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I l of Pharmagenesis
2(1), January-June 2011, pp. 87-97
Formulation Development & Evaluation of Matrix Moderated
Transdermal System of Montelukast Sodium
Upendra Nagaich*1 and Vandana Chaudhary2
1Dept. of Pharmaceutics, B.V.M. College of Pharmacy, Shivpuri Link Road, Gwalior, Madhya Pradesh (India) 2Dept. of Pharmaceutics, Guru Gobind Singh College of Pharmacy, Yamuna Nagar, Haryana (India) Abstract: The main objective of the present study was to develop the matrix type transdermal films of Montelukast
Sodium and to evaluate them with respect to various in-vitro and in-vivo parameters. The matrix type transdermal systems were prepared by homogeneous dispersion of drug by using different ratios of HPMC K15M & EC polymers in 8:1, 4:1 and 2:1 ratios were increased to the total weight of 540 mg. HPMC K15M and EC were dissolved in 15 ml ethanol at room temperature, HPMC K15M as the rate-controlling membrane for all the systems.
The swelling properties of the polymers were studied, and drug-polymer interaction studies were performed.
The patches were subjected to various physicochemical studies, in-vitro release studies, permeation studies, and skin irritation studies. The optimized batch of HPMC K15M & EC 8:1 was selected to further study like effect of different natural permeation enhancers i.e. camphor, menthol and 1,8 cineole. This was done on the basis of in- vitro drug release studies. It was found that the batch F1, ratio 8:1 gave the best release rates which were found to increase with the increase in the concentration of the hydrophilic polymer HPMC K15M. All the patches with HPMC K15M: EC (8:1) showed controlled and sustained release. Formulation F5 containing 10% camphor was found to release the highest quantity of drug.
Keywords: Membrane controlled, hydrophilic polymer, swelling ratio, hydrophilic drug, in-vivo study.
Asthma (GINA) recommend use of anti- Asthma is the most common chronic disease and inflammatory controller therapy to attenuate its prevalence has substantially increased long-term inflammation and smooth muscle worldwide. It is associated with significant dysfunction in persistent asthma, thus preventing morbidity and economic burden. Chronic permanent airway changes. In this regard, inhaled inflammation and smooth muscle dysfunction are glucocorticosteroids (ICS) are the recommended consistent features of asthma pathophysiology. first line controller agents because of their broad The acute and chronic inflammation in asthma is anti-inflammatory properties. However, for some the result of extensive infiltration of the airway patients with persistent asthma, ICS fail to achieve by inflammatory cells including T cells, adequate response without increasing the dose eosinophils, mast cells and release of (2). These issues led to trials of adding alternative inflammatory mediators-cytokines and agents like long acting β-2 agonists (LABA), leukotrienes from these cells (1).
cromolyn sodium and leukotriene modifiers to achieve optimal disease control research into Current clinical practice guidelines from asthma pathophysiology enabled to understand National Asthma Education and Prevention the role of leukotriene as one of the pro- Programme (NAEPP) and Global Initiative for inflammatory agents in asthma. This initiated research and development of leukotriene Corresponding Author: Upendra Nagaich
modifiers as first mediator specific therapy for International Journal of Pharmagenesis, 2(1) 2011 Bronchial asthma is associated with coughing, dispersion using different ratios of HPMC K15M shortness of breath, chest tightness and wheezing. & EC polymers in 8:1, 4:1 and 2:1 ratios were There is contraction of airway smooth muscle, increased to the total weight of 540 mg. HPMC mucosal thickening from edema, cellular K15M and EC were dissolved in 15 ml ethanol infiltration and narrowing of the airway lumen due at room temperature, 50 mg of the drug was to thick viscid plugs of mucus. The bronchospasm dissolved in solution of HPMC K15M, and EC in asthma results from the release of various to obtain uniform solution, Dibutyl Pthalate 5% chemical mediators such as histamine, SRS-A, w/w of dry polymer was used as a plasticizers.
leukotrienes etc. The release of these agents might Backing membrane was cast by pouring and then occur as a result of antigen-antibody reaction or evaporating 4% aqueous solution of polyvinyl due to other pathological conditions (4).
