Formulation and Evaluation of Immediate Release Film Coated Tablets of An Anticancer Drug (Dasatinib)

Arti Mohan¹, G. Sangeetha²

¹Depatment of Pharmaceutics, Krupanidhi College of Pharmacy

²Department of Pharmaceutics, Gautham College of Pharmacy, Bengaluru, Karnataka, India.

*Corresponding Author E-mail: artimohan89@gmail.com

ABSTRACT:

Aim: The objective of the present study was to develop an immediate release tablet of Dasatinib using different concentration of superdisintegrants with a view to obtain rapid action for myelogenous leukemia and acute lymphoblastic leukemia^[1]. Dasatinib is a chemotherapy medication used to treat certain cases of leukaemia. Materials and Methods: Various formulations were prepared by direct compression technique using super disintegrants (cross carmellose sodium, sodium starch glyclote and crospovidone). Effect of different superdisintegrants on drug release was studied. Optimized formulation was selected on the basis of drug release and its in-vitro release profile compared with the reference product (Sprycel). Results and Discussion: The dissolution results showed gradient increase in the drug release with the increase in concentration of the super-disintegrants. Conclusion: Among all the formulations F3 containing croscarmellose sodium (4.5%) was found to show best results with 98.3% release within 45 minutes and it exhibited a release profile comparable to the reference product.

KEYWORDS: Dasatinib, Myelogenous leukemia, Croscarmellose sodium, Direct compression technique, Immediate release.

INTRODUCTION:

The oral route of drug administration is one of the most popular routes of drug administration. Immediate release tablets are designed to disintegrate and release their medicaments with no special rate controlling features such as special coatings and other techniques. Immediate release dosage forms are most commonly formulated when rapid response is required. This also serves as an advantage for patient compliance. Superdisintegrant are the vital components of Immidiate release dosage forms along with various common excipients like diluents, binder, lubricants, glidant etc.

Direct compression method for formulation ofmidiate release tablets is more economic, reducing the cycle time and straight forward in terms of good manufacturing practice requirements. So pharmaceutical industry is now focusing increasingly on this process.

Dasatinib, sold under the brand name Sprycel, is a chemotherapy medication used to treat certain cases of myelogenous leukemia and acute lymphoblastic leukemia^[1]Dasatinib has become first line drug in the pharmacotherapy of patients with chronic mylogenuous leukemia (CML). This is because the drug possesses tolerability and safety advantages over the other tyrosine kinase inhibitors. Dasatinib (DST) is a BCS Class II drug having very low solubility and high permeability. Animal data suggests that the absolute bioavailability of DST is about 14 to 34% due to an extensive first-pass effect.^[2] Immidiate release tablets can overcome the extensive first pass effect as they disintegrate and are absorbed through the oral cavity therby bypassing the extensive first pass effect.

The basic objective of present studies formulation and in vitro evaluation studies of immediate release tablets of DST using super disintegrant like cross carmellose sodium, sodium starch glyclote and crospovidone with a view to obtain rapid disintegration when taken through oral route, permitting a rapid onset of action for cancer therapy.

MATERIALS AND METHODS:

Materials:

Dasatinib was a gift sample from NATCO Pharma Ltd (Hyderabad, India) and Lactose monohydrate was a gift sample from Sisco research Labs, Pvt Ltd, Mumbai and Crosscarmellose sodium, Sodium starch glycolate and Cross Povidone were procured from SD Fine Chemicals ltd (Mumbai, India), Kamarlal and Co, Hyderabad and Anshul Agencies, Mumbai respectively. All other chemicals and reagents used were of analytical grade.

Methods:

Formulation of Immidiate Release tablets.^[3]:

Dasatinib monohydrate, crosscarmellose sodium /sodium starch glycolate /crospovidone, lactose monohydrate, hydroxypropyl cellulose and microcrystalline cellulose (PH 102) were sifted through sieve no 40 and blended for 2 min. To this magnesium stearate (previously passed through sieve no 60) was added and blended for 1 min and compressed using Tablet Compression Machine-8 station by using 12.5 mm round punches.

