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3500万人总胆固醇高，因此心脏病的风险是正常风险的两倍。LDL受体（LDLR）对胆固醇调节至关重要，因为LDLR的突变会增加血清胆固醇水平和心血管疾病的风险。增加LDLR的水平或活性是目前市场上几种降胆固醇药物采用的有效药理学机制。例如，他汀类药物抑制HMG CoA还原酶，从而干扰胆固醇合成，胆固醇合成上调肝脏LDLR活性并增加LDL从血流中的清除。他汀类药物被广泛处方，但在许多患者中与不良反应相关;因此，目前正在寻找具有不同LDLR机制的新药，其引起副作用的可能性降低和/或采用减少的他汀类药物剂量的联合治疗的可能性。被称为Idol的泛素环指E3连接酶（LDLR的诱导型降解剂;也称为MYLIP和Mir）使LDLR在其胞质结构域上泛素化，导致其降解。因此，Idol是降胆固醇药物发现的一个有前途的新靶点，因为Idol抑制剂有望增加LDLR水平。此外，Idol抑制提供了增加LDLR含量/活性的机制上不同的途径。该项目旨在发现Idol的小分子抑制剂，通过增加LDLR水平和增强LDL清除率来降低血清胆固醇。Progenra开发了一种新的检测方法，可以检测几乎任何E3连接酶的活性;该检测方法已被用于寻找选择性E3连接酶抑制剂，其中一些处于临床前开发阶段。因此，该测定法将被调整和验证用于Idol抑制剂的高通量发现，并且将对具有作为降胆固醇治疗剂的潜在效用的抑制剂进行中试筛选。这项工作将分为三个具体目标。在为期1年的I期研究中，将对自动泛素化（Idol）抑制剂和人Idol已知底物泛素化抑制剂的测定进行验证。然后将使用从筛选中选择的抑制剂通过实验获得概念的细胞证明。在II期，将进行更广泛的筛选和化学优化、ADME以及体内概念验证和疗效研究，商业目标是开发和销售具有单药或组合降胆固醇活性的药剂。 公共卫生相关性：血液胆固醇升高是心血管疾病的主要因素;在美国，这种情况影响超过3500万人，因此他们面临患心脏病的风险增加。目前选择用于降低胆固醇的药物是他汀类药物家族，其通过增加"坏"胆固醇与其受体的结合而起作用，从而促进血液中胆固醇的清除。虽然他汀类药物被广泛使用，但在许多患者中与不良反应有关;因此，目前正在寻求通过导致比他汀类药物产生的副作用更少或更不严重的副作用的机制起作用的新药，或者可用作联合治疗方案的一部分。Progenra已经确定了一个有希望的降胆固醇药物发现的新靶点，一种称为Idol的酶，它将泛素结合到坏胆固醇受体（LDLR）上，标记蛋白酶体中降解的受体。预期该靶点的抑制剂通过防止受体降解来增加LDLR水平。Progenra将调整并验证用于高通量筛选Idol抑制剂的试验，筛选其化合物集合，并证明筛选的抑制剂如预测的那样影响细胞中的LDLR。在II期，选定的抑制剂将进行临床前开发，商业目标是开发和销售具有单一药物或组合降胆固醇活性的药物。