dESIGN AND EVALUATION OF FLUCONAZOLE NIOSOMAL GEL FOR TOPICAL APPLICATION

M.PHARM DISSERTATION PROTOCOL

SUBMITTED TO THE

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA.

By

CHANDAK CHETAN G

Under the guidance of

S.B SHIRSAND

M. Pharm, (Ph.D)

DEPARTMENT OF PHARMACEUTICAL

TECHNOLOGY

H.K.E.S’s COLLEGE OF PHARMACY

GULBARGA-585105

2010-11
Rajiv Gandhi University of Health Sciences, Karnataka Bangalore

Annexure – II

Proforma for Registration of Subject for Dissertation

1. / Name of the candidate
(In block letters) / CHANDAK CHETAN G
Permanent Address / 1707, sadashiv peth opp to scout ground near udyanprasad karyalay, pune 411030
2. / Name of the Institution / H.K.E. Society’s College of
Pharmacy, Sedam Road,
Gulbarga -585105
3. / Course of study and subjects / M.Pharm
(PHARMACEUTICAL TECHNOLOGY)
4. / Date of admission to the course / 12.07.2010
5. / Title of the Topic / dESIGN AND EVALUATION OF FLUCONAZOLE NIOSOMAL GEL FOR TOPICAL APPLICATION
6. / Brief Resume of the Intended work
6.1 / Need for the study:
Fluconazole is an imidazole derivative with broad spectrum antimycotic activity. It acts by inhibiting biosynthesis of sterol ergosterol, an important component of fungal cell membranes. It is widely used for the treatment of local oropharyngeal candidiasis, vaginal yeast infections; topical applications include fungal infections such as ring worm, athlete’s foot and jock itch. When applied topically Fluconazole penetrates the epidermis but there is little if any systemic circulation.[1]
Fluconazole is a synthetic antifungal agent of imidazole class, It act by interacting with 14-α demethaylase, Acytochrome P-450 enzyme necessary to convert lanosterol to ergosterol, as ergosterol is an essential component of the fungal cell membrane. Inhibition of its synthesis results in increased cellular permeability causing leakage of cellular content. Fluconazole also inhibits endogenious respiration and also interact with membrane phospholipids that leads to inhibition of transformation of yeasts to mycelia forms and also inhibits purine uptake, and impairs triglycerides and or phospholipid biosynthesis.[2]
Nowadays, niosomes play an increasingly important role in drug delivery as they can reduce toxicity and modify drug pharmacokinetics and bioavailability. Niosomes seem an interesting drug delivery system in the treatment of dermatological disorders. In fact, topically applied niosomes can increase the residence time of drugs in the stratum corneum and epidermis, while reducing the systemic absorption of the drug. Niosomes have also been widely studied as drug carriers for controlled and targeted delivery.[3,4] Preliminary studies indicate that niosomes behave in-vivo like liposomes, prolonging the circulation of entrapped drug to alter its organ distribution and metabolic stability, or prolonging contact time of drug with the applied tissues in topical application.[5-8] Niosomes are unilamellar or multilamellar vesicles formed from synthetic non-ionic surfactants, offering an alternative to liposomes as drug carriers.
The proposed work investigates the feasibility of niosomes of fluconazole for topical application. To enhance the stability and increase the viscosity of this system, carbapol gel will be prepared.
6.2 / Review of Literature
Exhaustive literature survey was carried out on the proposed topic using Internet, RGUHS(Helinet Consortium) and referring journals. The survey reveals that no work has been carried out on the proposed topic and some related work is quoted below.
Katare et alhave prepared and evaluated (in-vitro) Dithranol liposomal/niosomal delivery system. The amount of drug loaded into these vesicles was in the range of 0.97mg/70mg to 3.51mg/180mg of total lipid. These systems have been found to be reasonably well in their size and stability characteristics and exhibited improved permeation properties. The results reveal the merits of developed drug-loaded liposomes and niosomes and justify their potential in strengthening the efficacy and safety of the drug.[9]
Lithy et al have prepared and evaluated rheological, stability studies of cutina lipogels and gel microemulsion of fluconazole, and also studied the statistical analysis of viscosity data before and after storage under freezing condition using ANOVA test at a level of significance of 0.05. This revealed non-significant difference in viscosity measurement before and after storage for all gels except of cutina CPA lipogel.[10]