Preparation of coating solution:

Opadry white and water were weighed accurately. Opadry was dispersed in water under constant stirring and stirred for 45 mins. The solution was filtered and used for coating.

Film Coating:

10% w/v coating solution of opadry white was used.

Experimental study:

Solubility:

The solubility of the drug sample was carried out in different solvents (aqueous and organic).^[4]

Moisture Content:

50 ml of methanol was taken in titration vessel of Karl Fischer titrator and titrated with Karl Fischer reagent to end point. The DST pellets were ground to fine powder, 0.5 g of the sample was weighed and transferred quickly to the titration vessel, stirred to dissolve and titrated with Karl Fischer reagent to the end point.

It was estimated according to the formula:

V x F x 100

Moisture content =----------------------

Weight of the Sample in mg

Where,

V= volume in ml of Karl Fischer reagent consumed for sample titration.

F= factor of Karl Fischer reagent.

The samples were checked for description, related substance and water content by KF.

Preparation of the Standard curve:

10 mg of DST was accurately weighed and transferred to 100 ml of volumetric flask Acetate buffer (pH4) was added and volume made upto 100 ml. Aliquots of 0.2 to 2.0 ml were withdrawn and diluted upto 10 ml with distilled water (2 to 20 µg/ml). Absorbance was measured at 320 nm using UV spectrometer (Shimadzu 1800, Japan) and graph of absorbance v/s concentration was plotted^.).^[5]

Drug-Excipient Compatibility Studies:

DST and excipientswere mixed in different ratios in a polybag for 5 min. Each mixture was allotted sample code for identification and the mixtures were thoroughly sealed in glass vials and checked for description, related substance and water content by KF and evaluated initially, after 14 days at 55⁰C ± 2⁰C and after 28 days at 40 ± 2⁰C and 75 ± 5% RH for related substances by HPLC.^[6]

Evaluation of Pre-compression parameters:

Various pre compression parameters like Angle of Repose, Bulk Density, Tapped Density, Carr’s Index and Hausner’s Ratio for the formulation blends were estimated.^[7]

Evaluation of Post-compression parameters.^[8]

Weight variation test:

Twenty tablets were selected at random and average weight was determined. Then individual tablets were weighed and individual weight was compared with an average weight. Not more than one tablet should fall outside this range.

Hardness test:

10 tablets from each batch were selected and hardness was measured using Digital hardness tester to find the average tablet hardness or crushing strength. Hardness of 4 kg is considered suitable for handling the tablets.

Friability (%):

Twenty tablets were weighed accurately and placed in Friabilator for the given specification (4 min at 25 rpm). The tablets were weighed again and % friability was then calculated.

Formulation of Immidiate Release Film Coated tablets of Dasatinib:

Table 1: Various formulation trails of Dasatinib Immediate Release Tablets

S. No	Ingredients	F1	F2	F3	F4	F5	F6	F7	F8	F9
1	Dasatinib Monohydrate	70mg	70mg	70mg	70mg	70mg	70mg	70mg	70mg	70mg
2.	Microcrystalline cellulose (PH 102)	395.32	347	299.77	395.32	347	299.77	395.32	347	299.77
3.	Lactose monohydrate	43.92	86.75	128.48	43.92	86.75	128.48	43.92	86.75	128.48
4.	Croscarmellose sodium	13.75 (2.5%)	19.25 (3.5%)	24.75 (4.5%)	___	___	___	___	___	___
5.	Sodium starch glycolate	___	___	___	13.75 (2.5%)	19.25 (3.5%)	24.75 (4.5%)	___	___	___
6.	Crospovidone	___	___	___	___	___	___	13.75 (2.5%)	19.25 (3.5%)	24.75 (4.5%)
7.	Hydroxypropyl cellulose	22 mg	22 mg	22 mg	22 mg	22 mg	22 mg	22 mg	22 mg	22 mg
8.	Magnesium stearate	5 mg	5 mg	5 mg	5 mg	5 mg	5 mg	5 mg	5 mg	5 mg
	Total weight	550 mg	550 mg	550 mg	550mg	550 mg	550 mg	550 mg	550 mg	550 mg

