HIV整合酶抑制剂体外筛选方法研究进展

为了评价HIV整合酶抑制剂体外筛选方法的特点和整合酶抑制剂研究进展，以常用的3’加工反应法、链转移反应法、改良ELISA法及基于结构的筛选为基础归类整合酶抑制剂。整合酶抑制剂按方法分为五类，并可推测其作用机制，通过比较可知以上方法各有利弊，需配合运用。整合酶抑制剂有研究价值。     

HIV-1整合酶抑制剂的研究进展

HIV-1整合酶(integrase)是逆转录病毒复制所必需的酶,因而成为抗艾滋病(AIDS)药物设计的一个合理的靶点。本文综述了近几年的HIV-1整合酶及其抑制剂的发展现状,就如何将作用于整合酶靶点的先导化合物转变成有效的抗艾滋病药物进行了讨论。     

一种可用于天然产物粗提物的HIV-1逆转录酶抑制剂筛选方法

人类免疫缺陷病毒（HIV）逆转录酶是成功的获得性免疫缺陷综合症（AIDS）治疗药靶，HIV逆转录酶抑制剂是AIDS治疗中必不可少的药物。现有药物的副作用和耐药性的快速出现促使加快寻找新HIV逆转录酶抑制剂的研究工作。天然产物是新药的重要来源，如能够对天然产物粗提物进行HIV逆转录酶抑制活性筛选将极大地提高发现新HIV逆转录酶抑制剂的效率，但目前还未见可用于天然产物粗提物的HIV逆转录酶抑制活性的筛选方法。本文依据HIV逆转录酶以RNA为模板催化DNA合成的原理，利用biotin-dUTP和digoxigenin-dUTP在逆转录反应过程中随机参入DNA，形成digoxigenin和biotin双标记的DNA，借助digoxigenin-anti-digoxigenin系统捕获digoxigenin标记DNA，再通过生物素．链霉亲和素系统将铕螯合物标记在DNA上，以时间分辨模式测定结合的铕螫合物的荧光强度，得到HIV-1逆转录酶的活性，该方法可直接用于天然产物粗提物的HIV逆转录酶抑制活性的筛选，而不受内源性生物素的干扰。     

HIV-1衣壳蛋白抑制剂体外筛选方法研究进展

HIV-1的感染高度依赖其圆锥形的"富勒烯"型衣壳。衣壳由大约1 500个衣壳蛋白组装而成。近年来,衣壳蛋白已成为设计和筛选抗HIV-1药物的理想靶点。目前,文献已报道了多种基于不同原理的HIV-1衣壳蛋白抑制剂的筛选方法,该文将对这些体外筛选方法进行综述。     

HIV-1整合酶抑制剂的研究进展

HIV-1整合酶是设计抗艾滋病药物的理想靶点，综述了HIV-1整合酶抑制剂的研究进展。     

HIV-1整合酶3'端加工抑制剂筛选方法的建立与优化

目的建立与优化HIV-1整合酶3'端加工抑制剂筛选方法。方法利用荧光共振能量转移原理建立筛选方法,用DNaseⅠ切割底物DNA确定底物检测波长,在该检测波长下,对缓冲液成分、底物浓度、酶浓度、金属离子浓度等影响整合酶活性的条件进行优化,并用阳性药物雷特格韦和杨梅黄素进行方法验证。结果检测波长为495 nm/525 nm,使用缓冲液1、底物浓度500 nmol·L-1,整合酶浓度1μmol·L-1,镁离子浓度20 mmol·L-1时整合酶3'端加工活性最强。在此反应条件下雷特格韦和杨梅黄素能有效地抑制整合酶3'端加工活性。用所建立的方法筛选到2个有较强抑制整合酶3'端加工活性的抑制剂。结论该文成功建立并优化了HIV-1整合酶3'端加工抑制剂筛选方法。     

两种方法检测HBV标志物阳性患者血清HBV DNA载量结果

目的:观察不同方法检测乙型肝炎病毒(HBV)感染者血清中HBVDNA载量的相关性和临床意义。方法:采用实时荧光定量PCR(RQ-PCR)和核酸扩增(PCR)酶联免疫吸附(ELISA)两种方法检测HBV感染者血清HBVDNA载量。结果:HBsAg阳性各种模式两种方法检测结果比较,差异均有显著性(P<0.001)。模式Ⅴ(抗-HBs阳性+抗-HBc阳性+抗-HBe阳性)RQ-PCR检测结果<3.0lg拷贝/ml,PCR-ELISA检测结果为(3.80±1.61)lg拷贝/ml。模式Ⅵ(抗-HBc阳性+抗-HBe阳性)、Ⅶ(抗-HBc阳性)、Ⅷ(抗-HBe阳性)两种方法检测结果均﹤3.0lg拷贝/ml。结论:采用不同的方法检测HBV感染者血清中HBVDNA载量,结果存在不同程度的差异。     

