Macromolecular Crystallography Laboratory, National Cancer Institute, Frederick, MD, USA.
Macromolecular Crystallography Laboratory, National Cancer Institute, Frederick, MD, USA; School of Chemistry and Chemical Engineering, Huaiyin Normal University, Huaiyin, Jiangsu Province, China(1).
Bioorg Med Chem. 2021 Jan 1;29:115847. doi: 10.1016/j.bmc.2020.115847. Epub 2020 Nov 9.
6-Hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK) is a key enzyme in the folate biosynthesis pathway. It catalyzes pyrophosphoryl transfer from ATP to 6-hydroxymethyl-7,8-dihydropterin (HP). HPPK is essential for microorganisms but absent in mammals; therefore, it is an attractive target for developing novel antimicrobial agents. Previously, based on our studies of the structure and mechanism of HPPK, we created first-generation bisubstrate inhibitors by linking 6-hydroxymethylpterin to adenosine through phosphate groups, and developed second-generation inhibitors by replacing the phosphate bridge with a linkage that contains a piperidine moiety. Here, we report third-generation inhibitors designed based on the piperidine-containing inhibitor, mimicking the transition state. We synthesized two such inhibitors, characterized their protein-binding and enzyme inhibition properties, and determined their crystal structures in complex with HPPK, advancing the development of such bisubstrate analog inhibitors.
6-羟甲基-7,8-二氢蝶呤磷酸化酶（HPPK）是叶酸生物合成途径中的关键酶。它催化从 ATP 到 6-羟甲基-7,8-二氢蝶呤（HP）的焦磷酸基转移。HPPK 对微生物是必需的，但哺乳动物中却不存在；因此，它是开发新型抗菌药物的有吸引力的靶标。先前，基于我们对 HPPK 结构和机制的研究，我们通过将 6-羟甲基喋呤通过磷酸基团连接到腺苷上来创建第一代双底物抑制剂，并通过用含有哌啶部分的键取代磷酸桥来开发第二代抑制剂。在这里，我们报告了基于含哌啶抑制剂设计的第三代抑制剂，模拟了过渡态。我们合成了两种这样的抑制剂，表征了它们的蛋白结合和酶抑制特性，并确定了它们与 HPPK 复合物的晶体结构，推进了这种双底物类似物抑制剂的开发。
