羟甲芬太尼对映异构体的镇痛活性及对阿片受体的选择性

羟甲芬太尼对映异构体的镇痛活性及对阿片受体的选择性金文桥，王智贤，陈洁，陈新建，池志强（中国科学院上海药物研究所，上海２０００３１，中国）关键词羟甲芬太尼；立体异构体；痛测定；μ阿片受体；δ阿片受体；放射配位体测定；结构活性关系目的：研究羟甲芬太尼...     

纳曲酮与纳洛酮对吗啡，依托尼秦及羟甲芬太尼的拮抗作用比较

在整体和受体水平对阿片受体拮抗剂纳曲酮和纳洛酮对吗啡，依托尼秦和［３Ｈ］羟甲芬太尼的拮抗作用进行了比较．研究表明，在整体水平，纳曲酮在很小剂量下就能对抗吗啡在小鼠的镇痛作用，小鼠吗啡急性中毒以及依托尼秦致大鼠翻正反射消失．与纳洛酮相比，纳曲酮强效，长效，ｉｇ有效．受体水平纳曲酮抑制［３Ｈ］羟甲芬太尼与大鼠脑阿片受体结合的强度是纳洛酮的３．６倍，与整体水平实验结果一致。     

Human μ-opioid receptor overexpressed in baculovirus system and its pharmacological characterizations

目的：高效表达具有类似哺乳动物特性的人μ阿片受体．方法：人μ阿片受体表达在重组杆状病毒感染的Ｓｆ９昆虫细胞中，用受体结合分析和ｃＡＭＰ分析研究表达产物的药理学特征．结果：［３Ｈ］二丙诺啡及［３Ｈ］羟甲芬太尼（Ｏｈｍ）的最大结合能力分别是９．１±０．７，６５２±０．２３ｎｍｏｌ／ｇ蛋白．μ选择性激动剂对［３Ｈ］二丙诺啡或［３Ｈ］Ｏｈｍ与表达受体的结合均有很强的抑制作用，而δ及κ激动剂则均无抑制作用．μ选择性激动剂有效抑制ｆｏｒｓｋｏｌｉｎ刺激的ｃＡＭＰ聚集，这种作用能被拮抗剂纳洛酮阻断．结论：在Ｓｆ９昆虫细胞中高效表达的人μ阿片受体保持着野生型人μ阿片受体的特征     

大鼠蛛网膜下腔联合注射kappa受体激动剂和NMDA受体拮抗剂协同镇痛

以大鼠热辐射甩尾潜伏期为测痛指标，蛛网膜下腔（ｉｔ）联合注射非镇痛剂量的ｋａｐｐａ阿片受体激动剂强啡肽（ｄｙｎｏｒｐｈｉｎ，Ｄｙｎ）Ａ－（１－１３）５ｎｍｏｌ或Ｕ５０４８８Ｈ（ｔｒａｎｓ－（±）－３，４－ｄｉｃｈｌｏｒｏ－Ｎ－ｍｅｔｈｙｌ－［２－（１－ｐｙｒｒｏｌｉｄｉｎｙｌ）－ｃｙｃｌｏｈｅｘｙｌ］－ｂｅｎｚｅｎｅａｃｅｔａｍｉｄｅ）１００ｎｍｏｌ和Ｎ－ｍｅｔｈｙｌ－Ｄ－ａｓｐａｒｔａｔｅ（ＮＭＤＡ）受体拮抗剂ＤＬ－２－ａｍｉｎｏ－５－ｐｈｏｓｐｈｏｎｏｖａｌｅｒｉｃａｃｉｄ（ＡＰｖ）１０ｎｍｏｌ或ｋｙｎｕｒｅｎｉｃａｃｉｄ（ＫＹＮ）５０ｎｍｏｌ有显著的协同镇痛效应，其效应与ＮＭＤＡ受体拮抗剂呈一定量效关系。Ｋａｐｐａ阿片受体特异性拮抗剂ｎｏｒ－ｂｉｎａｌｔｏｒｐｈｉ－ｍｉｎｅ（ｎｏｒ－ＢＮＩ）１５ｎｍｏｌｉｔ可完全翻转ＤｙｎＡ－（１－１３）５ｎｍｏｌ和ＡＰｖ１０ｎｍｏｌ及Ｕ５０４８８Ｈ１００ｎｍｏｌ和ＫＹＮ５０ｎｍｏｌ的协同镇痛。说明协同作用是通过ｋａｐｐａ受体和谷氨酸能神经元之间的相互作用实现的。     

