Cross-Kingdom Vaccine Targeting Healthcare-Associated Priority Pathogens

该申请描述了针对多药耐药（MDR）的广谱疫苗的开发 微生物，主要与卫生保健相关感染（HAI）有关。我们的前提是， 预防抗生素耐药性是通过疫苗，而不是不断引进新的药物。 我们的开发计划是基于真菌白色念珠菌的两种细胞壁抗原：Als 3 p，一种多细胞壁抗原， 功能adhesin/invasin;和Hyr 1 p，其使C.白色念珠菌逃避吞噬细胞的杀伤。接种小鼠 与任一种抗原联合使用，可显著预防念珠菌属引起的播散性感染 和念珠菌性外阴阴道炎（VVC）。白色念珠菌。重要的是，用两种抗原协同接种 保护小鼠免受VVC。 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 Als 3 p和耐甲氧西林葡萄球菌之间的三维（3-D）结构和功能同源性 金黄色葡萄球菌（MRSA）表面粘附分子，包括凝集因子A。类似地，Hyr 1 p，共享3-D结构 与MRSA SraP同源，是血小板粘附素。Hyr 1 p还与 耐多药鲍曼不动杆菌和产碳青霉烯酶克雷伯菌血凝素/溶血素 肺炎链球菌（CPKP）。用Als 3 p的重组N末端（rAls 3 p-N）主动免疫导致>50%的 在葡萄球菌血症的其他致命小鼠模型中存活，并保护小鼠免受皮肤和皮肤结构 MRSA引起的感染类似地，靶向免疫球蛋白的主动或被动免疫（用单克隆抗体）也可以用于免疫治疗。 Hyr 1 p的重组N-末端保护小鼠免受MDR AB和CPKP感染。因此，我们的愿景是发展 靶向念珠菌、MRSA和MDR AB和CPKP的“跨王国”双抗原疫苗。都是重要的领导 因为HAIs。具体目的是：a）通过协同作用评价优化双重rAlsp 3-N/rHyr 1 p-N疫苗， 抗原剂量的调整和临床安全的新佐剂的使用; B）进行GLP-使能研究，包括分析 测定开发/优化，优化的双抗原疫苗的制剂规模扩大和稳定性研究;和c） 评价含和不含抗生素的最终疫苗制剂的活性，并进行IND使能GLP毒性 研究优化的最终疫苗制剂。 与NovaDigm Therapeutics合作，我们推进了rAls 3 p-N的开发， 氢氧化铝（即NDV-3A）从学术实验室到1b/2a期试验，显示了 疫苗可保护40岁以下女性免于复发VVC。因此，该提案将利用我们的联合 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.