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.

项目总结 内源性大麻素2-花生烯酰-sn-甘油(2-AG)从突触后棘释放并激活 轴突终末上的CB1受体调节离子通道活动和神经递质释放。单甘酯 脂肪酶(MGL)是一种突触前水解酶，可以将2-AG水解成花生四烯酸和甘油，从而阻止这一过程 逆行信号传递过程。以MGL为靶点的小分子抑制剂提供了对 2-AG的功能，最近已进入临床开发阶段。当前的药物开发努力 在这一领域，重点是与MGL的催化部位不可逆地或可逆地相互作用的试剂。那里 然而，这两种方法都有严重的缺点：不可逆的抑制剂会导致过量的2- AG蓄积和随之而来的CB1脱敏，而可逆正构体抑制剂必须竞争 活性中心与2-AG和其他单酰甘油结合，可能达到较高的微摩尔浓度 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氢依赖 C201和C208在MGL中的亚磺化稳定了酶的催化失活构象，从而 增强CB1受体2-AG介导的信号转导。在这个修订的R21应用程序中，我们建议开发 靶向此调控区域的变构MGL抑制剂，通过这样做，可能能够绕过底物 竞争。特别是，我们鉴定了一类苯并异噻唑啉-3(2H)-酮衍生物，它能抑制MGL 通过以可逆的方式与调节半胱氨酸C201和C208相互作用而具有高效活性。 我们建议使用实验和计算相结合的方法--分子模拟， 构效关系研究、定点突变、质谱学和体内药理学 -修饰这些先导分子，产生有效的、具有系统活性的变构MGL抑制剂。这些 化合物将促进我们对2-AG介导的信号的理解，使我们能够测试2-AG 失活--这是内源性大麻素信号的核心--可能受生理因素的调节 以及影响这种状态的病理因素，如缺血。它们还可以作为 疼痛、物质使用障碍和其他人类疾病的新疗法的发现。