EFFECT OF DIFFERENT CARRIERS ON IN-VITRO …ijupbs.com/Uploads/3. RPA1415115115.pdf · Poor...

25 | P a g e International Standard Serial Number (ISSN): 2319-8141

Full Text Available On www.ijupbs.com

International Journal of Universal Pharmacy and Bio Sciences 3(5): September-October 2014

INTERNATIONAL JOURNAL OF UNIVERSAL

PHARMACY AND BIO SCIENCES IMPACT FACTOR 2.093***

ICV 5.13*** Pharmaceutical Sciences RESEARCH ARTICLE……!!!

EFFECT OF DIFFERENT CARRIERS ON IN-VITRO RELEASE OF

LORNOXICAM

Sagar Bhaskar Patil*, Dr. C.S.Magdum, Dr. S.K.Mohite

Rajarambapu College of Pharmacy, Kasegaon, Tal. Walwa, Dist. Sangli.

KEYWORDS:

Lornoxicam,Opoid

analgesic, NSAID.

For Correspondence:

Sagar Bhaskar Patil*

Address:

Rajarambapu College of

Pharmacy, Kasegaon,

Tal. Walwa, Dist. Sangli.

E-mail:

[email protected]

om

ABSTRACT

Lornoxicam (LOR) is a NSAID of the oxicam class with

analgesic, anti-inflammatory and antipyretic properties. It is a

potent inhibitor of both COX-1 and COX-2 enzymes . It is

available only in tablet and parenteral forms, therefore, the

objective of this project is to study effect of different carriers on

in vitro permeation of LOR in gel forms for topical application.

Since LOR is a lipophilic drug, having a short plasma elimination

half-life, small dose and is a potent drug, it can be considered as a

good candidate for topical application.

mailto:[email protected]

mailto:[email protected]



INTRODUCTION:

Poor dissolution characteristics of relatively insoluble drugs have long been a problem to

pharmaceutical industry. No . of modern drugs are poorly soluble in water and aqueous fluids.

Their absorption and bioavailability require improvement in the dissolution rate and

bioavailability, cyclodextrin (CD) complexation was found to be very successful with a number

of poorly soluble drugs1.Cyclodextrin have mainly been used as complexing agent to increase

the aqueous solubility of poorly water-soluble drugs and to increase their bioavailability and

stabality2

.The most common cyclodextrin are α- cyclodextrin, β- cyclodextrin, and γ

cyclodextrin, which consist of six, seven, and eight glucopyranose units, respectively. Among α-,

β-,γ-;β-CD was used for the study, as it has bigger cavity size and is the least toxic among the

other natural cyclodextrin3.Most of Nonsteroidal anti inflammatory drugs(NSAID) belong to

class II category under Biopharmaceutical classification system cyclodextrin (BCSII) i.e., they

are inherently highly permeable through biological membranes, but exhibit low aqueous

solubility.They need enhancement in solubility and dissolution rate for improving their

bioavailability in the present investigation studies were carried on effect of different carriers on

in vitro permeation of NSAID .Lornoxicam which chemically (3E)-6-chloro-3-[hydroxy(pyridin-

2-ylamino)methylene]-2-methyl-2,3-dihydro-4H- thieno[2,3-e][1,2]thiazin-4-one 1,1-dioxide.

Lornoxicam is a non-steroidal anti-inflammatory drug (NSAID) that belongs to the oxicam class.

As with other NSAIDS, lornoxicam is a potent inhibitor of the cyclooxgenase enzymes, which

are responsible for catalyzing the formation of prostaglandins (act as messenger molecules in the

process of inflammation) and thromboxane from arachidonic acid. Unlike some NSAIDS,

lornoxicam's inhibition of cyclooxygenase does not lead to an increase in leukotriene formation,

meaning that arachidonic acid is not moved to the 5-lipoxygenase cascade, resulting in the

minimization of the risk of adverse events. Like other NSAIDS, lornoxicam's anti-inflammatory

and analgesic activity is related to its inhibitory action on prostaglandin and thromboxane

synthesis through the inhibition of both COX-1 and COX-2. This leads to the reduction of

inflammation, pain, fever, and swelling, which are mediated by prostaglandins. However, the

exact mechanism of lornoxicam, like that of the other NSAIDs, has not been fully determined.

