Catalytic Antibody Approach to Flunitrazepam Degradation

描述(由申请人提供)： 这项建议系统地探索催化抗体降解氟硝西潘的免疫反应，氟硝西潘是一种滥用的苯二氮卓类药物，也与药物辅助性和/或身体攻击有关。氟硝西潘是一种中枢神经系统镇静剂，具有较长的镇静催眠作用，也会导致顺行性遗忘。三种不同的氟硝西潘类似物半抗原被设计用来利用药物的已知降解途径，使用现有的方法来产生催化抗体。以过渡态模拟方法构建了两个半抗原，目标是氟硝西潘的1，2-酰胺水解物，一个半抗原中的酰胺被亚磷酸酰胺取代，另一个半抗原中的磺胺被取代。第三个半抗原针对氟硝西潘的4，5-亚胺，用近端的羧酸盐取代，在生理pH下带负电荷。在免疫过程中，当给予氟硝西潘作为底物时，该半抗原将在抗体中编程正点电荷以启动亚胺的水解。杂交瘤技术被用来产生针对所有三种半抗原的单抗。将利用氟硝西潘及其降解产物的固有发色团来分析催化作用。主动免疫和被动免疫具有治疗作用。

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.