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Synthesis and Evaluation of Novel 20-HETE formation inhibitors

Deshpande, Ameya (2018) Synthesis and Evaluation of Novel 20-HETE formation inhibitors. Master's Thesis, University of Pittsburgh. (Unpublished)

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Abstract

20-hydroxyecosatetraenoic acid (20-HETE) is a metabolite of arachidonic acid (AA) formed via CYP4 enzymes and has been shown to have strong vasoconstriction activity in the microvasculature. Epoxyecosatetraenoic acids (EETs) products of the epoxygenation pathway and possess vasodilatory function. Both of these fatty acid metabolites have demonstrated opposite effects in the microvasculature. 20-HETE has been studied in-vitro and in-vivo with results suggesting a role in secondary brain injury after subarachnoid hemorrhage (SAH) or cardiac arrest (CA). Several molecules, which target the formation of 20-HETE, have been studied and reported in literature such as 1-ABT, 17-ODYA and others. There has been no clinical translation for any of them due to their physicochemical properties, potency and selectivity. Over 15 years ago Taisho disclosed two molecules, both showing excellent potency and selectivity towards 20-HETE-formation inhibition over EET formation. However, these compounds were marred by chemical instability at low pH, poor solubility and a shockingly low T1/2. Currently, there are no CYP4 inhibitors in the clinic for the purpose of neuroprotection. Through scaffold hopping of compound 8, a 20-HETE formation inhibitor with poor metabolic stability, novel leads UPMP00010/19 were designed in McDermott lab. These leads have improved stability when compared to compound 8. Our aim is to further optimize these leads by the synthesis of derivatives with physicochemical properties appropriate for compounds that must act in the brain. As a part of greater optimization effort, we synthesized 24 UPMP00010/19 derivatives and these compounds are the focus of this work. Compounds generated were tested against a panel of enzymes, which include human liver microsomes (HLM), recombinant CYP4F2 (rCYP4F2), rat liver microsomes (RLM) and rat kidney microsomes (RKM) in the lab of Dr. Samuel Poloyac at the University of Pittsburgh. Compounds with promising potency were taken for further evaluation, which included determination of intrinsic clearance, kinetic solubility and CNS permeability potential. Compounds 21a showed promising potency (IC50=73nM), good HLM stability (92% parent molecule remaining at 30 mins) and high CNS permeability potential (efflux ratio=0.803). Compound 21c is equipotent (IC50=79nM) to 21a and has good HLM stability (92% parent molecule remaining at 30 mins). 21a showed very little inhibition of EET formation at the highest concentration tested (3.1% @ 50,000nM). Compounds 21f and 21h are particularly attractive molecules as these molecules have excellent results in single point titrations in both human and rat enzyme systems.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Deshpande, Ameyaaud10@pitt.eduaud10
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairMcDermott, Leelam179@pitt.edulam179
Committee MemberIyer, Premapci2@pitt.edupci2
Committee MemberFolan, Maggiefolanm@pitt.edufolanm
Date: 31 July 2018
Date Type: Publication
Defense Date: 23 July 2018
Approval Date: 31 July 2018
Submission Date: 26 July 2018
Access Restriction: 5 year -- Restrict access to University of Pittsburgh for a period of 5 years.
Number of Pages: 135
Institution: University of Pittsburgh
Schools and Programs: School of Pharmacy > Pharmaceutical Sciences
Degree: MS - Master of Science
Thesis Type: Master's Thesis
Refereed: Yes
Uncontrolled Keywords: Keywords: 20-HETE, Cardiac arrest, subarachnoid hemorrhage, Vasoconstriction, Scaffold-hopping, human liver microsomes, rat liver microsomes, rat kidney microsomes CNS permeability, Kinetic solubility, HLM stability.
Date Deposited: 31 Jul 2018 12:18
Last Modified: 31 Jul 2018 12:18
URI: http://d-scholarship.pitt.edu/id/eprint/35025

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