Compatibility study In-vitro drug release Study of Solid Lipid Nanoparticle Based Transdermal Drug Delivery System for Rasagiline Mesylate

 

Prashant S. Wake¹*, Dr. M. D. Kshirsagar²

¹Research Scholar, P. Wadhwani College of Pharmacy, Yavatmal (M. S.)

²Professor, P. Wadhwani College of Pharmacy, Yavatmal (M. S.)

 

ABSTRACT:

Transdermal drug delivery system (TDDS) is the dosage forms which deliver a therapeutically effective amount of drug across a patient’s skin. The Solid lipid nanoparticles were successfully developed for rasagiline mesylate. SLN dispersions were prepared by melt emulsification and solidification at low temperature method. Compatibility between Drug and polymer study by FTIR. DSC and in-vitro release profile were carried out. In the Transdermal Drug Delivery System, Formulations F1-F9 was prepared by solvent casting method using 1.5%, 2.5% and 3.5% of HPMC K4M and 20%, 30% and 40% (w/w of dry polymer) of PEG 400. The formulation F5 was selected as the promising formulation on the basis of cumulative % drug release. The cumulative % drug diffused of F5 was found to be 89.55 ± 1.983. Further, the patch was found to be free of skin irritation. From the results stability study it can be concluded that the patches can be stored at 40°C and 75% RH without any significant stability problems. The formulation satisfied all the pharmaceutical parameters of transdermal films and appears to be promising.

 

 

 

1. INTRODUCTION:

Transdermal drug delivery system (TDDS) is the dosage forms which deliver a therapeutically effective amount of drug across a patient’s skin. It increases patient compliance and avoid first pass metabolism over injectables and oral routes. Solid lipid nanoparticles are at the forefront of the rapidly developing field of nanotechnology with several potential applications in drug delivery, clinical medicine and research as well as in other varied sciences.

 

SLN as colloidal drug carrier combines the advantage of polymeric nanoparticles, fat emulsions and liposome; due to various advantages, including feasibility of incorporation of lipophilic and hydrophilic drugs, improved physical stability, low cost, ease of scale-up, and manufacturing. SLNs are prepared by various advanced techniques. The site specific and sustained release effect of drug can better achieved by using SLNs. Nanoparticles have been used extensively for applications in drug discovery, drug delivery, and diagnostics and for many others in medical field.^1-2

 

Present Study was done to prepare the optimize formulation of SLN Loaded Transdermal Drug Delivery System containing Rasagiline Methysylate. Rasagiline is used to treat symptoms of Parkinson's disease both alone and in combination with other drugs. It has shown efficacy in both early and advanced Parkinsons, and appears to be especially useful in dealing with non-motor symptoms like fatigue³.

2. MATERIAL AND METHOD:

Rasagiline Mesylate was supplied as gift sample by Anazeal Research Pvt Ltd, Mumbai, India. Stearic acid, tween 80, Methanol, Acetone, HPMC K4M, Propylene Glycol 400, DMSO and IPA were procured from from S.D. fine-chem Ltd. All other ingredients were of analytical reagent grade.

 

Fig 1 FTIR spectra of Rasagiline Mesylate

 

Fig 2 FT-IR spectrum of FITR spectra of (A) pure Rasagiline mesylate (B) stearic acid (C) mixtures of Rasagiline mesylate with stearic acid

 

