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

基本信息

批准号：
7575664
负责人：
Richard Irving Duclos
金额：
$ 19.5万
依托单位：
NORTHEASTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-01 至 2011-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7575664
关键词：
2-arachidonylglycerol Active Sites Affect Affinity Agonist Amides Anabolism Binding Biological Assay Blood - brain barrier anatomy Brain CNR1 gene Calcium ion Cannabinoids Carbamates Cell Communication Cells Characteristics Classification 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 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 Structure-Activity Relationship Substance abuse problem Symptoms System Thin Layer Chromatography Universities Wood material Work absorption addiction base cannabinoid receptor craving design drug candidate 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

项目摘要

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）：该提案的主要目的是了解二酰甘油甘油脂肪酶A（DAGL）在内生大麻素2-芳基二烯丙基甘油的生物合成中的作用，以及该酶在内ocnabanbinabinsible反应中的作用，与内ocnabinabinsisighotic反应相关的滥用效果，并具有相关性，并具有相关性，并具有相关性。第一个目标涉及从结构上衍生出DAGL底物（1,2-二酰基-SN-甘油）和选择性抑制剂四氢稳定蛋白（THL）的DAGL抑制剂的系统合成，该抑制剂的溶解度和吸收性差。所提出的结构特征包括：（a）针对DAGL酶的活性位点的缓慢可逆的反应性官能团（包括氨基甲酸酯，环甲氧甲氯苯甲酸，三氟甲基和B-内酰胺），以及（b）导致选择性结合到dagl的其他结构特征所必需的其他结构特征。第二个主要目的是开发用于DAGL抑制的高通量测定法，以取代当前使用的薄层色谱法。最后，可以通过分析其他脂肪酶活性的铅化合物来评估有效DAGL抑制剂的选择性。 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以及与大麻素受体CB1和CB2的亲和力。同样，由于静脉注射或口服剂量后，将评估其体内血浆和小鼠体内血浆中的DAGL抑制剂的潜在药物候选者，并使用定量质谱分析来评估其体内血浆和脑浓度，以筛选其在血液中分布在血脑屏障（BBB）上的能力（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。类似于缺乏特定的MAGL抑制剂，这代表了可剖析内源性大麻素系统的研究工具中的重要空隙，并研究了其与脂质代谢，钙离子传输，炎症，细胞信号传导和奖励大脑机制的微妙平衡。通过这种新型药物疗法降低2AG浓度应有效地拮抗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有关的医疗状况尤为重要，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了影响疼痛，饥饿和对滥用药物的渴望的细胞信号。许多当前流行的药物拮抗内源性大麻素受体蛋白，但是这种替代方法旨在通过选择性抑制其生物合成的方法来调节内差信号分子（2-蛛网膜烯烃）的水平。内源性大麻素信号的下调可能会对肥胖，成瘾中的恢复以及导致细胞通信途径变化的运动障碍症状产生有益的影响。