PREPARATION AND EVALUATION OF NSAID NANOEMULSION
M.PHARM DISSERTATION PROTOCOL
SUBMITTED TO
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA
BY
ANJANI PRASAD REDDY.VANGA
I M.PHARM
UNDER THE GUIDANCE OF
Dr. B. PRAKASH RAO
Professor
DEPARTMENT OF PHARMACEUTICAL TECHNOLOGY.
KARNATAKA COLLEGE OF PHARMACY
BENGALURU-560064
(2010-2011)
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA, BENGALURU.
ANNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION
1 / Name and Address of Candidate / ANJANI PRASADREDDY. VKarnataka College Of Pharmacy
# 33/2 Thirumenahalli
Hegde Nagar Main Road
Bengaluru-560064.
PERMANENT ADDRESS
S/O V.SUBBAREDDY,
H.No:-3-28-18/18,
Brindavan garden’s , 3rd lane, GUNTUR
Dist: Guntur.
State: Andhra Pradesh-522403.
2 / Name of the Institution / Karnataka college of pharmacy
# 33/2 Thirumenahalli
Hegde Nagar Main Road
Bengaluru-560064.
3 / Course of the Study and Subject / M. PHARMACY
(PHARMACEUTICAL TECHNOLOGY)
4 / Date of Admission / 03-NOV-2010
5 / Title of the Project
PREPARATION AND EVALUATION OF NSAID NANOEMULSION.
6.
6.1 / BRIEF RESUME OF INTENDED WORK
NEED FOR THE STUDY
Nanotechnology comprises technological developments on the nanometer scale, usually 0.1-1000nm. The use of nanotechnology in the field of pharmaceuticals has grown over the last few years. The pharmaceuticals developed based on the nanotechnology are termed as “NANOPHARMACEUTICALS”. The various nanopharmaceuticals currently being used or in the process of development of Nano emulsions, nanosuspensions, nanospheres, nanotubes, nanocells etc.
Nanoemulsions are a group of dispersed particles used for pharmaceutical and biomedical aids and vehicles that show great promise to the future of drug therapies, cosmetics, diagnostics and biotechnologies. Nanoemulsions are defined as the oil-in-water (o/w) emulsions with mean droplet diameters ranging from 50 to 1000nm. Usually, the average droplet diameters ranging from 100 to 500 nm. The particles can exist as water-in-oil and oil-in-water forms, where the core of the particle is either water or oil respectively. Nanoemulsions are made from surfactants approved for human consumption and common food substances that are “Generally recognised as safe” by the FDA. The emulsions are easily produced in large quantites by mixing a water immiscible oil phase into an aqueous phase with a high stress, a mechanical extrusion process that is available worldwide.
The nanoemulsions are also referred as mini emulsions, ultrafine emulsions and sub micron emulsions. Phase behaviour studies have shown that the size of the droplets is governed by the surfactant phase structure at the inversion point induced by either temperature or composition. Studies on the nanoemulsion formation by the phase inversion temperature method have shown a relationship between minimum droplet size and complete solubilisation of the oil in a microemulsion bicontinuos phase independenty of whether the initial phase equilibrium is single or multiphase.
Due to their small droplet size nanoemulsions possess stability against sedimentation or creaming with Ostwald ripening forming the main mechanism of nanoemulsion breakdown.
Non steroidal anti-inflammatory drug (NSAID) has been used in the treatment of various kinds of pains, inflammation, and arthritis.
