内吗啡肽类似物的设计合成、生物活性筛选及类药性化合物库的构建
发布时间：2019-06-28 19:42
【摘要】：阿片受体属于G蛋白偶联受体家族,包括μ、δ、κ三种经典的受体以及ORL1受体。其中μ受体介导了强效的镇痛作用,但一般与耐受、依赖性和药物滥用等副作用有关；6受体缓解疼痛的功效不强,但却可以减少潜在的上瘾性,也不易发生耐受；κ受体可以介导外周组织的镇痛,并与神经内分泌和免疫调节有关；ORL1受体则与机体的自稳、神经内分泌及自主神经系统调节有关。吗啡生物碱作为常用的镇痛药物,具有严重的耐受性、成瘾性和呼吸抑制等副作用。内吗啡肽-1及内吗啡肽-2对μ受体的结合能力强、选择性高,并且在镇痛的同时不产生一些吗啡样的副作用。但是许多缺点却限制了其在临床上的应用,包括副作用、酶代谢不稳定以及透过血脑屏障能力低等。许多研究证明μ受体和6受体可以形成二聚体,δ受体激动剂可以增强μ受体激动剂的镇痛活性和有效性,而δ受体拮抗剂可以防止或者减少μ受体激动剂所产生的耐受和身体依赖性。因而,同时作用于μ受体与δ受体的双重活性的多肽及拟肽能保持很好的镇痛作用且可以减少副作用。 本论文以内吗啡肽为基础,通过引入不同的α-羟基-β-氨基酸或者α-肼基氨基酸合成相应的β-拟肽化合物,旨在开发新的基于内吗啡肽结构的阿片配体,提高酶解稳定性,在保持强镇痛活性的同时能够减少吗啡样耐受性的产生。 首先,本文以α-羟基-β-拟肽3为先导化合物,辅助以计算机分子柔性对接,通过液相合成α-羟基-β-氨基酸和位置扫描法共设计合成了49个内吗啡肽类似物,并进行了一系列体外和体内生物筛选：阿片受体结合实验表明部分化合物对μ受体的亲和力与内吗啡肽相当,当同时引入Dmt1-Tic2后,化合物对μ受体和δ受体的作用均很强,可能成为潜在的具有μ/δ受体双重活性的拟肽；代谢稳定性实验表明α-羟基-β-拟肽化合物的半衰期均有不同程度的提高；而体外功能性反应实验表明化合物34对δ受体具有部分激动活性；在小鼠甩尾试验中,化合物34的镇痛活性与吗啡相当；在不同的小鼠模型中,化合物34被证实为混合阿片受体激动剂和δ受体拮抗剂；同时,化合物34在100nmol的高剂量下不影响小鼠的活动能力,在急性耐受性模型中没有产生像吗啡样的急性耐受作用。最后,我们将相关化合物与μ受体进行分子对接,发现对μ受体选择性较好、亲和力较高的化合物(化合物19和48)与(β-FNA)作用模式相似；化合物34作为儿受体激动/δ受体拮抗作用的配体,通过与δ阿片受体的非活性型进行柔性对接后,作用模式与Naltrindole以及μ受体激动剂/δ受体拮抗剂DIPP-NH2极为相似。 与α-羟基-β-氨基酸相似的是我们在内吗啡肽中引入了α-肼基氨基酸,在固相上直接合成了α-肼基拟肽化合物,并进行了体外生物筛选。部分化合物对μ受体的抑制率较高(50%),而且部分化合物对μ受体和δ受体的抑制率均较高(50%)。另外,该类化合物对不同阿片受体的亲和力数据表明,a-肼基氨基酸取代基的种类和在内吗啡肽中的位置十分重要。与内吗啡肽相比,大部分化合物对μ受体的亲和力均大幅度降低,但是化合物M-29和M-51对μ受体的亲和力与内吗啡肽相当。以设计合成具有双重活性的拟肽为基础,同时引入了Dmt1-Tic2后发现,大部分化合物保留了对δ受体的亲和力,对μ受体的亲和力均不同程度的下降。但化合物M-55对μ受体的亲和力与DIPP-NH2相当,对δ受体的亲和力相对DIPP-NH2提高了4倍。除此之外,代谢稳定性实验表明该类α-肼基拟肽化合物的稳定性比内吗啡肽提高了很多。同时,化合物M-55与δ阿片受体的非活性型进行柔性对接后,作用模式与Naltrindole以及μ受体激动剂/6受体拮抗剂DIPP-NH2极为相似。化合物M-55极有可能是一个μ受体激动剂/δ受体拮抗剂。该类化合物的生物活性筛选还在进行中。 综上所述,无论是α-羟基-β-拟肽还是α-肼基拟肽,在保持镇痛活性的同时,均能提高内吗啡肽的酶解稳定性,并且有可能提高其生物利用度,目前后续的生物筛选也正在进行中。糖基化与脂化是提高血脑屏障通透性和生物利用度的有效手段之一,而上述两类化合物的活性羟基(α-羟基-β-拟肽)和氨基(α-肼基拟肽)是潜在的糖基化或脂化位点,有可能使内吗啡肽类化合物更加适于临床使用。总而言之,多肽类镇痛药物的研究和开发仍然面临着很大很多的挑战,期待在不久的将来能够开发出能够在临床上使用的新型多肽类镇痛药物,有效的同时不产生耐受等副作用,为广大患者带来福音。 化合物的高通量筛选仍是新药发现的主要手段之一,本人在博士期间有幸参与了国家化合物的构建,包括8个类药性化合物库共计519个化合物,其针对不同靶点的活性评价正在进行中。
[Abstract]:Opioid receptors belong to the G-protein-coupled receptor family, including the three classical receptors and the ORL1 receptor. in which the mu receptor has a strong analgesic effect, but is generally associated with side effects such as tolerance, dependence and drug abuse; the efficacy of the receptor for relieving pain is not strong, but it can reduce the potential addiction and is not easy to tolerate; the opioid receptor can mediate the pain of the peripheral tissue, And the ORL1 receptor is related to the homeostasis of the body, the neuroendocrine and the regulation of autonomic nervous system. The morphine alkaloid is used as a common analgesic drug, and has the side effects of severe tolerance, addiction and respiratory depression. The combination ability of the endomorphin-1 and the endomorphin-2 to the. mu. receptor is strong, the selectivity is high, and the side effect of some morphine-like is not generated at the same time of the analgesia. However, many of the disadvantages limit their clinical use, including side effects, unstable enzyme metabolism, and low blood-brain barrier ability. Many studies have shown that the. mu. receptor and the 6 receptor can form a dimer, which can enhance the analgesic activity and effectiveness of the. mu. receptor agonist, while the opioid receptor antagonist can prevent or reduce the tolerance and physical dependence of the. mu. receptor agonist. Thus, the polypeptide and the peptidomimetic which simultaneously act on the dual activity of the. mu. receptor and the antigen receptor can maintain a good analgesic effect and can reduce the side effect. Based on the morphine peptide, the corresponding opioid-peptidomimetic compounds are synthesized by the introduction of different p-hydroxy-1-amino acids or the p-amino-base amino acids, aiming at the development of new opioid ligands based on the structure of the endomorphine peptide, and the stability of the enzymolysis is improved. The ability to reduce morphine-like tolerance