Therapeutics that target processivity complex proteins of pox and other viruses

描述(由申请人提供)：阻止天花(天花的病原体)感染的新的安全疗法对于快速应对生物恐怖主义至关重要。治疗技术可以在疫苗交付延迟时防止天花传播，并保护患有疫苗禁忌症的患者。聚合酶-加工性复合体是理想的治疗靶点，因为它们具有选择特定(加工性)和广谱(聚合酶)抑制剂的潜力。加工性因素将DMA聚合酶与DMA捆绑在一起，使酶能够合成长链。我们现在已经确定，痘苗病毒(W)的过程性DMA合成需要三种蛋白质，E9DMA聚合酶和A20，D4R过程性因子。由于这三种痘苗病毒蛋白与天花病毒的相应蛋白至少有97%的同源性，W疗法被预测为针对天花病毒的相同功能。我们最近使用我们的快速平板法完成了对来自小型化学库的52,000种化合物的高通量筛选(HTS)。我们确定了21种先导抑制剂，它们可以有效地阻断痘苗病毒DNA的合成，防止感染，而细胞毒性可以忽略不计。第二个HTS将增加额外的线索。我们的目标是通过体外选择性试验、使用SPR技术和分析治疗耐药病毒来确定三联体(E9/A20/D4)的哪种蛋白是每个铅抑制剂的靶标。其他新的类似物将通过在硅化合物采矿中获得。精选先导化合物的药物化学将产生具有卓越抗病毒效力和安全性的抑制剂。令人兴奋的是，我们还将利用基于晶体D4-A20结合的合理药物设计来识别痘病毒的高度特异性抑制剂。先导化合物将在疾控中心测试对天花病毒的抑制作用。我们将评估先导化合物保护小鼠免受痘病毒攻击的效果。我们还将继续定义广谱抑制剂，以防止感染其他医学上重要的病毒。我们的研究旨在产生优秀的治疗方法，在未来可以进入药代动力学测试阶段。我们的希望是开发一种独特的抑制剂，以预防天花和其他病毒疾病。相关性(见说明)：这些研究旨在发现如果天花被用作生物恐怖武器，将防止天花传播的治疗方法。其中某些疗法的广谱作用有望产生新的抗病毒药物，可用于预防当前具有医学意义的感染，并阻止其他重要的生物恐怖主义因素。

DESCRIPTION (provided by applicant): Novel and safe therapeutics that block infection by variola, the causative agent of smallpox, are essential for a rapid response to bioterrorism. Therapeutics can prevent smallpox spread when vaccine delivery is delayed and protect individuals with conditions for whom the vaccine is contraindicated. Polymerase- Processivity complexes are ideal therapeutic targets in that they have the potential to select for both Specific (processivity) and Broad-Spectrum (polymerase) inhibitors. Processivity factors tether DMA polymerase to DMA, enabling the enzyme to synthesize long strands. We have now established that processive DMA synthesis of vaccinia virus (W) requires three proteins, an E9 DMA polymerase and the A20, D4R processivity factors. Because these three vaccinia proteins are at least 97% homologous to the corresponding proteins of variola virus, W therapeutics are predicted to target the same function in variola. We recently completed High Throughput Screen (HTS) of 52,000 compounds from small chemical libraries using our Rapid Plate Assay. We identified 21 LEAD inhibitors that effectively block vaccinia virus DNA synthesis and prevent infection with negligible cell cytotoxicity. A second HTS will add additional LEADS. Our goals are to define which protein of the triad (E9/A20/D4) is targeted by each LEAD inhibitor using an in vitro selectivity assay, employing SPR technology and analyzing therapeutic resistant viruses. Additional new analogs will be obtained by in silico Compound Mining. Medicinal Chemistry of select LEAD compounds will generate inhibitors of superior anti-viral potency and safety. Excitingly, we will also employ Rational Drug Design based on the crystal D4-A20 association to identify highly specific inhibitors of poxviruses. LEAD compounds will be tested for inhibition of variola virus at the CDC. We will evaluate the efficacy of LEAD compounds to protect mice against challenge by poxviruses. We will also continue to define Broad-Spectrum inhibitors that prevent infection of other medically important viruses. Our studies are intended to produce excellent therapeutics that can, in the future, be taken to the pharmokinetic stages of testing. Our hope is to develop a unique class of inhibitors that protect against pox and other viral diseases. RELEVANCE (See instructions): These studies are aimed at discovering therapeutics that will prevent the spread of smallpox, if it becomes used as a bio-terror weapon. The Broad-Spectrum action of certain of these therapeutics will hopefully generate new anti-virals that can be used to prevent current infections that are of medical significance as well as block other important agents of bioterrorism.