Structure- and Reactivity-Based Development of Covalent Inhibitors of the Activating and Gatekeeper Mutant Forms of the Epidermal Growth Factor Receptor (EGFR)

Ward, Richard A.; Anderton, Mark; Ashton, Susan; Bethel, Paul A.; Box, Matthew; Butterworth, Sam; Colclough, Nicola; Chorley, Christopher G.; Chuaqui, Claudio; Cross, Darren A.E.; Dakin, Les A.; Debreczeni, J.; Eberlein, Cath; Finlay, M. Raymond V.; Hill, George B.; Grist, Matthew; Klinowska, Teresa; Lane, Clare; Martin, Scott; Orme, Jonathon P.; Smith, Peter D.; Wang, Fengjiang; Waring, Michael J.

doi:10.1021/jm400822z

Cited by 217 publications

(234 citation statements)

References 44 publications

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“…reducing K inact ) whilst retaining overall inhibitory activity. 23 Substitution of the maleimide double bond offered the possibility of modulating the electrophilicity of the Michael acceptor warhead, once additional non-covalent interactions had been identified.…”

Section: Irreversible Binders At Hras:sos Site Cmentioning

confidence: 99%

Small Molecule Binding Sites on the Ras:SOS Complex Can Be Exploited for Inhibition of Ras Activation

et al. 2015

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Constitutively active mutant KRas displays a reduced rate of GTP hydrolysis via both intrinsic and GTPaseactivating protein-catalysed mechanisms, resulting in the perpetual activation of Ras pathways. We describe a fragment screening campaign using X-ray crystallography that led to the discovery of three fragment binding sites on the Ras:SOS complex. The identification of tool compounds binding at each of these sites allowed exploration of two new approaches to Ras pathway inhibition by stabilising or covalently modifying the Ras:SOS complex to prevent the reloading of Ras with GTP. Initially, we identified ligands that bound reversibly to the Ras:SOS complex in two distinct sites, but these compounds were not sufficiently potent inhibitors to validate our stabilisation hypothesis. We conclude by demonstrating that covalent modification of Cys118 on Ras leads to a novel mechanism of inhibition of the SOS-mediated interaction between Ras and Raf, and is effective at inhibiting the exchange of labelled GDP in both mutant (G12C and G12V) and wild type Ras.

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Section: Irreversible Binders At Hras:sos Site Cmentioning

confidence: 99%

Small Molecule Binding Sites on the Ras:SOS Complex Can Be Exploited for Inhibition of Ras Activation

et al. 2015

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“…Recently, this interaction was shown to have a dramatic effect on inhibitory activity and mutant-selectivity in case of various EGFR inhibitors. 39,40 Synthesis of a Focused Small Molecule Library.…”

Section: Rational Design Of Egfr Inhibitors Based On Screening Hit Anmentioning

confidence: 99%

Targeting Drug Resistance in EGFR with Covalent Inhibitors: A Structure-Based Design Approach

et al. 2015

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Receptor tyrosine kinases represent one of the prime targets in cancer therapy, as the dysregulation of these elementary transducers of extracellular signals, like the epidermal growth factor receptor (EGFR), contributes to the onset of cancer, such as non-small cell lung cancer (NSCLC). Strong efforts were directed to the development of irreversible inhibitors and led to compound CO-1686, which takes advantage of increased residence time at EGFR by alkylating Cys797 and thereby preventing toxic effects. Here, we present a structure-based approach, rationalized by subsequent computational analysis of conformational ligand ensembles in solution, to design novel and irreversible EGFR inhibitors based on a screening hit that was identified in a phenotype screen of 80 NSCLC cell lines against approximately 1500 compounds. Using protein X-ray crystallography, we deciphered the binding mode in engineered cSrc (T338M/S345C), a validated model system for EGFR-T790M, which constituted the basis for further rational design approaches. Chemical synthesis led to further compound collections that revealed increased biochemical potency and, in part, selectivity toward mutated (L858R and L858R/T790M) vs nonmutated EGFR. Further cell-based and kinetic studies were performed to substantiate our initial findings. Utilizing proteolytic digestion and nano-LC-MS/MS analysis, we confirmed the alkylation of Cys797.

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“…Recently, an irreversible inhibitor, WZ4002, was described that showed mutant-selective activity in preclinical models but did not progress to human clinical trials (22). Other irreversible and reversible EGFR mutant-selective inhibitors have recently been described (23)(24)(25). Through structure-based design we identified 4 irreversible inhibitors that are EGFR mutant-selective.…”

Section: Introductionmentioning

confidence: 99%

In Vitro and In Vivo Characterization of Irreversible Mutant-Selective EGFR Inhibitors That Are Wild-Type Sparing

Sjin

Lee

Walter

et al. 2014

Molecular Cancer Therapeutics

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Patients with non-small cell lung carcinoma (NSCLC) with activating mutations in epidermal growth factor receptor (EGFR) initially respond well to the EGFR inhibitors erlotinib and gefitinib. However, all patients relapse because of the emergence of drug-resistant mutations, with T790M mutations accounting for approximately 60% of all resistance. Second-generation irreversible EGFR inhibitors are effective against T790M mutations in vitro, but retain affinity for wild-type EGFR (EGFR WT

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Structure- and Reactivity-Based Development of Covalent Inhibitors of the Activating and Gatekeeper Mutant Forms of the Epidermal Growth Factor Receptor (EGFR)

Cited by 217 publications

References 44 publications

Small Molecule Binding Sites on the Ras:SOS Complex Can Be Exploited for Inhibition of Ras Activation

Small Molecule Binding Sites on the Ras:SOS Complex Can Be Exploited for Inhibition of Ras Activation

Targeting Drug Resistance in EGFR with Covalent Inhibitors: A Structure-Based Design Approach

In Vitro and In Vivo Characterization of Irreversible Mutant-Selective EGFR Inhibitors That Are Wild-Type Sparing

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