Design of Antiviral Drugs Effective Against the HIV-1 Vi

有效对抗 HIV-1 Vi 的抗病毒药物的设计

• 批准号: 6762126
• 负责人: ETTORE APPELLA
• 金额: --
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6762126
• 关键词: AIDS; AIDS therapy; HIV infections; antiAIDS agent; antiviral agents; chemical structure function; drug design /synthesis /production; drug screening /evaluation; epitope mapping; human immunodeficiency virus 1; nucleocapsid; phenylamide; protein sequence; protein structure function; sulfides; tissue /cell culture; virion; virus RNA; virus antigen

In earlier work, we have described a novel chemotype, the pyridinioalkanoyl thioesters (PATEs) as anti-HIV-1 agents that selectively target the viral nucleocapsid protein zinc fingers without affecting other viral or host zinc fingers. Our efforts during the last several years have been devoted to finding compounds with substantially improved in vitro antiviral potencies. An entirely different class of ligand structures, based on simple aliphatic amino acid amides, was uncovered (Song et al. Bioorg Med Chem.10:1263-73, 2002; Goel et al. Bioorg Med Chem Lett.12:767-70, 2002). This past year, we have synthesized suitable S-acyl 2-mercaptobenzamide derivatives that are poor substrates for non-specific carboxyesterases (thioesterases) yet retain good antiviral activity. We have explored this possibility by introducing a variety of substituents on the acyl group and the benzamide head structure that modify the reactivity of the thioester bond through electronic influences and provide steric hindrance to the approach of enzymes. So far, we have prepared about 50 representatives of this chemotype and evaluated them for anti-HIV activity via the dose-response inhibition of p24 secreted in vitro by activated spleen cells taken from HIV-1 transgenic mice and via the in vitro XTT cytoprotection assay. All the compounds showed EC50 values between 1.9 and 31.8 mM, and exhibited a wide range of cellular toxicity (IC50) values between 22 and 790 mM in the XTT assay. EC50s between 2.2 and 15.2 mM and IC50s of 59 and >250 mM were obtained in the Tg assay. Introduction of substituents in the phenyl ring of the acyl group modified the reactivity of the thioester bond through an electronic effect and also provided steric hindrance that reduced hydrolysis of the thioester bond. The presence of an electron donating substituent such as OCH3 showed enhanced antiviral activity but the half-life was only 80 mins. On the other hand, the presence of the same substituent at a different position or at more than one position in the phenyl ring, allowed greater stability but decreased antiviral activity. Compounds where the substituents are bulky in nature, for example Cl or CH3 at position 2 and 3 of the phenyl ring, showed low EC50s and high toxicity. Moreover, a compound having a tertiary butyl moiety at the acyl group was highly stable with a half-life of 335 minutes, low EC50 value and moderate cytotoxicity. Thus we have accomplished one of our goals for this year through the identification of a new chemotype with promising potential for development as an HIV drug. Recently, some of the lead compounds were further evaluated for antiviral efficacy in combination with nucleoside reverse transcriptase inhibitors (NRTI; AZT, PMPA), nonnucleoside reverse transcriptase inhibitors (NNRTI; Efavirenz [EFV], Nevaripine [NVP]), Protease inhibitors (PI; Rotinavir [RTV], Indinavir [IDV]), Zn finger inhibitors (ZFI; azodicarbonamide [ADA]) and virus entry inhibitors (EI; Chicago Sky Blue [CSB] and T-20) using the Prichard and Shipman MacSynergy 2 proportional statistics model (Prichard and Shipman (1990) Antiviral Res. 14:181-206). Our results show that the PATE chemotype via targeting retroviral Zn fingers can interact synergistically with all classes of clinically relevant antivirals and that the Zn fingers inhibitors could potentially play an important role in antiviral therapy by providing a fourth class of antivirals for treatment. The discovery of this family of highly potent NCp7 inhibitors holds promise for inactivating all strains of HIV-1 without generating resistant strains.

在较早的工作中，我们描述了一种新型的化学型，吡啶烷烷酰基硫烷硫代酯（PATES）是抗HIV-1药物，它们在不影响其他病毒或宿主锌指的情况下选择性地靶向病毒核素蛋白锌手指。在过去的几年中，我们的努力一直致力于寻找具有显着改善体外抗病毒药效力的化合物。发现了完全不同类别的配体结构，基于简单的脂肪族氨基酸酰胺，发现了（Song etal。BioorgMed Chem.10：1263-73，2002; Goel等人Bioorg Med Chem Lett.12：767-70，2002）。在过去的一年中，我们合成了合适的S-acyl 2-甲苯二苯衍生物，这些衍生物是非特异性羧酸酯酶（硫酯酶）的底物，但保留了良好的抗病毒活性。我们通过在酰基基团和苯甲酰乳头结构上引入各种取代基来探讨这种可能性，从而通过电子影响来改变硫酯键的反应性，并为酶方法提供空间的阻碍。到目前为止，我们已经准备了该化学型的大约50个代表，并通过通过从HIV-1转基因小鼠和体外XTT细胞保护分析的体外激活的脾脏细胞对P24的剂量反应抑制对它们进行了抗HIV活性。所有化合物均显示1.9至31.8 mm之间的EC50值，并在XTT分析中显示了22至790 mm之间的各种细胞毒性（IC50）值。在TG分析中获得了2.2至15.2 mm和IC50的EC50，IC50为59和> 250 mm。在酰基基团的苯基环中引入取代基修饰了硫酯键的反应性，并提供了降低硫酯键水解的空间障碍。捐赠取代基（例如OCH3）的存在表现出增强的抗病毒活性，但半衰期仅为80分钟。另一方面，在不同位置或在苯环中多个位置处的相同取代基的存在使得稳定性更高，但抗病毒活性降低。化合物的化合物本质本质上是笨重的，例如苯环第2和3位的Cl或CH3显示出低EC50和高毒性。此外，在酰基基团的三级丁基部分的化合物高度稳定，半衰期为335分钟，EC50值低和中等的细胞毒性。因此，通过鉴定新的化学型，我们已经实现了今年的目标之一，并具有有希望的发育潜力作为艾滋病毒药物。 最近，进一步评估了某些铅化合物的抗病毒疗效，以与核苷逆转录酶抑制剂（NRTI； AZT，PMPA），非核苷逆转录酶抑制剂（NNRTI; efavirenz; efavirenz [efv]，nvaripine [nvp]，prot inir Inir infir inir inf; [RTV]，Indinavir [IDV]），使用Prichard和Shipman MacSynergy 2比例统计模型（Prichard and Flagman and Flagman and Flagman and Flagman and Flagman（1990年），使用Prichard和Shipman MacSynergy 2 res。我们的结果表明，通过靶向逆转录病毒Zn手指的PATE化学型可以与所有类别相关的临床相关抗病毒药协同相互作用，并且Zn手指抑制剂可以通过为抗病毒药疗法发挥重要作用，通过为抗病毒药疗法发挥重要作用。这个高度有效的NCP7抑制剂家族的发现有望在不产生抗性菌株的情况下灭活所有HIV-1菌株。