Therapeutics that target processivity complex proteins of pox and other viruses

描述（由申请人提供）：阻断天花病原体天花感染的新型安全疗法对于快速应对生物恐怖主义至关重要。治疗方法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个体。聚合酶-聚合性复合物是理想的治疗靶点，因为它们具有选择特异性（聚合性）和广谱（聚合酶）抑制剂的潜力。加工因子将DMA聚合酶拴在DMA上，使该酶能够合成长链。我们现在已经确定，牛痘病毒(W)的程序性DMA合成需要三种蛋白质，E9 DMA聚合酶和A20， D4R加工因子。由于这三种牛痘蛋白与天花病毒的相应蛋白至少有97%的同源性，预计W疗法将针对天花病毒的相同功能。我们最近完成了高通量筛选（HTS） 52000个化合物从小型化学文库使用我们的快速板分析。我们鉴定出21种有效阻断牛痘病毒DNA合成并以可忽略的细胞毒性预防感染的铅抑制剂。第二个HTS将增加额外的引线。我们的目标是通过体外选择性试验，采用SPR技术和分析治疗性耐药病毒，确定每种LEAD抑制剂针对三联体（E9/A20/D4）中的哪个蛋白。通过硅化合物开采将获得更多新的类似物。通过对铅化合物的药物化学研究，可以产生具有较强抗病毒效力和安全性的抑制剂。令人兴奋的是，我们还将采用基于晶体D4-A20关联的Rational Drug Design来识别高度特异性的痘病毒抑制剂。铅化合物将在疾病预防控制中心测试对天花病毒的抑制作用。我们将评估铅化合物保护小鼠免受痘病毒攻击的功效。我们还将继续确定广谱抑制剂，以防止其他医学上重要的病毒感染。我们的研究旨在生产出优秀的治疗方法，以便在未来进入药物动力学测试阶段。我们的希望是开发一种独特的抑制剂，防止痘和其他病毒性疾病。相关性（见说明）：这些研究旨在发现治疗方法，在天花被用作生物恐怖武器时防止其传播。这些治疗方法中的某些广谱作用有望产生新的抗病毒药物，可用于预防具有医学意义的当前感染，并阻止其他重要的生物恐怖主义制剂。

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.