Inhibition of Endocannabinoid Biosynthesis via Diacylglycerol Lipase

通过二酰基甘油脂肪酶抑制内源性大麻素生物合成

• 批准号: 7447608
• 负责人: Richard Irving Duclos
• 金额: $ 19.53万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7447608
• 关键词: 2-arachidonylglycerol; Active Sites; Affect; Affinity; Agonist; Amides; Anabolism; Binding; Biological Assay; Blood - brain barrier anatomy; Brain; CNR1 gene; CP-55,940; Calcium ion; Cannabinoids; Carbamates; Cell Communication; Cells; Characteristics; Classification; Condition; Data; Desire for food; Diacylglycerol Kinase; Dose; Down-Regulation; Endocannabinoids; Enzyme Inhibition; Enzymes; Equilibrium; Esters; Evaluation; Future; Glycerol; Goals; Health; Hunger; Hydrolase; In Vitro; Inflammation; Inhibitory Concentration 50; Intravenous; Ion Transport; Lactams; Lead; Letters; Ligands; Lipase; Medical; Monoacylglycerol Lipases; Movement Disorders; Mus; Numbers; Obesity; Oral; Pain; Pancreas; Pathway interactions; Pharmaceutical Preparations; Pharmacotherapy; Physiological; Plasma; Preparation; Property; Proteins; Rattus; Reader; Recovery; Research; Rewards; Role; Signal Transduction; Signaling Molecule; Solubility; Specificity; Standards of Weights and Measures; Structure-Activity Relationship; Substance abuse problem; Symptoms; System; Thin Layer Chromatography; Universities; Urination; Wood material; Work; absorption; addiction; base; cannabinoid receptor; craving; design; drug discovery; drug of abuse; fatty acid amide hydrolase; functional group; improved; in vivo; inhibitor/antagonist; lipid metabolism; lipoprotein lipase; novel; orlistat; receptor; research study; response; reuptake; tool

DESCRIPTION (provided by applicant): The primary goal of this proposal is an understanding of the role of diacyglycerol lipase-a (DAGL) in the biosynthesis of the endogeneous cannabinoid 2-arachidonoylglycerol and the role of this enzyme in endocannabinoid signaling and physiological responses related to appetite, substance abuse, and related health effects. The first objective involves a systematic synthesis of DAGL inhibitors structurally derived from DAGL substrate (1,2-diacyl-sn-glycerol) and the selective inhibitor tetrahydrolipstatin (THL), which has poor solubility and absorption characteristics. The proposed structural features include: (a) slowly reversible reactive functional groups (including carbamate, cyclohexyloximinocarbonylamino, trifluoromethyl, and b-lactam) targeted at the active site of DAGL enzyme, and (b) other structural characteristics necessary to result in selective binding to DAGL. The second major objective is to develop a highthroughput assay for DAGL inhibition, to replace the thin layer chromatography assay that is currently used. Finally, the selectivity of potent DAGL inhibitors can then be evaluated by assaying lead compounds for other lipase activities. These newly synthesized DAGL inhibitors will be assayed for inhibition of pancreatic lipase (TAGL), acetylcholineesterase (AChE), diacylglycerol kinase, and the endocannabinoid proteins: monoacylglycerol lipase (MAGL), fatty acid amide hydrolase (FAAH), the putative endocannabinoid transporter system, as well as for affinity to the cannabinoid receptors CB1 and CB2. Also, potential drug candidates with greater than fivefold selectivity as DAGL inhibitors will be evaluated for their in vivo blood plasma and brain concentrations in mouse, following intravenous or oral dosing, utilizing quantitative mass spectrometric analysis to screen for their ability to distribute across the blood-brain barrier (BBB). There are no highly specific DAGL inhibitors having appropriate solubility, absorption, and distribution properties. Analogously to the lack of specific MAGL inhibitors, this represents a significant void in the research tools available to dissect the endocannabinoid system and study its delicate balance with lipid metabolism, calcium ion transport, inflammation, cell signaling, and reward mechanisms in the brain. Lowering 2-AG concentrations by this novel pharmacotherapy should effectively antagonize CB receptors and affect endocannabinoid signaling, as well as CB receptor reuptake, plasticity, and crosstalk. The downregulation of the endocannabinoid system is particularly important for medical conditions related to overstimulation of the cannabinoid receptor including recovery from substance abuse, obesity, and movement disorders. The endocannabinoid system is involved in cell signalings which affect pain, hunger, and cravings for drugs of abuse. A number of currently popular medications antagonize the endocannabinoid receptor proteins, but this proposed alternative approach seeks to modulate the levels of the endogeneous signaling molecule (2- arachidonoylglycerol) by selectively inhibiting its biosynthesis. This downregulation of endocannabinoid signaling may have a benefical effect on obesity, recovery from addiction, and movement disorder symptoms from the resulting changes in cell communication pathways.

描述（由申请人提供）：本提案的主要目标是了解二酰甘油脂肪酶-a (DAGL) 在内源性大麻素 2-花生四烯酰甘油生物合成中的作用，以及该酶在内源性大麻素信号传导和相关生理反应中的作用食欲、药物滥用和相关的健康影响。第一个目标涉及系统合成结构上源自 DAGL 底物（1,2-二酰基-sn-甘油）的 DAGL 抑制剂和选择性抑制剂四氢利普他汀（THL），其溶解度和吸收特性较差。拟议的结构特征包括：（a）针对 DAGL 酶活性位点的缓慢可逆反应官能团（包括氨基甲酸酯、环己基肟基氨基、三氟甲基和 β-内酰胺），以及（b）导致选择性结合的其他结构特征达格。第二个主要目标是开发一种 DAGL 抑制的高通量测定法，以取代目前使用的薄层色谱测定法。最后，可以通过测定先导化合物的其他脂肪酶活性来评估有效 DAGL 抑制剂的选择性。这些新合成的 DAGL 抑制剂将用于抑制胰腺脂肪酶 (TAGL)、乙酰胆碱酯酶 (AChE)、二酰甘油激酶和内源性大麻素蛋白：单酰基甘油脂肪酶 (MAGL)、脂肪酸酰胺水解酶 (FAAH)、推定的内源性大麻素转运系统、以及对大麻素受体 CB1 和 CB2 的亲和力。此外，作为 DAGL 抑制剂，具有超过五倍选择性的潜在候选药物将在静脉内或口服给药后评估其在小鼠体内血浆和脑中的浓度，利用定量质谱分析来筛选它们在血液中分布的能力。脑屏障（BBB）。没有具有适当溶解度、吸收和分布特性的高度特异性 DAGL 抑制剂。与缺乏特定的 MAGL 抑制剂类似，这代表了可用于剖析内源性大麻素系统并研究其与脂质代谢、钙离子转运、炎症、细胞信号传导和大脑奖励机制之间的微妙平衡的研究工具的重大空白。通过这种新型药物疗法降低 2-AG 浓度应能有效拮抗 CB 受体并影响内源性大麻素信号传导，以及 CB 受体的再摄取、可塑性和串扰。内源性大麻素系统的下调对于与大麻素受体过度刺激相关的医疗状况尤其重要，包括从药物滥用、肥胖和运动障碍中恢复。内源性大麻素系统参与影响疼痛、饥饿和对滥用药物的渴望的细胞信号传导。目前流行的许多药物都会拮抗内源性大麻素受体蛋白，但这种提议的替代方法旨在通过选择性抑制内源性信号分子（2-花生四烯酰甘油）的生物合成来调节其水平。内源性大麻素信号传导的下调可能对肥胖、成瘾恢复以及细胞通讯途径变化产生的运动障碍症状产生有益的影响。