3. Experimental Details:

3.1 Compatibility study between drug and polymer

(A) FTIR Spectrophotometry Analysis

Data acquired from FTIR Spectrophotometric studies of drug-excipients mixtures stored at 25°C±2/60%±5RH, indicates no significant changes in the spectra. The pure Rasagiline mesylate characteristic spectra were shown, a band of 2934 cm^-1and 2849 cm^-1owing to Aromatic C-H stretching and Aliphatic C-H stretching groups. The other bands Peaks at 3219 cm^-1 (Secondary amine N-H) and 1192 cm^-1(Aliphatic amino C-N) respectively. The pure Rasagiline mesylate spectrum is shown in Fig. 1. Fig. 2 shows the FTIR spectra of Rasagiline mesylate, Stearic acid and mixtures of Rasagiline mesylate and Stearic acid. The characteristic band Peaks at around 2849 cm^-1 and 1702 cm^-1 in Stearic acid are assigned to the Aromatic C-H stretching and C=O Asymmetric stretching, respectively. The peaks present at 2674 cm^-1 and 1463 cm^-1 owing to Carboxylic O-H and CH₂ bending. Mixtures of Rasagiline mesylate and Stearic acid spectra showed no change in the positions of the bands at 1701 cm^-1 (Imide), 2917 cm^-1(Aromatic C-H stretching), 2805 cm^-1 (Aliphatic C-H stretching) and 1191 cm^-1 (Aliphatic amine C-N) in Rasagiline mesylate. The FTIR spectra of combination of drug with the polymer did not show any changes in the characteristic peaks of the Rasagiline Mesylate. The specific peaks at wave number1479.45 cm-1 due to - CH2- bending, at 646.17 cm-1 due to S- O bending, at 3279.10 cm-1due to ≡ C-H stretching, at 2125.63 cm-1 due to C ≡ C stretching, 1626.05, 1604.83, 1560.46 cm-1due to N-H bending (secondary amines) remain unchanged indicating that the drug had not interacted with the polymer.

 

(B) Differential scanning calorimetric analysis (DSC)

The DSC thermogram of Rasagiline mesylate showed an endothermic peak at 131.49°C (It represents the dehydration of bound water) and 155.56°C (melting point). Rasagiline mesylate-Stearic acid mixtures showed the endothermic peak at 63.63°C and 153.97°C (Fig.4). The melting endothermic peak of Rasagiline mesylate at 155.56°C in Rasagiline mesylate-Stearic acid mixture confirms there is no interaction between Stearic acid and Rasagiline mesylate.

 

Fig 3: DSC thermogram of Rasagiline mesylate

 

Fig 4 DSC thermogram of Rasagiline mesylate  with stearic acid

 

3.2 Methodology of the Experiment⁴:

SLNs loaded with Rasagiline Mesylate were prepared using melt emulsification and low-temperature Solidification method. Rasagiline Mesylate was dissolved in methanol and mixed with acetone solution containing stearic acid. The mixtures were sonicated for 15 minute, and then added drop wise to Tween 80 solution, stirred at 3000 rpm for 0.5 h at 70°C temperature. The mixed solution was transferred to icy water bath and stirring for four hour at 3000 rpm. Different formulations of drug loaded SLN were prepared by varying concentrations of stearic acid as shown in the below Table 1.

 

Table 1 Composition of solid lipid nanoparticles formulation containing Rasagiline

Ingredients	F1	F2	F3	F4	F5	F6
Rasagilline Mesylate (mg)	100	100	100	100	100	100
Stearic Acid (mg)	1000	1250	1500	1000	1250	1500
Tween 80 (%)	2.5	2.5	2.5	2.0	2.0	2.0
Methanol (ml)	1	1	1	1	1	1
Acetone (ml)	1	1	1	1	1	1

 

3.3 Formulation of transdermal patch⁵

Transdermal patches were prepared by solvent casting method. In this method polymeric solution of HPMC K4M was prepared by dissolving weighed quantity of HPMC K4M in IPA. Then calculated quantity of PEG 400 was added and mixed well till clear solution was obtained. The resultant solution was allowed to stand till all entrapped air bubbles get removed. Then solution was poured into a clean and dry glass petri dish containing DMSO and allowed to dry. The dried patches were carefully removed from the petri dish, checked for any imperfections or air bubbles and cut in to pieces of 4 cm².

