四氢原小檗碱先导化合物l-SLR抗精神分裂症的药效学和机制研究

发布时间：2018-03-16 02:14

本文选题：精神分裂症　切入点：左旋金黄紫堇碱　出处：《中国人民解放军军事医学科学院》2016年硕士论文　论文类型：学位论文

【摘要】：精神分裂症是一种严重的脑功能障碍疾病,临床上根据症状表现一般可分为阳性症状(妄想、幻觉和幻听)和阴性症状(社会功能退缩等),以及以注意力、执行能力、解决问题能力和短期记忆能力缺失或减弱等为主的认知障碍。目前临床上的抗精神病药物均能拮抗多巴胺D2受体,经典抗精神病药物氟哌啶醇对阳性症状有效,但对阴性症状和认知障碍无效甚至加重;精神分裂症的阴性症状和认知障碍难于治疗,有些病人即使长期服药认知障碍也有可能持续存在,因此改善病人阴性症状与认知障碍是治疗精神分裂症的核心问题。为了开发对精神分裂症的阴性症状和认知障碍均有效的新型药物,我们进行了如下研究:实验目的:评价化合物左旋金黄紫堇碱(l-Scoulerine,l-SLR)对精神分裂症阳性症状、阴性症状以及认知障碍的作用,并运用放射性配体竞争性拮抗实验对其机制进行研究,旨在开发对精神分裂症的阳性症状、阴性症状以及认知障碍均有效且锥体外系不良反应小或无的新型抗精神病药。实验方法:采用自发活动实验测定化合物l-SLR是否具有镇静作用以及镇静作用持续时间。NMDA受体拮抗剂MK-801可引发精神分裂症的各种临床症状。采用前脉冲抑制实验和攀爬实验来评价化合物l-SLR对精神分裂症阳性症状的作用。在前脉冲抑制实验中,以大鼠腹腔注射MK-801(0.3 mg/kg)引起的大鼠前脉冲抑制缺失进行建模,观察大鼠在注射化合物l-SLR(5,10,15 mg/kg)之后能否逆转这种前脉冲抑制损伤,并将化合物l-SLR与典型抗精神病药氟哌啶醇(0.2mg/kg)的作用进行比较;在攀爬实验中,以小鼠皮下注射阿扑吗啡(2 mg/kg)引起小鼠攀爬行为进行建模,观察小鼠在腹腔注射化合物l-SLR(10,30 mg/kg)之后对小鼠攀爬时间的影响,并将此作用与氟哌啶醇进行比较。在精神分裂症的阴性症状中,采用小鼠的社交回避实验对化合物l-SLR进行药效学评价,在此实验中,以给小鼠腹腔注射工具药MK-801(0.2 mg/kg),使陌生的小鼠出现社交抑制进行建模,测定小鼠在注射化合物l-SLR之后能否增加陌生成对小鼠的接触时间,并比较化合物l-SLR和典型抗精神病药氟哌啶醇对精神分裂症阴性症状的作用。在精神分裂症的认知障碍中,采用y迷宫实验验证化合物l-slr抗精神分裂症的作用,以小鼠在注射mk-801(0.2mg/kg)后出现空间分辨能力降低,即精神分裂症的认知障碍进行建模,观察注射化合物l-slr能否提高小鼠的空间认知能力,并将此作用与经典的抗精神病药氟哌啶醇进行比较。在锥体外系不良反应方面,采用化合物l-slr与典型的抗精神病药氟哌啶醇和类似物l-spd进行比较,测定小鼠在注射氟哌啶醇(0.8mg/kg)、l-slr(10,30,60,80mg/kg)和l-spd(30,60,80mg/kg)之后的45和90分钟后的木僵时间,比较不同化合物之间对锥体外系的不良反应。在机制研究中,采用放射性生物配体的竞争性抑制测定化合物l-slr与多巴胺受体亲和力。实验结果:在自发活动实验中,小鼠在注射l-slr后的前30分钟内活动性降低,而在之后的时间里活动性恢复,说明化合物l-slr具有镇静作用,此作用的持续时间为30分钟,在实验设计过程中要避免在此段时间内进行药效学研究。在前脉冲抑制实验中,与veh组相比,mk-801(0.3mg/kg)能够明显降低大鼠的前脉冲抑制中的震惊反射(f5,44=6.696,p0.0001),而注射l-slr(10mg/kg、15mg/kg)和氟哌啶醇(0.2mg/kg)能够明显改善由mk-801引起的前脉冲抑制损伤(p0.01),这说明化合物l-slr和氟哌啶醇对mk-801引起的前脉冲抑制损伤均有效;在攀爬实验中,与veh组相比,阿扑吗啡(2mg/kg)皮下注射能够明显增加小鼠的攀爬时间(f4,45=19.13,p0.0001),氟哌啶醇(0.2mg/kg)和l-slr(30mg/kg)能够明显抑制阿扑吗啡所引起的攀爬行为,这说明化合物l-slr和氟哌啶醇对阿扑吗啡引起的小鼠攀爬行为有效。在小鼠的群居接触实验中,与veh组相比,mk-801能够明显抑制小鼠的群居接触行为,使小鼠的群居接触时间明显降低(f3,36=7.566,p=0.0084),而典型的抗精神病药氟哌啶醇却不能使小鼠的群居接触时间延长;与之相对应,化合物l-slr在30mg/kg时能够明显增加小鼠的群居接触时间(f5,54=6.285,p=0.0001),说明l-slr在剂量为30mg/kg时对精神分裂症的阴性症状有效,而氟哌啶醇无效。在精神分裂症认知障碍的y迷宫中,与veh组相比,mk-801能够明显地抑制小鼠的空间认知能力(f3,36=10.60,p0.0001),给予氟哌啶醇并不能改善小鼠的这种空间认知损伤;而化合物l-slr在剂量为30mg/kg时能够明显改善小鼠的空间认知障碍(f5,54=11.58,p0.0001),说明l-slr在剂量为30mg/kg时能够改善精神分裂症的认知损伤。在锥体外系不良反应方面,与veh组相比,在给药45分钟后,氟哌啶醇(0.8mg/kg)和l-spd(60mg/kg、80mg/kg)能够使小鼠的木僵时间明显增加(f8,64=13.63,p0.001);给药90分钟以后,氟哌啶醇和l-SPD(30mg/kg、60 mg/kg、80 mg/kg)引起的木僵行为仍然存在(F8,64=14.54,P0.001),而l-SLR在30mg/kg、60 mg/kg并不产生木僵,说明l-SLR在抗精神分裂症的有效剂量下不产生锥体外系的副作用。在化合物l-SLR的机制研究中,18F-Fallypride在进入小鼠体内有后迅速分布进入小鼠大脑的各个部分,而在给予18F-Fallypride之后迅速给予l-SLR的小鼠中,18F-Fallypride在大脑各个部位的分布在给药15分钟以后均明显低于FP组(P0.05),这说明化合物l-SLR能够与脑内的D2与D3受体相结合,从而能够拮抗18F-Fallypride与之的结合。结论:从自发活动实验可以看出,化合物l-SLR具有镇静作用,而且镇静时间较短,因此我们在对化合物的药效学研究中要避免在此段时间内进行。从前脉冲抑制实验和攀爬实验可以看出,化合物l-SLR能够对精神分裂症的阳性症状的结论。从社交回避的实验中,化合物l-SLR对精神分裂症的阴性症状有效。从Y迷宫实验可以看出,化合物l-SLR能够改善精神分裂症的认知障碍。因此,在治疗精神分裂症的认知障碍方面,l-SLR优于氟哌啶醇。在锥体外系不良反应方面,化合物l-SLR在有效剂量时不会引起小鼠的木僵,说明化合物l-SLR较少地引起小鼠锥体外系不良反应,相对于氟哌啶醇和l-SPD有显著的优势。在受体结合的研究中,通过拮抗18F-Fallypride使各个脑区的放射性降低,说明化合物l-SLR能够与多巴胺D2、D3受体结合。
