發(fā)布時(shí)間：2018-03-07 09:31

  本文選題：艾滋病　切入點(diǎn)：HIV-1逆轉(zhuǎn)錄酶　出處：《山東大學(xué)》2014年博士論文　論文類型：學(xué)位論文

【摘要】：人免疫缺陷病毒1型(HIV-1)是艾滋病的主要病原體。自從1981年發(fā)現(xiàn)以來(lái),目前已經(jīng)成為危害人類生命健康的重大傳染性疾病。雖然高效抗逆轉(zhuǎn)錄療法(Highly Active Antiretro viral Therapy, HAART)的實(shí)施是抗艾滋病治療的一項(xiàng)重大突破,但是耐藥性的出現(xiàn)及長(zhǎng)期服藥的毒性問(wèn)題極大地限制了該療法的應(yīng)用,因此新型抗艾滋病藥物的研發(fā)依然刻不容緩。 HIV-1非核苷類逆轉(zhuǎn)錄酶抑制劑(NNRTIs)是HAART療法的重要組成部分。NNRTIs具有結(jié)構(gòu)多樣性,作用于HIV-1逆轉(zhuǎn)錄酶(Reverse Transcriptase, RT)的疏水性口袋。該類藥物具有高效低毒、特異性強(qiáng)的優(yōu)點(diǎn),然而易產(chǎn)生耐藥性的缺陷使該類藥物迅速喪失臨床效價(jià)。因此新型、高效、低毒、廣譜抗耐藥性的NNRTIs的研發(fā)是目前抗HIV藥物研究的熱點(diǎn)之一。 由于NNRTIs結(jié)合口袋(Non-nucleoside Inhibitor-Binding Pocket, NNIBP)是在NNRTIs的存在下誘導(dǎo)產(chǎn)生的,因此完全基于HIV-1RT的三維結(jié)構(gòu)進(jìn)行全新抑制劑設(shè)計(jì)還存在較大困難。故選擇研發(fā)前景較大的化合物為先導(dǎo),在對(duì)其構(gòu)效關(guān)系及結(jié)合模式分析的基礎(chǔ)上,綜合運(yùn)用結(jié)構(gòu)生物學(xué)信息、計(jì)算化學(xué)技術(shù)及傳統(tǒng)藥物化學(xué)策略進(jìn)行先導(dǎo)化合物的優(yōu)化,是當(dāng)前發(fā)現(xiàn)新一代NNRTIs藥物的有效途徑。本論文對(duì)三類HIV-1RT非核苷類逆轉(zhuǎn)錄酶抑制劑進(jìn)行了研究。 一、硝基吡啶類DAPYs非核苷類逆轉(zhuǎn)錄酶抑制劑的研究 二芳基嘧啶類衍生物(DAPYs)是一類非核苷類HIV-1逆轉(zhuǎn)錄酶抑制劑,因其高效低毒、抗耐藥性的特點(diǎn),近年來(lái)成為抗HIV藥物研發(fā)的熱點(diǎn)領(lǐng)域。2008年FDA批準(zhǔn)上市的新一代NNRTIs Etravirine(TMC125)及2011年批準(zhǔn)的Rilpivirine(TMC278)都屬于DAPY類化合物。本論文基于DAPY類逆轉(zhuǎn)錄酶抑制劑的構(gòu)效關(guān)系和DAPYs/HIV-1RT復(fù)合物晶體結(jié)構(gòu)的分析,應(yīng)用生物電子等排原理,將原本的嘧啶環(huán)替換為硝基吡啶環(huán),設(shè)計(jì)了一系列硝基吡啶類DAPY系列衍生物(7a-7r),并對(duì)其進(jìn)行了定向合成。該系列共合成了18個(gè)化合物,所有合成的化合物的結(jié)構(gòu)均經(jīng)波譜分析驗(yàn)證。 對(duì)所合成的化合物應(yīng)用MTT法進(jìn)行了體外抗野生型HIV-1(ⅢB).HIV-2(ROD)及HIV-1突變毒株的細(xì)胞活性測(cè)試。其中,化合物7b顯示出最佳的抗病毒活性(EC50=0.056μM,SI=1251).此外,化合物7k(EC50=0.034μM,SI=691),化合物7c(EC50=0.11μM,SI:339)和7h(EC50=0.17μM,SI=97)活性均比對(duì)照藥物NVP和DLV高。選取活性較好的化合物7b、7c和7k進(jìn)行了HIV-1逆轉(zhuǎn)錄酶的抑制活性測(cè)定,抑制活性分別為6.9μM.8.5μM和10.4μM,顯示出了該類化合物對(duì)HIV-1RT選擇性的抑制作用。 利用DOCk分子對(duì)接方法,將活性最好的化合物7b和7k與野生型HIV-1RT和突變型的HIV-1RT分別進(jìn)行分子模擬,并對(duì)該系列化合物與HIV-1RT的相互作用及構(gòu)效關(guān)系進(jìn)行了初步分析,為未來(lái)設(shè)計(jì)新型的高效低毒的DAPYs非核苷類逆轉(zhuǎn)錄酶抑制劑提供了有益的信息。 二、噻唑烷酮類非核苷類逆轉(zhuǎn)錄酶抑制劑的研究 噻唑烷酮類化合物是一類結(jié)構(gòu)新穎的NNRTIs,經(jīng)結(jié)構(gòu)的修飾和改造,發(fā)現(xiàn)了很多具有很高抗HIV-1活性的噻唑烷酮類化合物,具有非常廣闊的研發(fā)前景。硫代羧酰苯胺類衍生物(thiocarboxanilide)UC-781是一個(gè)非常有效的HIV-1RT抑制劑候選藥物(EC50=0.002μ.M,SI=50000).UC-781可以降低病毒顆粒的感染力,被認(rèn)為是預(yù)防HIV-1傳播的很有希望的化合物。此外,該化合物具有良好的抗耐藥譜,對(duì)多種突變毒株RT的抑制效果與對(duì)野生型HIV-1RT的接近,且可以在高劑量水平抑制NNRTIs耐藥病毒株而無(wú)任何細(xì)胞毒性。 以噻唑烷酮類NNRTIs和UC-781為先導(dǎo)物,借鑒兩類NNRTIs的結(jié)構(gòu)特征及與RT的結(jié)合模式,結(jié)合兩類化合物的相似點(diǎn),運(yùn)用分子雜合的藥物設(shè)計(jì)原理,合理地引入活性取代基,保留了先導(dǎo)化合物UC-781的異戊烯基基團(tuán),設(shè)計(jì)了一系列與靶點(diǎn)緊密結(jié)合的新型噻唑烷酮類系列化合物。并對(duì)目標(biāo)化合物進(jìn)行了定向合成,所有合成的化合物的結(jié)構(gòu)均經(jīng)波譜驗(yàn)證。應(yīng)用MTT法對(duì)所合成的化合物進(jìn)行了體外抗野生型HIV-1(ⅢB)、HIV-2(ROD)細(xì)胞活性測(cè)定。試驗(yàn)結(jié)果表明,部分噻唑烷酮類化合物表現(xiàn)出了一定的活性,但細(xì)胞毒性略高。將其中活性最好的化合物與HIV-1RT進(jìn)行分析對(duì)接,對(duì)該類化合物的構(gòu)效關(guān)系進(jìn)行了初步分析,初步闡明了該類化合物與HIV-1RT的結(jié)合位點(diǎn)與構(gòu)象,解釋活性略低的可能原因,為進(jìn)一步結(jié)構(gòu)的優(yōu)化提供新的思路。 三、DABOs類非核苷類逆轉(zhuǎn)錄酶抑制劑 二氫烷氧芐基嘧啶酮(Dihydro-alkylthio-benzyl-oxopyrimidines, DABOs)衍生物為其中較為典型的一類NNRTIs,由于其分子構(gòu)象具有柔性和結(jié)合位點(diǎn)中具有適配性的特征,可有效地抑制野生型和耐藥型病毒株的復(fù)制,成為目前抗HIV藥物研究的重要方向。 