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

基本信息

批准号：
10080253
负责人：
Gary Fanger
金额：
$ 95.78万
依托单位：
RISE THERAPEUTICS, LLC
依托单位国家：
美国
项目类别：
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/10080253
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 vaccine delivery

项目摘要

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.

项目概要该项目的目标是开发一种新药 R-4329，用于预防艰难梭菌 (C.) 感染 (CDI) 是一种经过改造的益生菌，可重组表达并口服输送独特的微生物蛋白。被称为分泌性抗原 A (SagA)，我们的团队证明它可以预防多种肠道感染，包括艰难梭菌，通过增强肠膜屏障的完整性/功能1, 2。艰难梭菌是一种普遍存在的厌氧革兰氏阳性细菌，可以形成孢子并具有高度抗性环境压力和一线抗生素，例如克林霉素7。艰难梭菌的致病菌株。毒素（A 和 B）会损害肠上皮细胞，导致炎性结肠炎、严重腹泻、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。自 2000 年以来，当个体的内源性微生物群发生改变或严重减少时，进行抗生素治疗。 CDI发病率从不足15万例增加到超过25万例，已成为全球最严重的疾病之一在美国，最严重的医院获得性感染的治疗经济负担超过 10 亿美元7。一旦艰难梭菌在结肠中定居，就很难用最后的抗生素来根除（万古霉素），从而导致 CDI 和炎症性结肠炎终生复发。治疗 CDI 的方法是粪便微生物移植 (FMT) 疗法，但这种方法是异质且定义不明确的治疗方法，其安全性仍然是一个问题，需要 FDA 进行特殊处理迫切需要新的、明确的治疗方法来预防和治疗复发性 CDI。口服益生菌为控制 CDI 和乳酸菌菌株（嗜酸菌、干酪菌、罗伊氏菌、植物菌）已被探索用于抑制肠道病原体感染并减轻抗生素相关性不幸的是，其作用机制很难理解，但对腹泻的有益作用有限。然而，这些最近的研究表明，益生菌的临床用途有所改善。事实上，我们最近的研究结果可能有助于控制 CDI。发表在《科学》杂志上的文章表明，对益生菌进行工程或“重新编程”可以重组表达 SagA 对肠道感染（包括 CDI2）具有更强的保护功能。我们的目标是开发一种新型益生菌菌株，以口服形式提供重组 SagA（称为 R-4329）该药物可自然发挥作用，诱导保护性调节信号。申请旨在以成功为基础，推动 R-4329 走向临床测试的具体目标。包括：1) 优化 R-4329 上游工艺开发并创建 GMP 主细胞库，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 剂量优化和基于生物标志物的小鼠 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 归档。