Discovery of novel allosteric site and covalent inhibitors of FBPase with potent hypoglycemic effects
Discovery of novel allosteric site and covalent inhibitors of FBPase with potent hypoglycemic effects
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发现具有有效降血糖作用的新型变构位点和 FBPase 共价抑制剂
DOI：
10.1016/j.ejmech.2019.111749
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发表时间：
2019
影响因子：
6.7
通讯作者：
Wan Jian
中科院分区：
文献类型：
--
作者：
Huang Yunyuan;Wei Lin;Han Xinya;Chen Haifeng;Ren Yanliang;Xu Yanhong;Song Rongrong;Rao Li;Su Chen;Peng Chao;Feng Lingling;Wan Jian
1，6-二磷酸果糖酶(FBPase)是GNG途径中的一个重要酶。显著的进展表明，FBPase在糖尿病的治疗中发挥着关键作用。针对AMP位点开发了许多FBPase抑制剂，但这些抑制剂都没有表现出合适的药效和可药性。在此，我们发现了FBPase上的一个新的变构位点(C128)，并发现了几种对FBPase具有强烈抑制作用的硝基苯乙烯化合物与FBPase上的C128位点发生了共价结合。突变研究表明，C128是唯一能影响FBPase抑制的半胱氨酸，N125-S124-S123途径很可能参与了C128与活性部位之间的变构信号传递。然而，这些硝基苯乙烯除了与FBPase中的C128结合外，还可能与多个半胱氨酸结合。为了提高口袋选择性，通过整合基于片段的共价虚拟筛选和基于机器学习的综合复杂性评估，对一系列新的化合物(14a-14n)进行了合理的重新设计。正如预期的那样，质谱学验证了FBPase中与C128结合的标题化合物的比例显著高于硝基苯乙烯。值得注意的是，在生理和病理条件下，化合物14b、14c、14i或14n的处理显著抑制了葡萄糖的产生，并降低了小鼠原代肝细胞的甘油三酯和总胆固醇水平。我们强调了一种新的范例，即分子靶向FBPase上的C128位点可以具有强大的降血糖作用。
Fructose-1,6-bisphosphatase (FBPase) is an essential enzyme of GNG pathway. Significant advances demonstrate the FBPase plays a critical role in treatment of diabetes. Numerous FBPase inhibitors were developed by targeting AMP site, nevertheless, none of these inhibitors has exhibited suitable potency and druggability. Herein, a new allosteric site (C128) on FBPase was discovered, and several nitrostyrene compounds exhibiting potent FBPase inhibitions were found covalently bind to C128 site on FBPase. Mutagenesis suggest that C128 is the only cysteine that can influence FBPase inhibition, the N125–S124–S123 pathway was most likely involved in allosteric signaling transmission between C128 and active site. However, these nitrostyrenes may bind with multiple cysteine besides C128 in FBPase. To improve pocket selectivity, a series of novel compounds (14a-14n)were re-designed rationally by integrating fragment-based covalent virtual screening and machine-learning-based synthetic complexity evaluation. As expected, the mass spectrometry validated that the proportion of title compounds binding to the C128 in FBPase was significantly higher than that of nitrostyrenes. Notably, under physiological and pathological conditions, the treatment of compounds14b, 14c, 14ior14nled to potent inhibition of glucose production, as well as decreased triglyceride and total cholesterol levels in mouse primary hepatocytes. We highlight a novel paradigm that molecular targeting C128 site on FBPase can have potent hypoglycemic effect.