Watamoto Takao, Egusa Hiroshi, Sawase Takashi, Yatani Hirofumi
Department of Applied Prosthodontics, Graduate School of Biomedical Sciences, Nagasaki University Nagasaki, Japan.
Division of Molecular and Regenerative Prosthodontics, Tohoku University Graduate School of Dentistry Sendai, Japan.
Front Microbiol. 2015 Dec 22;6:1453. doi: 10.3389/fmicb.2015.01453. eCollection 2015.
Candida species have emerged as important and common opportunistic human pathogens, particularly in immunocompromised individuals. The current antifungal therapies either have toxic side effects or are insufficiently effect. The aim of this study is develop new small-molecule antifungal compounds by library screening methods using Candida albicans, and to evaluate their antifungal effects on Candida biofilms and cytotoxic effects on human cells. Wild-type C. albicans strain SC5314 was used in library screening. To identify antifungal compounds, we screened a small-molecule library of 1,280 pharmacologically active compounds (LOPAC(1280TM)) using an antifungal susceptibility test (AST). To investigate the antifungal effects of the hit compounds, ASTs were conducted using Candida strains in various growth modes, including biofilms. We tested the cytotoxicity of the hit compounds using human gingival fibroblast (hGF) cells to evaluate their clinical safety. Only 35 compounds were identified by screening, which inhibited the metabolic activity of C. albicans by >50%. Of these, 26 compounds had fungistatic effects and nine compounds had fungicidal effects on C. albicans. Five compounds, BAY11-7082, BAY11-7085, sanguinarine chloride hydrate, ellipticine and CV-3988, had strong fungicidal effects and could inhibit the metabolic activity of Candida biofilms. However, BAY11-7082, BAY11-7085, sanguinarine chloride hydrate and ellipticine were cytotoxic to hGF cells at low concentrations. CV-3988 showed no cytotoxicity at a fungicidal concentration. Four of the compounds identified, BAY11-7082, BAY11-7085, sanguinarine chloride hydrate and ellipticine, had toxic effects on Candida strains and hGF cells. In contrast, CV-3988 had fungicidal effects on Candida strains, but low cytotoxic effects on hGF cells. Therefore, this screening reveals agent, CV-3988 that was previously unknown to be antifungal agent, which could be a novel therapies for superficial mucosal candidiasis.
念珠菌属已成为重要且常见的人类机会致病菌，尤其是在免疫功能低下的个体中。目前的抗真菌疗法要么具有毒副作用，要么效果不佳。本研究的目的是通过使用白色念珠菌的文库筛选方法开发新的小分子抗真菌化合物，并评估它们对念珠菌生物膜的抗真菌作用以及对人类细胞的细胞毒性作用。野生型白色念珠菌菌株SC5314用于文库筛选。为了鉴定抗真菌化合物，我们使用抗真菌药敏试验（AST）筛选了一个包含1280种药理活性化合物的小分子文库（LOPAC(1280TM)）。为了研究筛选出的化合物的抗真菌作用，我们使用包括生物膜在内的各种生长模式的念珠菌菌株进行了AST试验。我们使用人牙龈成纤维细胞（hGF）测试了筛选出的化合物的细胞毒性，以评估它们的临床安全性。通过筛选仅鉴定出35种化合物，这些化合物抑制白色念珠菌的代谢活性超过50%。其中，26种化合物对白色念珠菌有抑菌作用，9种化合物有杀菌作用。五种化合物，BAY11-7082、BAY11-7085、水合氯化血根碱、玫瑰树碱和CV-3988，具有很强的杀菌作用，并且可以抑制念珠菌生物膜的代谢活性。然而，BAY11-7082、BAY11-7085、水合氯化血根碱和玫瑰树碱在低浓度下对hGF细胞具有细胞毒性。CV-3988在杀菌浓度下未显示细胞毒性。鉴定出的四种化合物，BAY11-7082、BAY11-7085、水合氯化血根碱和玫瑰树碱，对念珠菌菌株和hGF细胞有毒性作用。相比之下，CV-3988对念珠菌菌株有杀菌作用，但对hGF细胞的细胞毒性较低。因此，这次筛选揭示了一种此前未知的抗真菌药物CV-3988，它可能是治疗浅表黏膜念珠菌病的一种新疗法。
