0.999) of solubility
Available online 8 January 2010
profiles was obtained using non-linear regression, and the permeability profiles further predicted that at
certain pH (pHmax ), optimal mucosal permeation could be achieved. Subsequently, in a pharmacokinetics
Keywords:
study, a buffered sublingual propranolol tablet, designed to achieve its pHmax (when dissolved in saliva),
Sublingual
Weak base
were compared to that from a marketed product (Inderal® which could not achieve pHmax ) in 8 healthy
Propranolol subjects. Each subject received the products sublingually for 15 min followed by swallowing the remaining
Permeability drug–saliva. The plasma propranolol concentrations of AUC during first 30 min from the buffered tablet
Pharmacokinetics were significantly higher than that from the Inderal® tablet (p < 0.05), and no significant differences in the
remaining AUC were observed. These in vitro and in vivo results on propranolol provided experimental
confirmation of the pHmax concept as well as its utility in sublingual drug delivery. Such an approach may
be applicable to other similar compounds to improve sublingual drug delivery.
© 2010 Elsevier B.V. All rights reserved.
0928-0987/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.ejps.2009.12.011
Y. Wang et al. / European Journal of Pharmaceutical Sciences 39 (2010) 272–278 273
ity than at any other pH. The aims of this paper are (1) to derive steady-state flux (amount permeated across the membrane per
the quantitative relationship of pHmax , solubility and permeabil- unit time and area) of the ionized (Jss(i) ), unionized (Jss(u) ), and total
ity, and (2) to demonstrate the application of its pHmax concept in species (Jss(T) ) at a given constant pH solution can be expressed
improving sublingual absorption or permeation of a representative using Fick’s second law:
weak base compound, propranolol, using laboratory and human
dQi
experiments. Jss(i) = = Pi Ci (7)
dt · A
2. Theory dQu
Jss(u) = = Pu Cu (8)
dt · A
The theoretical equations describing pH-solubility and pH- dQ
permeability (flux) profiles are shown bellow. Jss(T ) = = Jss(u) + Jss(i) = Pi Ci + Pu Cu (9)
dt · A
where Qi , Qu and Q are the accumulated amount of the ion-
2.1. pH and solubility profile of weak base
ized, unionized and total species permeated across the membrane,
respectively. A is the permeation area, Ci and Cu are the soluble
When a weak base is dissolved in an aqueous solution, equilib-
concentrations of ionized and unionized species, respectively, and
rium exists between the unionized species (B) and ionized species
Pi and Pu are the individual permeability coefficients of ionized and
(BH+ ), and can be described by Eq. (1):
unionized species, respectively. (Pi and Pu should be independent
Ka of solution pH provided that the structure and physicochemical
BH+ + H2 OB + H3 O+ (1)
properties of each species are not influenced by pH).
where Ka is the dissociation constant of the weak base and can be Theoretically, at pH = pHmax , both Ci and Cu will be saturated or
expressed as: maximized (i.e. equal to Si and Su , respectively), therefore, the high-
est Jss(T) can be expected. Jss(T) values at pH > pHmax and pH < pHmax
[B][H3 O+ ] Cu [H3 O+ ]
Ka = + or (2) can be obtained using Pi and Pu and Ci and Cu (or Si , Su ) from Eqs.
BH Ci
(10) and (11) below.At pH ≤ pHmax
where [B] or Cu is the concentration of the unionized species and
Ka
[BH+ ] or Ci is the concentration of the ionized species. Eq. (2) can Jss(T ) = Pi Si + Pu Si (10)
be also re-written as: [H3 O+ ]
2.2. Relation of permeability to solubility at different pHs To determine the solubility of propranolol at different pH values,
an excess amount of propranolol hydrochloride was first added to
For a highly lipid soluble weak base compound like propranolol, tubes containing isotonic phosphate buffer and their pH adjusted
permeation of the total weak base across the lipid membrane is using 85% (w/w) phosphoric acid or 20% (w/v) sodium hydroxide
expected to include both the unionized and ionized species. The (to achieve a pH range of 5.4–10.0). The tubes were then immersed
274 Y. Wang et al. / European Journal of Pharmaceutical Sciences 39 (2010) 272–278
in a shaking water bath at 37 ◦ C for 24 h and the pH of the solu- all subjects underwent ECG and mouth examination and received
tions were verified. The final mixture was filtered through a 0.2 m instruction not to take any medications 2 weeks prior to and during
syringe filter (Iwaki Glass, Japan), and the total concentration of the period of the study. The subject inclusion criteria were: (a) male,
propranolol or ST (in the filtrate) was determined by HPLC using 18–55 years old, (b) body weight within 15% of ideal weight, (c)
the method described by Nielsen and Rassing (2000) with slight accessible vein for blood sampling, (d) high probability for compli-
modification. ance and completion of the study, (e) no significant abnormalities
in general physical examination, (f) ECG recording within normal
3.3. Determination of propranolol permeability limits, and (g) biochemical and haematological parameters within
normal limits.
