Allosteric control of monoacylglyceride lipase (MGL) activity

单酰甘油脂肪酶 (MGL) 活性的变构控制

• 批准号: 10373841
• 负责人: Marco Mor
• 金额: $ 22.53万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373841
• 关键词: 2-arachidonylglycerol; 2-arachidonylglycerol signaling; Active Sites; Affect; Arachidonic Acids; Area; Binding; Binding Sites; Brain; Bypass; CNR1 gene; Cannabis; Catalysis; Catalytic Domain; Chemical Agents; Chemicals; Cysteine; Disadvantaged; Disease; Drug Addiction; Endocannabinoids; Enzyme Inhibitor Drugs; Enzymes; Family; Feasibility Studies; Frequencies; Gilles de la Tourette syndrome; Glycerol; Goals; Human; Hydrogen Peroxide; Hydrolase; Ion Channel; Ischemia; Lead; Ligands; Lipase; Mass Spectrum Analysis; Mediating; Membrane; Molecular; Molecular Conformation; Monoacylglycerol Lipases; Monoglycerides; Neuraxis; Neurons; Neurotransmitters; Nucleic Acid Regulatory Sequences; Oxidation-Reduction; Pain; Pain management; Pathologic; Pathology; Peripheral; Peroxides; Pharmacology; Physiological; Positioning Attribute; Post-Translational Protein Processing; Post-Traumatic Stress Disorders; Presynaptic Terminals; Process; Research; Resolution; Second Messenger Systems; Signal Transduction; Site; Site-Directed Mutagenesis; Spinal Cord; Structure-Activity Relationship; Substance Use Disorder; Sulfenic Acids; Sulfhydryl Compounds; Synapses; Testing; Tissues; Vertebral column; Work; alpha helix; base; beta pleated sheet; cannabinoid receptor; clinical development; desensitization; drug development; drug discovery; endocannabinoid signaling; experience; human disease; in vitro activity; in vivo; in vivo evaluation; inhibitor; innovation; insight; molecular modeling; neuropsychiatric disorder; neurotransmitter release; novel; novel therapeutics; oxidation; postsynaptic; presynaptic; relating to nervous system; small molecule inhibitor; therapeutic target; therapy development

PROJECT SUMMARY The endocannabinoid 2-arachidonoyl-sn-glycerol (2-AG) is released from postsynaptic spines and activates CB1 receptors on axon terminals to regulate ion-channel activity and neurotransmitter release. Monoglyceride lipase (MGL) – a presynaptic hydrolase that hydrolyzes 2-AG into arachidonic acid and glycerol – stops this retrograde signaling process. Small-molecule inhibitors that target MGL have provided insights into the functions of 2-AG and have been recently advanced to clinical development. Current drug development efforts in this area are focused on agents that interact irreversibly or reversibly with the catalytic site of MGL. There are, however, serious disadvantages to either of these approaches: irreversible inhibitors cause excessive 2- AG accumulation and consequent CB1 desensitization, while reversible orthosteric inhibitors must compete for active-site binding with 2-AG and other monoacylglycerols, which may reach high micromolar concentrations in the relevant biophase (e.g., presynaptic membranes). We have shown that the reversible peroxide-dependent sulfenylation of C201 and C208 in MGL stabilizes a catalytically inactive conformation of the enzyme, and thus enhances 2-AG-mediated signaling at CB1 receptors. In this revised R21 application, we propose to develop allosteric MGL inhibitors that target this regulatory region and, by doing so, may be able to bypass substrate competition. In particular, we identified a class of benzisothiazolin-3(2H)-one derivatives that inhibit MGL activity with high potency by interacting in a reversible manner with the regulatory cysteines C201 and C208. We propose to use a combination of experimental and computational approaches – molecular modeling, structure-activity relationship studies, site-directed mutagenesis, mass spectrometry, and in vivo pharmacology – to modify these lead molecules and generate potent and systemically active allosteric MGL inhibitors. These compounds will advance our understanding of 2-AG-mediated signaling by allowing us to test whether 2-AG deactivation – which lies at the very core of endocannabinoid signaling – might be modulated by physiological and pathological factors that affect such status, such as ischemia. They might also serve as starting point for the discovery of novel therapeutics for pain, substance use disorders and other human diseases.

项目概要 内源性大麻素 2-花生四烯酰-sn-甘油 (2-AG) 从突触后棘释放并激活 轴突末端的 CB1 受体调节离子通道活动和神经递质释放。 脂肪酶 (MGL)——一种突触前水解酶，可将 2-AG 水解成花生四烯酸和甘油——可以阻止这种情况发生 针对 MGL 的逆行信号传导过程提供了深入的见解。 2-AG 的功能，最近已进入临床开发工作。 该领域的研究重点是与 MGL 催化位点发生不可逆或可逆相互作用的试剂。 然而，这两种方法都有严重的缺点：不可逆抑制剂会导致过量的 2- AG 积累和随后的 CB1 脱敏，而可逆的正位抑制剂必须竞争 与 2-AG 和其他单酰基甘油的活性位点结合，可能达到高微摩尔浓度 相关的生物相（例如，突触前膜）我们已经证明了可逆的过氧化物依赖性。 MGL 中 C201 和 C208 的磺酰化稳定了酶的催化失活构象，因此 增强 2-AG 介导的 CB1 受体信号传导 在这个修订后的 R21 应用中，我们建议开发。 靶向该调节区域的变构 MGL 抑制剂可能能够绕过底物 特别是，我们鉴定了一类抑制 MGL 的 Benzisothiazolin-3(2H)-one 衍生物。 通过与调节性半胱氨酸 C201 和 C208 以可逆方式相互作用，具有高效活性。 我们建议结合使用实验和计算方法——分子建模， 构效关系研究、定点诱变、质谱分析和体内药理学 – 修饰这些先导分子并产生有效且具有系统活性的变构 MGL 抑制剂。 化合物将通过允许我们测试 2-AG 是否 失活——这是内源性大麻素信号传导的核心——可能受到生理调节 影响这种状态的病理因素，例如缺血，它们也可能作为起点。 发现治疗疼痛、药物滥用障碍和其他人类疾病的新疗法。