Duff Michael R, Gabel Scott A, Pedersen Lars C, DeRose Eugene F, Krahn Juno M, Howell Elizabeth E, London Robert E
Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, Tennessee 37996, United States.
Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, 111 T. W. Alexander Drive, Research Triangle Park, Durham, North Carolina 27709, United States.
J Med Chem. 2020 Aug 13;63(15):8314-8324. doi: 10.1021/acs.jmedchem.0c00546. Epub 2020 Jul 28.
Although nonsteroidal anti-inflammatory drugs (NSAIDs) target primarily cyclooxygenase enzymes, a subset of NSAIDs containing carboxylate groups also has been reported to competitively inhibit dihydrofolate reductase (DHFR). In this study, we have characterized NSAID interactions with human DHFR based on kinetic, NMR, and X-ray crystallographic methods. The NSAIDs target a region of the folate binding site that interacts with the -aminobenzoyl-l-glutamate (pABG) moiety of folate and inhibit cooperatively with ligands that target the adjacent pteridine-recognition subsite. NSAIDs containing benzoate or salicylate groups were identified as having the highest potency. Among those tested, diflunisal, a salicylate derivative not previously identified to have anti-folate activity, was found to have a of 34 μM, well below peak plasma diflunisal levels reached at typical dosage levels. The potential of these drugs to interfere with the inflammatory process by multiple pathways introduces the possibility of further optimization to design dual-targeted analogs.
尽管非甾体抗炎药（NSAIDs）主要作用于环氧化酶，但据报道，一部分含有羧基的NSAIDs也能竞争性抑制二氢叶酸还原酶（DHFR）。在本研究中，我们基于动力学、核磁共振和X射线晶体学方法，对NSAIDs与人类DHFR的相互作用进行了表征。NSAIDs作用于叶酸结合位点的一个区域，该区域与叶酸的对氨基苯甲酰-L-谷氨酸（pABG）部分相互作用，并与作用于相邻蝶啶识别亚位点的配体协同抑制。含有苯甲酸酯或水杨酸酯基团的NSAIDs被确定具有最高的效力。在测试的药物中，双氯芬酸是一种先前未被鉴定具有抗叶酸活性的水杨酸酯衍生物，其抑制常数为34μM，远低于典型剂量水平下达到的血浆双氯芬酸峰值水平。这些药物通过多种途径干扰炎症过程的可能性为设计双靶点类似物的进一步优化提供了可能。
