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.

项目概要/摘要 基于伊立替康的化疗广泛用于治疗各种类型的转移性癌症，然而， 严重药物引起的损伤或胃肠道（GI）毒性，尤其是严重迟发性腹泻（SDOD） SN-38（伊立替康的活性代谢物）诱导的细胞毒性，限制了该药物的临床应用。伊立替康诱导 SDOD 发生率会导致住院导致的医疗费用增加以及生活质量差和 癌症患者的治疗结果。目前，约 15% 的患者尚无有效治疗方法 接受伊立替康治疗的患者出现无法控制的 SDOD 症状，主要是由于他们无能力 解毒结肠中的 SN-38。 SN-38 介导的肠道毒性被发现在患者中更为严重 具有尿苷二磷酸糖基转移酶 1A1 (UGT1A1) 多态性，描绘了关键作用 结肠 UGT1A1 预防 SN-38 诱导的 SDOD。减少 SN-38 结肠的各种方法 已经尝试过暴露但没有取得太大成功，并且 SDOD 仍然无法控制剂量限制性毒性 伊立替康用于成人和儿童癌症患者。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 蒺藜苜蓿可以有效地代谢典型的人类 UGT1A1 底物，包括 SN-38， 体外测定。这催生了生物工程共生细菌大肠杆菌（EC）的新颖和创新概念。 过表达植物 UGT71G1 以产生一种可以解毒 SN-38 的药物解毒细菌 (DDB)。在这个STTR中 项目中，我们建议生成概念验证临床前证据，以支持开发安全、 有效且可追溯的 DDB 作为一种新型活体生物治疗产品 (LBP)，可缓解和/或预防 SN- 38介导的肠道毒性。在这里，我们将通过生物工程改造出一种具有良好人类肠道功能的共生大肠杆菌菌株 定殖潜力，过表达绿色标记的 UGT71G1 (U71G1*n) 最活跃变体 荧光蛋白 (GFP)，将装在保护性胶囊中直接递送至结肠，以有效 SN-38 糖基化为 SN-38-葡萄糖。为了实现这一目标，提出的具体目标是 1） 可追踪且更活跃的药物解毒细菌 EC_U71G1*n（活性变体）对 SN-38 进行解毒，2) 开发可追踪且高活性冻干 DDB 的结肠优化胶囊输送，以及 3) 评估 2 种结肠递送 DDB 在伊立替康诱导的 SDOD 大鼠模型中的功效。主动 DDB 设计有 pUGT 生物催化剂（用于 SN-38 的解毒）和 GFP（用于追踪人类肠道有效定植） 用于治疗效果监测的粪便）将加速癌症患者从 SDOD 中恢复，并导致 临床中伊立替康诱发的 SDOD 的有效管理。该项目的成功将提供 Sanarentero 拥有开发针对 SN-38 的可追踪 DDB 所需的概念验证证据 II 期 SBIR/STTR 应用中的临床前和 IND 支持研究。小说 LBP 将充当 治疗 SDOD 的补充疗法，应改善生活质量甚至治疗结果 对于转移性癌症患者。一旦这项工作取得成功，我们计划采用相同的方法来利用 不同的植物 UGT 用于灭活其他胃肠道毒性药物、异生物质、污染物或其有毒代谢物 通过开发药物特异性 DDB 来治疗结肠癌。