A Novel Engineered Probiotic for the Prevention of Clostridium Difficile Infection

用于预防艰难梭菌感染的新型工程益生菌

• 批准号: 10198740
• 负责人: Gary Fanger
• 金额: $ 91.38万
• 依托单位: RISE THERAPEUTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10198740
• 关键词: Abdominal Pain; Affect; Anaerobic Bacteria; Animals; Anti-Infective Agents; Antibiotic Therapy; Antibiotics; Antigens; Bacteria; Biological; Biological Assay; Biological Markers; Biomass; Cell Wall; Chemicals; Cleaved cell; Clinical; Clinical Research; Clostridium; Clostridium difficile; Colitis; Colon; Culture Media; Data; Development; Diarrhea; Dose; Economic Burden; Elements; Engineered Probiotics; Engineering; Enteral; Enterococcus; Enterococcus faecium; Environment; Environmental Risk Factor; Enzymes; Epithelial Cells; Food; Future; Genetic Engineering; Goals; Gram-Positive Bacteria; Growth; Homeostasis; Host resistance; Human; Hydrolase; Individual; Industry; Infection; Infection prevention; Infectious Agent; Inflammatory; Intestines; Investments; Journals; Lactobacillus acidophilus; Lactococcus; Mediating; Medical; Membrane; Modeling; Mucous Membrane; Mus; N-Acetylmuramoyl-L-alanine Amidase; Nosocomial Infections; Nutrient; Oral; Pathogenesis; Pathogenicity; Patients; Pattern recognition receptor; Peptides; Peptidoglycan; Pharmaceutical Preparations; Pharmacodynamics; Pharmacologic Substance; Phase; Prevention; Probiotics; Process; Production; Property; Proteins; Publishing; Recombinants; Recovery; Recurrence; Relapse; Research Design; Resistance; Resort; Safety; Salmonella; Salmonella typhimurium; Schedule; Science; Signal Transduction; Small Business Innovation Research Grant; Stomach; Stress; Symptoms; Therapeutic; Toxicology; Toxin; Vancomycin; alternative treatment; base; biomarker identification; cell bank; cost; dosage; drug development; enteric infection; enteric pathogen; fecal transplantation; flu; high risk; host microbiota; improved; in vivo; innovation; interest; intestinal epithelium; lactic acid bacteria; manufacturing scale-up; microbial; microbiota; novel; novel therapeutics; pharmacokinetics and pharmacodynamics; prevent; research clinical testing; residence; success; transplantation therapy; vaccine delivery

Project Summary The goal of this project is to develop a novel drug, R-4329, for prevention of Clostridium (C.) difficile infection (CDI). R-4329 is a probiotic engineered to recombinantly express and orally deliver a unique microbial protein known as secreted antigen A (SagA) which our team showed can protect against multiple enteric infections, including C. difficile, by enhancing the integrity/functionality of the gut membrane barrier1, 2. C. difficile is a ubiquitous anaerobic Gram-positive bacterium that can sporulate and become highly resistant to environment stresses and frontline antibiotics such as clindamycin7. Pathogenic strains of C. difficile secret toxins (A and B) that damage intestinal epithelial cells, resulting in inflammatory colitis, severe diarrhea, abdominal pain, flu-like symptoms, and possible death7. C. difficile infection (CDI) often occurs following antibiotic-treatment when the endogenous microflora of individuals is altered or severely reduced. Since 2000, the rate of CDI has increased from below 150,000 to over 250,000 cases and has been become one of the most significant hospital-acquired infections with an economic burden of over $1 billion to treat in the U.S.7. Once C. difficile establishes residence in the colon, it is difficult to eradicate with last resort antibiotics (vancomycin) and consequently results in lifelong relapses of CDI and inflammatory colitis. An effective approach to treating CDI has been fecal microbiota transplant (FMT) therapy but this approach is heterogeneous and a poorly defined therapeutic, for which its safety is still a concern requiring special FDA approval8. New and well-defined therapeutics are desperately needed to prevent and treat recurrent CDI. Oral probiotics offer an exciting alternative to manage CDI and strains of Lactobacillus (acidophilus, casei, reuteri, plantarum) have been explored to inhibit enteric pathogen infection and mitigate antibiotic-associated diarrhea9. Unfortunately, mechanism of action has been difficult to understand with modest beneficial effects in the context of their clinical utility10. However, these recent studies suggest that improved probiotics with directed and targeted anti-infective functionality may be useful at controlling CDI. Indeed, our recent results published in the journal Science demonstrate that engineering or ‘reprogramming’ of probiotics to recombinantly express SagA yields a more protective functionality against enteric infections, including CDI2. Our goal is to develop a novel probiotic strain to deliver recombinant SagA (referred to as R-4329) as an orally administered drug that functions naturally to induce protective regulatory signals. This Phase II SBIR application is intended to build upon success and advance R-4329 towards clinical testing. The specific aims are: 1) optimize R-4329 upstream process development and create a GMP master cell bank, 2) manufacture R-4329 and develop GLP analytical product release/stability assays, 3) complete R-4329 dose optimization and biomarker-based PD studies in mice, 4) perform R-4329 GLP toxicology studies in mice to support our IND filing.

