Banfi Elena, Scialino Giuditta, Zampieri Daniele, Mamolo Maria Grazia, Vio Luciano, Ferrone Marco, Fermeglia Maurizio, Paneni Maria Silvia, Pricl Sabrina
Microbiology Laboratory, Department of Biomedical Sciences, University of Trieste I-34127 Trieste, Italy.
J Antimicrob Chemother. 2006 Jul;58(1):76-84. doi: 10.1093/jac/dkl182. Epub 2006 May 18.
To synthesize new antimycobacterial and antifungal drugs that act by binding to sterol 14alpha-demethylase (14DM) and to characterize the drug-target protein interactions using computer-based molecular simulations.
Different series of imidazole and triazole derivatives having an azomethine linkage to pyridine 2-carboxamidrazone were designed and synthesized. Molecular dynamic simulations of the sterol 14DM (a mixed-function oxidase involved in sterol synthesis in eukaryotic and prokaryotic organisms) complexed with new azole derivatives have been performed to both qualify and quantify the protein-ligand interactions. MICs of the compounds were evaluated by reference assay and by the recently developed Microdilution Resazurin Assay (MRA).
Halogenated derivatives showed good activity, with an MIC90 of 1 mg/L against 33 Candida spp. clinical strains; most compounds also had inhibitory activity against Mycobacterium tuberculosis reference and clinical strains, with MICs in the range 4-64 mg/L. Molecular modelling investigations showed that the active new compounds may interact at the active site of both the fungal and the mycobacterial cytochrome P450-dependent sterol-14alpha-demethylase and that the calculated binding free energy values are in agreement with the corresponding MIC values.
The combined experimental and computational approach can be helpful in targeted drug design, thus yielding valuable information for the synthesis and prediction of activity of a second generation of inhibitors.
合成通过与甾醇14α-去甲基酶（14DM）结合起作用的新型抗分枝杆菌和抗真菌药物，并使用基于计算机的分子模拟来表征药物与靶点蛋白的相互作用。
设计并合成了与吡啶2-羧脒腙具有甲亚胺键的不同系列的咪唑和三唑衍生物。对甾醇14DM（一种参与真核生物和原核生物甾醇合成的混合功能氧化酶）与新型唑类衍生物形成的复合物进行了分子动力学模拟，以鉴定和量化蛋白质-配体相互作用。通过参考测定法和最近开发的微量稀释刃天青测定法（MRA）评估化合物的最低抑菌浓度（MIC）。
卤代衍生物显示出良好的活性，对33株念珠菌临床菌株的MIC90为1mg/L；大多数化合物对结核分枝杆菌参考菌株和临床菌株也具有抑制活性，MIC范围为4-64mg/L。分子建模研究表明，活性新化合物可能在真菌和分枝杆菌细胞色素P450依赖性甾醇-14α-去甲基酶的活性位点相互作用，并且计算出的结合自由能值与相应的MIC值一致。
实验和计算相结合的方法有助于靶向药物设计，从而为第二代抑制剂的合成和活性预测提供有价值的信息。
