Fragment Growing and Linking Lead to Novel Nanomolar Lactate Dehydrogenase Inhibitors
Fragment Growing and Linking Lead to Novel Nanomolar Lactate Dehydrogenase Inhibitors
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DOI：
10.1021/jm3014844
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发表时间：
2013-02-14
影响因子：
7.3
通讯作者：
Dalgarno, David C.
中科院分区：
文献类型：
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作者：
Kohlmann, Anna;Zech, Stephan G.;Dalgarno, David C.
乳酸脱氢酶A(LDH-A)催化糖酵解途径中乳酸和丙酮酸的相互转化。癌细胞严重依赖糖酵解而不是氧化磷酸化来产生三磷酸腺苷，这种现象被称为华宝效应。因此，小分子对LDH-A的抑制对于潜在的癌症治疗是有意义的。我们描述了通过基于片段的药物发现来鉴定和优化LDH-A抑制剂。我们应用基于配基的核磁共振筛选来鉴定与LDH-A结合的低亲和力片段。用表面等离子体共振法(SPR)和酶分析法分别测定碎片HITS的解离常数(K-d)和酶抑制(IC50)。用X射线结晶学研究了所选片段的结合方式。片段生长和连接，然后进行化学优化，得到了纳米分子LDH-A抑制剂，证明了与LDH-A的化学计量结合。选定的分子抑制细胞中乳酸的产生，表明对癌细胞系具有靶向性抑制作用。
Lactate dehydrogenase A (LDH-A) catalyzes the inter-conversion of lactate and pyruvate in the glycolysis pathway. Cancer cells rely heavily on glycolysis instead of oxidative phosphorylation to generate ATP, a phenomenon known as the Warburg effect. The inhibition of LDH-A by small molecules is therefore of interest for potential cancer treatments. We describe the identification and optimization of LDH-A inhibitors by fragment-based drug discovery. We applied ligand based NMR screening to identify low affinity fragments binding to LDH-A. The dissociation constants (K-d) and enzyme inhibition (IC50) of fragment hits were measured by surface plasmon resonance (SPR) and enzyme assays, respectively. The binding modes of selected fragments were investigated by X-ray crystallography. Fragment growing and linking, followed by chemical optimization, resulted in nanomolar LDH-A inhibitors that demonstrated stoichiometric binding to LDH-A. Selected molecules inhibited lactate production in cells, suggesting target-specific inhibition in cancer cell lines.