Table 2 Composition of Transdermal Patches of formulation containing SLN of Rasagiline

 

4. Evaluation

4.1 Evaluation of solid lipid nanoparticles^6-7

4.1.1 In vitro drug release

The in vitro drug release study of solid lipid nanoparticles was performed by locally fabricated Franz diffusion type cell. The study was performed at 30±2°C temperature. Receptor compartment of diffusion cell contained 30 ml 30% v/v PEG 400 in phosphate buffer (pH-6) solution and was constantly stirred by a magnetic stirrer (Expo India Ltd., Mumbai, India) at 50 r/m. Dialysis membrane (molecular weight cut off 12 KD, HiMedia Laboratories Pvt. Ltd., India) was employed as release barrier in between receptor and donor compartment which was previously was with distilled water and soaked with 30% v/v PEG 400 solution. Time to time 5 ml samples was withdrawn through the sampling port of the diffusion cell in intervals one h, over 8 h. same amount of 30% v/v PEG 400 solution was replaced immediately. The collected samples were suitably diluted and analyzed by HPLC (Shimadzu, Japan).

 

4.2 Evaluation of Transdermal Drug Delivery Systems^8-10

4.2.1 In vitro drug release studies

The in-vitro permeation study of fabricated transdermal patches of ketoprofen was carried out by using excised rat abdominal skin and franz diffusion cell. The skin was sandwiched between donor and receptor compartments of the diffusion cell. A 2.2 cm diameter patch was placed in intimate contact with the stratum corneum side of the

 

skin; the top side was covered with aluminum foil as a backing membrane. Teflon bead was placed in the receptor compartment filled with 12ml of normal saline. The cell contents were stirred with a magnetic stirrer and a temperature of 37 ± 5°0C was maintained throughout the experiment. Samples of 1ml were withdrawn through the sampling port at different time intervals for a period of 24 h, simultaneously replacing equal volume by phosphate buffer pH 7.4 after each withdrawal. The samples were analyzed spectrophotometrically at 268.81 nm.

 

4.2.2 Stability studies as per ICH guidelines.

Stability studies will be conducted according to the ICH guidelines by storing the TDDS samples at 40±0.5°c and 75±5% RH for 6 months. The samples will be withdrawn at 0,30,60,90 and180 days and drug content will be analyzed.

 

5 RESULTS AND DISCUSSION

5.2.1 In vitro drug release of Solid Lipid Nanoparticle

Table 3:- in vitro % drug release of formulations

Time (hours)	F1	F2	F3	F4	F5	F6
0	0	0	0	0	0	0
2	9.34	11.33	13.55	11.71	8.52	7.35
4	12.51	13.49	16.85	14.56	11.47	9.80
6	15.22	18.26	20.52	18.44	14.65	12.25
8	18.18	21.56	24.98	22.63	17.48	15.65
10	21.25	24.50	29.22	26.66	20.26	17.46
12	23.58	28.85	32.55	30.33	23.55	21.2
16	29.74	32.85	40.22	35.42	29.41	26.65
24	36.55	38.25	49.83	42.58	34.55	31.11

Figure 5 In vitro release study SLN formulations

 

5.3 In vitro drug diffusion of transdermal drug delivery systems

Table 4 In vitro drug release data of factorial formulations F1 to F4

*Mean value ±SD (n=3)

 

Table 5 In vitro drug release data of factorial formulations F5 to F9

Time (hr)	F5*	F6*	F7*	F8*	F9*
0	0.00 ± 0.000	0.00 ± 0.000	0.00 ± 0.000	0.00 ± 0.000	0.00 ± 0.000
3	18.38 ± 1.956	21.66± 3.198	18.08± 2.816	17.68 ± 1.684	16.67 ± 3.428
5	38.49 ± 2.491	48.85± 1.365	31.68± 1.648	33.64 ± 2.880	33.65 ± 2.038
7	55.49 ± 1.955	56.98± 2.384	45.55 ± 3.284	43.62± 1.658	55.35 ± 2.490
9	67.37 ± 0.983	71.62± 0.934	61.38 ± 0.584	62.95± 2.067	67.48± 0.398
12	82.54 ± 2.097	80.66± 2.008	70.50 ± 2.907	76.64 ± 0.981	74.39± 1.387
24	89.55± 1.983	83.66± 1.258	81.95 ± 1.648	84.65± 2.437	85.68 ± 1.982

*Mean value ±SD (n=3)

 

Figure 6: In vitro diffusion study of F1 to F9 batch

5.5 Stability Study

The promising formulation F5 was subjected at 40 ± 0.5°C temperature and 75 ± 5 % RH for 1 month to check the stability. The results of thickness, drug content, folding endurance and other parameters after 1 month storage of prepared transdermal patches are shown in table 6.

