Journal of Current Pharmaceutical Research 2010; 01: 31-38

JCPR 2010; 01: 31-38 2010 Medipoeia Received: 15/03/2010 Revised: 21/03/2010 Accepted: 30/03/2010

Development and evaluation of buccoadhesive drug delivery system for Atorvastatin calcium
Asha S. John, Sathesh B. P. R, Goli Divakar, Manoj K. Jangid and Kapil K. Purohit

Asha S. John, Sathesh B. P. R, Goli Divakar, Manoj K. Jangid & Kapil K. Purohit Department of Pharmaceutics, Acharya & B.M Reddy college of Pharmacy, Soladevanahalli, Chikkabanavara (Post), Hesaraghatta Main Road, Bangalore-90.

ABSTRACT Objective: The purpose of this research was to study mucoadhesive bilayer buccal tablets of Atorvastatin calcium using the bioadhesive polymers Carbopol 934P (CP), Sodium CMC, Hydroxy ethyl cellulose (HEC) and Sodium alginate (Na-alginate) along with ethyl cellulose as an impermeable backing layer. Materials & Methods: Nine different formulations of mucoadhesive buccal tablets of Atorvastatin calcium were prepared, which contain the polymers in various combinations. Tablets were prepared by direct compression method and characterized by swelling studies, % matrix erosion, surface pH, bioadhesive properties, in-vitro drug dissolution and in-vitro diffusion studies. Result: All the formulations gave the satisfactory results in terms of bioadhesive performance, physical and mechanical properties and surface pH. The swelling index was proportional to CP content and inversely proportional to sodium CMC content. The surface pH of all tablets was found to be satisfactory, close to neutral pH; hence, buccal cavity irritation should not occur with these tablets. Drug release and drug diffusion from the tablets were depended on the ratio and type of the polymer used in the formulation. Tablets containing CP and Na-CMC in the ratio of 3:2 (F2) had the maximum percentage of in-vitro drug release without disintegration in 6 h. The mechanism of drug release was found to be non-Fickians diffusion and followed anomalous release. The formulation F2 was optimized based on good bioadhesive strength (19.0 0.30 g) and sustained in vitro drug permeation (85.68 % for 6 h). The chosen tablet containing 8 mg of Atorvastatin calcium performed 6 h sustained drug release with desired therapeutic concentration. Conclusion: The formulation F2 was optimized based on good bioadhesive strength (19.0 0.30 g) and sustained in vitro drug permeation (85.68 % for 6 h). The chosen tablet containing 8 mg of Atorvastatin calcium performed 6 h sustained drug release with desired therapeutic concentration. Keywords: Mucoadhesion; bilayer device; buccal drug delivery; CMC sodium; PEG 6000; MCC; HEC; Atorvastatin calcium.

1. INTRODUCTION The buccal mucosa has been investigated for local and systemic delivery of therapeutic peptides and other drugs that are subjected to first-pass metabolism or are unstable within the rest of the gastrointestinal tract.1 Buccal delivery offers a safer mode of drug utilization, since drug absorption can be promptly terminated in cases of toxicity by removing the dosage form from the buccal cavity (Nakhat et. al., 2007). A suitable buccal drug delivery system should possess good bioadhesive properties, so that it can be retained in the oral cavity for the desired duration. In addition, it should release the drug in a unidirectional way toward the mucosa, in a controlled and predictable manner, to elicit the required therapeutic response. This unidirectional drug release can be a chieved using bilayer devices (Vishnu et. al, 2007). Atorvastatin calcium is HMG Co - A reductase widely used in the treatment of hyperlipidemias and cardiovascular diseases and it

Correspondence: Asha S. John Department of Pharmaceutics, Acharya & B.M Reddy college of Pharmacy, Soladevanahalli, Chikkabanavara (Post), Hesaraghatta Main Road, Bangalore-90. E-mail: ashasusanj@gmail.com