while maintaining strong analgesic activity In this paper, a total of 49 endomorphins were synthesized by the co-design of the 1-hydroxy-1-pseudopeptide 3 as the lead compound and the aid of the flexible butt joint of the computer and the co-design of the 1-hydroxy-1-amino acid and the position scanning method by the liquid-phase synthesis. Analogues and a series of in vitro and in vivo biological screening: opioid receptor binding experiments show that the affinity of some of the compounds to the. mu. receptor is comparable to that of the endomorphine peptide, and the effect of the compound on the. mu. receptor and the opioid receptor after the introduction of the Dmt1-Tic2 at the same time The results of the in vitro functional response show that the compound 34 has a partial shock to the opioid receptor. kinetic activity; in a mouse flick test, the analgesic activity of compound 34 is comparable to morphine; in different mouse models, compound 34 is confirmed as a mixed opioid receptor agonist and a opioid receptor antagonist; at the same time, compound 34 does not affect the activity of the mouse at a high dose of 100 nmol Dynamic capacity, no acute tolerance to morphine-like in the acute tolerance model In that end, the compound (compound 19 and 48) with high selectivity and high affinity to the. mu. receptor was found to be similar to the (e-FNA) mode of action, and the compound 34, as the receptor agonist/ receptor antagonist, was found to be similar to the (e-FNA) mode of action. Ligand, by flexible docking with the non-active form of opioid receptors, the mode of action is associated with the Naltrindole and the. mu. receptor agonist/ antigen receptor antagonist DIPP-NH2 It is similar to that of the p-hydroxy-1-amino acid, which is the introduction of the p-amino-base amino acid in the endomorphin, which is directly synthesized on the solid phase, and the body is carried out. The inhibitory rate of some of the compounds on the receptor was higher (50%), and the inhibitory rate of some of the compounds on the receptor and the receptor was higher. (50%). In addition, the affinity data for these compounds for different opioid receptors indicate the type of a-tribasic amino acid substituent and the position of the internal morphine peptide It is very important to set the affinity of the compound M-29 and M-51 to the. mu. receptor in comparison with the endomorphin, but the affinity of the compounds M-29 and M-51 to the. The morphine peptide was comparable. On the basis of the design of a pseudo-peptide with a double activity, it was found that most of the compounds retained the affinity for the opioid receptor and the affinity for the. m However, the affinity of the compound M-55 to the. m. receptor is comparable to that of the DIPP-NH2, and the affinity of the compound M-55 with respect to the antigen receptor is relatively to the DIIPP-NH2. In addition to this, the metabolic stability experiments show that the stability of the class of opioid-based peptidomimetic compounds is higher than that of the endomorphin In addition, after the flexible docking of the compound M-55 with the non-active type of the opioid receptor, the mode of action is associated with the Naltrindole and the. mu. receptor agonist/6 receptor antagonist DIPP-NH. 2 Very similar. Compound M-55 is highly likely to be a. mu. receptor agonist/ antagonist Receptor Antagonists. Bioactivity screening of such compounds It is also in progress. In view of the above, it is possible to improve the enzymatic hydrolysis stability of the endomorphin at the same time as the analgesic activity is maintained, and it is possible to improve the bioavailability of the endomorphin at the same time as the analgesic activity is maintained, and the subsequent bioscreen Selection is also in progress. Glycosylation and lipogenesis are one of the most effective means to increase the permeability and bioavailability of the blood-brain barrier, while the active