本論文基于DABO類抑制劑的構(gòu)效關(guān)系,結(jié)合計(jì)算機(jī)對(duì)接模型,依據(jù)分子雜化的藥物設(shè)計(jì)原理,以取代哌啶環(huán)來(lái)延長(zhǎng)側(cè)鏈的S-DABOs衍生物,通過(guò)變化哌啶環(huán)N原子上取代基、母環(huán)的5位、6位取代基來(lái)考察化合物各位點(diǎn)對(duì)活性的影響,并以未連有哌啶環(huán)的另一系列化合物作為對(duì)照共合成了兩個(gè)系列化合物。為了從理論上驗(yàn)證設(shè)計(jì)思想的合理性,對(duì)設(shè)計(jì)的化合物進(jìn)行了對(duì)接分析,分析結(jié)果表明目標(biāo)分子的結(jié)構(gòu)修飾具有理論上的合理性。最后根據(jù)虛擬篩選的結(jié)果,從高到低打分,確定并合成了36個(gè)目標(biāo)化合物,并選取打分較高的化合物進(jìn)行了HIV-1RT的活性測(cè)定�；钚越Y(jié)果顯示,化合物8a3-2、8a2-1和8b3-2對(duì)HIV-1RT的抑制活性分別為1.58μM、14.61μM和13.71μM,顯示出了一定的HIV-1RT抑制活性,其中在虛擬篩選中打分最高的化合物8a3-2(IC50=1.58μM)的活性高于對(duì)照藥奈韋拉平(IC5o=3.93μM),并與先導(dǎo)化合物TMC125(IC5o=1.05μM)類似,表現(xiàn)了較高的HIV-1RT抑制活性。 總之,本論文以高效抗耐藥的DAPYs、噻唑烷酮類和DABOs三類NNRTIs先導(dǎo)化合物為模板,在前人研究的基礎(chǔ)上,結(jié)合構(gòu)效關(guān)系結(jié)論及藥效團(tuán)特征,分別根據(jù)藥物設(shè)計(jì)中的生物電子等排原理和分子雜化原理,對(duì)先導(dǎo)化合物進(jìn)行了結(jié)構(gòu)多樣的骨架變換,并應(yīng)用藥物設(shè)計(jì)軟件進(jìn)行虛擬篩選�？偣苍O(shè)計(jì)合成了四個(gè)系列結(jié)構(gòu)全新的化合物。對(duì)目標(biāo)化合物進(jìn)行了抗HIV活性篩選,發(fā)現(xiàn)其中部分化合物的抗HIV-1(IIIB)活性達(dá)到或超過(guò)上市藥物奈韋拉平與地拉韋定,具有進(jìn)一步研究與開(kāi)發(fā)價(jià)值。此外,還對(duì)目標(biāo)化合物進(jìn)行了計(jì)算機(jī)分子對(duì)接研究,根據(jù)其與HIV-1RT的結(jié)合模式對(duì)化合物的構(gòu)效關(guān)系進(jìn)行了初步分析,為進(jìn)一步的研究提供了有利的信息。
[Abstract]:Human immunodeficiency virus type 1 (HIV-1) is the main pathogen of AIDS. Since discovered in 1981, has become the major infectious diseases endangering human life and health. Although effective antiretroviral therapy (Highly Active Antiretro viral Therapy, HAART) is a major breakthrough in the implementation of anti HIV treatment, but the emergence of drug resistance and toxicity problems the long-term medication greatly limits the application of the therapy, so the development of the new anti AIDS drugs is still urgent.
HIV-1 non nucleoside reverse transcriptase inhibitor (NNRTIs) is an important part of HAART therapy.NNRTIs has structural diversity, in the role of HIV-1 reverse transcriptase (Reverse Transcriptase, RT) of the hydrophobic pocket. The drugs with high efficiency and low toxicity, strong specificity, but drug-resistant defects make the drugs rapidly lost clinical value of model. Therefore, high efficiency, low toxicity, broad-spectrum drug resistance research of NNRTIs is one of the hot study of anti HIV drugs at present.
Due to the NNRTIs binding pocket (Non-nucleoside Inhibitor-Binding Pocket, NNIBP) is induced in the presence of NNRTIs, thus complete the three-dimensional structure of HIV-1RT based on a new inhibitor design still has many difficulties. The choice of R & D promising compounds as first, in the structure-activity relationship and based on the analysis of the model, the integrated use of the structure of biological information, optimization strategy of chemical technology and traditional medicine chemical compounds, is an effective way for a new generation of NNRTIs drug discovery at present. This paper focuses on three classes of HIV-1RT non nucleoside reverse transcriptase inhibitors were studied.
A study of nitropyridine DAPYs non nucleoside reverse transcriptase inhibitors