alcohol in glass moulds covered on one side with The concept of delivering drugs through the aluminum foil, at 60 °C for 3 h. The uniform skin for systemic treatment of disease is gaining dispersion (3 ml each) was cast on the PVA increasingly great important due to its numerous backing membrane. Now, the polymeric solution advantages like bypassing hepatic first pass of the drug was poured on to the glass mould metabolism, enhancement of therapeutic (2.9 cm internal diameter) and dried at room efficiency, prolonged duration of action of drugs temperature in dust free environment. A release with short half life and maintenance of steady liner (wax paper) on either side of the film was plasma level of drug (5).
applied to complete the TDDS. The films were Treatment of chronic disease like asthma by stored in air tight container at ambient transdermal route of drug absorption might prove conditions for seven days prior to use, then out to have several advantages over other routes. of the three polymers combination the best was Therefore, the objective of the present work is to chosen for the further study by using three formulate a transdermal film for an anti-asthmatic different natural penetration enhancers Menthol, drug. Montelukast, which is a cysteinyl-leucotriene 1, 8 Cineole & Camphor. These enhancers were receptor antagonist, since it has half-life of 2-4 h, used as 5% & 10% (safe concentration) oral bioavailability, is around 64% (6).
concentration of total weight of the dry polymer.
The dry patches were kept in desiccators until Therefore, it is one of the most feasible drug use (7, 8).
delivery system for such a drug is transdermal film for the better management of the disease to Physical Evaluation
improve patient compliance. Recent years have witnessed a favorable preference among health Folding Endurance care givers for Montelukast, a leukotriene Folding endurance was determined by repeatedly antagonist in asthma management of children.
folding the film at the same place until it broke.
Materials and Method
The number of times the film could be folded at the same place without breaking was the folding Cadila endurance value (9).
Pharmaceuticals, Ahmedabad, HPMC K15M Zydus Cadila Ahmedabad, India, Ethyl Cellulose Weight Variation from Central Drug House (P) Ltd, Camphor from Karnataka fine chem. Menthol from Mora Weight variation was studied by individually Industrial Estate, Moradabad, 1,8 Cineole Redox weighing 10 randomly selected films. Such Pharmachem. Pvt Ltd Haridwar, India. Dibutyl determination was performed for each Pthalate from Karnataka fine chem. Ethanol: formulation (9).
DCM- Karnataka fine chem.
Film Thickness Preparation of TDDS Patches
The thickness of films was measured at three The matrix type transdermal films of Montelukast different places using a Dial caliper and mean Sodium were prepared by homogeneous values were calculated (9).
Formulation Development & Evaluation of Matrix Moderated Transdermal System of Montelukast.
Determination of Drug Content in the Film portion of a Wistar rat after killing the animal.
The uniformity of drug distribution was evaluated The hair was removed from the abdominal by determining drug content of the film by a portion with the help of scissor. The stratum spectrophotometric method. A known weight of corneum side of the skin was kept in intimate film was dissolved and diluted subsequently with contact with the release surface of the film under chloroform and the concentration of Montelukast test placed between the two halves of the sodium was spectrophotometrically measured at diffusion cell. The receiver phase was filled with 344 nm against the blank chloroform solution simulated phosphate buffer of pH 7.4 stirred at containing the same amount of polymer and 50 RPM on a magnetic stirrer. The whole plasticizer without drug (10).
assembly was maintained at 37 ± 0.5 °C. The skin was first stabilized until no ultraviolet (UV) Percentage of Moisture Content absorbance was observed (to eliminate the The films were weighed individually and kept in possibility of any interference on the permeation desiccators containing activated silica at room of drug through the skin by ammonia pre- temperature for 24 h. Individual films were treatment and due to other skin contents). The weighed repeatedly until they showed a constant amount of drug permeated was determined by weight. The percentage of moisture content was removing 5 ml samples at appropriate time calculated as the difference between initial and intervals up to 24 h. The volume was replenished final weight with respect to final weight (10).
with an equal quantity of pre-warmed receiver solution. The absorbance was read at 344 nm Percentage of Moisture Uptake spectrophotometrically. Cumulative amounts of drug diffused in mg/cm2 were calculated and A weighed film kept in desiccators at room plotted against time (11).
