“DEVELOPMENT OF ANALYTICAL METHOD FOR

QUANTITATIVE ESTIMATION OFSITAGLIPTIN

IN BULK AND PHARMACEUTICAL FORMULATIONS”

SYNOPSIS FOR

M.PHARM DISSERTATION

SUBMITTED TO

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES

KARNATAKA

BY

PATIL SACHIN LALASAHEB

B. Pharm

UNDER THE GUIDANCE OF

Dr B.RAMESH

M.Pharm Ph.D

DEPARTMENT OF PHARMACEUTICAL CHEMISTRY

S.A.C.COLLEGE OF PHARMACY

B.G.NAGARA, KARNATAKA -571448

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERATION

1 / Name of the candidate & Address / PATIL SACHIN LALASAHEB
S.A.C.COLLEGE OF PHARMACY
B.G.NAGARA-571448
KARNATAKA
PERMANENT ADDRESS:
SACHIN L PATIL
A/P: GHONASHI
KARAD (TALUKA)
SATARA (DIST)
STATE-MAHARASHTRA
2 / Name of the institute / S.A.C.COLLEGE OF PHARMACY
B.G.NAGARA
KARNATAKA -571448
3 / Course of the study / MASTER OF PHARMACY (Pharmaceutical chemistry)
4 / Date of admission / 15/06/2008
5 / Title of the topic / DEVELOPMENT OF ANALYTICAL METHOD FOR QUANTITATIVE ESTIMATIONOF
SITAGLIPTIN IN BULKAND
PHARMACEUTICAL FORMULATIONS.
6 / BRIEF RESUME OF THE INTENDED WORK
GENERAL DISCUSSION:
Sitagliptin phosphate is an oral antihyperglycemic (anti-diabetic drug) of the dipeptidyl peptidase-4 (DPP-4) inhibitor class. This enzyme-inhibiting drug is used either alone or in combination with other oral antihyperglycemic agents (such as metformin or thiazolidinedione) for treatment of diabetes mellitus type 2. The benefit of this drug is its lower side-effects (e.g., less hypoglycemia, less weight gain) in the control of blood glucose values.
STRUCTURAL FORMULA:

NOMENCLATURE: (3R)-3-amino-1-[9-(trifluoromethyl)-
1,4,7,8-tetrazabicyclo[4.3.0]nona-6,8-d ien-4-yl]-
4-(2,4,5-trifluorophenyl)butan-1-one
MOLECULAR FORMULA:C16H15F6N5O
MOLECULAR MASS: 407.314 g/mol
DESCRIPTION: White or off white crystalline powder.
SOLUBILITY: It is soluble in water and N, N dimethyl formamide, slightly soluble in methanol, soluble in ethanol, acetone and acetonitrile and insoluble in isopropanol and isopropyl acetate.
ANALYTICAL FUCTIONAL GROUPS: Amino, Trifluorophenyl .
Triazolopiperazine and Carbonyl group.
6.1 NEED FOR THE STUDY:
The literature survey reveals that sitagliptin is a novel drug for diabetes type 2 and only few methods has been reported for estimation ofSitagliptin phosphate in pharmaceutical formulation by Spectrophotometric and RP-HPLC. It reduces glycemia and glycogen level and is a potent drug for type 2 diabetes. Sensitive liquid chromatography tandem mass spectrometry method for the quantification of sitagliptin, a DPP-4 inhibitor, in human plasma using liquid-liquid extraction is been performed and stability indicating methods are also need to be developed for its invitro determination in gastric and intestinal fluids. Analysis of Sitagliptin in human urine using turbulent flow online extraction and tandem mass spectroscopy is also done. Molecular modeling and X-ray crystallography is been applied to design novel, potent sitagliptin. In the view of the need in the industry for routine analysis ofSitagliptin, attempts are being made to develop simple and accurate instrumental methods for quantitative estimation of sitagliptin. Thus there is need for the development of new, simple, sensitive and accurate analytical methods for the quantitative estimation of sitagliptin as an active pharmaceutical ingredient in bulk and pharmaceutical formulation .
6.2 REVIEW OF LITERATURE:

David Q et al.[1] have reported characterization of two cyclic metabolites of Sitagliptin.

