FABRICATION AND CHARACTERIZATION OF LEVOFLOXACIN LUNGS TARGETED DRUG DELIVERY BY THE WAY OF MICROSPHERES
M. PHARM DISSERTATION PROTOCOL
SUBMITTED TO THE
RAJIV GANDHI UNIVERSITY OF HEALTH
SCIENCES, KARNATAKA. BENGALURU.
BY
ASHVINI H.M
B.Pharm.,
UNDER THE GUIDANCE OF
Mr. SHANKRAYYA. M
M.Pharm.,(Ph.D)
ASSISTANT PROFESSOR
P. G. DEPARTMENT OF PHARMACEUTICS
S. C. S. COLLEGE OF PHARMACY,
HARAPANAHALLI-583131
2011-12
Rajiv Gandhi University of Health Sciences, Bengaluru, Karnataka.
Annexure – II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
01 / Name and Address of the Candidate / ASHVINI H MD/O DWARAKASWAMY HIREMATH
# 43, IST WARD ,
GANESHA BADAVANE
HOSPET ROAD
HARAPANAHALLI-583131
DT;DAVANGERE. KARANATAKA
02 / Name of the Institution / T. M. A. E. Society’s
S. C. S. College of Pharmacy,
Harapanahalli – 583 131
(Davangere dist.) Karnataka
03 / Course of the Study
Branch / M. Pharm.
Pharamaceutics
04 / Date of Admission to course / 25/07/2011
05 / Title of the Topic / FABRICATION AND CHARACTERIZATION OF LEVOFLOXACIN LUNGS TARGETED DRUG DELIVERY BY THE WAY OF MICROSPHERES
06 /
Brief resume of the intended work
6.1. Need for the Study /Enclosure – I
6.2. Review of the Literature / Enclosure – II6.3. Objective of the Study
/ Enclosure – III07 /
Materials and Methods
7.1. Source of data /Enclosure – IV
7.2. Methods of collection of data
/ Enclosure – V7.3. Does the study require any
Investigations on animals?
If yes give details / Enclosure – VI
7.4. Has ethical clearance been
obtained from your institution
In case of 7.3. / Yes, Registration No: 157 / 1999 / CPCSEA
(Copies enclosed)
08 /
List of References
/ Enclosure – VII09 / Signature of the candidate / (ASHVINI H M.)
10 / Remarks of the Guide / Above mentioned title will be carried out in our college, in the Department of Pharmaceutics
11 / Name and Designation of
(In Block Letters)
11.1. Guide reference no. of RGUHS
ACA/CDC/PGT-
M.Ph/SCS/02/2010-11
11.2.Signature
11.3.Co-Guide (if any)
11.4.Signature
11.5. Head of the
Department
11.6.Signature / Mr. SHANKRAYYA. M
M. Pharm.,(Ph.D)
ASST. PROFESSOR
P.G. Dept. of Pharmaceutics,
S.C.S. College of Pharmacy,
Harapanahalli-583131,
Davanagere Dist.
Prof. VENKATESH. J.S.
B.Sc., M.Pharm.,( Ph. D)
Professor & Head
P.G. Dept. of Pharmaceutics,
S.C.S. College of Pharmacy,
Harapanahalli-583131.
Prof. VENKATESH. J.S.
B.Sc., M.Pharm.,( Ph. D)
Professor & Head
P.G. Dept. of Pharmaceutics,
S.C.S. College of Pharmacy,
Harapanahalli-583131.
12 / Remarks of the Principal
12.1. Signature / The candidate has been permitted to carry out the said above mentioned title of the work can be done into the department of the Pharmaceutics.
( Dr.R.Nagendra Rao)
ENCLOSURE-I
06. Brief resume of Intended Work
6.1 Need for the study.
Conventional oral drug administration does not usually provide rate-controlled release or target specificity. In many cases, conventional drug delivery provides sharp increase in drug concentration often achieving toxic level and following a relatively short period at the therapeutic level of the drug concentration eventually drops off until re-administration. In order to obtain maximum therapeutic efficacy, it becomes necessary to deliver an agent to the target tissue in the optimal amount for the required period of time, thereby causing little toxicity and minimal side effects. Desired drug release can be provided by rate controlling membranes or by implanted biodegradable polymers containing dispersed medication.
In pharmaceutical field, novel and controlled drug delivery system are becoming more popular which are capable of improving patient compliance as well as therapeutic efficacy. Controlled drug delivery refers specially to the precise control of the rate by which a particular drug dosage is released from a delivery system without the need for frequent administration.1,2
In recent years, much research in drug delivery has been focused on degradable polymer microspheres. Administration of medication via such systems is advantageous because microspheres can be ingested, injected or tailored for desired release profiles and in some cases it can provide organ-targeted release.
Microspheres are considered and accepted as a reliable one to deliver the drug to the target site with specificity, to maintain the desired concentration at the site of interest without untoward effects. Microencapsulation is a useful method which prolongs the duration of drug effect significantly and improves patient compliance. Eventually the total dose and few adverse reactions may be reduced since a steady plasma concentration is maintained.