temperature for 24 h was taken out and exposed to 84% relative humidity (a saturated solution of Skin Irritation Studies aluminum chloride) in desiccators until a constant weight for the film was obtained. The percentage The hair on the dorsal side of Wistar rats was of moisture uptake was calculated as the removed by clipping 1 day before this portion of difference between final and initial weight with the experiment. The rats were divided into 4 respect to initial weight (10).
groups (n = 3). Group I served as the control, group II served as optimized formulation OF1, In-Vitro Drug Release Studies group III served as optimized formulation OF2 In-vitro drug release studies will be carried out and group IV received a 0.8% v/v aqueous using U.S.P. dissolution test apparatus II (Paddle solution of formalin as a standard irritant. A new over Disc method) into simulated phosphate buffer film or new formalin solution was applied daily of pH 6.8. The transdermal film was mounted on for 7 days. Finally, the application sites were the disc and placed at the bottom of the dissolution graded according to a visual scoring scale, always vessel. The dissolution medium was simulated by the same investigator (12).
phosphate buffer of pH 6.8 and the apparatus was maintained to 37 ± 0.5 °C. The apparatus was Temperature Dependent Stability Studies operated at 50 rpm and samples were withdrawn The purpose of stability testing is to provide at appropriate time intervals up to 24 h and evidence on how the quality of a drug substance analyzed at 344 nm spectrophotometrically. or drug product varies with time under the Cumulative % drug released were calculated out influence of a variety of environmental factors and plotted against time (11).
such as temperature, humidity and light and to establish a re-test period for the drug substance Ex-vivo Permeation Studies or a shelf life for the drug product and Ex-vivo permeation study was carried out using recommended storage conditions. To assess the Franz diffusion cell with a diffusion area of 3.14 drug and formulation stability, stability studies cm2. The skin was removed from the abdominal were done according to ICH guidelines Q1C.
International Journal of Pharmagenesis, 2(1) 2011 Stability studies were carried out on the films different ratios of HPMC K15M to EC (8:1, 4:1 of most satisfactory as per ICH Guidelines Q1C. and 2:1) from batch F1 to F9 as shown in Table The most satisfactory formulation stored in sealed 1. EC was added to improve the drug release by in aluminum foil. These were stored at room polymer swelling, elasticity and film forming temperature for 2 months. Films were evaluated properties of the patches. Butyl phthalate (5%) for in-vitro drug release, ex-vivo permeation study was added as plasticizer. PVA was used for the and various physical characteristics (13, 14).
casting of backing membrane. On the basis of physical characteristics and stability studies most suitable batch was selected for further Statistical significance of all the data generated studies of penetration enhancers like, Menthol, was tested by employing ANOVA followed by 1, 8 Cineole & Camphor (5% & 10% Newman-Keuls compare all pair of columns tests. concentration of total weight of the dry A confidence limit of P< 0.05 was fixed for polymer).
interpretation of the results using the software Prepared patches were smooth in appearance, PRISM (Graph Pad, Version2.01, San Diego, CA). uniform in thickness, mass, and drug content and Formulation F5 showed significant result when showed no visible cracks. More uniform thickness compared with all the other formulation (P<0.01). was shown by formulation F1 & F7(0.42 mm). The maximum folding endurance was shown by F1 formulation HPMC K15M: EC 2:1 which shows In the present study, Transdermal patches for that in above combination EC is responsible for controlled delivery of Montelukast Sodium were the strength.
developed using HPMC K15M as the base matrix.
F5 and F6 shows pH resembles that was more The folding endurance of the prepared patches resembles with skin pH. From physical evaluation was ranged from 120-125. The average weight F1 formulation compared with all different varies from 63.9±0.7 (F1) to 88.1± 0.1 (F9). F5 & F6 ranges of penetration enhancers as shown in shows more uniformity of content of (98.10±0.3, Table 2.
