“DESIGN AND CHARACTERISTION OF NANOEMULSION BASED FLURBIPROFEN GEL FOR TOPICAL DELIVERY”
M. Pharm. Dissertation Protocol
Submitted to the
Rajiv Gandhi University of Health Sciences, Karnataka.
Bangalore
By
DHARMENDRA CHAUDHARY
B. Pharm.
Under the Guidance of
Mr. SARFARAZ Md.
M. Pharm.
Asst. Professor
Department of Pharmaceutics
DEPARTMENT OF PHARMACEUTICS
N.E.T. PHARMACY COLLEGE,
RAICHUR
2013
Rajiv Gandhi University of Health Sciences, Karnataka
Bangalore
ANNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1 / Name of candidate and address (In Block Letters) / MR.DHARMENDRA CHAUDHARYS/O-RAM KARAN CHAUDHARY,
JABDI-2
DIST- SARLAHI, NEPAL
2 / Name of the Institute / N.E.T.PHARMACY COLLEGE, RAICHUR.
3 / Course of study and subject: / M.PHARM. PHARMACEUTICS.
4 / Date of admission of course: / 26-12-2012
5 / Title of the topic:
“DESIGN AND CHARACTERISTION OF NANOEMULSION BASED FLURBIPROFEN GEL FOR TOPICAL DELIVERY”
6 / Brief Resume of this intended work:
6.1 Need for the study Enclosure-I
6.2 Review of Literature Enclosure-II
6.3 Objectives of study Enclosure-III
7 / Materials and Methods:
7.1 Source of data Enclosure-IV
7.2 Method of collection of data (Including sampling procedure, if any)
Enclosure-V
7.3 Does the study require any investigation or interventions to be conducted on patients of humans or animals? If so, please describe briefly.
Yes
7.4 Has ethical clearance been obtained from your institution in case of 7.3?
Yes: IAEC NO: 576/2002/bc/IAEC/CPSCEA
8 / List of References Enclosure-VI
9 / Signature of the candidate
10 / Remarks of the Guide / The proposed work can be carried out in the laboratory.
11 / Name and designation of (in block letters)
11.1 Guide
11.2 Signature /
Mr. SARFARAZ Md.
Assistant Professor
Dept. of Pharmaceutics,
N.E.T. Pharmacy college,
RAICHUR- 584103.
11.3 Co-Guide (if any)
11.4 Signature / ------
------
11.5 Head of Department
11.6 Signature /
Dr. H. DODDAYYA
Professor
Dept. of Pharmaceutics.
.
12 /
12.1 Remarks of the Chairman and Principal
12.2 Signature / Forwarded for scrutinyDr. H. DODDAYYA
Principal,
N. E. T. Pharmacy College,
RAICHUR-584103.
Enclosure-I
6) Brief resume of the intended work.
6.1) Need for the study:
Flurbiprofen a propionic acid derivative is an NSAID (non-steroidal anti-inflammatory drug). It is used in musculoskeletal system and joint disorders such as ankylosing spondylitis, osteoarthritis, rheumatoid arthritis, in soft-tissue injuries such as sprains and strains, for postoperative pain, in mild to moderate pain including dysmenorrhoea and migraine. Flurbiprofen is also used as a lozenge in symptomatic relief of sore throats. Flurbiprofen sodium is used in eye drops to inhibit intra-operative miosis and to control postoperative inflammation of the anterior segment of the eye.
Flurbiprofen is practically insoluble in water, freely soluble in alcohol and in dichloromethane; dissolves in aqueous solution of alkali hydroxides and carbonates. Flurbiprofen sodium is sparingly soluble in water1. Poorly water soluble drugs are associated with slow drug absorption leading eventually to inadequate and variable bioavaibility2.
Oral administration of flurbiprofen is associated with severe gastrointestinal adverse events including abdominal discomfort, constipation, diarrhea, dyspepsia, nausea and vomiting; these are usually mild and reversible but in some patient’s peptic ulceration and severe gastrointestinal bleeding may occur. Moreover, it requires frequent dosing due to its short elimination half-life of 3.9 hr. These facts raise a need for a long-term sustained delivery of flurbiprofen1, 3, 4.
The transdermal route of administration is recognised as one of the potential route for local and systemic delivery of drugs. Transdermal delivery not only provides controlled, constant administration of the drug but also allows continuous input of drug with short biological half life, reduced frequency of administration, reduced side effects, reduced load on digestive tract and liver that oral route places and improved patient compliance5.
Many of the dermal vehicles contain chemical enhancers and solvents to modify drug permeation through skin. But use of these chemical enhancers may be harmful, especially in chronic application, as many of them are irritants. Therefore, it is desirable to develop a topical vehicle system that does not require the use of chemical enhancers to facilitate drug permeation through the skin. One of the most promising techniques for enhancement of skin permeation of drugs is nanoemulsion6. Recently, increasing attention has been focused on nanoemulsion based drug delivery system due to their ease of formulation with biocompatible excipients, and unique properties such as smaller droplet size, increasing solubility and dissolution rate, improving diffusion and mucosal permeability7. Nanoemulsions offer several significant advantages including low skin irritation, powerful permeation ability, and high drug-loading capacity for topical delivery when compared with the other carriers such as liposome or solid lipid nanoparticles8.
