Wei Kaiyao, Arlotto Marie, Overhulse Justin M, Dinh Tuan-Anh, Zhou Yingsheng, Dupper Nathan J, Yang Jiayi, Kashemirov Boris A, Dawi Hasan, Garnaud Cécile, Bourgine Gaëlle, Mietton Flore, Champleboux Morgane, Larabi Amédé, Hayat Yordan, Indorato Rose-Laure, Noirclerc-Savoye Marjolaine, Skoufias Dimitrios, Cornet Muriel, Rabut Gwenaël, McKenna Charles E, Petosa Carlo, Govin Jérôme
Univ. Grenoble Alpes, CEA, CNRS, Institut de Biologie Structurale (IBS), Grenoble, 38000, France.
Univ. Grenoble Alpes, Inserm, CNRS, Institute for Advanced Biosciences (IAB), Grenoble, 38000, France.
Adv Sci (Weinh). 2025 Mar;12(10):e2404260. doi: 10.1002/advs.202404260. Epub 2025 Jan 16.
The fungal Bromodomain and Extra-Terminal (BET) protein Bdf1 is a potential antifungal target against invasive fungal infections. However, the need to selectively inhibit both Bdf1 bromodomains (BDs) over human orthologs and the lack of molecular tools to assess on-target antifungal efficacy hamper efforts to develop Bdf1 BD inhibitors as antifungal therapeutics. This study reports a phenyltriazine compound that inhibits both Bdf1 BDs from the human fungal pathogen Candida glabrata with selectivity over the orthologous BDs from the human BET protein Brd4. On-target antifungal activity is established by devising two yeast-based inhibition assays: a growth assay using humanized Candida strains in which the Bdf1 BDs are replaced by their Brd4 counterparts, and a NanoBiT assay that evaluates the BD-mediated association of Bdf1 with chromatin. These assays additionally enable the discovery that BET inhibitor I-BET726 targets both Bdf1 BDs, inhibits the growth of a broad spectrum of Candida species, including antifungal-resistant clinical isolates, and displays efficacy in an invertebrate animal model of infection. These collective findings highlight the promising potential of Bdf1 BD inhibitors as an innovative class of antifungal therapeutics and the pivotal role of yeast-based assay development toward achieving this end.
真菌的溴结构域和额外末端（BET）蛋白Bdf1是针对侵袭性真菌感染的潜在抗真菌靶点。然而，需要在人同源物上选择性抑制Bdf1的两个溴结构域（BDs），且缺乏评估靶向抗真菌疗效的分子工具，这阻碍了将Bdf1 BD抑制剂开发为抗真菌治疗药物的努力。本研究报道了一种苯基三嗪化合物，它能抑制来自人类真菌病原体光滑念珠菌的Bdf1两个BDs，且对来自人类BET蛋白Brd4的同源BDs具有选择性。通过设计两种基于酵母的抑制试验来确定靶向抗真菌活性：一种是使用人源化念珠菌菌株的生长试验，其中Bdf1的BDs被其Brd4对应物取代；另一种是评估BD介导的Bdf1与染色质结合的纳米生物发光互补分析（NanoBiT分析）。这些试验还发现BET抑制剂I-BET726靶向Bdf1的两个BDs，抑制包括耐抗真菌临床分离株在内的多种念珠菌的生长，并在无脊椎动物感染模型中显示出疗效。这些共同发现突出了Bdf1 BD抑制剂作为一类创新抗真菌治疗药物的广阔前景，以及基于酵母的试验开发对实现这一目标的关键作用。
