Development of Drug Detoxifying Bacteria for Chemotherapy Induced Gut Injury

开发用于化疗引起的肠道损伤的药物解毒细菌

• 批准号: 10252721
• 负责人: MING HU
• 金额: $ 40万
• 依托单位: SANARENTERO LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-23 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10252721
• 关键词: Adult; Attenuated; Bacteria; Biological Response Modifier Therapy; Biomedical Engineering; Cancer Patient; Clinic; Clinical; Collaborations; Colon; Complementary therapies; Diarrhea; Disseminated Malignant Neoplasm; Distal part of ileum; Dose; Dose-Limiting; Drug Metabolic Detoxication; Encapsulated; Engineering; Enzymes; Escherichia coli; FDA approved; Feces; Fluorescence; Fluorescent Protein Tracings; Freeze Drying; Gastric Acid; Genetic Polymorphism; Glucose; Glucosyltransferases; Goals; Green Fluorescent Proteins; Health Care Costs; Hospitalization; Hour; Human; Incidence; Injury; Intestines; Lead; Malignant Childhood Neoplasm; Measures; Mediating; Medicago truncatula; Medical; Modeling; Monitor; Patients; Pharmaceutical Preparations; Phase; Plant Proteins; Plants; Polymers; Prevention; Process; Proteins; Quality of life; Rattus; Recombinants; Recovery; Research; Role; SN-38; Safety; Series; Site; Small Business Innovation Research Grant; Small Business Technology Transfer Research; Symptoms; System; Technology; Texas; Toxic effect; Treatment Cost; Treatment Efficacy; UGT1A1 gene; Universities; Uridine Diphosphate; Variant; Xenobiotics; base; capsule; chemotherapy; clinical application; commensal bacteria; design; drug development; effective therapy; efficacy evaluation; gastrointestinal; glycosylation; glycosyltransferase; gut colonization; gut microbiota; improved; in vitro Assay; in vivo; innovation; irinotecan; novel; novel therapeutics; overexpression; pollutant; pre-clinical; preclinical efficacy; prevent; screening; success; therapy outcome

PROJECT SUMMARY/ ABSTRACT Irinotecan-based chemotherapy is widely used for the treatment of various types of metastatic cancers, however, severe drug-induced injury or gastrointestinal (GI) toxicity, especially severe delayed-onset diarrhea (SDOD) induced by SN-38 (an active metabolite of irinotecan), limits its clinical application of the drug. Irinotecan-induced SDOD incidences result in increased healthcare cost due to hospitalization, and poor quality of life and therapeutic outcomes for the cancer patients. Currently, there is no effective treatment available for about ~15% of patients receiving irinotecan therapy, who suffer unmanageable SDOD symptoms, mainly due to their inability to detoxify the SN-38 in the colon. SN-38 mediated intestinal toxicity was found to be more severe in patients with uridine diphosphate glycosyltransferases 1A1 (UGT1A1) polymorphism, delineating the critical role of colonic UGT1A1 in the prevention of SN-38 induced SDOD. Various approaches to reduce SN-38 colonic exposure has been tried without much success, and SDOD remains unmanageable dose-limiting toxicity of irinotecan in adult and pediatric cancer patients. Preliminary research of our academic research collaborators at the University of North Texas and the University of Houston discovered glucosyltransferases from plant Medicago truncatula that could efficiently metabolize typical human UGT1A1 substrates, including SN-38, in an in vitro assay. This led to the novel and innovative concept of bioengineering commensal bacteria E.coli (EC) for overexpressing plant UGT71G1 to create a drug detoxifying bacteria (DDB) that can detoxify SN-38. In this STTR project, we propose to generate proof-of-concept preclinical evidence in support of developing a safe, efficacious, and traceable DDB as a novel live biotherapeutic product (LBP) to alleviate and/or prevent the SN- 38-mediated intestinal toxicity. Here, we will bioengineer a commensal E.coli strain, with good human gut colonization potential, to overexpress the most active variant of UGT71G1 (U71G1*n) tagged with a green fluorescent protein (GFP), which will be delivered in a protective capsule directly to the colon for the effective glycosylation of SN-38 to SN-38-glucose. To achieve this goal, the proposed specific aims are 1) to develop traceable and more active drug detoxifying bacteria EC_U71G1*n (active variant) to detoxify SN-38, 2) to develop a colon-optimized capsule delivery of traceable and highly active lyophilized DDBs, and 3) to evaluate the efficacy of 2 colon-delivered DDBs in irinotecan induced SDOD rat model. An active DDB engineered with pUGT biocatalyst (for the detoxification of SN-38) and GFP (for tracing the effective gut colonization in human feces for therapeutic efficacy monitoring) will accelerate the recovery of cancer patients from SDOD, and lead to the effective management of irinotecan-induced SDOD in clinics. The success of this project will provide Sanarentero with the proof-of-concept evidence needed for developing an SN-38-targeted traceable DDB for preclinical and IND-enabling studies in a phase-II SBIR/STTR application. The novel LBP will act as a complementary therapy to treat SDOD, which should improve the quality of life and even therapeutic outcomes for metastatic cancer patients. Once successful in this endeavor, we plan to apply the same approach to utilize different plant UGTs for inactivating other GI-toxic drugs, xenobiotics, pollutants, or their toxic metabolites in the colon by developing drug-specific DDB.

