Structure-based design of submicromolar, biologically active inhibitors of trypanosomatid glyceraldehyde-3-phosphate dehydrogenase
Structure-based design of submicromolar, biologically active inhibitors of trypanosomatid glyceraldehyde-3-phosphate dehydrogenase
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DOI：
10.1073/pnas.96.8.4273
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发表时间：
1999-04-13
影响因子：
11.1
通讯作者：
Gelb, MH
中科院分区：
文献类型：
--
作者：
Aronov, AM;Suresh, S;Gelb, MH
布氏锥虫的血流阶段和克氏锥虫的胞内(无鞭毛体)阶段的能量全部来自糖酵解，抑制糖酵解酶可能是开发抗锥虫药物的一种新途径，前提是获得足够的寄生虫对宿主的选择性。根据人、布氏毛滴虫和墨西哥利什曼原虫甘油醛-3-磷酸脱氢酶的晶体结构，我们设计了腺苷类似物作为紧密结合的抑制剂，占据了NAD(+)共底物的腺苷部分的口袋。虽然腺苷是一种很差的抑制剂，IC50接近50 mM，但在核糖的2‘位和腺苷的N-6位上添加取代基，导致在甘油醛-3-磷酸脱氢酶测定中的二取代核苷具有微摩尔到亚微摩尔的效力，比铅提高了5个数量级。所设计的化合物在测试达到其溶解度极限(约40微米)时不会抑制人类糖酵解酶。当对培养的布鲁氏毛滴虫和细胞内T，CRUZI进行测试时，N-6-(1-naphthalenemethyl)-2‘-(3-chlorobenzamido)adenosine在低微摩尔范围内抑制生长。在将该化合物加入血液T、布鲁斯后的几分钟内，葡萄糖衍生的丙酮酸的产生停止，寄生虫的运动丧失，并观察到严重变形和溶解的寄生虫的混合物。这些研究强调了使用基于结构的药物设计将平庸的先导化合物转化为有效的酶抑制剂的可行性。他们还表明，锥虫的能量生产可以通过糖酵解途径中一种酶的紧密结合的竞争性抑制物来阻止。
The bloodstream stage of Trypanosoma brucei and probably the intracellular (amastigote) stage of Trypanosoma cruzi derive all of their energy from glycolysis, Inhibiting glycolytic enzymes may be a novel approach for the development of antitrypanosomatid drugs provided that sufficient parasite versus host selectivity can be obtained. Guided by the crystal structures of human, T. brucei, and Leishmania mexicana glyceraldehyde-3-phosphate dehydrogenase, we designed adenosine analogs as tight binding inhibitors that occupy the pocket on the enzyme that accommodates the adenosyl moiety of the NAD(+) cosubstrate. Although adenosine is a very poor inhibitor, IC50 approximate to 50 mM, addition of substituents to the 2' position of ribose and the N-6-position of adenosine led to disubstituted nucleosides with micromolar to submicromolar potency in glyceraldehyde-3-phosphate dehydrogenase assays, an improvement of 5 orders of magnitude over the lead. The designed compounds do not inhibit the human glycolytic enzyme when tested up to their solubility limit (approximate to 40 mu M). When tested against cultured bloodstream T. brucei and intracellular T, cruzi, N-6-(1-naphthalenemethyl)-2'-(3-chlorobenzamido)adenosine inhibited growth in the low micromolar range. Within minutes after adding this compound to bloodstream T, brucei, production of glucose-derived pyruvate ceased, parasite motility was lost, and a mixture of grossly deformed and lysed parasites was observed. These studies underscore the feasibility of using structure-based drug design to transform a mediocre lead compound into a potent enzyme inhibitor. They also suggest that energy production can be blocked in trypanosomatids with a tight binding competitive inhibitor of an enzyme in the glycolytic pathway.