Targeting the p53/MDM2 protein-protein interaction

Goffin, Sarah Anne (2016) Targeting the p53/MDM2 protein-protein interaction. Doctoral thesis, University of East Anglia.

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Abstract

The p53/MDM2 protein-protein interaction is the most widely characterised proteinprotein interaction to date. As of 2014, there are over 20 compounds that have been shown to the p53-MDM2 protein-protein interaction, however many compounds have not progressed into clinical trials due to their high hydrophobicity.
Herein we describe the synthesis, molecular modelling, physical characterisation and biological testing of novel inhibitors of the p53/MDM2 protein-protein interaction based on the natural product chlorofusin.
The first focus is a combinatorial library generated in the Searcey laboratory of known p53/MDM2 protein-protein interaction inhibitors with the desire to generate novel analogues and study their interactions with the protein through NMR spectroscopy and molecular modelling. These compounds were tested by in a fluorescence polarisation assay and also in cell lines overexpressing MDM2 as well as p53-null cells as a comparator. This generated two novel compounds shown to have activity selectively for the p53/MDM2 protein-protein interaction.
The second chapter focuses on simplified substitutions of the azaphilone (the chromophore portion of chlorofusin, a natural product inhibitor of the p53-MDM2 proteinprotein interaction): initially with simple fused bicyclic carboxylic acids and later using click chemistry substitutions. Interestingly, in vitro studies showed that the click
analogues retained activity or activity improved when the peptide portion was removed and hence further studies of the click amino acid analogues were generated. This library generated one analogue that was active in vitro as well as selectively in MDM2-overexpressing cell lines.
The third chapter focusses on the azaphilone chromophore present in the natural product chlorofusin. The Sonogashira precursor used to generate azaphilone analogues was synthesised using a methodology adopted by Porco et al and subsequent analogues were generated using a novel double-Sonogashira approach followed by functionalisation published by Boger et al. Once the azaphilone was synthesised, metholodogies were trialled in order to condense the azaphilone with the chlorofusin peptide in order to create analogues containing both the peptide and small molecule portions of chlorofusin.
In addition, molecular modelling was attempted to generate novel binding analogues.

Item Type: Thesis (Doctoral)
Faculty \ School: Faculty of Science > School of Pharmacy
Depositing User: Brian Watkins
Date Deposited: 10 Mar 2016 14:27
Last Modified: 10 Mar 2016 14:27
URI: https://ueaeprints.uea.ac.uk/id/eprint/57422
DOI:

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