【摘要】：阿片受體屬于G蛋白偶聯(lián)受體家族,包括μ、δ、κ三種經(jīng)典的受體以及ORL1受體。其中μ受體介導(dǎo)了強(qiáng)效的鎮(zhèn)痛作用,但一般與耐受、依賴性和藥物濫用等副作用有關(guān)；6受體緩解疼痛的功效不強(qiáng),但卻可以減少潛在的上癮性,也不易發(fā)生耐受；κ受體可以介導(dǎo)外周組織的鎮(zhèn)痛,并與神經(jīng)內(nèi)分泌和免疫調(diào)節(jié)有關(guān)；ORL1受體則與機(jī)體的自穩(wěn)、神經(jīng)內(nèi)分泌及自主神經(jīng)系統(tǒng)調(diào)節(jié)有關(guān)。嗎啡生物堿作為常用的鎮(zhèn)痛藥物,具有嚴(yán)重的耐受性、成癮性和呼吸抑制等副作用。內(nèi)嗎啡肽-1及內(nèi)嗎啡肽-2對(duì)μ受體的結(jié)合能力強(qiáng)、選擇性高,并且在鎮(zhèn)痛的同時(shí)不產(chǎn)生一些嗎啡樣的副作用。但是許多缺點(diǎn)卻限制了其在臨床上的應(yīng)用,包括副作用、酶代謝不穩(wěn)定以及透過(guò)血腦屏障能力低等。許多研究證明μ受體和6受體可以形成二聚體,δ受體激動(dòng)劑可以增強(qiáng)μ受體激動(dòng)劑的鎮(zhèn)痛活性和有效性,而δ受體拮抗劑可以防止或者減少μ受體激動(dòng)劑所產(chǎn)生的耐受和身體依賴性。因而,同時(shí)作用于μ受體與δ受體的雙重活性的多肽及擬肽能保持很好的鎮(zhèn)痛作用且可以減少副作用。 本論文以內(nèi)嗎啡肽為基礎(chǔ),通過(guò)引入不同的α-羥基-β-氨基酸或者α-肼基氨基酸合成相應(yīng)的β-擬肽化合物,旨在開(kāi)發(fā)新的基于內(nèi)嗎啡肽結(jié)構(gòu)的阿片配體,提高酶解穩(wěn)定性,在保持強(qiáng)鎮(zhèn)痛活性的同時(shí)能夠減少嗎啡樣耐受性的產(chǎn)生。 首先,本文以α-羥基-β-擬肽3為先導(dǎo)化合物,輔助以計(jì)算機(jī)分子柔性對(duì)接,通過(guò)液相合成α-羥基-β-氨基酸和位置掃描法共設(shè)計(jì)合成了49個(gè)內(nèi)嗎啡肽類似物,并進(jìn)行了一系列體外和體內(nèi)生物篩選：阿片受體結(jié)合實(shí)驗(yàn)表明部分化合物對(duì)μ受體的親和力與內(nèi)嗎啡肽相當(dāng),當(dāng)同時(shí)引入Dmt1-Tic2后,化合物對(duì)μ受體和δ受體的作用均很強(qiáng),可能成為潛在的具有μ/δ受體雙重活性的擬肽；代謝穩(wěn)定性實(shí)驗(yàn)表明α-羥基-β-擬肽化合物的半衰期均有不同程度的提高；而體外功能性反應(yīng)實(shí)驗(yàn)表明化合物34對(duì)δ受體具有部分激動(dòng)活性；在小鼠甩尾試驗(yàn)中,化合物34的鎮(zhèn)痛活性與嗎啡相當(dāng)；在不同的小鼠模型中,化合物34被證實(shí)為混合阿片受體激動(dòng)劑和δ受體拮抗劑；同時(shí),化合物34在100nmol的高劑量下不影響小鼠的活動(dòng)能力,在急性耐受性模型中沒(méi)有產(chǎn)生像嗎啡樣的急性耐受作用。最后,我們將相關(guān)化合物與μ受體進(jìn)行分子對(duì)接,發(fā)現(xiàn)對(duì)μ受體選擇性較好、親和力較高的化合物(化合物19和48)與(β-FNA)作用模式相似；化合物34作為兒受體激動(dòng)/δ受體拮抗作用的配體,通過(guò)與δ阿片受體的非活性型進(jìn)行柔性對(duì)接后,作用模式與Naltrindole以及μ受體激動(dòng)劑/δ受體拮抗劑DIPP-NH2極為相似。 