Lornoxicam is used for the treatment of various types of pain, especially resulting from

inflammatory diseases of the joints, osteoarthritis, surgery, sciatica, and other inflammations.

The drug is contraindicated in patients that must not take other NSAIDs, possible reasons

including salicylate sensitivity, gastrointestinal bleeding and bleeding disorders, and severe

impairment of heart, liver or kidney function. Lornoxicam is not recommended during pregnancy



and breastfeeding and is contraindicated during the last third of pregnancy. Lornoxicam has side

effects similar to other NSAIDs, most commonly mild ones like gastrointestinal disorders

(nausea and diarrhea) and headache. Severe but seldom side effects include bleeding,

bronchospasms and the extremely rare Stevens–Johnson syndrome. Interactions with other drugs

are typical of NSAIDs. Combination with vitamin K antagonists like warfarin increases the risk

of bleeding. Combination with ciclosporin can lead to reduced kidney function, and to acute

renal failure in rare cases. Lornoxicam can also increase the adverse effects of lithium,

methotrexate and digoxin and its derivatives. The effect of diuretics, ACE inhibitors and

angiotensin II receptor antagonists can be reduced, but this is only relevant in patients with

special risks like heart failure. As with piroxicam, cimetidine can increase plasma levels but is

unlikely to cause relevant interactions.Lornoxicam has low molecular weight 371.8192 g/mol in

order to avoid irritation of the gastrointestinal tract , minimize systemic toxicity and achieve a

better therapeutic effect, one promising method is to administer the drug via the skin9..However,

the skin is a natural barrier, and only a few drugs can penetrate the skin easily and in sufficient

quantities to be effective.Therfore,in recent years, numerous studies have been conducted in the

area of penetration enhancement9,10.

.Penetration enhancers improve the ability of skin to absorb

drugs by altering the lipid domain of the stratum corneum . The drug used in this work is

Lornoxicam which is a nonsteroidal anti-inflammatory drug.Lornoxicam is used for the

treatment of various type of pain, especially resulting from inflammatory diseases of the joints,

osteoarthritis, surgery , sciatica , and other inflammations. Category of Lornoxicam is

nonselective cox inhibitors (traditional NSAIDS) as Oxicam derivatives. The various research

studies on release properties have been revealed that the active ingredients in gel based

formulations are better percutaneously absorbed than cream or ointment bases.

Materials and Methods:

The materials include Lornoxicam (gift sample from Aristo Pharmaceuticals, Mumbai).Beta

cyclodextrin (gift sample from Loba Chemicals, Mumbai ), HPMC Research Lab, Islampur .

Carbopol 940 Research Lab, Islampur. SLS Research Lab, Islampur. Chloroform Research

Lab, Islampur. Dichloromethane Research Lab, Islampur. DMSO Research Lab, Islampur.

Preparation of Lornoxicam Β-Cyclodextrin Complex Kneading Method:

Calculated amount of Lornoxicam β-CD were accurately weighed and transferred in a glass

morter, Lornoxicam is dissolved in solvent blend of Chloroform : Dichloromethane(1:1volume

ratio),triturate with a small volume of distilled water and solvent blend and kneaded for 60 min

.The product was kept at room temperature for 24 hrs. Distilled water and solvent blend was



used as wetting agent to achieve better interaction the kneaded formulation were prepared at

1:05,1:1,1:1.5,1:2,1:3,and1:4molar ratios. The complex in the ratio 1:2 showed better

complexation.

Preparation of Gel:

Gels were prepared by using different polymers as shown in formulation table. The polymer and

purified water I.P. were taken in a mortar and allow to soaking for 24 hrs. Lornoxicam dissolved

in solvent blend of Chloroform:Dichloromethane and other additives were added .The above

mixture was triturated under homogenizer at constant speed for 2 to 4hrs till the drug dispersion

obtain through the gel.pH of gel was adjusted with triethanolamine when carbopol gels were

prepared .The permeation enhancers like sodium lauryl sulphate and dimethyl sulfoxide were

incorporated in different concentrations. Gel containing Lornoxicam-β-CDcomplex were

prepared by the same procedure as above by adding complex in place of plain Lornoxicam.

