Targeting host deubiquitinases for broad spectrum anti-infective therapy

迫切需要有效靶向耐药的新疗法。 微生物和病原体，目前没有治疗方案。诺如病毒引起 估计有2300万人感染， 但是没有抗病毒药物或疫苗可以治疗或预防感染。 单核细胞增生李斯特菌是另一种食源性致病菌，致死率高达20- 25%，是最近香瓜相关疫情的罪魁祸首。快速诊断a 在疾病爆发期间，特定病原体的检测往往具有挑战性。因此， 开发广谱治疗药物的基本原理， 传染病的治疗。实现这一目标的一种方法是发展“反- 感染性”化合物靶向宿主编码的蛋白质在感染过程中的关键 多种微生物我们发现一个小分子，WP 1130（WP）和相关的 化合物，表现出抗多种病原体的抗感染活性，包括 几种B类病原体：细菌（MRSA、单核细胞增生李斯特菌、沙门氏菌 肠血清型鼠伤寒），病毒（鼠和人诺如病毒， 脑心肌炎病毒、辛德毕斯病毒、拉克罗斯病毒）和顶复体 弓形虫（Toxoplasma gondii）我们的初步研究表明，WP抑制一个子集， 宿主去泛素化酶（DUBs）的作用导致泛素化蛋白在宿主细胞中的积累， 细胞，但不直接影响宿主外的病原体。泛素是一种真核生物 一种低分子量多肽，当 与靶蛋白共价连接。WP类似物的研究为研究 开发新的治疗方法，通过干扰 宿主和微生物之间的关键相互作用。泛素化和去泛素化 宿主靶标在许多不同的微生物感染中起关键作用。因此我们 假设WP处理选择性地抑制关键DUB， 微生物病原体，从而限制感染。我们将使用鼠诺如病毒和L。 单核细胞增多症作为两个不相关的病原体与建立的小动物模型， 感染，以确定介导WP抗感染作用的DUB靶点。我们的目标是 将先导化合物推向研究性新药（IND）申请。 因此，我们提出以下具体目标：（1）确定WP的目标DUBs， 介导抗感染活性，（2）体外测试SAR系列WP以鉴定铅 化合物，和（3）测试先导化合物的体内功效。本申请旨在 开发一种广谱抗感染治疗药物，对多种病原体有效 包括多种B类药剂。

There is an urgent need for new therapeutics that effectively target drug-resistant microbes and pathogens that currently have no treatment options. Noroviruses cause an estimated 23 million infections and over half of all food-borne gastroenteritis outbreaks in the US every year but no antivirals or vaccines exist to treat or prevent infections. Listeria monocytogenes is another food-borne pathogen with a high fatality rate of 20- 25%, responsible for the recent cantaloupe-associated outbreak. Rapid diagnosis of a specific pathogen during outbreaks is often challenging. Thus, there is a compelling rationale for the development of broad-spectrum therapeutics for early and effective treatment of infectious diseases. One approach towards that goal is to develop “anti- infective” compounds that target host-encoded proteins critical during infection of multiple microbes. We find that a small molecule, WP1130 (WP) and related compounds, exhibit anti-infective activity against multiple classes of pathogens, including several category B pathogens: bacteria (MRSA, Listeria monocytogenes, Salmonella enterica serovar Typhimurium), viruses (murine and human norovirus, encephalomyocarditis virus, Sindbis virus, La Crosse virus), and an apicomplexan parasite (Toxoplasma gondii). Our initial studies demonstrate that WP inhibits a subset of host deubiquitinases (DUBs) resulting in the accumulation of ubiquitinated proteins in cells, but does not directly affect pathogens outside the host. Ubiquitin is a eukaryotic low molecular weight polypeptide that acts as a post-translational regulatory switch when covalently linked to target proteins. Studies of WP analogs hold vital clues for the development of novel therapeutics that may effectively contain pathogens by interfering with key interactions between host and microbe. Ubiquitination and deubiquination of host targets play critical roles in many different microbial infections. We therefore hypothesize that WP treatment selectively inhibits key DUBs exploited by microbial pathogens, thereby limiting infection. We will use murine norovirus and L. monocytogenes as two unrelated pathogens with established small animal models of infection to define DUB targets that mediate the anti-infective effects of WP. Our goal is to advance the lead compound towards an investigational new drug (IND) application. We therefore propose the following specific aims: (1) Determine target DUBs of WP that mediate anti-infective activity, (2) Test a SAR series of WP in vitro to identify lead compounds, and (3) Test lead compounds for in vivo efficacy. This application aims to develop a broad-spectrum anti-infective therapeutic effective against many pathogens including multiple category B agents.