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RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA
ANNEXURE – II
PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION
1. NAME OF THE CANDIDATE
AND ADDRESS
Mr. SHAIKH MUZAMMIL SAGEER
DEPARTMENT OF PHARMACEUTICS,
K.L.E. SOCIETY’s COLLEGE OF PHARMACY,
J. N. M. C. CAMPUS, NEHRU NAGAR,
BELGAUM – 590010, KARNATAKA.
2. NAME OF THE
INSTITUTION
3. COURSE OF STUDY AND
SUBJECT
4. DATE OF ADMISSION TO
K.L.E. SOCIETY’s COLLEGE OF PHARMACY,
J. N. M. C. CAMPUS, NEHRU NAGAR,
BELGAUM – 590010, KARNATAKA.
MASTER OF PHARMACY IN
PHARMACEUTICS
JUNE 2007
COURSE
“STERICALLY STABILIZED
5. TITLE OF THE TOPIC
LIPOSOMAL DRUG DELIVERY
SYSTEM FOR DOXORUBICIN-DESIGN
AND CHARACTERIZATION”
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6. BRIEF RESUME OF THE INTENDED WORK:
6.1 NEED FOR THE STUDY:
The concept of drug delivery to a particular site for the treatment of localized disease
in the body and thereby decreasing adverse effect of drug with improving its therapeutic index
is being considered as a challenge in modern formulation design1. So, constant efforts have
been persued in order to design such an ideal drug delivery system which improves therapeutic
index of drugs and also improves patient compliance. One such area which has attracted
increasing attention of pharmaceutical scientist and has shown very promising results is
“Targeted Drug Delivery System”2.
Amongst all carriers utilized for target oriented drug delivery, vesicular drug delivery
system in form of liposomes is most extensively investigated. Liposome has been regarded as
a beneficial carrier system for parental application of variety of drugs due to their
biocompatible and biodegradable properties. Though liposomes have been proved to be
versatile carrier for wide variety of drug, there are two major inherent drawbacks of
conventional liposomes i.e.

Rapid uptake of drug carrier by phagocytic cells of RES.

Poor or shorter circulation time [3-5].
The inherent drawback of conventional liposome can be overcome by using a novel approach
of surface modification by using sterically stabilized liposomes or “Stealth Liposome”. The
most promising result of liposome modification were achieved for avoiding RES detection and
prolonging retention in circulation by covalent attachment of hydrophilic polymer groups to
the liposome surface to inhibit interaction with blood proteins and other plasma components
2
and thus liposome uptake by the macrophage system is inhibited. And there is an increase in
their circulation time. Longer circulation of liposome in blood resulted in higher uptake of
drug in an implanted tumor 6-8.
Doxorubicin is an anticancerous agent used in breast cancer, ovarian cancer, AIDS related
Kaposi’s sarcoma. The therapeutic efficacy of doxorubicin is restricted by dose limiting
toxicity to bone marrow and heart tissue9. The selective toxicity of doxorubicin could be
greatly improved if the concentration of drug in tumors could be increased relative to that in
sensitive normal tissues10.
Hence, in the present work, an attempt is being made to develop sterically stabilized liposomal
formulation of doxorubicin which will have following advantages.

Selective passive targeting to tumor tissues.

Increased efficacy and therapeutic index.

Reduction in toxicity of encapsulated drug.

Avoidance of RES uptake of drug carrier composite.

