Antivirals directed to novel targets in the virus-coded proteinase of adenovirus

用于治疗病毒感染的武器相对匮乏。虽然疫苗是为了 有些病毒可能是有效的，对于其他病毒，需要抗病毒药物。在潜力中 在某些病毒感染期间出现的抗病毒治疗的靶点是病毒编码的蛋白酶。 这些对合成传染性病毒必不可少的酶，是处理病毒特异性所必需的。 与这种病原体人类成熟、组装和复制有关的前体蛋白 病毒就像腺病毒一样。脊髓灰质炎病毒。丙型肝炎病毒和人类免疫缺陷病毒。病毒编码 蛋白酶对其底物具有高度的特异性，因此如果同样特定的抑制剂可以 它们应该干扰病毒的复制，而不是正常的细胞新陈代谢。我们的 开发新型抗病毒药物的模式系统是人腺病毒。一个明确的目标 就是在生化和结构水平上了解腺病毒的活性 蛋白水解酶(AVP)受调控。我们的实验室显示AVP是不活跃的，需要 在空间和时间上限制其活动的辅助因素。其中一个辅因子是pVIc，它是一种11个氨基酸 另一种是病毒DNA；肌动蛋白是一种细胞辅因子。辅因增加了kcat/km 用于底物水解。我们测定了AVP-pVIc的晶体结构，分辨率为1.6A 和AVP到0.98A。蛋白质的折叠是唯一的；AVP代表 新一类半胱氨酸蛋白酶。它通过54个氨基酸的长信号被pVIc激活 转导途径。另一个具体目标是使用这些信息来识别药物靶标 副总经理。在它的辅因中。并利用基于结构的药物设计来发现 与这些靶标结合的化合物。这项工作的一个新方面是识别药物靶标。 而不是活动站点中的那些。由于信号转导途径，超过25%的 AVP的表面是一个合法的药物靶点。有一种化合物，一种途径阻滞剂，据预测 从活性部位结合约15埃；它抑制pVIc激活AVP。一类新奇的 抑制剂是设计的-底物抑制剂。它们由此结合到底物上的裂解位点 阻止其被蛋白酶识别。尽管这项拨款提案的重点是获得 抗腺病毒药物，我们描述了从这个项目中产生的想法如何 应用于抗SARS冠状病毒和流感病毒的抗病毒药物的开发。

The arsenal of weapons for treating virus infections is relatively meager. Although vaccines for some viruses can be effective, for other viruses, antiviral agents are needed. Among potential targets for antiviral therapy that arise during certain viral infections are the virus-coded proteinases. These enzymes, essential for the synthesis of infectious virus, are required to process virus-specific precursor proteins involved in the maturation, assembly and replication of such pathogenic human viruses as adenovirus. poliovirus. hepatitis C virus, and human immunodeficiency virus. Viruscoded proteinases are highly specific for their substrates such that if equally specific inhibitors can be obtained, they should interfere with virus replication and not normal cellular metabolism. Our model system for the development of new antiviral agents is human adenovirus. One specific aim is to understand at the biochemical and structural levels how the activity of the adenovirus proteinase (AVP) is regulated. Our laboratory has shown that AVP is inactive and requires cofactors that restrict its activity in both space and time. One cofactor is pVIc, an 11 amino acid viral peptide; another is the viral DMA; actin is a cellular cofactor. The cofactors increase the kcat/Km for substrate hydrolysis. We determined the crystal structure of AVP-pVIc to a resolution of 1.6 A and of AVP to 0.98 A. The fold of the protein was unique; AVP represents the first member of a new class of cysteine proteinases. It is activated by pVIc via a 54 amino acid long signal transduction pathway. The other specific aim is to use this information to identify drug targets in AVP. in its cofactors. and in its substrates and to use structure-based drug design to discover compounds that bind to these targets. A novel aspect of the work is in identifying drug targets other than those in the active site. Because of the signal transduction pathway, more than 25% of the surface of AVP is a legitimate drug target. One compound, a pathway blocker, is predicted to bind about 15 Angstroms from the active site; it inhibits activation of AVP by pVIc. A novel class of inhibitors was designed- substrate inhibitors. These bind to a cleavage site on a substrate thereby preventing its recognition by the proteinase. Although the focus of this grant proposal is to obtain antiviral agents against adenovirus, we describe how ideas that arose out of this project can be applied to the development of antiviral agents against SARS Coronavirus and influenza virus.