Disintegration Test:

Disintegration test, measured using USP tablet disintegration test apparatus (ED2L, Electro lab, India) using 900 ml of distilled water at room temperature (37±2⁰C). Disintegration time was measured for six tablets by inserting each tablet in each disk

Uniformity of Drug Content:

Assay of Immidiate Release Film Coated tablets of DST was done to find out the amount of drug present in one tablet. For this test, 20 tablets were weighed and powdered in a glass mortar and powder equivalent to 100 mg of drug was placed in a stoppered 250 mL volumetric flask and dissolved in 160 mL of a mixture of methanol and acetonitrile in ratio of 50 : 50 v/v respectively and sonicated for 30 min with occasional shaking. The resulting solution was cooled, diluted to volume with the solvent mixture and centrifuged at 3000 rpm for 15 minutes. 5 ml of the above solution was transferred to 100 ml volumetric flasks and diluted to volume with the diluents. The concentration of drug was determined by High performance liquid chromatography equipped with UV detector and data handling system and analyzed at 320 nm to calculate the % of DST in DST tablets. Chromatographic separations were performed using a Cosmicsil BDS C18 (150 ₓ4.6 mm) 5 µm column maintained at 35̊ C. Injection Volume was 10 µL and run time was 12 mins. The flow rate was maintained at 1 ml/min. Chromatogram was recorded and the peak area response for major peaks was measured.

In vitro Dissolution studies:

In vitro dissolution test was carried out by using USP type II (paddle) apparatus. 1000 ml of acetate buffer pH 4 with 1 % triton X-100 was used as dissolution medium and the paddle was rotated at 60 rpm at temperature (37^oC ± 0.5^oC). Sampling was done at 10, 15, 30 and 45 min and was replaced by media after each sampling interval. The samples were then analysed spectrophotometrically at λmax of the drug.^[9]

Comparison of in vitro percentage drug release of Optimized formulation (F3) and innovator/ marketed product:

In vitro dissolution test was carried out to compare the optimized formulation (F3) with the marketed product (Sprycel).^[10]

Stability Studies:

The purpose of stability testing is to provide evidence of the quality of the drug substance or drug product and how it varies with time under the influence of a variety of environmental conditions (heat, humidity, light, air etc). The optimized formulation was packed in suitable packing and kept at different temperature, humidity conditions and the samples were analyzed for their physical and chemical properties.^[11]

RESULTS AND DISCUSSION:

Solubility:

Practically insoluble in aqueous media, slightly soluble in ethanol and soluble in methanol. The solubility of DST was found to be 3.4 mg/ml in ethanol, 18 µg/ml at pH 2.6, 8 µg/ml at pH 6 and < 1µg/ml at pH 7.4.

Moisture Content:

Water content in the sample was found to be 2.8%

Drug-Excipient Compatibility Studies:

No Characteristic change in the colour of the powder and no additional degradation of the product were observed as shown in Table 2

Preparation of the Standard curve:

Calibration curve of Dasatinib in acetate buffer was found to be linear and obeyed Beer’s law in the concentration range of 2 to 20 µg/ml as shown in Figure 1.

Evaluation of Pre-compression parameters:

All the pre compression specifications were found to be within the prescribed limits according to IP, thus ensuring good flow property of the formulation blends as shown in Table 3.

