“QUANTITATIVE ESTIMATION AND VALIDATION OFPRASUGREL HYDROCHLORIDE IN PHARMACEUTICAL FORMULATIONS”

DISSERTATION PROTOCOL

SUBMITTED TO

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

BANGALORE, KARNATAKA.

BY

RAVIKIRAN SHANKAR MARKAD

M.PHARM, PART-I

DEPARTMENT OF QUALITY ASSURANCE

NARGUND COLLEGE OF PHARMACY

BANGALORE-8

(2010-2012)

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) / RAVIKIRAN SHANKAR MARKAD
NARGUND COLLEGE OF PHARMACY,
DATTATREYANAGAR, IInd MAIN,
100 FEET RING ROAD,
BSK IIIrd STAGE,
BANGALORE-85,
KARNATAKA.
2. /

NAME OF THE INSTITUTION

/ NARGUND COLLEGE OF PHARMACY,
DATTATREYANAGAR, IInd MAIN,
100 FEET RING ROAD,
BSK IIIrd STAGE,
BANGLORE-85,
KARNATAKA.
3. /

COURSE OF STUDY AND SUBJECT

/ MASTER OF PHARMACY IN
QUALITY ASSURANCE
4. / DATE OF ADMISSION OF COURSE / 1ST JULY 2010.
5. /

TITLE OF TOPIC

/ “QUANTITATIVE ESTIMATION AND VALIDATION OFPRASUGREL HYDROCHLORIDE IN PHARMACEUTICAL FORMULATIONS”
6.
6.1
6.2 / Brief resume of the intended work:
Need for the study:
Prasugrel is an oral anti-platelet medicine specifically for patients who have undergone an angioplasty procedure to open up a blocked heart artery after experiencing a heart attack or heart-related chest pain at rest (unstable angina) – both these medical conditions are known as acute coronary syndrome. Prasugrel is a member of the thienopyridine class of ADP receptor inhibitor ofplateletactivation and aggregation mediated by the P2Y12 ADP receptor [1].These agents reduce the aggregation ("clumping") of platelets by irreversibly binding to P2Y12 receptors [2].
PrasugrelHCl is available in the form of tablet in different doses 5mg and 10mg.There is no analytical method reported for quantitative estimation ofPrasugrel HCL in pharmaceutical dosage form. Hence, there is a need to develop simple, accurate, precise, sensitive analytical method for Prasugrel HCL in tablet dosage form.
Review of literature:
Chemically Prasugrel HCL designated as 5-[(1RS)-2-cyclopropyl-1-(2-fluorophenyl)-2-oxoethyl]-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl acetate hydrochloride.
Molecular formula- C20H20FNO3S.HCl.
Molecular weight - 409.90 g/mol.
Characteristic - Prasugrel hydrochloride is a white to practically white solid.
Chemical structure of Prasugrel hydrochloride.