97.10±0.4). The thickness of the prepared patches Moisture content studies indicated that the was ranged between 0.35-0.42 mm.
increase in the concentration of hydrophilic Moisture content of the developed polymer was directly proportional to the increase formulations F­1 to F9 was varied from 2.50.00 to in moisture content of the films. The moisture 4.09.00%. Moisture uptake of the developed content of the prepared formulations was low, formulations F­1 to F9 was varied from 2.20 to which could help the formulations remain stable and reduce brittleness during long-term storage as shown in Table 2.
The stability studies showed that the formulation stored at 5.0 ± 3 ºC, 40 ± 2 °C, 75±5% Moisture uptake studies indicated that the RH and room temperature showed percentage increase in the concentration of hydrophilic cumulative amount of drug per cm2 of patch polymer was directly proportional to the increase 421.06, 418.01 and 416.91 µg respectively in 24 h in moisture uptake of the films. The moisture after 30 days, 419.80, 412.08 and 409.49.91µg uptake of the formulations was also low, which respectively in 24 h after 60 days, 407.31, 406.89 could protect the formulations from microbial and 404.89 µg respectively in 24 h after 90 days contamination and reduce bulkiness as shown in as shown in Fig. 5, 6, 7.
The optimized batch of HPMC K15M & EC 8:1 was selected to study the effect of different In the present study, Transdermal patches for natural permeation enhancers i.e. camphor, controlled delivery of Montelukast Sodium were menthol and 1,8 cineole. This was based on in- developed using HPMC K15M as the base vitro drug release study. It was found that the matrix. The patches were prepared using batch F9, ratio 8:1 gave the best release rate, which Formulation Development & Evaluation of Matrix Moderated Transdermal System of Montelukast.
was found to increase with the increasing The formulation F4 with 5% Camphor was concentration of the hydrophilic polymer HPMC found; show the best percentage cumulative drug K15M as shown in Fig. 1.
release of 422.01 mcg/cm2 in 24 h, due to its higher The in-vitro skin permeation of Montelukast fluidizing activity among the terpenes containing sodium from the selected TDDS through depilated essential oils.
goat abdominal skin was conducted using a Menthol produces local vasodilatation and modified Keshery-Chein diffusion cell. Percent could distribute preferentially into the cumulative amounts of drug permeated cm2 of intercellular spaces of stratum corneum and the patches were analyzed spectrophotometrically at possible reversible disruption of the intercellular 344 nm as shown in Table 3.
lipid domain (16) and 1,8 cineole improved the All the patches with HPMC K15M: EC (8:1) skin permeation of hydrophilic drugs better than showed controlled and sustained release. other terpenes and Montelukast sodium is Formulation F4 containing 5% camphor was hydrophobic in nature (17). Menthol was the most found to release the highest quantity of drug, effective permeation enhancer than the 1, 8 422.01 µg/cm2 of drug in 24 h as shown in Fig. 2. cineole.
All the formulations with and without Transdermal flux for different Montelukast permeation enhancers, F1 (without permeation sodium formulations with natural permeation enhancer), F4 and F5 (camphor- 5%, 10%) as enhancers depilated mouse abdominal skin was shown in Fig. 2, F6 and F7 (menthol 5%, 10%) as conducted using a modified Keshary-chein shown Fig. 3, F8 and F9 (1, 8 cineole 5%, 10%) as diffusion cell Table 3. The flux from drug-polymer shown Fig. 4, appeared to follow similar patterns TDDS (without permeation enhancer) was less of drug release profiles, i.e. initially apparent zero- release of drug than that obtained after order and then first order release kinetics. Initially application of camphor, menthol, 1,8 cineole. The for first few hours the drug release, kinetic scanning electron microscopy (SEM) is carried out patterns followed zero-order drug release profiles for formulation F5 as shown in Fig. 8, 9, 10, 11.
and with the enhancement of time the release The data indicated that the transdermal flux profiles gradually changed into the concentration of Montelukast sodium increased with presence dependent first order release kinetics as shown of 5% of camphor than comparison of other penetration enhancer in the formulation. Hence The effect of permeation enhancers often Camphor is better permeation enhancer than depends on their applied concentrations (11). menthol and 1,8 cineole as shown in Fig. 12.