Ingredients F1 F2 F3 F4 F5 F6

Lornoxicam(mg) 200 200 200 200

Carbopol940(mg) 0.8 1 1

HPMC(mg) 5 5

β-cyclodextrin(mg) 1:1 1:1.2

Chloroform(ml) 5 5 5 5 5 5

Dichloromethan(ml) 5 5 5 5 5 5

TEA(ml) 0.5 0.5

SLS(mg) 200 250 200 250

Oleic acid(ml) 5

DMSO(ml) 5

Purified water(ML) (q.s. to 100) 100 100 100 100 100 100

Preformulation Studies:

Preformulation studies were performed to determined the physicochemical properties of drug

that could affect the development and efficiency of new drug formulations. Authentifaction of

drug was done by Mr. B.N. Sharma purches officer Aristo Pharmaceuticals Mumbai. The

sample of drug was visually observed for the colour , odour and appearance . Melting point of

Lornoxicam was determined by thiele’s tube method .Excess amount of Lornoxicam was added

to 15 ml of dissolution medium in a 25 ml beaker maintained at 37 ± 0.5 0 C in a constant

temperature water bath for 2h. At appropriate times i.e. 5,10,15,30,45,60,75 min; aliquots of 1ml

were taken from the dissolution medium , filtered and then diluted to 100 ml. Drug contents was

assayed spectrophotometrically at 354 nm. Calibration curves were used for the determination of

the amounts dissolved. UV Spectroscopy Selection of sampling wavelength for analysis λmax

determination using model Jasco V-630. UV-spectrum of pure Lornoxicam was taken in



Chloroform. Drug (10 mg) was dissolved in 100 ml Chloroform to obtain the stock solution of

concentration 100 µg/ml. From this stock solution, 1 ml was withdrawn and diluted to 10 ml.

Absorbance was checked using UV- spectrophotometer. It should give 1 peak corresponding to

its λmax at 354nm. standard stock solution contain Pure Lornoxicam powder equivalent to 100

mg was accurately weighted and dissolved in 100 ml of Chloroform in a 100 ml volumetric

flask and the volume was made up to the mark with 0.1 N Chloroform to obtain final

concentration of 1000 μg/ml ( stock ‘A’ solution ). From the above stock ‘A’ solution 10 ml of

aliquot was pipette out in a 100 ml volumetric flask and the volume was made up to mark with

0.1 N Chloroform to obtain the final concentration of 100 μg/ml ( stock ‘ B ’ solution ) 4 nm.

From the standard stock solution ‘ B ’ various of dilutions ranging between 1-20 μg/ml were

prepared and scanned in the wavelength range of 400-200 nm using UV Spectrophotometer .

Preparation of calibration curve for Lornoxicam in Chloroform include Appropriate aliquots

were pipetted out from the standard stock ‘ B ’ solution into a series 10ml volumetric flask. The

volume was made up of the mark with 0.1 N Chloroform to get a set of solutions having the

concentration range of 2,4,6,8,10,and 12 μg/ml for Lornoxicam.

Absorbance of the above solutions were measured at 354 nm and a calibration curve of

absorbance against concentration was plotted the drug obeys Beer’s law in the concentration

range of 3-18 μg/ml Preparation of calibration curve for Lornoxicam in phosphate buffer pH 7.4

contain Preparation of the standard stock solution ‘ A’ and ‘ B ’ remains same as above from

the standard stock solution ‘ B ’ various dilution ranging between 1-20 μg/ml were prepared and

scanned in the wavelength range of 400-200 nm using Spectrophotometer having the

concentration range of 2,4,6,8,10, μg/ml . Absorbance of the above solution were measured at

354 nm and a calibration curve of absorbance against concentration was plotted.

Fig. no.1: UV spectrum of Lornoxicam



Table no.1: Data for Calibration curve of Lornoxicam in Chloroform

Fig. no.2: Calibration curve of Lornoxicam in chloroform

Table no.2: Data for Calibration curve of Lornoxicam in 7.4 pH Phosphate Buffer

Concentration

( μg / ml )

Absorbance

0 0

2 0.1756

4 0.3189

6 0.4873

8 0.6413

10 0.8003

12 0.9511

Concentration

( μg / ml )

Absorbance

0 0

2 0.0105

4 0.0154

6 0.0204

8 0.0243

10 0.0381



Fig.no.3 Calibration curve of Lornoxicam in artificial Saliva pH 7.

