新型阿片与神经肽FF多靶点多肽的化学筛选及药理学活性研究

发布时间：2018-02-27 10:44

本文关键词： 镇痛药慢性疼痛多靶点激动剂神经肽FF(NPFF) 阿片耐受　出处：《兰州大学》2017年博士论文　论文类型：学位论文

【摘要】：疼痛是一种不愉快的感觉或者情绪体验,生理条件下的疼痛对我们的身体是一种保护机制,能够提醒我们存在潜在的伤害或者疾病。然而,慢性疼痛却能够显著影响人们的生活质量,并且全球每年有数千万人遭受着各种各样的疼痛。疼痛的治疗,尤其是慢性疼痛的治疗是临床上面临的一个重大挑战。尽管阿片类镇痛药物在治疗各种急重性和慢性疼痛中起到了重要的作用,但是目前的阿片类镇痛药物具有一系列的副作用,因而极大地限制了它们在临床上尤其在慢性疼痛治疗中的应用。因此,镇痛效果好且毒副作用小的新型镇痛药物的研发需求已经越来越迫切。最近的研究发现,多靶点的阿片类镇痛药物研发可能是减少阿片副作用一个非常有效的策略。已有的研究表明神经肽FF(NPFF)对阿片的镇痛、镇痛耐受以及成瘾等多种活性具有重要的调节作用。我们近期的研究报道了一种基于阿片肽和NPFF关键药效团构建的嵌合肽BN-9,其能够产生高效、无耐受的镇痛作用。然而,多靶点分子对每个靶标活性的平衡对其药效以及可能的副作用具有重要的影响。为了阐述多靶点分子BN-9的构效关系,同时获得更高效且低副作用的新型阿片/NPFF多靶点分子,本论文设计合成了11个BN-9的类似物,并且对这些类似物的在体和离体的药理学活性进行了鉴定。在体外cAMP功能实验中,这些类似物均表现为阿片受体和NPFF受体的多靶点激动剂。在小鼠甩尾实验中,除类似物2和3以外,其余类似物侧脑室给药后都能产生与母肽BN-9相当或更强的无耐受镇痛作用。其中类似物11(DN-9)的镇痛作用是BN-9的33倍。侧脑室注射DN-9产生的镇痛作用能被μ和δ阿片受体的选择性拮抗剂β-FNA和nor-BNI完全拮抗,而NPFF受体拮抗剂能增强DN-9的镇痛作用。以上结果说明DN-9在脑室水平的镇痛作用主要由μ和κ阿片受体介导,同时NPFF受体在该镇痛作用中起到抗阿片的作用。另外,侧脑室注射DN-9在福尔马林痛模型和CFA诱导的慢性炎症痛模型中都具有很强的镇痛作用。值得一提的是,在甩尾实验以及CFA诱导的慢性炎症痛模型中,侧脑室连续6天给药,DN-9都不会产生镇痛耐受,同时也不会与吗啡产生交叉耐受现象。在脊髓水平,DN-9在多种临床前疼痛模型中都表现出非常强的镇痛作用。在小鼠甩尾实验中,DN-9的镇痛作用是BN-9的218倍,是吗啡的263倍,同时连续给药不会产生镇痛耐受。拮抗实验的结果与脑室水平相似,DN-9在脊髓水平的镇痛作用主要是由阿片受体介导,并且NPFF受体也参与了DN-9的脊髓镇痛。鞘内注射DN-9在手术后痛、醋酸扭体、福尔马林痛、角叉藻多糖诱导的炎症痛以及神经痛模型中都具有良好的镇痛作用。DN-9在这些痛模型中的镇痛的ED50值都在pmol级。本论文同时也探讨了DN-9在系统水平的镇痛作用。腹腔注射DN-9在小鼠甩尾实验、醋酸扭体、福尔马林痛以及手术后痛等疼痛模型中均能产生显著的镇痛作用。使用阿片受体和NPFF受体拮抗剂的实验结果表明,腹腔注射DN-9产生的镇痛作用主要通过阿片受体介导,而NPFF系统不参与DN-9在系统水平的镇痛作用。此外,DN-9在胃肠运动抑制、条件位置偏爱、心血管调节、体温调节以及运动功能和协调性等方面都表现出较低的副作用。总的来说,本研究中,我们首先设计合成并且药理学鉴定了一系列阿片/NPFF受体多靶点激动剂BN-9的类似物。我们进一步筛选出DN-9,其能够产生高效、无耐受的镇痛,并且消除或者减弱了一些阿片类镇痛药物常见的副作用。DN-9可以作为研发高效、低副作用的多靶点阿片类镇痛药物的一个非常好的前体化合物。本研究同时也证明了可以将阿片肽和NPFF的关键药效团嵌合在同一个分子中从而产生高效、无耐受的镇痛作用。
[Abstract]:Pain is an unpleasant sensation or emotion experience and physiological conditions of pain is a protective mechanism for our body, to remind us of the potential injury or illness. However, chronic pain can significantly affect the quality of people's lives, and the world every year tens of millions of people suffer from a variety of pain. The treatment of pain, especially in the treatment of chronic pain is a major clinical challenge facing. Although opioid analgesics in the treatment of various acute and chronic pain has played an important role, but with a series of side effects of the opioid analgesic drugs, which greatly limits their in clinic, especially in the treatment of chronic pain in the application. Therefore, a good analgesic effect and new analgesic drugs with little toxicity research becomes more and more urgent. A recent study found that multiple targets The opioid analgesic drug development may reduce opioid side effects of a very effective strategy. Studies have shown that neuropeptide FF (NPFF) on opioid analgesia, analgesic tolerance and addiction and other activities play an important role. Our recent studies reported a chimeric peptide opioid peptide and BN-9 NPFF key pharmacophore model based on it can produce efficient analgesia without tolerance. However, the balance of multi target molecules for each target activity has a significant influence on the efficacy and possible side effects. In order to explain the structure-activity relationship of target molecule BN-9, and obtain new opioid /NPFF more efficient low side effects of multiple target molecules, 11 BN-9 analogues were designed and synthesized in this paper, and these analogues in vitro and pharmacology activity were