本文關(guān)鍵詞： sirtuin SIRT6 SIRT5 去�；� 抑制劑　出處：《江蘇大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：Sirtuin是一類依賴于β-NAD+的去除蛋白�；嚢彼醾�(cè)鏈�；娜ヵ；�,與其他去�；覆煌�,sirtuin催化的酰化賴氨酸側(cè)鏈去�；皇且粋�(gè)簡(jiǎn)單的酰胺水解反應(yīng),而是從煙酰胺腺嘌呤二核苷酸(β-NAD+或NAD+)上裂解一分子煙酰胺以生成三個(gè)產(chǎn)物(即去酰化的蛋白、煙酰胺和2’-O-AADP-核糖)的一個(gè)過(guò)程�，F(xiàn)在有越來(lái)越多的研究證明sirtuin催化的去酰化反應(yīng)能調(diào)節(jié)體內(nèi)多種重要的生物過(guò)程,同時(shí),該催化反應(yīng)也被認(rèn)為是多種人類疾病治療的潛在靶點(diǎn)。SIRT6是哺乳類動(dòng)物及人類中發(fā)現(xiàn)的7個(gè)sirtuin(SIRT1-7)之一,主要存在于細(xì)胞核中,除了催化去乙酰化反應(yīng),SIRT6具有更強(qiáng)的去脂肪酰(比如十四酰)的活性。SIRT6可以催化脫去組蛋白H3K9和H3K56上的乙�；鶑亩{(diào)控基因表達(dá)并且維持染色質(zhì)端粒。此外,SIRT6催化的去酰化反應(yīng)還與很多疾病息息相關(guān),如心血管疾病,癌癥,神經(jīng)退行性疾病和糖尿病等。SIRT5是一個(gè)主要存在于線粒體的酶,相比于催化去乙�；钚�,SIRT5具有更強(qiáng)的去丙二酰、去丁二酰和去戊二酰的活性。有研究發(fā)現(xiàn)丁二�；瘡V泛存在于線粒體能量代謝調(diào)控酶中,這些酶參與調(diào)控氨基酸代謝、脂肪酸代謝和三羧酸循環(huán)等多種重要的代謝通路,所以SIRT5可以通過(guò)催化去丁二酰來(lái)調(diào)節(jié)這些代謝調(diào)控酶的活性以達(dá)到調(diào)節(jié)代謝過(guò)程的目的。由于sirtuin催化的去�；磻�(yīng)與生命體的健康和疾病有著千絲萬(wàn)縷的關(guān)系,sirtuin抑制劑的開發(fā)在近幾年引起了學(xué)者們極大的興趣,這不僅會(huì)為相關(guān)疾病提供新的治療方法,也為探索sirtuin的未知世界提供了一把新的鑰匙。本課題針對(duì)SIRT6和SIRT5各設(shè)計(jì)了一系列的環(huán)肽類化合物,設(shè)計(jì)中運(yùn)用了硫脲類sirtuin抑制彈頭。主要通過(guò)固相合成的方式制得目標(biāo)化合物,將化合物分離純化后對(duì)其進(jìn)行sirtuin各亞型的體外抑制測(cè)試、蛋白酶水解穩(wěn)定性測(cè)試和免疫印跡測(cè)試。經(jīng)過(guò)一系列的綜合評(píng)估,我們發(fā)現(xiàn)在針對(duì)SIRT6設(shè)計(jì)的化合物中,含有Nε-十二烷基(或十四烷基)-硫代氨甲酰賴氨酸彈頭的環(huán)肽類化合物對(duì)SIRT6都表現(xiàn)出了強(qiáng)效的抑制活性。此外,我們還發(fā)現(xiàn)其中一個(gè)化合物雖然對(duì)SIRT1也保持著很強(qiáng)的抑制力度,但是相對(duì)于SIRT2/3/5,其仍具有較好的SIRT6抑制選擇性(分別為~20倍、~11倍和940倍)。同時(shí)我們發(fā)現(xiàn)該化合物還具備很好的蛋白酶水解穩(wěn)定性和較好的細(xì)胞通透性。在針對(duì)SIRT5設(shè)計(jì)的化合物中,我們發(fā)現(xiàn)了一個(gè)強(qiáng)效且具有選擇性的SIRT5環(huán)肽類抑制劑(相對(duì)于SIRT1/2/3/6),該化合物也被發(fā)現(xiàn)比其相應(yīng)的直鏈肽化合物具有更強(qiáng)的穩(wěn)定性。這些發(fā)展SIRT6和SIRT5抑制劑的研究為以后發(fā)展更強(qiáng)且具有選擇性、酶解穩(wěn)定且具有細(xì)胞膜通透性的代謝安全的抑制劑奠定了一個(gè)好的基礎(chǔ)。
[Abstract]:Sirtuin is a kind of 尾 -NAD dependent deacylated lysine side chain deacylase, which is not a simple amidolysis reaction, which is different from other deacylase catalyzed by acylated lysine side chain deacylation. Instead, it cleans a molecule of nicotinamide from nicotinamide adenine dinucleotides (尾 -NAD or NAD) to produce three products, the deacylated protein, There is a growing body of research that suggests that sirtuin catalyzed deacylation can regulate many important biological processes in the body. The catalytic response is also considered to be a potential target for the treatment of multiple human diseases. SIRT6 is one of the seven sirtuin SIRT1-7 found in mammals and humans, and is mainly present in the nucleus. In addition to catalytic deacetylation, SIRT6 has a stronger activity of degreasing fatty acyl (such as 14 acyl). SIRT6 can catalyze the removal of acetyl from histone H3K9 and H3K56, which regulates gene expression and maintains chromatin telomeres. In addition, SIRT6 catalyzes the removal of acetyl groups from histone H3K9 and H3K56. The deacylation reaction is also linked to many diseases, SIRT5, such as cardiovascular disease, cancer, neurodegenerative diseases and diabetes mellitus, is an enzyme that mainly exists in mitochondria, and has stronger demalonyl than catalytic deacetylation activity. Studies have found that succinylation is widely found in mitochondrial energy metabolism regulatory enzymes, these enzymes are involved in the regulation of amino acid metabolism, fatty acid metabolism and tricarboxylic acid cycle and other important metabolic pathways. Therefore, SIRT5 can regulate the activity of these metabolic regulatory enzymes by catalyzing desuccinyl. Because sirtuin catalyzed deacylation is closely related to the health and disease of life, sirtuin is closely related to the metabolism process. The development of inhibitors has aroused great interest among scholars in recent years. This will not only provide a new treatment for related diseases, but also provide a new key to explore the unknown world of sirtuin. A series of cyclic peptide compounds have been designed for SIRT6 and SIRT5 respectively. Thiourea sirtuin inhibitory warhead was used in the design. The target compounds were prepared by solid phase synthesis. The compounds were separated and purified and tested for the inhibition of sirtuin subtypes in vitro. Protease hydrolysis stability tests and Western blotting tests. After a series of comprehensive evaluations, we found that in compounds designed for SIRT6, Cyclopeptide compounds containing N 蔚 -#number0# alkyl (or 14 alkyl-carbamyl lysine warhead) showed strong inhibitory activity against SIRT6. In addition, we also found that one of the compounds maintained a strong inhibition on SIRT1. But compared with SIRT2 / 3 / 5, it still has good SIRT6 inhibition selectivity (11 times and 940 times) respectively, and we also found that the compound also has good proteolytic stability and good cell permeability. In the compounds designed for SIRT5, We have found a potent and selective SIRT5 cyclic peptide inhibitor (compared with SIRT 1 / 2 / 3 / 6), which is also found to be more stable than its corresponding straight chain peptide compounds. In the future, the development is stronger and more selective. The enzyme-stable and membrane-permeable metabolite-safe inhibitors have laid a good foundation.
【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
【分類號(hào)】：R91

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