發(fā)布時(shí)間：2018-03-26 01:21

  本文選題：sirtuin　切入點(diǎn)：SIRT5　出處：《江蘇大學(xué)》2017年碩士論文

【摘要】：蛋白翻譯后修飾(PTMs)越來(lái)越多的表明在調(diào)節(jié)蛋白功能以及在許多生理和病理過(guò)程中起了很大作用。賴氨酸乙�；潜谎芯孔疃嗟男揎椇驼{(diào)節(jié)之一,參與眾多至關(guān)重要的細(xì)胞內(nèi)過(guò)程包括蛋白降解,轉(zhuǎn)錄激活以及新陳代謝。沉默信息調(diào)節(jié)因子2(Sir2)或者說(shuō)sirtuin是一類(lèi)高度保守的依賴β-煙酰胺腺嘌呤二核苷酸(β-NAD+)的酶,它調(diào)節(jié)了許多代謝通路。自從2000年以來(lái),哺乳動(dòng)物中已經(jīng)發(fā)現(xiàn)了七個(gè)sirtuin家族成員,即SIRT1-7。Sirtuin被視為是當(dāng)代癌癥、代謝類(lèi)疾病以及神經(jīng)退行性疾病的治療靶點(diǎn)。因此,其抑制劑在過(guò)去這些年成為了研究重點(diǎn)。其中四個(gè)成員(SIRT4-7)沒(méi)有檢測(cè)到或者只檢測(cè)到很微弱的去乙�；钚�,傾向于去除長(zhǎng)鏈脂肪�；�、丁二�；�。在這份論文工作中,我們選用存在于線粒體和細(xì)胞質(zhì)中的SIRT5作為研究目標(biāo)。SIRT5是體內(nèi)高效的賴氨酸去丙二酰、去丁二酰、去戊二酰的去�；�。對(duì)于這種去除負(fù)電羧基的偏好可以解釋為由于105位的精氨酸殘基以及102位的酪氨酸殘基組成的人類(lèi)SIRT5去�；诖�。針對(duì)目前SIRT5抑制劑研究不夠充分的情況,更多高效且有選擇性的抑制劑需要被發(fā)現(xiàn)。首先,我們選用高效的催化機(jī)制導(dǎo)向的抑制彈頭---Nε-羧乙基-硫代氨甲酰-賴氨酸SIRT5抑制彈頭,將其運(yùn)用到我們?cè)O(shè)計(jì)的環(huán)肽骨架中去。設(shè)計(jì)合成了一系列帶有該彈頭的環(huán)肽化合物,用于抑制活性測(cè)試和構(gòu)效關(guān)系研究(SAR)。根據(jù)這些化合物SIRT5活性抑制測(cè)試的結(jié)果,這個(gè)環(huán)肽的抑制策略被證明是一個(gè)有效增強(qiáng)sirtuin抑制劑活性的方法�；衔�3發(fā)現(xiàn)是一個(gè)強(qiáng)效的SIRT5抑制劑(IC50=2.2±0.89μM),是相應(yīng)的直鏈肽化合物7的SIRT5抑制強(qiáng)度的42.3倍左右。此外,在蛋白酶水解測(cè)試中發(fā)現(xiàn),化合物3相較于直鏈肽化合物7其蛋白酶水解的耐受性很強(qiáng)。這也證明了環(huán)肽骨架的優(yōu)越性。再有,我們不滿足于現(xiàn)有的SIRT5抑制彈頭,嘗試性地設(shè)計(jì)了新型彈頭。將之前彈頭末端丙酸替換成了苯甲酸,向其中引入芳香環(huán),期待能更緊密占據(jù)SIRT5�；诖�。我們也將彈頭中硫脲結(jié)構(gòu)替換成硒脲,化合物9(IC50=1.7±0.1μM)相較于之前使用相同肽骨架的硫脲化合物(IC50=5±1.9μM)的SIRT5抑制活性提高了2.9倍左右。預(yù)期這一變化會(huì)成為整個(gè)sirtuin抑制劑研究的有力武器。
[Abstract]:Protein post-translational modification (PTMs) has been increasingly shown to play a significant role in regulating protein function and in many physiological and pathological processes. Lysine acetylation is one of the most widely studied modifications and modulations. Involved in a number of critical intracellular processes including protein degradation, transcriptional activation, and metabolism. Silencing signaling factor 2 / Sir2) or sirtuin is a highly conserved enzyme that relies on 尾 -nicotinamide adenine dinucleotides (尾 -NAD). It regulates many metabolic pathways. Since 2000, seven members of the sirtuin family, known as SIRT1-7.Sirtuin, have been identified as targets for modern cancer, metabolic diseases and neurodegenerative diseases. Its inhibitors have been the focus of research in the past few years. Four of its members, SIRT4-7, have not detected or only detected very weak deacetylation activities, and tend to remove long-chain fatty acyl, succinyl, and so on. We selected SIRT5 in mitochondria and cytoplasm as our research objective. SIRT5 is highly efficient lysine demalonyl and succinyl in vivo. De-glutaryl deacylase. The preference for the removal of negatively charged carboxyl groups can be explained by a human SIRT5 deacylation pocket consisting of 105th arginine residues and 102nd tyrosine residues. Insufficient circumstances, More efficient and selective inhibitors need to be found. First, we use a highly efficient catalytic mechanism directed inhibitory warhead-N 蔚 -carboxyethyl-thiocarbamate-lysine SIRT5 inhibitory warhead. A series of cyclic peptide compounds with this warhead were designed and synthesized for inhibition activity testing and structure-activity relationship study. Based on the results of SIRT5 activity inhibition tests of these compounds, The inhibition strategy of this cyclic peptide has been proved to be an effective method to enhance the activity of sirtuin inhibitor. Compound 3 was found to be a potent SIRT5 inhibitor, IC502.2 鹵0.89 渭 M-1, which is about 42.3 times of the SIRT5 inhibitory intensity of the corresponding straight chain peptide compound 7. In the proteolytic test, compound 3 was found to be more tolerant of proteolysis than that of straight chain peptide compound 7. This also proves the superiority of cyclic peptide skeleton. Moreover, we are not satisfied with the existing SIRT5 inhibitory warhead. A new warhead was tentatively designed. Propionic acid was replaced with benzoic acid at the end of the warhead, and aromatic rings were introduced into it, hoping to occupy more closely the SIRT5 acylation pocket. We also replaced the thiourea structure in the warhead with selenourea. The SIRT5 inhibitory activity of compound 9(IC50=1.7 鹵0.1 渭 M was about 2.9-fold higher than that of thiourea compound IC5050 鹵1.9 渭 M using the same peptide skeleton. It is expected that this change will become a powerful weapon in the study of sirtuin inhibitors.
【學(xué)位授予單位】：江蘇大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R91

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 Lingling Yang;Xiaobo Ma;Yanying He;Chen Yuan;Quanlong Chen;Guobo Li;Xianggui Chen;;Sirtuin 5:a review of structure,known inhibitors and clues for developing new inhibitors[J];Science China(Life Sciences);2017年03期

2 YANG Xin;LIU BoYa;ZHU WeiGuo;LUO JianYuan;;SIRT5, functions in cellular metabolism with a multiple enzymatic activities[J];Science China(Life Sciences);2015年09期

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本文編號(hào)：1665706