Catalytic Antibody Approach to Flunitrazepam Degradation

氟硝西泮降解的催化抗体方法

• 批准号: 6931402
• 负责人: ANDREW P BROGAN
• 金额: $ 4.21万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6931402
• 关键词: abzyme; benzodiazepines; biotechnology; catalyst; combinatorial chemistry; drug abuse chemotherapy; drug metabolism; haptens; hybridomas; immunologic substance development /preparation; immunopharmacology; immunotherapy; laboratory mouse; phage display; postdoctoral investigator

DESCRIPTION (provided by applicant): This proposal systematically probes the immune response for catalytic antibodies to degrade flunitrazepam, an abused benzodiazepine also implicated in drug-assisted sexual and/or physical assault. Flunitrazepam is a central nervous system depressant with prolonged sedative-hypnotic effects that also induces anterograde amnesia. Three different flunitrazepam analog haptens are designed to exploit the known degradation pathways of the drug using established methods to generate catalytic antibodies. Two haptens are constructed in a transition-state analog approach, targeting 1,2-amide hydrolysis of flunitrazepam, replacing the amide with a phosphonamidate in one hapten and a sulfonamide in the other. A third hapten targets the 4,5-imine of flunitrazepam, replacing it with a proximal carboxylate that is negatively charged at physiological pH. During immunization this hapten will program a positive point charge in the antibody to initiate imine hydrolysis when flunitrazepam is given as a substrate. Hybridoma technology is used to generate monoclonal antibodies for all three haptens. Catalysis will be analyzed using the intrinsic chromophores of flunitrazepam and its degradation products. Active and passive immunizations possess therapeutic utility.

描述（由申请人提供）： 该提案系统地探讨了催化抗体降解氟硝西泮的免疫反应，氟硝西泮是一种滥用的苯二氮卓类药物，也与药物辅助的性和/或身体攻击有关。氟硝西泮是一种中枢神经系统抑制剂，具有长效镇静催眠作用，也会引起顺行性遗忘。设计了三种不同的氟硝西泮类似物半抗原，以利用已知的药物降解途径，使用已建立的方法产生催化抗体。采用过渡态类似方法构建了两个半抗原，针对氟硝西泮的 1,2-酰胺水解，在一个半抗原中用膦酰胺取代酰胺，在另一个半抗原中用磺酰胺取代酰胺。第三个半抗原针对氟硝西泮的 4,5-亚胺，用在生理 pH 值下带负电荷的近端羧酸盐取代它。在免疫过程中，当氟硝西泮作为底物时，该半抗原将在抗体中编程一个正点电荷以引发亚胺水解。杂交瘤技术用于生成所有三种半抗原的单克隆抗体。将使用氟硝西泮及其降解产物的固有发色团来分析催化作用。主动和被动免疫具有治疗效用。