Wissenschaft

und Technik:

Driginale

In vitro and in vivo Evaluation of Floating Controlled Release Dosage Forms of Verapamil Hydrochloride
Seham A. Elkheshen*, Alaa Eldeen B. Yassin, SaIeh Alsuwayeh, and Fayza A. AlkhaIed Department of Pharmaceutics, Collage of Pharmacy, King Saud University, Riyadh (Saudi Arabia) Present address: see address for correspondence

Summary
The present work investigates the preparation of sustained release floating systems for verapamll hydrochloride using different hydrocolloid polymers including hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), ethylcellulose (EC) and Carbopol (CP). Floating was achieved by adding an effervescent mixture of sodium bicarbonate and anhydrous citric acid. Some factors were investigated concerning their effect on flotation and rate of drug release including the drug to polymer ratio, the granulating agents, the incorporation of release retardant, coating granules with ethyl cellulose, and finally granule-compression into tablets. A formula composed of 1:1 of verapamil to HPMC, 5 % of gas generating mixture and prepared by wet granulation with 96% alcohol followed by compression was chosen for in-vivo evaluation in comparison with a commercial controlled release product of verapamll hydrochloride. Investigations have been undertaken in beagle dogs to evaluate both the intra-gastric retention performances using X-ray and the bioavallabllity of the drug from both test and standard tablets. Results showed that for powder-blend filled into capsules, only HPMC-4000 formulations combined a good floating and reasonable delay in drug release. However granulation of the same formulations led to complete loss of both the floating and the sustained release characteristics. Tableting of granules containing various verapamil:HPMC-4000 ratios showed excellent buoyancy and slow release prome. Results also revealed that floatation was able to delay the gastric emptying of verapamil tablet in beagle dogs for more than four hours compared to almost one hour for a control tablet devoid of the gel forming polymer and the gas generating mixture. The floating tablets showed bioequivalence with a commercial sustained release tablet with higher mean AUCo~ and Cm~ and longer 1m~, however, non-significantly different.

Zusammenfassung
Untersuchung von flotierenden Formullerungen zur kontrolllerten Freisetzung von Verapamll-Hydrochlorid in vitro und in vivo Untersuchungen ilber die Zubereitung flotierender Systeme ffir die verzOgerte Freisetzung von Verapamil-Hydrochlorid werden beschrieben; verschledene Hydrokolloid-Polymere einschlieBllch Hydroxypropylmehtylcellulose (HPMC), Hyroxypropylcellulose, Ethylcellulose und Carhopol wurden eingesetzt. Flotation wurde durch den Zusatz einer gasbildenden MIschung von Natrlumbicarhonat und wasserfreier Zltronensaure erreicht. Einige Faktoren mit EinfluB auf Flotation und Wirkstofffreisetzung wurden untersucht, u. a. Wirkstoff-Polymer-VerhaIU1is, Granuliermittel, Zusatz von frelsetzungsverwgernden Mlttein, Oberzlehen der Granula mit Ethylcellulose, Verpressen des Granulates zu Tabletten. Filr die In-vivo-Priifung Im Vergleich mit einem kommerziell erhaItlichen Verapamll-Hydrochlorid-Produkt mit kontrollierter Freisetzung wurde eine Formulierung ausgewahlt, die Verapamil und HPMC Im VerhaItnls 1:1 sowie 5 % gasbildende Agentlen enthaIt, mit 96 % Alkohol na~ granullert und anschlie~nd verpreBt wurde. Von beiden Priiparaten
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wurde bei Beagle-Hunden die Bloverftigbarkeit sowie die Relentlon irn Magen ronlgenologisch unlersuchl. Die Ergebnisse zeigen, da~ bei den in Kapseln verftilllen Pulverrnischungen nur soIche rnit HPMC-4000 sowohl Flolatlon als auch annehmbare Verwgerung der FreiselZung ergeben. Die Granulierung derselben Pulverformulierungen fUhrle zu einem vollslandigen Verlusl so-

wohl der Rolalion als auch der Ver:rogerung der FreiselZung. Die Tablellierung von Granulalen rnit verschiedenen Veraparnil-HPl\IC-4000-Verhiiltnissen ergab ausgezeichneles Aufschwimrnen und langsarne FreiselZungsprofile. Durch die Rotalion wurde die Entleerung der Veraparnil-Tablellen aus dern Magen von Beagle-Hunden urn mehr als vier Slunden verwgerl - irn Vergleich zu nahezu einer

Stunde bei einem KonlroUpriiparal ohne gelbildendem Polymer und ohne gasbildende Zusatze. Die Ootlerende Tablellenzubereitung zeigte Bioaquivalenz mit einer handelsiiblichen Tablelle mit ver:rogerler FreiselZung, wobei rniltlere AUCO~, Cmax und ~ nichl signiflkanl hahere Werle hallen.