  • Zeng Wei et al.[2] have reported Sitagliptin in human urine and hemodialysate using turbulent flow online extraction and tandem mass spectroscopy.
  • PK Sahoo et al.[3] have reported Simultaneous estimation of metformin hydrochloride and pioglitazone hydrochloride by HPTLC method from combined tablet dosage form.
  • Kyoung Soo Lim et al.[4] have reported Pharmacokinetics, pharmacodynamics, and tolerability of the dipeptidyl peptidase IV inhibitor LC15-0444 in healthy Korean men: A dose—block-randomized, double-blind, placebo-controlled, ascending single-dose, phase I study.
  • Ramakrishna Nirogi et al.[5] have reported Sensitive liquid chromatography tandem mass spectrometry method for the quantification of sitagliptin, a DPP-4 inhibitor, in human plasma using liquid-liquid extraction.
  • Tesfaye Biftu et al.[6] have reported Rational design of a novel, potent, and orally bioavailable cyclohexylamine DPP-4 inhibitor by application of molecular modeling and X-ray crystallography of Sitagliptin.

7 / 6.3 OBJECTIVES OF THE STUDY:
In the proposed work attempts shall be made.
  • To establish sensitive and accurate method/s for the quantitative estimation of DPP-4 inhibitor sitagliptin in bulk and dosage form.
  • To validate the newly developed methods in accordance with the analytical parameters mentioned in the USP/FDA guidelines.
  • To apply the newly developed, validated analytical methods for quantitative estimation of DPP-4 inhibitor sitagliptin in bulk and pharmaceutical dosage forms.
7.1 MATERIALS AND METHODS:
MATERIALS:
  • All the chemical and reagents for the development of new analytical method to estimate Sitagliptin phosphate will be procured from standard companies.
  • The method will be standardized on double beam UV-Visible spectrophotometer and or HPLC instruments using calibrated volumetric glass apparatus.
  • The pure sample of Sitagliptin for the research work will be procured from MERCK AND CO Pharmaceutical Industry, MUMBAI.
METHODS:
New method will be planed depending up on the chemical nature
(functional group.) of Sitagliptin.
For RP-HPLC various combinations and different ratio of solvent will be tried for selection of mobile phase for obtaining good resolution. By using retention time and chromatogram of Sitagliptin, a typical analytical parameters such as linearity, range, precision, specificity, accuracy, ruggedness and robustness used in assay validation will be determined.
7.2 SOURCE OF DATA:
  1. IISC library, Bangalore.
  2. SACCollege of pharmacy library, BG Nagara.
  3. RGUHS library, Bangalore.
  4. E-library, SACCollege of pharmacy.

8 / 7.3 METHOD OF COLLECTION OF DATA:
DATA COLLECTED FROM
JOURNALS:
  1. Journal of Pharmaceutical Research.
  2. India Journal of Pharmaceutical Sciences.
  3. Indian Journal of Pharmaceutical Education and Research.
  4. Journal of Chromatography.
  5. Asian Journal of Chemistry.
  6. Journal of Pharmaceutical and Biomedical Analysis.
RELATED LINKS:






