Download “Formulation, Development and Evaluation of Controlled and

“Formulation, Development and Evaluation of Controlled and
Targeted release of Tamoxifen in the Treatment of Breast
Cancer.”
DISSERTATION PROTOCOL
SUBMITTED TO
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BANGALORE, KARNATAKA.
BY
Shravan L. Nargund
M.PHARM, PART-I
DEPARTMENT OF PHARMACEUTICS
NARGUND COLLEGE OF PHARMACY
BANGALORE-85, KARNATAKA
(2011-2013)
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1.
NAME OF THE CANDIDATE
AND ADDRESS (IN BLOCK
LETTERS)
2.
NAME OF THE INSTITUTION
3.
COURSE OF STUDY AND
SUBJECT
4.
DATE OF ADMISSION OF
COURSE
5.
TITLE OF TOPIC
Shravan L. Nargund
#2366 ,12th MAIN ,A BLOCK, 2nd STAGE ,
RAJAJINAGAR
BANGALORE -560010
NARGUND COLLEGE OF PHARMACY,
DATTATREYANAGAR, II MAIN,
100 FT RING ROAD, BSK III STAGE,
BANGLORE-85
KARNATAKA.
MASTER OF PHARMACY IN
PHARMACEUTICS
27th JULY 2011
“FORMULATION, DEVELOPMENT
AND EVALUATION OF
CONTROLLED AND TARGETTED
RELEASE OF TAMOXIFEN IN THE
TREATMENT OF BREAST CANCER.”
6. BRIEF RESUME OF THE INTENDED WORK:
6.1 NEED FOR THE STUDY:
Cancer is one of the deadliest disease the modern world has experienced, accounting for
nearly 7.6 million deaths (around 13% of all deaths) in the year 2008, worldwide. Lung, stomach,
liver, colon and breast cancer account for the most cancer deaths every year. About 30% of cancer
deaths are due to five leading behavioral and dietary risks, high body mass index, low fruit and
vegetable intake, lack of physical activity, tobacco use, alcohol use with about 70% of all cancer
deaths occurring in low and middle income countries. Deaths from cancer worldwide are
projected to rise to over 11 million in 2030.
Breast cancer is one of the leading causes of deaths in women. Breast cancer caused
458,503 deaths worldwide (13% of cancer death in women) and comprises 22.9% of all types of
cancers. The first recorded cancer in the world dates back to 1500 BC from ancient Egypt,
wherein the tumors in the breast were treated by destroying the tissue with hot instruments. Even
after so many years direct cure for this form of cancer is limited. Breast cancer nowadays is
treated by surgery, by medication, chemotherapy and by using monoclonal antibodies.
Medication basically involves using different drugs like tamoxifen, anastrozole, letrozole to
inhibit the growth of cells by acting on the estrogen receptors which releases the growth
hormones. Tamoxifen is the most widely used drug which has a dosage regime of 20 mg per day
for 5 years before surgery, nowadays it is also used after surgery.
Tamoxifen’s primary metabolite trans-4-hydroxytamoxifen is equally active as the parent
drug; hence this is taken into consideration to reduce the dosage regime of the drug. The activity
of the drug can be enhanced by targeting the drug to the specific site of action and further
controlling the release rate of the drug. Although targeting the drug to the particular tissue is a
challenge, it is envisaged that by evaluation of suitable polymers and/or combinations thereof will
help to overcome the challenge.
Thus the need of the study is formulating an effective product which will lead to increase the
patient compliances, reduce dosage regime and decreasing side effects by achieving all the above
tasks.
6.2 REVIEW OF LITERATURE :
Literature shows that limited work has been reported on targeted and controlled release
of tamoxifen. Some of them are listed below
Jugminder S Chawla, Mansoor M Amji (2002) increased the concentration of tamoxifen in
estrogen receptor in positive breast cancer by developing nanoparticle formulation using poly
(ᵋcaprolactone)(PCL). The nanoparticles were prepared by solvent displacement method using
acetone-water system. Particle size analysis, scanning electron microscopy, zeta potential
measurements and differential scanning calorimetry were used for nanoparticle characterization.