identified. The function of cAMP in vitro experiments, these similar Things were targets of opioid receptor and NPFF receptor agonist. In the tail flick test, in addition to analogs 2 and 3, the remaining analogue intracerebroventricular administration can produce analgesic effect without tolerance with the parent peptide BN-9 equivalent or stronger. Among the 11 analogues (DN-9) analgesia effect is 33 times that of BN-9. The analgesic effect of intracerebroventricular injection of DN-9 can be produced by selective antagonist Mu and delta opioid receptors -FNA and nor-BNI completely antagonistic agent beta, and NPFF receptor antagonists can enhance the analgesic effect of DN-9. The above results showed that the analgesic effect of DN-9 on ventricular level mainly by opioid receptor mediated by NPFF receptor in the analgesic effect plays anti opioid effect. In addition, it has strong analgesic effect of the chronic inflammatory pain model of intracerebroventricular injection of DN-9 in formalin pain model induced by CFA. It is worth mentioning that the induction in the tail flick test and CFA The chronic inflammatory pain model, lateral ventricle was administered for 6 consecutive days, DN-9 will not produce analgesia tolerance, but also will not cross with morphine tolerance. At the spinal cord level, DN-9 in a variety of preclinical models of pain showed analgesic effect is very strong. In the tail flick test, the analgesic effect of DN-9 is BN-9 218 times, 263 times of morphine analgesic tolerance, while continuing to produce drug not. The antagonistic experiment result is similar to ventricular level, the analgesic effect of DN-9 in spinal cord is mainly mediated by the opioid receptors, and NPFF receptors in the DN-9 spinal analgesia. Intrathecal injection of DN-9 in postoperative pain formalin, acetic acid, and has a good analgesic effect of.DN-9 in the analgesic pain model in carrageenan polysaccharide induced inflammatory pain and neuropathic pain models in the ED50 value at the pmol level. The paper also discusses the DN-9 in the Department The level of analgesia in the tail flick test. Intraperitoneal injection of DN-9, acetic acid, formalin and pain after the operation pain model can generate significant analgesic effects. The experimental results using the opioid receptor and NPFF receptor antagonists show that the analgesic effect of intraperitoneal injection of DN-9 is mainly mediated by opioid receptors, while the NPFF system is not involved in the analgesic effect of DN-9 in system level. In addition, DN-9 in the inhibition of gastrointestinal motility, CPP, cardiovascular regulation, thermoregulation and motion function and coordination in terms of lower side effects. In general, in this study, we first design and synthesis and identification of pharmacology a series of analogues of /NPFF opioid receptor agonist BN-9 multi targets. We screened DN-9, which can produce high tolerance, no pain, and eliminate or weaken some opioid town .DN-9 pain drug common side effects can be used to develop an efficient, a very good precursor compound target low side effects of opioid drugs. This study also demonstrated that opioids and NPFF can be the key to produce high efficiency chimeric pharmacophore in the same molecule, analgesia the role of tolerance.

【学位授予单位】：兰州大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R96

【相似文献】