1. Academic Validation
  2. Dual Allosteric Inhibition of SHP2 Phosphatase

Dual Allosteric Inhibition of SHP2 Phosphatase

  • ACS Chem Biol. 2018 Mar 16;13(3):647-656. doi: 10.1021/acschembio.7b00980.
Michelle Fodor 1 Edmund Price 1 Ping Wang 1 Hengyu Lu 1 Andreea Argintaru 1 Zhouliang Chen 1 Meir Glick 1 Huai-Xiang Hao 1 Mitsunori Kato 1 Robert Koenig 1 Jonathan R LaRochelle 2 Gang Liu 1 Eric McNeill 1 Dyuti Majumdar 1 Gisele A Nishiguchi 1 Lawrence B Perez 1 Gregory Paris 1 Christopher M Quinn 1 Timothy Ramsey 1 Martin Sendzik 1 Michael David Shultz 1 Sarah L Williams 1 Travis Stams 1 Stephen C Blacklow 2 Michael G Acker 1 Matthew J LaMarche 1
Affiliations

Affiliations

  • 1 Novartis Institutes for Biomedical Research , Cambridge , Massachusetts 02139 , United States.
  • 2 Department of Biological Chemistry & Molecular Pharmacology , Harvard Medical School and Department of Cancer Biology, Dana-Farber Cancer Institute , Boston , Massachusetts 02215 , United States.
Abstract

SHP2 is a cytoplasmic protein tyrosine Phosphatase encoded by the PTPN11 gene and is involved in cell proliferation, differentiation, and survival. Recently, we reported an allosteric mechanism of inhibition that stabilizes the auto-inhibited conformation of SHP2. SHP099 (1) was identified and characterized as a moderately potent, orally bioavailable, allosteric small molecule inhibitor, which binds to a tunnel-like pocket formed by the confluence of three domains of SHP2. In this report, we describe further screening strategies that enabled the identification of a second, distinct small molecule allosteric site. SHP244 (2) was identified as a weak inhibitor of SHP2 with modest thermal stabilization of the Enzyme. X-ray crystallography revealed that 2 binds and stabilizes the inactive, closed conformation of SHP2, at a distinct, previously unexplored binding site-a cleft formed at the interface of the N-terminal SH2 and PTP domains. Derivatization of 2 using structure-based design resulted in an increase in SHP2 thermal stabilization, biochemical inhibition, and subsequent MAPK pathway modulation. Downregulation of DUSP6 mRNA, a downstream MAPK pathway marker, was observed in KYSE-520 Cancer cells. Remarkably, simultaneous occupation of both allosteric sites by 1 and 2 was possible, as characterized by cooperative biochemical inhibition experiments and X-ray crystallography. Combining an allosteric site 1 inhibitor with an allosteric site 2 inhibitor led to enhanced pharmacological pathway inhibition in cells. This work illustrates a rare example of dual allosteric targeted protein inhibition, demonstrates screening methodology and tactics to identify allosteric inhibitors, and enables further interrogation of SHP2 in Cancer and related pathologies.

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