Two aryl pyrimidine derivatives (DAPYs) is a kind of HIV-1 non nucleoside reverse transcriptase inhibitors, because of its high efficiency and low toxicity, anti drug, anti HIV drugs in recent years become a hot spot in research field of.2008 FDA approved the listing of the new generation of NNRTIs Etravirine (TMC125) Rilpivirine and approved in 2011 (TMC278) belong to DAPY compounds. This paper analysis of structure effect relationship and DAPYs/HIV-1RT crystal structures of complexes of DAPY reverse transcriptase inhibitors based on the application of the principle of bioisosterism, originally a pyrimidine ring substituted for nitro pyridine ring derivatives, a series of nitro pyridine DAPY series design (7a-7r), and has carried on the synthesis. This series of 18 compounds were synthesized, all structures of the synthesized compounds were confirmed by spectral analysis.
The application of MTT compound prepared by the method of in vitro anti wild type HIV-1 (B III).HIV-2 (ROD) and HIV-1 mutant strain cell activity test. Among them, compound 7b shows the best antiviral activity (EC50=0.056 M, SI=1251). In addition, the compound 7K (EC50=0.034 M, SI=691, 7C) compounds (EC50=0.11 M, SI:339) and 7h (EC50=0.17 M, SI=97) activity was higher than that of the control drugs NVP and DLV. Choose good activity compounds 7b, 7C and 7K were determined by reverse transcriptase HIV-1 inhibitory activity, inhibitory activity of 6.9 M.8.5 M and 10.4 M, the inhibitory effect of the compounds on the selectivity of HIV-1RT display.
Using DOCk molecular docking method, the best activity of compound 7b and 7K with wild-type HIV-1RT and mutant HIV-1RT were molecular simulation, and the interaction of these compounds with HIV-1RT and structure-activity relationship were analyzed, provide useful information for the future design of high efficiency and low toxicity of new non nucleoside DAPYs reverse transcriptase inhibitors.
Two, a study of thiazolidone non nucleoside reverse transcriptase inhibitors
Thiazolidinones are a kind of novel structure of NNRTIs, the modification of the structure, found a lot of high anti HIV-1 activity of thiazolidinones, has very broad prospects for development. Thio carboxylic acylaniline derivative (thiocarboxanilide) UC-781 is a very effective drug candidate (HIV-1RT inhibitor EC50=0.002.M, SI=50000).UC-781 can reduce the infectious virus particles, considered compounds to prevent the spread of HIV-1 is very promising. In addition, the compound has good anti drug resistance, inhibition effect on a variety of mutant strain RT and the close of wild type HIV-1RT, and can inhibit the NNRTIs resistance at high dose levels virus without any cytotoxicity.
The thiazolidinone class NNRTIs and UC-781 as the lead compound, combined with the structure characteristics of two types of reference mode of NNRTIs and RT, with two kinds of compounds are similar, using the principle of molecular drug design heterozygous, reasonably introducing reactive substituents, retains the prenyl groups of lead compounds of UC-781, combined with a series of with the target of new thiazolidinones compound series is designed. And the target compounds were directed synthesis, structure of all the synthesized compounds were confirmed by spectral verification. By applying the MTT method against wild type HIV-1 in vitro of the synthesized compounds (B III), HIV-2 (ROD) cell activity assay. The test results that part of thiazolidinone compounds exhibited certain activity, but slightly higher. The cytotoxicity of compounds with HIV-1RT activity were analyzed for the best docking, the structure-activity relationship of these compounds were preliminary The binding sites and conformations of these compounds with HIV-1RT were preliminarily elucidated, and possible reasons for their low activity were explained, providing new ideas for further structural optimization.
