Them1 Inhibitors for the Management of Non-Alcoholic Fatty Liver Disease

项目摘要/摘要 对于非酒精性脂肪肝的治疗，迫切需要有效的药物治疗。 (NAFLD)。拟议的研究是对新的治疗线索进行早期临床前验证， 靶向硫酯酶超家族成员1(Them1；同义词酰基辅酶11)，脂肪的关键酶 酸代谢在NAFLD中变得不适应。长期目标是开发Them1 AS的抑制剂 治疗人类非酒精性脂肪肝的一种方法。这项研究的目的是优化 小分子抑制剂的类药物性质，以生成先导化合物，并展示其 在基于细胞的分析和使用小鼠的实验性NAFLD中的安全性、有效性和特异性。目标主题1 使用优化的小分子抑制剂有望通过以下方式缓解NAFLD：1)增加能量消耗 产热的棕色和米色脂肪组织；2)减少肝脏脂肪变性和葡萄糖的产生 通过肝脏；以及3)减少白色脂肪组织的炎症。其基本原理是一种先导化合物 解决了Them1的这三个独立的致病贡献，应该证明在NAFLD中是有效的。我们 已经完成了高通量的小分子筛查，已经确定了有希望的靶向 Them1.脂肪酰辅酶A硫代酯酶活性动机是关于以下贡献的大量机械数据 对于NAFLD的发病机制，抑制剂向先导化合物的开发将在 三个具体目标：1)优化Them1小分子抑制剂的效力和特异性；2) 建立早期先导化合物在细胞培养中的类药物特性和疗效；以及3)评估 实验性非酒精性脂肪肝小鼠体内的治疗性先导化合物。在目标1中，小分子抑制剂将是 通过药物化学策略进行优化，以提高效力、特异性和类药物特性。这些 这些努力将以基于体外酶分析的结构-活性关系以及 Them1-抑制剂相互作用的生物物理和结构决定因素。AIM 2将评估早期销售线索 具有类药物特性和细胞毒性的化合物。然后将对选定的早期导联进行疗效测试 在原代培养的小鼠和人类细胞中增加棕色脂肪酸氧化的能力 脂肪细胞和肝细胞，以减少肝细胞中肝葡萄糖的产生和减少产生 白色脂肪细胞的炎性介质。特异性将使用从Them1-/-培养的细胞进行评估 老鼠。在目标3中，将在小鼠身上测试先导化合物的药代动力学和耐受性。在建立 给药路线和时间表，预防和逆转NAFLD的有效性将在小鼠模型中进行评估。 偏离目标的效果将使用Them1-/-小鼠进行评估。这些研究的预期结果是 优化的Lead Them1抑制剂的开发和临床前验证，该抑制剂利用新的机制 能量平衡、脂肪酸和葡萄糖代谢，以及医疗管理中的炎症 非酒精性脂肪肝。

PROJECT SUMMARY/ABSTRACT There is a critical need for effective pharmacotherapy for the management of non-alcoholic fatty liver disease (NAFLD). The proposed research is to conduct early-stage preclinical validation of new therapeutic leads that target thioesterase superfamily member 1 (Them1; synonym Acyl-CoA thioesterase 11), a key enzyme of fatty acid metabolism that becomes maladaptive in NAFLD. The long-term goal is to develop inhibitors of Them1 as a therapeutic modality in the management of human NAFLD. The objective of this research is to optimize the drug-like properties of small molecule inhibitors in order to create lead compounds, and to demonstrate their safety, efficacy and specificity in cell-based assays and in experimental NAFLD using mice. Targeting Them1 with optimized small molecule inhibitors is expected to mitigate NAFLD by: 1) Increasing energy expenditure in thermogenic brown and beige adipose tissue; 2) Decreasing hepatic steatosis, as well as glucose production by the liver; and 3) Reducing inflammation in white adipose tissue. The rationale is that a lead compound that addresses these three independent pathogenic contributions of Them1 should prove effective in NAFLD. We have completed a high-throughput small molecule screen that has identified promising inhibitors targeting the fatty acyl-CoA thioesterase activity of Them1. Motivated by extensive mechanistic data on the contributions of Them1 to NAFLD pathogenesis, the development of inhibitors into lead compounds will be accomplished in three specific aims: 1) To optimize the potency and specificity of Them1 small molecule inhibitors; 2) To establish drug-like properties and efficacy in cell culture of early lead compounds; and 3) To evaluate therapeutic lead compounds in experimental NAFLD using mice. In Aim 1, small molecule inhibitors will be optimized by medicinal chemistry strategies to improve potency, specificity and drug-like properties. These efforts will be guided by structure-activity relationships based on in vitro enzymatic assays, as well as biophysical and structural determinants of Them1-inhibitor interactions. Aim 2 will evaluate early lead compounds for drug-like properties, as well as cytotoxicity. Selected early leads will then be tested for efficacy in primary cultured mouse and human cells for their capacities to increase fatty acid oxidation in brown adipocytes and hepatocytes, to reduce hepatic glucose production in hepatocytes and to decrease production of inflammatory mediators by white adipocytes. Specificity will be assessed using cells cultured from Them1-/- mice. In Aim 3, lead compounds will be tested in mice for pharmacokinetics and tolerability. Upon establishing dosing routes and schedules, efficacy to prevent and to reverse NAFLD will be assessed in mouse models. Off-target effects will be evaluated using Them1-/- mice. The expected outcome of these studies is the development and pre-clinical validation of optimized lead Them1 inhibitors that leverage novel mechanisms of energy homeostasis, fatty acid and glucose metabolism, as well as inflammation in the medical management of NAFLD.