内源性生物活性物质脂肪酰胺研究进展

生物体内存在着大量饱和或不饱和脂肪酰胺 ,它们自身的高生物活性、受体介导的作用机制以及特异的生物合成和降解途径 ,提示它们可能是一类新的生物调节分子家族。这一家族包括脂肪酸乙醇胺 ,如花生四烯酸乙醇胺(anandamide ,ANA)、棕榈酰乙醇胺 ( palmitoylethanolamine,PEA)等 ,和脂肪酸伯酰胺 ,如油酸酰胺 (oleamide,OLA )、芥子酸酰胺 (erucamide ,ERA)等。研究表明 ,内源性脂肪酰胺具有显著的中枢与外周活性。关于他们的生物学存在意义与作用机制的研究正在广泛深入地进行     

内源性睡眠促进物质研究进展

睡眠和觉醒的发生及转换可能是脑内神经递质和内源性睡眠促进物质共同作用的结果,同时受昼夜节律调控。Ishimori和Piéron首次发现犬睡眠剥夺后,脑脊液中某种化学物质增多,诱发睡眠,并将其命名为促眠素,即内源性睡眠促进物质。目前已发现前列腺素D2、核苷、细胞因子、神经肽、激素、胺类衍生物及一氧化氮等二十多种内源性睡眠促进物质。本文综述前列腺素D2、腺苷、白细胞介素1和肿瘤坏死因子等调节睡眠的作用机制研究进展。     

内源性大麻素样物质的可能作用机制

内源性大麻素样物质是大麻素受体的内源性配体 ,在脑内有其特定的合成、转运、代谢途径 ,在生理状态下可调节某些神经递质如谷氨酸、GABA等的释放。因此对其作用机制的阐明不仅有利于进一步探讨大麻成瘾的机制 ,还有助于新药的研发     

N-氨甲酰基-D-氨基酸酰胺水解酶研究进展

用生物转化法生产D－氨基酸已经成为国内外半合成抗生素行业的支柱之一。此外，D－氨基酸及其衍生物还是甜味剂、拟除虫菊脂、多肽激素等诸多产品的重要中间体。介绍D－氨基酸生物转化中N－氨甲酰基－D－氨基酸酰胺水解酶的一些生化性质、结构特征及催化机制等方面的研究进展。     

抑郁症患者外周血中大麻素受体1和脂肪酸酰胺水解酶的mRNA表达水平及意义

目的 探讨抑郁症患者外周血中大麻素受体1(CB1)和脂肪酸酰胺水解酶(FAAH)的mRNA表达水平及意义.方法 选择43例未经治疗的抑郁症患者(抑郁组)和43例健康志愿者(健康组),采用实时荧光定量反转录聚合酶链式反应(RT-PCR)技术检测两组患者外周血中CB1和FAAH的mRNA表达水平,观察抑郁症患者与健康人群的CB1和FAAH水平变化;采用汉密尔顿抑郁量表(HAMD)和Stroop色词测试对抑郁症患者的抑郁程度和认知功能进行评估,分析CB1和FAAH表达水平与测试结果的相关性.结果 (1)抑郁症患者的CB1-mRNA、FAAH-mRNA表达水平均明显低于健康组(P＜0.05);(2)抑郁症患者的CB1表达水平与HAMD中的体质量变化因子正相关(r=1.021,P=0.001);FAAH表达水平与HAMD中的认知障碍因子负相关(r=-0.065,P=0.023);(3)抑郁症患者FAAH-mRNA表达水平影响患者的注意力和反应能力(P＜0.05).结论 CB1受体可以影响抑郁症患者的能量代谢,FAAH会对抑郁症患者的认知障碍产生影响,提示抑郁症患者外周血CB1-mRNA、FAAH-mRNA表达量与抑郁障碍具有相关性.     

