Axin Stabilization by Novel Small Molecules to Treat Non-alcoholic Steatohepatitis

项目总结/摘要 非酒精性脂肪性肝炎（NASH）影响了12%的美国人，因为越来越多的人 代谢紊乱，包括非酒精性脂肪肝、肥胖、代谢综合征和糖尿病。 目前，尚无批准用于NASH的药物选择。Wnt/β-catenin通路与腺苷 单磷酸活化激酶（AMPK）是调节肝脏代谢的重要途径。遗传和 药理学证据支持通过抑制Wnt/β-连环蛋白对肝代谢的益处; 同时，AMPK激活已成为对抗NASH的机制。我们合成并测试了一种 新型化合物，其中三唑YW 1128显示出Wnt/β-连环蛋白抑制和 AMPK通过Axin稳定机制激活。YW 1128显示出令人兴奋的抗肝细胞癌的功效， 在体外和在小鼠中的脂肪变性。基于YW 1128的化学结构，我们最近设计并 合成了新的铅YA 6060。在保持双重活动的同时，YA 6060表现出了非常有前途的 药物样特征，特别是抗肝脂肪变性和纤维化的令人兴奋的功效。我们还发现 端锚聚合酶结合蛋白1（TNKS 1bp 1或TAB 182）作为我们的先导化合物的结合靶标。的 拟进行的研究旨在验证同时抑制Wnt/β-连环蛋白通路 通过用小分子靶向TAB 182稳定细胞Axin水平来激活AMPK，是一种新的 NASH治疗的治疗策略。为了验证这一假设，我们将首先综合和评估新的 抑制Wnt/β-连环蛋白信号传导并激活AMPK的化合物，使用涉及新的 类似物设计、合成化学、药物性质的体外评估和体内药代动力学， 毒性评价第二，通过进行包括配体结合测定的体外和体内实验， 基因敲除、转基因表达、基因突变、化学抑制或激活，以及各种 通过功能测定，我们将确定铅化合物在肝细胞和小鼠中的作用机制。 最后，我们将评估YA 6060和一种新的基于YA 6060的先导化合物的功效，并验证它们的有效性。 在具有全范围NASH适应症的小鼠模型中的药理学机制。成功完成 该项目有望建立具有Wnt/β-连环蛋白抑制和 通过靶向TAB 182和Axin稳定化激活AMPK是NASH发展的有效资源 治疗学这些知识和小分子预计将大大有利于未来的研究， 了解NASH发病机制。最终，我们希望通过一流的医疗服务来改变NASH患者的护理。 药

PROJECT SUMMARY/ABSTRACT Non-alcoholic steatohepatitis (NASH) affects 12% of Americans due to the growing number of individuals with metabolic disorders, including nonalcoholic fatty liver disease, obesity, metabolic syndromes, and diabetes. At present, there are no pharmacological options approved for NASH. Wnt/beta-catenin pathway and adenosine monophosphate-activated kinase (AMPK) are crucial pathways in regulation of hepatic metabolism. Genetic and pharmacological evidence supports a benefit by inhibition of Wnt/beta-catenin on hepatic metabolism; meanwhile, AMPK activation has emerged as a mechanism against NASH. We have synthesized and tested a novel class of compounds, of which triazole YW1128 showed dual activities of Wnt/beta-catenin inhibition and AMPK activation via the mechanism of Axin stabilization. YW1128 showed an exciting efficacy against hepatic steatosis both in vitro and in mice. Based on the chemical structure of YW1128, we recently designed and synthesized the new lead YA6060. While maintaining the dual activities, YA6060 has exhibited a highly promising drug-like profile and particularly an exciting efficacy against hepatic steatosis and fibrosis. We also discovered the tankyrase-binding protein 1 (TNKS1bp1 or TAB182) as a binding target of our lead compounds. The proposed studies are intended to test the hypothesis that simultaneously inhibiting Wnt/beta-catenin pathway and activating AMPK, by stabilizing cellular Axin level via targeting TAB182 with small molecules, is a novel therapeutic strategy for NASH treatment. To test this hypothesis, we will first synthesize and evaluate new compounds that inhibit Wnt/β-catenin signaling and activate AMPK, using an iterative process involving new analog design, synthetic chemistry, in vitro assessment of drug properties, and in vivo pharmacokinetic and toxicity evaluation. Second, by conducting in vitro and in vivo experiments including ligand binding assays, genetic knockdown, transgene expression, genetic mutation, chemical inhibition or activation, and various functional assays, we will determine the mechanism of action for lead compounds in hepatocytes and mice. Lastly, we will evaluate the efficacy of YA6060 and a new YA6060-based lead compound and validate their pharmacological mechanism in mouse models with a full range of NASH indications. Successful completion of this project is expected to establish that small molecules with dual activities of Wnt/beta-catenin inhibition and AMPK activation via targeting TAB182 and Axin stabilization is a valid resource for the development of NASH therapeutics. The knowledge and small molecules are expected to greatly benefit future research in understanding of NASH pathogenesis. Eventually, we expect to transform NASH patient care with a first-in-class drug.