本文關(guān)鍵詞： 海兔毒素10 合成 多肽 類似物 前藥 抗腫瘤活性　出處：《華東師范大學(xué)》2017年博士論文　論文類型：學(xué)位論文

【摘要】：海兔毒素10是從印度洋的海兔Dolabbella auriclaria體內(nèi)分離得到的一種海洋天然產(chǎn)物,它是一種高細(xì)胞毒性的多肽類化合物,對(duì)于多種腫瘤細(xì)胞都顯示出極強(qiáng)的抑制能力。雖然海兔毒素10作為抗腫瘤藥在臨床前研究顯示出良好的效果,但由于其在臨床研究中顯示出太大的毒性,故未能通過(guò)臨床試驗(yàn)。隨后,一系列被稱為"auristatins"的海兔毒素10衍生物被開(kāi)發(fā)出來(lái),在臨床試驗(yàn)研究中被大量應(yīng)用。而由其中一個(gè)化合物MMAE發(fā)展起來(lái)的抗體藥物偶聯(lián)物(ADC)"Adcetris"已經(jīng)作為治療霍奇金淋巴瘤的藥物于2011年被FDA批準(zhǔn)上市。此外,目前進(jìn)入臨床研究的ADC藥物有將近一半都是以海兔毒素10的衍生物作為藥效分子的,極有可能以此開(kāi)發(fā)得到新的抗腫瘤藥。另一方面,海兔毒素10有著復(fù)雜的多手性中心結(jié)構(gòu),其合成具有一定挑戰(zhàn),所以無(wú)論從化學(xué)合成的角度還是藥物化學(xué)的角度來(lái)看,對(duì)海兔毒素10及其類似物的合成做進(jìn)一步的研究都有著極為重要的意義。本論文的前言綜述了一些多肽以及環(huán)肽類化合物的合成方法研究進(jìn)展。主要介紹了具有抗腫瘤活性的天然海洋肽類化合物,重點(diǎn)介紹海兔毒素10的發(fā)現(xiàn)、合成以及其衍生物的發(fā)展。本論文的研究?jī)?nèi)容主要包括以下四個(gè)部分:第一部分,首先分別對(duì)海兔毒素10的三個(gè)關(guān)鍵中間體片段Dil,Dap以及Doe的合成進(jìn)行了深入的研究。通過(guò)對(duì)路線優(yōu)化和反應(yīng)條件篩選,基本上能夠高效大量地合成得到這三個(gè)關(guān)鍵中間體以及Dil和Dap片段的異構(gòu)體。隨后通過(guò)液相合成法完成了海兔毒素10以及其中三個(gè)手性異構(gòu)體的合成。第二部分,在目前已有文獻(xiàn)報(bào)道對(duì)海兔毒素10的構(gòu)效關(guān)系研究的基礎(chǔ)上,以海兔毒素10的衍生物MMAF為模板,對(duì)其進(jìn)行N-端結(jié)構(gòu)修飾。將N-端氨基酸的氨基修飾為伯胺,同時(shí)在氨基酸的αα-位引入不同的取代基,完成了 13個(gè)新的海兔毒素10類似物的合成。隨后又研究了構(gòu)象的改變對(duì)于海兔毒素10活性的影響,將合成得到的這一系列線性肽經(jīng)酰胺縮合環(huán)化,合成得到12個(gè)新的海兔毒素10環(huán)肽類似物。第三部分,針對(duì)海兔毒素10毒性太大這一特點(diǎn),將海兔毒素10設(shè)計(jì)為前藥的形式。主要是通過(guò)將其改造為環(huán)肽酯或者在分子結(jié)構(gòu)里引入酶可裂解型二肽接頭來(lái)實(shí)現(xiàn)。第四部分,對(duì)合成得到的海兔毒素10類似物進(jìn)行體外的抗腫瘤活性評(píng)估,通過(guò)測(cè)定這些化合物對(duì)HCT-116人結(jié)腸癌的的半數(shù)抑制濃度(IC50)來(lái)完成。測(cè)試結(jié)果顯示,其中10個(gè)N-端修飾的類似物都表現(xiàn)出比對(duì)照化合物MMAF更好的體外抗HCT-116細(xì)胞活性,活性最好的為化合物37f,IC50達(dá)到0.07μM。而對(duì)于環(huán)肽化合物,因?yàn)闃?gòu)象的改變,活性比相對(duì)應(yīng)的線性肽普遍降低,但是環(huán)肽化合物40k和401活性卻得到提高,IC50均達(dá)到0.03 μM,表現(xiàn)出了較強(qiáng)的抗腫瘤活性。
[Abstract]:Marine rabbit toxin 10 is a natural marine product isolated from Dolabbella auriclaria of the Indian Ocean. It is a highly cytotoxic polypeptide compound. Although the antitumor drug has shown a good effect in preclinical studies, it has shown too much toxicity in clinical research because of its strong inhibitory effect on many kinds of tumor cells. A series of Hare toxin 10 derivatives called "auristatins" were subsequently developed. It has been widely used in clinical trials. The antibody-drug coupling developed from one of the compounds, MMAE, is ADCA). "Adcetris" was approved by FDA on 2011 as a treatment for Hodgkin's lymphoma. At present, nearly half of the ADC drugs that enter the clinical research are using the derivatives of Hippotoxin 10 as effector molecules, which is very likely to be used to develop new anti-tumor drugs. On the other hand, it is possible to develop new antitumor drugs. Rabbit toxin 10 has a complex multi-chiral central structure, and its synthesis