1. Introduction
Verapamil hydrochloride (VP), the first calcium channel blocker to be approved by FDA in 1981, is useful for the treatment of angina pectoris, hypertension, and supraventricular tachyarrhythmia [I]. In atrial fibrillation, VP is more effective than digoxin for controlling ventricular rate [2]. As being a drug for chronic conditions the extended-release oral dosage form would increase the patient compliance and improve the therapeutic responses by reducing the peak-to-trough variation of drug plasma concentration. The already available extended-release VP per-oral dosage forms, given once daily, are as effective in lowering blood pressure over 24 h as doses of conventional VP formulations given three times daily [3]. VP is considered a good candidate for incorporation in a gastro-retentive dosage form due to its high solubility in the stomach medium compared with its solubility in the small intestine medium [4]. This will allow the gradual release ofVP in solution form to the upper part of the small intestine, where it is mainly absorbed, which will maximize the absorption compared with other controlled release approaches. Floating dosage forms are dosage forms with a bulk density lower than that of the gastric content. This allows them to remain buoyant on the surface of the gastric content for a certain period of time without affecting the intrinsic rate of emptying. They are also referred to as hydrodynamically balanced systems (HBS) as they are able to maintain their low density while the polymer hydrates and builds a gelled barrier at the outer surface [5]. The drug is progressively released from the swollen matrix as in the case of conventional hydrophilic matrices. In order to design successful floating dosage forms, three major conditions should be met: (i) They must have sufficient structure to form a cohesive gel barrier; (ii) they must have an overall specific gravity lower than that of gastric contents (reported as 1.004-1.010 g/m\); (iii) and finally they should dissolve slowly enough to serve as a reservoir for the delivery system [61. The selection of potential excipients that allow the formulation of matrices having sustained delivery characteristics and a bulk density of less than 1 gl ml is the key point. After extensive work on the floating and swelling characteristics of commonly used excipients, Gerogiannis et al. [7J concluded that polymers with
Pharm. Ind. 66, Nr. 11, 1364-1372 (2004)
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high molecular weight and less hydrophilic grades usually exhibit enhanced floating characteristics. Floatation not only prolongs the gastrointestinal (GI) residence time, or obtains a sustained local action into the stomach, but also does so in an area of the gastrointestinal tract (GIT) that would maximize the amount of the drug reaching its absorption site in solution, and hence ready for absorption. The retentive characteristics of the dosage form are most significant for drugs insoluble in intestinal fluids, for those acting locally in the upper GIT and those exhibiting site-specific absorption [6]. The techniques applied to achieve floatation are numerous. The design of an inflatable chamber containing a liquid that gasifies at body temperature is an example [8]. The preparation of an empty globular shell covered by a solid formulation is another example [9]. Harrigan [10] designed a floating dosage form by the incorporation of a floatation chamber filled with air or a harmless gas. The incorporation of fatty materials, having bulk density less than 1 g/ml, with hydrocolloid to decrease water uptake and increase the duration of buoyancy was investigated [U]. The chemical generation of CO2 by incorporation of an acid and a carbonate salt that will react in the presence of the aqueous stomach medium is a frequently used method for the induction of floatation [12, 13]. Different hydrocolloids of natural and semisynthetic origin have been used for the formulations of HBS forms [14, IS]. Among the different hydrocolloids recommended for floating formulation, cellulose ether polymers are the first choice, especially hydroxypropylmethylcellulose (HPMC), which is extensively used by many authors [16-18]. The present work involves tile use of several polymers such as HPMC-4000 (hydroxypropylmethylcellulose), HPC-MF (hydroxypropylcellulose), EC-25 (ethylcellulose), CP-934p (Carbo po\) and a number of formulation metllOds as gran ulation, film coating of the granu~es and compression of granules into tablets in order to develop a floating solid dosage form for controlling the release of verapamil hydrochloride (VP). A formula that combined both excellent floating and sustained release characteristics was chosen for farther in vivo evaluation in comparison with a commercial controlled release product of verapamil hydrochloride.

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2. Experimental F7 F4 F2 F3 Fs Fs
2.1. Materials F,

size 18 48 9 120 120 7 2 12 24 96 120 6 0 0 1 12 120 240 18 9 00 1 Table1212 1: Composition Capsule der mix (mg). CA NaHCOj

----

of verapamIl capsules filled with powGG

Verapamil HCl from Adwic Chemical Co. (Cairo, Egypt), Hydroxypropylmethylcellulose 4000 from Shin-Etsu chemicals (Tokyo, Japan). hydroxypropylcellulose, Kiucel MF from AquaIon Co. (Delaware, USA), ethylcellulose USP/NF from Spectrum Quality Products (Gardena, New Brunswick, USA). poly acrylic acid, Carbopol 934p from BF Goodrich Co. (Cleveland, Ohio, USA), anhydrous citric acid and sodium hydrogen carbonate from Winlab (Edgware, UKJ, magnesium stearate, glycerylmono stearate, and pepsin powder from BDH Chemicals Ltd. (Poole, England) were used as received. Commercial, 120 mg, extended-release verapamil tablets, each containing 120 mg verapamil as hydrochloride, lot No. M-lO, manufactured May 2000, expiry May 2003 (Knoll, Ludwigshafen, Germany). dihydroergotamine mesylate (Sandoz, East Hanover, NI. USA) and empty hard gelatin capsules (Feton, Coni-Snap~, Bruxelles, Belgium) were obtained from National Guard Hospital.