Microspheres of biodegradable and non-biodegradable polymers have many advantages that they degrade in biological fluids. They can be injected, implanted and inserted into the body, they are non toxic and surgical removal of the polymer skeleton is not required. Therefore microspheres using various kinds of biodegradable polymers are designed to degrade as a result of hydrolysis of the polymer chains into biologically acceptable and progressively smaller compound. The success of any microencapsulation method depends on many factors such as the drug solubility, partition co-efficient, polymer composition, molecular weight etc3.
Majority of pharmaceutical formulations in contemporary use, their ‘specificity’ towards appropriate sites of a disease is not based on their ability to accumulate selectively in the target organ or tissue. Usually, they are more or less evenly distributed within the body. Moreover, to reach the target area, the drug has to
cross-different biological barrier organs, cells, even intracellular compartments, where it can cause undesirable side-reactions, or be partially inactivated. The best solution to this issue is drug targeting7.
Levofloxacin, a fluoroquinolone anti-infective, optically active L‐isomer of ofloxacin and two fold more potent than ofloxacin and reported to be more effective in the treatment of various bacterial infection.
Levofloxacin is available in the market as a conventional dosage forms such as tablets, capsules, and parenteral for the treatment of bacterial infections but not suitable means for the treatment of infection (pneumonia) locally4.
Hence, it is a challenge to develop injectable microspheres containing Levofloxacin for targeting to lungs and act locally on the organ of infection will improve the therapeutic efficacy, reduce side effects and thereby provide patient compliance.
ENCLOSURE-II
6.2 Review of Literature
Levofloxacin is BCS class I drug and dose is 500 mg/day having half life of 6-8 hours. So many successful attempts were carried out to convert Levofloxacin in sustain release dosage form. Microsphere is one of the greatest techniques for formulation of sustain release dosage form and many success stories are comply with it’s sustain release dosage form with various drugs in various dosage form. To give support of above discussion there are some literatures are cited below:
Ø Shen hong-liang, et al., prepared levofloxacin sustained-release microsphere with carboxymethyl chitosan by implementing emulsification and cross linking process. The prepared formulations were evaluated for the various parameters like particle size, surface characteristics, encapsulation efficiency and drug release pattern. The results of the drug release study shows that the appreciable sustained release of the drug5.
Ø Thakkar VT, et al., prepared levofloxacin hemihydrates floating tablets by direct compression method using Gelucire 43/01 (hydrophobic) and hydroxy propyl methyl cellulose (hydrophilic) polymer in different ratios. The floating tablets were evaluated for uniformity of weight, hardness, friability, drug content, in-vitro buoyancy and in vitro release studies. Various models were used to estimate kinetics of drug release. In-vitro release study reveals that the release rate of drug was decreased by increasing the proportion of Gelucire 43/01, 5 to 40%. The release rate of levofloxacin hemihydrates from matrices was mainly controlled by the hydrophilic and hydrophobic polymer ratio6.
Ø Sree Harsha et al., Prepared albumin loaded ofloxacin microspheres (ALOME) for lung-targeting, microspheres (ALOME) were prepared by water in oil emulsion method. The appearance and size distribution were examined by scanning electron microscopy, and the aspects such as in-vitro release characteristics, stability, drug loading, loading efficiency, pharmacokinetics and tissue distribution in albino mice were studied. The experimental results showed that the microspheres have an average particle size of 11.32µm. The drug loading and loading efficiency were(66.95 and 94.8%)respectively. The in-vitro release profile of the microspheres matched the Korsmeyer’s Peppas release pattern, and the release after 1 h was 42%, while for the original drug, ofloxacin, under the same conditions, 90.02% released in the first half an hour. After intravenous administration (15 min), the drug concentration of microspheres group in lung of albino mice was 432µgg−1 while that of controlled group was 1.32µgg−1 ALOME found to release the drug to a maximum extent in the target tissue, lung. Histopathological studies proved the stissue compatibility of ALOME to be safe7.
Ø Yalcin Ozakan, et al., prepared clarithromycin microspheres and evaluated by an emulision polymeration technique. Two matrix materials have been consider as the basis in preparing the selected model active substance. Natural human serum albumin(HAS) and bovin serum albumin(BSA), Albumin microspheres containing clarithromycin were prepared by heat stabilisation at different stirring rate. In the first part of our study, drug content, payload, particle size, surface morphology and release characteristic from microspheres prepared. The release of clarithromycin from both types of prepared microspheres was measured for 24 hr. The release profle of clarithromycin from HAS and BSA microspheres were extended for 24 hrs8.