Journal of Current Pharmaceutical Research 2010; 01: 31-38

is known to have low oral bioavailability (14%) due to an extensive high first-pass effect and its availability in less dose size i.e., in few mg. Hence, it was considered as suitable candidate for administration via buccal route. The aim of the present study was to design and develop buccoadhesive controlled release tablets of Atorvastatin calcium that could be applied to the buccal mucosa to release the drug unidirectionally in buccal cavity in order to decrease gastric irritation and avoid first pass effect for improvement in bioavailability, to reduce the dosing frequency and to improve patient compliance. 2. MATERIALS AND METHODS Material Atorvastatin calcium was obtained as a gift sample from Unichem Labs (Mumbai, India). Carbopol 934P and ethyl cellulose were procured from Central Drug House, New Delhi. Sodium CMC and MCC were obtained from NR Chem (Mumbai). Hydroxy ethyl cellulose was procured from Loba Chemie (Bangalore) and Sodium alginate from Thomas baker (Bangalore). All other reagents and materials were of analytical or pharmacopoeial grade. Preparation of buccoadhesive bilayered tablets of Atorvastatin calcium Drug can be incorporated in the adhesive layer of the tablet, which comes in contact with the mucosal surface. The drug containing mucoadhesive layer is then protected from the oral cavity environment by a second upper inert layer, which faces into the oral cavity. Buccoadhesive tablets were formulated by direct compression method. All the ingredients of the formulation were passed through a sieve # 60 and were blended in a mortar with a pestle to obtain uniform mixing. The blended powder of the core was compressed on a 9 mm punch in a single stroke multi station tablet punching machine. After punching the core layer, upper punch was removed and ethyl cellulose was added over it and again compressed. Tablets weighing 125 mg and hardness 4.5 kg/cm2 were obtained. Formulation chart is given in Table 1. All tablets contained 31% MCC as filler, 8% PEG as permeation enhancer, 3% magnesium stearate as lubricant, and 50% of total bioadhesive polymers with different mixing ratios of carbopol 934P and CMC sodium, carbopol 934P and HEC and carbopol 934P and sodium alginate with respect to 100 mg core of the tablet. Drug content Five tablets were accurately weighed and powdered. A quantity of the powder equivalent to 8.0 mg of Atorvastatin calcium was weighed accurately and extracted in 100 ml methanol by shaking for 20 min. After filtration through whatmann filter paper no.1 and sufficient dilution with methanol, samples were
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analyzed spectrophotometrically at 246 nm. This procedure was repeated thrice. Amount of drug present was determined from the standard curve of Atorvastatin calcium in methanol Swelling study Three buccal tablets were weighed individually (W1) and placed separately in 2% agar gel plates with the core facing the gel surface and incubated at 37 1C. After every 1 h time interval until 6 h, the tablet was removed from the petri dish and excess surface water was removed carefully with blotting paper. The swollen tablet was then reweighed (W2) and the swelling index (SI) were calculated using the formula given in equation (Vishnu et. al., 2007). Swelling Index = [(W2-W1) W1] 100 Where, W1 = initial weight of the tablet W2 = final weight of the tablet Matrix erosion After swelling study, the swollen tablets were dried at 60c for 24 h in an oven and kept in descicator for 48 h and reweighed (W3). Matrix erosion was calculated using following formula (Ramana MV. et. al., 2007). % Matrix erosion = [(W1-W3) W3] 100 Surface pH study The surface pH of the buccal tablets was determined in order to investigate the possibility of any side effects in-vivo. As the acidic or alkaline pH may cause irritation to the buccal mucosa, the pH was maintained to neutral as closely as possible. A combined glass electrode was used for this purpose. The tablet was allowed to swell by keeping it in contact with 1 ml of distilled water (pH 6.5 0.05) for 2 h at room temperature. The pH was measured by bringing the electrode in contact with the surface of the tablet and allowing it to equilibrate for 1 min. Bioadhesion time The ex-vivo mucoadhesion time was examined after application of the buccal tablet on freshly cut sheep buccal mucosa. The fresh sheep buccal mucosa was tied on the glass slide, and a mucoadhesive core side of each tablet was wetted with 1 drop of phosphate buffer pH 6.8 and pasted to the sheep buccal mucosa by applying a light force with a fingertip for 30 seconds. The glass slide was then put in the beaker, which was filled with 200 ml of the phosphate buffer pH 6.8 and kept at 37 1C. After 2 minutes, stirring was applied slowly to simulate the buccal cavity environment, and tablet adhesion was monitored for 7 h. The time for the tablet to detach from the sheep buccal mucosa was recorded as the mucoadhesion time (Vishnu et. al., 2007).