2.2.1.3. Preparation of tablets

Granules containing 1:1,2:1, and 1:2 ofVP:HPMC-4000 and 5 % effervescent mixture were compressed into tablets using a single-punch tableting machine (Erweka AR400) fitted with 9 mm flat-faced punches. Granulation was done using 96 % ethanol as granulating agent. A portion of granules from each formula containing an amount ofVP equivalent to 120 mg was weighed and fed manually to the die and compressed under the same conditions of die depth and compression load. 50 tablets were prepared from each formula.

2.2. Methods 2.2.1. Preparation of floating dosage forms

Aiming to prepare a sustained release floating systems for verapamil hydrochloride, the hydrocolloid polymers, HPMC-4000, HPC, EC and CP were employed. Floatation was achieved by adding an effervescent mixture of sodium bicarbonate and anhydrous citric acid. Some factors were investigated including the drug to polymer ratio, the granulating solution, coating granules with ethyl cellulose, and finally granule compression into tablets.

2.2.2. In vitro evaluation of the prepared floating dosage forms 2.2.2.1. Floatation characteristics
The test was performed by placing each of the capsule or tablet in a 250 ml beaker, containing 200 ml of simulated gastric fluid with pepsin USP-23, pH 1.2, maintained at 37 0.5C in a water bath. Their physical state was observed for 12 h. The time between introduction of the dosage form and its buoyancy on the simulated gastric fluid (Jag time). and the time during which the dosage form remains buoyant (duration of buoyancy) were determined visually. Three replicates of each formula were performed.

2.2.1.1. Preparation of verapamil capsules filled with powder or granulated-mix

The ingredients of each formula as described in Table 1 were mixed in a cubic mixer (Erweka KUl, Erweka Apparatebau, Heusenstamm, Germany) for 15 min and either filled directly into capsules or granulated manually using 80 % or 96 % alcohol by sieve granulation method where the wet mass was passed through 1.25 mm sieve followed by complete drying, sieving and mling into capsules. In each case a portion from each formula (passed through a sieve of pore size 1.25 mm and retained on a sieve of pore size 0.8 mm) containing VP equivalent to 120 mg was weighed and filled manually into hard gelatin capsules of suitable size.

2.2.2.2. Dissolution rate studies

The dissolution of VP from different formulations \va5 monitored using an automated dissolution monitoring system consisting of a USP dissolution apparatus No. 2 (SR8 Plus, dissolution test station, Hanson Research, USA) connected to an autosampler (Dissoette 2 autosampler model 27-6AS, Hanson Research). using a Lambda 11spectrophotometer (Perkin Elmer Instrument, Norwalk, CT) connected to an IBM computer and using Mastro 3.0 software from Hanson Research. Three capsules or tablets from each formula were tested. The dissolution medium \va5 900 ml of 0.1 N HCl (USP-23, pH 1.2). The stirring rate was 100 1 rpm and the temperature \va5 maintained at 37 0.5 0c. Five ml samples were withdrawn automatically at appropriate time intervals and replaced with fresh and preheated (37C) dissolution medium. Absorbance was measured spectrophotometrically at 278 nm. Preliminary experiments showed that the polymers did not interfere with the assay of the drug at this wavelength. The amount released at different time intervals \va5 calculated from the regression line of the standard curve developed in the same medium and the percentage drug release was computed for each formula based on the average drug content determinations.
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2.2.1.2. Preparation of ethylcellulose-coated granules

Aiming to investigate the effect of coating granules on sustaining the release of verapamil from the granulated matrices a portion of a formula containing 1:1 VP:HPMC was subjected to mm coating by EC using the pan coater (AR-4000, Erweka). The coating solution (l0 % w/v of EC in absolute ethanol) was sprayed over the granules in the coating pan and the solvent was allowed to evaporate after each spray using a hot-air blower. Four different coat percent (l0.8, 20.4, 33.24 and 40 %) were prepared by varying the application time and determined based on the overall weight gain afrer complete drying. Portions from each coated granule formula equivalent to 120 mg VP were weighed and filled manually into hard gelatin capsules of suitable size.

1q hh

"I~hp<hpnPI "I - Veranamil hvdrochloride

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"~ ....

,~~'~ .