褪黑素与油酰胺联合应用后大鼠催眠作用的变化

目的研究两种内源性睡眠促进物质褪黑素和油酰胺联合应用后对大鼠催眠作用的变化。方法大鼠腹腔注射褪黑素(2.5,10mg·kg-1),油酰胺(25,50mg·kg-1)或联合应用褪黑素(2.5,10mg·kg-1)与油酰胺(25,50mg·kg-1)后,立即记录大鼠脑电和肌电变化。结果虽然褪黑素(2.5mg·kg-1)和油酰胺(25mg·kg-1)单独应用没有对大鼠脑电图产生明显影响,联合应用后显著缩短睡眠潜伏期(TSO)和觉醒时间(W),并且延长了慢波睡眠(SWS)和总睡眠时间(TS)。褪黑素(10mg·kg-1)和油酰胺(50mg·kg-1)单独具有明显的催眠作用,但联合应用后快波睡眠(PS)减少,其它睡眠指标未见变化。结论褪黑素和油酰胺联合应用后对大鼠催眠作用的变化较为复杂,其机制应进一步深入研究。     

双色胺—Nb,Nb′—二酰胺化合物的合成及其安定受体活性

本文由色胺或5－甲氧基色胺与相应的酰氯在三乙胺的催化下缩合合成了六个双色胺－Nb,Nb′－二酰胺化合物，其中四个为新化合物。经过放射配体结合竞争抑制性试验证明，该六个化合物均与安全受体有较好的亲和力。     

拉科酰胺原料药中有关物质的合成

目的研究拉科酰胺原料药中有关物质的来源及合成方法,为产品质量控制提供依据和杂质对照品。方法在参考拉科酰胺原料药相关合成方法的基础上,以Boc-D-丝氨酸为原料,经多步反应分别制得拉科酰胺的6种有关物质:(R)-2-氨基-N-苄基-3-羟基丙酰胺(A)、(R)-2-乙酰胺基-N-苄基-3-羟基丙酰胺(B)、(2R)-2-(乙酰胺基)-3-(乙酰氧基)-N-(苯甲基)丙酰胺(C)、N-[(1R)-1-(甲氧基甲基)-2-氧代-2-[(苯基甲基)氨基]乙基]-2-甲基丙酯(D)、(R)-N-苄基-2-(3-苄基脲基)-3-甲氧基丙酰胺(E)、(R)-N-苄基-3-甲氧基-2-(N-甲基乙酰氨基)丙酰胺(F)。结果与结论合成的6种拉科酰胺有关物质的结构经~1H-NMR、~(13)C-NMR、HR-MS谱确证,可作为拉科酰胺原料药的杂质对照品,其中有关物质E和F为新化合物。     

色胺Nb—酰胺衍生物的合成及其安定受体活性

报道了９个色胺－Ｎｂ－酰胺衍生物的合成，体外安定受体竞争结合试验初步结果表明，设计全成的化合物均有一定程度的安定受体亲和力，其中７个化合物的ＩＣ５０都小于Ｎ－（吲哚－３－基草酰基）甘氨酸乙酯。     

Systematic evaluation of a panel of 30 synthetic cannabinoid receptor agonists structurally related to MMB-4en-PICA,MDMB-4en-PINACA,ADB-4en-PINACA,and MMB-4CN-BUTINACA using a combination of binding and different CB1 receptor activation assays—Part II: Structure activity relationship assessment via a β-arrestin recruitment assay

Synthetic cannabinoid receptor agonists (SCRAs) are the second largest class of new psychoactive substances (NPS) and are associated with serious adverse effects and even death. Despite this, little pharmacological data are available for many of the most recent SCRAs. This study consists of three different parts, aiming to systematically evaluate a panel of 30 SCRAs using binding and different in vitro human cannabinoid 1 receptor (CB₁) activation assays. The present Part II investigated the SCRA analogs for their CB₁ activation via a β-arrestin recruitment assay. The panel was systematically designed to include key structural sub-features of recent SCRAs. Thus, the 4-pentenyl tail of MMB-4en-PICA and MDMB-4en-PINACA was retained while incorporating varying head groups from other prevalent SCRAs, including amides and esters of L-valine, L-tert-leucine, and L-phenylalanine, and adamantyl and cumyl moieties. All 30 SCRAs activated CB₁, with indazoles generally showing the greatest potency (EC₅₀ = 1.88–281 nM), followed by indoles (EC₅₀ = 11.5–2293 nM), and the corresponding 7-azaindoles (EC₅₀ = 62.4–9251 nM). Several subunit-linked structure–activity relationships were identified: (i) tert-leucine-functionalized SCRAs were more potent than the corresponding valine derivatives; (ii) no major difference in potency or efficacy was observed between tert-leucine/valine-derived amides and the corresponding methyl esters; however, phenylalanine analogs were affected by this change; and (iii) minor structural changes to the 4-pentenyl substituent had little influence on activity. These findings elucidate structural features that modulate the CB₁ activation potential of currently prevalent SCRAs and a systematic panel of analogs, some of which may appear in NPS markets in future.     