 

Table 6 %Cumulative drug release study of F5 at 0 day and after 30 days %Cumulative drug release

*Mean value ±SD (n=3)

 

Figure 7 In vitro diffusion study of F5 at 0 day and after 30 days

 

5. CONCLUSION:

The Solid lipid nanoparticles were successfully developed for rasagiline mesylate. SLN dispersions were prepared by melt emulsification and solidification at low temperature method. Compatibility between Drug and polymer study by FTIR, DSC and in-vitro release profile were carried out. It was seen that increasing the stearic acid concentration led to higher entrapment and by increasing the concentration of tween 80 lead to smaller the particle size. In-vitro drug release pattern of SLN showed fast and control release. Immediate releases as well as sustained release both are of interest for topical application. Immediate release can be useful to improve the penetration of drug and maintain the concentration

 

work as loading dose, while sustained release supplied the drug over a prolonged period of time.

 

In the Transdermal Drug Delivery System, Formulations F1-F9 was prepared using 1.5%, 2.5% and 3.5% of HPMC K4M and 20%, 30% and 40% (w/w of dry polymer) of PEG 400. The formulation F5 was selected as the promising formulation on the basis of cumulative % drug diffusion. The cumulative % drug diffused of F5 was found to be 89.55 ± 1.983. Further, the patch was found to be free of skin irritation. From the results stability study it can be concluded that the patches can be stored at 40°C and 75% RH without any significant stability problems. The formulation satisfied all the pharmaceutical parameters of transdermal films and appears to be promising, would be able to offer benefits such as sustained drug release, reducing frequency of administration, improving bioavailability, and thereby may help to improve patient compliance.

 

6. REFERENCES:

1.       Bharkatia M., Nema R. K., Designing and Evaluation of Transdermal Patches of Nimesulide, Asian J. Pharm. 1(1), 2006, 46-48.

2.       Gupta S. P., Jain S. K., Effective and controlled transdermal delivery of metoprolol tartarate, Indian J. Pharm. Sci., 67(3), 2005, 346-350

3.       Aggarwal G., Dhawan S., Psychotropic Drugs and Transdermal Delivery: An Overview, International Journal of Pharma and Bio Sciences. 2010: 1-12.

4.       American Psychiatric Association: Diagnostic and statistical Manual of Mental Disorders, Fourth Edition. Washington, DC: American Psychiatric Association, 1994.

5.       Bagdiya O., Purnima A., Formulation and Development of Venlafaxine Hydrochloride Extended Release Tablet and invitroCharacterizations. International Journal of PharmTech Research. 2012 (4) 4: 1777-1784.

6.       Bhandari R., Kaur  P., A Method to Prepare Solid Lipid Nanoparticles with Improved Entrapment  Efficiency of Hydrophilic Drugs. Current Nanoscience, 2013(9) 1573-4137.

7.       Das M.K., Bhattacharya A., Ghosal S.K., Transdermal delivery of Trazodone hydrochloride from acrylic films prepared from aqueous latex. Int J Pharm sci 2006; 68(1); 41-46

8.       Dewalkar H., Hariprasanna R.C., kulkarni U., Design and development of fast dissolving tablets containing ziprasidone by solid dispersion method JPSBR: 2012 :18-24

9.       Aggarwal G., Dhawan S., Development, Fabrication and Evaluation of Transdermal Drug Delivery System - A Review Pharm Res. 2009.

10.     Shahi S. R., Somani V. G., Shamkuvar P. B., Kale M. A., Kataria M. V., Effect of enhancers on topical delivery of ketorolac tromethamine. Asian J. Pharmaceutics, 1, 2007, 184-186.

 

 

 

 

Received on 26.03.2017       Accepted on 02.05.2017     

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