2.2.3. In vivo evaluation of a selected floating formula 2.2.3.1. Evaluation of gastric retention using X-ray imaging
The selected tablet formula for in vivo investigation was reformulated with 12 % BaS04 as opaqueing agent and prepared by wet granulation with 96 % alcohol followed by compression (ErwekaAR400 tableting machine, fitted with 9 mm flat-faced punches). A control tablet, devoid of HPMC and the gas generating mixture, and containing Avicel PH 101 in addition to the dmg was prepared using the same proportion of BaS04' The X-ray studies were carried out using 6 male beagle dogs having weight range of 11.13 to 12.5 kg. In each experiment, the animals were allowed to fast overnight with free access to water and a radiograph was made just before the administration of the tablet to ensure the absence of radio-opaque material in the stomach. The two formulations (test and contro!) were administered by natural swallowing each to a group of three dogs followed by 50 ml of water. The radiographic imaging was taken to animals in a standing position and the distance between the source of X-rays and the animal was kept constant for all imaging, thus the observation of the tablet movements could be easily noticed. Radiographic imaging of the abdomen was taken using an X-ray machine (Model No. 3064581-B-531OJ. connected to a video cassette recorder SVO9500MD and a video graphic printer UP960-EC (Siemens, Germany) every 30-min intervals for a period of 6 h.

From the data of plasma concentration, the maximum plasma concentration (Cm"", ng/m!) and the corresponding time (tma.vh) were directly extracted for the two treatments in each individual animal. A plot of the mean plasma concentration versus time has been constructed for each treatment. The area under the plasma concentration-time curve from time zero to 24 h (AUCo-24h'ng . hIm!) was obtained by applying the trapezoidal mle. The AUC24_ was estimated by adding the area under the tail to AUCO-24h' The area under the tail was calculated by dividing the last measurable concentration by the elimination rate constant obtained by linear regression of the elimination phase of the plasma concentration versus time curve. The mean residence time (MRT, hJ. which is a non-compartmental pharmacokinetic parameter was obtained using the suitable equation [20] after the calculation of the area under the first-moment curve (AUMCo.~, ng . h2/m!). The relative bioavailabiliry (Frol) of the tested formula compared with the reference product was calculated as: Frol = [AUCo.~(tested formula) / AUCo.~ (reference product)] . lOO The significance of the difference between the two .treatments was evaluated by one-way analysis of variance (AI\lOVA)using Duncan's multiple comparison test on the computer program, Stat lOO, version 1.24, Biosoft (1996). Differences were considered significant at p $ 0.05.

3. Results and discussion

Anhydrous citric acid (CA) and sodium hydrogen carbonate in the ratio of 1:1 (w/w) and a concentration of 5 % per formula (based on preliminary experiments) were used as gas generating mixture (GG) during the course of this work. Ethanol 80 % (v/v) was used as a granulating agent for all the prepared granules as it produced more firm granules. However, granules meant to be compressed into tablets were prepared with 96 % ethanol as they were more compressible. 3.1. In vitro evaluation 3.1.1. Effect of the polymer type and the drug to polymer ratios The capsulation of a well-blended powder of different types of polymers (HPMC-4000, HPC. and EC) and different drug to polymer ratios was investigated. Only HPMC-4000 formulations combined a good floating and reasonable delay in drug release (Fig. 1). The three formulations containing 1:1 (Ft), 2:1 (F4), and 1:2 (Fs) of VP:HPMC showed exceIlent floating properties as they remained buoyant for more than 12 h. The formulae containing HPC (Fz) and EC (F3) showed fast release profile with 82.9 0.38 % and 86.80 2.43 % of the dose being released within 30 min, respectively. They both exhibited immediate floatation, however only the formulae containing EC remained floating during the course of the experiment (12 h). while that containing HPC sunk after 15 min only. The incorporation of CP was thought to improve the sustaining profile of VP from the powder-fiIled capsules containing HPC as a hydrophilic polymer. The results

2.2.3.2. Comparative 2.2.3.2.1. Treatment

bioavailabiIity

study

protocol and sample analysis

The selected floating formula containing 120 mg of VP was compared with a commercial sustained release VP product, which contains 120 mg VP. Five healthy beagle dogs (3 males and 2 females), having a body weight range of 9.5-12 kg, were used in this study. The single dose randomized design was applied where all dogs received the same treatment at the same time and a washout period of at least two weeks was allowed between treatments. The sequence of the trials was the standard commercial tablets followed by the test formula. The dogs were allowed to fast over night prior to and 4 h after administration of each treatment; thereafter, they resumed a normal unrestricted diet. They were placed in normal cages during the course of the experiment without using a restrainer. The treatments were given by normal swallowing followed by 50 ml water. 5 ml blood samples were collected, in heparinized evacuated glass tubes through an indwelling polyethylene cannula into the cephalic vein, at 0, I, 2, 3, 4, 5, 6, 9, 12, and 24 h from the time of administration. The plasma was immediately separated by aspiration after centrifugation at 3000 rpm for 5 min and frozen at -20 QC until analyzed. Animal 4 was excluded from the second round (floating tablet) due to its low health condition. The concentration of VP in the test samples was determined using the HPLC method introduced by Niazy and Jun [19].

2.2.3.2.2. Calculation and statistical of pharmacokinetic parameters

treatment

The pharmacokinetic parameters were calculated from the plasma level data obtained for the individual dogs and presented as mean SO.

