Novel selective substituted aminofurazans kinase inhibitors for Drug resistant Bacteria

抗生素耐药性细菌是对人类健康的主要威胁。每年在美国， 大约200万人感染了对抗生素具有抗药性的细菌，导致至少 23000人死亡。开发新的抗生素是必要的。然而，从头开始抗生素的开发 资源密集型。鉴于投资回报不佳，各大制药公司大幅 减少了抗生素的研发工作。学术实验室通常无法访问 化合物和足够的数据来进行合理的设计。因创伤接受外科手术的退伍军人， 包括头部创伤在内，容易受到多种外科感染，有时是不太可能发生的细菌感染 对制药公司来说，它永远是一个有吸引力的目标。 利用工业进行的现有药物设计工作进行抗生素开发可能会避免 一些由于不良的药理特性而造成的代价高昂的失误可能会不经意地出现在 选择。我们的方法使用基于结构的计算模型来将药物重新用作抗生素。 我们实验室以前的工作表明，抑制属于青霉素家族的一种李斯特菌激酶 结合蛋白和丝氨酸/苏氨酸激酶相关蛋白可使细菌对β-内酰胺敏感 抗生素。细菌丝氨酸/苏氨酸激酶是一种信号转导分子，我们的工作表明它们是一种新的 药物开发目标。虽然一些革兰氏阳性细菌中常见的是意大利面激酶，但我们发现 药物选择性，不仅在人类和细菌激酶之间，而且在细菌激酶之间也是如此。我们是 用一个大的后袋瞄准含有面条激酶的细菌的子集。我们已经得到了 我们的目标细菌激酶(PounB)的晶体结构，其中一个铅化合物证明了这一点。这 通过使用信息，我们可以专门修改销售线索以提高效力并最大限度地提高选择性 先前定义的与人类激酶的结构活性关系。此外，我们的原子分辨药物： 靶结构将指导我们对药物的区域进行修改，以明确测试利平斯基的规则 药物开发，并确定是否应该为了抗生素药物开发而对它们进行修改。在这份提案中， 我们将在现有工作的基础上再接再厉，实现以下具体目标： 目标1：设计、合成和测试生化和微生物方面的新型氨基呋喃 利用一些蛋白酪氨酸酶家族的独特形状的后袋的蛋白激酶抑制剂。 目的2：测试这些新型激酶抑制剂的毒性，并确定其有效性。 在动物模型(Danio Rerio)中将β-内酰胺类抗生素与激酶抑制相结合。我们会 分析我们的激酶抑制剂对一组人类激酶以及对 斑马鱼的胚胎发生和繁殖力。 当这项工作完成时，我们将产生关于如何设计作用不佳的激酶抑制剂的数据 对人类和斑马鱼的酶有抗药性，但对某些细菌的酶有很强的抗药性。病原体包括 具有这种独特的后袋形态的激酶包括各种皮肤和土壤细菌，使其复杂化 退伍军人中的创伤性和外科感染，如梭状芽胞杆菌、诺卡氏菌、痤疮丙酸杆菌和 分枝杆菌。此外，我们的工作将为治疗耐药性的早期试验奠定基础 分枝杆菌和诺卡氏菌感染以及可能使外科手术复杂化的非典型皮肤微生物 退伍军人中常见的感染，使用一种新的药物，该药物与现有的低浓度抗生素类有协同作用 毒性。

Antibiotic resistant bacteria are a major threat to human health. Each year in the United States, approximately 2 million people are infected with bacteria that are resistant to antibiotics resulting in at least 23,000 deaths. The development of novel antibiotics is needed. However, de novo antibiotic development is resource intensive. Given the poor return on investment, major pharmaceutical companies have significantly decreased antibiotic Research and Development efforts. Academic labs generally do not have access to compounds and sufficient data for rationale design. Veterans who undergo surgical procedures for trauma, including head trauma, are vulnerable to multiple surgical infection, sometimes with organisms that are unlikely to ever represent an attractive target for pharmaceutical companies. Antibiotic development that capitalizes on existing drug designwork conducted by industry, may avoid some of the expensive missteps due to poor pharmacologic properties that can inadvertently arise in the lead selection. Our approach uses structure-based computational modeling to repurpose drugs as antibiotics. Previous work in our lab demonstrated that inhibition of a Listeria kinase, belonging to a family of Penicillin binding And Serine/Threonine kinase Associated (PASTA) proteins, can sensitize the bacteria to beta-lactam antibiotics. Bacterial Serine/Threonine kinases are signal transducers and our work suggests they are a novel drug development target. While PASTA kinases are common in some gram-positive bacteria, we found there is drug selectivity, not only between human and bacterial kinases, but also between bacterial kinases. We are targeting the subset of PASTA kinase containing bacteria with a large back pocket. We have already obtained a crystal structure of our target bacterial kinase (PknB) with a lead compound demonstrating this pocket. This information enables us to specifically modify the lead to both increase potency and maximize selectivity using previously defined structure activity relationships with human kinases. Furthermore, our atomic resolution drug: target structure will guide us on regions of the drug that can be modified to explicitly test Lipinski's rules for drug development and determine if they should be modified for antibiotic drug development. In this proposal, we will build on our existing work with the following Specific Aims: Aim 1: Design, synthesize, and test both biochemically and microbiologically novel aminofurazan kinase inhibitors that exploit the uniquely shaped back pocket of some of the PknB family of kinases. Aim 2: Test the toxicity of these novel kinase inhibitors, as well as determine the effectiveness of combined beta lactam antibiotics with kinase inhibition in an animal model (Danio rerio). We will analyze the effect of our kinase inhibitors on both a panel of human kinases as well as on embryogenesis and fertility in zebrafish. When this work is complete, we will have generated data on how to design kinase inhibitors that act poorly against human and zebrafish kinases, but are potent against certain bacterial kinases. Pathogens that include kinases with this unique back pocket morphology include diverse skin and soil bacteria that complicate traumatic and surgical infections in the VA such as Clostridia, Nocardia, Propionibacterium acnes, and Mycobacterium. Furthermore, our work will set the stage for early stage trials on treating drug resistant mycobacterial and nocardial infections as well as atypical skin organisms that can complicate surgical infections common in veterans using a novel agent that synergizes with an existing antibiotic class with low toxicity.