The permeability experiment was performed using fresh The subject exclusion criteria consisted of: (a) history of hepatic,
porcine sublingual mucosa obtained from local slaughter house renal, biliary, cardiovascular, gastrointestinal, haematological and
(excised from the floor of the mouth of male adult pigs with the other chronic and acute diseases within 3 months prior to the study,
epithelium layer separated from the underlying connective tissue) (b) clinically relevant abnormality in physical examination, ECG
within 2 h of procurement. A side-by-side diffusion chamber sys- evaluation, urine test, blood chemistry or haematological test, (c)
tem (PermeGear Co., PA, USA) was used for the permeability study. tobacco use in any form, (d) regular consumer of alcohol, (e) blood
The temperature of the system was maintained at 37 ◦ C and the donation within 4 weeks prior to the start of the study, (f) treatment
solution (isotonic phosphate buffer) in each chamber was stirred with propranolol or similar class of drugs within 4 weeks before the
with magnetic bars. The sublingual mucosa was then mounted study, (g) participation in any clinical drug study within 2 months
between the donor and receiver chamber with the surface of prior to the study, and (h) hypersensitivity to propranolol or other
mucosa facing the donor chamber. After a 30 min equilibration, the -blockers.
receiver chamber and donor chamber were freshly filled with 4 ml The subjects were randomly assigned to two equal size groups
pre-warmed phosphate buffer and saturated propranolol solution with one group receiving a single 40 mg dose of buffered tablet and
at different specified pHs, respectively. A 200 l sample was subse- the other group receiving a 40 mg Inderal® tablet. After a washout
quently withdrawn from the receiver chamber every 15 min until period of at least 1 week, each subject was crossed over to receive
2 h and the propranolol concentration was determined (by HPLC, the other formulation.
described below). Jss(T) was then obtained according to Eq. (9). Alcohol, caffeine-containing products and smoking were not
permitted for 24 h prior to, and for the duration of, the study. At
3.4. Preparation of special buffered sublingual propranolol tablet the time of the clinical study, each subject was required to place
to enhance permeability the tablet under the tongue and not to swallow any tablet or saliva
until 15 min, at which time the saliva (containing the dissolved
To verify the applicability of the pHmax concept for sublingual propranolol) was swallowed. Venous blood samples (5 ml) were
delivery, a buffered sublingual propranolol tablet was prepared by collected at 0, 3, 6, 10, 15, 30, 45, 60, 90, 150, 300 and 420 min
direct compression of 40 mg propranolol with excipients (disodium after the start of sublingual administration. The samples were cen-
hydrogen phosphate, lactose, hydroxypropyl methylcellulose and trifuged at 2000 rpm for 10 min at 4 ◦ C and then plasma obtained
magnesium stearate). This formulation was designed to achieve a and stored at −80 ◦ C until analysis.
saliva pH of 7.4–7.6, i.e. close to pHmax when dissolved in saliva
under the tongue. In addition, a marketed propranolol formulation 3.6. Propranolol assay
(Inderal® ), which achieved pH 6.7–6.9 when mixed with human
saliva, was selected for comparison. An HPLC system consisted of a Waters 600 controller, a 717plus
Our buffered tablets were prepared according to the fol- auto-sampler, a Waters 2487 dual absorbance detector, Waters
lowing steps: (1) weigh and mix the propranolol powder with 464 fluorescence detector and a Thermo Hypersil-Keystone Col-
magnesium stearate in a V-blender (ERWEKA Co., Germany) for umn (250 mm × 4.6 mm, 5 Hypersil BDS C18 ) was used for the
30 min; (2) add other excipients to the mixed powder prepared assay.
in step (1), and then commingle the powder in the V-blender for Propranolol samples in both solubility and permeability studies
30 min; (3) directly compress the blended composition into plain were determined using Waters 2487 dual absorbance detector,
tablets (100 mg/tab) using a rotary tablet press machine (Mei-King the mobile phase consisted of 35% acetonitrile and 65% 25 mM PBS
Machinery Engineering Co., Hong Kong), with 6 mm in diameter (pH 4.0, containing 0.5% triethylamine) and the samples were run
and 2–3 mm in thickness. Each tablet should contain 40 mg pro- at a flow rate of 1 ml/min. The chromatograms were recorded at
pranolol hydrochloride and 40 mg disodium hydrogen phosphate. 230 nm. Excellent linearity of the calibration curve (R2 = 0.9998)
The tablet hardness, friability, weight variation, content uniformity, within the concentration range studied was obtained. The assay
disintegration time, and dissolution profile were also measured detection limit was 55 ng/ml and the inter-day and intra-day coef-
according to the USP monograph. ficients of variation were less than 5%.
