Direct modulation of cardiac myosin function has emerged as a therapeutic target for both heart disease and heart failure. However, the development of myosin-based therapeutics has been hampered by the lack of targeted in vitro screening assays. In this study we use Artificial Intelligence-based virtual high throughput screening (vHTS) to identify novel small molecule effectors of human β-cardiac myosin. We test the top scoring compounds from vHTS in biochemical counter-screens and identify a novel chemical scaffold called ‘F10’ as a cardiac-specific low-micromolar myosin inhibitor. Biochemical and biophysical characterization in both isolated proteins and muscle fibers show that F10 stabilizes both the biochemical (i.e. super-relaxed state) and structural (i.e. interacting heads motif) OFF state of cardiac myosin, and reduces force and left ventricular pressure development in isolated myofilaments and Langendorff-perfused hearts, respectively. F10 is a tunable scaffold for the further development of a novel class of myosin modulators.

直接调节心肌肌球蛋白功能已成为心脏病和心力衰竭的治疗靶点。然而，由于缺乏针对性的体外筛选试验，基于肌球蛋白的疗法的发展受到阻碍。在这项研究中，我们使用基于人工智能的虚拟高通量筛选（vHTS）来识别人类β-心肌肌球蛋白的新型小分子效应物。我们在生化反筛选中测试了 vHTS 中得分最高的化合物，并确定了一种称为“F10”的新型化学支架，作为心脏特异性低微摩尔肌球蛋白抑制剂。分离的蛋白质和肌纤维的生化和生物物理特征表明，F10 可以稳定心肌肌球蛋白的生化（即超松弛状态）和结构（即相互作用的头基序）OFF 状态，并减少分离的肌丝中的力和左心室压力发展和兰根多夫灌注的心脏，分别。 F10 是一种可调支架，用于进一步开发一类新型肌球蛋白调节剂。