本文選題：p53-MDM2　切入點(diǎn)：AMG232　出處：《寧夏醫(yī)科大學(xué)》2017年碩士論文

【摘要】：抑癌基因p53是一種在腫瘤發(fā)生中起著重要作用的調(diào)節(jié)因子,被廣泛認(rèn)為是“基因組的守護(hù)者”,參與受損細(xì)胞的修復(fù)和凋亡。MDM2蛋白是p53最重要的負(fù)反饋調(diào)節(jié)因子。MDM2過(guò)量表達(dá)會(huì)導(dǎo)致細(xì)胞增殖失控,繼而導(dǎo)致腫瘤生長(zhǎng)。AMG系列化合物是安進(jìn)公司報(bào)道的哌啶酮類小分子抑制劑,可以通過(guò)與MDM2特定位點(diǎn)結(jié)合,阻斷p53-MDM2相互作用,對(duì)腫瘤細(xì)胞增殖具有很高的抑制活性。AMG232是其中活性最好的小分子抑制劑之一,分子和細(xì)胞水平抑制率都達(dá)到了低nM級(jí),目前已進(jìn)入臨床II期試驗(yàn),有望開(kāi)發(fā)成新的抗腫瘤藥物。但是其羧基側(cè)鏈在體內(nèi)容易通過(guò)葡萄糖醛酸結(jié)合途徑代謝失活,導(dǎo)致其半衰期較短,成藥后可能會(huì)給患者造成頻繁給藥的負(fù)擔(dān)。本課題組擬通過(guò)將AMG232羧基側(cè)鏈改造為酯類,降低其在體內(nèi)與葡萄糖醛酸結(jié)合而代謝失活的能力,延長(zhǎng)其半衰期,改善藥代動(dòng)力學(xué)性質(zhì)。設(shè)計(jì)合成得到15個(gè)全新結(jié)構(gòu)的AMG232酯類衍生物。其中化合物7的蛋白結(jié)合活性與對(duì)HCT116癌細(xì)胞系的抑制活性最好,與陽(yáng)性藥相當(dāng)。另外,我們對(duì)AMG232關(guān)鍵中間體的合成路線進(jìn)行了初步探索,設(shè)計(jì)打通了一條收率高、反應(yīng)條件溫和的AMG232關(guān)鍵中間體的合成路線。研究發(fā)現(xiàn),NF-κB基因突變激活與決定許多腫瘤進(jìn)展的慢性炎癥息息相關(guān),而且還會(huì)誘導(dǎo)腫瘤細(xì)胞對(duì)多種抗腫瘤藥物產(chǎn)生耐藥性。我們課題組前期通過(guò)虛擬篩選得到一類同時(shí)激活p53信號(hào)通路和抑制NF-κB信號(hào)通路全新結(jié)構(gòu)類型的高活性吡咯酮類化合物,并基于吡咯酮骨架先導(dǎo)物進(jìn)行了合理藥物設(shè)計(jì)發(fā)現(xiàn)了活性更強(qiáng)的吡咯酮并吡唑化合物。機(jī)制研究表明,R-異構(gòu)體能激活p53,并呈劑量依賴關(guān)系,同時(shí)抑制MDM2的表達(dá)。而S-異構(gòu)體則對(duì)p53通路沒(méi)有明顯的作用。對(duì)NF-κB信號(hào)通路,這對(duì)異構(gòu)體的作用則與p53通路完全相反,S-異構(gòu)體可以激活磷酸化的IKKα、β、γ而抑制NF-κB的激活,而R-異構(gòu)體則對(duì)NF-κB通路沒(méi)有影響。為進(jìn)一步研究光學(xué)異構(gòu)體對(duì)活性的影響,我們對(duì)吡咯酮類化合物進(jìn)行不對(duì)稱合成研究,獲得了4對(duì)高ee值的R、S-光學(xué)異構(gòu)體。在此基礎(chǔ)上,通過(guò)體外抗腫瘤活性研究,獲得了兩種光學(xué)異構(gòu)體的最佳配比,為后續(xù)的p53-MDM2與NF-κB信號(hào)通路交互協(xié)同作用研究打下堅(jiān)實(shí)的基礎(chǔ)。
[Abstract]:The tumor suppressor gene p53 is a regulatory factor that plays an important role in tumorigenesis. It is widely regarded as a "guardian of the genome", which is involved in the repair and apoptosis of damaged cells. MDM2 protein is the most important negative feedback regulator of p53. MDM2 overexpression can lead to uncontrolled cell proliferation. The resulting tumor growth. AMG series of compounds are small molecular inhibitors of piperidone reported by Amgen, which can block p53-MDM2 interactions by binding to specific sites of MDM2. AMG232 is one of the best small molecular inhibitors of tumor cell proliferation. The inhibition rate of both molecular and cell level has reached low NM level, and has entered the phase II clinical trial. It is expected to be a new antitumor drug, but its carboxyl side chain is easily metabolized by glucuronic acid binding pathway in vivo, resulting in a shorter half-life. The study group intends to transform the carboxyl side chain of AMG232 into esters so as to reduce its ability of metabolism inactivation by binding with glucuronic acid in vivo and prolong its half-life. To improve the pharmacokinetic properties, 15 novel AMG232 ester derivatives were designed and synthesized. The protein-binding activity of compound 7 was the best and the inhibitory activity was similar to that of the positive drug. The synthetic route of the key intermediate of AMG232 was preliminarily explored and a high yield was designed. Synthesis of key Intermediates of AMG232 with mild reaction conditions. It has been found that the activation of NF- 魏 B gene mutation is closely related to the chronic inflammation that determines the progression of many tumors. Moreover, it can induce tumor cells to develop drug resistance to various antitumor drugs. Our team obtained a class of highly active pyrrolidone compounds that simultaneously activate p53 signaling pathway and inhibit new structural types of NF- 魏 B signaling pathway through virtual screening. The more active pyrrolidone benzopyrazole compounds were found based on the pyrrolidone skeleton lead. The mechanism showed that the pyrrolidone isomer could activate p53 in a dose-dependent manner. At the same time, the expression of MDM2 was inhibited. The S- isomer had no obvious effect on p53 pathway, but on NF- 魏 B signaling pathway, the effect of the isomer was opposite to that of p53 pathway. S- isomer could activate phosphorylated IKK 偽, 尾, 緯 and inhibit the activation of NF- 魏 B. However, R- isomer has no effect on NF- 魏 B pathway. In order to further study the effect of optical isomers on activity, we have studied the asymmetric synthesis of pyrrolidone compounds and obtained 4 pairs of high ee RGS-optical isomers. The optimal ratio of two optical isomers was obtained through in vitro antitumor activity study, which laid a solid foundation for the further study of the interaction between p53-MDM2 and NF- 魏 B signaling pathway.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R914

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 覃凌云;陳蓉;蘇正定;;Mdm2/MdmX抑制劑[J];中國(guó)生物工程雜志;2015年09期

2 高梅;楊桂文;;ARF——重要的腫瘤抑制蛋白[J];科技信息(學(xué)術(shù)研究);2008年36期

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本文編號(hào)：1655233