Evidence for both inverse agonism at the cannabinoid CB1 receptor and the lack of an endogenous cannabinoid tone in the rat and guinea-pig isolated ileum myenteric plexus-longitudinal muscle preparation

Background and purpose:

Cannabinoid receptor agonists reduce intestinal propulsion in rodents through the CB₁ receptor. In addition to its antagonistic activity at this receptor, rimonabant (N-(piperidino)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-3-pyrazole-carboxyamide) alone augments intestinal transit. Using rat and guinea-pig ileum MPLM (myenteric plexus-longitudinal muscle) preparations, we investigated whether the latter effect was through inverse agonism or antagonism of endocannabinoid agonist(s).

Experimental approach:

Inverse agonism was investigated by comparing the maximal enhancement of electrically evoked contractions of the MPLM by two CB₁ receptor antagonists, AM 251 (N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide) and O-2050 [(6aR,10aR)-3-(1-methanesulphonylamino-4-hexyn-6-yl)-6a,7,10,10a-tetrahydro-6,6,9-trimethyl-6-H-dibenzo[b,d]pyran], with that produced by rimonabant. To reveal ongoing endocannabinoid activity, effects of inhibiting endocannabinoid hydrolysis by fatty acid amide hydrolase (FAAH) using AA-5HT (arachidonyl-5-hydroxytryptamine), PMSF (phenylmethylsulphonyl fluoride) or URB-597 (3′-carbamoyl-biphenyl-3-yl-cyclohexylcarbamate), or putative uptake using VDM-11 [(5Z,8Z,11Z,14Z)-N-(4-hydroxy-2-methylphenyl)-5,8,11,14-eicosatetraenamide] was evaluated.

Key results:

The presence of CB₁ receptors was revealed by antagonism of exogenous anandamide, arachidonylethanolamide (AEA) and WIN 55,212-2 [(R)-(+)-[2,3-dihydro-5-methyl-3-(4-morpholinylmethyl)-pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-naphthalenylmethanone mesylate] by rimonabant. The rank order of potentiation of contractions was AM 251 > rimonabant > O-2050. Neither the FAAH inhibitors nor VDM-11 affected electrically evoked contractions. Each FAAH inhibitor increased the potency of AEA but not WIN 55,212-2. VDM-11 did not alter the inhibitory effect of AEA.

Conclusions and implications:

The different levels of maximal potentiation of contractions by the CB₁ receptor antagonists suggest inverse agonism. The potentiation of the action of AEA by the FAAH inhibitors showed that FAAH was present. The lack of effect of FAAH inhibitors and VDM-11 alone on electrically evoked contractions, and on the potency of exogenous AEA suggests that pharmacologically active endocannabinoids were not released and the endocannabinoid transporter was absent. Thus, the CB₁ receptor antagonists behave as inverse agonists.This article is part of a themed issue on Cannabinoids. To view the editorial for this themed issue visit http://dx.doi.org/10.1111/j.1476-5381.2010.00831.x     

Toxicological and structural features of organophosphorus and organosulfur cannabinoid CB1 receptor ligands.

