Identification and validation of the mode of action of the chalcone anti-mycobacterial compounds.
Identification and validation of the mode of action of the chalcone anti-mycobacterial compounds.
复制标题
查耳酮抗分枝杆菌化合物作用方式的鉴定和验证。
DOI：
10.1016/j.tcsw.2020.100041
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Anagani B
中科院分区：
文献类型：
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作者：
Anagani B
目的寻找新的抗结核药物已成为现代药物化学面临的重大挑战之一。通过开发新的、更短的、廉价的、安全有效的抗TB方案，可以改善TB化疗的结果。方法合成查尔酮（1a-1 o），并使用生长抑制试验评估其抗分枝杆菌活性。化合物1a被选为“命中”化合物。化合物1a的作用模式通过使用薄层色谱法的分枝菌酸甲酯（MAME）和脂肪酸甲酯（FAME）分析来鉴定。通过过量表达FAS-Ⅱ的组分进行剂量依赖性实验。bovisBCG来确认目标。配体结合使用固有色氨酸测定和分子对接被用来进一步验证target.ResultsMAMEs和FAMEs分析表明，剂量依赖性减少MAMEs与FAMEs的总体丰度表明，compound 1 atargets分枝菌酸生物合成。直接绑定of 1ato InhA观察使用固有的色氨酸荧光结合试验，和2倍的IC 50偏移与InhA过表达菌株确认InhA作为cellular target.ConclusionThe查耳酮1a表现出有效的抗分枝杆菌活性，显示出良好的安全性，是一个直接抑制剂的InhA，分枝菌酸合成的关键组成部分，验证这一系列进一步的抗TB药物的开发。
ObjectivesThe search for new TB drugs has become one of the great challenges for modern medicinal chemistry. An improvement in the outcomes of TB chemotherapy can be achieved by the development of new, shorter, cheap, safe and effective anti-TB regimens.MethodsChalcones (1a-1o) were synthesized and evaluated for their antimycobacterial activity againstMycobacterium bovisBCG using growth inhibition assays. Compound1awas selected as a ‘hit’ compound. The mode of action of compound1a, was identified by mycolic acid methyl esters (MAMEs) and fatty acid methyl esters (FAMEs) analysis using thin layer chromatography. Dose dependent experiments were conducted by over-expressing components of FAS-II inM. bovisBCG to confirm the target. Ligand binding using intrinsic tryptophan assay and molecular docking were used to further validate the target.ResultsMAMEs and FAMEs analysis showed dose-dependent reduction of MAMEs with the overall abundance of FAMEs suggesting that compound1atargets mycolic acid biosynthesis. Direct binding of1ato InhA was observed using an intrinsic tryptophan fluorescence binding assay, and a 2-fold IC50shift was observed with an InhA overexpressing strain confirming InhA as the cellular target.ConclusionThe chalcone1aexhibits potent antimycobacterial activity, displays a good safety profile and is a direct inhibitor of InhA, a key component in mycolic acid synthesis, validating this series for further anti-TB drug development.