Nunes Jéssica Alves, Santos-Júnior Paulo Fernando da Silva, Gomes Midiane Correa, Ferreira Luiz Alberto Santos, Padilha Emanuelly Karla Araújo, Teixeira Thaiz Rodrigues, Stanger Emily J, Kaur Yashpreet, Silva Elany Barbosa da, Costa Clara Andrezza Crisóstomo Bezerra, Freitas Johnnatan Duarte de, Araújo-Júnior João Xavier de, Mendonça-Junior Francisco Jaime Bezerra, Giardini Miriam A, Siqueira-Neto Jair L, Caffrey Conor R, Zhan Peng, Cardoso Sílvia Helena, Silva-Júnior Edeildo Ferreira da
Biological and Molecular Chemistry Research Group, Institute of Chemistry and Biotechnology, Federal University of Alagoas, Lourival Melo Mota Avenue, AC. Simões Campus, Alagoas, Maceió, 57072-970, Brazil.
Laboratory of Medicinal Chemistry, Institute of Pharmaceutical Sciences, Federal University of Alagoas, Lourival Melo Mota Avenue, AC. Simões Campus, Alagoas, Maceió, 57072-970, Brazil.
Eur J Med Chem. 2025 Feb 5;283:117109. doi: 10.1016/j.ejmech.2024.117109. Epub 2024 Nov 28.
Trypanosoma cruzi (T. cruzi) and Trypanosoma brucei (T. brucei) urgently demand innovative drug development due to their impact on public health worldwide. Their cysteine proteases, Cruzain (CRZ) and the T. brucei Cathepsin L-like protease (TbrCATL) are established drug targets for these parasites. In this study, our coumarin-3-thiosemicarbazones demonstrated nanomolar IC values (22-698 nM) toward these proteases. Against T. cruzi amastigotes and T. brucei trypomastigotes, LASF-01 displayed a promising result. Herein, MCG-02, the most potent TbrCATL inhibitor, underwent comprehensive analyses, including cytotoxicity assessments and in vitro tests. Molecular dynamics (MD) simulations and a multiscale Quantum Mechanics/Quantum Mechanics (QM/QM) approach were used to generate insights into their binding modes. These suggested that MCG-02 could be a reversible, competitive covalent inhibitor. Further, confirmatory assays were experimentally performed changing different parameters to prove its efficacy. Additionally, the predicted pharmacokinetic profile showed that there is no violation of the Lipinski rule of five. Notably, coumarin-3-thiosemicarbazone hybrids emerged as promising candidates for designing highly active inhibitors against CRZ and TbrCATL. Overall, the integration of in silico and experimental approaches enhanced our understanding regarding the binding modes of MCG-02, which were experimentally corroborated, providing valuable insights for future drug development.
克氏锥虫（T. cruzi）和布氏锥虫（T. brucei）因其对全球公共卫生的影响，迫切需要创新药物研发。它们的半胱氨酸蛋白酶，即克氏锥虫蛋白酶（CRZ）和布氏锥虫组织蛋白酶L样蛋白酶（TbrCATL），是这些寄生虫已确定的药物靶点。在本研究中，我们的香豆素-3-硫代氨基脲对这些蛋白酶表现出纳摩尔级的IC值（22 - 698 nM）。针对克氏锥虫无鞭毛体和布氏锥虫锥鞭毛体，LASF - 01显示出有前景的结果。在此，最有效的TbrCATL抑制剂MCG - 02进行了全面分析，包括细胞毒性评估和体外试验。利用分子动力学（MD）模拟和多尺度量子力学/量子力学（QM/QM）方法深入了解它们的结合模式。这些结果表明MCG - 02可能是一种可逆的、竞争性共价抑制剂。此外，通过改变不同参数进行了实验性的验证试验以证明其有效性。另外，预测的药代动力学概况表明没有违反Lipinski五规则。值得注意的是，香豆素-3-硫代氨基脲杂化物成为设计针对CRZ和TbrCATL的高活性抑制剂的有前景的候选物。总体而言，计算机模拟和实验方法的结合增强了我们对MCG - 02结合模式的理解，这在实验中得到了证实，为未来的药物研发提供了有价值的见解。