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CD

0 40 ~ 50 20 30 0 10 60 70 80 90

'" 100
<J) CD

release from simple diffusion to concentration independent (zero order) by increasing the ratio of the polymer to the drug, it is obvious that increasing this ratio retarded the release rate of the drug significantly (Fig. 1). The tsO%for tlle formula containing 2:1 drug:HPMC was 2.17 0.14 h compared to 5.53 0.31 hand 7.95 0.19 h for the formulations containing 1:1 and 1:2 of VP:HPMC, respectively. A significant difference (p $ 0.05) was detected using one-way ANOVAbetween the three formulations.
__ ___ F1 (1:1. VP:HPMC-4(00) F2 (1:1. VP:HPC) VP:EC)

3.1.2. Effect of granulation

The granulation of two of the formulations that showed perfect performance in powder form concerning the sustained release and long floating period (F1 and Fs) lead to complete loss of the sustained release pattern (TSO% of 45 and 30 min, respectively) and the floating duration (1.5 and 0.25 h, respectively). Trying to retard the release after granulation, magnesium stearate, glyceryl mono stearate and CP were incorporated in 10 to 15 % per formula. These additives only improved tlle floating properties (3.5 to 12 h duration), while the release rate was even mildly accelerated (tSO% of 15 to 30 min). The slow release of the drug from the powder-filled capsules compared to the granule-filled capsules of the same composition was due to the congealing of the externallayer of the powder during the dissolution of the capsule shells in the former case. The congealed layer acted as a barrier for farther penetration of the dissolution medium or the diffusion of tlle drug. While, granules spread all over the dissolution medium once the gelatin shell dissolved exposing greater surface area in contact with the dissolution medium. Further more, being soluble in alcohol, which is used as granulating agent, VP concentrates more at the surface of granules upon evaporation of alcohol leading to further increase in the dissolution rate. Moreover, the granulating agent (ethanol) allowed the formation of porous structures in the granule formulations through which the dissolution medium can diffuse rapidly to extract the drug.

-.!o- F3 (1 :1.
-*-F4 -ill__ -+-F7

(2:1. VP:HPMC-4(00) FS (1 :2. VP:HPMC-4(00) F6 (1: 0.8: 0.2. VP:HPC:CP) (1: 0.6: aA, VP:HPC:CP)

10

15
Time (hours)

20

25

30

Fig. 1: Release profile of verapamil hydrochloride from powderfilled capsules formulated with different polymers and drug to polymer ratios.

showed that CP did not affect the release characteristics of the drug even upon the addition of 40 % CP per total polymer content (F7). None of the tested formulations showed any retardation to the release and the floating characteristics were poor as well. The release kinetic data (Table 2) revealed that the release mechanism from Fl (1:1 of VP:HPMC) and Fs (1:2 of VP:HPMC) followed zero-order kinetics that was confirmed by their release exponent (n) which de'scribes the mechanism of drug release as introduced by Korsmeyer et aI. [21]. However, F4 (2:1 ofVP:HPMC) followed Higuchi diffusion model as indicated by the highest correlation coefficient for the diffusion model and the closeness of the n value to 0.5 (r = 0.999, n = 0.504). Besides changing the mechanism of drug

Table 2: Kinetic data of the dissolution

rate of verapamll HCI from different formulations

prepared

by various techniques.

0.999 0.992 0.998 0.986 0.899 0.939 0.964 0.932 1.011 0.433 15.769 -1.989 8.021 0.928 0.932 0.679 0.504 -2.024 6.379 0.922 10.724 0.956 0.997 10.923 -1.133 -1.207 22.493 0.961 19.465 -4.184 6.495 6.799 -4.295 -1.252 0.741 0.672 0.704 0.994 0.973 0.995 0.999 33.627 -0.066 0.953 0.993 0.990 0.981 r: regression coefficient of the linear regression of different kinetic model equations.
FI (powder)

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100

90 80
70

.,60 VI

., 50
tU

-+-F1

(10.8% EC)
EC) EC)

~ 40 30 20

___ F1 (20.4% EC)

--e-

F1 (33.2%

-*" F1 (40.0%

10 o
o

10

15 Time (hours)

20

25

30

Fig. 2: Release proftle of verapamil hydrochloride from capsules filled with granules coated with different ethylcellulose coat percent.

3.1.3. Effect of coating granules with EC Coating the granules of the formula containing 1:1 of verapamil:HPMC with different percentage of EC (10.8, 20.4, 33.2,40.0) improved the sustaining of drug release; the higher the coat percent the slower the drug release attained (Fig. 2). The kinetic model of drug release was found to follow zero-order kinetics with n values of 1.01, 0.96, 0.93 and 1.01 for the four coat percentages. The statistical analysis of the tsO%of these formulations showed significant difference between the different formulations (P :s 0.05) except benveen the formulations having 32.2 and 40.8 % coat. All the coated granule formulations exhibited poor floating with only 5 min duration of buoyancy. This may be attributed to the hydrophobic nature of EC as the result of lacking any of the hydrophilic groups (hydroxyl or carboxyl), which hindered the hydration of the granules and hence their swelling that reduces their density. It is conclusive that EC coating, however controlled the release rate, was not suitable for floating dosage forms due to its slow water permeation and lack of flexibility. These results are in agreement with what Kr6gel and Bodmeier [13] have reported.

