期刊：
JOURNAL OF MEDICINAL CHEMISTRY,2020年63(11):6238-6247 ISSN：0022-2623
通讯作者：
Ren, Yanliang;Wan, Jian;Li, Jian
作者机构：
[Feng, Lingling; Rao, Li; Wei, Lin; Ren, Yanliang; Wan, Jian; Xu, Yanhong; Liu, Jiaqi; Song, Rongrong; Huang, Yunyuan] Cent China Normal Univ, Coll Chem, Minist Educ, Key Lab Pesticide & Chem Biol CCNU, Wuhan 430079, Peoples R China.;[Xu, Yixiang; Li, Jian] East China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab New Drug Design, Shanghai 200237, Peoples R China.;[Ni, Shuaishuai] Shanghai Univ Tradit Chinese Med, Canc Inst, Longhua Hosp, Shanghai 200237, Peoples R China.;[Wang, Yingjie] Shenzhen Bay Lab, Shenzhen 518055, Guangdong, Peoples R China.;[Su, Chen; Peng, Chao] Natl Facil Prot Sci Shanghai, Zhangjiang Lab, Shanghai 201210, Peoples R China.
通讯机构：
[Ren, YL; Wan, J] C;[Li, Jian] E;Cent China Normal Univ, Coll Chem, Minist Educ, Key Lab Pesticide & Chem Biol CCNU, Wuhan 430079, Peoples R China.;East China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab New Drug Design, Shanghai 200237, Peoples R China.
摘要：
Fructose 1,6-bisphosphatase (FBPase) has attracted substantial interest as a target associated with cancer and type 2 diabetes. Herein, we found that disulfiram and its derivatives can potently inhibit FBPase by covalently binding to a new C128 allosteric site distinct from the original C128 site in APO FBPase. Further identification of the allosteric inhibition mechanism reveals that the covalent binding of a fragment of 214 will result in the movement of C128 and the dissociation of helix H4 (123-128), which in turn allows S123 to more easily form new hydrogen bonds with K71 and D74 in helix H3 (69-72), thereby inhibiting FBPase activity. Notably, both disulfiram and 212 might moderately reduce blood glucose output in vivo. Therefore, our current findings not only identify a new covalent allosteric site of FBPase but also establish a structural foundation and provide a promising way for the design of covalent allosteric drugs for glucose reduction.
摘要：
Fructose-1,6-bisphosphatase (FBPase), as a key rate-limiting enzyme in the gluconeogenesis (GNG) pathway, represents a practical therapeutic strategy for type 2 diabetes (T2D). Our previous work first identified cysteine residue 128 (C128) was an important allosteric site in the structure of FBPase, while pharmacologically targeting C128 attenuated the catalytic ability of FBPase. Herein, ten approved cysteine covalent drugs were selected for exploring FBPase inhibitory activities, and the alcohol deterrent disulfiram displayed superior inhibitory efficacy among those drugs. Based on the structure of lead compound disulfiram, 58 disulfide-derived compounds were designed and synthesized for investigating FBPase inhibitory activities. Optimal compound 3a exhibited significant FBPase inhibition and glucose-lowering efficacy in vitro and in vivo. Furthermore, 3a covalently modified the C128 site, and then regulated the N125-S124-S123 allosteric pathway of FBPase in mechanism. In summary, 3a has the potential to be a novel FBPase inhibitor for T2D therapy. (C) 2020 Elsevier Masson SAS. All rights reserved.
作者机构：
[Feng, Lingling; Xiao, Shan; Rao, Li; Wei, Lin; Hong, Zongqin; Wan, Jian; Ren, Yanliang; Wan, J] Cent China Normal Univ, Int Cooperat Base Pesticide & Green Synth Hubei, Key Lab Pesticide & Chem Biol CCNU, Minist Educ,Dept Chem, Wuhan 430079, Hubei, Peoples R China.
通讯机构：
[Ren, YL; Wan, J] C;Cent China Normal Univ, Int Cooperat Base Pesticide & Green Synth Hubei, Key Lab Pesticide & Chem Biol CCNU, Minist Educ,Dept Chem, Wuhan 430079, Hubei, Peoples R China.
关键词：
Aldolases;Algicides activities;De novo discovery;Fragment-based Virtual Screen;Thiourea
摘要：
By using a new Fragment-Based Virtual Screen strategy, two series of novel FBA-II inhibitors (thiourea derivatives) were de novo discovered based on the active site of fructose-1, 6-bisphosphate aldolase from Cyanobacterial (CyFBA). In comparison, most of the N-(2-benzoylhydrazine-1-carbonothioyl) benzamide derivatives (L14 approximately L22) exhibit higher CyFBA-II inhibitory activities compared to N-(phenylcarbamothioyl) benzamide derivatives (L1 approximately L13). Especially, compound L14 not only shows higher CyFBA-II activity (Ki=0.65muM), but also exhibits most potent in vivo activity against Synechocystis sp. PCC 6803 (EC50=0.09ppm), higher (7-fold) than that of our previous inhibitor (EC50=0.6ppm). The binding modes of compound L14 and CyFBA-II were further elucidated by jointly using DOX computational protocol, MM-PBSA and site-directed mutagenesis assays. The positive results suggest that strategy adopted in this study was promising to rapidly discovery the potent inhibitors with novel scaffolds. The satisfactory algicide activities suggest that the thiourea derivatives is very likely to be a promising lead for the development of novel specific algicides to solve Cyanobacterial harmful algal blooms (CHABs).
