Cross-Kingdom Vaccine Targeting Healthcare-Associated Priority Pathogens

本申请描述了针对多药耐药(MDR)的广谱疫苗的开发 生物体，主要与医疗保健相关感染(HAI)有关。我们的前提是，有效的方法是 预防抗菌素耐药性是通过疫苗而不是继续推出新药来实现的。 我们的开发计划是基于白色念珠菌的两种细胞壁抗原：Als3p，一种多菌体 功能粘附素/粘附素；以及Hyr1p，使白色念珠菌能够逃避吞噬细胞的杀伤。为小鼠接种疫苗 任何一种抗原对念珠菌引起的播散性感染具有显著保护作用 以及由白色念珠菌引起的外阴阴道念珠菌病(VVC)。重要的是，两种抗原的疫苗接种具有协同作用 保护小鼠免受VVC的侵袭。 使用创新的计算分子建模和生物信息学策略，我们确定了三个非常重要的 Als3p与耐甲氧西林葡萄球菌的三维结构和功能同源性 金黄色葡萄球菌(MRSA)表面黏附分子，包括聚集因子A。类似地，Hyr1p具有三维结构 与MRSA SRAP同源，一种血小板粘附素。Hyr1p在结构上也与 耐多药鲍曼不动杆菌和产碳青霉烯酶克雷伯菌的血凝素/溶血素 肺炎(CPKP)。用重组Als3p(rAls3p-N)的N末端主动免疫可导致50% 在致命性葡萄球菌血症小鼠模型中存活并保护小鼠免受皮肤和皮肤结构的影响 金黄色葡萄球菌感染。同样地，针对病毒的主动或被动免疫(使用单抗) 重组Hyr1p N末端对MDR AB和CPKP感染小鼠的保护作用因此，我们的愿景是发展 一种针对念珠菌、耐甲氧西林金黄色葡萄球菌、多药耐药AB和CPKP的“跨王国”双抗原疫苗。所有这些都是重要的领导 HAIS的起因。具体目标是：a)通过协同评价优化rAlsp3-N/rHyr1p-N双联疫苗，精细化 调整抗原剂量，并使用临床上安全的新型佐剂；b)进行支持GLP的研究，包括分析 测试开发/优化、优化双抗原疫苗的配方放大和稳定性研究；以及c) 评估最终疫苗配方在使用和不使用抗生素的情况下的活性，并进行启用IND的GLP毒性 优化最终疫苗配方的研究。 在与NovaDigm Treeutics的合作下，我们推动了rAls3p-N的开发 氢氧化铝(即新城疫病毒3A)从学术实验室转移到1b/2a阶段试验，显示了 疫苗在预防40岁妇女VVC复发中的作用。因此，这项提案将利用我们的联合 融合疫苗抗原在基础发现和产品开发方面的优势 人类病原体。

This application describes development of a broad-spectrum vaccine targeting multidrug resistant (MDR) organisms, principally related to healthcare-associated infections (HAIs). Our premise is that an effective way to prevent antimicrobial resistance is through vaccines rather than continued introduction of new drugs. Our development program is based on two cell wall antigens of the fungus Candida albicans: Als3p, a multi- function adhesin/invasin; and Hyr1p, which enables C. albicans to evade phagocyte killing. Vaccination of mice with either antigen provides significant protection against disseminated infections caused by Candida species and vulvovaginal candidiasis (VVC) due to C. albicans. Importantly, vaccination with both antigens synergistically protect mice from VVC. Using innovative computational molecular modeling and bioinformatic strategies, we identified highly significant three dimensional (3-D) structural and functional homology between Als3p, and methicillin resistant Staphylococcus aureus (MRSA) surface adhesive molecules, including clumping factor A. Similarly, Hyr1p, shares 3-D structural homology to MRSA SraP, an adhesin to platelets. Hyr1p also shares striking structural homology with hemagglutinin/hemolysin of MDR Acinetobacter baumannii (AB) and carbapenemase-producing Klebsiella pneumoniae (CPKP). Active immunization with the recombinant N terminus of Als3p (rAls3p-N) results in >50% survival in an otherwise fatal murine model of staphylococcemia and protects mice from Skin and Skin Structure Infection due to MRSA. Similarly, active or passive immunization (with a monoclonal antibody) targeting the recombinant N-terminus of Hyr1p protects mice from MDR AB, and CPKP infections. Thus, our vision is to develop a “cross-kingdom” dual antigen vaccine targeting Candida, MRSA, and MDR AB and CPKP. All are important leading cause of HAIs. Specific aims are: a) optimization of the dual rAlsp3-N/rHyr1p-N vaccine by synergy evaluation, fine- tuning of antigen dose, and use of clinically-safe newer adjuvants; b) conduct GLP-enabling studies including analytical assay development/optimization, formulation scale up of an optimized dual antigen vaccine and stability studies; and c) evaluate the final vaccine formulation for activity with and without antibiotics and perform an IND-enabling GLP-toxicity study of the optimized final vaccine formulation. In collaboration with NovaDigm Therapeutics, we advanced the development of rAls3p-N formulated with aluminum hydroxide (i.e. NDV-3A) from the academic laboratory to a phase 1b/2a trial showing efficacy of the vaccine in protecting women <40 years old from recurrent VVC. Thus, this proposal will leverage our combined strengths in basic discovery and product development of convergent vaccine antigens protecting against diverse human pathogens.