Selective inhibitors of ubiquitin E3 ligase to treat high cholesterol

泛素 E3 连接酶选择性抑制剂治疗高胆固醇

• 批准号: 8125555
• 负责人: David E Sterner
• 金额: $ 29.68万
• 依托单位: PROGENRA, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-19 至 2013-08-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8125555
• 关键词: Adult; Adverse effects; Adverse reactions; Affect; American; Binding; Biological Assay; Blood; Blood Circulation; Cardiovascular Diseases; Cell model; Cells; Chemicals; Cholesterol; Collection; Cytoplasmic Tail; Development; Disease; Dose; Drug Delivery Systems; Drug Industry; Dyslipidemias; Enzyme-Linked Immunosorbent Assay; Enzymes; Face; Family; Fingers; Goals; Heart Diseases; Hepatic; Human; Hydroxymethylglutaryl-CoA reductase; In Vitro; Individual; Laboratories; Lead; Liver; Low Density Lipoprotein Receptor; Low-Density Lipoproteins; Marketing; Modeling; Mutation; Nature; Nuclear; Pathway interactions; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Population; Proteins; Regulation; Reporting; Risk; Risk Factors; Screening procedure; Serum; Sterols; Therapeutic; Therapeutic Agents; Treatment Protocols; Ubiquitin; Validation; Work; base; cardiovascular disorder risk; cholesterol control; combinatorial; drug discovery; heart disease risk; high throughput screening; hypercholesterolemia; in vitro Assay; in vivo; inhibitor/antagonist; interest; multicatalytic endopeptidase complex; novel; pre-clinical; prevent; receptor; receptor upregulation; small molecule; ubiquitin-protein ligase; uptake

DESCRIPTION (provided by applicant): Hypercholesterolemia is a major factor in cardiovascular disease. In the USA, more than 35 million individuals have high total cholesterol and thus twice the normal risk of heart disease. The LDL receptor (LDLR) is critical to cholesterol regulation, as mutation of LDLR increases serum cholesterol levels and risk of cardiovascular disease. Increasing the level or activity of LDLR is an effective pharmacological mechanism employed by several cholesterol-lowering drugs now on the market. Statins, for example, inhibit HMG CoA reductase thus interfering with cholesterol synthesis, which upregulates hepatic LDLR activity and increases clearance of LDL from the bloodstream. Statins are widely prescribed, but are associated with adverse reactions in many patients; thus, new drugs of differing LDLR mechanism with reduced potential to cause side effects and/or potential for combination treatment employing reduced statin doses are currently being sought. An ubiquitin RING-finger E3 ligase known as Idol ( inducible degrader of the LDLR; also known as MYLIP and Mir), ubiquitylates LDLR on its cytoplasmic domain, resulting in its degradation. Idol is thus a promising new target for cholesterol-lowering drug discovery, as Idol inhibitors are expected to increase LDLR levels. Moreover, Idol inhibition provides a mechanistically distinct pathway to increase LDLR content/activity. The proposed project aims to discover small molecule inhibitors of Idol for lowering serum cholesterol by increasing LDLR levels and enhancing LDL clearance. Progenra has developed a novel assay that can detect the activity of virtually any E3 ligase; the assay has been used to find selective E3 ligase inhibitors, several of which are in pre-clinical development. Accordingly, the assay will be adapted and validated for high-throughput discovery of inhibitors of Idol, and a pilot screen will be conducted for inhibitors that are of potential utility as cholesterol- lowering therapeutic agents. The work will be divided into three Specific Aims. In the 1 year Phase I, the assay will be validated for inhibitors of both auto-ubiquitylation (Idol) and for inhibitors of ubiquitylation of the known substrate of human Idol. Cellular proof of concept will then be obtained experimentally using selected inhibitors from the screens. In Phase II, a more extensive screen and chemical optimization, ADME, and in vivo proof of concept and efficacy studies will be conducted, with the commercial goal of developing and marketing a pharmaceutical agent with single agent or combinatorial cholesterol lowering activity. PUBLIC HEALTH RELEVANCE: Elevated blood cholesterol is a major factor in cardiovascular disease; in the USA this condition affects more than 35 million individuals, who accordingly face increased risk of developing heart disease. The current drug of choice for lowering cholesterol is the family of statins, which work by increasing the binding of "bad" cholesterol to its receptors and thereby promote clearance of cholesterol from the bloodstream. Although widely prescribed, statins are associated with adverse reactions in many patients; thus, new drugs that would either work by a mechanism that leads to fewer or less severe side effects than those produced by the statins or be usable as part of a combination treatment regimen are currently being sought. Progenra has identified a promising new target for cholesterol-lowering drug discovery, an enzyme known as Idol, which conjugates ubiquitin to bad cholesterol receptors (LDLR), marking the receptor for degradation in the proteasome. Inhibitors of this target are expected to increase LDLR levels by preventing degradation of the receptor. Progenra will adapt and validate an assay for high throughput screening for inhibitors of Idol, screen its compound collection, and demonstrate that inhibitors from the screen affect LDLR in cells as predicted. In Phase II, selected inhibitors will undergo pre-clinical development, with the commercial goal of developing and marketing a pharmaceutical agent with single agent or combinatorial cholesterol lowering activity.

