Synthesis and Biological Evaluation of

Pyrimidine Derivatives

M. Pharm Dissertation Protocol Submitted to

Rajiv Gandhi University of Health Sciences, Karnataka

Bangalore – 560 041

By

Mr. ABHIK DEBBARMAN

Under the Guidance of

PROF. D. GILES

Department of Pharmaceutical Chemistry,

Acharya & B.M. Reddy College of Pharmacy,

Soldevanahalli, Chikkabanavara Post,

Bangalore -560090.

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

KARNATAKA, BANGALORE.

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1 /

Name of the candidate and address

/

Mr. ABHIK DEBBARMAN

House No.609, 7th Cross,
Near Union Bank, AGB Layout.
Hesarghatta main road,
Bangalore-560090.
2 / Name of the institution / ACHARYA & B.M. REDDY COLLEGE OF PHARMACY.
89/90, Soldevanahalli, Chikkabanavara post, Hesaraghatta main road,
Bangalore – 560090.
Office: 080-28396011
3 / Course of study and subject /

MASTER OF PHARMACY

(PHARMACEUTICAL CHEMISTRY)
4 / Date of the admission / 10th JANUARY 2011
5 /

Title of the topic:

“SYNTHESIS AND BIOLOGICAL EVALUATIONOF PYRIMIDINE DERIVATIVES.”
6.0 / Brief resume of the intended work
Need for study:

Pyrimidine is the most important member of all the diazines as the ring system occurs widely in living organisms.It exhibit a broad spectrum of biological activities such as analgesic, 1-9anti inflammatory, 10-12antimicrobial, 11-17anticancer activities18-34etc.
Prostaglandins, arachidonic acid metabolites of cyclooxygenase pathway, are major mediators in regulation of inflammation and immune function. Cyclooxygenase (COX) is well known to be responsible for prostaglandins production and the rate-limiting enzyme. This enzyme exists in two isoforms, namely COX-1 and COX-2 and induces inflammation cytokines, including tumor necrosis factor-α, interleukin and lipopolysaccharide10. Because COX-2 has a crucial role in the carcinogenesis and progression of cancer exploration of its mechanisms can lead to new therapeutic targets to improve survival of cancer patients. Celecoxib, a selective COX-2 inhibitor has been shown to possess the highest anticarcinogenic capacity of nonsteroidal anti-inflammatory activity12
Kinase is a type of enzyme that transfer phosphate groups from high energy donor molecules, such as ATP to specific substrate, a process referred to as phosphorylation. Protein kinases are targets for treatment of a number of diseases. Kinase inhibitors are useful in the treatment of cancer and inflammation. Some of the kinase inhibitors were used in the treatment of cancer18-31.
Marketed Drugs Containing Pyrimidine Nucleus:-
Anti-inflammatory & Analgesic agents


Marketed Drugs Containing Pyrimidine Nucleus:-Anticancer Agents:-



Several pyrimidine derivatives are found to be COX-2 and kinase inhibitors. By considering the above facts it was planned to synthesize pyrimidine derivatives and evaluated for its analgesic, anti-inflammatory and anticancer activity.
Objectives of the study
1)To synthesize some derivatives of pyrimidine.
2)To characterize the synthesized compounds by different analytical techniques such as IR, 1H NMR and mass spectral data.
3)To screen the synthesized compounds for their in vivo anti-inflammatory,in vivo analgesic andin vivo anticanceractivities.
4)To publish the research works in peer reviewed journals.
ACTIVITIES / DURATION
Literature survey / Till the completion of project
Synthesis and collection of analytical data / 6 Months
Pharmacological activities / 2 Months
Typing of thesis book and sending for publication / 1 Month
Total / 9 Months
7.0
8.0 / Materials and methods:
Sources of data
Databases like Chemical abstracts, Biological abstracts, Medline, and Journal of Chemistry section B, Indian Journal of Heterocyclic Chemistry, European Journal of Medicinal Chemistry, Bioorganic and Medicinal Chemistry Letters, Acta crystallographica, through Helinet of RUGHS etc.
Method of collection of Data
A) Synthesis of the compounds:
Chemicals and other reagents required for the synthesis will be procured from standard company sources. Compounds will be synthesized by using standard procedures. The reactions will be monitored by TLC and purification of the compounds will be carried out by recrystallization method using suitable solvent.
SCHEME FOR SYNTHESIS:36
Step I:

Step II:

B) Characterization of the compounds:
The synthesized compounds will be characterized by preliminary laboratory techniques such as melting point, boiling point etc and by FTIR,1H NMR and mass spectroscopy spectral data.
C) Biological Evaluation:
1)Acute toxicity studies: To determineLD50 of the test compound.
2) Screening of anti-inflammatory activity:37-39
Invivo anti-inflammatory activity study:
Method Used: Carrageenan-induced paw edema model.
Animals Used: Albino Wistar rats.
Carrageenan-induced paw edema model:
A 1% w/v suspension of carrageenan will be prepared freshly in normal saline and injected into subplantar region of left hind paw (usually 0.1mL in rats and 0.025-0.05mL in mice). In control group animals, only vehicle will be injected. Test drug is usually administered orally or intraperitoneally, according to body weight immediately or half an hour or one hour before (depending on the expected peak effect) carrageenan challenge. A mark will be made on the ankle joint of each rodent. Paw volume up to the ankle joint will be measured in drug treated and untreated groups before and after 3 h of carrageenan challenge using a plethysmograph filled with mercury.
3) Screening of analgesic activity:40
Invivo analgesic activity study:
Method Used: Acetic acid Writhing method.
Animals Used: Swiss Albino mice.
Acetic-acid induced writhing in mice:
The animals will be weighed, numbered and divided into groups of five animals in each group. Appropriate volume of acetic acid solution willbe administer to the first group (which serves as control) and placed individually under glass jar for observation. The onset on writhes willbe noted. The number of abdominal contractions, trunk twist response and extension of hind limbs as well as the number of animals showing such response during a period of 10 min is recorded. The test compound will be injected to the second group of animals. Fifteen minutes later, acetic acid solution will be administered to these animals. The onset and severity of writhing response will be noted. The mean writhing scores in control and compound treated groups are calculated and inhibition of pain response by compound will be noted.
4)Screening of anticancer activity:41-43
Anticancer activity study: Against Ehrlich Ascites Carcinoma(EAC cells)
Method Used: In-vivoanticancer activity
Animals Used: Swiss Albino mice
Anticancer activity against Ehrlich Ascites Carcinoma:
The animals will be divided into different groups of 6 animals each depending on the test compound synthesized . The EAC cell containing phosphate buffer saline (106 cells/0.1 mL) will be injected into the peritoneal cavity of test group animals and treatment will be started 24 h after inoculation of tumor cells, (once daily as single dose) for 10 days.
Group I will serve as control and will receive 0.3% CMC suspension. Group II will serve as standard and will receive vincristine (ip, 520 µg/kg body weight). Remaining groups will serve as test groups and receive test compound, administered intraperitoneally. Antitumor activity will be screened by determining different parameters like body weight analysis, mean survival time and percent increase in life span and also haemotological paramaters like Hb, RBC, WBC, Lymphocytes and Monocytes.
Group II will serve as standard 5-Flurouracil (520 μg/kg body weight). Group III-XII serve as test groups and treated with test compounds. The control, standard and test compounds will be administered intraperitoneally. Antitumor activity will be screened by determining different parameters like body weight analysis, mean survival time and percentage increase in life span.
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