Solubility - It dissolves freely in methanol and is slightly soluble in 1- and 2-propanol and acetone. It is practically insoluble in diethyl ether and ethyl acetate [1].
Wickremsinhe ER, Tian Y, Ruterbories KJ, Verburg EM, Weerakkody GJ, Kurihara A et al studied stereoselective metabolism of Prasugrel in humans using a novel chiral liquid chromatography-tandem mass spectrometer after derivatization with bromomethoxyacetophenone to stabilize the molecule [3].
Wong PC, Crain EJ, Watson CA, Hua J, Schumacher WA, Rehfuss R studied effects of thrombosis, haemostasis, platelet function and response variability of clopidogrel and Prasugrel in rabbits. The goal of study was whether induced platelet aggregation (IPA) level or P2Y(12) receptor occupancy (RO) could be optimized to better balance the efficacy and bleeding effects of these thienopyridines and reduce the response variability in rabbits [4].
Farid NA, McIntosh M, Garofolo F, Wong E, Shwajch A, Kennedy Met al developed fast and sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS)-based bioanalytical assays for determination of the active and inactive metabolites of Prasugrel in human plasma. After extraction and separation, the analytes were detected and quantified using a triple quadrupole mass spectrometer using positive electrospray ionization [5].
Mousa SA, Jeske WP, Fareed J studied novel adenosine diphosphate (ADP) P2Y12 antagonists, including Prasugrel, ticagrelor, cangrelor and elinogrel. The ADP P2Y12 antagonists have advantages over clopidogrel ranging from faster onset to greater and less variable inhibition of platelet function. Novel ADP P2Y12 antagonists are under investigation to determine whether their use can result in improved antiplatelet activity, faster onset of action, and/or greater antithrombotic effects than clopidogrel, without an unacceptable increase in hemorrhagic or other side effects [6].
K. Hagihara, M. Kazui, H. Ikenaga, T. Nanba, K. Fusegawa, M. Takahashi et al comparatively studied the efficiency of the formation of Prasugrel and clopidogrelthiolactones and their active metabolitesin rats and dogs.The areas under the plasma concentration versus time curve (AUC) of the thiolactone intermediates in the portal vein plasma after an oral dose of Prasugrel (1 mg kg−1) and clopidogrel (0.77 mg kg−1) respectively, in rats, respectively, in dogs, indicating efficient hydrolysis of Prasugrel and little metabolism of clopidogrel to their thiolactones in the intestine [7].
Smith RL, Gillespie TA, Rash TJ, Kurihara A, Faridto NA estimateddisposition and metabolic fate of Prasugrel in mice, rats, and dogs of Prasugrel.Prasugrel was rapidly absorbed and extensively metabolized. In the mouse and dog, maximum plasma concentration of radioactivity was observed in less than 1h after an oral Prasugrel dose. Most of the administered Prasugrel dose was recovered in the faeces of rats and dogs (72% and 52–73%, respectively), and in mice urine (54%) [8].
Testa L, Bhindi R, Van Gaal WJ, Latini RA, PizzocriS, Lanotte S, BiondiZoccai GL, Valgimigli M, Laudisa ML, Brambilla N et al
worked on the risk of intensifying platelet inhibition beyond clopidogrel? systematic review and a critical appraisal of the role of Prasugrel.
has been published recently, Prasugrel has been developed and tested in several ex vivo studies and clinical trials showing able to provide a more powerful antiplatelet effect at the expense of a higher risk of bleeding complications [9].
Lazar LD, Lincoff AM claim that Prasugrel is more potent, faster in onset, and more consistent in inhibiting platelets.Prasugrel (Effient) has been approved for reducing the risk of thrombotic complications in patients with acute coronary syndromes who are to undergo percutaneous coronary intervention [10].
Williams ET, Jones KO, Ponsler GD, Lowery SM, Perkins EJ, Wrighton SA et al studied biotransformation of Prasugrel, by the Human Carboxylesterases 1 and 2.The biotransformation of Prasugrel to its active metabolite, requires ester bond hydrolysis, forming the thiolactone, followed by cytochrome P450–mediated metabolism to the active metabolite. The presumed role of the human liver- and intestinal-dominant carboxylesterases, hCE1 and hCE2, respectively, in the conversion of Prasugrel to R-95913 was determined using expressed and purified enzymes[11].
Borole TC, Mehendre R, Damle MC, Bothara KG development and validation of stability indicating HPTLC method for determination of Prasugrel.The method employed TLCaluminium plates precoated with silica gel 60 F254 as the stationary phase. The solvent systemconsisted of Dichloromethane: Methanol (9.9:0.1v/v). This system was found to give compactspot for Prasugrel (Rf value 0.58±0.03). Prasugrel was subjected to stress test conditions likeacid, alkali, neutral hydrolysis, oxidation, dry heat and photo degradation. The spot for productof degradation was well resolved from the drug. Densitometric analysis of drug was carried outin the absorbance mode at 254 nm [12].
6.3
7.
7.1 / Objectives of the study:
Literature survey reveals that, there is no spectrophotometric method available for the estimation of PrasugrelHCl in bulk and various pharmaceutical dosage forms.
The objectives of the present work are,
  1. To develop analytical method for is PrasugrelHCl in bulk and various pharmaceutical dosage forms by UV-Visible spectrophotometer.
  2. To developandoptimizeHPLC method to estimate PrasugrelHClin pharmaceutical formulations.
  3. To validate all developed analytical methods as per ICH guidelines.
Materials and Methods
Materials:
PrasugrelHCl, PrasugrelHCl tablet, methanol and ethanol,distilled water, hexane, ethylacetate, dichloromethane,acetonitrile, glacial acetic acid,acetone,propanol,hydrochloric acid, sodium hydroxide, potassium dihydrogen phosphate, derivatization agents etc.
Source of data:
Data will be obtained from Google, Pubmed, Science Direct, Medline, and other internet facilities, literature search and related articles from library of NargundCollege of Pharmacy, Digital Library of RGUHS, Bangalore, etc.
Journals
Drug Metabolism and Disposition
Rapid Communication in Mass Spectroscopy Research Article
Clinical and Applied Thrombosis/Haemostats
Journal of Pharmacy Practice
QJM (Oxford Journal)
Cleveland Clinic Journal of Medicine
Internet Browsing
  • india.com
Text Books And official pharmacopoeia
  • Beckett AH, Stenlake JB. Practical pharmaceutical chemistry. 4th ed. Part II, Delhi: CBS publishers and distributors; 1997.
  • Sweetman SC. Martindale the complete drug reference. Martindale 34th ed. London,Chicago: Pharmaceutical Press;2005.
  • Sethi PD, Quantitative analysis of drugs in pharmaceutical formulations, 3rd ed. Delhi:CBS publishers and distributors.
  • Lawrence A. Tressel, Stability-indicating HPLC methods for drug analysis. 3rd ed.Pharmaceutical Press London, UK.

