title: Screening of Pharmacologically Active Small Molecule Compounds Identifies Antifungal Agents Against Candida Biofilms.
abstract: 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.
authors.affiliation: Department of Applied Prosthodontics, Graduate School of Biomedical Sciences, Nagasaki University Nagasaki, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Watamoto
authors.affiliation: Division of Molecular and Regenerative Prosthodontics, Tohoku University Graduate School of Dentistry Sendai, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Egusa
authors.affiliation: Department of Applied Prosthodontics, Graduate School of Biomedical Sciences, Nagasaki University Nagasaki, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Sawase
authors.affiliation: Department of Fixed Prosthodontics, Osaka University Graduate School of Dentistry Suita, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Yatani
keywords: Candida albicans; Small molecules; antifungal drug; biofilm; drug discovery