Department of Biochemistry & Molecular Biophysics, Washington University School of Medicine, Saint Louis, Missouri 63110, United States.
Protein Structure Laboratory, Del Shankel Structural Biology Center, University of Kansas, Lawrence, Kansas 66047, United States.
J Med Chem. 2024 Mar 28;67(6):4833-4854. doi: 10.1021/acs.jmedchem.3c02388. Epub 2024 Mar 13.
Protease inhibitor drug discovery is challenged by the lack of cellular and oral permeability, selectivity, metabolic stability, and rapid clearance of peptides. Here, we describe the rational design, synthesis, and evaluation of peptidomimetic side-chain-cyclized macrocycles which we converted into covalent serine protease inhibitors with the addition of an electrophilic ketone warhead. We have identified potent and selective inhibitors of TMPRSS2, matriptase, hepsin, and HGFA and demonstrated their improved protease selectivity, metabolic stability, and pharmacokinetic (PK) properties. We obtained an X-ray crystal structure of phenyl ether-cyclized tripeptide VD4162 () bound to matriptase, revealing an unexpected binding conformation. Cyclic biphenyl ether VD5123 () displayed the best PK properties in mice with a half-life of 4.5 h and compound exposure beyond 24 h. These new cyclic tripeptide scaffolds can be used as easily modifiable templates providing a new strategy to overcoming the obstacles presented by linear acyclic peptides in protease inhibitor drug discovery.
蛋白酶抑制剂的药物研发面临着细胞和口服通透性差、选择性差、代谢稳定性差、肽类快速清除等问题。在这里，我们描述了具有合理设计、合成和评价的肽模拟侧链环化大环，我们通过添加亲电酮弹头将其转化为共价丝氨酸蛋白酶抑制剂。我们已经鉴定出 TMPRSS2、matriptase、hepsin 和 HGFA 的有效且选择性抑制剂，并证明了它们改善的蛋白酶选择性、代谢稳定性和药代动力学（PK）特性。我们获得了苯醚环化三肽 VD4162（）与 matriptase 结合的 X 射线晶体结构，揭示了一种意想不到的结合构象。联苯醚 VD5123（）在小鼠中表现出最佳的 PK 特性，半衰期为 4.5 小时，化合物暴露时间超过 24 小时。这些新的环三肽支架可以用作易于修饰的模板，为克服蛋白酶抑制剂药物研发中线性非环肽所带来的障碍提供了一种新的策略。
