新型NNRTIs二芳基苯胺类和HIV成熟抑制剂Bevirimat双靶点化合物的设计、合成、生物活性及体外代谢评价

发布时间：2018-01-13 19:33

本文关键词：新型NNRTIs二芳基苯胺类和HIV成熟抑制剂Bevirimat双靶点化合物的设计、合成、生物活性及体外代谢评价　出处：《中国人民解放军军事医学科学院》2017年硕士论文　论文类型：学位论文

【摘要】：艾滋病(acquired immunodeficiency syndrome,AIDS)是由人类免疫缺陷病毒(human immunodeficiency virus,HIV)感染引起的疾病,也是当前医学界难以治愈的顽疾之一。2016年全球艾滋病统计数据估计,2015年有3670万人感染艾滋病毒。其中,200万人是新增感染者,110万人死于与艾滋病有关的疾病。尽管针对病毒生命的不同阶段进行抑制的被批准上市的药物已有30多种,但还没有彻底治愈HIV感染的疗法。临床上使用的高效抗逆转录病毒疗法能有效缓解患者的病情,极大的降低患者体内的病毒载量,延长患者寿命,但该疗法并不能从根源上将患者体内的HIV病毒清除,还存在明显毒副作用,患者依从性较差,研究新型抗HIV药物为临床提供安全,经济用药选择变得尤为重要。本课题组前期研究发现两类新结构类型的非核苷类逆转录酶抑制剂(non-nucleoside reverse transcriptase inhibitors,NNRTIs)先导化合物:二芳基吡啶胺类(diarylpyridinamines,DAPAs)和二芳基苯胺类(diarylanilines,DAANs)化合物,它们对HIV野生型和多种耐药型病毒均有较强抑制活性,与同类上市药Riplivirine的活性相当,并能有效抑制其新生耐药性病毒的复制。Bevirimat(BVM,3-O-(3′,3′-dimethylsuccinyl)betulinic acid)是第一个进入临床试验的成熟抑制剂。它干扰了CA-SP1(capsid-spacer protein 1)到CA(capsid)的加工,导致CA-SP1的积累并组装释放未成熟的HIV病毒,此病毒不具有感染能力,从而阻断了HIV病毒的传播。已通过IIa期临床试验,证明了其临床疗效和安全性。作为持续性研究,本课题意将高活性的二芳基苯胺类化合物与bevirimat经适当的中间体连接,合成双靶点化合物,再对其进行活性以及体外代谢性能的考察。以期发现高活性、体内易代谢分离、可同时作用于不同靶点的双靶点新型化合物,发挥1+12的协同抗HIV的作用和克服HIV药物的耐药性问题。本文对中间体和偶合方法进行了探索,将不同类型的中间体连在DAAN母核B环的2-或4-位上,共计合成20个二芳基苯胺类衍生物,并对新合成的二芳基苯胺类衍生物进行了抗HIV病毒活性和代谢稳定性评价,其中活性化合物16个,经活性及类药性评价得构效关系如下:1)在DAAN的B环4-位进行连接侧链,取代基R2较长或者连接基团极性增大,都会使活性降低;引入短链可以保持或者改善分子的抗HIV活性,在分子抗耐药方面的表现短链优于长链。2)在人肝微粒体模型代谢稳定性试验当中,二芳基苯胺类化合物中B环2-位酰胺基取代的化合物比B环4-位酰胺取代的化合物更易不稳定,且B环4-位酯键修饰要比酰胺键侧链更不稳定。3)在DAAN类中B环2-位氨基上连接脂肪链R1后,均会减弱DAAN母核的抗HIV活性,游离出氨基,抗HIV活性最好,这也与前其研究结论一致,此位点与K101的氨基酸残基能产生作用,是抗HIV病毒的活性关键基团。经探索合成3个DAAN与BVM双靶点偶合物,并没有检测到抗HIV活性,对中间体的尝试还有待提高。由人肝微粒体代谢稳定性试验可知,双靶点化合物的代谢半衰期t1/2均明显小于参比对照盐酸普奈洛尔和TMC278,说明双靶点化合物能够在酶的参与下发生代谢。相对应的DAAN中间体也进行了代谢稳定性考察,代谢半衰期t1/2分别为10.7 min,75.3 min,117.5 min。暗示在DAAN类的B-环2-位的氨基连接侧链的长度越长,越容易被代谢,可以指导中间体的进一步探索。综上所述,本课题经探索合成了非核苷类逆转录酶抑制剂与HIV成熟抑制剂BVM的双靶点化合物,且在肝微粒体酶作用下易代谢,为后期对其活性的优化奠定了基础。
[Abstract]:AIDS (acquired immunodeficiency syndrome, AIDS) is by the human immunodeficiency virus (human immunodeficiency, virus, HIV) infection caused by disease, but also the medical profession is difficult to cure the ills of one of the.2016 global AIDS statistics estimated that in 2015 there are 36 million 700 thousand people infected with HIV. Among them, 2 million people are newly infected, 1 million 100 thousand people died of AIDS related the disease. Although there are different stages of life for the virus inhibition of approved drugs 30 kinds, but not completely cure HIV infection therapy. Can effectively relieve patients with highly active antiretroviral therapy in clinical use, reduce the patient's viral load greatly, prolong the life of patients, but the not from HIV therapy and viral clearance in patients on the root, there are obvious side effects, patients with poor compliance, research model Anti HIV drugs provide safety for clinical drug selection, the economy has become particularly important. Our previous study found that non nucleoside reverse transcriptase inhibitors of two kinds of new structure type (non-nucleoside reverse transcriptase inhibitors, NNRTIs) compounds: two aryl amine pyridine (diarylpyridinamines, DAPAs) and two aryl aniline (diarylanilines, DAANs) the compound, HIV wild type and multi drug resistant virus had strong inhibitory activity, and a similar listed drug Riplivirine activity,.Bevirimat replication and can effectively inhibit the newborn of drug-resistant virus (BVM, 3-O- (3 ', 