發(fā)布時間：2018-05-13 04:40

  本文選題：艾滋病 + 人類免疫缺陷病毒�。� 參考：《北京工業(yè)大學(xué)》2014年碩士論文

【摘要】：自1981年美國研究人員發(fā)現(xiàn)世界首例AIDS病例后，AIDS在全球范圍內(nèi)迅速蔓延,且至今尚無有效的治療方法。它嚴(yán)重的威脅著人類的公共衛(wèi)生安全。AIDS由人類免疫缺陷病毒引起，HIV包括兩種密切相關(guān)的類型，分別為HIV-1和HIV-2，絕大多數(shù)AIDS是由HIV-1引起的。 HIV-1的復(fù)制要經(jīng)過穿入、逆轉(zhuǎn)錄、整合、基因表達(dá)、裝配、出芽和成熟等一系列步驟。其中HIV-1的整合酶在病毒的復(fù)制過程中起著十分重要的作用，它可將反轉(zhuǎn)錄得到的雙鏈DNA插入宿主細(xì)胞基因當(dāng)中，幫助病毒DNA與宿主DNA的融合。最后在各種病毒非結(jié)構(gòu)蛋白及細(xì)胞蛋白的參與下完成病毒顆粒的復(fù)制。整合酶介導(dǎo)著病毒基因與宿主基因之間的融合，是尋找抗HIV藥物的理想靶點。 本研究的主要目的是通過一系列的研究手段從天然藥物KA-60中篩選出有效地抗HIV-1整合酶單體。實驗在制備高純度HIV-1整合酶的基礎(chǔ)上，，對天然藥物KA-60進(jìn)行提取，以整合酶為靶點利用SPR技術(shù)對所提取部位進(jìn)行篩選，得到有效的藥物提取部位。接著對所提取部位進(jìn)行細(xì)胞內(nèi)藥效學(xué)驗證。利用一種蛋白偶聯(lián)柱偶聯(lián)整合酶蛋白對所提取藥物進(jìn)行分離提純，得到富集后的產(chǎn)物KA-60-6與KA-60-7。然后借助HPLC-MS對分離得到的富集產(chǎn)物進(jìn)行分析，得到它們的準(zhǔn)分子量并與化合物分子量數(shù)據(jù)庫進(jìn)行比對，篩選出藥物中分子量與之相對應(yīng)的物質(zhì)H、A、E，并對其進(jìn)行細(xì)胞內(nèi)藥效學(xué)檢測，確定其抗HIV-1的效果。并輔以MacSynergy II軟件進(jìn)行有效成分聯(lián)合用藥水平的檢測。最終，利用Autodock軟件模擬有效成分與HIV-1整合酶的對接，結(jié)果顯示三者均與HIV-1整合酶有一定的結(jié)合，其中單體H的結(jié)合能力最強(qiáng)，與藥效學(xué)實驗結(jié)果相符，也為之后藥物結(jié)構(gòu)的優(yōu)化與改造奠定了基礎(chǔ)。
[Abstract]:Since the United States researchers found the world's first case of AIDS cases in 1981, AIDS has spread rapidly around the world, and there is no effective treatment yet. It seriously threatens human public health security.AIDS caused by human immunodeficiency virus, and HIV includes two closely related types, HIV-1 and HIV-2, and the vast majority of AIDS are Caused by HIV-1.
HIV-1 replication has to pass through a series of steps such as penetration, reverse transcription, integration, gene expression, assembly, buds and maturation. In which HIV-1 integrase plays a very important role in the replication of the virus, it can insert reverse transcriptional double stranded DNA into the host cell gene and help the fusion of virus DNA and host DNA. The virus particles are replicated with the participation of non structural proteins and cell proteins. Integrase mediated fusion between the virus gene and the host gene is an ideal target for the search for anti HIV drugs.
The main purpose of this study is to screen out effective anti HIV-1 integrase monomers from natural drug KA-60 through a series of research methods. On the basis of the preparation of high purity HIV-1 integrase, the extraction of natural drug KA-60 was carried out, and the target sites were screened by SPR technology with integrated enzyme as the target, and effective drug extraction was obtained. The extracted parts were tested in cell pharmacodynamics. The extracted drugs were separated and purified with a protein coupling column coupled integrase protein. The enriched products, KA-60-6 and KA-60-7., were then analyzed with the aid of HPLC-MS, and their molecular weights and compounds were obtained. The sub quantity database was compared, and the substances corresponding to the molecular weight of the drug H, A, and E were screened out, and the intracellular pharmacodynamic test was carried out to determine the effect of its anti HIV-1. And the MacSynergy II software was used to test the level of the combined use of effective components. Finally, the Autodock software was used to simulate the docking of the effective components with the HIV-1 integrase. The results showed that both of the three were combined with the HIV-1 integrase, and the binding ability of the monomer H was the strongest, which was consistent with the results of the pharmacodynamics experiment. It also laid the foundation for the optimization and transformation of the drug structure.

【學(xué)位授予單位】：北京工業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R96

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相關(guān)期刊論文 前3條

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3 沈?qū)W彬;葉劍;楊立莉;陳歡;樓宜嘉;Samson A.Chow;;HIV-1整合酶的功能及其抑制劑的研究進(jìn)展[J];中國細(xì)胞生物學(xué)學(xué)報;2013年10期

本文編號：1881725