Biodegradation studies were performed in the presence and absence of Pseudomonas lipase in
phosphate buffered saline (PBS , pH 7.4) at 37 degrees C. Tamoxifen loading over different
concentrations was analyzed by high performance liquid chromatography (HPLC) and the
optimum loading concentration was determined.
Jayanta
K. Sarmah,
Rita Mahanta,
Saibal
Kanti Bhattacharjee,
Ranadeep Mahanta, Angshuman Biswas (2011) reported the preparation of Controlled release
tamoxifen citrate encapsulated in cross-linked guar gum nanoparticles. They used Natural
polysaccharides, which due to their merits, have received more and more attention in the field of
drug delivery. In the present study tamoxifen citrate, TMX (a non-steroidal antiestrogenic drug)
loaded guar gum nanoparticles, GG NPs, cross linked with glutaraldehyde were prepared for
treatment of breast cancer. Oil in water (o/w) emulsion polymer cross-linking method was
employed for preparation of blank and drug loaded sustained release nature biodegradable
nanoparticles. Prepared nanoparticles were characterized by morphology in scanning electron
microscope (SEM), size distribution in transmission electron microscope (TEM), TMX loading by
high performance liquid chromatography (HPLC) and in vitro drug release characteristics. An
overall sustained release of the drug from the biodegradable nanoparticles was observed in in vitro
release studies.
A group of German scientists, Wilfried Fischer, Karin Klokkers, Anna Sendl-Lang (1999)
patented transdermal system in the form of a patch comprising of tamoxifen derivative.. As
tamoxifen derivatives like 3-hydroxy tamoxifen or 4-hydroxy tamoxifen are very lipophylic
suitable additives are added to increase its penetration through stratum corneum. They are usually
treated with alcohol to make it in alcoholic or aqueous solution..
Gordon Nelson Arbuthnot, Karen Klapper Lomas, and Glenn Alan Meyer (2001), prepared a
pharmaceutical formulation, which comprised 5-15% by weight of tamoxifen citrate and a 1st
disintegrant which is croscarmellose sodium. The percentage by weight of croscarmellose sodium
present in the formulation was chosen such that in six 20 mg unit doses, an average of at least
65% by weight of the tamoxifen citrate in the formulation will be dissolved within 10 mins. in
1000ml of 0.02N HCl acid at 37degrees C when stirred at 100 rpm.
Avnesh Kumari, Sudesh Kumar Yadav, Subhash C. Yadav (2009) have published their work
on biodegradable polymeric nanoparticles based drug delivery systems.. Biodegradable
nanoparticles have been used frequently as drug delivery vehicles due to its greater
bioavailability, better encapsulation, control release and less toxic properties. Various
nanoparticulate systems, general synthesis and encapsulation process, control release and
improvement of therapeutic value of nanoencapsulated drugs are covered in this review. They
have highlighted the impact of nanoencapsulation of various disease related drugs on
biodegradable nanoparticles such as PLGA, PLA, chitosan, gelatin, polycaprolactone and polyalkyl-cyanoacrylates.
Dinesh B. Shenoy and Mansoor M. Amiji (2005) have evaluate and compared the
biodistribution profile of tamoxifen when administered intravenously (i.v.) as a simple solution or
when encapsulated in polymeric nanoparticulate formulations, with or without surface-stabilizing
agents. Tamoxifen-loaded, poly(ethylene oxide)-modified poly(ɛ-caprolactone) (PEO-PCL)
nanoparticles were prepared by solvent displacement process that allowed in situ surface
modification via physical
adsorption
of
poly(ethylene oxide)–poly(propylene oxide)–
poly(ethylene oxide) (PEO–PPO–PEO) triblock polymeric stabilizer (Pluronic®). The
nanoparticles were characterized for particle size and surface charge. Presence of PEO chains on
nanoparticle surface was ascertained by electron spectroscopy for chemical analysis (ESCA). In
vivo biodistribution studies were carried out in Nu/Nu athymic mice bearing a human breast
carcinoma xenograft, MDA-MB-231 using tritiated [3H]-tamoxifen as radio-marker for
quantification. PEO-PCL nanoparticles with an average diameter of 150–250 nm, having a
smooth spherical shape, and a positive surface charge were obtained with the formulation
procedure. About 90% drug encapsulation efficiency was achieved when tamoxifen was loaded at
10% by weight of the polymer.