Increased circulatory time thus enhanced stability.
6.2 REVIEW OF LITERATURE
Literature review for undertaking the study was done by referring to various national
and international journals, published articles in various official standard books and referring to
various websites on the internet.
Attempt was done to study passive targeting of doxorubicin with polymer coated
liposomes in tumor bearing rats. The study revealed that polymers having a hydrophobic
moiety in the molecule such as PVA-R and HPMC-R are suitable materials for modifying the
surfaces of the doxorubicin-loaded liposome to improve its targeting property11.
Study on preparation, characterization and pharmacokinetics of N-palmitoyl chitosan
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anchored docetaxel liposome was done. The study revealed that anchored liposomes could
increase the stability of docetaxel in-vivo, as compared with plain liposomes, but the
improvement was not more significant than PEGylated liposomes. N-palmitoyl chitosan as a
new polymeric membrane to anchor liposome was useful to stabilize liposomes containing
anti-tumor drug12.
A study was conducted on Haloperidol-associated stealth liposomes. The study
demonstrated that haloperidol-targeted gene delivery systems can mediate efficient targeting of
genes to sigma receptor-over expressing breast cancer cells, thereby becoming a novel class of
therapeutics for the treatment of human cancers13.
An attempt was made to study Doxorubicin-loaded fab fragments of antidisialoganglioside immunoliposomes selectively inhibit the growth and dissemination of
human neuroblastoma in nude mice. It was concluded that fab-SIL[DXR] formulations led to
the total inhibition of metastatic growth of human NB in a nude mouse metastatic model. This
formulation should receive clinical evaluation as adjuvant therapy of NB14.
A study was conducted on therapeutic efficacies of isoniazid and rifampin
encapsulated in lung-specific stealth liposomes against Mycobacterium tuberculosis infection
induced in mice. The study stated that Liposome-encapsulated drugs at and below therapeutic
concentrations were more effective than free drug against tuberculosis, as evaluated on the
basis of CFUs detected, organomegaly, and histopathology. Furthermore, liposomal drugs had
marginal hepatotoxicities as determined from the levels of total bilirubin and hepatic enzymes
in serum. The elimination of mycobacteria from the liver and spleen was also higher with
liposomal drugs than with free drugs. The encapsulation of antitubercular drugs in lung
specific stealth liposome seems to be a promising therapeutic approach for the chemotherapy
of tuberculosis15.
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6.3 OBJECTIVES OF THE STUDY
1.
Preformulation studies.
2.
Preparation of Stealth liposomes by using Thin film hydration method or any
other suitable / appropriate method16.
3.
Characterization of formulated stealth liposomes for following parameters.
i.
ii.
Physical Characterization
a.
Vesicle size and surface morphology.----TEM
b.
Vesicle size and size distribution.---SEM17
c.
Surface charge.---ZETA MEATER 18
d.
Lamellarity.----NMR
e.
Phase behavior.----DSC 19
f.
In – Vitro drug release.
g.
In – Vivo tissue distribution study20.
h.
In – Vivo tissue tumor study
Chemical Characterizations.
a.
Phospholipids concentration by using HPLC.
b.
Cholesterol concentration by using HPLC.
c.
pH by using pH meter.
d.
Drug concentration by using appropriate methods as per the
individual monograph.
e.
Stability studies as per ICH guidelines21.
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7. MATERIALS AND METHODS
Drug
: Doxorubicin.
Carrier
: Egg-Phosphatidyl Choline (EPC), Dimyristoryl phosphatidyl choline
(DMPC), Distearoyl phosphatidyl choline (DSPC), PE-PEG.
Excipients
: Cholesterol, Stearyl amine, Polyvinyl Alcohol and its derivative
bearing a hydrophobic anchor (PVA-R)/ Hydroxy methyl cellulose
and its derivative having hydrophobic moiety (HPMC-R).
Method
: Preparation of Stealth liposomes by Thin film hydration method or
any other suitable / appropriate method16.
7.1 SOURCE OF DATA:
This data is obtained from the prepared Stealth liposomes drug delivery system of
doxorubicin based on the laboratory experiment and evaluation.
7.2 METHOD OF COLLECTION OF DATA:
i.
Physical Characterization
a.
Vesicle size and surface morphology----TEM.
b.
Vesicle size and size distribution----SEM17
c.
Surface charge----Zeta meter18
d.
Lamellarity.
e.
Phase behavior----DSC19
f.
In – Vitro drug release.
g.
In – Vivo tissue distribution study20.
h.
In – Vivo tissue tumor
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ii.
Chemical Characterizations.
a. Phospholipids concentration by using HPLC.
b. Cholesterol concentration by using HPLC.
c.
pH by using pH meter.
c. Concentration by using appropriate methods as per the individual
monograph.
e. Stability studies as per ICH guidelines21.
7.3