hydroxyl groups of the above-mentioned two classes of compounds (p-hydroxy-co-peptidomimetics) and amino (e-tribasic peptidomimetics) are potential sugars. the ethylated or lipidated site, it is possible to make the endomorphin compound more in general, that research and development of the polypeptide-like analgesic drug still face a great deal of challenge, and expect to develop novel polypeptide-like analgesic drugs which can be used clinically in the near future, The high-throughput screening of compounds is still one of the main means of new drug discovery, and I have the privilege of participating in the construction of the national compound during the doctor's period, including the total of 519 compounds in the eight drug-based compound libraries, which target different targets
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R914.5
本文编号：2507561
[Abstract]:Opioid receptors belong to the G-protein-coupled receptor family, including the three classical receptors and the ORL1 receptor. in which the mu receptor has a strong analgesic effect, but is generally associated with side effects such as tolerance, dependence and drug abuse; the efficacy of the receptor for relieving pain is not strong, but it can reduce the potential addiction and is not easy to tolerate; the opioid receptor can mediate the pain of the peripheral tissue, And the ORL1 receptor is related to the homeostasis of the body, the neuroendocrine and the regulation of autonomic nervous system. The morphine alkaloid is used as a common analgesic drug, and has the side effects of severe tolerance, addiction and respiratory depression. The combination ability of the endomorphin-1 and the endomorphin-2 to the. mu. receptor is strong, the selectivity is high, and the side effect of some morphine-like is not generated at the same time of the analgesia. However, many of the disadvantages limit their clinical use, including side effects, unstable enzyme metabolism, and low blood-brain barrier ability. Many studies have shown that the. mu. receptor and the 6 receptor can form a dimer, which can enhance the analgesic activity and effectiveness of the. mu. receptor agonist, while the opioid receptor antagonist can prevent or reduce the tolerance and physical dependence of the. mu. receptor agonist. Thus, the polypeptide and the peptidomimetic which simultaneously act on the dual activity of the. mu. receptor and the antigen receptor can maintain a good analgesic effect and can reduce the side effect. Based on the morphine peptide, the corresponding opioid-peptidomimetic compounds are synthesized by the introduction of different p-hydroxy-1-amino acids or the p-amino-base amino acids, aiming at the development of new opioid ligands based on the structure of the endomorphine peptide, and the stability of the enzymolysis is improved. The ability to reduce morphine-like tolerance while maintaining strong analgesic activity In this paper, a total of 49 endomorphins were synthesized by the co-design of the 1-hydroxy-1-pseudopeptide 3 as the lead compound and the aid of the flexible butt joint of the computer and the co-design of the 1-hydroxy-1-amino acid and the position scanning method by the liquid-phase synthesis. Analogues and a series of in vitro and in vivo biological screening: opioid receptor binding experiments show that the affinity of some of the compounds to the. mu. receptor is comparable to that of the endomorphine peptide, and the effect of the compound on the. mu. receptor and the opioid receptor after the introduction of the Dmt1-Tic2 at the same time The results of the in vitro functional response show that the compound 34 has a partial shock to the opioid receptor. kinetic activity; in a mouse flick test, the analgesic activity of compound 34 is comparable to morphine; in different mouse models, compound 34 is confirmed as a mixed opioid receptor agonist and a opioid receptor antagonist; at the same time, compound 34 does not affect the activity