新型小分子化合物用于激活HIV潜伏病毒储存库的研究

发布时间：2018-08-26 14:12

【摘要】：人类获得性免疫缺陷综合征(AIDS),自1981年发现至今已有30多年,仍在严重威胁人类的健康。高效抗逆转录药物疗法(HAART)的发展有效控制了HIV患者病情的进展,使得HIV成为一种慢性病,一旦中断HAART治疗后,病毒会很快反弹,因此病人不得不终生服药。HIV难以治愈的主要原因是细胞中潜伏存在的病毒储存库。清除病毒储存库最有希望的策略之一是shock and kill,即先用药物激活潜伏HIV储存库的转录表达,然后利用机体免疫系统或细胞病变效应杀死这些激活后的感染细胞。目前已有多种类型的HIV病毒储存库激活剂正在研究开发中,例如IL-2、CD3单克隆抗体等会引起普遍的T细胞激活现象,但同时引起细胞毒性并且释放炎性因子;组蛋白去乙酰化酶抑制剂(HDACis)丙戊酸钠(VPA)和环庚烷异羟肟酸(SAHA)也已被证明在体外可高效诱导HIV转录的,但其临床治疗效果不佳。因此研发新型的小分子化合物用以高效安全地激活HIV潜伏病毒储存库是非常有必要的。吡喃并吲哚类化合物被广泛用于药物研发,它已被证明具有抗炎、抗肿瘤、抗细菌感染等生物活性。在先前的工作中,我们的合作者发展了一种高效合成二氢吡喃吲哚衍生物的有机催化合成方法,得到了一系列吡喃并吲哚类化合物。在本研究中,我们对这些化合物可能的生物活性进行了研究。首先使用细胞模型—J-lat A10.6细胞系,对这些化合物进行了筛选以发现可能的病毒储存库激活剂。通过细胞模型筛选实验以及化合物结构优化改造,我们最终证实化合物GIBH-LRA002 (2-氨基-3-氰基-9-甲基-4-(三氟甲基)-4,9-二氢吡喃并[2,3-b]吲哚-4-羧酸乙酯)具有最优的激活效果。由于细胞系潜伏感染模型并不能很好地代表艾滋感染者体内的病毒储存库情况,因此我们进一步验证了该化合物对SIV感染猕猴和HIV患者体内CD4+T细胞中潜伏病毒储存库的激活效果,研究结果表明了GIBH-LRA002可有效的激活这些原代CD4+T细胞中潜伏病毒储存库。此外,我们的研究也发现GIBH-LRA002对细胞活性没有显著影响。还初步证明了GIBH-LRA002不会导致T细胞的非特异活化,这将有利于阻止CD4+T细胞的过度活化,从而可降低病毒储存库激活后产生的新生病毒颗粒的扩散。总之,本研究表明了 GIBH-LRA002对HIV潜伏病毒储存库有良好的激活效果,且不会引起明显的细胞毒性和非特异激活,是一个值得深入研究的HIV病毒储存库激活候选药物。本论为将对HIV的治愈或功能治愈提供重要参考。
[Abstract]:Human acquired immunodeficiency syndrome (AIDS),) has been discovered for more than 30 years since its discovery in 1981 and is still a serious threat to human health. The development of (HAART) has effectively controlled the progress of patients with HIV, making HIV a chronic disease. Once HAART treatment is interrupted, the virus bounces back quickly. The main reason why patients have to take drugs for life. HIV is hard to cure is the virus repository that lurks in the cells. One of the most promising strategies for clearing viral repositories is that shock and kill, is first used to activate the transcription expression of latent HIV repositories and then to kill these activated infected cells by using the immune system or cytopathic effects. At present, there are many kinds of HIV virus library activators, such as IL-2,CD3 monoclonal antibody, which can cause T cell activation, but also cause cytotoxicity and release inflammatory factors. Histone deacetylase inhibitor (HDACis) valproate sodium (VPA) and cycloheptane hydroxamic acid (SAHA) have also been shown to efficiently induce HIV transcription in vitro, but its clinical efficacy is poor. Therefore, it is necessary to develop new small molecular compounds to efficiently and safely activate the HIV latent virus repository. Pyran-indoles have been widely used in drug research and development, and have been proved to have anti-inflammatory, anti-tumor, anti-bacterial infection and other biological activities. In our previous work, our collaborators developed an organic catalytic synthesis method for the synthesis of dihydropyranindoles with high efficiency and obtained a series of pyranoindole derivatives. In this study, we studied the possible biological activities of these compounds. First, the cell model of -J-lat A10.6 cell line was used to screen these compounds to identify possible viral repository activators. Through the cell model screening experiment and the structure optimization modification of the compound, we finally confirmed that the compound GIBH-LRA002 (2-amino-3-cyano-9-methyl-4- (trifluoromethyl) -4-dihydropyrromoxylic acid) has the best activation effect. Because the model of latent infection of cell line does not represent the virus repository in HIV infected person, Therefore, we further verified the activation effect of this compound on CD4 T cells in CD4 T cells of SIV infected rhesus monkeys and HIV patients. The results showed that GIBH-LRA002 could effectively activate these latent virus repositories in primary CD4 T cells. In addition, our study also found that GIBH-LRA002 has no significant effect on cell activity. It was also preliminarily proved that GIBH-LRA002 does not lead to the non-specific activation of T cells, which will help to prevent the excessive activation of CD4 T cells, and thus reduce the proliferation of new virus particles generated by the activation of virus repositories. All in all, this study shows that GIBH-LRA002 has a good activation effect on HIV latent virus repository and does not cause obvious cytotoxicity and non-specific activation. It is a promising candidate for activation of HIV virus repository. This theory provides an important reference for the cure or functional cure of HIV.
【学位授予单位】：中国科学技术大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R512.91

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