摘要：
Fructose-1, 6-bisphosphatase (FBPase) has been regarded as an attractive drug target to control blood glucose against Type 2 diabetes (T2D). In this study, by using the strategy of pharmacophore-based virtual screening, a novel scaffold inhibitor targeted the AMP allosteric site of human liver FBPase were screened, their inhibitory activities were further tested. The experimental results showed that compound H27 exhibited high inhibitory activities with the IC50 value of 5.3 mu M. Therefore, compound H27 was chosen as the probe molecule, it's possible binding conformation targeted into FBPase was identified by using DOX2.0 strategy. The importance of key residues (127, T31, K112 and R140) in allosteric site of FBPase for the binding inhibitors were validated by mutation experiments. The agreement between theory and experiment suggest that the interactional information of FBPase and inhibitors (H27) were reliable. On basis of these rational interactional information, the compound H29 was further designed to exhibit more potential FBPase inhibition (IC50 = 2.5 mu M). (C) 2018 Elsevier Inc. All rights reserved.
期刊：
European Journal of Medicinal Chemistry,2019年184:111749 ISSN：0223-5234
通讯作者：
Ren, Yanliang;Wan, Jian
作者机构：
[Feng, Lingling; Chen, Haifeng; Rao, Li; Wei, Lin; Ren, Yanliang; Wan, Jian; Han, Xinya; Xu, Yanhong; Song, Rongrong; Huang, Yunyuan] Cent China Normal Univ, Int Cooperat Base Pesticide & Green Synth Hubei, Key Lab Pesticide & Chem Biol CCNU, Minist Educ,Dept Chem, Wuhan 430079, Hubei, Peoples R China.;[Han, Xinya] Anhui Univ Technol, Sch Chem & Chem Engn, Maanshan 243002, Peoples R China.;[Chen, Haifeng] Beibu Gulf Univ, Ocean Coll, Qinzhou 535011, Guangxi, Peoples R China.;[Su, Chen; Peng, Chao] Natl Facil Prot Sci Shanghai, Zhangjiang Lab, Shanghai 201210, Peoples R China.
通讯机构：
[Ren, YL; Wan, J] C;Cent China Normal Univ, Int Cooperat Base Pesticide & Green Synth Hubei, Key Lab Pesticide & Chem Biol CCNU, Minist Educ,Dept Chem, Wuhan 430079, Hubei, Peoples R China.
关键词：
Covalent inhibitors;Fragment-based drug design;Fructose-1,6-bisphosphatase (FBPase);Novel allosteric site
摘要：
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 compounds 14b, 14c, 14i or 14n led 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.
期刊：
Journal of Chemical Information and Modeling,2017年57(6):1426-1438 ISSN：1549-9596
通讯作者：
Ren, Yanliang;Wan, Jian
作者机构：
[Feng, Lingling; Zhu, Shuaihua; Wan, Fen; Rao, Li; Wei, Lin; Hong, Zongqin; Ren, Yanliang; Wan, Jian; Zhu, Xiuyun; Han, Xinya; Peng, Hao] Cent China Normal Univ, Dept Chem, Minist Educ, Int Cooperat Base Pesticide & Green Synth Hubei,K, Wuhan 430079, Peoples R China.;[Guo, Li] Hubei Environm Monitoring Cent Stn, Wuhan 430072, Hubei, Peoples R China.
通讯机构：
[Ren, YL; Wan, J] C;Cent China Normal Univ, Dept Chem, Minist Educ, Int Cooperat Base Pesticide & Green Synth Hubei,K, Wuhan 430079, Peoples R China.
关键词：
FtsZ inhibitor;Trisubstituted benzimidazoles;QSAR;Docking studies;Molecular dynamics simulation;Binding model
摘要：
FtsZ, an essential cytokinesis protein, was a highly promising target for antibacterial agents. Following up the identification of trisubstituted benzimidazoles targeting Mycobacterium tuberculosis FtsZ (Mtb FtsZ) in previous studies and in order to explore the possible binding mode of these analogues, in silico methodologies such as 3D-QSAR, ProFunc analysis, molecular docking and molecular dynamic simulation were performed. They corroborated well with each other and gave credence to the proposed binding mechanism of trisubstituted benzimidazoles in the interdomain cleft region of Mtb FtsZ. The benzimidazole scaffold and cyclohexyl group of trisubstituted benzimidazoles were orthogonal to each other in low energy state and inclosed in a most hydrophobic environment formed by residues from the C-terminal beta sheets and H7 helix. The carbamate groups at the 5-position extended outward form the cleft cavity to the hydrophilic surface. The substituents at the 6-position fitted into the top portion of the cleft by directly interacting with the T7 loop. It was believed that the hydrophobic interactions and the polar contacts were major contributors to the stabilization of the ligand binding in the interdomain cleft. The binding mechanism provided useful clues to design new trisubstituted benzimidazoles inhibitors of Mtb FisZ with promising activity. The work presented here may even be expanded tremendously to screen novel Mtb FtsZ inhibitors with different scaffolds by using the molecular dynamic refined Mtb FtsZ structure and docking based 3D QSAR CoMFA.