Fadd.et alhave studied that in the presence of cholesterol, all amphiphiles used during this study were able to form stable vesicle dispersion with or without tretinoin, during this study, he also prepared niosomes made of span 80, but the vesicular dispersion showed a very low stability and a high TRA leakage.[11]
Bouwsrra have investigated worked on skin structure and mode of action of vesicles, one of the approaches is the application containing vesicles, in order to unveil the mechanisms involved in increasing the drug transport across the skin. And also studied a possible interaction between vesicles and skin.[12]
Kaur et alhave made an approach to study vesicular system in ocular drug delivery .This article reviews the constrain with conventional ocular therapy and explores various novel approaches, in general to improve ocular bioavailability of the drugs.[13]
Jain et alhaveprepared a transdermal delivery system of an analgesic agent using elastic liposomes. Common anti-inflammatory non-steroidal drug diclofenac was chosen to provide sustained and
targeted delivery. Elastic liposomes were prepared and characterized
in-vitroand in-vivo. [14]
Holler etalhave extensively investigated the effect of selected fluorinated drugs in a “ringing” gel on rheological behavior and skin permeation. A transparent ringing gel consisting of surfactants, paraffin oil and water has been used as vehicle in order to investigate in-vitro skin permeation studies, chemical stability and rheology behaviour of selected different fluorinated drugs flucanazole, Flucodrisone-acetate, Flumethasone –Pivalate, Flutanide and Flufenamic acid.[15]
Khazaeli et alcharacterized and studied in-vitro release of Caffeine loaded niosomes.The in-vitro permeation of caffeine from niosomes of various compositions reveals that neutral vesicles prepared with span60 and cholesterol, entrap a higher caffeine amount, at pH 7.4, niosomes may be a promising carrier for caffeine, especially to their simple production and facile scale up.[16]
Mahmoud Mokhtar et alworked on effect of some formulation parameters on Flubriprofen encapsulation and release rate of niosomes prepared from proniosomes with various grades of spanswith and without cholesterol and successively prepared niosomes which suggest the potentiality of proniosomes as stable precursor for the immediate preparation of niosomal carrier systems.[17]
Prabhager Balkrishnan et al have prepared niosomes incorporated Minoxidil and assessed for in-vitro parameters which revealed the potentiality of niosomes in enhancing transdermal bioavailability of Minoxidil in hairless mouse skin even at very low drug concentration unlike control formulations.[18]
Bachhav et al have prepared fluconazole microemulasion based gel which was successfully formulated for the topical treatment of the vaginal candidiasis and was evaluated for in-vitro and in-vivo drug release which gave optimistic results.[19]
Panyosak et al havemade an approach to asses safety of Azelaic acid and its derivatives entrapped in nanovesicles. This study has
demonstrated the safety of Azelaic Acid and its derivatives when entrapped in nanovesicles because of no toxicity to normal cell lines
and no allergy on rabbit skin.[20]
Manosroi et al have evaluated transdermal absorption enhancement through rat skin of gallidermin loaded niosomes and reported that when gallidermin was loaded in anionic niosomes, its chemical degradation at high temperature was not only protected, but also sustained the release effects.[21]
Rathod S, and Deshpande SG, have designed and evaluated Pilocarpine Nitrate liposomal formulation by thin layer film hydration method using Egg Phosphatidycholine andcholesterol where used to make multilamellar vesicles. Liposomes in size range 0.2 to 1 μm were obtained by optimizing the process. Encapsulation efficiency of neutral, positive and negatively charged liposomes were found to be 32.5, 35.4 and 34.2 percent respectively.[22]
6.3 / Objectives of the study
In the proposed research work Fluconazole Niosomes will be prepared with the following objectives.
  1. To bypass the hepatic metabolism of the drug.
  2. To increase the residence time of drug at the target tissue.
  3. Increased skin penetration of the drug.
  4. To prepare more efficient drug delivery system.