3' -dimethylsuccinyl) betulinic acid) is the first mature inhibitor to enter clinical trials. It is interference CA-SP1 (capsid-spacer protein 1) to CA (capsid) processing, and lead to the accumulation of CA-SP1 assembly release was not mature HIV virus, this virus is not out Have the infection ability, thereby blocking the spread of the HIV virus. Through phase IIa clinical trials to prove its clinical efficacy and safety. As the research continues, this topic will high activity of two aryl aniline compounds bevirimat with appropriate intermediate connection, synthesis of two target compounds, and study activity and in vitro metabolism on the performance of the high activity. In order to find the body, easy metabolism separation, new double target on different targets can also play a role of synergistic anti 1+12 compounds, the role of HIV and HIV drugs to overcome the resistance problem. This paper explores the intermediate and intermediate coupling method, the different types of even in the DAAN nucleus of B ring 2- or 4-, a total synthesis of 20 two aryl aniline derivatives, and two aryl aniline derivatives newly synthesized by anti HIV virus activity and metabolic stability evaluation, including live Of the 16 compounds, the activity and structure-activity relationship of drug like evaluation was as follows: 1) the connection side chain in the DAAN B ring 4-, R2 connection or longer substituent group polarity increases, will reduce the activity; introduction of short chain molecules can maintain or improve the anti HIV activity, short chain performance better than the long chain.2 in the sub resistance) in human liver microsomes model metabolic stability test, compounds B ring 2- two aryl amide substituted aniline compounds than in the B ring 4- amide substituted more unstable, and the B ring 4- ester bond modification than amide bond the side chain is more unstable.3) in the DAAN class B ring 2- amino fatty chain connected to R1, DAAN will decrease the parent nucleus of anti HIV activity, free amino group, the best anti HIV activity, and it also before the conclusion of the study, the amino acid residues in this site and K101 could produce. Anti HIV virus activity The key to explore the synthesis of the 3 groups. The DAAN and BVM double target conjugates, and no detectable anti HIV activity, try to intermediate still needs to be improved. The experiment result shows the metabolic stability in human liver microsomes, double target compounds metabolism half-life of t1/2 were obviously lower than the reference control salt Um F Naylor and TMC278, show that the double the target compounds can occur in metabolic enzymes. DAAN corresponding intermediates were also investigated metabolic stability, metabolic half-life of t1/2 was 10.7 min, 75.3 min, 117.5 min., suggesting a connection side chain in the DAAN class B- 2- amino ring length is longer, the easier it is to further explore metabolism. Guidance for intermediates. In summary, this topic through the study on the synthesis of double targeting non nucleoside reverse transcriptase inhibitor and HIV inhibitor BVM mature compounds, and in liver microsomal enzymes under the action of the latter is easy to metabolism. The optimization of activity lays the foundation.

【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R914;R96

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