Targeting the orphan nuclear receptor LRH-1 with small molecules

用小分子靶向孤儿核受体 LRH-1

• 批准号: 10681892
• 负责人: John Winter Calvert
• 金额: $ 15.4万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10681892
• 关键词: Address; Adipose tissue; Affinity; Agonist; American; Antidiabetic Drugs; Atherosclerosis; Award; Behavior; Binding; Binding Sites; Biological; Biological Availability; Biology; CRISPR/Cas technology; Cardiovascular Diseases; Characteristics; Charge; Chemicals; Chemistry; Clinical; Development; Diabetes Mellitus; Disease; Dose; Drug Kinetics; Epidemic; Exposure to; Fatty acid glycerol esters; Gene Expression; Generations; Glucose; Health; Homeostasis; Human; Insulin; Insulin Resistance; Knowledge; Lead; Ligand Binding; Ligands; Lipids; Liver; Measures; Metabolic; Metabolic Diseases; Metabolic Pathway; Modeling; Modification; Mus; Myocardial Infarction; NR5A2 gene; Non-Insulin-Dependent Diabetes Mellitus; Nuclear Hormone Receptors; Nuclear Orphan Receptor; Obese Mice; Obesity; Oral cavity; Overweight; Pharmacodynamics; Pharmacology; Phospholipids; Plant Roots; Plasma; Research; Risk; Rodent; Rodent Model; Stroke; Structure; Structure-Activity Relationship; Surface; Testing; Therapeutic; Therapeutic Agents; Tissues; United States; Work; base; biophysical properties; comorbidity; design; diet-induced obesity; dietary; efficacy evaluation; fatty liver disease; glucose metabolism; glucose tolerance; humanized mouse; improved; in vivo; innovation; insulin sensitivity; lipid metabolism; lipophilicity; liver function; mRNA Expression; mortality risk; mouse model; new therapeutic target; non-alcoholic fatty liver disease; novel; nutrition; pre-clinical; preclinical study; reverse cholesterol transport; small molecule; success; tool; transcriptome sequencing

Obesity is a growing epidemic in the United States, leading to increases in cases of nonalcoholic fatty liver disease (NAFLD), cardiovascular disease, and type II diabetes. A common characteristic of these diseases is aberrant lipid and glucose metabolism. This proposal centers on the nuclear hormone receptor, Liver Receptor Homolog 1 (LRH-1), which acts as an important regulator of lipid metabolism, reverse cholesterol transport, glucose sensing, and homeostasis. As such, LRH-1 represents a novel therapeutic target for metabolic diseases. LRH-1 binds to phospholipids (PLs) and is activated by the unusual PL dilauroylphosphatidylcholine (DLPC) which shows potent anti-diabetic effects. The discovery that LRH-1 is regulated by PL ligands reveals an exciting potential to tune LRH-1 activity for the treatment of metabolic diseases. However, PLs are labile and not suitable for clinical use, necessitating the development of small molecule agonists. This has proved challenging thus far, since very few small molecules can displace endogenous lipids from the large, lipophilic binding pocket. Recent studies in our lab have characterized a class of small molecules that are capable of this feat. We have designed potent LRH-1 agonists that display biological activity. We have modified our most potent and efficacious agonists to improve their biophysical properties, making them suitable for in vivo studies. The advancement of LRH-1 agonists as therapeutics has also been hindered by the lack of appropriate rodent models to screen potential candidates due to small sequence differences in the binding pocket of rodent and human LRH-1. To overcome this barrier, we used a CRISPR-Cas9 strategy to humanize the mouse LRH-1 ligand binding pocket. This permits activation by synthetic agonists while minimizing changes to endogenous interaction surfaces. These leaps forward in lead compound development and mouse model generation, in combination with our deep knowledge of LRH-1 structure and function, create an ideal platform to develop candidate preclinical LRH-1 modulators for metabolic disease. Here, we have developed a strategy to define mechanisms of action, target engagement, pharmacology, and disease efficacy of our lead compounds. In aim 1, we generate a compound with improved biophysical properties that mimics PL-like activation. We will perform mechanistic characterization of this compounds to explore how contacting the PL- binding site with different polar moieties improves LRH-1 activation. In aim 2, we will examine the behavior of our lead compound from an ADME perspective. The primary objective will be to establish tractability of the compounds using our humanized mice, so that pharmacokinetic relationships can be established. In aim 3, we will use our humanized mice and a model of diet-induced obesity to evaluate the in vivo efficacy of our lead LRH-1 compounds to improve glucose tolerance and insulin resistance.

肥胖在美国日益流行，导致非酒精性脂肪肝病 (NAFLD)、心血管疾病和 II 型糖尿病病例增加。这些疾病的共同特征是脂质和葡萄糖代谢异常。该提案以核激素受体、肝受体同源物 1 (LRH-1) 为中心，它是脂质代谢、逆转胆固醇转运、葡萄糖感应和体内平衡的重要调节剂。因此，LRH-1代表了代谢疾病的新治疗靶点。 LRH-1 与磷脂 (PL) 结合，并被不寻常的 PL 二月桂酰磷脂酰胆碱 (DLPC) 激活，显示出有效的抗糖尿病作用。 LRH-1 受 PL 配体调节的发现揭示了调节 LRH-1 活性以治疗代谢疾病的令人兴奋的潜力。然而，PL 不稳定，不适合临床使用，因此需要开发小分子激动剂。迄今为止，这已被证明具有挑战性，因为很少有小分子可以从大的亲脂性结合袋中取代内源性脂质。我们实验室最近的研究已经表征了一类能够实现这一壮举的小分子。我们设计了具有生物活性的强效 LRH-1 激动剂。我们修改了最有效的激动剂，以改善其生物物理特性，使它们适合体内研究。由于啮齿类动物和人类 LRH-1 的结合口袋存在微小的序列差异，缺乏合适的啮齿类动物模型来筛选潜在的候选药物，LRH-1 激动剂作为治疗药物的进展也受到阻碍。为了克服这一障碍，我们使用 CRISPR-Cas9 策略对小鼠 LRH-1 配体结合口袋进行人源化。这允许合成激动剂激活，同时最大限度地减少内源相互作用表面的变化。先导化合物开发和小鼠模型生成方面的这些飞跃，再加上我们对 LRH-1 结构和功能的深入了解，为开发用于代谢疾病的候选临床前 LRH-1 调节剂创造了一个理想的平台。在这里，我们制定了一项策略来定义我们的先导化合物的作用机制、靶点参与、药理学和疾病功效。在目标 1 中，我们生成了一种具有改善的生物物理特性的化合物，可模拟类 PL 激活。我们将对该化合物进行机械表征，以探索 PL 结合位点与不同极性部分的接触如何改善 LRH-1 激活。在目标 2 中，我们将从 ADME 角度检查先导化合物的行为。主要目标是使用我们的人源化小鼠确定化合物的易处理性，以便建立药代动力学关系。在目标 3 中，我们将使用人源化小鼠和饮食诱导的肥胖模型来评估我们的先导 LRH-1 化合物改善葡萄糖耐量和胰岛素抵抗的体内功效。