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Prolonged and tunable residence time using reversible covalent kinase inhibitors

Abstract

Drugs with prolonged on-target residence times often show superior efficacy, yet general strategies for optimizing drug-target residence time are lacking. Here we made progress toward this elusive goal by targeting a noncatalytic cysteine in Bruton's tyrosine kinase (BTK) with reversible covalent inhibitors. Using an inverted orientation of the cysteine-reactive cyanoacrylamide electrophile, we identified potent and selective BTK inhibitors that demonstrated biochemical residence times spanning from minutes to 7 d. An inverted cyanoacrylamide with prolonged residence time in vivo remained bound to BTK for more than 18 h after clearance from the circulation. The inverted cyanoacrylamide strategy was further used to discover fibroblast growth factor receptor (FGFR) kinase inhibitors with residence times of several days, demonstrating the generalizability of the approach. Targeting of noncatalytic cysteines with inverted cyanoacrylamides may serve as a broadly applicable platform that facilitates 'residence time by design', the ability to modulate and improve the duration of target engagement in vivo.

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Figure 1: Reversible covalent BTK inhibitors based on inverted cyanoacrylamides.
Figure 2: Prolonged and tunable residence time of reversible covalent BTK inhibitors.
Figure 3: Long-term cellular durability of reversible covalent BTK inhibitors.
Figure 4: Kinase selectivity of inhibitor 9.
Figure 5: Extended pharmacodynamic effect of an orally bioavailable BTK inhibitor.
Figure 6: Inverted cyanoacrylamide FGFR inhibitors with prolonged, tunable residence times.

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Acknowledgements

This work was supported by the US National Institutes of Health (NIH) (grants GM071434 to J.T. and F32GM087052 to J.M.M.), the University of California San Francisco (UCSF) Stephen & Nancy Grand Multiple Myeloma Translational Initiative (to J.T.), the Academy of Finland (to V.O.P.) and the Sigrid Juselius Foundation (to V.O.P.). We acknowledge the UCSF Mass Spectrometry Facility (supported by NIH grant P41RR001614).

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J.M.M., V.O.P. and J.T. designed experiments involving compounds 13; J.M.M. and V.O.P. performed experiments and analyzed data for those compounds. J.M.B., A.B., D.T., V.T.P., S.R., P.A.N., D.E.K., M.E.G., J.O.F., T.D.O., E.V., K.A.B., R.J.H. and D.M.G. designed and managed experiments involving compounds 446; J.M.B., A.B., D.T., V.T.P., S.R., D.F., J.S., V.P., T.T., X.L. and D.G.L. performed experiments and analyzed data for those compounds. J.M.B. and J.T. wrote the manuscript with feedback from other authors, and all authors read and approved the manuscript.

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Correspondence to J Michael Bradshaw or Jack Taunton.

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Competing interests

J.T., J.M.M. and V.O.P. have filed patent applications on cyanoacrylamide kinase inhibitors (licensed to Principia Biopharma, of which J.T. is a cofounder). J.M.B., A.B., D.T., V.T.P., D.F., J.S., V.P., T.T., X.L., D.G.L., P.A.N., D.E.K., M.E.G., J.O.F., T.D.O., E.V., K.A.B., R.J.H. and D.M.G. are members of Principia Biopharma, which is interested in developing BTK inhibitors for therapeutic applications.

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Supplementary Results, Supplementary Figures 1–25, Supplementary Tables 1–6 and Supplementary Note. (PDF 4153 kb)

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Bradshaw, J., McFarland, J., Paavilainen, V. et al. Prolonged and tunable residence time using reversible covalent kinase inhibitors. Nat Chem Biol 11, 525–531 (2015). https://doi.org/10.1038/nchembio.1817

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