Otilia M. Koo, Israel Rubinstein, and Hayat Onyuksel (2006) have reviewed the role of
nanotech in the targeted drug delivery and imaging. The emphases of this review are on
biophysical attributes of the drug delivery and imaging platforms as well as the biological aspects
that enable targeting of these platforms to injured and diseased tissues and cells. The principles of
passive and active targeting of nanosized carriers to inflamed and cancerous tissues with increased
vascular leakiness, overexpression of specific epitopes, and cellular uptake of these nanoscale
systems are discussed. Preparation methods—properties of nanoscale systems including
liposomes, micelles, emulsions, nanoparticulates, and dendrimer nanocomposites, and clinical
indications are outlined separately for drug delivery and imaging in vivo. Taken together, these
relatively new and exciting data indicate that the future of nanomedicine is very promising.
8 Harivardhan Reddy (2010) studied recent statergies of drug delivery to tumours. Despite
several advancements in chemotherapy, the real therapy of cancer still remains a challenge. The
development of new anti-cancer drugs for the treatment of cancer has not kept pace with the
progress in cancer therapy, because of the non-specific drug distribution resulting in low tumour
concentrations and systemic toxicity. The main hindrance for the distribution of anti-cancer agents
to the tumour site is the highly disorganized tumour vasculature, high blood viscosity in the
tumour, and high interstitial pressure within the tumour tissue. Recently, several approaches such
as drug modifications and development of new carrier systems for anti-cancer agents have been
attempted to enhance their tumour reach. Approaches such as drug delivery through enhanced
permeability and retention (EPR) effect have resulted in a significant improvement in
concentration in tumours, while approaches such as drug-carrier implants and microparticles have
resulted in improvement in local chemotherapy of cancer. This review further discusses different
strategies employed for the delivery of anti-cancer agents to tumours, such as through EPR effect,
local chemotherapeutic approaches using drug delivery systems, and special strategies such as
receptor-mediated delivery, pH-based carriers, application of ultrasound and delivery to resistant
tumour cells and brain using nanoparticles.
Ikeda K. ,Inone (2004) studied estrogen receptor and their down stream targets in cancer.
Estrogen has crucial roles in the proliferation of cancer cells in reproductive organs such as the
breast and uterus. Estrogen-stimulated growth requires two estrogen receptors (ERalpha and
ERbeta) which are ligand-dependent transcription factors. High expression of ERs is observed in a
large population of breast tumours. In addition, the positive expression of ERs correlates with
well-differentiated tumours, a favourable prognosis, and responsiveness to an endocrine therapy
with anti-estrogen drugs in patients with breast cancer. Transcription activities of ERs can be
regulated by interacting proteins such as coactivators and kinases as well as ligand-binding.
Moreover, ER isoforms lacking an ability to transactivate are involved in breast cancer.
Downstream target genes of ERs have important roles in mediating the estrogen action in breast
cancer. We have isolated and characterized several novel estrogen-responsive genes to clarify the
molecular mechanism of the estrogen action in target cells. Among these genes, the estrogenresponsive finger protein (Efp) was found to be highly expressed in breast cancer. Efp as a
ubiquitin ligase (E3) is involved in the proteasome-dependent degradation of the 14-3-3sigma
protein, one of cell cycle brakes, this degradation resulting in the promotion of breast cancer
growth. A full understanding of the expression and function of ERs and their target genes could
shed light on how estrogen stimulates the initiation and promotion of cancer, providing a new
approach to diagnose and treat cancer.