Modulation of intrathecal morphine-induced immunosuppression by microinjection of naloxone into periaqueductal gray

目的：观察中脑导水管周围灰质（ＰＡＧ）的阿片受体和促皮质素（ＡＣＴＨ）在鞘内吗啡影响免疫功能时的作用和变化．方法：用铕标的靶细胞检测大鼠脾脏自然杀伤（ＮＫ）细胞活性，ＭＴＴ和结晶紫蓝法分析ＣｏｎＡ诱生的脾细胞白细胞介素２（ＩＬ２），肿瘤坏死因子ＴＮＦβ活性和血清ＴＮＦα水平，放免法测定血清促皮质素（ＡＣＴＨ）水平．结果：鞘内注射吗啡抑制脾ＮＫ细胞活性，ＣｏｎＡ诱生的ＩＬ２和ＴＮＦβ活性，并伴有血清ＡＣＴＨ水平的上升．ＰＡＧ微量注射纳洛酮部分拮抗吗啡引起的ＮＫ细胞活性的抑制，伴ＡＣＴＨ水平恢复．结论：ＰＡＧ内的阿片受体参与鞘内注射吗啡引起的ＮＫ细胞活性的抑制，同时伴有ＨＰＡ轴的激活     

新的高强度高选择性阿片μ受体激动剂［~11C］－羟甲芬太尼的合成

羟甲芬太尼（Ｉ）是一个新的高强度高选择性阿片μ受体激动剂。本文用ｃｉｓ－Ａ－Ｎ－［１－（２－羟基－２－苯乙基）－３－甲基－４－哌啶基］－苯胺（ＩＩ）或ｃｉｓ－Ｎ－［１－（苯甲酰甲基）－３－甲基－４－哌啶基］－苯胺（ＩＩＩ）作为前体合成了［１１Ｃ］－羟甲芬太尼，以便用正电子发射断层扫描（ＰＥＴ）来观察μ受体。通过水解ｃｉｓ－Ａ－羟甲芬太尼（Ｉ）和ｃｉｓ－Ｎ－［１－（苯甲酰甲基）－３－甲基－４－哌啶］－Ｎ－苯基丙酰胺（ｃｉｓ－ＩＶ）的４－Ｎ－丙酰基分别获得ＩＩ和ＩＩＩ。溴乙烷的格氏试剂与回旋加速器产生的［１１Ｃ］－二氧化碳反应后继而直接加入邻苯二甲酸二酰氯和２，６－二叔丁基吡啶生成同位素标记中间体［１１Ｃ］－丙酰氯。［１１Ｃ］－丙酰氯与ＯＨ－前体（ＩＩ）反应后再经ＨＰＬＣ分离纯化直接得［１１Ｃ］－羟甲芬太尼；［１１Ｃ］－丙酰氯与酮－前体（ＩＩＩ）反应后，再用硼氢化钠甲醇溶液处理，然后进行ＨＰＬＣ分离纯化得［１１Ｃ］－羟甲芬太尼。两种方法均可获得ｌｌ．１～１４．８ＧＢｑ／μｍｏｌ的特异性放射化学纯［１１Ｃ］－羟甲芬太尼。总共耗时为４０～５０ｍｉｎ（ＥＯＢ）。     

大鼠脑内不同部位注射降钙素对痛阈及Ca~（2＋）的影响

目的：观察降钙素对大鼠不同脑区痛阈的影响．方法：向大鼠不同脑区注射降钙素（ｃａｌｃｉｔｏｎｉｎ，Ｃａｌ），以辐射热甩尾阈值为指标测定痛阈变化．结果：Ｃａｌ侧脑室（ＶＬＣ）注射引起痛阈明显升高；同时发现向中脑导水管周围灰质（ｐｅｒｉａｑｕａｄｕｃｔａｌｇｒｅｙ，ＰＡＧ）注射Ｃａｌ，痛阈升高幅度更大，表明ＰＡＧ在Ｃａｌ的中枢镇痛效应中起重要作用．而向缰核（ｈａｂｅｎｕｌａ，Ｈａｂ）注射Ｃａｌ，痛阈降低．Ｃａｌ对痛阈的影响与脑内Ｃａ２＋有关．     