Potent cannabinoid CB1 receptor ligands include anandamide [N-(2-hydroxyethyl)arachidonamide], Delta9-tetrahydrocannabinol, and 3H-CP 55,940 at the agonist site and selected organophosphorus esters (including some pesticides) and organosulfur compounds at a proposed closely coupled "nucleophilic" site. This study considers the toxicological and structural features of alkylfluorophosphonates, benzodioxaphosphorin oxides, alkanesulfonyl fluorides, and analogs acting at the nucleophilic site. Binding at the agonist site, using3H-CP 55,940 in assays with mouse brain membranes, is inhibited byO-isopropyl dodecylfluorophosphonate (compound 2), dodecanesulfonyl fluoride (compound 14) and dodecylbenzodioxaphosphorin oxide with IC50 values of 2-11 nM. Compounds 2 and 14 are also effectivein vivo, with 84% inhibition of mouse brain CB1 binding 4 h after intraperitoneal dosage at 30 mg/kg. Compound 14-inhibited CB1 in mouse brain requires about 3-4 days for recovery of 50% activity, suggesting covalent derivatization. Delayed toxicity (mortality in 0.3-5 days) from compounds 2, 14, and octanesulfonyl fluoride (18) is more closely associated with in vivo inhibition of brain neuropathy target esterase-lysophospholipase (NTE-LysoPLA) than with that of CB1 or acetylcholinesterase. NTE-LysoPLA inhibited by sulfonyl fluorides 14 and 18 cannot "age," a proposed requirement for NTE phosphorylated by organophosphorus-delayed neurotoxicants. Several octane- and dodecanesulfonamides with N-(2-hydroxyethyl) and other substituents based on anandamide give depressed mobility and recumbent posture in mice, but the effects do not correlate with potency for CB1 inhibition in vitro. Specific toxicological responses are not clearly associated with organophosphorus- or organosulfur-induced inhibition of the proposed CB1 nucleophilic site in mouse brain. On the other hand, the most potent CB1 inhibitors examined here are also NTE-LysoPLA inhibitors and cause delayed toxicity in mice.     

Involvement of 2-arachidonoyl glycerol in the increased consumption of and preference for ethanol of mice treated with neurotoxic doses of methamphetamine

Background and purpose:

Methamphetamine (METH) is a psychostimulant amphetamine that causes long-term dopaminergic neurotoxicity in mice. Hypodopaminergic states have been demonstrated to increase voluntary ethanol (EtOH) consumption and preference. In addition, the endocannabinoid system has been demonstrated to modulate EtOH drinking behaviour. Thus, we investigated EtOH consumption in METH-lesioned animals and the role of cannabinoid (CB) signalling in this EtOH drinking.

Experimental approach:

Mice were treated with a neurotoxic regimen of METH, and 7 days later exposed to increasing concentrations of drinking solutions of EtOH (3, 6, 10 and 20%). Seven days after neurotoxic METH, the following biochemical determinations were carried out in limbic forebrain: CB₁ receptor density and stimulated activity, 2-arachidonoyl glycerol (2-AG) and monoacylglycerol lipase (MAGL) activity, dopamine levels and dopamine transporter density.

Key results:

EtOH consumption and preference were increased in METH-treated mice. Seven days after METH, a time at which both dopamine levels and density of dopamine transporters in limbic forebrain were decreased, CB₁ receptor density and activity were unaltered, but 2-AG levels were increased. At this same time-point, MAGL activity was reduced. The CB₁ receptor antagonist AM251 prevented the METH-induced increase in EtOH consumption and preference, while N-arachidonoyl maleimide, an inhibitor of MAGL, increased EtOH consumption and preference in both saline- and METH-treated mice.

Conclusions and implications:

An increase in endocannabinoid tone may be involved in the increased consumption of and preference for EtOH displayed by METH-lesioned mice as blockade of the CB₁ receptor decreased EtOH-seeking behaviours, whereas the MAGL inhibitor increased EtOH consumption.This article is part of a themed issue on Cannabinoids. To view the editorial for this themed issue visit http://dx.doi.org/10.1111/j.1476-5381.2010.00831.x     

Systematic evaluation of a panel of 30 synthetic cannabinoid receptor agonists structurally related to MMB-4en-PICA,MDMB-4en-PINACA,ADB-4en-PINACA,and MMB-4CN-BUTINACA using a combination of binding and different CB1 receptor activation assays. Part III: The G protein pathway and critical comparison of different assays

The present work is the last of a three-part study investigating a panel of 30 systematically designed synthetic cannabinoid receptor agonists (SCRAs) including features such as the 4-pentenyl tail and varying head groups including amides and esters of l -valine (MMB, AB), l -tert-leucine (ADB), and l -phenylalanine (APP), as well as adamantyl (A) and cumyl moieties (CUMYL). Here, we evaluated these SCRAs for their capacity to activate the human cannabinoid receptor 1 (CB₁) via indirect measurement of G protein recruitment. Furthermore, we comparatively evaluated the results obtained from three in vitro assays, based on the recruitment of β-arrestin 2 (βarr2 assay) or Gα_i protein (mini-Gα_i assay), or binding of [³⁵S]-GTPγS. The observed efficacies (E_max) varied depending on the conducted assay. Statistical analysis suggests that the population means of the relative intrinsic activity (RA_i) significantly differ for the [³⁵S]-GTPγS assay and the other two assays, but the population means of the βarr2 and mini-Gα_i assays were not statistically different. Our data suggest that differences observed between the βarr2 and mini-Gα_i assays are the best predictor for ‘biased agonism’ towards βarr or G protein recruitment in our study. SCRAs carrying an ADB or MPP moiety as a head group tended to produce elevated E_max values in the βarr2 assay, which might result in a tendency of these compounds to cause pronounced tolerance in users—a hypothesis that should be evaluated further by future studies. In general, a comparison of efficacies derived from different assays is difficult and should only be conducted very cautiously.     

