Discovery of antivirals against vaccinia and smallpox

发现针对痘苗和天花的抗病毒药物

• 批准号: 6562005
• 负责人: ROBERT Paul RICCIARDI
• 金额: $ 23.78万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6562005
• 关键词: DNA binding protein; DNA directed DNA polymerase; Poxviridae disease; antiviral agents; biotechnology; bioterrorism /chemical warfare; drug discovery /isolation; high throughput technology; nucleic acid inhibitor; proteomics; smallpox virus; vaccinia virus

DESCRIPTION (provided by applicant): There is a mounting concern that terrorist groups may possess or can obtain their own stocks of variola virus, the etiological agent of smallpox infection. To respond to this specific threat, the United States must have at its disposal supplies of both vaccinia virus vaccine and antiviral compounds directed against smallpox infection. The antiviral compounds are needed in situations in which the vaccine is contraindicated, such as immunosuppression. In addition, the antiviral compounds are needed to blunt adverse complications that are known to be associated with vaccine administration. An ideal antiviral should also be able to directly prevent smallpox infection in situations where vaccine delivery is delayed. Toward this end, antivirals directed against the vaccinia virus DNA polymerase (V AC-POL) and processivity factor (V AC-PF) have great potential. In general, PFs act as clamps that slide along the DNA and bind to their respective POLs. Through this POL/PF interaction, DNA synthesis will be processive, generating new DNA strands that are thousands of nucleotides long. Viruses that have mutated PFs are defective in replication since POL alone can synthesize only short DNA products (as few as three nucleotides). A strongly favorable feature of PFs is that they function with their own POLs, so that an antiviral that targets a viral PF should be very specific and not interfere with cellular replication. V AC-POL and V AC-PF interaction has been recently characterized and both proteins can be purified and combined to function in processive DNA synthesis. The goal of this project is to discover antivirals that will specifically block processive DNA synthesis by V AC-POL and V AC-PF. A newly developed mechanistic rapid plate assay will be used to screen thousands of potentially inhibitory compounds. This high throughput screening will be followed by procedures to evaluate the inhibitory compounds and to ultimately test their ability to block vaccinia virus infection. A drug that prevents vaccinia processive DNA synthesis will be useful in curtailing vaccinia vaccine complications. The same drug may also directly block variola infection since the POLs and PFs, respectively, of vaccinia and variola have 97% direct homology. The approach may help deliver the 'just-in-time' need for reagents to combat a smallpox bioterrorism threat.

描述（由申请人提供）：持续关注的是，恐怖组织可能拥有或可以获得自己的Variola病毒的股票，这是天花感染的病因学药。为了应对这一特定威胁，美国必须拥有针对天花感染的疫苗病毒疫苗和抗病毒化合物的供应。在禁忌疫苗（例如免疫抑制）的情况下，需要抗病毒化合物。另外，需要抗病毒化合物来钝化与疫苗给药相关的不良并发症。理想的抗病毒也应能够直接防止在疫苗输送延迟的情况下直接防止天花感染。为此，针对疫苗病毒DNA聚合酶（V AC-POL）和加工性因子（V AC-PF）的抗病毒药具有很大的潜力。通常，PFS充当沿DNA滑动并与它们各自的pol结合的夹具。通过这种POL/PF相互作用，DNA合成将是过程的，产生了数千个核苷酸长的新DNA链。突变PFS的病毒在复制中有缺陷，因为仅POL可以合成短DNA产物（只有三个核苷酸）。 PFS的一个非常有利的特征是它们可以与自己的pol起作用，因此靶向病毒PF的抗病毒药应非常具体，并且不会干扰细胞复制。 V AC-POL和V AC-PF相互作用最近已经表征，并且可以将两种蛋白质纯化并组合起来，以在过程中的DNA合成中起作用。该项目的目的是发现抗病毒药，该抗病毒药将特异性阻断VAC-POL和V AC-PF的过程DNA合成。新开发的机械快速板测定法将用于筛选数千种潜在的抑制性化合物。这种高吞吐量筛查将进行评估抑制性化合物的程序，并最终测试其阻断疫苗病毒感染的能力。一种防止疫苗的加工DNA合成的药物将有助于减少疫苗疫苗并发症。同一药物也可能直接阻止Variola感染，因为疫苗和Variola的POL和PFS分别具有直接同源性的97％。这种方法可能有助于提供对试剂的“及时”需求，以应对天花生物恐怖主义威胁。