Abt Evan R, Rosser Ethan W, Durst Matthew A, Lok Vincent, Poddar Soumya, Le Thuc M, Cho Arthur, Kim Woosuk, Wei Liu, Song Janet, Capri Joseph R, Xu Shili, Wu Nanping, Slavik Roger, Jung Michael E, Damoiseaux Robert, Czernin Johannes, Donahue Timothy R, Lavie Arnon, Radu Caius G
Department of Molecular and Medical Pharmacology, University of California Los Angeles, Los Angeles, CA, USA; Ahmanson Translational Imaging Division, University of California Los Angeles, Los Angeles, CA, USA.
Ahmanson Translational Imaging Division, University of California Los Angeles, Los Angeles, CA, USA; Department of Chemistry and Biochemistry, University of California Los Angeles, Los Angeles, CA, USA.
Cell Chem Biol. 2020 Feb 20;27(2):197-205.e6. doi: 10.1016/j.chembiol.2019.10.012. Epub 2019 Nov 13.
Biosynthesis of the pyrimidine nucleotide uridine monophosphate (UMP) is essential for cell proliferation and is achieved by the activity of convergent de novo and salvage metabolic pathways. Here we report the development and application of a cell-based metabolic modifier screening platform that leverages the redundancy in pyrimidine metabolism for the discovery of selective UMP biosynthesis modulators. In evaluating a library of protein kinase inhibitors, we identified multiple compounds that possess nucleotide metabolism modifying activity. The JNK inhibitor JNK-IN-8 was found to potently inhibit nucleoside transport and engage ENT1. The PDK1 inhibitor OSU-03012 (also known as AR-12) and the RAF inhibitor TAK-632 were shown to inhibit the therapeutically relevant de novo pathway enzyme DHODH and their affinities were unambiguously confirmed through in vitro assays and co-crystallization with human DHODH.
嘧啶核苷酸尿苷一磷酸（UMP）的生物合成对细胞增殖至关重要，它通过从头合成和补救代谢途径的协同作用来实现。在此，我们报告了一种基于细胞的代谢调节剂筛选平台的开发与应用，该平台利用嘧啶代谢的冗余性来发现选择性UMP生物合成调节剂。在评估蛋白激酶抑制剂文库时，我们鉴定出了多种具有核苷酸代谢修饰活性的化合物。发现JNK抑制剂JNK-IN-8能有效抑制核苷转运并作用于ENT1。PDK1抑制剂OSU-03012（也称为AR-12）和RAF抑制剂TAK-632被证明可抑制具有治疗相关性的从头合成途径酶DHODH，并且通过体外试验以及与人DHODH的共结晶明确证实了它们的亲和力。