For plasma propranolol concentration determination, 25 l
3.5. Human pharmacokinetic study internal standard (verapamil 40 g/ml), 50 l 2 M NaOH and 5 ml
n-hexane plus isopropyl alcohol mixture were added to 1 ml
To verify the enhanced absorption (permeation) using the plasma sample. After shaking (10 min) and centrifugation (10 min
pHmax concept, a pharmacokinetic study was carried out in 8 at 4000 rpm), the organic phase (4 ml) was transported to a clean
healthy male volunteers (aged 18–30 years with body weight tube and 200 l of 0.05 M H3 PO4 was added. After mixing (1 min)
between 50 and 90 kg) who received the special buffered propra- and centrifugation (2 min at 10800 rpm), the upper organic phase
nolol tablet and Inderal® tablet in a random crossover manner. was removed and 100 l of the aqueous layer was injected into an
The study followed the tenets of the Declaration of Helsinki (1964) HPLC system for analysis.
and the study protocol was approved by the Joint Chinese Univer- The plasma propranolol concentration was determined using
sity of Hong Kong—New Territories East Cluster Clinical Research Waters 464 fluorescence detector. The mobile phase, composed of
and Ethics Committee. The approved Informed Consent Form from 0.05 M KH2 PO4 buffer (pH 3.0, adjusted by 2 M HCl) and methanol
the Committee was signed by all subjects. Prior to the studies, (40:60), was run at a flow rate of 1 ml/min. The assay calibra-
Y. Wang et al. / European Journal of Pharmaceutical Sciences 39 (2010) 272–278 275
Table 1
Unionized fraction of propranolol and its solubility at various pH levels (n = 4).
4. Results Fig. 2. Effect of pH on the steady-state flux of saturated propranolol across porcine
sublingual mucosa.
4.1. pH-solubility profile and pHmax
4.2. pH-permeability profile
The solubilities of propranolol at pH 5.4–10.0 are shown in
Table 1. The pHmax was determined to be 7.62 (see below). The amount of propranolol permeated across the membrane per
The fraction of ionized and unionized propranolol was cal- unit time and area, i.e. Jss(T) , at different pH (determined experimen-
culated according to Eq. (3). At pH 5.4, the total solubility of tally according to using Eq. (9)) were compared to that generated
propranolol was found to be 22.12 mg/ml, with 99.99% of propra- theoretically (Eqs. (10) and (11)) are shown in Figs. 2 and 3,
nolol in the ionized form (Table 1). Therefore, the solubility of the respectively. The maximum Jss values corresponded to pH 7.40
ionized form (Si ) was obtained from the overall solubility of pro-
pranolol at pH 5.4:
Table 2
Pharmacokinetic parameters following sublingual administration of the special
buffered propranolol buffered and Inderal® tablet after in eight male healthy sub-
jects (mean ± SD, n = 8).
that at pHmax . In addition to this in-vivo observation, we found that investigated preliminarily the permeation profile of 2 additional
in a limited number of human subjects, the heart rate reduction fol- weak base compounds, nebivolol and verapamil and confirmed a
lowing sublingual administration of propranolol was greater at a similar close relationship of the theoretical vs. experimental data,
saliva pH maintained around pHmax than that at higher or lower using the pHmax concept illustrated by propranolol.
pH values (unpublished data). Our significantly greater plasma The present pHmax concept, however, is unlikely be applica-
concentration data during initial 15 min following our buffered ble to all weak base compounds. Those weak base compounds
tablet (achieving salvia pH 7.4–7.6) compared to Inderal® (achiev- with low molecular weight, high partition coefficient, and single
ing saliva pH 6.7–6.9) provide further support of the pHmax concept. pKa value may be suitable. For most hydrophilic compounds, or
In the present study, the initial higher plasma concentrations macromolecules (i.e. peptides and proteins), the transmucosal per-
(up to 30 min) following the sublingual administration of our meability of the ionized species are extreme low. Thus, the pHmax
buffered formulation compared to Inderal® tablet is most likely concept cannot be applied for these compounds.
a reflection of the differences in the absorption of the two for-
mulations during the first 15 min of sublingual administration.