7.4 DOES THE STUDY REQUIRE ANY INVESTIGATION OR INTERVENTIONS TO BE CONDUCTED ON PATIENTS OR OTHER HUMANS OR ANIMALS?
NOT APPLICABLLE.
7.5 HAS ETHICAL CLEARENCE BEEN OBTAINED FROM YOUR INSTITUTION IN CASE OF 7.4?
NOT APPLICABLE
REFERANCES:
1. David Q. Liu, Byron H. Arison, Ralph A. Stearns, Dooseop Kim, and Stella H. Vincent.Characterization of Two Cyclic Metabolites of Sitagliptin.
Drug metabolism and disposition35, 521, (2007)
2. Zeng. Wei, Musson. Donald G, Fisher. Alison L, Chen. Li, Schwartz. Michael S, Woolf. Eric J and Wang Amy Qiu. Determination of sitagliptin in human urine and hemodialysate using turbulent flow online extraction and tandem mass spectrometry. Journal of pharmaceutical and biomedical analysis46(3), 534, (2008).
3.PK Sahoo, R Sharma and SC Chaturvedi.
Simultaneous estimation of metformin hydrochloride and pioglitazone hydrochloride by RPHPLC method from combined tablet dosage form.
Indian journal of pharmaceutical science70(3), 383, (2008).
4. Kyoung Soo Lim , Jung-Ryul Kim , Yun-Jung Choi, Kwang-Hee Shin,Kyu-Pyo Kim , Jang-Hee Hong , Joo-Youn Cho, Hyun-Suk Shin, Kyung-Sang Yu, Sang-Goo Shin, O Hwan Kwon , Dal-Mi Hwang, Jeong-Ae Kim and In-Jin Jang.
Pharmacokinetics, pharmacodynamics, and tolerability of the dipeptidyl peptidase IV inhibitor LC15-0444 in healthy Korean men: A dose—block-randomized, double-blind, placebo-controlled, ascending single-dose, phase I study.
Clinical therapeutics30(10), 1817, (2008).
5. Ramakrishna Nirogi, Vishwottam Kandikere, Koteshwara Mudigonda, Prashanth Komarneni, Raghupathi Aleti and Rajeshkumar Boggavarapu.
Sensitive liquid chromatography tandem mass spectrometry method for the quantification of sitagliptin, a DPP-4 inhibitor, in human plasma using liquid-liquid extraction. Biomedical chromatography22(10), 1817, (2008).
6. Tesfaye Biftu, Giovanna Scapin, Suresh Singh, Dennis Feng, Joe W. Becker, George Eiermann, Huaibing He, Kathy Lyons, Sangita Patel, Aleksandr Petrov, Ranabir Sinha-Roy, Bei Zhang, Joseph Wu, Xiaoping Zhang, George A. Doss, Nancy A. Thornberry and Ann E. Weber.
Rational design of a novel, potent, and orally bioavailable cyclohexylamine DPP-4 inhibitor by application of molecular modeling and X-ray crystallography of Sitagliptin.
Bioorganic and medicinal chemistry letters17(12), 3384,(2007).
7. Mi Ae Jun, Woul Seong Park, Seung Kyu Kang, Ki Young Kim, Kwang Rok Kim, Sang Dal Rhee, Myung Ae Bae, Nam Sook Kang, Sang-Kwon Sohn, Sung Gyu Kim, Jie Oh Lee, Duck Hyung Lee, Hyae Gyeong Cheon, Sung Soo Kim and Jin Hee Ahn.
Synthesis and biological evaluation of pyrazoline analogus with b amino acyl group synthesis as dipeptidyl peptidase 4 inhibitors.
EuropeanJournal of Medicinal Chemistry43(9), 1889, (2008).
8.Maria G. Beconi, James R. Reed, Yohannes Teffera, Yuan-Qing Xia, Christopher J. Kochansky, David Q. Liu, Shiyao Xu, Charles S. Elmore, Suzanne Ciccotto, Donald F. Hora, Ralph A. Stearns and Stella H. Vincent
Disposition of the Dipeptidyl Peptidase 4 Inhibitor Sitagliptin in Rats and Dogs.
Drug Metabolism and Disposition Fast Forward1(1), 525, (2007).
9. Raz I, Hanefeld M, Xu L, et al. Efficacy and safety of the dipeptidyl peptidase–4 inhibitor sitagliptin as monotherapy in patients with type 2 diabetes mellitus. Diabetologia.49,2564, (2006).
10.Bo Ahren, Mona Landin-Olsson, Per-Anders Jansson, Maria Svensson, David Holmes and Anja Schweizer
Inhibition of Dipeptidyl Peptidase-4 Reduces Glycemia, Sustains Insulin Levels, and Reduces Glucagon Levels in Type 2 Diabetes.
The Journal of Clinical Endocrinology & Metabolism89(5), 2078, (2004).
11. Gary a Herman, Arthur Bergman, Bingming Yi and mark Kipness.
Tolerability and pharmacokinetics of metformin and the DPP 4 inhibitor Sitagliptin when co administered in patients with type 2 diabetes.
Current medical research and opinion22(10), 1939, (2006).
9 / Signature of the candidate / PATIL .S. L
10 / Remarks of the guide / The proposed research work is original and designed on rational basis. It would be a good contribution.
11 / 11.1 Name and Designation of Guide
Guide ship reference No. Of RGUHS
11.2 Signature / Dr.B.RAMESH
M.Pharm Ph.D
Professor
Department of Pharmaceutical chemistry
S.A.C. college of pharmacy
B.G.Nagara 571448
Karnataka
11.3 Co-Guide
11.4 Signature / NOT APPLICABLE
11.5 Head of the Department
11.6 Signature / Dr. B.RAMESH
M.Pharm. Ph.D.
Professor & Head,
Department of Pharmaceutical chemistry
S.A.C.College of Pharmacy,
B.G.nagara. 571448
Karnataka
12 / 12.1Remarks of the Principal
12.2 Signature