6.2 / REVIEW OF LITERATURE
· Ganciclovir plays an important role in the treatment of ocular viral infections. The aim of the present study was to investigate the comparative potential of different mucoadhesive nano formulations for the topical ocular delivery of Ganciclovir. Ganclovir mucoadhesive Nanoemulsions were prepared by the reverse-phase evaporation technique. The formulations were evenly round in shape with mean particle size in the range of 23-200 nm. These results indicated the nonirritant and nontoxic nature of the developed formulations. (Akhter S, 2011)
· The plan this work was to develop and characterize semisolid topical formulations containing nimesulide-loaded nanoemulsion. The spontaneous emulsification method was used to prepare the nanoemulsion. The drug-loaded nanoemulsion formulation was incorporated in Carbopol 940 gels. The semisolid dosage forms showed yellowish, glossy and homogeneous aspect after the incorporation of the nanoemulsion. The recovery of nimesulide and the pH values for the gels containing nanoemulsion remained constant during storage (120 days). For the formulation, the rheograms exhibited a non-Newtonian behaviour presenting pseudo plastic characteristics and shear thinning. The rheograms were adjusted to Ostwald's model showing regression coefficients higher than 0.9900. None thixotropic phenomenon was experimentally detected under the test conditions for all formulations. (Alves PM, 2005)
· The aim of this work was to evaluate the in vitro performance of a nebulized nanoemulsion formulation which had been optimised previously. To do so, a transparent nanoemulsion preparation containing 1.5 mg/ml of budesonide was prepared and diluted to achieve concentrations of 250 and 500 μg/ml budesonide. The in vitro characteristics of the diluted nanoemulsions were then compared with the commercially available suspension of budesonide when nebulized using a jet and a vibrating mesh nebulizer. A smaller MMAD with improved aerosol output was observed in the nanoemulsion preparations compared with the corresponding suspension formulations indicating an improved in vitro performance for the nanoemulsion-based preparations. (Amani A, 2010)
· A promising strategy based on the antisense oligonucleotides against the Plasmodium falciparum topoisomerase II has been considered using cationic nanoemulsion as oligonucleotide delivery system. Phosphodiester and chemically modified phosphorothioate oligonucleotides bearing negative charges were adsorbed on positively charged emulsion composed of medium chain triglycerides. The physicochemical properties of the complexes were determined, as well as their stability in culture medium. Their interaction with erythrocytes through haemolysis, binding experiments and confocal microscopy were also evaluated. Finally, the in vitro evaluation of parasite growth and reinfection capacity was performed. The overall results showed that antisense oligonucleotides against P. falciparum topoisomerase II gene can be efficiently adsorbed onto a cationic nanoemulsion forming complexes. (Bruxel F, 2011)
· Melasma hyperpigmentation is an acquired disorder affecting the female population. The present study was to determine the ability of a botanical extract to reduce observable hyperpigmentation. The extract from heartwood of Artocarpus incisus was formulated into nanoemulsions, and the depigmenting efficacy of the formulated nanoemulsion was determined in vivo. The nanoemulsion containing the extract was formulated and prepared by the phase inversion technique. The depigmenting efficacy was observed following topical application of the formulated nanoemulsion to UVB-stimulated hyper pigmented dorsal skin of C57BL/6 mice. A strongly visible decrease in hyperpigmentation was observed after six weeks of treatment. The applied areas would return to their original colour after treatment was stopped for four weeks. (Buranajaree S, 2011)
· Nanoemulsion of amlodipine besilate was developed by spontaneous emulsification method to enhance the solubility and oral bioavailability of amlodipine besilate. The selected formulations from nanoemulsion region were subjected to droplet size analysis, partitioning study and in vitro drug release. The optimal formulation was characterized by transmission electron microscopy (TEM). The pharmacokinetics and bio- distribution studies of the optimized radiolabeled formulation in mice shows a relative bioavailability of 475% against amlodipine besilate suspension. In almost all the tested organs, the uptake of amlodipine besilate nanoemulsion was significantly higher than amlodipine besilate suspension, also confirming the efficacy of nano-sized formulation at therapeutic site. The residence time of nanoemulsion further signifies the advantage of nanoemulsions as drug carriers for enhancing bioavailability of amlodipine besilate.(Chhabra G, 2011)
· The aim of the current study was to develop a novel pea protein-based emulsifier which was used in the production of oil in water (O/W) nanoemulsions. The regions of kinetically stabilized O/W emulsions were individuated in a pseudoternary diagram containing different pea protein, sunflower oil, and water fractions. The stable O/W emulsion region was widened by the addition of NaCl to the aqueous phase or, to a more significant extent, by high pressure homogenization processing. The high pressure homogenization treatment caused the formation of very fine emulsions in the nanometric range (<200 nm), with extremely high stability over time. (Dosi F, 2010)