6. Conclusion
There were no significant differences in the overall Tmax , Cmax ,
AUC30–420 min and AUC0–420 min values between the two formu-
In summary, the present work demonstrated that the trans-
lations, since these data largely reflected gut absorption after
membrane flux for a weak base compound such as propranolol,
swallowing of the saliva–drug solution at 15 min post-sublingual
across a mucosal membrane, could be enhanced by applying the
administration. The two-peak observation was also found with vin-
pHmax concept, leading to the development of a potential sublin-
camine when administered in a similar manner by others (Aiache
gual product with rapid enhanced absorption. Such a concept may
et al., 1990).
improve the development of certain other weak base compounds
The faster dissolution from our buffered tablet may have
suitable for sublingual or other mucosal administration to achieve
resulted in faster sublingual absorption in first 5–10 min. How-
rapid onset of action for urgent medical conditions that require
ever, the mean propranolol concentration in plasma following
such intervention.
buffered tablets was abut 10-fold higher than that of Inderal®
tablets at 3 min, and about 5.9-fold at 6 min. These data were
disproportional to the 70% higher release from the buffered tablets Acknowledgements
in comparison to the Inderal® tablets at 5 min. Moreover, the
propranolol plasma concentration following buffered tablets was This work was supported by a Direct Grant (No. CUHK 2041010)
1–2 folds higher than Inderal® tablet at 10–30 min, when the drug from The Chinese University of Hong Kong and an ITF Grant (No.
release profiles were similar between the two formulations. Since ITS/174/00) from the Innovation & Technology Commission. The
both buffered and Inderal® tablets were completely dissolved authors are also thankful to Dr. Benny Fok, Mr. W.K. Fung and Ms.
sublingually at about 15 min, it appeared that the major factor Miranda W.S. Lai for their technical and logistic assistance.
contributing to the significantly faster sublingual absorption and
higher propranolol plasma concentration was the optimal saliva References
pH generated by the buffered tablet.
The buffered tablet was designed for rapid disintegration Aiache, J.M., Delatte, M.C., Leblanc, P.P., Kantelip, J.P., Gomeni, R., Steimer, J.L.,
(<1 min). However, due to small volume of saliva generated per 1990. Pharmacokinetic aspects of the sublingual administration of vicamine.
Biopharm. Drug. Dispos. 11, 279–309.
minute, the amount/min that can be disintegrated is small. Nev- Beckett, A.H., Triggs, E.J., 1967. Buccal absorption of basic drugs and its application
ertheless, whatever the amount disintegrated, the buffering agent as an in vivo model of passive drug transfer through lipid membranes. J. Pharm.
will keep the saliva pH near pHmax thus allowing optimal absorp- Pharmacol. 19 (Suppl), 31S–41S.
Beyssac, E., Touaref, F., Meyer, M., Jacob, L., Sandouk, P., Aiache, J.M., 1998. Bioavail-
tion, leading to significantly earlier and higher propranolol concen-
ability of morphine after administration of a new bioadhesive buccal tablet.
trations in first 30 min as compared to the conventional formula- Biopharm. Drug Dispos. 19, 401–405.
tion without the appropriate buffering agent. To further enhance Cardot, J.M., Chaumont, C., Dubray, C., Costantini, D., Aiache, J.M., 2004. Comparison
of the pharmacokinetics of miconazole after administration via a bioadhesive
and prolong mucosal absorption, bioadhesive technology may be
slow release tablet and an oral gel to healthy male and female subjects. Br. J.
an attractive choice by utilizing various bioadhesive polymers, i.e. Clin. Pharmacol. 58, 345–351.
Cabomer, HPMC, and chitosan, etc. in the future (Beyssac et al., Chen, L.L., Chetty, D.J., Chien, Y.W., 1999. A mechanistic analysis to characterize
1998; Yong et al., 2001; Hassan et al., 2009; Cardot et al., 2004). oralmucosal permeation properties. Int. J. Pharm. 184, 63–72.