IR spectroscopy:

Infrared spectroscopy is one of the most powerful analytical techniques which offer the

possibility of chemical identification. This technique when coupled with intensity measurements

may be used for quantitative analysis. One of the most important advantages of IR over the other

usual methods of structural analysis is that it provides useful information about the structure of

molecule quickly without tiresome evaluation methods. The technique is based upon the simple

fact that a chemical substance shows marked selective absorption in the IR region. After

absorption of IR radiation the molecules of chemical substance vibrates at many rates of

vibration, giving rise to close packed absorption bands, called IR absorption spectrum which

may extend over wide wavelength range FTIR measurement of the drug was obtained on FTIR

Jasco-410 .Sample was prepared by mixing with KBr (2mg sample in 200 mg KBr ) and placing

in the sample holder . the spectra were scanned over the wave number range of 4000-400 cm-1

at

the ambient temperature . The IR of Lornoxicam and drug polymer combination are given.



Fig. no.4:IR spectra of Lornoxicam

Table No. 3: Interpretation of IR spectra of Lornoxicam

Structural formula of

lornoxicam

Region (in cm-1) Type of Vibration Functional group

3446.17 Stretching OH

3068.19 Stretching N –H

1620.8 Stretching C = C

1646.7 Stretching C = O

1231.33 Aromatic C - H

1146.47 - -SO2

765.60 Stretching C-CL

Fig.no. 5:IR Spectra of Lornoxicam and carbapol:



Fig. No.6: IR spectra of Lornoxicam and Βeta-cyclodextrin

Fig. No.7 IR spectra of Lornoxicam and hpmc

Evaluation of gel:

Determined the pH of the suspension within 5 minutes ,using digital pH meter. A rheological

property of only selected formulations of Lornoxicam was determined. The Spreadability is term

express to denote the extent of area to which the paste readily spreads on application area.

Tube extrudability was then determined by measuring the amount of gel extruded through the tip

when a pressure was apply on tube gel. Gel viscosity measurements were evaluated using a

Brookfield digital viscometer (DV –II Ultras programmable Rheometer, USA) using spindle

no.3 by applying increasing values of the shear rate, in order to reveal possible flow behavior of

the gels. All viscosities measurements were performed at controlled temperature of 30

0c. Drug

content was determined in each of the formulations. About 100mg drug dissolved in 100ml

phosphate buffer of pH 7.4 volumetric flask containing this shake for 2 hours by mechanical

shaker. Solution filter and estimate at UV Spectrophotometer at 354 nm using phosphate buffer

7.4 pH as blank.



The evaluation of percutaneous absorption of drug from Topical semisolid preparation is an

essential requirement for Topical delivery of drug. This can be simplified by monitoring with

two consecutive steps: the first is the release of the drug from the vehicle and the second is its

penetration through the mucus–barrier. These studies provide information about the

physicochemical parameters involved in percutaneous absorption such as the solubility of the

drug in specific vehicle utilized and its diffusion coefficient. In-vitro drug release investigation

serves as an important tool for assessing the various formulations and processing factors during

manufacturing, as a routine quality control measure to assure batch to batch uniformity and

efficacy of the semisolid preparations. In-vitro study stimulates the evaluation of the complex

process involved in drug application, drug release and absorption from the mucus–barrier.

Table no.4: Physical evaluation of Formulations

Formulation No. pH Spredability

(gm.cm/sec)

Viscosity

(cps )

Tube Extrudability Drug Content (% )

F1 7.1 12.8 10538 Good 91.74

F2 7.2 12.4 12735 Good 92.52

F3 6.8 14.8 11220 Good 89.42

F4 6.7 14.6 10954 Good 89.22

F5 6.9 12.5 11599 Good 97.21

F6 7.2 12.3 12878 Good 98.24

Table no. 5: Cumulative % drug release of formulation F1-F6

Time in

min.