Does the study require any investigations or invention to be conducted
on patients or other human or animals? If so, please mention briefly.
Yes ( Wistar rats)
7.4 Has ethical clearance been obtained from your institution in
case of 7.3?
Yes
8. REFERENCES
1. Vyas SP, Singh RP, Jain S, Mishra V, Mohar S, Singh P et al. Non-ionic surfactant
based vesicles (niosomes) for non-invasive topical genetic immunization against
hepatitis-B. Int J Pharm 2005; 296:80-6.
2. Chein YW. Fundamentals of rate controlled drug delivery: Novel drug delivery system.
New York (NY): Marcel Dekker Inc.; 1992;43-137.
3. Poste G, Bucana C, Raz A, Bugelski P, Kirsh R, Fidler IJ. Analysis of the fate of
systemically administered liposomes and implications for their use in drug delivery.
Cancer Res 1982; 42:1412-22.
4. Senior JH. Critical reviews in therapeutic drug carrier systems. Biol Pharmaceut Bull
1987;3:123-93.
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5. Papahadjopoulos D., Gabizon A., Targeting of liposomes to tumor cells in vivo. Ann
NY Acad Sci 1987;507:64-74.
6. Poznansky M, Juliano RL. Biological approaches to the controlled delivery of drugs:
A critical review. Pharmacol Rev 1984;36:277-336.
7. Gabizon A, Papahadjopoulos D. Liposome formulations with prolonged circulation
time in blood and enhanced uptake by tumors. Proc Natl Acad Sci U S A 1988; 85(18):
6949-53.
8. Gabizon, A, Shiota R, Papahadjopoulos D. Pharmacokinetics and tissue distribution of
doxorubicin encapsulated in stable liposomes with long circulation times. J Natl
Cancer Inst 1989; 81(19):1484-8.
9. Allen TM, Mehra T, Hansen CB, Chin YC. Stealth liposomes: an improved sustained
release system for 1-β-D-arabinofuranosylcytosine. Can Res 1992; 52: 2431-2439.
10. Gabizon A, Catane R, Uziely B, Kaufman B, Safra T, Cohen R et al. Prolonged
circulation time and enhanced accumulation in malignant exudates of doxorubicin
encapsulated in polyethylene-glycol coated liposomes. Cancer Res 1994;54:987-92.
11. Takeuchi H, Kojima H, Yamamoto H, Kawashima Y. Passive targeting of Doxorubicin
with polymer coated liposomes in tumor bearing rats. Biol Pharm Bull 2001; 24(7):
795-9
12. Liang G, Jia-Bi Z, Fei X, Bin N. Preparation, characterization and pharmacokinetics of
N-palmitoyl chitosan anchored docetexal liposomes. J Pharm Pharmacol 2007; 59:
661-7.
13. Mukherjee A, Prasad TK, Rao NM, Banerjee R. Haloperidol-associated stealth
liposomes: a potent carrier for delivering genes to human cancer cells. J Biol Chem
2005; 280(16):15619-27.
14. Pastorino F, Brignole C, Marimpietri. Sapra P, Moase EH, Allen TM et al.
Doxorubicin-loaded fab, fragments of anti-disialonganglioside immunoliposomes
selectivity inhibit the growth and dissemination of human neuroblastoma in nude mice.
Cancer Res 2003;63:86-92.
15. Deol P, Khuller GK, Joshi K. Therapeutic efficacies of Isoniazid and Rifampin
encaposulated in lung-specific stealth liposomes against Mycobacterium tuberculosis
infection induced in mice. Antimicrob Agents Chemotherap 1997; 41(6):1211-4.
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16. Vyas SP. Khar RK. Targeted and Controlled Drug Delivery: Novel Carrier System.
New Delhi: CBS Publishers and Distributors; 2002:182-4.
17. Arunothayanun P, Unchebu IF, Craig DQM, Tutron JA. In vitro/in vivo
characterization of polyhedral niosomes. Int J Pharm 1999;183:57-61
18. Benoit JP, Bouligand Y, Lampercht A. “New lipid nanocapsules exhibit sustained
release properties for Amiodarone. J. Control Rel. 2002;84:59-68.
19. Molpeceres J, Aberturas MR, Guzman M. Biodegradable nanoparticles as a delivery
system for cyclosporine: preparation and characterization. J Microencap 2000;17(5):
599-614.
20. Prego C, Torres D, Megia FE, Caballal NR, Quinoa E, Alonso MJ. Chitosan –PEG
nanocapsules as new carriers for oral peptide delivery effect of chitosan pegylation
degree. J Control Rel 2006;111:299-308.
21. Carstensen JT. Drug stability: Principle and practice. New York: Marcel Dekker Inc;
1995:538-50.
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9.
Signature of Candidate
10.
Remarks of the guide
11.
The above information is true to the best of my knowledge and the work will be done under
my guidance.
Dr. RAJENDRA C. DOIJAD
11.1 Name and Designation of Guide
M. Pharm., Ph.D
PROFESSOR,
Department of Pharmaceutics,
K.L.E. Society’s College of Pharmacy,
J. N. M. C. Campus, Nehru Nagar,
Belgaum – 590010, Karnataka.
11.2 Signature
11.3 Co-Guide (IF ANY)
11.4 Signature
11.5 Head of the Department
Dr. F. V. MANVI M. Pharm., Ph.D
PRINCIPAL,
K.L.E. Society’s College of Pharmacy,
J. N. M. C. Campus, Nehru Nagar,
Belgaum – 590010, Karnataka.
11.6 Signature
12.
12.1 Remarks of the Chairman and Principal
The above mentioned information is correct and I recommend the same for approval.
12.2 Signature
Dr. F. V. MANVI M. Pharm., Ph.D
PRINCIPAL,
K.L.E. Society’s College of Pharmacy,
J. N. M. C. Campus, Nehru Nagar,
Belgaum – 590010, Karnataka.
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