Chetty, D.J., Chen, L.L., Chien, Y.W., 2001. Characterization of captopril sublingual
The optimization of rapid sublingual absorption of propra- permeation: determination of preferred routes and mechanism. J. Pharm. Sci.
nolol by utilizing the pHmax concept may offer a new therapeutic 90, 1868–1877.
approach to meet its clinical needs. Propranolol is widely used for Galey, W.R., Lonsdale, H.K., Nacht, S., 1976. The in vitro permeability of skin and
buccal mucosa to selected drugs and tritriated water. J. Invest. Dermatol. 67,
the long-term maintenance treatment and prophylaxis of many 713–717.
cardiovascular conditions (Koch-Wesser and Frishman, 1981; Hao, J., Heng, P.W., 2003. Buccal delivery systems. Drug Dev. Ind. Pharm. 29,
Regardh, 1982; Wood et al., 1978). For some of these conditions 821–832.
Hassan, N., Khar, R.K., Ali, M., Ali, J., 2009. Development and evaluation of buc-
which are acute but episodic, e.g. anxiety tachycardia, situational cal bioadhesive tablet of an anti-emetic agent ondansetron. AAPS Pharm. Sci.
tachycardia, paroxysmal atrial fibrillation and tachycardia that Technol. [Epub ahead of print].
can occur suddenly in the ambulatory setting, instead of current Johnsson, G., Regardh, C.G., 1976. Clinical pharmacokinetics of beta-adrenoreceptor
blocking drugs. Clin. Pharmacokinet. 1, 233–263.
approach of oral chronic maintenance or prophylaxis, administer-
Koch-Wesser, J., Frishman, W.H., 1981. -adrenoceptor antagonists new drugs and
ing buffered tablet at the onset of each episode or using “pill in new indications. N. Engl. J. Med. 305, 500–506.
the pocket” approach, may conveniently provide rapid “targeted” Kramer, S.F., Flynn, G.L., 1972. Solubility of organic hydrochlorides. Pharm. Res. 61,
1896–1904.
therapy for these episodes. Also, this can avoid unnecessary drug
Ledwidge, M.T., Corrigan, O.I., 1998. Effects of surface characteristics and solid state
consumption at the time between the episodes, especially if these forms on the pH solubility profiles of drug–slats systems. Int. J. Pharm. 174,
episodes are infrequent. 187–200.
The present study has provided clearly a detailed example of Nielsen, H.M., Rassing, M.R., 2000. TR146 cells grown on filters as a model of human
buccal epithelium: IV. Permeability of water, mannitol, testosterone and beta-
the application of the pHmax concept to permeation of propranolol, adrenoceptor antagonists. Comparison to human, monkey and porcine buccal
a well-known representative weak base compound. We have now mucosa. Int. J. Pharm. 194, 155–167.
278 Y. Wang et al. / European Journal of Pharmaceutical Sciences 39 (2010) 272–278
Nies, A.S., Shand, D.G., 1975. Clinical pharmacology of propranolol. Circulation 52, Taylan, B., Capan, Y., Gueven, O., Kes, S., Atilla, H.A., 1996. Design and evaluation
6–15. of sustained-release and buccal adhesive propranolol hydrochloride tablets. J.
Place, V., Darley, P., Baricevic, K., Ramans, A., Pruitt, B., Guittard, G., 1998. Human Control. Release 38, 11–20.
buccal assay for evaluation of the mucosal irritation potential of drugs. Clin. van de Donk, H.J., Merkus, F.W., 1982. Decreases in ciliary beat frequency due to
Pharmacol. Ther. 43, 233–241. intranasal administration of propranolol. J. Pharm. Sci. 71, 595–596.
Regardh, C.G., 1982. Pharmacokinetic aspects of some beta-adrenoceptor blocking Wood, A.J., Carr, K., Vestal, R.E., Belcher, S., Wilkinson, G.R., Shand, D.G., 1978. Direct
drugs. Acta Med. Scand. 665 (Suppl.), 49–60. measurement of propranolol bioavailability during accumulation to steady-
Schoenwald, R.D., Huang, H.S., 1983. Corneal penetration behavior of -blocking state. Br. J. Clin. Pharmacol. 6, 345–350.
agents. I: Physicochemical factors. J. Pharm. Sci. 72, 1266–1272. Yong, C.S., Jung, J.H., Rhee, J.D., Kim, C.K., Choi, H.G., 2001. Physicochemical char-
Shojaei, A.H., Berner, B., Li, X., 1998. Transbuccal delivery of acyclovir: I. In vitro acterization and evaluation of buccal adhesive tablets containing omeprazole.
determination of routes of buccal transport. Pharm. Res. 15, 1182–1188. Drug Dev. Ind. Pharm. 27, 447–455.
Squier, C.A., Hall, B.K., 1985. In vitro permeability of porcine oral mucosa after
epithelial separation, stripping and hydration. Arch. Oral Biol. 30, 485–491.