Antagonistic effect of orphanin FQ on opioid analgesia in rat

目的：研究孤啡肽（ＯＦＱ）对痛与阿片镇痛的影响．方法：脑室（ｉｃｖ）与鞘内（ｉｔｈ）给药，以大鼠甩尾模型测痛．结果：小剂量ＯＦＱ（０１μｇ）ｉｃｖ及ｉｔｈ给药对痛反应均无影响；较大剂量ＯＦＱ（０５－１０μｇ）可使痛反应增强．ＯＦＱ１－１０（ＯＦＱ的一个片段）ｉｃｖ对痛反应无影响．μ受体激动剂芬太尼（１μｇ）、δ激动剂ＤＳＬＥＴ（５μｇ）ｉｃｖ或ｉｔｈ给药，以及κ激动剂Ｕ５０４８８Ｈ（１μｇ）ｉｔｈ给药，可使痛阈明显增加．０１μｇ或１μｇＯＦＱ与上述药物合用后，痛阈增加明显减少（除鞘内与ＤＳＬＥＴ合用外）．结论：ＯＦＱ可增强大鼠的痛反应，在脑内对抗由μ和δ受体介导的阿片镇痛，在脊髓对抗由κ和μ但不是由δ受体介导的镇痛．     

羟甲芬太尼的精神依赖性系统评价

羟甲芬太尼是一种强效镇痛药，本实验采用３种动物、３种药物精神依赖性评价方法对其精神依赖性潜力进行系统评价，结果表明羟甲芬太尼存在着一定的精神依赖性潜力，但等效剂量的羟甲芬太尼的精神依赖性潜力低于吗啡，同时对其依赖性机理进行初步探讨推测其药物强化效应可能是通过阿片μ－受体起作用的。     

中药华蟾素的镇痛实验研究

目的探讨华蟾素注射液的镇痛作用及其机制。方法采用小鼠热板法、小鼠醋酸扭体法观察两种浓度华蟾素注射液的镇痛作用;采用盐酸纳洛酮实验研究华蟾素注射液镇痛作用机制。结果与氨酚羟考酮比较,华蟾素注射液能使醋酸所致小鼠扭体次数减少（P〈0．01）;并能提高热板所致小鼠舔足的痛阈（P〈0．05）：而且腹腔注射（IP）0．04mg纳洛酮能明显地拮抗华蟾素的镇痛作用（P〈0．001）。结论华蟾素具有镇痛作用,其镇痛作用可能与阿片受体有一定的关系。     

痛稳素碳末端八肽翻转孤啡肽对抗吗啡镇痛的作用

目的　观察痛稳素碳末端八肽在小鼠脑内对孤啡肽对抗吗啡的镇痛作用的影响。方法　固相多肽合成法合成了痛稳素碳末端八肽 ,用辐射热甩尾法测定痛阈 ,观察 (1)小鼠脑室注射 (icv)孤啡肽对吗啡镇痛作用的影响 ;(2 )小鼠脑室注射 (icv)痛稳素碳末端八肽对小鼠基础痛阈的影响 ;(3)小鼠脑室联合注射 (icv)痛稳素碳末端八肽和孤啡肽对吗啡镇痛作用的影响。结果　孤啡肽可对抗吗啡的镇痛作用 ;痛稳素碳末端八肽本身不影响小鼠的基础痛阈 ,但可逆转孤啡肽对抗吗啡的镇痛作用。结论　痛稳素碳末端八肽在脊髓以上水平可以逆转孤啡肽对抗吗啡的镇痛作用     

Studies on o-methylflavinantine

The antinociceptive effects of O-methylflavinantine (OMF), a morphinandienone alkaloid, were investigated in the mouse hot plate and abdominal constriction tests (nociceptive agents: 5-Hydroxytryptamine, acetylcholine, bradykinin, prostaglandin, E (1) (PGE (1), formic acid and phenylquinone). The potency of OMF in the hot plate test was approximately 10 times less than that of morphine and the effect was naloxone reversible. In the abdominal constriction test, morphine was 78-650 times more potent than OMF, depending on the nociceptive agent used, but a higher dose of naloxone was necessary to reverse the response to formic acid. Pretreatment of mice with reserpine (1 mg/kg, s.c., 24 h) reduced and alpha-methyl-p-tyrosine (200 mg/kg, i.p., 3 h) potentiated the antinociceptive effects of both morphine and OMF in the hot plate test. The results are considered to indicate that OMF possesses centrally mediated antinociceptive activity which is similar to that of morphine.     