與α-羥基-β-氨基酸相似的是我們?cè)趦?nèi)嗎啡肽中引入了α-肼基氨基酸,在固相上直接合成了α-肼基擬肽化合物,并進(jìn)行了體外生物篩選。部分化合物對(duì)μ受體的抑制率較高(50%),而且部分化合物對(duì)μ受體和δ受體的抑制率均較高(50%)。另外,該類化合物對(duì)不同阿片受體的親和力數(shù)據(jù)表明,a-肼基氨基酸取代基的種類和在內(nèi)嗎啡肽中的位置十分重要。與內(nèi)嗎啡肽相比,大部分化合物對(duì)μ受體的親和力均大幅度降低,但是化合物M-29和M-51對(duì)μ受體的親和力與內(nèi)嗎啡肽相當(dāng)。以設(shè)計(jì)合成具有雙重活性的擬肽為基礎(chǔ),同時(shí)引入了Dmt1-Tic2后發(fā)現(xiàn),大部分化合物保留了對(duì)δ受體的親和力,對(duì)μ受體的親和力均不同程度的下降。但化合物M-55對(duì)μ受體的親和力與DIPP-NH2相當(dāng),對(duì)δ受體的親和力相對(duì)DIPP-NH2提高了4倍。除此之外,代謝穩(wěn)定性實(shí)驗(yàn)表明該類α-肼基擬肽化合物的穩(wěn)定性比內(nèi)嗎啡肽提高了很多。同時(shí),化合物M-55與δ阿片受體的非活性型進(jìn)行柔性對(duì)接后,作用模式與Naltrindole以及μ受體激動(dòng)劑/6受體拮抗劑DIPP-NH2極為相似�；衔颩-55極有可能是一個(gè)μ受體激動(dòng)劑/δ受體拮抗劑。該類化合物的生物活性篩選還在進(jìn)行中。 綜上所述,無(wú)論是α-羥基-β-擬肽還是α-肼基擬肽,在保持鎮(zhèn)痛活性的同時(shí),均能提高內(nèi)嗎啡肽的酶解穩(wěn)定性,并且有可能提高其生物利用度,目前后續(xù)的生物篩選也正在進(jìn)行中。糖基化與脂化是提高血腦屏障通透性和生物利用度的有效手段之一,而上述兩類化合物的活性羥基(α-羥基-β-擬肽)和氨基(α-肼基擬肽)是潛在的糖基化或脂化位點(diǎn),有可能使內(nèi)嗎啡肽類化合物更加適于臨床使用�？偠灾�,多肽類鎮(zhèn)痛藥物的研究和開(kāi)發(fā)仍然面臨著很大很多的挑戰(zhàn),期待在不久的將來(lái)能夠開(kāi)發(fā)出能夠在臨床上使用的新型多肽類鎮(zhèn)痛藥物,有效的同時(shí)不產(chǎn)生耐受等副作用,為廣大患者帶來(lái)福音。 化合物的高通量篩選仍是新藥發(fā)現(xiàn)的主要手段之一,本人在博士期間有幸參與了國(guó)家化合物的構(gòu)建,包括8個(gè)類藥性化合物庫(kù)共計(jì)519個(gè)化合物,其針對(duì)不同靶點(diǎn)的活性評(píng)價(jià)正在進(jìn)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
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R914.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

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

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本文編號(hào)：2507561