项目摘要 该项目的目的是开发一种新型药物R-4329，以预防梭状芽胞杆菌（C.）艰难梭菌感染 （CDI）。 R-4329是一种益生菌，旨在重组表达和口服独特的微生物蛋白 我们的团队表现出的被称为分泌的抗原A（Saga）可以预防多个发起人， 包括艰难梭菌，通过增强肠膜屏障的完整性/功能1，2。 艰难梭菌是一种无处不在的厌氧革兰氏阳性细菌，可以孢子孢子并高度抗性 环境压力和前线抗生素，例如克林霉素7。艰难梭菌秘密的致病菌株 毒素（A和B）损害肠上皮细胞，导致炎症性结肠炎，严重的腹泻， 腹痛，流感样症状和可能的死亡7。艰难梭菌感染（CDI）经常发生 当个体的内源性微生物群体被改变或严重降低时，抗生素治疗。自2000年以来 CDI的比率已从15万以下增加到250,000例，并且是其中之一 在美国7中，最重要的医院可获得感染，经济伯宁超过10亿美元。 一旦艰难梭菌在结肠中建立了居住，很难用持续的抗生素进行放射素 （万古霉素），因此导致CDI和炎性结肠炎的终生复发。有效 治疗CDI的方法是粪便菌群移植（FMT）疗法，但这种方法是 异质性且定义较差的疗法，其安全仍然是需要特殊FDA的问题 批准8。迫切需要新的且定义明确的治疗剂来预防和治疗复发性CDI。 口服益生菌为管理CDI和乳酸菌菌株提供了一种令人兴奋的替代方法 已探索了Reuteri，Plantarum）以抑制肠道病原体感染并减轻抗生素相关 腹泻9。不幸的是，行动机理在很难理解的情况下很难理解 他们的临床公用事业的背景10。但是，这些最近的研究表明，改善了益生菌 定向和靶向的抗感染功能可能有助于控制CDI。确实，我们最近的结果 发表在《科学杂志》上表明，益生菌的工程或“重编程” 重组表达的传奇产生了针对促进感染的更具保护功能，包括CDI2。 我们的目标是开发一种新颖的益生菌菌株，以提供重组传奇（称为R-4329）作为口服 施用自然作用可诱导保护性调节信号的药物。这个II期SBIR 应用旨在以成功为基础，并将R-4329推进临床测试。具体目标 是：1）优化R-4329上游流程开发并创建GMP大师库，2）制造 R-4329并开发GLP分析产品释放/稳定性测定，3）完成R-4329剂量优化 在小鼠中基于生物标志物的PD研究，4）在小鼠中进行R-4329 GLP毒理学研究以支持我们的IND 备案。