has some challenges, so it is not only from the perspective of chemical synthesis, but also from the point of view of pharmacochemistry. It is of great significance to further study the synthesis of sea rabbit toxin 10 and its analogues. The foreword of this paper summarizes the progress in the synthesis of some peptides and cyclic peptide compounds. Natural marine peptide compounds with antitumor activity. The discovery, synthesis and the development of its derivatives are introduced in detail. The research contents of this thesis mainly include the following four parts: the first part. Firstly, the synthesis of three key intermediate fragments of rabbit toxin 10, Dilo Dap and Doe, were studied, and the route optimization and reaction conditions were selected. The three key intermediates and the isomers of Dil and Dap fragments can be synthesized in large quantities by high efficiency. Subsequently, the synthesis of Hare toxin 10 and three of the chiral isomers were completed by liquid phase synthesis. Part two. On the basis of the research on the structure-activity relationship of Hippotoxin 10 in the literature, the derivative MMAF of Hippotoxin 10 was used as template. The amino groups of N-terminal amino acids were modified to primary amines. At the same time, different substituents were introduced into the 偽 -sites of amino acids. After the synthesis of 13 new mimics of toxin 10, the effects of conformational changes on the activity of toxin 10 were studied, and the synthesized linear peptides were cyclized by amide condensation. A total of 12 new cyclic peptide analogues were synthesized. The third part is aimed at the characteristic that the toxicity of Hippotoxin 10 is too high. It was designed as prodrug form, mainly by transforming it into cyclic peptide ester or introducing enzyme cleavable dipeptide junction into molecular structure to realize. 4th part. The antitumor activity of synthetic rabbit toxin 10 analogue was evaluated in vitro. This was done by measuring the half inhibitory concentration (IC50) of these compounds on HCT-116 human colon cancer. Ten of the N-terminal modified analogs showed better anti-#en1# cell activity than control compound MMAF in vitro, and the best activity was compound 37f. IC50 reached 0.07 渭 M. for cyclic peptide compounds, the activity of cyclic peptide compounds was generally lower than that of corresponding linear peptides due to conformation change, but the activities of cyclic peptide compounds 40k and 401were increased. IC50 reached 0.03 渭 M, showing strong anti-tumor activity.
【學(xué)位授予單位】：華東師范大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：O629

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相關(guān)期刊論文 前1條

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本文編號(hào)：1442581