Three, DABOs non nucleoside reverse transcriptase inhibitors
Two hydrogen alkoxy benzyl pyrimidine ketone derivatives (Dihydro-alkylthio-benzyl-oxopyrimidines, DABOs) for a class of NNRTIs which is more typical, because of its molecular conformation and binding sites with flexible adaptation features, can effectively inhibit the wild-type and drug-resistant strains of the virus replication, has become an important direction of the present study of anti HIV drugs.
The structure-activity relationship of DABO inhibitors based on the combination of computer docking model, based on the principle of drug design, molecular hybrid, S-DABOs derivatives to replace the piperidine ring to extend side chain, through the change of the piperidine ring substituent on the N atom, 5 ring, 6 substituents to examine all compounds on the activity the influence and not even a series of compound piperidine ring as the control of the two series of compounds were synthesized. In order to verify the rationality of design ideas from the theory, to design the compound for the docking analysis, structural modification analysis result of standard molecular eyesight have rationality in theory. According to virtual screening the results, from high to low scoring, and determine the 36 target compounds were synthesized, and selected the high scoring compounds were determined by HIV-1RT activity. The activity results showed that compounds 8a3-2,8a2-1 and 8b3-2 on H The inhibitory activity of IV-1RT were 1.58 M, 14.61 M and 13.71 M, showing a certain HIV-1RT inhibitory activity, which in the virtual screening scoring highest compound 8a3-2 (IC50=1.58 M) activity was higher than that of the control Po Nai Vee Lapin (IC5o=3.93 M), and TMC125 (IC5o=1.05 M) lead compounds similar that showed higher HIV-1RT inhibitory activity.
In a word, in this dissertation, resistance DAPYs, NNRTIs lead compounds of thiazolidinones and DABOs three as the template, on the basis of previous studies, combined with the conclusion of structure-activity relationship and pharmacophore features, respectively, according to the principle of bioisosterism and molecular hybridization principle in drug design, the framework of lead compound transform structure diversity, and virtual screening of drug design software. A total of four series of design structure of the new compounds were synthesized. The target compounds were screened for anti HIV activity, found that some of them were anti HIV-1 (IIIB) activity reached or exceeded the listed drug nevirapine and delavirdine, with further research and the development of value. In addition, the target compounds were studied by computer molecular docking, according to the binding mode with HIV-1RT on their structure-activity relationship were analyzed for Further research provides favorable information.

