SYNTHESIS, METABOLISM, AND PHARMACOLOGY OF ANANDAMIDE

阿南达胺的合成、代谢和药理学

基本信息

批准号：
2121327
负责人：
EARL F ELLIS
金额：
$ 18.55万
依托单位：
VIRGINIA COMMONWEALTH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1994
资助国家：
美国
起止时间：
1994-01-01 至 1996-12-31
项目状态：
已结题

项目摘要

In December, 1992 Devane et at. reported the discovery of an endogenous molecule isolated from brain that displace a cannabinoid receptor probe and possessed biologic activity similar to 9- tetrahydrocannabinol. (9-THC) The endogenous ligan was determined to be arachidonylethanolamide and was named anadamide (AN). We have obtained AN from Prof. Mechoulam and also synthesized our own radiolabeled and non-radiolabeled compound. Our preliminary experiments show that AN has 9-THC-like activity in that it produces analgesia, decreases spontaneous locomotion and produces hypothermia, Its CNS action appears to be weaker and of shorter duration than that of 9-THC. Our in vitro experiments suggest that AN can be synthesized by brain from H-arachidonic acid and that when added to brain AN is metabolized to rachidonic acid (AA). Arachidonic acid metabolism in brain is known to produce oxygen radicals and compounds which modulated brain blood flow. Our preliminary studies show that AN alone has little effect on cerebral blood vessels but its presence enhances the arteriolar response to acetylcholine and bradykinin, two agents known to activate prostaglandin formation. Therefore AN appears to affect behaviors as well as regulation of brain blood flow. We wish to test 3 hypotheses using techniques currently in our laboratories. Our first hypothesis is that AN produces the same pharmacological and behavioral profiles as 9-THC in mice and rats, as well as exhibiting similar receptor binding characteristics. This hypothesis will be examined using the tail flick assay for analgesia, studies of locomotion, drug discrimination and cannabinoid receptor binding. Or second hypothesis is that AN is synthesized and metabolized by brain and that factors which are known to release AA or alter AA metabolism affect AN formation. This hypothesis will be tested using radiolabeled probes, HPLC, gas chromatography/mass spectrometry, brain slices and primary cultures of neurons and astrocytes. Our third hypothesis is that metabolism of AN produces free AA which in turn affects cerebral blood flow regulation. This hypothesis will be tested using the closed cranial window technique for observation of in vivo cerebral arteriolar reactivity. Whether anandamide is the only or most active endogenous ligand for the cannabinoid receptor is uncertain. However the report of Devane et al. and our own preliminary studies support the likely importance of this newly discovered, biologically active molecule and emphasize the need for its rigorous examination. Our aims are consistent with our long term goals of elucidating cannabinoid pharmacology and the brain synthesis and action of arachidonic acid metabolites.

1992年12月，Devane et。报道了发现从大脑分离的内源分子，该分子位移大麻素受体探针和具有类似于9-的生物学活性四氢大麻酚。（9-thc）确定内源性凸是蛛网乙醇胺，被称为anadamide（AN）。我们从Mechoulam教授那里获得了一位，也合成了我们自己的放射性标记和非二醇标记化合物。我们的初步实验表明一个具有9-thc的活性产生镇痛，减少自发运动并产生体温过低，其中枢神经系统作用似乎较弱且较短持续时间比9-Thc的持续时间。我们的体外实验表明可以通过h-氨基苯二酚和当添加到大脑中时，将AN代谢为rachidonicac（AA）。已知大脑中的花生四烯酸代谢会产生氧气自由基和化合物调节脑血流。我们的初步研究表明，单独的对脑血管，但其存在增强了动脉对乙酰胆碱和心动激素的反应，两种已知的剂激活前列腺素形成。因此一个似乎影响行为以及脑血流的调节。我们希望使用目前在我们的技术中检验3个假设实验室。我们的第一个假设是一个产生相同的在小鼠和大鼠中，药理学和行为特征为9-THC，以及表现出相似的受体结合特征。该假设将使用尾部轻弹测定法进行镇痛，运动研究，药物歧视和大麻素受体结合。或第二个假设是由大脑合成和代谢，这些因素是已知释放AA或改变AA代谢会影响形成。该假设将使用放射性标记的探针HPLC，气体进行检验色谱/质谱法，大脑切片和原发性培养物神经元和星形胶质细胞。我们的第三个假设是代谢产生的自由AA，进而影响脑血流规定。该假设将使用封闭的颅骨进行检验观察体内脑小动脉的窗户技术反应性。 anandamide是唯一或最活跃的内源配体大麻素受体尚不确定。但是报告 Devane等。我们自己的初步研究支持这个新发现的生物活性分子的重要性并强调需要进行严格检查。我们的目标是与我们阐明大麻素的长期目标一致药理学与大脑合成和蛛网膜酸的作用代谢物。