ATALYTIC ANTIBODIES AND COCAINE

这项拨款申请提出了开发克隆抗体的建议。 会将可卡因的水解物催化成非精神活性物质 复方淫羊藿甲酯。它的基础是有报道称，催化 具有高度选择性酯酶样活性的单抗具有 是通过用羧酸的磷酸盐类似物免疫小鼠而获得的 酯类化合物。该膦酸盐结构模拟酯的过渡态 水解液。因此，抗体的结合部位稳定了 过渡态，加速酯类的水解。一种磷酸盐类似物 的可卡因将被合成并与蛋白质偶联。老鼠会成为 免疫并制备杂交瘤。这些杂交瘤将在 用酶联免疫吸附测定法与人血清白蛋白类似物结合 磷酸盐半抗原。来自阳性杂交瘤的抗体将被检测 因为它们能够催化[~3H]-可卡因(苯基)的水解 标记的)到[~3H]-苯甲酸，将被提取和测量。两者都有 抗体的催化和选择性方面将是 调查过了。预计这项拟议的试验性研究将取得成功 将导致拨款申请，以详细研究 免疫原对选择性和催化速率的影响及免疫原的作用 动物被动免疫对可卡因药代动力学的影响 药效学。可卡因被认为是一种极好的模型药物 以检验催化抗体在药物中应用的总体可行性 虐待研究。作为合成酶，它们可以用来增强 了解药物代谢过程。它们提供了一种手段 操纵药物从体内的排泄率，并研究 这一比例对药理活性的影响。例如，催化 抗体可用于研究其药效学效应。 可卡因的迅速消耗，从而揭示了周围的因素 急性耐受和戒断。固定化催化抗体可以 用于“人工肝”模型的药代动力学模拟 实验。最终，这一概念可能会扩展到其他药物 滥用和其他反应途径。除了它们的基本价值之外 药理学研究，催化抗体最终可能会 潜在的戒毒或可能被应用于药物滥用 通过阻止或减轻压力来拒绝加固的处理 吸毒的心理影响。

This grant application proposes the development of monoclonal antibodies which will catalyze the hydrolysis of cocaine to the non-psychoactive compound ecgonine methyl ester. It is based on reports that catalytic monoclonal antibodies with highly selective esterase-like activity have been obtained by immunizing mice with phosphonate analogs of carboxylic esters. The phosphonate structure mimics the transition state for ester hydrolysis. Thus the binding site of the antibody stabilizes the transition state and accelerates ester hydrolysis. A phosphonate analog of cocaine will be synthesized and coupled to protein. Mice will be immunized and hybridomas prepared. These hybridomas will be screened in an enzyme-linked immunosorbent assay for binding to an analog of the phosphonate hapten. Antibodies from positive hybridomas will be tested for their ability to catalyze the hydrolysis of [3H]-cocaine (phenyl labeled) to [3H]-benzoic acid which will be extracted and measured. Both the catalytic and selectivity aspects of the antibodies will be investigated. It is expected that success in this proposed pilot study will lead to a grant application to study in detail the factors in the immunogen affecting selectivity and catalytic rate and the effects of passive immunization of animals on cocaine pharmacokinetics and pharmacodynamics. Cocaine is viewed as an excellent model drug with which to test the overall feasibility of catalytic antibodies in drug of abuse research. As synthetic enzymes, they may be used to enhance understanding of the drug metabolism process. They offer a means of manipulate the rate of removal of drug from the body and study the effects of that rate on pharmacological activity. For example, catalytic antibodies may be used to investigate the pharmacodynamic effects of rapid depletion of cocaine and thus shed light on the factors surrounding acute tolerance and withdrawal. Immobilized catalytic antibodies could be used in model "artificial livers" for pharmacokinetic modeling experiments. Eventually the concept could be expanded to other drugs of abuse and other routes of reaction. In addition to their value in basic pharmacological studies, catalytic antibodies might eventually have potential for detoxification or perhaps could be applied in drug abuse treatment to deny reinforcement by blocking or mitigating the psychological effects of drug taking.