项目摘要/摘要 基于伊立替康的化学疗法被广泛用于治疗各种类型的转移性癌症 严重的药物诱导的损伤或胃肠道（GI）毒性，尤其是严重的延迟发作的腹泻（SDOD） 由SN-38（一种活跃的伊立替康代谢产物）诱导，限制了其对药物的临床应用。伊立齿引起的 SDOD发病率导致由于住院和生活质量差而增加医疗保健成本， 癌症患者的治疗结果。目前，尚无大约15％的有效治疗方法 接受伊立替康治疗的患者，患有难以控制的SDOD症状的患者主要是由于他们的无能为力 在结肠中排毒SN-38。发现患者的SN-38介导的肠毒性更为严重 与尿苷二磷酸糖基转移酶1A1（UGT1A1）多态性一起描述 结肠UGT1A1预防SN-38诱导SDOD。减少SN-38结肠的各种方法 接触没有太大的成功，SDOD仍然是无法控制的剂量限制毒性 成人和小儿癌患者的伊立康。我们的学术研究合作者的初步研究 北德克萨斯大学和休斯顿大学从植物中发现了葡萄糖基转移酶 Medicago truncatula可以有效地代谢典型的人类UGT1A1底物，包括SN-38 体外测定。这导致了生物工程共生细菌E.Coli（EC）的新颖而创新的概念 过表达的植物UGT71G1可产生可以对SN-38排毒的药物排毒细菌（DDB）。在这个sttr中 项目，我们建议生成概念证明的临床前证据，以支持开发安全， 有效且可追溯的DDB作为一种新型的实时生物治疗产品（LBP），以减轻和/或防止SN- 38介导的肠毒性。在这里，我们将以良好的人类肠道菌株生物工程师 殖民电位，过表达用绿色标记的UGT71G1（U71G1*N）的最活跃的变体 荧光蛋白（GFP），该蛋白将直接在保护胶囊中递送至结肠 SN-38对Sn-38-葡萄糖的糖基化。为了实现这一目标，拟议的具体目标是1） 可追溯，更活跃的药物排毒EC_U71G1*n（活性变体）以排毒Sn-38，2） 开发可追溯且高度活跃的冻干DDB的结肠优化的胶囊递送，3）评估 2个结肠递送DDB在伊立替康诱导的SDOD大鼠模型中的功效。由 PUGT Biocatalyst（用于SN-38的排毒）和GFP（用于追踪人类的有效肠道化定植 治疗疗效监测的粪便）将加速从SDOD中恢复癌症患者，并导致 伊立替康诱导的SDOD在诊所中的有效管理。该项目的成功将提供 Sanarentero具有开发以SN-38为目标可追溯DDB所需的概念验证证据 在II期SBIR/STTR应用中，临床前和辅助研究。小说LBP将充当 治疗SDOD的互补疗法，这应该改善生活质量甚至治疗结果 用于转移性癌症患者。一旦成功完成这项工作，我们计划采用相同的方法来利用 不同的植物UGT用于使其他GI毒性药物，异种生物，污染物或其毒性代谢产物失活 结肠通过开发药物特异性DDB。