The mechanism of action of permeation Formulation F4 subjected to stability studies enhancers (7) are-: (i) disruption of the highly was stored at 5 ± 3 ºC, room temperature and 40 ordered structure of SC lipids, (ii) interactions ± 2ºC, 75% RH for three months. Optimal storage with intracellular proteins or (iii) improvement conditions of the formulation were assayed by in partitioning of the drug, co-enhancers or co- analyzing the percentage cumulative amount of solvent into the stratum corneum. From the drug permeated per cm2 of patch after time studies it was reported that terpenes enhance interval of 30, 60, 90 days. It was determined diffusion of drugs by extracting lipids from by plotting graph between percentage stratum corneum (14, 15) which results in cumulative amount of drug per cm2 of patch and reorganization of lipid domain and barrier time intervals that reflected kinetics of release disruption (16, 17). The mechanism of barrier profile.
disruption may be due to the competitive hydrogen bonding of oxygen containing The percentage cumulative amount of drug monoterpenes with ceramide head groups, from the formulation stored at 2–8ºC was found thereby breaking the interlamellar hydrogen- higher in comparison with the formulation stored bonding network of lipid bilayer of stratum at room temperature, and 40 ± 2ºC, This indicated corneum and new polar pathways or channels that the formulation tend to degrade faster at are formed (17).
higher temperature.
International Journal of Pharmagenesis, 2(1) 2011 Composition of Prepared Films
Different Combination of HPMC K15m: Ec ETHANOL: DCM (1:1) Surface pH *All values are expressed as mean ± SD (n = 6) In- vitro Skin Permeation Profile of HPMC K15M: EC Patches
Cumulative Drug Release(mcg/sq.cm2) *Significant compared with all other formulation (P < 0.01).
Formulation Development & Evaluation of Matrix Moderated Transdermal System of Montelukast.
Permeation Kinetic Studies of Various Batches
Kinetic Profile For Higuchi For Pappas For 1st order Equation For Zero Order of formulations Stability Studies Physical Characteristic of Best Formulation F4 during Stability Studies
endurance Room temperature 20±2 Room temperature 21±3 Room temperature 20±3 Figure 1: Plot of Cumulative Drug Release Vs Time Profile
International Journal of Pharmagenesis, 2(1) 2011 Figure 2: Skin Permeation Studies of HPMC K15M & EC (F-9, 8:1) Combinations with 5% Camphor as Permeation Enhancer
Figure 3: Skin Permeation Studies of HPMC K15M & EC Combinations with Menthol as Permeation Enhancer
Figure 4: Skin Permeation Studies of HPMC K15M & EC Combinations with 1,8 Cineole as Permeation Enhancer
Formulation Development & Evaluation of Matrix Moderated Transdermal System of Montelukast.
Figure 5: % Cumulative Amount Release Profile of Formulation F5 during Stability Studies
Figure 6: % Cumulative Amount Release Profile of Formulation F5 during Stability Studies
Figure 7: % Cumulative Amount Release Profile of Formulation F5 during Stability Studies

International Journal of Pharmagenesis, 2(1) 2011 Figure 8: Scanning Electron Microscopy of Normal F5 Film Figure 11:Scanning Electron Microscopy of F5 Film after 8
at X300 Magnification h skin Permeation at X1000 Magnification Figure 9: Scanning Electron Microscopy of Normal F5 Film
at X1000 Magnification Figure 12:In-vitro Skin Permeation Studies
In the present study, an attempt was made to
develop unidirectional transdermal films of Montelukast sodium for treatment/management of asthma. Montelukast sodium transdermal delivery system prepared by matrix diffusion Figure 10:Scanning Electron Microscopy of F5 Film after 8
type technique with different range of compatible h skin Permeation at X300 Magnifications polymers like HPMC K15M as a rate controlling Formulation Development & Evaluation of Matrix Moderated Transdermal System of Montelukast.
polymer and EC as a membrane modulated [8] Martin, A., Bustamante, P. & Chun, A. H. C. (1999), polymer. For the more skin penetration of the Physical Pharmacy, 4th ed. B.I. Waverly Pvt. Ltd, New drug to the body natural permeation enhancers Delhi: 284-317.
was also added. So, with the addition of [9] Aqil, M., Asgar, A. (2002), Monolithic Matrix Type permeation enhancer it can be proved as good Transdermal Drug Delivery Systems of Pinacidil Monohydrate: in-vitro Characterization, European delivery system for the effective treatment of Journal of Pharmaceutics and Biopharmaceutics, 54(2): 161-
asthma with the aid of patient compliances.