of the mouse at a high dose of 100 nmol Dynamic capacity, no acute tolerance to morphine-like in the acute tolerance model In that end, the compound (compound 19 and 48) with high selectivity and high affinity to the. mu. receptor was found to be similar to the (e-FNA) mode of action, and the compound 34, as the receptor agonist/ receptor antagonist, was found to be similar to the (e-FNA) mode of action. Ligand, by flexible docking with the non-active form of opioid receptors, the mode of action is associated with the Naltrindole and the. mu. receptor agonist/ antigen receptor antagonist DIPP-NH2 It is similar to that of the p-hydroxy-1-amino acid, which is the introduction of the p-amino-base amino acid in the endomorphin, which is directly synthesized on the solid phase, and the body is carried out. The inhibitory rate of some of the compounds on the receptor was higher (50%), and the inhibitory rate of some of the compounds on the receptor and the receptor was higher. (50%). In addition, the affinity data for these compounds for different opioid receptors indicate the type of a-tribasic amino acid substituent and the position of the internal morphine peptide It is very important to set the affinity of the compound M-29 and M-51 to the. mu. receptor in comparison with the endomorphin, but the affinity of the compounds M-29 and M-51 to the. The morphine peptide was comparable. On the basis of the design of a pseudo-peptide with a double activity, it was found that most of the compounds retained the affinity for the opioid receptor and the affinity for the. m However, the affinity of the compound M-55 to the. m. receptor is comparable to that of the DIPP-NH2, and the affinity of the compound M-55 with respect to the antigen receptor is relatively to the DIIPP-NH2. In addition to this, the metabolic stability experiments show that the stability of the class of opioid-based peptidomimetic compounds is higher than that of the endomorphin In addition, after the flexible docking of the compound M-55 with the non-active type of the opioid receptor, the mode of action is associated with the Naltrindole and the. mu. receptor agonist/6 receptor antagonist DIPP-NH. 2 Very similar. Compound M-55 is highly likely to be a. mu. receptor agonist/ antagonist Receptor Antagonists. Bioactivity screening of such compounds It is also in progress. In view of the above, it is possible to improve the enzymatic hydrolysis stability of the endomorphin at the same time as the analgesic activity is maintained, and it is possible to improve the bioavailability of the endomorphin at the same time as the analgesic activity is maintained, and the subsequent bioscreen Selection is also in progress. Glycosylation and lipogenesis are one of the most effective means to increase the permeability and bioavailability of the blood-brain barrier, while the active hydroxyl groups of the above-mentioned two classes of compounds (p-hydroxy-co-peptidomimetics) and amino (e-tribasic peptidomimetics) are potential sugars. the ethylated or lipidated site, it is possible to make the endomorphin compound more in general, that research and development of the polypeptide-like analgesic drug still face a great deal of challenge, and expect to develop novel polypeptide-like analgesic drugs which can be used clinically in the near future, The high-throughput screening of compounds is still one of the main means of new drug discovery, and I have the privilege of participating in the construction of the national compound during the doctor's period, including the total of 519 compounds in the eight drug-based compound libraries, which target different targets
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R914.5
【参考文献】
相关期刊论文 前1条
1 ;Endomorphin-1 and -2 inhibit human vascular sympathetic norepinephrine release:lack of interaction with μ_3 opiate receptor subtype[J];Acta Pharmacologica Sinica;1998年05期
，本文编号：2507561
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