Pn3Pase as an Effective Enzyme Therapeutic for Streptococcus Pneumoniae type 3 Infection

Pn3Pase 作为治疗 3 型肺炎链球菌感染的有效酶疗法

• 批准号: 10324274
• 负责人: Fikri Y Avci
• 金额: $ 29.95万
• 依托单位: PNEUMOTACTIX, LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-09 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10324274
• 关键词: Acetylation; Aftercare; Animals; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Autoimmune Responses; Bacteria; Bacterial Infections; Biological Assay; Biological Sciences; Blood Circulation; Boston; COVID-19; Cell-Mediated Cytolysis; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; Child; Clinical; Clinical Trials; Communicable Diseases; Conjugate Vaccines; Data; Death Rate; Defense Mechanisms; Detection; Disease; Dose; Drug Kinetics; Drug resistance; Effectiveness; Elderly; Enzymes; Epidemiologist; Fermentation; Genes; Genetic Recombination; Goals; HL-60 Cells; Half-Life; Hydrolysis; Immune; Immune response; Immunocompromised Host; Immunodeficient Mouse; In Vitro; Incidence; Individual; Infection; Infectious Agent; Influenza; Lead; Legal patent; Length; Mammalian Cell; Mediating; Modeling; Modification; Monosaccharides; Morbidity - disease rate; Mus; Patients; Phagocytes; Phagocytosis; Pharmacotherapy; Pneumococcal Infections; Pneumococcal vaccine; Polysaccharides; Population; Prevention; Primates; Proteins; Publishing; Resistance; Role; Sampling; Sepsis; Serotyping; Severities; Streptococcus pneumoniae; System; Testing; Therapeutic; Therapeutic Agents; Therapeutic Uses; Time; Toxic effect; Universities; Vaccination; Vaccine Research; Vaccines; Virulence; Virulent; Virus Diseases; bacterial resistance; base; capsule; commercialization; cytotoxicity; dosage; drug resistant pathogen; epidemiology study; experimental study; immune clearance; immunogenic; immunogenicity; in vivo; in vivo Model; intraperitoneal; macrophage; mortality; novel; novel therapeutics; pandemic disease; pathogen; prevent; prophylactic; research and development; secondary infection; therapeutic enzyme; therapy design; vaccine acceptance

Project Summary Streptococcus pneumoniae (Spn) remains one of the deadliest infectious agents responsible for more than a million deaths worldwide per year as well as making up a majority of secondary infections in immunocompromised patients. One of the major virulence mechanisms associated with this pathogen is its polysaccharide capsule, which determines successful colonization and infection of the bacterium as well as serving as a defense mechanism against immune-mediated killing of the pathogen. Spn serotype 3 (Spn3) is considered one of the most virulent serotypes of Spn, and despite current multivalent vaccines that include conjugate targets against Spn3, the overall proportion of this serotype in Spn infection cases has actually increased over the last ten years. Furthermore, Spn3 is directly associated with increased antibiotic resistance. Thus, the severity of the impact of Spn3, as well as the difficulties arising in designing therapies against it, cannot be overstated. Pneumozyme (Pn3Pase) is an enzyme product that we have previously shown to be highly effective in degrading the capsular polysaccharide of type 3 Spn. Our in vitro studies have shown that treating Spn3 with Pneumozyme will render the bacterium more susceptible to killing mechanisms by phagocytic cells. Furthermore, mice that are lethally challenged with Spn3 can be rescued following treatment with Pneumozyme: 100% of mice receiving Pneumozyme treatment survival the challenge whereas 100% of mice that receive inactive enzyme or vehicle die within two days of Spn3 lethal challenge. These results indicate that Pneumozyme has extremely high efficacy in sensitizing Spn3 to host immune mechanisms and that the enzyme has remarkable potential as a therapeutic. As part of Pneumotactix, LLC, our aims for this project are thus: Aim 1: Characterize the pharmacokinetics, potential immunogenicity and cytotoxicity of the enzyme. Aim 2: Elucidate the therapeutic value of the enzyme against Spn3 infection. In our published studies, we have used the native, full-length protein product to degrade the capsule of one virulent strain of Spn3 to great effect, and have further shown that this enzyme product does not induce cellular toxicity against host mammalian cells in both in vitro and in vivo experiments. Our immediate goals for this project are therefore to expand these studies to test pharmacokinetics, assess enzyme efficacy against additional strains of serotype 3 Spn, and perform continued experiments to confirm the therapeutic value of Pneumozyme and concurrent lack of off-target effects upon administration. The long-term goals for Pneumozyme are to establish it as a therapeutic agent by testing its efficacy in higher animal systems (i.e., primates) with significant sample numbers of Spn type 3 clinical isolates, and move towards clinical trials to develop this enzyme as a highly relevant therapeutic against this incredibly virulent pathogen.

项目摘要 肺炎链球菌（SPN）仍然是最致命的传染剂之一 每年有百万人死亡，并构成大多数继发感染 免疫功能低下的患者。与该病原体相关的主要毒力机制之一是 多糖胶囊决定了细菌的成功定植和感染 作为防御病原体杀死免疫介导的防御机制。 SPN血清型3（SPN3）是 被认为是SPN最具毒性的血清型之一，尽管当前的多价疫苗包括 针对SPN3的共轭靶标，SPN感染病例中该血清型的总比例实际上具有 在过去的十年中有所增加。此外，SPN3与抗生素耐药性的增加直接相关。 因此，SPN3影响的严重程度以及在设计疗法时产生的困难，不能 被夸大了。肺化（PN3Pase）是一种酶产物，我们以前已证明是高度的 有效降解3型SPN的囊囊多糖。我们的体外研究表明，治疗 带有肺化的SPN3将使细菌更容易受到吞噬细胞杀死机制的影响。 此外，用SPN3致命挑战的小鼠在用肺化治疗后可以救出： 100％接受肺化治疗生存的小鼠挑战率，而接受的小鼠接受了100％ 在SPN3致命挑战后的两天内，非活性酶或车辆死亡。这些结果表明肺化 在敏化SPN3方面具有极高的功效，以托管免疫机制，并且该酶具有显着的 潜力作为治疗性。作为Pneumotactix，LLC的一部分，我们对该项目的目标是： 目标1：表征酶的药代动力学，潜在的免疫原性和细胞毒性。 目标2：阐明酶对SPN3感染的治疗值。 在我们已发表的研究中，我们使用了本机，全长蛋白产品来降解一个 SPN3的有力菌株产生了很大的作用，并进一步表明该酶不会诱导细胞 在体外和体内实验中对宿主哺乳动物细胞的毒性。我们为此的直接目标 因此，项目将扩大这些研究以测试药代动力学，评估酶疗效 血清型3 SPN的其他菌株，并执行继续实验以确认 肺化和同时缺乏对给药的脱靶影响。长期目标 肺化应通过测试其在较高动物系统中的功效（即，即， 灵长类动物）具有大量SPN 3型临床分离株的样本数量，并朝着临床试验转向 将这种酶作为一种高度相关的治疗方法，以针对这种令人难以置信的毒性病原体。