Journal of Current Pharmaceutical Research 2010; 01: 31-38

Table 1: Formulation chart of buccoadhesive tablets of Atorvastatin calcium

Ingredients (mg) Core layer Atorvastatin Calcium MCC Carbopol 934P CMC Sodium HEC Sodium Alginate PEG 6000 Magnesium Stearate Backing layer Ethyl cellulose Total weight 8 31 40 10 --8 3 8 31 30 20 --8 3 8 31 10 40 --8 3 8 31 40 -10 -8 3 8 31 30 -20 -8 3 8 31 10 -40 -8 3 8 31 40 --10 8 3 8 31 30 --20 8 3 8 31 10 --40 8 3 Batch I F1 F2 F3 Batch II F4 F5 F6 Batch III F7 F8 F9

and analyzed after appropriate dilution by UV spectrophotometer at 241 nm (Jafar et. al., 2004). In-vitro diffusion studies The in-vitro buccal drug permeation study of Atorvastatin calcium through the sheep buccal mucosa was performed using Franz diffusion cell at 37 0.2C. Fresh sheep buccal mucosa was mounted between the donor and receptor compartments. The buccal tablet was placed with the core facing the mucosa and both compartments were clamped together. The donor compartment was filled with 1 ml of phosphate buffer pH 6.8. The receptor compartment (55 ml capacity) was filled with isotonic phosphate buffer pH 7.4 and the hydrodynamics in the receptor compartment was maintained by stirring with a magnetic bead at 50 rpm. 1 ml samples were withdrawn at predetermined time intervals and after appropriate dilution with isotonic phosphate buffer pH 7.4, analyzed at 240.5 nm using a UV spectrophotometer (Jafar A. et. al., 2004). Kinetics modeling of drug dissolution profiles The dissolution profile of best formulation was fitted to Zero order, First order and Higuchi to ascertain the kinetic modeling of the drug release. The method was adopted for deciding the most appropriate model (Rao et. al. 2007, Brahmankar et. al., 2003). Stability studies The purpose of stability study is to provide evidence on the quality of a drug substance or drug product which varies with time under the influence of a variety of environmental factors such as temperature, humidity and light. One formulation was selected for stability studies on the basis of the in-vitro drug release profile. The formulation was subjected to accelerated stability studies as per ICH (The International Conference of Harmonization) guidelines. The most satisfactory formulation was sealed in an aluminum foil and stored at 30 2C, 65 5 % RH and at 40 2C, 75 5 % RH for 2 months. Tablets were periodically removed and evaluated for physical characteristics, mucoadhesive properties, in-vitro drug release and in-vitro diffusion studies. 3. RESULTS AND DISCUSSION The main aim of this work was to develop buccoadhesive bilayered tablets to release the drug at mucosal site in unidirectional pattern without wash out of drug by saliva. CP, Sodium CMC, HEC and Sodium alginate were selected as buccoadhesive polymers on the basis of their matrix forming properties and mucoadhesiveness while ethyl cellulose, being hydrophobic as backing membrane. At preformulation study for drug polymer compatibility by FTIR gave conformation about their purity and showed no interaction

25 125

25 125

25 125

25 125

25 125

25 125

25 125

25 125

25 125

Bioadhesive strength Bioadhesive strength of the buccal tablets was measured on the Modified Physical Balance method.The method used sheep buccal membrane as the model mucosal membrane. The fresh sheep buccal mucosa was cut into pieces and washed with phosphate buffer pH 6.8. A piece of mucosa was tied to the glass slide which was moistened with phosphate buffer pH 6.8. The tablet was stuck to the lower side of another glass slide with glue. The both pans were balanced by adding an appropriate weight on the left- hand pan. The glass slide with mucosa was placed with appropriate support, so that the tablet touches the mucosa. Previously weighed beaker was placed on the right hand pan and water (equivalent to weight) was added slowly to it until the tablet detach from the mucosal surface. The weight required to detach the tablet from the mucosal surface gave the bioadhesive strength. The experiment was performed in triplicate and average value was calculated (Ramana et. al., 2007). In-vitro dissolution studies The USP dissolution test apparatus (apparatus II paddle type) was used to study the drug release from the tablets. The dissolution medium was 500 ml of phosphate buffer pH 6.8. The release was performed at 37 0.5C, with a rotation speed of 50 rpm. The backing layer of buccal tablet was attached to the glass slide with instant adhesive (cyanoacrylate adhesive). The slide was allocated to the bottom of the dissolution vessel. 5 ml samples were withdrawn at predetermined time intervals and replaced with fresh medium. The samples were filtered through whatmann filter paper

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Journal of Current Pharmaceutical Research 2010; 01: 31-38

(a)

(b)

(c)

(d)

(e) Fig. 1. FT-IR spectra of pure drug Atorvastatin calcium (a), physical mixture of drug and carbopol 934P 90 (b), physical mixture of drug and sodium CMC (c), physical mixture of drug and HEC (d) and physical mixture of drug and sodium alginate (e).

