Use of protease substrate specificity screening in the rational design of selective protease inhibitors with unnatural amino acids: Application to HGFA, matriptase, and hepsin.

Autor: Mahoney MW; Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, Saint Louis, Missouri, USA., Helander J; Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, Saint Louis, Missouri, USA., Kooner AS; Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, Saint Louis, Missouri, USA., Norman M; Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, Saint Louis, Missouri, USA., Damalanka VC; Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, Saint Louis, Missouri, USA., De Bona P; Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, Saint Louis, Missouri, USA., Kasperkiewicz P; Division of Chemical Biology and Bioimaging, Department of Chemistry, Wroclaw University of Science and Technology, Wroclaw, Poland., Rut W; Division of Chemical Biology and Bioimaging, Department of Chemistry, Wroclaw University of Science and Technology, Wroclaw, Poland., Poreba M; Division of Chemical Biology and Bioimaging, Department of Chemistry, Wroclaw University of Science and Technology, Wroclaw, Poland., Kashipathy MM; Protein Structure Laboratory, Del Shankel Structural Biology Center, University of Kansas, Lawrence, Kansas, USA., Battaile KP; New York Structural Biology Center, Upton, New York, USA., Lovell S; Protein Structure Laboratory, Del Shankel Structural Biology Center, University of Kansas, Lawrence, Kansas, USA., O'Donoghue AJ; Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, California, USA., Craik CS; Department of Pharmaceutical Chemistry, University of California, San Francisco, California, USA., Drag M; Division of Chemical Biology and Bioimaging, Department of Chemistry, Wroclaw University of Science and Technology, Wroclaw, Poland., Janetka JW; Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, Saint Louis, Missouri, USA.
Jazyk: angličtina
Zdroj: Protein science : a publication of the Protein Society [Protein Sci] 2024 Aug; Vol. 33 (8), pp. e5110.
DOI: 10.1002/pro.5110
Abstrakt: Inhibition of the proteolytic processing of hepatocyte growth factor (HGF) and macrophage stimulating protein (MSP) is an attractive approach for the drug discovery of novel anticancer therapeutics which prevent tumor progression and metastasis. Here, we utilized an improved and expanded version of positional scanning of substrate combinatorial libraries (PS-SCL) technique called HyCoSuL to optimize peptidomimetic inhibitors of the HGF/MSP activating serine proteases, HGFA, matriptase, and hepsin. These inhibitors have an electrophilic ketone serine trapping warhead and thus form a reversible covalent bond to the protease. We demonstrate that by varying the P2, P3, and P4 positions of the inhibitor with unnatural amino acids based on the protease substrate preferences learned from HyCoSuL, we can predictably modify the potency and selectivity of the inhibitor. We identified the tetrapeptide JH-1144 (8) as a single digit nM inhibitor of HGFA, matriptase and hepsin with excellent selectivity over Factor Xa and thrombin. These unnatural peptides have increased metabolic stability relative to natural peptides of similar structure. The tripeptide inhibitor PK-1-89 (2) has excellent pharmacokinetics in mice with good compound exposure out to 24 h. In addition, we obtained an X-ray structure of the inhibitor MM1132 (15) bound to matriptase revealing an interesting binding conformation useful for future inhibitor design.
(© 2024 The Protein Society.)
Databáze: MEDLINE
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