Understanding Selectivity In Irreversible FGFR4 Inhibition

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dc.contributor.advisor Squire, C en
dc.contributor.advisor Smaill, J en
dc.contributor.author Lin, Xiaojing en
dc.date.accessioned 2018-06-07T04:21:10Z en
dc.date.issued 2018 en
dc.identifier.uri http://hdl.handle.net/2292/37227 en
dc.description Full text is available to authenticated members of The University of Auckland only. en
dc.description.abstract The fibroblast growth factor receptor family (FGFR1-4) regulates cellular development and metabolism. Aberrant FGFR signalling is a major driving force in various malignancies, and therefore, represents a promising therapeutic target. FGFR4 aberrations are implicated in driving hepatocellular carcinoma (HCC) and much research effort has been invested into developing FGFR4 inhibitors. However, poor target selectivity remains a great hurdle in kinase inhibitor development owing to the highly conserved ATP-binding site present across the kinome. Selectively targeting FGFR4 while sparing the closely related FGFR1-3 is particularly difficult. Another limiting factor for the clinical success of kinase inhibitors is the development of gatekeeper-mutation-associated drug resistance. BLU9931 was the first reported irreversible FGFR4-selective inhibitor, its successor, BLU554, is currently under Phase I clinical trial in HCC patients. Based on BLU9931, our collaborators developed a series of inhibitors with several modifications including increased flexibility. This project aimed to investigate and compare our inhibitor series with BLU9931 and BLU554 at the molecular level, to expand our understanding of selective FGFR4-inhibition, and to provide the data to help overcome potential gatekeeper-associated drug resistance. To probe the selectivity profiles of the inhibitor series, differential scanning fluorimetry (DSF) and liquid chromatography-mass spectrometry (LC-MS) were employed. DSF results were confounded by the covalent reactions between FGFR4 and the inhibitors. However, DSF represents a high-throughput strategy to detect off-target binding. LC-MS experiments, more than 160 in number, not only confirmed the covalent binding and stoichiometry of the irreversible inhibitors binding to FGFR4 but provided information on their relative reactivity towards the FGFR family and FGFR4 gatekeeper variants. Six crystal structures were determined and revealed detailed binding modes of the inhibitors and a novel mechanism of off-target activity. This project reports that the modifications within our inhibitor series provide advantages over BLU9931 in both FGFR4-selectivity and in accommodating gatekeeper mutations. However, BLU554 appears superior in both regards. In addition, a set of tools and protocols are now in place in our laboratories such that newly developed irreversible kinase inhibitors, either our own or developed by our collaborators, can be rapidly characterised in vitro helping to progress these molecules towards human clinical trial. en
dc.publisher ResearchSpace@Auckland en
dc.relation.ispartof Masters Thesis - University of Auckland en
dc.relation.isreferencedby UoA99265076513302091 en
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. en
dc.rights Restricted Item. Available to authenticated members of The University of Auckland. en
dc.rights.uri https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm en
dc.rights.uri http://creativecommons.org/licenses/by-nc-sa/3.0/nz/ en
dc.title Understanding Selectivity In Irreversible FGFR4 Inhibition en
dc.type Thesis en
thesis.degree.discipline Biological Science en
thesis.degree.grantor The University of Auckland en
thesis.degree.level Masters en
dc.rights.holder Copyright: The author en
pubs.elements-id 744128 en
pubs.org-id Medical and Health Sciences en
pubs.org-id Medical Sciences en
pubs.org-id Auckland Cancer Research en
pubs.record-created-at-source-date 2018-06-07 en


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