Nanoemulsions are quaternary system composed of an oil phase, a water phase, surfactant frequently in combination with co-surfactant. These spontaneously formed systems possess precise physiochemical properties such as transparency, optical isotropy, low viscosity and thermodynamic stability. In stable emulsion, droplet diameter is usually within range of 10-100nm (100-1000 A°)9. They are kinetically stable without any apparent flocculation or coalescence during the long term storage due to their nanometre sized droplets10, 11. The utility of a nanoemulsion has been successfully established in optimising the therapeutic performance of many lipophilic drugs which further justifies rationale of the study12. The high solubilising capacity of a nanoemulsion enables them to increase the solubility of poorly water soluble drugs. Both increase in solute concentration and tendency of drug to favour portioning into stratum corneum makes nanoemulsion a useful vehicle to enhance transdermal drug permeability13.
Hence an attempt is made to prepare nanoemulsion based gel of flurbiprofen that can increase the solubility as well as permeability of the drug.
Enclosure-II
6.2) Review of literature:
Ø Balakrishnan P et. al., studied the effect of process parameters on nanoemulsion droplet size and distribution in SPG (Shirasu-porous-glass) membrane emulsification. A SPG membrane with a mean pore size of 2.5 µm was used to produce an O/W nanoemulsion of flurbiprofen consisting of methylene chloride as dispersed phase, polyvinyl alcohol (PVA) as the stabilizer and a mixture of Tween 20 and Tween 80 in demineralized water as the continuous phase. Increasing the agitator speed and stabilizer concentration increased the z-average diameter and decreased the size uniformity. There was a linear relationship between the increased feed pressure and decreased z-average diameter of emulsion droplets. The nanoemulsion composed of oil, water, PVA and the surfactant mixture at the weight ratio of 10/100/1/514.
Ø Jin-qiu et. al., prepared flurbiprofen axetil nanoemulsion-in situ gel system (FBA/NE-ISG) and observed its ocular pharmacokinetics, rheological behavior, TEM images, irritation and cornea retention. Nanoemulsion was prepared by high-speed shear and homogenization process, and then mixed with gellan gum to prepare FBA/NE-ISG. Rheological study showed that FBA/NE-ISG possesses strong gelation capacity; TEM images suggested no significant change in particle morphology of pre and post gelation. Good ocular compatibility of FBA/NE-ISG was testified by the irritation test based on histological examination. In-vivo fluorescence imaging system was applied to investigate the characteristics of cornea retention, and the results indicated that the nanoemulsion in-situ gel (NE-ISG) prolonged the cornea retention time significantly. Pharmacokinetics of FBA/NE-ISG in rabbit aqueous humor showed that the ocular bioavailability was improved greatly by the novel preparation15.
Ø Baboota S et. al., investigated the potential of nanoemulsion formulation for transdermal delivery of celecoxib (CXB). The in-vitro skin permeation profile of optimized formulations compared with CXB gel and nanoemulsion gel showed significant increase in the steady state flux (Jss), permeability coefficient (Kp) and enhancement ratio (Er).The highest value of permeability parameters was obtained in formulation consisting of 2% (m/m) of CXB, 10% (m/m) of oil phase (Sefsol 218 and Triacetin), 50% (m/m) of surfactant mixture (Tween-80 and Transcutol-P) and 40% (m/m) water. The anti-inflammatory effects of formulation showed a significant increase (p < 0.05) in inhibition after 24 hr compared to CXB gel and nanoemulsion gel on carrageenan-induced paw edema in rats. These results suggested that nanoemulsions are potential vehicles for improved transdermal delivery of CXB6.
Ø Modi JD et. al., investigated the potential of a nanoemulsion formulation for topical delivery of aceclofenac. The nanoemulsion area was identified by constructing psedoternary phase diagram. The in-vitro skin profile of optimised formulation was compared with that of aceclofenac conventional gel and nanoemulsion gel. A significant increase in permeability parameters such as steady-state flux (Jss), permeability coefficient (Kp), and enhancement ratio (Er) was observed in optimised nanoemulsion formulation consist of 2% w/w of aceclofenac, 10% w/w of Labrafac, 45% w/w surfactant mixture (Cremophor EL: Ethanol), and 43% w/w of distilled water. The anti-inflammatory effects of formulation showed a significant increase in percent inhibition value after 24 hr when compared with aceclofenac conventional gel and nanoemulsion gel on carrageenan-induced paw edema in rats. These results suggested that nanoemulsions are potential vehicles for transdermal delivery of aceclofenac16.