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Journal of Current Pharmaceutical Research 2010; 01: 31-38

Table 2. Physicochemical parameters of developed buccoadhesive tablets of Atorvastatin calcium

Code

Diameter (cm)

Hardness (kg/cm2)

Thickness (mm)

Weight uniformity (mg)

Friability (% loss)

% Drug content

Surface pH

F1 F2 F3 F4 F5 F6 F7 F8 F9

0.923 0.002 0.921 0.002

4.7 0.45 4.6 0.56 4.8 0.25 4.9 0.37 4.3 0.47 4.7 0.32 4.8 0.55 4.9 0.20 4.7 0.32

2.19 0.01 2.19 0.02 2.19 0.03 2.20 0.03 2.21 0.03 2.19 0.03 2.20 0.02 2.19 0.04 2.21 0.03

125.23 1.59 125.02 1.61 125.57 1.84 125.43 1.79 125.16 1.82 125.06 1.46 125.43 1.79 125.21 1.63 125.54 1.72

0.43 0.43 0.43 0.44 0.42 0.47 0.41 0.43 0.47

97.87 0.81 97.85 0.47 97.55 1.23 97.46 0.52 96.46 1.46 97.00 1.69 97.55 1.60 96.79 1.30 97.87 1.74

6.01 0.22 6.43 0.27 6.17 0.20 6.17 0.19 6.57 0.33 6.19 0.30 6.29 0.30 5.91 0.49 6.32 0.38

0.925 0.003 0.922 0.002 0.923 0.003 0.923 0.003 0.922 0.002 0.925 0.001 0.924 0.002

between drug and selected polymers.[ Fig. 1. (a) - (e)] All the formulations were found to be satisfactory when evaluated for weight uniformity (125.29mg), content uniformity (97.38%), thickness (2.19mm), hardness (4.71kg/cm2) and friability (0.44%).Thus all tablets comply with the IP standards. The surface pH of all formulations was within a range of 5.91 to 6.57, close to neutral pH. These results reveal that all the formulations provide an acceptable pH in the range of salivary pH (5.5 to 7.0). They did not produce any local irritation to the mucosal route. The results of all the above mentioned tests are shown in [Table-2]. The bioadhesion and drug release profile are dependent upon swelling behavior of the tablets. Swelling index increased as the weight gain by the tablets increased proportionally with the rate of hydration. In swelling study, it was found that the amount of carbopol plays an important role in swelling of the matrix and leads to the drug diffusion. The fastest hydration rate was obtained from batch I (sodium CMC) and batch II (sodium alginate) that hydrated above 50 % within the 3rd h. It was observed that swelling rate increased with an increase in CP content of the prepared tablets. Maximum swelling was seen in the formulations F1, F2 and F7. The mucoadhesive polymers used were hygroscopic and retain large amounts of water. Tablets containing sodium CMC (batch I) and sodium alginate (batch III) as secondary polymers
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were found to exhibit least matrix erosion. This is because the presence of water that balanced the weight loss due to erosion and was more evident for this groups, as they showed higher hydration rates. The plots of percentage swelling index and matrix erosion are shown in [Fig. 2. & 3].

Fig. 2. Percentage swelling of developed buccal tablets The ex-vivo mucoadhesive properties of the tablets were determined using sheep buccal mucosa. Tablets containing a higher proportion of CP showed higher mucoadhesion. The reason might be ionization of CP at salivary pH and formation of secondary bonds with mucin because of rapid swelling and interpenetration of the polymer chains in the interfacial region, which leads to

Journal of Current Pharmaceutical Research 2010; 01: 31-38

presence of higher amount of carbopol which will ionizes at pH environment of the dissolution medium. Ionization of CP leads to the development of negative charges along the backbone of the polymer. Repulsion of like charges uncoils the polymer into an extended structure. The counter ion diffusion inside the gel creates an additional osmotic pressure difference across the gel leading to

Fig. 3. Matrix erosion of developed buccal tablets improved attachment of the device to mucosal surface, while other polymer undergo superficial bioadhesion only. As the amount of sodium CMC increases adhesion force decreases, which might be due to the low viscosity of sodium CMC. This indicates that the bioadhesive strength of CP is much more than sodium CMC. The bioadhesive properties exhibited by the sodium CMC and sodium alginate tablets can be considered satisfactory for maintaining them in the oral cavity. The plots of bioadhesion characters against time are shown in [Table-3]. Table 2. Bioadhesive properties of prepared buccoadhesive tablets
Formulation F1 F2 F3 F4 F5 F6 F7 F8 F9 Bioadhesion time* (h) 7.18 0.09 6.37 0.07 6.18 0.13 6.39 0.16 6.20 0.09 6.08 0.36 6.39 0.06 6.20 0.05 6.08 0.36 Bioadhesion strength* (g) 21.6 0.67 19.0 0.30 17.2 0.36 16.9 0.80 14.2 0.55 12.2 0.30 19.7 0.40 18.2 0.41 16.1 0.20

