Ajdačić Vladimir, Senerovic Lidija, Vranić Marija, Pekmezovic Marina, Arsic-Arsnijevic Valentina, Veselinovic Aleksandar, Veselinovic Jovana, Šolaja Bogdan A, Nikodinovic-Runic Jasmina, Opsenica Igor M
Faculty of Chemistry, University of Belgrade, Studentski trg 16, PO Box 51, 11158 Belgrade, Serbia.
Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, 11000 Belgrade, Serbia.
Bioorg Med Chem. 2016 Mar 15;24(6):1277-91. doi: 10.1016/j.bmc.2016.01.058. Epub 2016 Feb 2.
A series of new thiophene-based guanylhydrazones (iminoguanidines) were synthesized in high yields using a straightforward two-step procedure. The antifungal activity of compounds was evaluated against a wide range of medicaly important fungal strains including yeasts, molds, and dermatophytes in comparison to clinically used drug voriconazole. Cytotoxic properties of compounds were also determined using human lung fibroblast cell line and hemolysis assay. All guanylhydrazones showed significant activity against broad spectrum of clinically important species of Candida spp., Aspergillus fumigatus, Fusarium oxysporum, Microsporum canis and Trichophyton mentagrophytes, which was in some cases comparable or better than activity of voriconazole. More importantly, compounds 10, 11, 13, 14, 18 and 21 exhibited excellent activity against voriconazole-resistant Candida albicans CA5 with very low minimal inhibitory concentration (MIC) values <2 μg mL(-1). Derivative 14, bearing bromine on the phenyl ring, was the most effective compound with MICs ranging from 0.25 to 6.25 μg mL(-1). However, bis-guanylhydrazone 18 showed better selectivity in terms of therapeutic index values. In vivo embryotoxicity on zebrafish (Danio rerio) showed improved toxicity profile of 11, 14 and 18 in comparison to that of voriconazole. Most guanylhydrazones also inhibited C. albicans yeast to hyphal transition, essential for its biofilm formation, while 11 and 18 were able to disperse preformed Candida biofilms. All guanylhydrazones showed the equal potential to interact with genomic DNA of C. albicans in vitro, thus indicating a possible mechanism of their action, as well as possible mechanism of observed cytotoxic effects. Tested compounds did not have significant hemolytic effect and caused low liposome leakage, which excluded the cell membrane as a primary target. On the basis of computational docking experiments using both human and cytochrome P450 from Candida it was concluded that the most active guanylhydrazones had minimal structural prerequisites to interact with the cytochrome P450 14α-demethylase (CYP51). Promising guanylhydrazone derivatives also showed satisfactory pharmacokinetic profile based on molecular calculations.
采用简单的两步法，以高产率合成了一系列新型噻吩基胍腙（亚氨基胍）。与临床使用的药物伏立康唑相比，评估了这些化合物对包括酵母、霉菌和皮肤癣菌在内的多种医学上重要的真菌菌株的抗真菌活性。还使用人肺成纤维细胞系和溶血试验测定了化合物的细胞毒性特性。所有胍腙对广泛的临床上重要的念珠菌属、烟曲霉、尖孢镰刀菌、犬小孢子菌和须癣毛癣菌均表现出显著活性，在某些情况下，其活性与伏立康唑相当或更好。更重要的是，化合物10、11、13、14、18和21对耐伏立康唑的白色念珠菌CA5表现出优异活性，其最低抑菌浓度（MIC）值极低，<2μg mL(-1)。在苯环上带有溴的衍生物14是最有效的化合物，MIC范围为0.25至6.25μg mL(-1)。然而，双胍腙18在治疗指数值方面表现出更好的选择性。对斑马鱼（Danio rerio）的体内胚胎毒性研究表明，与伏立康唑相比，11、14和18的毒性特征有所改善。大多数胍腙还抑制白色念珠菌从酵母向菌丝的转变，这对其生物膜形成至关重要，而11和18能够分散预先形成的念珠菌生物膜。所有胍腙在体外与白色念珠菌的基因组DNA相互作用的潜力相同，因此表明了它们可能的作用机制以及观察到的细胞毒性作用的可能机制。测试的化合物没有显著的溶血作用，且导致低脂质体泄漏，这排除了细胞膜作为主要靶点。基于使用人源和念珠菌细胞色素P450的计算对接实验得出结论，最具活性的胍腙与细胞色素P450 14α-脱甲基酶（CYP51）相互作用的结构先决条件最小。基于分子计算，有前景的胍腙衍生物也显示出令人满意的药代动力学特征。