· Nanoemulsion based-1008 is a surfactant-stabilized soybean oil-in-water nanoemulsion adjuvant containing influenza antigen incorporated into the nanoemulsion by simple mixing. Intranasal administration of the antigen with nanoemulsion adjuvant efficiently produces both mucosal and serum antibody responses as well as a robust cellular Th1 immune response. To show the adjuvant effect of the W (80)5EC NE, a killed commercial influenza vaccine for intramuscular administration were mixed with the W (80)5EC NE adjuvant and administered intranasally to naïve ferrets. After a single intranasal immunization, the adjuvanted influenza vaccine elicited elevated serum hemagglutination inhibition (HAI) geometric mean titers (GMT) ranging between 196 and 905 for the three HA antigens present in the vaccine. This also produced significant cross immunity to five other H3N2 influenza strains not present in the vaccine, and produced sterile immunity. These findings shows the ability of W (80)5EC NE to adjuvant influenza vaccine when administered nasally provides the basis to study the intranasal W (80)5EC-adjuvanted influenza vaccine in humans. (Hamouda T, 2011)
· The aim of the study was to investigate the potential of food proteins as safer stabilizers for nanoemulsions to deliver hydrophobic drugs. Nanoemulsions stabilized by food proteins were prepared by high-pressure homogenization. The toxicity of the nanoemulsions was tested in Caco-2 cells using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazoliumbromide viability assay. In vivo absorption in rats was also evaluated. Food protein-stabilized nanoemulsions, with small particle size and good size distribution, exhibited better stability and biocompatibility compared with nanoemulsions stabilized by traditional emulsifiers. (He W,2011)
· The study aimed to develop a novel Amiloride loaded mucoadhesive nanoemulsion for nose-to-brain delivery. Furthermore, nasal irritation study and histopathological examination of the nasal mucosa were also carried out to assess non irritant nature of the nanoemulsion. Amiloride loaded mucoadhesive nanoemulsion formulations are non toxic on nasal mucosa and can be administered by intranasal route for effective treatment of epilepsy. (Jain N,2011)
· The aim of the current study was to test whether there is an increased efficacy of Dacarbazine as a nanoemulsion reducing tumour size in an epidermoid carcinoma xenograft mouse model. Tumours were induced in 5-week-old nude mice by subcutaneous injection. The mice were untreated or treated with a suspension of Dacarbazine (0.1 mg/kg), a nanoemulsion of Dacarbazine (0.1 mg/kg), every 2 days by either intramuscular injection (IM) or topical application. After 40 days, the final tumour size of mice receiving the nanoemulsion of Dacarbazine IM was significantly reduced compared to the suspension of Dacarbazine IM. The final tumour size of mice receiving the nanoemulsion of Dacarbazine topically was also significantly reduced compared to the suspension of Dacarbazine topically. This increased efficacy shows the greater stability of drug with the nanoemulsion in comparison with the suspension. (Kakumanu S, 2011)
· The present study was aimed to the evaluation, in vitro and in vivo, of glycyrrhetic acid release through the skin from the nanoemulsion system. Glycyrrhetic acid-loaded nanoemulsion was prepared by phase inversion temperature method, and was characterized in order to determine mean droplet size and its stability during a well-defined storage period. The Glycyrrhetic acid release pattern from nanoemulsion was evaluated in vitro; to determine its percutaneous absorption through excised human skin and in vivo evaluating Glycyrrhetic acid topical anti-inflammatory activity on healthy human volunteers by the UVB-induced erythema model. In vitro and in vivo evidence showed that the nanoemulsion system significantly increased the transdermal permeability of Glycyrrhetic acid in comparison to a control O/W emulsion containing the same amount of active compound. (Puliga G, 2010)
6.3
6.4
7.
7.1
/ OBJECTIVE OF THE STUDY
· The main objective of the study is to develop NSAID nanoemulsion.
· Evaluation of the formulated nanoemulsion.
· To increase the bioavailability.
PLAN OF WORK
· Survey of the literature.
· Procurement of chemicals.
· Formulation of the nanoemulsion.
· Evaluation of nanoemulsion.
· Stability studies of the formulated nanoemulsion.
· Thesis preparation.
MATERIALS & METHODS
SOURCE OF DATA
• Review of literature from:
Journals such as:
• International Journal of Pharmacy and Pharmaceutical Sciences.
• Journal of agriculture food chemicals.
• Journal of biomedical nanotechnology.
• Journal of cosmetic sciences.
• International Journal of Drug Delivery Technology.
• International Journal of Pharmaceutical Sciences and Nanotechnology.
Web sites :
• World Wide Web.
• Drug bank.com
• Science Direct
• Pub med
7.2
7.3
7.4
7.5
7.6
8
/ Materials
Drug and exciepients used will be procured from Pharma grade suitable manufacturer. Other reagents will be of Analytical grade.
Methods
· Spontaneous emulsification method is used for the preparation of this nanoemulsion.
· Emulsion phase inversion method.
· High pressure homogenization.
2) Evaluation of Nanoemulsions
a. Droplet size analysis.
b. drug release study.
c. Zeta potential.
d. Drug loading efficiency.
Method of collection of data (including sampling procedures if any)
The data will be collected from prepared formulations subjected to different evaluation techniques, scale-up techniques and stability studies obtained from ICH guidelines.
Does the study require any investigation or interventions to be Conducted on patients or other humans or animals?
- NOT APPLICABLE-
Has ethical clearance been obtained from your institution in case of 7.5?