Formulation Code

F1 F2 F3 F4 F5 F6

30 40.33 37.85 36.32 38.86 40.81 43.75

60 58.25 43.15 43.75 51.05 56.32 59.68

120 61.82 56.32 61.82 63.27 65.12 61.71

180 68 61.83 70.77 75 70.77 68

240 75 82.83 75 79.18 79.18 74.12

300 80.27 84.12 79.18 86.25 89.22 86.25

360 91.15 89.22 86.27 90.22 92.26 94.25



Fig.no.8: in-vitro drug release profile

RESULT AND DISCUSSION:

The Present study involves the effect of different carriers on in-vitro permeation of Lornoxicam.

The solubility of the drug found to be increased considerably by β Cyclodextrin complexation.

The result were shown in table 4.IR Spectra’s were shown in fig.4-7.The results of in- vitro drug

drug release were indicated graphically in fig. no .8.

CONCLUSION:

The present study revealed that the prepared Lornoxicam Gel formulations with analgesic and

anti-inflammatory properties also useful in arthritis and acute pain after minor and major surgery.

In-vitro drug release studies showed that, the formulation code F6 showed optimum release

(94.25%) in 360 mins. Carbopol 940 & beta cyclodextrin complex 1:1.2 has contributed for

better drug release profile. During our Physicochemical evaluation studies all the formulations

were found to have good tube extrudability, good Spreadability and viscosity characteristics.pH

determination shows that all the formulations have near neutral pH and does not changed during

the study period. IR studies revealed that there is no drug excipient interaction.

REFERENCES:

1. Hsieh D. Drug Permeation Enhancement-Theory and Applications. In: Drug and the

Pharmaceutical Sciences. 4thed. New York: Marcel Dekker; 1987.p.11-13.

2. Rashmi, Garg R, Kumar S, Gupta GD.Topical Gel: A Review. 2008; 6:303-311.

Available from WWW.Pharmainfo.net.

3. Sharma S, Goswami JP, Gupta GD, Mustafa A. Topical Drug Delivery Systems: A

Review. 2009; 6:231-236. Available from WWW.Pharmainfo.net.

http://www.pharmainfo.net/

http://www.pharmainfo.net/



4. Waugh A, Grant A. Ross and Wilson Anatomy and Physiology in Health and Illness. 9th

ed. NewYork: Elsevier Science Ltd.; 2003. p. 361-372.

5. Banker, Rhodes P. Modern Pharmaceutics. 4th ed. New York: Mercel Dekker;

2000.p.189-191.

6. Jain NK.Controlled and Novel Drug Delivery.1st ed. Delhi: CBS Publishers and

Distributors; 1997.p.100-106.

7. Banker GS, Chalmers RK. Pharmaceutics and Pharmacy Practice.1sted. Lippincott

Company; 1982. p.28-294.

8. Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems.5thed. Lippincott

Williams and Wilkins: A. Walters KluwerPublishing Company; p. 307.

9. Beetge, E., du Plessis, J., M¨uller, D.G., Goosen, C., van Rensburg,F.J., 2000. The

influence of the physicochemical characteristics and pharmacokinetics properties of

selected NSAID’s on their transdermal absorption of. Int. J. Pharm. 193, 261–264.

10. Pharmacology and Pharmacotherapeutics, Revised 17th Edition. Satoskr RS, Bhandarkar

SD, Ainapure SS, Popular Prakashan, Mumbai 2001: Page 155-156.

11. Chein YW. Transdermal controlled systemic medications. Pharm. Res. 1988; 5.

12. Tripathi KD. Essentials of medical pharmacology, 6th edition New Delhi: Jaypee

Brothers Medical Publishers (P) Ltd.; 2006: 476-574.

13. Nitalikar M. M.et. formulation and evaluation of topical Gel of Melooxicam with β

cyclodextrin complex Research Journal of Pharmacy Tech., 2013, 6(7);p.790-793.

EFFECT OF DIFFERENT CARRIERS ON IN-VITRO …ijupbs.com/Uploads/3. RPA1415115115.pdf · Poor...

Documents

Transcript of EFFECT OF DIFFERENT CARRIERS ON IN-VITRO …ijupbs.com/Uploads/3. RPA1415115115.pdf · Poor...