孤啡肽与八肽胆囊收缩素在大鼠脑内拮抗吗啡镇痛的协同作用

目的观察孤啡肽（ＯＦＱ）和八肽胆囊收缩素（ＣＣＫ ８）在大鼠脑内拮抗吗啡镇痛是否具有协同作用。方法应用等高线法设计实验,用辐射热甩尾法测定痛阈。皮下注射吗啡（５ｍｇ·ｋｇ－１）２０ｍｉｎ之后,选择有明显镇痛效应的大鼠,侧脑室（ｉｃｖ）分别注射不同剂量的ＯＦＱ和ＣＣＫ ８以及由两者不同比例（５∶１,２５∶１）不同剂量组成的混合物,观察对吗啡镇痛效应的影响。结果联合应用ＯＦＱ和ＣＣＫ ８所产生的抗吗啡镇痛作用明显大于单独使用ＯＦＱ或ＣＣＫ ８。结论ＯＦＱ和ＣＣＫ ８在一定比例、一定剂量组合范围内,对抗吗啡镇痛具有协同效应。     

可乐定镇痛与中枢Ca2+的关系

用大鼠甩尾法和放射配基结合实验,探讨了可乐定镇痛与中枢Ca²⁺的关系。CaCl₂(1μmol/rat,icv)和EGTA(0.2μmol/rat,icv)分别拮抗和增强可乐定(1mg/kg,sc)的镇痛。戊脉安(0.1μmol/rat,icy)对可乐定(1 mg/kg,sc)镇痛无明显影响,但可部分翻转CaCl₂对可乐定镇痛的拮抗。CaCl₂(1×10^-3mol)对[³H]-可乐定结合无明显抑制。结果表明可乐定镇痛与脑室周围组织中Ca²⁺浓度变化密切相关,Ca²⁺至少部分需经对戊脉安敏感的钙通道进入细胞内方可拮抗可乐定镇痛。推沦:可乐定镇痛与神经元内Ca²⁺有关。     

Intrathecal N-methyl-D-aspartate (NMDA) induces paradoxical analgesia in the tail-flick test in rats

The intrathecal (IT) administration of NMDA in rodents has usually been reported to produce hyperalgesic reactions, although some articles describe that spinal NMDA can lead to analgesia. We show here that the nociceptive behavior (biting, scratching, licking; BSL) observed after NMDA injection (1-8 microg/rat; IT) is followed by a long period of increased tail-flick latencies, not longer detected 24 h after NMDA administration. The NMDA-receptor antagonist CPP (10-100 ng/rat; IT) blocked the BSL behavior induced by NMDA. In the tail-flick test, this antagonist induced analgesia by itself, and was able, at 30 ng/rat, to prevent the NMDA-mediated analgesia. The implication of opiate mechanisms was discarded since naloxone (3 and 10 mg/kg; IP) did not antagonize NMDA-induced analgesia. Finally, the involvement of the intracellular calcium binding protein calmodulin was assessed. The calmodulin inhibitor, calmidazolium (30-300 microg/rat; IT) only blocked the excitatory effect (BSL) without modifying the tail-flick analgesia produced by NMDA (4 microg). These results show that a single intrathecal administration of NMDA sequentially induces both nociceptive and antinociceptive, nonopiate responses in rats.     

Supraspinally mediated analgesic effect of antidepressant drugs

Imipramine and amitriptyline, nonselective monoamine reuptake inhibitors, citalopram, selective serotonin reuptake inhibitor, and maprotiline, selective noradrenaline reuptake inhibitor, were tested after intracerebroventricular (icv) and intrathecal (it) administration in the rat writhing test to establish the role of the spinal and/or supraspinal structures in their effects. All drugs evoked dose-dependent analgesia after icv but not after it microinjections. The changes in the nociceptive behavior of rats pre-treated with antidepressants were not due to the non-specific influences of the drugs on the animals' gross behavior, as revealed by the open field test. The obtained results suggest that an antidepressant-induced analgesia mainly depends on the supraspinal effect with minor, if any, contribution from the spinal mechanisms.     

Effect of intracerebroventricular injection of pentagastrin on rat drinking behavior

Intracerebroventricular (icv) injection of pentagastrin showed a powerful, dose dependent antidipsogenic effect in the rat. Drinking behavior stimulated by 48 h water deprivation was inhibited by 2000 ng of pentagastrin which also blocked, at lower doses, water intake induced by icv injection of angiotensin II (100 ng) and carbachol (150 ng). Pentagastrin was less effective on food intake stimulated by 24 h starvation. The antidipsogenic effect was not a consequence of behavioral alteration. It is suggested that gastrin-like peptides in the brain may play a role in the regulation drinking, acting as thirst inhibitors.