(a)

(b)

Release of drug from the buccal mucoadhesive tablets varied according to the type and ratio of matrix-forming polymer. Carbopol 934P has excellent mucoadhesive, gelling properties and also helps in sustaining effect. Carbopol is more hydrophilic. It can swell rapidly, therefore decrease of carbopol content delays the drug release from tablet core. The maximum cumulative percent of drug release from formulation F1 could be attributed to the
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(c) Fig. 4. Dissolution profiles of batch I (a), batch II (b) and batch III (c)

Journal of Current Pharmaceutical Research 2010; 01: 31-38

the considerable swelling of the polymer. The continued swelling of polymer matrix causes the drug to diffuse out from the formulation at a faster rate. In theory, the higher the uptake of water by polymer, the more the amount of drug diffused out from the polymer matrix. Thus, this high amount of water uptake by polymers may lead to considerable swelling of polymer matrix, allowing drug to diffuse out at faster rate. Tablets from the formulation F1 (sodium CMC) showed a higher percentage of drug release compared to other groups. But due to the high matrix erosion of tablet, it cant be used for the further studies and formulation F2 was considered as it has got least erosion and good release character. The plots of percentage cumulative drug release against time are shown in Fig. 4. Formulation F2 was selected for further studies based on in-vitro drug release (93.75%), swelling index (93.67%), matrix erosion (16.90%), bioadhesive time (6.37h) and bioadhesive strength (19.0g).Kinetic treatment (Peppas et.al) of obtained result from in-vitro drug release data represents the anomalous release mechanism. The values of n and k are inversely related. A higher value may suggest burst drug release from the matrix. According to the criteria for release kinetics, a value of release exponent, n = 0.45, 0.45< n < 0.89, and 0.89< n <1.0 indicates Fickian (case-I), non-fickian (anomalous) and zero order (case-II) transport, respectively. The obtained values of n (diffusion exponent) was found 0.82 which lie between 0.45 and 0.89 in the formulation F2 exhibiting a non- Fickians release behavior and showed anomalous release. Analysis of drug release mechanism are shown in [Table4] Table 4: Correlation coefficients of drug release curves from the most satisfactory formulations of buccoadhesive tablets on kinetic models
Kinetic models Higuchi equation Peppas equation 1st order equation Zero order equation Mechanism N 30.74 0.82 -0.12 16.05 R2 0.77 -0.88 0.98

further studies and formulation F2 can be considered. The % drug released from this group was found to be 93.75 %.

Fig. 5. In-vitro diffusion profile of formulations F2, F4 and F7 buccal tablets.

Fig. 6. In-vitro diffusion profile of formulation F2 during stability studies x- Diffusion profile of F2 before stability study.

Anomalous

Since, the permeability of oral mucosa is low; the oral mucosal route could be utilized for potent drugs, which are required in small doses. Formulation F2 was subjected to in-vitro permeation study using diffusion cell. The results showed drug permeation of 85.68%. Tablets from the formulation F1 (sodium CMC) showed a high percentage of drug release compared to other groups. But due to the erosion of tablet, it cant be used for the
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This is because of higher percentage of hydration. From the results it was found that formulation F2 can be considered as the most satisfactory formulation because of its good adhesive properties, least matrix erosion and good release behavior and have chosen for the stability studies. The plots of percentage cumulative drug diffused against time are shown in Fig. 5. After the 2 months storage of formulation F2, values of all parameters like diameter, thickness, % drug content, friability, surface pH, bioadhesive properties, swelling index and % matrix erosion were checked periodically and found to be almost similar to the initial values. The drug dissolution and diffusion profile

Journal of Current Pharmaceutical Research 2010; 01: 31-38

were similar to the initial profile (Fig. 6). There was not any significant change in any value. So it can be said that formulation is stable. 4. CONCLUSION The prepared mucoadhesive buccal tablets of Atorvastatin calcium can help bypass extensive hepatic first-pass metabolism and improve bioavailability. The buccal bilayer tablets showed a mucoadhesion time of more than 6 h. Similarly, in-vitro permeation studies showed 85.68% drug release of the sustained dosage form. It can be concluded that formulation F2 could be used to release the drug unidirectionally in buccal cavity without the risk of mucosal irritation. ACKNOWLEDGEMENTS The authors wish to thanks UNICHEM Labs, Mumbai for providing Atorvastatin calcium as a gift sample and also to the Principal, Acharya & B. M. Reddy College of Pharmacy, Bangalore for providing necessary facilities to carry out the research work.

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