The small molecule blebbistatin is now a front line tool in the study of myosin function. Chemical modification of the tricyclic core of blebbistatin could deliver the next generation of myosin inhibitors and to help address this we report here on the impact of structural changes in the methyl-substituted aromatic ring of blebbistatin on its biological activity. Chemical methods for the preparation of isomeric methyl-containing analogues are reported and a series of co-crystal structures are used to rationalise the observed variations in their biological activity. These studies further support the view that the previously identified binding mode of blebbistatin to Dictyostelium discoideum myosin II is of relevance to its mode of action. A discussion of the role that these observations have on planning the synthesis of focused libraries of blebbistatin analogues is also provided including an assessment of possibilities by computational methods. These studies are ultimately directed at the development of novel myosin inhibitors with improved affinity and different selectivity profiles from blebbistatin itself.

中文翻译：

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小分子工具 (S)-(-)-blebbistatin：与肌球蛋白抑制剂设计相关的新见解。

小分子 blebbistatin 现在是研究肌球蛋白功能的前沿工具。blebbistatin 三环核心的化学修饰可以提供下一代肌球蛋白抑制剂，为了帮助解决这个问题，我们在此报告了 blebbistatin 的甲基取代芳环结构变化对其生物活性的影响。报道了制备含甲基异构体类似物的化学方法，并使用一系列共晶结构来合理化观察到的生物活性变化。这些研究进一步支持了先前确定的 blebbistatin 与盘基网柄菌肌球蛋白 II 的结合模式与其作用模式相关的观点。还讨论了这些观察结果对计划合成重点布雷他汀类似物文库的作用，包括通过计算方法评估可能性。这些研究最终旨在开发与 blebbistatin 本身具有更高亲和力和不同选择性的新型肌球蛋白抑制剂。