Cheul-seong Kim, Sang-Jun Choi , Chi- Young Park et.al. (2010) studied the effects of
silybinin on the pharmacokinetic of tamoxifen and its active metabolite 4-hydroxytamoxifen in
rats. The effects of silybinin, an antioxidant, on the pharmacokinetics of tamoxifen and its
metabolite, 4-hydroxytamoxifen, were investigated in rats. A single dose of tamoxifen was
administered intravenously (2 mg/kg) and orally (10 mg/kg) without or with silybinin (0.5, 2.5
and 10 mg/kg) to rats. Silybinin significantly altered the pharmacokinetics of orally administered
tamoxifen. Compared to those in the oral control group (given tamoxifen alone), the area under
the plasma concentration-time curve (AUC0-∞) and the peak plasma concentration (Cmax) of
tamoxifen were significantly (p<0.05 for 2.5 mg/kg, p<0.01 for 10 mg/kg) increased by 40.271.3% and 45.2-78.6%, respectively, with silybinin. Consequently, the absolute bioavailability
(AB) of tamoxifen in the presence of silybinin (2.5 and 10 mg/kg) was 31.1-38.1%, which was
significantly enhanced (p<0.05) compared to that in the oral control group (22.2%). Moreover, the
relative bioavailability (RB) of tamoxifen was 1.40- to 1.72-fold greater than that in the control
group. Silybinin (10 mg/kg) significantly increased the AUC0-∞ (p<0.05, 40.0%) of 4hydroxytamoxifen, but the metabolite-parent ratio (MR) of 4-hydroxytamoxifen was significantly
altered (p<0.05 for 10 mg/kg), implying that the formation of 4-hydroxytamoxifen was
considerably affected by silybinin.
6.3 OBJECTIVES OF THE STUDY:
The present study is aimed to design and evaluate controlled and targeted release of tamoxifen.
The main objectives of the above study are:-
-To develop a suitable formulation for the controlled release of the drug.
-To develop a suitable method to target the drug to the specific site where the action is required.
- To attempt and reduce the dosage of the drug to be administered by meeting the above
requirements.
-To evaluate all the above with the help of suitable in vitro evaluation studies.
MATERIALS AND METHODS:
7.
Materials:
Polymers: Different polymers like PHEA-C16, PEG’s, Trimethyl chitosan, PCL (poly
caprolactone, PEO-PCL etc.
Drug: Antineoplastic drug used in the treatment of breast cancer, Tamoxifen will be used to
achieve the objectives of the study.
Excipients: The excipients may include Lactose monohydrate, magnesium state, Povidone,
Estradiol, SERM etc.
Methods:
The following methods can be used for the formulation of core in cup tablet.
Step-1 Formulation of the tablet.
Step-2 Coating with the polymers.
Step-3 Preparation of nanoparticles.
Step-4 Compression of tablets after addition of targeting substances.
7.1. Source of Data
 Library: Nargund College of pharmacy.
 E-library: Nargund College of pharmacy.
 Official books (IP, BP, USP and EP).
 RGUHS Library, Bangalore.
 International and national Pharmaceutical abstracts etc.

Journals & articles :
o
American chemical society.
o
International Journal of Pharmaceutics.
o
Journal of Controlled Release.
o
European Journal of Pharmaceutics.
o
Asian Journal of Pharmaceutical and Clinical Research
 Internet:.
 www.nanocancer.com
 www.google.com
 www.breast-cancer-research.com
 www.drugbank.com
 www.wikipedia.com
C. Text Books
o Raymond C Rowe, Paul J Sheskey and Sian C Owen. Hand book of pharmaceutical
excipients; Fifth edition.
o Herbert A. Lieberman, Leon Lachman and Joseph B.Schwatz. Pharmaceutical dosage
forms: Tablets volumes 1, 2 & 3.
o
Leon Lachman, Herbert A. Lieberman, Joseph L. Kanig. The theory and practice of
Industrial Pharmacy: 3rd edition.
o Goodman & Gilman’s “The pharmacological basis of theraputics” 10th edition,
international edition.
o Tripathi KD. Essential of medical pharmacology. 5th ed. New Delhi. Jypee brother; 2004.
o
Yie W. Chien “Novel drug delivery systems” 2nd edition revised and expanded.
7.2 Method of Collection of Data
 Literature survey using internet and scientific journals.
 Experimental studies which includes:
 The formulation evaluated for the compatibility studies using FTIR etc.
