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: 10610353
• 负责人: Marco Mor
• 金额: $ 17.18万
• 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 2024-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10610353
• 关键词: 2-arachidonylglycerol; Active Sites; Affect; Arachidonic Acids; Area; Binding; Binding Sites; Brain; Bypass; CNR1 gene; Cannabis; Catalysis; Catalytic Domain; Central Nervous System; 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; Molecular; Molecular Conformation; Monoacylglycerol Lipases; Monoglycerides; 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; Shapes; Signal Transduction; Site; Site-Directed Mutagenesis; Spinal Cord; Structure-Activity Relationship; Substance Use Disorder; Sulfenic Acids; Sulfhydryl Compounds; Synapses; Synaptic Membranes; Testing; Tissues; Vertebral column; Work; alpha helix; 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; neural; neuropsychiatric disorder; neurotransmitter release; novel; novel therapeutics; oxidation; pharmacologic; postsynaptic; presynaptic; 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.

项目摘要 内源性大麻素2-蛛网膜烯酰基-SN-甘油（2-AG）从突触后棘中释放出来并激活 轴突末端上的CB1受体调节离子通道活性和神经递质释放。一甘油酸酯 脂肪酶（MGL） - 一种突触前水解酶，将2-AG水解为花生四烯酸和甘油 - 停止了这一点 逆行信号传导过程。靶向MGL的小分子抑制剂已提供了对 2-AG的功能最近已进入临床发展。目前的药物开发工作 在这一领域，集中于与MGL催化位点不可逆或可逆相互作用的药物。那里 但是，这两种方法都严重疾病：不可逆的抑制剂会导致过量2-- Ag积累和随之而来的CB1脱敏，而可逆的直角抑制剂必须竞争 与2-AG和其他单酰基甘油的活性位点结合，它们可能达到高度摩尔浓度 相关的生物相（例如，突触前膜）。我们已经表明，可逆的过氧化物依赖性 C201和C208在MGL中的磺苯基稳定酶的催化无效构象，从而稳定 增强在CB1受体处的2 AG介导的信号传导。在此修订后的R21应用程序中，我们建议开发 针对该调节区域的变构MGL抑制剂，并且通过这样做可能能够绕过底物 竞赛。特别是，我们鉴定了一类苯甲噻唑啉-3（2H） - 一种抑制MGL的衍生物 通过与调节性半胱氨酸C201和C208相互作用，具有高效力的活性。 我们建议使用实验和计算方法的组合 - 分子建模， 结构活性关系研究，定位的诱变，质谱和体内药理学 - 修改这些铅分子并产生潜力和系统活跃的变构抑制剂。这些 化合物将通过允许我们测试2-ag来提高我们对2-AG介导的信号传导的理解 停用（位于内源性大麻素信号的核心）可能会受到生理调节 以及影响这种状态的病理因素，例如缺血。他们也可能是起点 发现新颖的疼痛疗法，药物使用障碍和其他人类疾病。