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. 2019 Aug 2;10(9):1322-1327.
doi: 10.1021/acsmedchemlett.9b00276. eCollection 2019 Sep 12.

Structural and Molecular Insight into Resistance Mechanisms of First Generation cMET Inhibitors

Gavin W Collie  1 , Cheryl M Koh  2 , Daniel J O'Neill  1 , Christopher J Stubbs  1 , Puneet Khurana  1 , Alice Eddershaw  1 , Arjan Snijder  3 , Fredrik Mauritzson  3 , Louise Barlind  3 , Ian L Dale  1 , Joseph Shaw  1 , Christopher Phillips  1 , Edward J Hennessy  4 , Tony Cheung  4 , Ana J Narvaez  1
Affiliations

Structural and Molecular Insight into Resistance Mechanisms of First Generation cMET Inhibitors

Gavin W Collie et al. ACS Med Chem Lett. .

Abstract

Many small molecule inhibitors of the cMET receptor tyrosine kinase have been evaluated in clinical trials for the treatment of cancer and resistance-conferring mutations of cMET are beginning to be reported for a number of such compounds. There is now a need to understand specific cMET mutations at the molecular level, particularly concerning small molecule recognition. Toward this end, we report here the first crystal structures of the recent clinically observed resistance-conferring D1228V cMET mutant in complex with small molecule inhibitors, along with a crystal structure of wild-type cMET bound by the clinical compound savolitinib and supporting cellular, biochemical, and biophysical data. Our findings indicate that the D1228V alteration induces conformational changes in the kinase, which could have implications for small molecule inhibitor design. The data we report here increases our molecular understanding of the D1228V cMET mutation and provides insight for future inhibitor design.

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Conflict of interest statement

The authors declare the following competing financial interest(s): All authors are or were employees of AstraZeneca.

Figures

Figure 1
Figure 1
Type-I (crizotinib and savolitinib) and type-II (foretinib and BMS-777607) small molecule cMET inhibitors studied in this work.
Figure 2
Figure 2
Cellular and in vitro small molecule inhibition studies of wild-type and D1228V cMET. (a–b) Growth inhibition studies (three-day proliferation assay) of savolitinib (a) and foretinib (b) using parental NCI-H1993 cells (blue trace) and CRISPR/Cas9 modified NCI-H1993 cells expressing D1228V cMET (red trace). Values shown are average of three separate experiments. (c) Western blot analysis of NCI-H1993 and NCI-H1993 D1228V cMET cells treated with 1 μM savolitinib or foretinib for 4 h. (d–e) HTRF-based assessment of four small molecules in NCI-H1993 cells (d) and NCI-H1993 D1228V cMET cells (e). (f–g) Enzyme inhibition studies of four small molecules using wild-type (f) or D1228V (g) cMET. All potencies determined from these experiments except that determined for savolitinib in panel (g) are beyond the theoretical tight binding limit of the assay. Further experimental details can be found in Table S1 and in the Supporting Information.
Figure 3
Figure 3
Crystal structure of wild-type cMET bound by savolitinib, highlighting the key role that residues Y1230, K1110, and D1228 play in the inhibitor binding mode.
Figure 4
Figure 4
Crystal structures of D1228V cMET-inhibitor complexes. (a) Crystal structures of cMET bound by foretinib. Left panel: wild-type cMET bound by foretinib (PDB entry 3LQ8(30)). Right panel: D1228V cMET bound by foretinib, highlighting ordering of the A-loop (yellow). (b) Crystal structures of cMET bound by BMS-777607. Left panel: wild-type cMET bound by BMS-777607 (PDB entry 3F82(31)). Right panel: D1228V cMET bound by BMS-777607, highlighting ordering of the A-loop (yellow).
See this image and copyright information in PMC

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