Ildefeld Niklas, Steinhilber Dieter, Proschak Ewgenij, Heering Jan
Institute of Pharmaceutical Chemistry, Goethe-University of Frankfurt, Biocenter, Max-von-Laue-Str. 9, 60438 Frankfurt/Main, Germany.
Fraunhofer Institute for Translational Medicine and Pharmacology ITMP, Theodor-Stern-Kai 7, 60596 Frankfurt/Main, Germany.
iScience. 2024 Jun 20;27(7):110333. doi: 10.1016/j.isci.2024.110333. eCollection 2024 Jul 19.
The COVID-19 pandemic has highlighted the lack of effective, ready-to-use antivirals for the treatment of viruses with pandemic potential. The development of a diverse drug portfolio is therefore crucial for pandemic preparedness. Viral macrodomains are attractive therapeutic targets as they are suggested to play an important role in evading the innate host immune response, making them critical for viral pathogenesis. Macrodomains function as erasers of mono-ADP-ribosylation (deMARylation), a post-translational modification that is involved in interferon signaling. Herein, we report the development of a modular HTRF-based assay, that can be used to screen for inhibitors of various viral and human macrodomains. We characterized the five most promising small molecule SARS-CoV-2 Mac1 inhibitors recently reported in the literature for potency and selectivity and conducted a pilot screen demonstrating HTS suitability. The ability to directly detect enzymatic activity makes the DeMAR assay a valuable addition to the existing tools for macrodomain drug discovery.
新冠疫情凸显了缺乏用于治疗具有大流行潜力病毒的有效即用型抗病毒药物的问题。因此，开发多样化的药物组合对于大流行防范至关重要。病毒大结构域是有吸引力的治疗靶点，因为它们被认为在逃避宿主固有免疫反应中起重要作用，这使其对病毒发病机制至关重要。大结构域作为单磷酸腺苷核糖基化的擦除酶（去单磷酸腺苷核糖基化）发挥作用，单磷酸腺苷核糖基化是一种参与干扰素信号传导的翻译后修饰。在此，我们报告了一种基于模块化HTRF的检测方法的开发，该方法可用于筛选各种病毒和人类大结构域的抑制剂。我们对文献中最近报道的五种最有前景的小分子SARS-CoV-2 Mac1抑制剂的效力和选择性进行了表征，并进行了初步筛选以证明其适用于高通量筛选。直接检测酶活性的能力使去单磷酸腺苷核糖基化检测成为大结构域药物发现现有工具的宝贵补充。