[Abstract]:Schizophrenia is a severe brain dysfunction disease, according to clinical symptoms in general can be divided into positive symptoms (hallucinations and delusions, hallucinations) and negative symptoms (such as social function, and attention to withdrawal), executive ability, cognitive impairment based problem-solving ability and short-term memory capacity is absent or reduced. At present the Clinical Antipsychotic drugs can antagonize dopamine D2 receptor, antipsychotic drugs haloperidol effective on positive symptoms, negative symptoms and cognitive impairment but ineffective or aggravated; negative symptoms and cognitive impairment in schizophrenia is difficult to treatment, some patients even long-term medication of cognitive impairment may also continue to exist, so improving patients with negative symptoms and cognitive impairment is the key problem in the treatment of schizophrenia. In order to develop a model of schizophrenia with negative symptoms and cognitive impairment. Drug, we carry out the following research: Objective: To evaluate compound L - scoulerine (l-Scoulerine, l-SLR) of the positive symptoms of schizophrenia negative symptoms and cognitive impairment, and using radioligand competitive antagonism experiment to study its mechanism, aims to develop the positive symptoms of schizophrenia, and negative symptoms cognitive disorders are effective and extrapyramidal antipsychotic drugs have little or no adverse reactions. Methods: the spontaneous activity test compounds was determined whether l-SLR has a sedative effect and sedation for various clinical symptoms can cause schizophrenia with duration of.NMDA receptor antagonist MK-801. The pre pulse inhibition and climbing experiment evaluation of compound l-SLR on the positive symptoms of schizophrenia. The role in prepulse inhibition in rats by intraperitoneal injection of MK-801 (0.3 mg/kg) led The rats of prepulse inhibition were lack of modeling, rats in injection of compound l-SLR (5,10,15 mg/kg) can reverse the prepulse inhibition after injury, and the compound l-SLR with typical antipsychotic haloperidol (0.2mg/kg) were compared in the climbing effect; in the experiment, the mice subcutaneous injection of apomorphine (2 mg/kg) climbing behavior caused by modeling, were observed in mice after intraperitoneal injection of compound l-SLR (10,30 mg/kg) after impact on the climbing time of the mice, and the effects were compared with haloperidol. The negative symptoms of schizophrenia, the mice experiment on social avoidance efficacy evaluation of compound l-SLR, in this experiment, in order to give medicine intraperitoneal