Genetic Deletion of Fatty Acid Amide Hydrolase Alters Emotional Behavior and Serotonergic Transmission in the Dorsal Raphe, Prefrontal Cortex, and Hippocampus

Pharmacological blockade of the anandamide-degrading enzyme, fatty acid amide hydrolase (FAAH), produces CB₁ receptor (CB₁R)-mediated analgesic, anxiolytic-like and antidepressant-like effects in murids. Using behavioral and electrophysiological approaches, we have characterized the emotional phenotype and serotonergic (5-HT) activity of mice lacking the FAAH gene in comparison to their wild type counterparts, and their response to a challenge of the CB₁R antagonist, rimonabant. FAAH null-mutant (FAAH^−/−) mice exhibited reduced immobility in the forced swim and tail suspension tests, predictive of antidepressant activity, which was attenuated by rimonabant. FAAH^−/− mice showed an increase in the duration of open arm visits in the elevated plus maze, and a decrease in thigmotaxis and an increase in exploratory rearing displayed in the open field, indicating anxiolytic-like effects that were reversed by rimonabant. Rimonabant also prolonged the initiation of feeding in the novelty-suppressed feeding test. Electrophysiological recordings revealed a marked 34.68% increase in dorsal raphe 5-HT neural firing that was reversed by rimonabant in a subset of neurons exhibiting high firing rates (33.15% mean decrease). The response of the prefrontocortical pyramidal cells to the 5-HT_2A/2C agonist (±)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ((±)-DOI) revealed desensitized 5-HT_2A/2C receptors, likely linked to the observed anxiolytic-like behaviors. The hippocampal pyramidal response to the 5-HT_1A antagonist, WAY-100635, indicates enhanced tonus on the hippocampal 5-HT_1A heteroreceptors, a hallmark of antidepressant-like action. Together, these results suggest that FAAH genetic deletion enhances anxiolytic-like and antidepressant-like effects, paralleled by altered 5-HT transmission and postsynaptic 5-HT_1A and 5-HT_2A/2C receptor function.     

Bioactivity-guided synthesis of gramine derivatives as new MT1 and 5-HT1A receptors agonists

Twenty-four gramine derivatives were synthesized and evaluated on MT₁ and 5-HT_1A receptors in vitro. Among them, seven derivatives (7, 8, 16, 19, 20, 21, and 24) exhibited higher agonisting activities on MT₁ or 5-HT_1A receptors. Compared with gramine, derivatives 7, 8, 16, 19, 20, 21, and 24 displayed 1.6–3.5-fold increase in agonistic rates on 5-HT_1A receptor. Particularly, derivatives 7, 19, and 21 exhibited significant agonistic activities on MT₁ and 5-HT_1A receptors with EC₅₀ values of 0.51, 0.39, 0.50 mΜ and 0.28, 0.46, 0.23 mΜ, respectively. The preliminary structure–activity relationships of gramine derivatives were summarized for further investigation on MT₁ and 5-HT_1A receptors as new potential agonists.     

孤啡肽研究进展

１９９５ 年底,发现了１ 种新型的内源性阿片肽———孤啡肽（ Ｎｏｃｉｃｅｐｔｉｏｎ 或 Ｏｒｐｈａｎｉｎ Ｆ Ｑ） ,它是１９９４ 年发现的阿片受体样（ Ｏ Ｒ Ｌ１ 或 Ｌ Ｃ１３２） 受体的内源性配体,它与受体结合后介导许多不同于经典阿片受体的生理活动。对孤啡肽的参与痛觉调节、神经传递、对离子通道和心血管系统的影响,以及拮抗剂与阻断剂、构效关系等方面的研究进展进行介绍