A Novel Probiotic Drug Delivery Platform for Oral Administration of Protein-Based Therapies

用于口服蛋白质疗法的新型益生菌药物递送平台

• 批准号: 10544376
• 负责人: Christian Furlan Freguia
• 金额: $ 28.25万
• 依托单位: RISE THERAPEUTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2024-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10544376
• 关键词: Adoption; Animal Model; Arthritis; Bacteria; Biochemical Pathway; Biological Products; Biological Response Modifier Therapy; Biomass; Bioreactors; Carbohydrates; Carbon; Caustics; Cell Density; Cell Respiration; Cells; Clinical Trials; Cytochromes; DNA; Data; Development; Disease; Dose; Drug Delivery Systems; Engineering; Escherichia coli; Event; Evolution; Exhibits; Fermentation; Food Preservation; Gases; Genes; Genetic Engineering; Genetic Transcription; Gold; Gram-Positive Bacteria; Growth; Gut Mucosa; Half-Life; Heme; Human; Immune; Immune system; Immunologic Receptors; Immunotherapeutic agent; In Situ; Industrialization; Infrastructure; Intestines; Lactococcus; Lactococcus lactis; Lead; Malignant Neoplasms; Mediating; Metabolic Pathway; Metabolism; Molecular; Oral; Oral Administration; Oxidases; Oxygen; Pathway interactions; Performance; Pharmaceutical Preparations; Phase; Play; Probiotics; Process; Production; Productivity; Property; Protein Secretion; Proteins; Recombinants; Regulatory Pathway; Repression; Respiration; Role; Safety; Site; Sjogren' s Syndrome; Speed; Stress; System; Technology; Therapeutic; Therapeutic Agents; Toxic effect; Traction; Translating; bacterial metabolism; base; carbohydrate metabolism; cost; density; drug candidate; gut microbiome; host microbiome; improved; manufacturability; metabolic respiration; microbial; novel; novel therapeutics; preclinical study; prevent; product development; programs; protein expression; protein purification; prototype; research clinical testing; respiratory; sugar; synthetic biology; therapeutic protein

Project Summary Oral delivery of protein-based drugs (‘biologics’) would vastly improve and simplify our approach to treat and prevent disease1-4. However, biologics suffer from low stability and short half-life when administered in oral form and are sensitive to the caustic conditions of the intestinal tract5-7. Oral delivery of biologics via bacteria can circumvent these hurdles. Compared to conventional drug delivery systems, bacteria exhibit the advantages of in situ production of biologicals, targeting ability, and amenability to different payloads. By leveraging gut microbiome-host immune cell intertwining, bacteria delivery of therapeutics can also lead to modulation of the host immune response8-10. Rise Therapeutics has pioneered the developed of novel recombinant strains of the probiotic Lactococcus lactis demonstrating tremendous proof-of-concept for the approach in over 15 animal models. In these studies, oral delivery of our recombinant probiotics enables targeted engagement of specific receptors of the immune system to rebalance the host immune repertoire. Enabled by our in-house GMP manufacturing infrastructure, these recombinant strains are now entering human clinical testing. Lactococcus (L.) lactis is one of the most promising bacterial platforms for biologics delivery. L. lactis is a non- pathogenic, GRAS-designated, Gram-positive bacterium with an extraordinary safety profile in humans, including use in genetically modified forms to delivery therapeutic protein. In addition to oral delivery of biologics, microbial systems are a hallmark of industrial production of recombinant proteins11. Engineered L. lactis has also been employed to express heterologous proteins for industrial applications. While E. coli is the gold standard, Gram positive bacteria, like L. lactis are gaining traction due to much simpler purification processes and important proteins` biochemical pathways not supported by Gram negative strains 12, 13. However, L. lactis platform suffers from manufacturing limitations, where cellular bioreactor densities top off far below levels require for some commercial purposes. This limitation increases cost of goods, prevents use of higher doses in humans, and curb its use for industrial protein expression applications. Achieving high yield densities of pure and stable recombinant bacteria is essential to enabling commercially viable and successful product development. To solve this challenge, we will use synthetic biology approaches to modify key growth regulatory pathways to improve manufacturability of our promising L. lactis platform. L. lactis adapts to environmental circumstances to survive. In a bioreactor setting, L. lactis has a controlled metabolism that permits the utilization of certain type of sugars. When under stress, and event that typically occurs at late stage of fermentation, L. lactis switches to an aerobic metabolism, which is detrimental to the bacteria survival. In this application, we proposed to engineer a new L. lactis chassis with improved metabolic and respiration capacities to augment and improve bacterial fermentation and cellular biomass.

项目摘要 口服基于蛋白质的药物（“生物制剂”）将大大改善和简化我们治疗和治疗癌症的方法。 预防疾病1 -4.然而，生物制剂在口服给药时具有低稳定性和短半衰期的缺点。 形成并对肠道的腐蚀性条件敏感5 -7。通过细菌口服递送生物制剂 可以绕过这些障碍。与传统的药物递送系统相比，细菌表现出 生物制剂的原位生产、靶向能力和对不同有效载荷的适应性的优点。通过 利用肠道微生物组-宿主免疫细胞交织，细菌递送治疗剂也可导致 调节宿主免疫应答8 -10。Rise Therapeutics开创了新颖的 益生菌乳酸乳球菌的重组菌株证明了 在超过15种动物模型中。在这些研究中，口服我们的重组益生菌， 免疫系统的特异性受体的靶向接合以重新平衡宿主免疫库。 通过我们内部的GMP生产基础设施，这些重组菌株现在正在进入 人体临床试验 乳球菌属（L.）乳酸菌是用于生物制剂递送的最有前途的细菌平台之一。L.乳酸是一种非- 致病性、GRAS指定的革兰氏阳性细菌，在人体内具有非凡的安全性， 包括以遗传修饰的形式用于递送治疗性蛋白质。除了口服给药 生物制剂、微生物系统是重组蛋白工业生产的标志11。工程湖 乳酸菌也被用于表达工业应用的异源蛋白质。而E.大肠杆菌是 金标准，革兰氏阳性细菌，如L.由于更简单的提纯， 革兰氏阴性菌株不支持的过程和重要蛋白质的生化途径12，13。 然而，L. lactis平台受到制造限制，其中细胞生物反应器密度远远超过 低于某些商业用途所需的水平。这种限制增加了商品的成本，阻止了 更高的剂量在人类中，并遏制其用于工业蛋白质表达应用。实现高产 纯的和稳定的重组细菌的密度对于实现商业上可行的和成功的 产品开发 为了解决这一挑战，我们将使用合成生物学方法来修改关键的生长调节途径， 提高我们有前途的L的可制造性。lactis平台L.乳酸菌适应环境条件， 生存在生物反应器中，L.乳酸菌具有受控的代谢， 糖。当受到胁迫时，以及通常发生在发酵后期的事件，L. lactis切换到 有氧代谢，这对细菌的生存是有害的。在本申请中，我们建议 设计一种新的L.乳酸底盘与改善的代谢和呼吸能力，以增加和改善 细菌发酵和细胞生物量。