injection of MK-801 (0.2 mg/kg), the mice appeared strange social inhibition model of mice was measured after injection of compound l-SLR can increase into strange The contact time of mice, and compounds l-SLR and typical antipsychotics haloperidol on negative symptoms of schizophrenia. The cognitive deficits in schizophrenia, using Y maze experiment of compound l-slr anti schizophrenia effects in mice by injection of MK-801 (0.2mg/kg) after lower spatial resolution the schizophrenia cognitive impairment model, observation of injection compound l-slr can improve the spatial cognitive ability of mice, and compare the antipsychotic drug haloperidol for this role and classic. In extrapyramidal adverse reactions, the compounds of l-slr were compared with typical antipsychotic haloperidol and analogs of l-spd, determination the mice in the injection of haloperidol (0.8mg/kg), l-slr (10,30,60,80mg/kg) and l-spd (30,60,80mg/kg) after 45 and 90 minutes after the freezing time, different. The adverse reactions between the extrapyramidal system. In the research of the mechanism, determination of compound l-slr and inhibition of dopamine receptor affinity by competitive radioactive biological ligands. The experimental results in the spontaneous activity of mice in the experiment, 30 minutes after the injection of l-slr decreased activity before and after the time of recovery the result indicated that the compound l-slr has sedative effect, duration of this effect is 30 minutes, in the experimental design process to avoid the effect in this period of time. In the study of prepulse inhibition experiment, compared with the veh group, MK-801 (0.3mg/kg) can significantly reduce rat prepulse inhibition of startle reflex (f5,44=6.696 P0.0001), and l-slr (10mg/kg, 15mg/kg) injection and haloperidol (0.2mg/kg) can significantly improve the prepulse inhibition of injury caused by MK-801 (P0.01), indicating that the compound of l-slr and haloperidol on MK-801 The prepulse inhibition of injury are effective; in climbing experiment, compared with group veh, apomorphine (2mg/kg) subcutaneous injection can significantly increase mouse climbing time (f4,45=19.13, P0.0001), haloperidol (0.2mg/kg) and l-slr (30mg/kg) could significantly inhibit the climbing behavior induced by apomorphine, indicating that mice climbing behavior of compound l-slr and haloperidol on apomorphine induced effective social contact. In mice, compared with the veh group, the social contact behaviors of MK-801 can inhibit the growth of mice, the mice of the social contact time decreased significantly (f3,36=7.566, p=0.0084), and haloperidol typically cannot be gregarious the contact time of mice prolonged; correspondingly, the compound l-slr can significantly increase the social contact time of mice in the 30mg/kg (f5,54=6.285, p=0.0001), l-slr at the dose of 30mg/kg on sperm Schizophrenia negative symptoms effectively, and haloperidol is invalid. Cognitive deficits in schizophrenia Y maze, compared with group veh, the spatial cognitive ability of MK-801 could inhibit the mice (f3,36=10.60, P0.0001), haloperidol did not improve the spatial cognitive injury in mice; and the compound l-slr at the dose of 30mg/kg can significantly improve the spatial cognitive impairment in mice (f5,54=11.58, P0.0001, l-slr) that cognitive impairment at the dose of 30mg/kg can improve schizophrenia. In extrapyramidal adverse reactions, compared with the veh group, in 45 minutes after administration of haloperidol (0.8mg/kg) and l-spd (60mg/kg, 80mg/kg) to mice. The stupor time increased significantly (f8,64=13.63, p0.001); drug 90 minutes later, haloperidol and l-SPD (30mg/kg, 60 mg/kg, 80 mg/kg) wood stiff behavior cause still exist (F8,64=14.54, P0.001), and l- In SLR 30mg/kg 60, mg/kg does not produce side effects that stupor, l-SLR does not produce extrapyramidal in the effective dose of anti schizophrenia. The study on Mechanism of compound l-SLR in various parts of 18F-Fallypride into the mice quickly after distribution into mouse brains, but to give 18F-Fallypride quickly after the administration of l-SLR in mice in the distribution of 18F-Fallypride in various parts of the brain after 15 minutes of drug administration were significantly lower than group FP (P0.05), which shows that the compound l-SLR can D2 and D3 receptor in the brain and the combination, which can combine with the antagonism of 18F-Fallypride. Conclusion: the spontaneous activity test shows that compound l-SLR has sedative effects and calm in a relatively short time, so we in the efficacy of the compound in the study to avoid in this period of time. Before experiment and experiment of pulse suppression can be seen climbing The compound l-SLR can, the positive symptoms of schizophrenia. The conclusion from the social avoidance experiment, compound l-SLR on the negative symptoms of schizophrenia. From the Y maze test can be seen, the compound l-SLR can improve the cognitive impairment of schizophrenia. Therefore, in the treatment of cognitive impairment of schizophrenia, l-SLR better than haloperidol. In extrapyramidal adverse reactions, compound l-SLR does not cause mice stupor in the effective dose, indicating less compound l-SLR induced extrapyramidal adverse reactions, compared with haloperidol and l-SPD have significant advantages. In the study of receptor binding in the various brain regions through antagonism of 18F-Fallypride radioactivity decreased, indicating the compound l-SLR can and dopamine D2, D3 receptors.

【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R965

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