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.

在早期的工作中，我们已经描述了一种新的化学型，吡啶烷酰基硫酯（PATE）作为抗HIV-1药物，选择性靶向病毒核衣壳蛋白锌指，而不影响其他病毒或宿主锌指。在过去的几年中，我们的努力一直致力于寻找具有显著改善的体外抗病毒效力的化合物。发现了基于简单脂肪族氨基酸酰胺的完全不同种类的配体结构（Song等，Bioorg Med Chem.10：1263-73，2002; Goel等，Bioorg Med Chem Lett.12：767-70，2002）。在过去的一年中，我们已经合成了合适的S-酰基2-巯基苯甲酰胺衍生物，它们是非特异性羧酸酯酶（硫酯酶）的不良底物，但保留了良好的抗病毒活性。我们已经探索了这种可能性，通过引入各种取代基上的酰基和苯甲酰胺头部结构，通过电子的影响，修改的硫酯键的反应性，并提供空间位阻的酶的方法。到目前为止，我们已经准备了约50个代表这种化学型，并评估他们的抗艾滋病毒活性通过剂量反应抑制p24分泌体外活化脾细胞从HIV-1转基因小鼠，并通过在体外XTT细胞保护试验。在XTT试验中，所有化合物均显示出1.9至31.8 mM的EC 50值，并显示出22至790 mM的宽范围细胞毒性（IC 50）值。在Tg测定中获得2.2和15.2 mM之间的EC 50和59和>250 mM的IC 50。在酰基的苯环中引入取代基通过电子效应修饰了硫酯键的反应性，并且还提供了减少硫酯键水解的空间位阻。给电子取代基如OCH 3的存在显示出增强的抗病毒活性，但半衰期仅为80分钟。另一方面，在苯环的不同位置或多个位置存在相同的取代基，可以提高稳定性，但会降低抗病毒活性。其中取代基在性质上是大体积的化合物，例如苯环的2位和3位上的Cl或CH 3，显示出低EC 50和高毒性。此外，在酰基上具有叔丁基部分的化合物高度稳定，半衰期为335分钟，EC 50值低，细胞毒性中等。因此，我们通过鉴定一种新的化学型，实现了我们今年的目标之一，这种化学型具有开发艾滋病毒药物的良好潜力。 最近，一些先导化合物与核苷类逆转录酶抑制剂联合应用的抗病毒效果得到了进一步的评价（NRTI; AZT，PMPA），非核苷类逆转录酶抑制剂（NNRTI;依法韦仑[EFV]，奈瓦立平[NVP]），蛋白酶抑制剂（PI;罗替那韦[RTV]，茚地那韦[IDV]），锌指抑制剂（ZFI;偶氮二甲酰胺[ADA]）和病毒进入抑制剂（EI;芝加哥天蓝[CSB]和T-20），使用Prichard和Shipman MacSynergy 2比例统计模型（Prichard和Shipman（1990）Antiviral Res.14：181-206）。我们的研究结果表明，通过靶向逆转录病毒锌指的PATE化学型可以与所有类别的临床相关的抗病毒药物协同作用，并且锌指抑制剂可能通过提供第四类抗病毒药物用于治疗而在抗病毒治疗中发挥重要作用。这个高效NCp 7抑制剂家族的发现有望灭活所有HIV-1菌株，而不会产生耐药菌株。