Catalytic Antibody Approach to Flunitrazepam Degradation

描述（由申请人提供）： 该提案系统地探讨了降解氟硝西泮的催化抗体的免疫反应，氟硝西泮是一种滥用的苯二氮卓类药物，也涉及药物辅助性和/或身体攻击。氟硝西泮是一种中枢神经系统抑制剂，具有长期镇静催眠作用，也可诱导顺行性遗忘。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。两个半抗原构建在一个过渡态类似物的方法，针对氟硝西泮的1，2-酰胺水解，取代一个半抗原中的磷酰胺和磺酰胺在其他的酰胺。第三种半抗原靶向氟硝西泮的4，5-亚胺，用在生理pH下带负电荷的近端羧酸酯取代它。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正点电荷以启动亚胺水解。杂交技术用于产生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分析催化作用。主动和被动免疫具有治疗效用。

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.