[10] Alagusundaram M., Chetty C. M., Dhachinamoorthi D.
Development and Evaluation of Novel-trans- buccoadhesive Films of Famotidine. J. Adv. Pharm. Tech. The authors are thankful to Cadila Pharmaceuticals, Res. 2011; 2: 17-23.
Ahmedabad (Gujarat) for providing gift sample of drug. The [11] Dandagi, P. M., Manvi, F. V. (2003), Formulation of Chairman and management committee members of B.V.M.
Transdermal Drug Delivery System of Ketotifen College of Pharmacy, Gwalior (M.P.) are gratefully Fumarate, Indian Journal of Pharmaceutical Science, 65(3):
acknowledged for providing necessary facilities to carry out [12] Nagaich, U., Chaudhary, V., Sharma, P. & Yadav, A.
(2009), Formulation and Development of Metoprolol Tartrate Bucco-adhesive Films, The Pharma Research, [1] Walia, M., Lodha, R. & Kabra S. K. (2006), Montelukast 1(1), 41-53.
in Pediatric Asthma Management, Indian Journal of [13] Amnuaikit, C., Ikeuchi, I., Ogawara, K., Higaki, K. & Pediatrics, 73(4): 275-82.
Kimura, T. (2005), Skin Permeation of Propranolol from [2] Tripathi, K. D. (2003), Drugs for Bronchial Asthma. In: Polymeric Film Containing Terpene Enhancers for Essential of Medical Pharmacology. 5th ed.; 205.
Transdermal use, International Journal of Pharmaceutics, [3] Aulton, M. E. (2002), Pharmaceutics: The Science of Dosage Form Design. 2nd ed. Published by Livingstone [14] Mishra, A. N. (1997) Controlled Novel Drug Delivery C. Elsevier science Ltd. 2002:315-320.
Ed. Jain NK, CBS Publication, New Delhi. 1st ed.: 100- [4] Robinson, J. R., Lee, V. H. L. (1987), Controlled Drug Delivery: Fundamentals and. Applications, 2nd ed. [15] Mehdizadeh, A., Toliate, T., Rouini, R. M., Abashzadeh, Published by Marcel Dekker, Inc. New York. 29: 253- S. & Dorkoosh, F. (2004), Design and in vitro Evaluation of New Drug-in-adhesive Formulations of Fentanyl [5] Jain, N. K. (1997), Transdermal Drug Delivery. In: transdermal Patches, Acta Pharmaceutica, 54: 301-317.
Controlled and Novel Drug Delivery. 1st ed.: 100-114.
[16] Bagyalakshmi, J., Purpa R. & Rajappam V. (2007), [6] Chein, Y. W. (2006), Transdermal Drug Delivery. In: Formulation Development and in-vitro and in-vivo Novel Drug Delivery System. 2nd ed.; 50: 338-344.
Evaluation of Membrane-moderated Transdermal Systems of Ampicillin Sodium in Ethanol: pH 4.7 buffer [7] Arora, P., Mukherjee, B. (2002), Design, Development, Solvent System, AAPS PharmSciTech, 8(1), E50-E55.
Physicochemical, and in-vitro and in-vivo Evaluation of Transdermal Patches Containing Diclofenac [17] Chien, Y. W. (1987), Development of transdermal drug Diethylammonium Salt, Journal of Pharmaceutical delivery system, Drug Development Industrial Pharmacy, Science, 91(9): 2076-89.
13: 589-651.



The International Journal of Biochemistry & Cell Biology 38 (2006) 1975–1985 Co-expression of chaperonin GroEL/GroES enhances in vivo folding of yeast mitochondrial aconitase and alters the growth characteristics of Escherichia coli Parul Gupta, Nishtha Aggarwal, Pragya Batra, Saroj Mishra, Tapan K. Chaudhuri Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology, Delhi,

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