NO
LIST OF REFERERENCES
1. Akhter S, Jalegaonkar S, Khan ZI, Jain GK, Khar RK, Ahmad FJ. Assessment of ocular pharmacokinetics and safety of Gancyclovir loaded nanoformulations. J Biomed Nanotech 2011 Feb, 7(1):144-5.
2. Alves PM, Pohlmann AR, Guterres SS. Semisolid topical formulation containing nimesulide loaded nanoemulsion: Development and rheological characterization. Pharmazie 2005 Dec; 60 (12) 900-4.
3. Amani A, York P, Chrystyn H, Clark BJ. Evaluation of a nanoemulsion based formulation for respiratory delivery of buclesonide by nebulizers. AAPS Pharm sci Tech. 2010 Sep; 11(3):1147-51.
4. Bruxel F, et al. Cationic nanoemulsion as a delivery system for oligonucleotides targeting malarial topoisomerase II. Int J pharm 2011 Feb 1.
5. Buranajaree S, Donsing P, Jeenapongsa R, Vivoch J. Depigmenting action of nanoemulsion containing heartwood extract of artocorpus incises on UVB-induced hyperpigmentation in C57BL/6 mice. J cosmetic sci 2011 Jan-Feb; 62(1)1-14.
6. Chhabra G, Chuttani K, Mishra AK, Pathak K. Design and development of nanoemulsion drug delivery system of amlodipine besilate for improvement of oral bioavailability. Drug dev Ind pharm Mar 2011 14.
7. Dosi F, Senatore B, Huang Q, Ferrari G. Development of a novel pea protein based nanoemulsions for delivery of neutraceuticals. J of Agric and Food chem. 2010 Oct 13; 58(19): 10653-60.
8. Hamouda T, Sutcliffe JA, Ciotti S, Baker JR jr. Intranasal immunization of ferrets with commercial trivalent influenza vaccines formulated in a nanoemulsion based adjuant. Clin Vaccine Immunol May 2011.
9. He w, Tan Y, Tian Z, Chen L, Hu F, Wu w. Food stabilized nanoemulsions as potential delivery systems for poorly water soluble drugs Preparation, in vitro evaluation and pharmacokinetics in rats. Int J Nanomedicine 2011 Mar 11 6:521-33.
10. Jain N, Akhter S, Jain GK, Khan ZI, Khar RK, Ahmad FJ. Antiepileptic intranasal Amiloride loaded mucoadhesive nanoemulsion: Development and safetyassessment. J Biomed Nanotechnol Feb 2011; 7(1):142-3.
11. Kakumanu S, Tagne JB, Wilson TA, Nicolosi RJ. A nanoemulsion formulation of Dacarbazine reduces tumour size in a xenograft mouse epidermoid carcinoma model compared to a Dacarbazine suspension. Nanomedicine Jan 2011 5.
12. Puliga C, Rizza L, Drechsler M, Bonina F. Nanoemulsions as vehicle for topical administration of glycyrrhetic acid characterization and in vitro and in vivo evaluation. Drug Deliv. Apr 2010 17 (3):123-9. .
9 / Signature of the Candidate / (ANJANIPRASADREDDY.VANGA)
10 / Remarks of the Guide / The topic selected for dissertation is satisfactory. Adequate equipments and chemicals are available to carry out the project work
11 / Name And Designation
11.1 / Guide / Dr. PRAKASH RAO.B
PROFESSOR &HOD
DEPARTMENT OF PHARMACEUTICAL TECHNOLOGY
KARNATAKA COLLEGE OF PHARMACY
#33/2, THIRUMENHALLI
HEGDE NAGAR MAIN ROAD
BENGALURU-64
11.2 / Signature of the Guide / (Dr. PRAKASHRAO.B)
11.3 / Co- Guide /
-NOT APPLICABLE-
11.4 / Signature of the Co- Guide / -NOT APPLICABLE-
11.5 / Head of the Department / Dr.PRAKASHRAO.B
PROFESSOR & HOD
DEPT OF PHARMACEUTICAL TECHNOLOGY
KARNATAKA COLLEGE OF PHARMACY
#33/2, THIRUMENHALLI
HEGDE NAGAR MAIN ROAD
BENGALURU-64.
S
11.6 / Signature of the HOD /
(Dr. PRAKASH RAO.B )
12 / 12.1 / Remarks of the Principal / All the required facilities will be provided to carry out dissertation work under the supervision of the Guide.
12.2 / Principal / Dr.K. RAMESH.
PRINCIPAL
KARNATAKA COLLEGE OF PHARMACY
#33/2, THIRUMENHALLI
HEGDE NAGAR MAIN ROAD
BENGALURU-64.
12.3 / Signature of the Principal / (Dr.K.RAMESH)
12