tively. This could be attributed to the slow rate of water uptake (Iow rate of expansion) with increasing the percentage of polymer per tablet. The % increase in the tablet weigh after 15 min was determined for each formula and found to be 69.90 3.57, 63.07 1.30, and 60.27 1.22 for F4' F I and Fs, respectively. The rate of water uptake is essential for the absorption of dilute hydrochloric acid solution, followed by gas generation and floatation, this is beside the reduction of density that follow expansion. The release rate was dependent on the polymer ratio as well. The percent amounts released after 6 h were 55.12 0.52,46.24 1.90, and 33.66 1.58 % for F4, F 1> and Fs, respectively (Fig. 3). Tableting the granules led to much slower release rate than filling them directly into capsules. They also showed slower release rate than the powder-filled capsules ofthe same drug:HPMC ratios. Granules compression allowed the formation of a cohesive gel layer after wetting of the tablet surface. The rate of water diffusion through this layer was critical for the release rate of VP.The capsulated powder of the same drug:polymer ratio absorbed more water and so the formed gel layers were less firm and integer than those formed with the compressed tablets leading to faster release rate. The kinetic treatment of the dissolution data of tablets revealed that the release mechanism of VP from the tableted granules followed non-Fickian diffusion model based on their n values (Table 2). The statistical analysis of the tsO%calculated from the diffusion equation for the tableted formulations revealed the presence of significant differences benveen the three formulations.

100

80

j .,
VI

.,60

a:
g<

40
___ F1(1:1 VP:HPWC-4000)

--e- F4(2:1 VP:HPWC-4000) 3.1.4. Effect of tableting the granules

20 ~ F5(1:2 VP:HPWC-4000)

The objectives of the present study (good floating properties and sustaining drug release) were achieved by tableting the prepared granules of FJ, F4' and Fs containing different VP:HPMC ratios of 1:1, 2:1 and 1:2, respectively. All of the compressed granules showed buoyancy during the entire time course of the experiment (12 h). Only F4 showed immediate flotation while F1 and F5 showed a lag period of 4 and 18 min, respecPhann. Ind. 66, Nr. 11, 1364-1372 (2004)
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o
o

10

15
Time (hours)

20

25

30

Fig. 3: Release proftle of verapamil hydrochloride from tablets of granules prepared with different verapamil:HPMC ratios.

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30

min

1 h

2h

Fig. 4: X-ray photographs taken after the administration of control tablets to beagle dogs.

3.2. In vivo evaluation Based on the in vitro study a formula of verapamil to HPMC (120 mg each), ating mixture and prepared by wet 96 % alcohol followed by compression further in vivo evaluation.
3.2.1. Gastric retention

composed of 1:1
5 % of gas gener-

granulation with was selected for

istered in the fasted state is the third phase of the migrating myoelectric complex (MMC), which occurs approximately every 2 h in humans [25] and approximately every 1 h in dogs [26]. For this phase the intense activity that empties large, non-disintegrating particles including dosage forms from the stomach to the small intestine is characteristic. As the results have shown that the mean gastric residence time for the developed floating tablets was more than 4 h in dogs, this means that the floating formula withstands four of these phases without empting. As the comparison of the strength of gastric motility and stomach emptying between humans and dogs shows no big differences [25, 26] it is expected that the gastric residence of this formulation in human beings would be even longer.
3.2.2. Comparative

period

Many authors questioned the gastric retention capabilities of floating systems [22, 23]. They concluded that the mean determinant of emptying appeared to be the food intake and not the density of the dosage form. It was decided here to undergo X-ray imaging experiments on a fasted dog in order to abolish the effect of food and gastric content on emptying. The behavior of the tablet in the dog stomach was observed using a radiographic imaging technique. Selected samples from radiographic images of dogs' abdomens after the administration of control tablets and floating tablets are shown in Fig. 4 and 5, respectively. The control tablets were seen in the stomach during the first hour (only during two inspection periods, each is 30 min interval) in two of the three dogs. The tablet disappeared from the second radiogram image of the third dog indicating less than 1 h emptying. This was expected as the control formula lacks any hydrogel-forming polymer and may disintegrate rapidly. On radiographic images made 1 h after the administration of the floating tablets, they were observed in the animal's stomach (Fig. 5). In the picture taken at 2 h, significant changes in positions were detected, which provide evidence that the tablets did not adhere to the gastric mucosa but, on the contrary, floated on the gastric fluid. This result is in agreement with the finding of Baumgartner et al. [24]. The tablets disappeared from the stomach images taken after 5 h for the three tested dogs. This shows that the gastric emptying time could be more than 4 h and less than 5 h. The major limitation to the upper gastrointestinal residence time of solid single unit dosage forms admin-

