Hoarau Marie, Vanichtanankul Jarunee, Srimongkolpithak Nitipol, Vitsupakorn Danoo, Yuthavong Yongyuth, Kamchonwongpaisan Sumalee
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathumthani, Thailand.
J Enzyme Inhib Med Chem. 2021 Dec;36(1):198-206. doi: 10.1080/14756366.2020.1854244.
In various malaria-endemic regions, the appearance of resistance has precluded the use of pyrimidine-based antifolate drugs. Here, a three-step fragment screening was used to identify new non-pyrimidine dihydrofolate reductase (DHFR) inhibitors. Starting from a 1163-fragment commercial library, a two-step differential scanning fluorimetry screen identified 75 primary fragment hits. Subsequent enzyme inhibition assay identified 11 fragments displaying IC in the 28-695 μM range and selectivity for DHFR. In addition to the known pyrimidine, three new anti-DHFR chemotypes were identified. Fragments from each chemotype were successfully co-crystallized with DHFR, revealing a binding in the active site, in the vicinity of catalytic residues, which was confirmed by molecular docking on all fragment hits. Finally, comparison with similar non-hit fragments provides preliminary input on available growth vectors for future drug development.
在各个疟疾流行地区，耐药性的出现使得基于嘧啶的抗叶酸药物无法使用。在此，采用三步片段筛选法来鉴定新型非嘧啶二氢叶酸还原酶（DHFR）抑制剂。从一个包含1163个片段的商业文库开始，两步差示扫描荧光法筛选鉴定出75个初步片段命中物。随后的酶抑制试验鉴定出11个片段，其IC值在28 - 695μM范围内，且对DHFR具有选择性。除了已知的嘧啶外，还鉴定出三种新的抗DHFR化学类型。来自每种化学类型的片段成功地与DHFR共结晶，揭示了其在活性位点、催化残基附近的结合情况，这通过对所有片段命中物的分子对接得到了证实。最后，与类似的未命中片段进行比较，为未来药物开发的可用生长载体提供了初步信息。
