Tissue repair and regenerative medicine address the important medical needs to replace damaged tissue with functional tissue. Most regenerative medicine strategies have focused on delivering biomaterials and cells, yet there is the untapped potential for drug-induced regeneration with good specificity and safety profiles. The Hippo pathway is a key regulator of organ size and regeneration by inhibiting cell proliferation and promoting apoptosis. Kinases MST1 and MST2 (MST1/2), the mammalian Hippo orthologs, are central components of this pathway and are, therefore, strong target candidates for pharmacologically induced tissue regeneration. We report the discovery of a reversible and selective MST1/2 inhibitor, 4-((5,10-dimethyl-6-oxo-6,10-dihydro-5H-pyrimido[5,4-b]thieno[3,2-e][1,4]diazepin-2-yl)amino)benzenesulfonamide (XMU-MP-1), using an enzyme-linked immunosorbent assay–based high-throughput biochemical assay. The cocrystal structure and the structure-activity relationship confirmed that XMU-MP-1 is on-target to MST1/2. XMU-MP-1 blocked MST1/2 kinase activities, thereby activating the downstream effector Yes-associated protein and promoting cell growth. XMU-MP-1 displayed excellent in vivo pharmacokinetics and was able to augment mouse intestinal repair, as well as liver repair and regeneration, in both acute and chronic liver injury mouse models at a dose of 1 to 3 mg/kg via intraperitoneal injection. XMU-MP-1 treatment exhibited substantially greater repopulation rate of human hepatocytes in the Fah-deficient mouse model than in the vehicle-treated control, indicating that XMU-MP-1 treatment might facilitate human liver regeneration. Thus, the pharmacological modulation of MST1/2 kinase activities provides a novel approach to potentiate tissue repair and regeneration, with XMU-MP-1 as the first lead for the development of targeted regenerative therapeutics.

组织修复和再生医学满足了用功能组织替代受损组织的重要医学需求。大多数再生医学策略都集中在传递生物材料和细胞上，但是具有良好的特异性和安全性的药物诱导的再生仍有未开发的潜力。通过抑制细胞增殖和促进细胞凋亡，Hippo途径是器官大小和再生的关键调节剂。哺乳动物河马直系同源蛋白激酶MST1和MST2（MST1 / 2）是该途径的重要组成部分，因此，是药理学诱导的组织再生的强靶标候选物。我们报告发现了可逆和选择性的MST1 / 2抑制剂4-（（（5,10-二甲基-6-氧代-6,10-二氢-5H-嘧啶[5,4-b] thieno [3,2- e] [1,4]二氮杂-2-（基）氨基）苯磺酰胺（XMU-MP-1），使用基于酶联免疫吸附测定的高通量生化测定。共晶体结构和结构-活性关系证实，XMU-MP-1是MST1 / 2的靶标。XMU-MP-1阻断MST1 / 2激酶活性，从而激活下游效应子Yes相关蛋白并促进细胞生长。XMU-MP-1在腹腔注射剂量为1至3 mg / kg的急性和慢性肝损伤小鼠模型中均显示出优异的体内药代动力学，并能够增强小鼠肠道的修复以及肝脏的修复和再生。XMU-MP-1处理在肝细胞中显示出更高的人类肝细胞再增殖率。共晶体结构和结构-活性关系证实，XMU-MP-1是MST1 / 2的靶标。XMU-MP-1阻断MST1 / 2激酶活性，从而激活下游效应子Yes相关蛋白并促进细胞生长。XMU-MP-1在腹腔注射剂量为1至3 mg / kg的急性和慢性肝损伤小鼠模型中均显示出优异的体内药代动力学，并能够增强小鼠肠道的修复以及肝脏的修复和再生。XMU-MP-1处理在肝细胞中显示出更高的人类肝细胞再增殖率。共晶体结构和结构-活性关系证实，XMU-MP-1是MST1 / 2的靶标。XMU-MP-1阻断MST1 / 2激酶活性，从而激活下游效应子Yes相关蛋白并促进细胞生长。XMU-MP-1在腹腔注射剂量为1至3 mg / kg的急性和慢性肝损伤小鼠模型中均显示出优异的体内药代动力学，并能够增强小鼠肠道的修复以及肝脏的修复和再生。XMU-MP-1处理在肝细胞中显示出更高的人类肝细胞再增殖率。以及在急性和慢性肝损伤小鼠模型中，通过腹膜内注射以1至3 mg / kg的剂量进行肝修复和再生。XMU-MP-1处理在肝细胞中显示出更高的人类肝细胞再增殖率。以及在急性和慢性肝损伤小鼠模型中，通过腹膜内注射以1至3 mg / kg的剂量进行肝修复和再生。XMU-MP-1处理在肝细胞中显示出更高的人类肝细胞再增殖率。Fah缺陷型小鼠模型比赋形剂处理的对照组要好，这表明XMU-MP-1处理可能促进人肝再生。因此，MST1 / 2激酶活性的药理学调节提供了一种增强组织修复和再生的新方法，其中XMU-MP-1作为开发靶向再生疗法的第一个先导。