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Targeting enteropeptidase with reversible covalent inhibitors to achieve metabolic benefits

DOI: 10.1124/jpet.120.000219 DOI Help

Authors: Weimei Sun (Janssen Research & Development, LLC) , Xuqing Zhang (Janssen Research & Development, LLC) , Maxwell D. Cummings (Janssen Research & Development, LLC) , Kamal Albarazanji (Janssen Research & Development, LLC) , Jiejun Wu (Janssen Research & Development, LLC) , Mina Wang (Janssen Research & Development, LLC) , Richard Alexander (Janssen Research & Development, LLC) , Bin Zhu (Janssen Research & Development, LLC) , Yuemei Zhang (Janssen Research & Development, LLC) , James Leonard (Janssen Research & Development, LLC) , James Lanter (Janssen Research & Development, LLC) , James Lenhard (Janssen Research & Development, LLC)
Co-authored by industrial partner: Yes

Type: Journal Paper
Journal: Journal Of Pharmacology And Experimental Therapeutics

State: Published (Approved)
Published: October 2020

Abstract: Inhibition of the serine protease enteropeptidase (EP) opens a new avenue to the discovery of chemotherapeutics for the treatment of metabolic diseases. Camostat has been used clinically for treating chronic pancreatitis in Japan; however, the mechanistic basis of the observed clinical efficacy has not been fully elucidated. We demonstrate that camostat is a potent reversible covalent inhibitor of EP, with an inhibition potency (kinact/KI) of 1.5 x 104 M-1s-1. High-resolution LC-MS showed addition of 161.6 Da to EP following reaction with camostat, consistent with insertion of the carboxyphenylguanidine moiety of camostat. Covalent inhibition of EP by camostat is reversible, with an enzyme reactivation half-life of 14.3 hours. Formation of a covalent adduct was further supported by a crystal structure resolved to 2.19Å, showing modification of the catalytic serine of EP by a close analog of camostat leading to formation of the carboxyphenylguanidine acyl enzyme identical to that expected for reaction with camostat. Of particular note, minor structural modifications of camostat led to changes in the mechanism of inhibition. We observed from other studies that sustained inhibition of EP is required to effect a reduction in cumulative food intake and body weight, with concomitant improved blood glucose levels in obese and diabetic ob/ob mice. Thus, the structure-activity relationship (SAR) needs to be driven by not only the inhibition potency but also the mechanistic and kinetic characterization. Our findings support EP as a target for the treatment of metabolic diseases, and demonstrate that reversible covalent EP inhibitors show clinically relevant efficacy.

Journal Keywords: covalent drug binding; inhibition; mechanism-based inhibition; protease inhibitors

Subject Areas: Biology and Bio-materials, Medicine

Instruments: I02-Macromolecular Crystallography