7.2
7.3
7.4 / Method Of Collection Of Data (Including sampling procedure,if any)
  1. Procurement of the drug sample and chemicals required.
  2. Determination of the solubility of PrasugrelHCl in various solvents and buffers.
  3. Scanning the solution on UV- Visible spectrophotometer and selecting the solvents for various analytical studies.
  4. Preparation of standard calibration curve in suitable solvent.
  5. Develop analytical method for PrasugrelHCl in bulk and various pharmaceutical dosage forms by UV-Visible spectrophotometer using suitable zero/first/second order and AUC.
  6. Development and optimization of HPLC method to estimate PrasugrelHCl in pharmaceutical formulations.
  7. Validation of all developed analytical methods as per ICH guidelines.
Does the study require any investigation or intervention to be conducted on patients or otherhumans? If so please mention briefly.
-NOT APPLICABLE-
Has ethical clearance been obtained from your institution in case of 7.3?
-NOT APPLICABLE-
8. / LIST OF References:
  1. (asses date Oct 28, 2010).
  2. Wiviott SD, Braunwald E, McCabe CH, et al.(2007).“Prasugrel versus clopidogrel in patients with acute coronary syndromes”. N Engl J Med 357(20): 2001-15.
[Available from:
  1. Wickremsinhe ER, Tian Y, Ruterbories KJ, Verburg EM, Weerakkody GJ, Kurihara A et al.Stereoselective metabolism of Prasugrel in humans using a novel chiral liquid chromatography-tandem mass spectrometer method. The American Society for Pharmacology and Experimental Therapeutics. 2007; 35(6):917-21.
[Available from:
  1. Wong PC,Crain EJ, Watson CA, Hua J, Schumacher WA, Rehfuss R. Clopidogrel versus Prasugrel in rabbits effects on thrombosis, haemostasis, platelet function and response variability.ThrombHaemost. 2009 Jan; 101(1):108-15.
[Available from:
  1. Farid NA, McIntosh M, Garofolo F, Wong E, Shwajch A, Kennedy Zet al. Determination of the active and inactive metabolites of Prasugrel in human plasma by liquid chromatography/tandem mass spectrometry. Rapid Communications in Mass Spectrometry. 2007 Jan 30; 21(2):169-79.
[Available from:
  1. Mousa SA, Jeske WP, and Fareed J. antiplatelet therapy Prasugrel: A novel platelet ADP P2Y12 receptor antagonist. Clinical and applied thrombosis/hemostasis. 2010 Apr 1; 16(2):170-6.
[Available from:
  1. Hagihara K, Kazui M, Ikenaga H, Nanba T, Fusegawa K, Takahashi M et al. Comparison of formation of thiolactones and active metabolites of Prasugrelandclopidogrel in rats and dogs. Informa healthcare-Xenobiotica. 2009 Mar; 39(3):218-26.
[
  1. Smith RL, Gillespie TA, Rash TJ, Kurihara A, Faridto NA. Disposition and metabolic fate of Prasugrel in mice, rats, and dogs. Informa healthcare-Xenobiotica.2007; 37(8):884-901.
[Available from:
  1. Testa L, Bhindi R, Van Gaal WJ, Latini RA, Pizzocri S, LanotteS, BiondiZoccai GL, ValgimigliM, Laudisa ML, Brambilla N et al.
    What is the risk of intensifying platelet inhibition beyond clopidogrel? A systematic review and a critical appraisal of the role of Prasugrel.
    QJM.2010; 103(6):367-77.
[ Available from:
  1. Lazar LD, LincoffI AM. Prasugrel for acute coronary syndromes: Faster, more potent, but higher bleeding risk. CCJM.2009; 76(12):707-14.
[Available from:
  1. Williams ET, Jones KO, Ponsler GD, Lowery SM, Perkins EJ, Wrighton SA et al. The Biotransformation of Prasugrel, a New ThienopyridineProdrug, by the Human Carboxylesterases 1 and 2. DMD.2008; 36(7):1227-32.
[Available from:
  1. Borole TC, Mehendre R, Damle MC, Bothara KG. Development and validation of stability indicating HPTLC method for determination of Prasugrel.JCPR.2010;2(4):907-13.
[Available from:
9. / Signature of the candidate / (RAVIKIRAN SHANKAR MARKAD)
10. / Remarks of the Guide / RECOMMENDED FOR THE DISSERTATION WORK.
11. / Name & Designation of
(in block letters)
11.1Guide
11.2 Signature / DR. L.V. G. NARGUND PROFESSOR, HEAD OF THE DEPARTMENT, DEPARTMENT OF PHARMACEUTICAL CHEMISTRY NARGUND COLLEGE OF PHARMACY, BANGALORE
11.5 Head of the department
11.6 Signature / DR.J.N. NARENDRASHARATHCHANDRA HEAD OF THE DEPARTMENT, DEPARTMENT OF QUALITY ASSURANCENARGUND COLLEGE OF PHARMACY, BANGALORE
12. / 12.1 Remarks of Principal
12.2 Signature. / FORWARDED AND RECOMMENDED FOR FAVOURABLE CONSIDERATION.