Novel mutant-selective EGFR kinase inhibitors against EGFR T790M


The clinical efficacy of epidermal growth factor receptor (EGFR) kinase inhibitors in EGFR-mutant non-small-cell lung cancer (NSCLC) is limited by the development of drug-resistance mutations, including the gatekeeper T790M mutation1,2,3. Strategies targeting EGFR T790M with irreversible inhibitors have had limited success and are associated with toxicity due to concurrent inhibition of wild-type EGFR4,5. All current EGFR inhibitors possess a structurally related quinazoline-based core scaffold and were identified as ATP-competitive inhibitors of wild-type EGFR. Here we identify a covalent pyrimidine EGFR inhibitor by screening an irreversible kinase inhibitor library specifically against EGFR T790M. These agents are 30- to 100-fold more potent against EGFR T790M, and up to 100-fold less potent against wild-type EGFR, than quinazoline-based EGFR inhibitors in vitro. They are also effective in murine models of lung cancer driven by EGFR T790M. Co-crystallization studies reveal a structural basis for the increased potency and mutant selectivity of these agents. These mutant-selective irreversible EGFR kinase inhibitors may be clinically more effective and better tolerated than quinazoline-based inhibitors. Our findings demonstrate that functional pharmacological screens against clinically important mutant kinases represent a powerful strategy to identify new classes of mutant-selective kinase inhibitors.

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Figure 1: WZ4002, WZ3146 and WZ8040 are novel EGFR inhibitors, suppress the growth of EGFR -T790M-containing cell lines and inhibit EGFR phosphorylation.
Figure 2: WZ4002 is less potent than quinazoline EGFR inhibitors against WT EGFR in vitro and in vivo.
Figure 3: Crystal structure of WZ4002 bound to EGFR T790M.
Figure 4: WZ4002 inhibits EGFR phosphorylation and induces significant tumour regression in murine models of EGFR T790M.

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Protein Data Bank

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Structural data are deposited in Protein Data Bank under accession number 3IKA.


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This study is supported by grants from the National Institutes of Health RO1CA11446 (P.A.J.), R01CA135257 (P.A.J.), R01CA080942 (M.J.E.), R01CA130876-02 (N.S.G.), R01CA116020 (M.J.E.), R01AG2400401 (K.-K.W.), R01 CA122794 (K.-K.W.), R01GM070590 (J.R.E.), National Cancer Institute Lung SPORE P50CA090578 (P.A.J. and K.-K.W.), the Cecily and Robert Harris Foundation (K.-K.W.), Uniting Against Lung Cancer (K.-K.W.), the Flight Attendant Medical Research Institute (K.-K.W.)., the Hazel and Samuel Bellin Research Fund (P.A.J.) and the Damon Runyon Foundation Cancer Innovation Award (N.S.G.). This work is contribution 948 from the Barnett Institute. The authors thank Sai Advantium Pharma for performing the pharmacokinetic studies.

Author Contributions W.Z., D.E., Li.C., C.-H.Y., D.L., M.C. A.B.C. designed experiments, conducted studies and analysed data. R.E.I. and J.R.E. performed and analysed the mass spectrometry studies. Lu.C. and R.P. performed the histological and immunohistochemistry analyses. M.J.E., K.-K.W., N.S.G. and P.A.J. designed the experiments, analysed data and wrote the manuscript. The Wong, Eck, Gray and Jänne laboratories contributed equally to this work.

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Correspondence to Nathanael S. Gray or Pasi A. Jänne.

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Competing interests

P.A.J. receives royalties as a co-inventor on a patent awarded for the discovery of EGFR mutations, licensed to Genzyme Genetics, which was not involved in this study. M.J.E. is a consultant for and receives research support from Novartis Institutes for Biomedical Research. N.S.G., M.J.E. and P.A.J. are co-inventors on a provisional patent application covering the inhibitors described in the manuscript which have been licensed to Gatekeeper Pharmaceuticals of which N.S.G., K.-K.W. and P.A.J. are co-founders.

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This file contains Supplementary Methods, Supplementary Tables S1- S10, Supplementary Figures S1-S11 with Legends and Supplementary References. (PDF 1228 kb)

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Zhou, W., Ercan, D., Chen, L. et al. Novel mutant-selective EGFR kinase inhibitors against EGFR T790M. Nature 462, 1070–1074 (2009).

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