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R96;R914.5

【共引文獻(xiàn)】

相關(guān)期刊論文 前4條

1 孫玉娜;王浩;薛友林;李輝;宋有濤;;Ydj1p C端二聚體中螺旋α3與domain Ⅲ分離的拉伸分子動(dòng)力學(xué)模擬[J];生物物理學(xué)報(bào);2014年01期

2 ;The development of anti-HIV-1 drugs[J];藥學(xué)學(xué)報(bào);2010年02期

3 陳華;黃長(zhǎng)軍;朱墨;李小六;;新型含親水基的噻唑烷-4-酮衍生物的合成及HIV逆轉(zhuǎn)錄酶抑制活性[J];有機(jī)化學(xué);2014年04期

4 古雙喜;喬恒;段婷;朱園園;陳金芳;巨修練;;HIV-1抑制劑依曲韋林的合成[J];武漢工程大學(xué)學(xué)報(bào);2014年06期

相關(guān)會(huì)議論文 前1條

1 黃長(zhǎng)軍;朱墨;陳華;李小六;;新型1,3-噻唑烷-4-酮衍生物的合成及抗HIV-RT活性評(píng)價(jià)[A];中國(guó)化學(xué)會(huì)第29屆學(xué)術(shù)年會(huì)摘要集——第07分會(huì)：有機(jī)化學(xué)[C];2014年

相關(guān)博士學(xué)位論文 前6條

1 展鵬;新型芳唑（嗪）巰乙酰胺類HIV-1 NNRTI的設(shè)計(jì)、合成與活性研究[D];山東大學(xué);2010年

2 倪小菊;HIV整合酶對(duì)鏈轉(zhuǎn)移抑制劑的易感性和耐藥性研究以及新型抗整合酶單域抗體的研發(fā)[D];華東師范大學(xué);2011年

3 張芬芬;抗菌類藥物電化學(xué)傳感器的構(gòu)建及性能研究[D];上海大學(xué);2013年

4 馬靜;HIV對(duì)逆轉(zhuǎn)錄酶抑制劑2’，3’-雙脫氫-3’-脫氧-4’-乙炔基胸苷（114）耐藥機(jī)制的研究[D];中南大學(xué);2013年

5 宋永彬;新型氧雜蒽和呋喃衍生物的合成及其抗腫瘤活性研究[D];哈爾濱理工大學(xué);2013年

6 陳緒旺;新型哌啶取代DAPY類HIV-1非核苷逆轉(zhuǎn)錄酶抑制劑的設(shè)計(jì)、合成及活性研究[D];山東大學(xué);2014年

相關(guān)碩士學(xué)位論文 前3條

1 肖富貴;植物非光化學(xué)能量耗散機(jī)制的理論研究[D];山東理工大學(xué);2012年

2 劉慧杰;DATA類化合物抗HIV-1活性的理論研究[D];南華大學(xué);2013年

3 欒悅;拉伸分子動(dòng)力學(xué)模擬研究Hsp31和底物的相互作用[D];吉林大學(xué);2014年

，

本文編號(hào)：1578864