描述（由申请人提供）：高胆固醇血症是心血管疾病的主要因素。在美国，超过3500万人的总胆固醇高，因此是心脏病正常风险的两倍。随着LDLR的突变增加血清胆固醇水平和心血管疾病的风险，LDL受体（LDLR）对胆固醇的调节至关重要。 LDLR的水平或活性增加是一种有效的药理学机制，该机制现在在市场上使用了几种降低胆固醇的药物。他汀类药物，例如抑制HMG COA还原酶，因此会干扰胆固醇的合成，胆固醇的合成会上调肝LDLR活性并增加血液中LDL的清除率。他汀类药物被广泛处方，但与许多患者的不良反应有关。因此，目前正在寻求不同的LDLR机制的新药物，其潜力降低了潜在的副作用和/或潜在的结合治疗，目前正在寻求使用毒他汀类药物剂量减少的副作用。一个泛素环手指E3连接酶称为偶像（LDLR的诱导降解器；也称为mylip and mir），泛素化ldlr在其细胞质结构域上ldlr，从而导致其降解。因此，偶像是降低胆固醇药物发现的有希望的新靶标，因为偶像抑制剂有望提高LDLR水平。此外，偶像抑制提供了一种机械上不同的途径来增加LDLR含量/活动。拟议的项目旨在发现偶像的小分子抑制剂，用于通过提高LDLR水平并增强LDL清除率来降低血清胆固醇。后代已经开发了一种新颖的测定法，可以检测几乎任何E3连接酶的活性。该测定法已用于找到选择性的E3连接酶抑制剂，其中一些是在临床前发育中。因此，该测定法将进行调整和验证，以高通量发现偶像的抑制剂，并将为具有潜在效用的抑制剂进行试验筛查，以降低胆固醇的治疗剂。这项工作将分为三个特定目标。在第一阶段的一年中，该测定法将用于自动启发性（偶像）的抑制剂和人类偶像底物的泛素化抑制剂的抑制剂。然后，将使用筛选中选定的抑制剂实验获得概念证明。在第二阶段，将进行更广泛的屏幕和化学优化，ADME和体内概念和功效研究证明，其商业目标是开发和销售具有单药或组合胆固醇降低活性的药物。 公共卫生相关性：升高的血液胆固醇是心血管疾病的主要因素；在美国，这种情况会影响超过3500万人，因此他们面临着患心脏病的风险增加。当前降低胆固醇的首选药物是他汀类药物的家族，它们通过增加“不良”胆固醇与其受体的结合，从而促进血液中胆固醇清除。尽管处方广泛，但他汀类药物与许多患者的不良反应有关。因此，与他汀类药物产生的副作用相比，通过一种机制起作用的新药要么起作用，或者作为结合治疗方案的一部分可用。后代已经确定了降低胆固醇的药物发现的有希望的新靶标，该酶是一种称为偶像的酶，该酶将泛素结合到不良胆固醇受体（LDLR），标志着蛋白酶体中降解的受体。该靶标的抑制剂有望通过防止受体降解来提高LDLR水平。后代将适应和验证对偶像抑制剂的高吞吐量筛选的测定，其化合物收集，并证明筛网的抑制剂如预测的那样会影响细胞中的LDLR。在第二阶段，选定的抑制剂将经历临床前的开发，其商业目标是开发和销售具有单剂或组合胆固醇降低活动的药物剂。