Antibody-based countermeasures against fentanyl and its analogues

基于抗体的芬太尼及其类似物对策

基本信息

批准号：
10015669
负责人：
Marco Pravetoni
金额：
$ 81.38万
依托单位：
UNIVERSITY OF MINNESOTA
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：
2020-09-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10015669
关键词：
Address Adjuvant Administrative Supplement Affinity Alfentanil Antibodies Antigens B-Lymphocytes Behavior Benchmarking Binding Biological Assay Bradycardia Brain Buprenorphine Canada Chemical Warfare Chemicals Cloning Contractor Development Drug Kinetics Europe Exposure to FDA approved Fab Immunoglobulins Family Fentanyl Formulation Future Generations Goals Half-Life Heroin Hospitalization Human Hybridomas Immunization Immunize Immunoglobulin Fragments In Vitro Industry Industry Standard Intellectual Property Lead Length Lymphocyte Macaca mulatta Magnetism Methadone Military Personnel Modeling Modification Monoclonal Antibodies Mus Naloxone Naltrexone North America Opioid Opioid Antagonist Opioid Receptor Oxycodone Passive Immunization Pharmaceutical Preparations Poisoning Polymers Process Public Health Rattus Receptors, Antigen, B-Cell Risk Risk Assessment Series Site Structure Sufentanil System Technology Testing Toxic effect Transgenic Mice United States Vaccines Ventilatory Depression X-Ray Crystallography analog base biocompatible polymer biodegradable polymer biophysical properties carfentanil chemical threat commercialization efficacy testing endogenous opioids humanized monoclonal antibodies humanized mouse improved in vivo lead optimization mass casualty medical countermeasure meetings mouse model next generation nonhuman primate opioid overdose polyclonal antibody polymerization preclinical development research clinical testing scale up screening

项目摘要

ABSTRACT. This project will develop monoclonal antibodies (mAb) against toxicity and lethality from accidental or deliberate exposure to fentanyl, carfentanil, acetylfentanil, and alfentanil. The US has seen dramatic increases in fatal opioid poisoning due to the widespread availability of fentanyl and fentanyl-like analogs. Because of the risk for accidental or deliberate mass casualty incidents, the fentanyl chemical family is listed in the DHS Chemical Threat Risk Assessment list. As a complementary adjuvant strategy to current medications for rescue of opioid poisoning (e.g., the opioid receptor antagonist naloxone), our team is developing mAb against fentanyl and its potent analogs to reduce toxicity and lethality in both civilian and defense applications. Our team has shown that antibody-based strategies effectively reduce opioid distribution to the brain as well as opioid-induced respiratory depression and bradycardia in mice and rats. Antibodies selectively target the intended opioid, but do not interfere with endogenous opioids nor with naloxone or other approved medications. Our team has identified lead first-generation mouse anti-fentanyl mAbs using an antigen-based enrichment strategy to isolate opioid-specific B cell lymphocytes from immunized mice to generate hybridomas, and mouse/human chimeric mAb for expression in a suitable mammalian system. The proposed U01 project will further optimize these leads through an iterative developmental plan. AIM1 focuses on optimization of humanized mAb against fentanyl, carfentanil, acetylfentanyl, and alfentanil by integrating humanization of lead mouse mAb and immunization of humanized or transgenic mice. The lead mAb will be used as a benchmark for selection of next-generation candidates. Leads will be identified for their affinity and selectivity in vitro, and for in vivo efficacy in reducing antinociception, respiratory depression and lethality in rats. The humanization process will be guided by biophysical characterization of mAb binding to fentanyl and its analogs by X-ray crystallography. AIM2 focuses on optimization of lead mAb (Fab or F(ab)2 fragments as alternative options) via polymer-based modifications to improve half-life and volume of distribution, and increase efficacy against fentanyl and its analogs. AIM3 will test efficacy of leads co-administered as a multicomponent mAb formulation in combination with opioid antagonists to provide enhanced protection against respiratory depression and lethality in rats challenged with fentanyl and its derivatives. To provide proof of scalability, AIM4 tests the efficacy of the lead humanized mAb in a non-human primate model of opioid-induced respiratory depression. Leads from previous aims will be synthesized at a larger scale at a contractor site. Finally, AIM5 focuses on drafting a regulatory path toward late-stage development and commercialization.

抽象的。该项目将开发单克隆抗体 (mAb)，以抵抗意外事故造成的毒性和致死性或故意接触芬太尼、卡芬太尼、乙酰芬太尼和阿芬太尼。美国出现了大幅增长由于芬太尼和芬太尼类似物的广泛使用，导致致命的阿片类药物中毒。因为由于存在意外或故意造成大规模伤亡事件的风险，芬太尼化学品家族被列入国土安全部 (DHS) 化学威胁风险评估清单。作为当前救援药物的补充辅助策略为了防止阿片类药物中毒（例如阿片类受体拮抗剂纳洛酮），我们的团队正在开发针对芬太尼的单克隆抗体及其有效的类似物，可降低民用和国防应用中的毒性和致命性。我们的团队有研究表明，基于抗体的策略可有效减少阿片类药物向大脑的分布以及阿片类药物引起的小鼠和大鼠的呼吸抑制和心动过缓。抗体选择性地针对预期的阿片类药物，但是不干扰内源性阿片类药物，也不干扰纳洛酮或其他批准的药物。我们的团队有使用基于抗原的富集策略分离出第一代小鼠抗芬太尼单克隆抗体来自免疫小鼠的阿片特异性 B 细胞淋巴细胞产生杂交瘤，以及小鼠/人嵌合体用于在合适的哺乳动物系统中表达的 mAb。拟议的U01项目将进一步优化这些线索通过迭代开发计划。 AIM1专注于针对芬太尼的人源化单克隆抗体的优化，卡芬太尼、乙酰芬太尼和阿芬太尼，通过整合先导小鼠单克隆抗体的人源化和免疫接种人源化或转基因小鼠。先导单克隆抗体将作为选择下一代单克隆抗体的基准候选人。将鉴定先导物的体外亲和力和选择性，以及体内减少的功效大鼠的镇痛、呼吸抑制和致死作用。人性化进程将遵循通过 X 射线晶体学表征 mAb 与芬太尼及其类似物结合的生物物理特征。 AIM2聚焦通过基于聚合物的修饰优化先导单克隆抗体（Fab 或 F(ab)2 片段作为替代选项）改善半衰期和分布体积，并提高针对芬太尼及其类似物的功效。 AIM3将测试作为多组分单克隆抗体制剂与阿片类拮抗剂联合给药的先导药物的功效为受到芬太尼挑战的大鼠提供针对呼吸抑制和致死的增强保护它的衍生物。为了提供可扩展性证明，AIM4 在非人类试验中测试了先导人源化 mAb 的功效阿片类药物引起的呼吸抑制的灵长类动物模型。先前目标的线索将在更大范围内进行综合承包商现场的规模。最后，AIM5 的重点是起草一条通往后期开发和开发的监管路径。商业化。