Ø Muniyandy Saravanan, et al., made an attempt to target diclofenac sodium to its site of action through magnetic gelatin microspheres. The gelatin magnetic microspheres loaded with 8.9% w/w of diclofenac sodium and 28.7% w/w of magnetite were formulated by emulsification/cross-linking with glutaraldehyde. The formulated microspheres were characterized by particle size distribution, scanning electron microscopy (SEM), differential scanning calorimetry (DSC), X-ray diffraction and in vitro release studies. The in vivo distribution and targetability of gelatin magnetic microspheres after i.v. administration were studied in rabbits. The formulated microspheres were below 5 µm and spherical in nature as evidenced by the SEM photographs. DSC and X-ray diffraction studies revealed the absence of drug–polymer interaction. Encapsulated diclofenac sodium was released slowly more than 18 days9.
Ø Kadriye Ciftci, et al., prepared poly (DL-Lactic-Co-glycoside) PLAGA (50/50) microspheres containing an antineoplastic drug, 5- Fluorouracil (5-FU) by a solvent evaporation process in order to passively target liver carcinomas. The microspheres were spherical with diameter 2-5µm and encapsulated more than 70% (w/w) of the 5-FU. In-vitro release pattern of 5-FU from microspheres were determined for various systems. It was found that drug release depend upon the amount of entrapped drug, the polymer molecular weight and PH of the dissolution medium. The in-vitro release mechanism was diffusion controlled and followed a square-root of time relationship 10.
Ø Hiroaki Okada, prepared microspheres (msp) containing the LH-RH superagonist leuprorelin (leuprolide) acetate by using biodegradable polymers poly(lactic/glycolic acid) (PLGA) and poly(lactic acid) (PLA) as wall materials. A novel W/O/W emulsion-solvent evaporation method was devised for the preparation of msp containing this water-soluble peptide. This method achieved high entrapment efficiency and sustained drug release over a long period predominantly due to polymer bioerosion. The msp are fine microcapsules with polycores containing the peptide at a high concentration and are easily injectable through a conventional fine needle. Leuprorelin msp made with PLGA(75/25)-14,000 or PLA-15,000 released the drug in a zero-order fashion, maintained constant serum drug levels and attained persistent objective suppression of the pituitary-gonadal system (‘chemical castration’) over 1 or 3 months after i.m. or s.c. injection into animals. These resultsindicate that depot formulations may be potentially useful in the therapy of endocrine diseases in humans11.
Ø Sheth Zankhana, et al., developed biodegradable microparticulate delivery system of 5-fluorouracil by solvent evaporation technique by using polymethacrylate polymers like eudragit L100, eudragit S100, eudragit P4135F and methylcellulose. The formulations were evaluated with respect to particle size analysis, entrapment efficiency, in-vitro drug release studies, in-vivo drug targeting studies and stability studies. The formulated magnetic microspheres were found to be spherical with average particle size of 3-12 μm in diameter and incorporation efficiency up to 78.80%. In-vitro drug release
after 12 hr was 86.41 %, 92.84 %, 79.88 % and 82.38 % for formulation F1, F2, F3 and F4 respectively. Formulation F2 with highest drug content was selected for in-vivo drug targeting studies. The average targeting efficiency of drug loaded microspheres was found to be 26.16 % of the injected dose in liver, 11.40 % in lungs, and 15.08 % in spleen, whereas the concentration of pure drug was 15.52 % in liver, 9.0 % in lungs, and 9.50 % in spleen. These results reveal that the drug loaded microspheres showed preferential drug targeting to liver followed by spleen and lungs. Stability studies revealed that 4º C is the most suitable temperature for storage of 5 - fluorouracil loaded microspheres12.
Ø Mokhtar M. El-Basier, et al., studied in their investigation poly(L-lactic acid) (PLA) microspheres containing nedocromil sodium and beclometasone dipropionate (BDP), in-vitro lung model following aerosoliation from a dry powder inhaler. The in-virto kinetics of drug release releaved a controlled release of nedcromil sodium over 8 days with brust effect (27-60%,w/w) which varied with the particle size of microsphere. For BDP entrapped in PLA microspheres, controlled release of occurred over 6 days13.
Ø Jia Yu, et al., prepared biodegradable microspheres of the dexamethasone acetate with the polylactic-co-glycolic acid (PLGA). The microspheres were prepared using an oil-in-water emulsion technique. Physical properties, such as particle size, DMA entrapment efficiency, DMA release patterns and morphological characteristics were investigated by laser diffraction, HPLC, SEM method, and DSC in order to optimize the formulation of the microspheres. DMA was released immediately during the initial phase then, following this initial burst release, the microsphere system then released the drug in a continuous fashion. PLGA microspheres containing DMA is suitable for controlled release devices for use in the treatment of proliferative vitreoretinopathy (PVR) due to their size and drug release profile14.
Ø Meixia Jin, et al., prepared bovine serum albumin(BSA) alginate/ chitosan microspheres for the oral administration. Microspheres were prepared by a modified emulsifying-gelatinization method. The influence of the preparation conditions on the encapsulation efficiency, drug loading and yield of the microspheres was investigated by an orthogonal design method and the optimal process parameters were obtained. The in-vitro release of BSA from the alginate/chitosan microspheres was investigated in 0.1 M HCl solution (pH 1.2) and PBS (pH 7.4) as the release media15.