 Evaluating the pre-formulation parameters like bulk density, tapped density,
Carr`s index etc.
 Formulation development and evaluation.
 Granular properties, tablet properties and in-vitro dissolution studies for the
Developed formulation.
 UV/Vis Spectrophotometeric, HPLC etc. for the estimation of drug in in-vitro
Dissolution samples.
.
7.3. Does the study require any investigation or intervention to be conducted on
patients or other humans or animals? If so, please mention briefly.
-NO-
7.4. Has ethical clearance been obtained from your institution in case of 7.3?
- NOT APPLICABLE-
8. LIST OF REFERENCES:
1. Globocan 2008 , IARC 2010 Statistics.
2. who.int/media centre/factsheets/fs297/en/
3. cancer.about.com/od/historyofcancer/a/cancerhistory.htm.
4. Hariharan M, Gypta VK. A novel compression-coated tablet dosage form. Pharmaceutical
technology yearbook. 2001:14-9.
5.Ikeda K., Inone S. “Estrogen receptors and their downstream targets in the cancer”,
Archives of Histology and Cytology.[2004,67(5);435-42]
6. US patents by Gordon Nelson Arbuthnot, Indianapolis, IN(US) Karen Klapper Lomas,
Glenn Alan Meyer, both of Wilmington NC (US) patent number US 6,245,352 B1 on
Pharmaceutical Formulation of tamoxifen.
7. US patents by Wilfried Fischer, Karin Klokkers, and Anna Sendl-Lang all of
Holzkrichen Germany patent number 5,904,930 on transdermal systems in the form of a
patch comprising tamoxifen derivatives.
8. Jugminder S. Chawla, Mansoor M Amiji “biodegradable poly (ᵋ-caprolactone)
nanoparticles for tumour targeted delivery of tamoxifen.” International journal of
pharmaceutics.[volume 249,Issues1-2, 5 december 2002, pages 127-138].
9. Avnesh Kumari, Sudesh Kumar Yadav, Subhash C. Yadav. “Biodegradable polymeric
nanoparticles based drug delivery systems” Colloids and surfacesB:Biointerfaces[volume
75, issue1,1st January 2010, pages 1-18]
10. Otilia M. Koo, Israel Rubinstein, Hayat Onyuksel. “Role of nanotechnology in
targeted drug delivery and imaging: a concise review” Nanomedicine: Nanotechnology,
Biology and Medicine.[volume 1, Issue 3, September 2005, pages 193-212].
11. L. Harivardhan Reddy “Drug delivery to tumours ; recent strategies” Journal of
pharmacy and pharmacology.[volume57,Issue10, Pages 1231-1242,October 2005]
12. Jayanta K. Sarmah, Rita Mahanta, Saibal Kanti Bhattacharjee et.al. “Controlled release
of tamoxifen citrate encapsulated in cross linked guar gum nanoparticles” International
Journal of pharmaceutics.
13. Cheul-Seong Kim, Sang Jun Choi, Chi-Young Park et.al.”Effects of silybinin on the
pharmacokinetic of tamoxifen and its active metabolite 4-thydroxytamoxifen in rats”
Anticancer Research [January 2010, volume 30,no.1 79-85.
9.
Signature of the candidate
(SHRAVAN L. NARGUND)
10.
Remarks of the Guide
11.
Name & Designation of
(in block letters)
11.1 Guide
RECOMMENDED FOR THE DISSERTATION
WORK.
Dr. GOPAL MURALIDARAN
PROFESSOR
DEPARTMENT OF PHARMACEUTICS.
NARGUND COLLEGE OF PHARMACY.
11.2 Signature
11.3 Head of the department
DR. C.S.R. LAKSHMI
PROFESSOR
DEPARTMENT OF PHARMACEUTICS
NARGUND COLLEGE OF PHARMACY
11.4 Signature
C.S.R. LAKSHMI
12.
12.1 Remarks of Principal
FORWARDED AND RECOMMENDED FOR
FAVOURABLE CONSIDERATION.
12.2 Signature.
DAYANAND.S.PURANIK
(Principal)