Benzoates as toxic intermediate are naturally produced by fungal intracellular metabolism, and CYP53 can specific transform the substrates. In the study, we constructed the CYP53 homology model and analyzed the corresponding active region. At the same time, the molecular docking and the structure-based pharmacophore model (SBP) were performed to explore the bind mode of representative CYP53 inhibitors. On the basis, a series of phenylpyridines derivatives were designed as novel CYP53 inhibitors, and their molecular structures were synthesized and evaluated. Compared with the positive control groups, their antifungal activity showed the obvious upward trend. In particular, target compounds (13a, 15b) possessed the excellent biological activity against pathogenic fungi and drug-resistant fungi in vivo and in vitro. The preliminary action mechanism has confirmed that target compounds could inhibit CYP53 activity, and block the metabolism of toxic intermediates (Benzoates). This further induced the accumulation of reactive oxygen species (ROS) through the pattern of mitochondrial depolarization, which eventually caused fungal lysis and death. In summary, the study provided the reasonable computational models, and effectively guided the generation of novel CYP53 antifungal inhibitors.

苯甲酸盐作为有毒中间体是由真菌细胞内代谢自然产生的，CYP53可以特异性转化底物。在研究中，我们构建了CYP53同源模型并分析了相应的活性区域。同时，通过分子对接和基于结构的药效团模型（SBP）来探索代表性CYP53抑制剂的结合模式。在此基础上，设计了一系列苯基吡啶类衍生物作为新型CYP53抑制剂，并对其分子结构进行了合成和评价。与阳性对照组相比，其抗真菌活性呈明显上升趋势。尤其是目标化合物( 13a, 15b ) 具有优异的抗病原真菌和耐药真菌的生物活性。体内和体外。初步作用机制证实，靶标化合物可抑制CYP53活性，阻断有毒中间体（苯甲酸盐）的代谢。这进一步通过线粒体去极化模式诱导活性氧（ROS）的积累，最终导致真菌溶解和死亡。综上所述，该研究提供了合理的计算模型，有效指导了新型CYP53抗真菌抑制剂的产生。