Ø Wais M et. al., investigated the nanoemulsion system for enhanced percutaneous penetration of glibenclamide (GLBD). Pseudoternary phase diagrams were constructed in order to optimize the surfactant, co-surfactant and surfactant: co-surfactant ratios (Smix). The nanoemulsion formulation consisted of Labrac and Triacetin (1:1) as an internal oil phase in external aqueous phase, Tween 80 as surfactant and diethylene glycol monoethyl ether as co-surfactant. Pseudoternary phase diagram was developed to determine the effect of the surfactant to co-surfactant ratio (Smix) on the emulsion formation, a transparent region. The nanoemulsion formulation had small droplet size (<117 nm), uniform size distribution (PI<0.247), and low viscosity (<73.0 mP). Novel GLBD formulation could be designed and projected to be suitable for transdermal application9.
Ø Idrees MA et. al., prepared flurbiprofen microemulsion for enhanced transdermal delivery and studied effects of different surfactants and co-surfactants on its phase behaviour. Various surfactant and co-surfactant mixtures in ratio of 2:1 (Smix) along with oleic acid (oil) were selected and phase diagrams were constructed. In vitro transdermal permeation of all microemulsion was high than saturated aqueous drug solution. The type of surfactant did not affect the droplet size, propylene glycol as co-surfactant produced the largest droplet and highest viscosity. Decrease in oil or Smix concentration resulted in decrease of the droplet size and increase in permeation flux, while decrease in viscosity also increased the permeation flux of microemulsion. Finally the selected formulation comprising 5% flurbiprofen, 5% oleic acid, 46% Tween 20: ethanol (2:1) and 44% water showed the highest transdermal flux and caused no irritation17.
Ø Tirunagari M et. al., investigated the effect of β-cyclodextrin (β-CD) and co-solvents on the solubility of flurbiprofen. The increase in solubility with 1 mM, 2 mM, and 3 mM was found to be 7.02 times, 11.51 times and 11.41 times respectively from the phase solubility diagram. The drug solubility was increased linearly up to 2 mM of β-CD followed by hardly increasing the solubility. Later, the solubility of flurbiprofen in three different co-solvents polyethylene glycol 400 (PEG 400), propylene glycol and ethanol was determined and found to be 7.38 times, 19.43 times, and 12.34 times. From the above results the optimum concentration of β-CD (2 mM) and one of the best co-solvent (propylene glycol) was selected and the effect on increase in solubility by the combination was found to be 18.85 times. It was concluded that co-solvent propylene glycol was the best solubiliser among all selected18.
Enclosure-III
6.3) Objectives of the study:
The present study is planned with the following objectives:
1) To perform preformulation studies on drug.
2) To screen out surfactant, co-surfactant and construct the pseudoternary phase diagram to identify nanoemulsion region.
3) To formulate nanoemulsion based gel by suitable methods.
4) To study drug and excipient interaction by using FT-IR and DSC instruments.
5) To characterize various physiochemical parameters for developed nanoemulsion gel.
6) To evaluate the nanoemulsion gel for in-vitro diffusion studies using Franz diffusion cell or Keshari diffusion cell using artificial or biological membrane.
7) To perform in-vitro permeation studies on best formulations using rat abdominal skin.
8) To perform in-vivo studies on rat to compare anti-inflammatory effect of best formulation with marketed formulation.
9) To perform the stability studies for selected formulations as per ICH guidelines.
Enclosure-IV
7) Materials and Methods:
7.1) Source of data:
Primary data: This data will be collected by conducting laboratory experiment and recording the observation.
Secondary data: This data will be collected from:
Internet.
Review articles.
Research publications.
International and Indian journals. •Textbooks and reference books
Enclosure-V
7.2) Method of collection of data:
The data for the study is planned to collect from the laboratory-based experiments:
1. Preformulation studies like solubility, melting point and characterization of the drug and excipients will be done by employing suitable methods. Compatibility of drug with excipients is analyzed by using FT-Infra-Red Spectroscopy and Differential Scanning Calorimetry instruments adopting reported methods.
2. Preparation of flurbiprofen nanoemulsion based gel by spontaneous nanoemulsification or any other suitable methods.
3. Characterization of developed nanoemulsion based gel for:
a. Physicochemical properties on nanoemulsions:
i. Droplet size and Size distribution
ii. Viscosity
iii. Refractive index,
iv. pH
v. TEM photomicrographs
b. Total drug content by UV spectrophotometer.
c. Zeta potential using Zeta meter or any other suitable methods.
d. In-vitro drug release study using Franz diffusion cell with artificial or biological membrane, or dialysis bag diffusion method or any other suitable methods.
e. In-vitro permeation studies employing rat abdominal skin.
f. In-vivo studies using rat to perform anti-inflammatory activity.
4. For selected formulations stability studies will be carried out using stability chamber as per ICH guidelines.
STATISTICAL ANALYSIS
All values will be expressed as mean ± SEM for 10 animals in a group. Results will be subjected to statistical analysis using one way ANOVA (analysis of variance) followed by Dunnett’s test, p < 0.05 will be considered as statistically significant.