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dc.contributor.advisorHoelder S
dc.contributor.authorDarlow, W
dc.contributor.editorHoelder, S
dc.date.accessioned2022-12-14T14:09:01Z
dc.date.available2022-12-14T14:09:01Z
dc.date.issued2022-12-12
dc.identifier.citation2022en_US
dc.identifier.urihttps://repository.icr.ac.uk/handle/internal/5608
dc.description.abstractChemically induced degradation has emerged as a valuable tool in chemical biology and medicinal chemistry for both potential therapies, and to investigate protein function. Proteolysis targeting chimeras (PROTACs) are small molecules that can hijack E3 ligases, the protein complexes that catalyse ubiquitination, to direct proteasomal degradation of selected targets. Simultaneous binding of both the E3 ligase recruiting moiety and warhead to their respective targets induces ternary (E3-PROTAC-target) complex formation, allowing the E3 ligase to perform ubiquitination on non-native substrates. Subsequent polyubiquitination of the target protein signals for proteasome dependant degradation. Despite significant strides in PROTAC development, including multiple early phase clinical trials, currently only proteins with known binders can be targeted for degradation. We hypothesise that to improve the target landscape of PROTACs and increase the degradable proteome, we could implement a proteomics-based screen of promiscuous kinase focussed PROTACs. An array of small molecular weight kinase binding PROTACs were synthesised and triaged through proteomics analysis, identifying selective NEK9 and Aurora kinase A (AURKA) hit degraders. Subsequent validation confirmed degradation of NEK9, however, through a neddylation and CRBN independent mechanism. Warhead alterations were explored to determine what component of the degrader conferred the unexpected degradation mechanism. A selective AURKA PROTAC was found to be active at low nM concentrations, affording robust reductions in protein levels over a 2-72 hour treatment. Warhead substitutions to previously published AURKA inhibitors in addition to truncation of the warhead afforded reduced degradation potency. Through the use of small molecular weight warheads, we highlight the ternary complex driven degradation or AURKA, and demonstrate that some PROTACs may not require high affinity warheads. Our approach has the potential to afford degraders of underexplored proteins and provide valuable chemical tools to elucidate protein function.
dc.language.isoengen_US
dc.publisherInstitute of Cancer Research (University Of London)en_US
dc.rights.urihttps://www.rioxx.net/licenses/all-rights-reserveden_US
dc.titleDiscovery of NEK9 and Aurora A PROTACs using a proteomics-based screening approachen_US
dc.typeThesis or Dissertation
dcterms.accessRightsPublic
dc.date.updated2022-12-14T14:06:15Z
rioxxterms.versionAOen_US
rioxxterms.licenseref.urihttps://www.rioxx.net/licenses/all-rights-reserveden_US
rioxxterms.licenseref.startdate2022-12-12
rioxxterms.typeThesisen_US
pubs.organisational-group/ICR
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Cancer Therapeutics
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Cancer Therapeutics/Medicinal Chemistry 4 (including Analytical Chemistry)
pubs.organisational-group/ICR/Students
pubs.organisational-group/ICR/Students/PhD and MPhil
pubs.organisational-group/ICR/Students/PhD and MPhil/18/19 Starting Cohort
icr.researchteamMedicinal Chemistry 4en_US
dc.contributor.icrauthorDarlow, William
uketdterms.institutionInstitute of Cancer Research
uketdterms.qualificationlevelDoctoral
uketdterms.qualificationnamePh.D
icr.provenanceDeposited by Mr Barry Jenkins (impersonating Mr Will Darlow) on 2022-12-14. Deposit type is initial. No. of files: 1. Files: Thesis With Corrections Will Darlow.pdf
dc.type.qualificationlevelDoctoral
dc.type.qualificationnamePh.D


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