7. /

Materials and Methods

7.1 / Materials:
Drug: Fluconazole
Excipients:
  1. Sorbitan Monoesters
  2. Cholesterol
  3. Diethyl Ether / chloroform
  4. Carbopol
  5. Glycerin.etc.

Equipments:
  1. UV-VIS spectrophotometer.
  2. Optical microscope.
  3. Rotary Flash Evaporator (superfit model).
  4. Ultra sonicator.
  5. IR spectrophotometer.
  6. Stability Chamber.
  7. Hot air oven.
  8. Thermostatic hot plate with magnetic stirrer.
  9. Digital overhead stirrer.
  10. Electronic Balance.
  11. Digital pH meter etc.

7.2 / Method
Niosomes incorporated with Fluconazole will be prepared by thin lipid evaporation method; the prepared Fluconazole niosomes are incorporated in carbopol gel.[23]
Evaluation of Fluconazole Niosomes includes:
  1. Size, Shape, Entrapment Efficiency[9]
  2. In-vitrodrug release (exhaustive dialysis method)[9]
  3. In-vitroantifungal activity by cup-plate method
  4. Drug leakage on storage and stability[9]

7.3 / Does the study require any investigation or intervention to be conducted on patients or other humans or animals? If so please describe briefly
Not under the plan of work
7.4 / Has ethical clearance have been obtained from your institution in case of 7.3?
Not applicable
8. / List of references:
1.Sweetman SC. The editor Martindale: The complete drug reference. 35th edn London: Pharmaceutical press. p. 479-81.
2.Fluconazole.[cited 2010 24 Nov]; Available from:
3.Uchegbu IF, Double JA, Turton JA, Florence AT. Distribution, metabolism and tumoricidal activity of doxorubicin administered in sorbitan monostearate (Span 60) niosomes in the mouse. Pharm Res. 1995;12(7):1019-24.
4.Varshosaz J, Pardakhty A, Hajhashemi VI, Najafabadi AR. Development and physical characterization of sorbitan monoester niosomes for insulin oral delivery. Drug Del. 2003;10(4):251-62.
5.Fang JY, Hong CT, Chiu WT, Wang YY. Effect of liposomes and niosomes on skin permeation of enoxacin. Int J Pharm. 2001;219(1-2):61-72.
6.Perini G, Saettone MF, Carafa M, Santucci E, Alhaique F. Niosomes as carriers for ophthalmic drugs: in-vitro/in-vivo evaluation. Boll Chim Farm. 1996;135(2):145-6.
7.Shahiwala A, Misra A. Studies in topical application of niosomally entrapped Nimesulide. J Pharm Pharm Sci. 2002;5(3):220-5.
8.Vora B, Khopade AJ, Jain NK. Proniosome based transdermal delivery of levonorgestrel for effective contraception. J Control Release. 1998;54(2):149-65.
9.Agarwal R, Katare OP, Vyas SP. Preparation and in-vitro evaluation of liposomal/niosomal delivery systems for antipsoriatic drug dithranol. Int J Pharm. 2001;228(1-2):43-52.
10 El Laithy HM, El-Shaboury KM. The development of Cutina lipogels and gel microemulsion for topical administration of fluconazole. AAPS PharmSciTech. 2002;3(4):E35.
11.Manconi M, Sinico C, Valenti D, Loy G, Fadda AM. Niosomes as carriers for tretinoin. I. Preparation and properties. Int J Pharm. 2002;234(1-2):237-48.
12.Bouwstra JA, Honeywell-Nguyen PL. Skin structure and mode of action of vesicles. Adv Drug Deliv Rev. 2002;54(1):S41-55.
13.Kaur IP, Garg A, Singla AK, Aggarwal D. Vesicular systems in ocular drug delivery: an overview. Int J Pharm. 2004;269(1):1-14.
14.Jain S, Jain N, Bhadra D, Tiwary AK, Jain NK. Transdermal delivery of an analgesic agent using elastic liposomes: preparation, characterization and performance evaluation. Curr Drug Del. 2005;2(3):223-33.
15.Holler S, Valenta C. Effect of selected fluorinated drugs in a "ringing" gel on rheological behaviour and skin permeation. Eur J Pharm Biopharm. 2007;66(1):120-6.
16.Khazaeli P, Pardakhty A, Shoorabi H. Caffeine-loaded niosomes: characterization and in-vitro release studies. Drug Del. 2007;14(7):447-52.
17.Mokhtar M, Sammour OA, Hammad MA, Megrab NA. Effect of some formulation parameters on flurbiprofen encapsulation and release rates of niosomes prepared from proniosomes. Int J Pharm. 2008;361(1-2):104-11.
18.Balakrishnan P, Shanmugam S, Lee WS, Lee WM, Kim JO, Oh DH, et al. Formulation and in-vitro assessment of minoxidil niosomes for enhanced skin delivery. Int J Pharm. 2009;377(1-2):1-8.
19.Bachhav YG, Patravale VB. Microemulsion based vaginal gel of fluconazole: formulation, in-vitro and in-vivo evaluation. Int J Pharm. 2009;365(1-2):175-9.
20.Panyosak A, Manosroi J, Rojanasakul Y, Manosroi A. Safety assessment of azelaic acid and its derivatives entrapped in nanovesicles. Hum Exp Toxicol. 2009;28(6-7):387-92.
21.Manosroi A, Khanrin P, Lohcharoenkal W, Werner RG, Gotz F, Manosroi W, et al. Transdermal absorption enhancement through rat skin of gallidermin loaded in niosomes. Int J Pharm.2010;392(1-2):304-10.
22.Rathod S, Deshpande SG. Design and evaluation of liposomal formulation of pilocarpine nitrate. Indian J Pharm Sci. 2010;72(2):155-60.
23.Manosroi A, Jantrawut P, Manosroi J. Anti-inflammatory activity of gel containing novel elastic niosomes entrapped with diclofenac diethylammonium. Int J Pharm. 2008;360(1-2):156-63.
9 / Signature of the Candidate / CHANDAK CHETAN G
10 / Remarks of the Guide / The proposed research topic is novel one, with the preparation of Niosomal Gel we can improve the efficiency of Fluconazole for Fungal Infections
11 / Name and Designation of
11.1 / Guide / S.B SHIRSANDM.Pharm (Ph.D)
Assistant Professor
Department of Pharmaceutical Technology
H.K.E.S’s College of Pharmacy
GULBARGA- 585105
11.2 / Signature
11.3 / Co-Guide / Dr. S.S BUSHETTIM.Pharm, Ph.D
Professor
Department of Pharmaceutical Technology
H.K.E.S’s College of Pharmacy
Gulbarga
11.4 / Signature
11.5 / Head of the Department / Dr. P.V SWAMYM.Pharm, Ph.D
Professor
Department of Pharmaceutical Technology
H.K.E.S’s College of Pharmacy
Gulbarga
11.6 / Signature
12 / Remarks of the Chairman and Principal
12.1 / Signature

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