基于结构的HIV-1整合酶抑制剂设计:计算机模拟方法

HIV整合酶是一种病毒编码蛋白质，它催化病毒DNA整合进入宿主基因组，这为开发新的抗HIV和抗艾滋病疗法提供了一个重要的靶标。综述通过计算机模拟方法进行基于结构的：HIV-1整合酶抑制剂设计，内容包括基于配体(如药效团)和基于靶向(如对接)的设计方法以及三维定量构效关系研究。     

二酮酸类HIV-1整合酶抑制剂的研究进展

HIV-1整合酶催化前病毒DNA整合进入宿主细胞基因组的过程是病毒复制必不可少的步骤,所以抑制HIV-1整合酶活性是治疗HIV-1感染的合理策略。目前已开发了大量HIV-1整合酶抑制剂,其中一些已进入临床研究阶段,从化学结构看,绝大多数已报道的HIV-1整合酶抑制剂属于二酮酸类或其生物电子等排体。该文对近3年来二酮酸类HIV-1整合酶抑制剂的研究进展做简要综述。     

Advances in two-metal chelation inhibitors of HIV integrase

Background: Small-molecule inhibitors of the retroviral enzyme HIV integrase are among the most exciting discoveries of the past 10 years in the area of infectious disease research. The field of two-metal chelating inhibitors of this nucleic-acid-modifying enzyme has exploded since the early diketo-acid discoveries in the late 1990s and the advent of the more drug-like heterocyclic templates in the early 2000s. The field of research has now expanded across multiple companies, resulting in several clinical candidates, and ultimately delivered the first licensed inhibitor – raltegravir – in late 2007. Objective: This review is an attempt to assess the most relevant patent literature concerning two-metal chelators from the pioneering examples in 1999 that first displayed antiviral activity through mid-2008. Methods: The existing integrase inhibitor patent literature was surveyed for chemical series that consisted of a two-metal binding pharmacaphore, which were further broken down into structural subclasses for presentation. Results/conclusions: The integrase inhibitor field has matured rapidly over the past decade into a major area of focus for new HIV therapeutics. This work has resulted in one licensed drug and clearly defined needs for future directions centered around viral resistance, for which solutions that are contained within the reviewed patents will undoubtedly emerge.     

The clinical pharmacology of integrase inhibitors

Introduction: Treatment of HIV infection has consistently evolved in the last three decades. A steady improvement in efficacy tolerability, safety, and practical aspects of treatment intake has made HIV infection much easier to manage over the long term, and in optimal treatment conditions the life expectancy of persons living with HIV infection now approaches the values of the general population. The last category of antiretrovirals to be fully developed for clinical use is the one of strand-transfer integrase inhibitors (INSTIs).

Areas covered: In this review, the evolution of the knowledge on INSTIs use in the clinical setting is reviewed, analyzed, and interpreted. Emphasis is placed on the properties possibly accounting for several superiority results achieved by INSTIs in non-inferiority designed comparative clinical trials, which led to their inclusion as first line options in all versions of HIV therapeutic guidelines.

Expert commentary: Some unprecedented clinical-pharmacological properties of INSTIs, such as their rapid and sustained action against HIV replication, the optimal tolerability and safety profile and a clinically proven robust genetic barrier are the main factors justifying the successful clinical use of INSTIs. Based on these unique features, novel INSTIs-based treatment modalities are being developed, including the reduction of antiretroviral regimens to two drugs only.     

HIV integrase inhibitors: a new era in the treatment of HIV

Introduction: Integrase inhibitors (INIs) are the latest class of antiretroviral drugs approved for the treatment of HIV infection and are becoming ‘standard’ drugs in the treatment of both naïve as well as heavily pretreated individuals with HIV.

Areas covered: Data on efficacy, safety, tolerability, pharmacokinetics, drug-drug interactions and resistance are reviewed from the pivotal Phase III clinical trials published in PubMed high-impact medical journals or presented at international meetings.

Expert opinion: Due to their outstanding data of efficacy, tolerability, safety – shared by all three drugs (raltegravir, elvitegravir, dolutegravir) currently belonging to this new family of antiretrovirals – INIs have become part of the recommended initial antiretroviral therapy options. Some differences in dosing, drug-drug interactions and robustness/genetic barrier among the three drugs will provide the physician the characteristics to make the best choice.     

3-Hydroxy-6,7-dihydropyrimido[2,1-c][1,4]oxazin-4(9H)-ones as new HIV-1 integrase inhibitors WO2011046873 A1

Since the discovery of raltegravir, the first FDA-approved integrase inhibitor, Merck and other pharmaceutical companies have continued their research programs in order to introduce novel molecules as second generation integrase inhibitors. Elvitegravir (Japan Tobacco/Gilead) and dolutegravir (Shionogi/GlaxoSmithKline) are in advanced stages of clinical development. Bristol-Myers Squibb has developed molecules leading to BMS-707035, which was stopped at the Phase II clinical trial stage. Herein is presented the last patent from this company where, in particular, new 3-hydroxy-6,7-dihydropyrimido[2,1-c][1,4]oxazin-4(9H)-ones are synthesized and their biological properties given.