bioavailability

of verapamil

Fig. 6 shows the change in plasma concentration of VP with time after oral administration of the reference standard, and the floating tablet to beagle dogs. Table 3 includes the pharmacokinetic parameters ( SO) generated from the analysis of the individual data. The Cmax was found to be 51.35 15.53 and 50.59 12.33 ng/ml for the reference standard and floating tablets, respectively, and the corresponding tmax were 7.00 1.87 and 9.5 3.32 h. It is obvious that the values of Cmax were very close for the two treatments and no significant difference was obtained between them. The floating tablets exhibited delayed tmax' however, non-significantly different from the standard product at p < 0.05. Both the data of Cmax and tmax values were comparable with reported values in the literature [27]. They reported tmax of 6.7 and 8.3 hand Cmax of 122.7 58.1 and 93.8 38.7 ng/ml for two sustained release 240 mg VP capsule and tablet, respectively, administered to human volunteers. The tl/2 were 11.022 3.818 and 11.444 3.542 h for standard product, and the floating formula, respectively, which were not significantly different. Muirhead et al. [27] reported tl/2 of 9.6 and 9.2 h for the sustained

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1h

2h

3h

4h

Fig. 5: X-ray photographs taken after the administration of the floating tablets to beagle dogs.

VP capsule and tablet, respectively. A t1/2of 10.68 h was reported after administration of floating pellet to human volunteers [281. The values of the MRT, which is the non-compartmental analogue of t1/2' were also parallel to those of t1/2. The tested formula showed a slightly higher MRT (15.34 1.56 h) than the reference standard (14.70 1.72 h), however, there was no statistical difference between them. No statistical difference was also observed in the
AUCo_", for the reference standard (949.84 245.11 ng . hI ml) and the floating tablet (1111.60 433.93 ng .

betvveen the floating formula and the reference standard can be simply concluded based on the AUCo_", and Cmax values that lie within the acceptance range of the
FDA (80 - 125 %).

h/ml) , however the later showed higher one. The relative bioavailability was 117.03 45.68 %. Bioequivalence

70

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0,
.Srn

60

2
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Q)

-e- Referencetablet ___ Floatingtablet

50

~ c
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(J
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Q) tU

40

~ 30
:>

E 20

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tU Q)

VP is considered an example of drugs with high pharmacokinetic variability. Tsang et al. [29] reported a very high intra-subject variability with VP. They concluded that the high variability complicates bioequivalence assessment and can lead to an erroneous assumption. However, tlle pharmacokinetic data presented here did not show extraordinary variability when compared with most published bioavailability studies. The maximum RSD % was 39.04 %. The floating tablet showed more sustained release characteristics compared with the reference standard, although failed to show significant difference. As a conclusion, it was obvious that the floating formula was able to delay the gastric emptying ofVP tablet in beagle dogs. Knowing that the dogs undergo faster migrating myoelectric complexes than humans, the significant delay in the gastric emptying observed with the floating tablets (> 4 h in dogs) when compared to the control (less than 1.5 h) is expected to be even longer in humans. This would maximize absorption by allowing the slowly released drug in the stomach to reach the upper small intestine (the sight of absorption) in a form ready for absorption. The floating tablets showed bioequivalence with a commercial sustained release tablet containing 120 mg verapamil with higher mean AUCo_", and Cmax and longer tmax, however, non-significantly different.

~
10

4. References
5

10

15
Time (hours)

20

25

30

Fig. 6: Mean plasma verapamll HCI concentrations after a single oral dose of the reference standard tablets and floating tablets to beagle dogs.
Phann. Ind. 66, Nr. 11, 1364-1372 (2004)

[1] McTavish, D., Sorkin, E. M., Verapamil. An updated review of its pharmacodynamic and pharmacokinetic properties, and therapeutic use in hypertension, Drugs 38, 19 (1989) [2] Klein, H. A., KapIinsky, E., Digitalis and verapamil in atrial fibrillation and flutter. Is VP now the preferred agent? Drugs 31, 185 (1986)

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Plkheshen et al. - VeraDamil hvdrochloride

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Table 3: Pharmacokinetlc parameters of verapamll (mean SD) following a single oral dose administration standard tablet and the "oatlng tablets to beagle dogs under fasting conditions. 117.03 12.3 24.4 50.6 11.72 15.34 756.10 29.70 26.7 34.9 30.2 i5.5 17465.80 35.400 30.746 10.15 0.020 0.025 8651.33 1.56 4246.79 1.72 30.30 0.065 49.53 34.639 8933.17 1111.60 30.955 3110.61 224.53 2482.63 204.22 245.11 29.61 25.80 39.04 433.93 45.68 3.542 11.444 3.818 31.96 39.04 34.82 3.3 1.9 9.5 CV%