Table 2: Drug-Excipient Compatibility Studies

S. No	Ingredients	Ratio	Description
S. No	Ingredients	Ratio	Initial	55°C (2 weeks)	40±2°C/70±5%RH (4 weeks)
1	API + Lactose monohydrate	1:5	Off white	No change	No change
2	API + Micro crystalline cellulose	1:5	Off white	No change	No change
3	API + Cross carmellose sodium	1:1	Off white	No change	No change
4	API + Sodium Starch glycolate	1:1	Off white	No change	No change
5	API + Crospovidone	1:1	Off white	No change	No change
6	API + Hydroxy propyl cellulose	1:1	Off white	No change	No change
7	API + Magnesium Stearate	1:1	Off white	No change	No change
8	API + Opadry white	1:1	White	No change	No change

Table 3: Various pre compression parameters of the Formulation blends

Formulation code	Bulk density (gm/ml)	Tapped density (gm/ml)	Compressibility index (%)	Hausner’s Ratio
F1	0.484±0.002	0.547±0.006	11.51±1.89	1.13±0.87
F2	0.478±0.007	0.563±0.004	15.01±1.76	1.17±0.54
F3	0.472±0.003	0.528±0.007	10.6±1.54	1.11±0.64
F4	0.463±0.004	0.556±0.003	16.72±1.34	1.20±0.33
F5	0.456±0.002	0.574±0.002	20.05±1.66	1.24±0.26
F6	0.461±0.005	0.587±0.005	21.46±1.27	1.27±0.61
F7	0.447±0.006	0.593±0.006	24.62±1.65	1.32±0.88
F8	0.477±0.002	0.648±0.007	26.38±1.58	1.35±0.92
F9	0.468±0.002	0.604±0.003	22.51±1.46	1.29±0.51

Figure 1: Calibration curve of Dasatinib in Acetate buffer

Table 4: Various post compression parameters of the Formulations

Formulation Code	Weight Variation	Hardness (kg/cm²)	Friability (%)	Disintegration time (mins)
F1	548.6±2.47	7.1±0.13	0.13±0.03	4±0.16
F2	546.9±3.02	6.3±0.22	0.15±0.01	3.2±0.15
F3	549.4±4.21	7±0.14	0.12±0.02	2.5±0.09
F4	552.6±4.11	6.6±0.21	0.25±0.00	5±0.12
F5	549.2±2.35	7±0.30	0.19±0.02	4.7±0.13
F6	554.1±3.09	7.2±0.11	0.33±0.03	4±0.12
F7	545.5±5.34	7.5±0.13	0.17± 0.02	5.8±0.16
F8	549.4±4.08	6.8±0.24	0.22±0.04	5.2±0.15
F9	543.7±2.22	6.5±0.32	0.37±0.03	4.8±0.14

Table 5: Acclerated Stability studies on the Formulation F3.

S. No	Parameters		Specifications	Test Condition (Accelerated)
				40 ± 2⁰C and 75 ± 5% RH
				0 Day	1 Month	2 Month	3 Month
1	Description		White to off-white round shape film coated tablet.	Comply	Comply	Comply	Comply
2	Moisture content		Not more than 6.0%	4.427%	4.326%	4.211%	4.143
3	Assay		NLS 90% and NMT 110% of labelled amount of drug.	97.9%	98.2%	98.8%	99.6%
4	Related substances by HPLC	Impurity A	NMT 0.2%	ND	ND	ND	ND
		Impurity B	NMT 0.2%	0.006%	0.007%	0.007%	0.007%
		Single max impurity	NMT 0.5%	0.059%	0.064%	0.067%	0.072%
		Total impurity	NMT 2%	0.065%	0.071%	0.074%	0.079%
5	Dissolution		NLT 80% of labeled amount of DST dissolved in 30 min	97.8%	97.2%	96.2%	96.1%

CONCLUSION:

The present investigation was undertaken to formulate and evaluate immediate release Dasatinib film coated tablets for the treatment of lymphoid blast phase chronic myloid leukaemia. Based on the above results, the optimized formulation was the formulation containing Crosscarmellose sodium at 4.5%. Comparison of drug release between the innovator (Sprycel) and the optimized formulation showed that the optimized formulation was capable of giving a higher drug release within the same specified time interval. The optimized formulation was subjected to stability studies and it was found to be stable.

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Received on 07.10.2018 Modified on 07.11.2018

Research J. Pharm. and Tech 2019; 12(2):729-734.

DOI: 10.5958/0974-360X.2019.00129.X