of the reference

7.0 51.4 14.70 7767.94 689.61 949.84 11.022 14013.56 0.070

SD

[3] Jankowski, A, Marzec, A, Lamparczyk, H., Comparative bioavailability of verapamil from rapidly absorbed and slow release preparations. J. Pharm. Biomed. Anal. 10, 1101 (1992) [4] Goracinova, K, Klisarova, L., Simov, A. et al., Physicochemical stability of controlled-release solid dispersion granules containing verapamil HCl as active substance. Acta Pharm. 48, 93 (1998) [5] Moes, A. J., Gastroretentive dosage forms. Crit. Rev. Ther. Drug Carrier Syst. 10, 143 (1993) [6] Sheth, P. R., Tossounian, J. L., The hydrodynamically balanced system (HBSTM): A novel drug delivery system for oral use. Drug Dev. Ind. Pharm. 10,313 (1984) [7] Gerogiannis, V.S., Rekkas, D. M., Dallas, P.P. et aI., floating and swelling characteristics of various excipients used in controlled release technology. Drug Dev. Ind. Pharm. 19, 1061 (1993) [8] Michaels, A. S., Bashaw, J. D., Zaffaroni, A., Gastro-inflatable drug delivery device, US Pat. 3,901,232 (1975) [9] Watanabe, S., Kayano, M., Ishino, Y., et al., Solid therapeutic preparation remaining in the stomach, US Pat. 3,976,764 (1976) [10J Harrigan, R. M., Intra-gastric floating drug delivery device, US Pat. 4,055,178 (1977) [11] Ushimaru, K, Nakamichi, K, Saito, H., Slow release preparation, UK Pat. Appl. GB,2,163,648,A,1-8 (1985) [12] Ichikawa, M., Watanabe, S., Miyake, Y.,A new multipleunit oral floating dosage system. I: Preparation and in-vitro evaluation of floating and sustained-release characteristics. J. Pharm. Sci. 80, 1062 (1991) [13] Krogel, I., Bodmeier, R., Floating or pulsatile drug delivery systems based on coated effervescent cores. Int. J. Pharm. 187, 175 (1999) [14] Babu, V. B. M., Khar, R. K., In-vitro and in-vivo studies of sustained-release floating dosage forms containing salbutamol suifate. Pharmazie 45, 268 (1990) [15] Khattar, H., Ahuja, A., Khar, R. K., Hydrodynamically balanced systems as sustained release dosage forms for propranolol hydrochloride. Pharmazie 45, 356 (1990) [16] Mazer, N., Abisch, E., Gfeller, J. et al., Intra-gastric behavior and absorption kinetics of a normal and floating modified-release capsule of isradipine under fasted and fed conditions. J. Pharm. Sci. 77, 647 (1988) [17] Chen, G., Hao, W., In-vitro performance of floating sustained-release capsule ofverapamil. Drug Dev. Ind. Pharm. 24, 1067 (1998) [18] Rouge, N., Cole, E., Doelker, E. et al., Buoyancy and drug release patterns of floating minitablets containing piretanide and atenolol as model drugs. Pharm. Dev. Technol. 3, 73 (1998)

[19] Niazy, E. M., Jun, H. W., Rapid quantitation ofverapamil in plasma by high performance liquid chromatography. Anal. Lett. 18, 1103 (1985) [20J Gibaldi, M., Perrier, D., Pharmacokinetics, 2nd ed., pp. 149-152, Marcel Dekker, New York (1982) [21] Korsmeyer, R. w., Gurny, R., Doelker, E. et al., Mechanisms of solute release from porous hydrophilic polymers. Int. J. Pharm. 15,25 (1983) [22] Mtiller-Lissner, S. A., Blum, A. L., The effect of specific gravity and eating on gastric emptying of slow-release capsules. New Engl. J. Med. 304, 1365 (1981) [23] Davis, S. S., Stockwell, A. E, Taylor, M. J. et aI., The effect of density on the gastric emptying of single-and multiple-unit dosage forms. Pharm. Res. 3, 208 (1986) [24] Baumgartner, S., Kristl, J., Vrecer, E et aI., Optimization of floating matrix tablets and evaluation of their gastric residence time. Int. J. Pharm. 195, 125 (2000) [25] Hwang, S. J., Park, H., Park, K., Gastric retentive drugdelivery systems. Crit. Rev. Ther. Drug 15, 243 (1998) [26] Cunningham, J. G., in: Text book of veterinary physiology, 2nd ed., Cunningham, J. G. (ed.), p. 272. Saunders, Philadelphia (1997) [27] Muirhead, 0. C., Norris, R. J., Cockayne, D. et al., Pharmacokinetics of once-daily verapamil: Comparative bioavailability of two sustained-release formulations. Brit. J. Clin. Pract., Suppl. 60, 14 (1988) [28] Sawicki, W., Pharmacokinetics ofverapamil and norverapamil from controlled release floating pellets in humans. Eur. I. Pharm. Biophann. 53, 29 (2002) [29] Tsang, Y. c., Pop, R., Gordon, P. et al., High variability in drug pharmacokinetics complicates determination of bioequivalence: experience with verapamil. Pharm. Res. 13, 846 (1996)

Correspondence:
Seham A. Elkheshen, Faculty of Pharmacy, Cairo University, Kasr EI-Eini Street, Cairo 11562 (Egypt), e-mail: Selkshen@hotrnail.com
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