IRREVERSIBLE ANTAGONISTS OF COCAINE AND OTHER STIMULANTS

本文提出的工作目的是合成化合物 这会对抗增强和精神病诱导特性 通过不可逆地与兴奋剂识别结合， 多巴胺转运体上的位点。这种方法是基于以前的 Metaphit是一种酰化苯环苯胺衍生物， 在体外不可逆地抑制两个终端以及运输 多巴胺神经末梢。在体内，Metaphit完全阻断 在服用可卡因后通常会出现活动过度， 其他兴奋剂。 可卡因和其他兴奋剂的潜在不可逆拮抗剂将 通过向已知的化合物中加入反应性亲电子基团来合成 有效抑制兴奋剂与多巴胺的结合 运输工具，以及运输过程本身。化合物 选择进行修饰的是马吲哚，即“GBR”系列芳基化合物 1，4-二（烯）基哌嗪、哌甲酯和多巴胺本身。的 引入其结构中的反应性基团是 异硫氰酸酯、溴乙酰胺、马来酰亚胺和氟磺酰基部分。 新合成的化合物将被筛选，沿着与Fourphit（a Metaphit的异构体由外部合作者提供给我们）， 与多巴胺转运蛋白不可逆地相互作用的能力。这将 通过测量（1）它们对刺激物的抑制活性来确定 [3 H]哌甲酯放射性受体测定中的识别位点和（2） 其不可逆地阻断[3 H]多巴胺摄取进入突触体的能力。 多巴胺转运蛋白对去甲肾上腺素的选择性， 将测定血清素转运体。发现化合物不可逆地 抑制兴奋剂结合和多巴胺摄取， 然后将在体内测试它们的行为效应的选择性。 化合物的活性将在三个单独的行为测试中进行检查。 设计的范例，以评估其对抗 强化和急性和慢性精神病诱导特性 人体内的兴奋剂表现出行为活性的化合物将 然后离体测试它们阻断[3 H]哌甲酯的能力， 结合和[3 H]多巴胺摄取。这将需要审查 哌甲酯与脑组织多巴胺的体外结合和摄取 取自体内用这些化合物处理的大鼠。代谢产物 多巴胺也将在用兴奋剂治疗后测量 拮抗剂，以关联的刺激拮抗剂的性质， 合成的化合物及其对多巴胺周转的影响。这 研究将使我们能够开发出潜在的兴奋剂拮抗剂 虐待它也为最终的孤立奠定了基础， 纯化多巴胺转运复合物。

The purpose of the work proposed here is to synthesize compounds which will antagonize the reinforcing and psychosis-inducing properties of cocaine stimulants by binding irreversibly to the stimulant recognition site on the dopamine transporter. This approach is based on previous findings with Metaphit, an acylating phencylidine derivative, which in vitro irreversibly inhibits both the terminals as well as the transport of dopamine nerve terminal.In vivo, Metaphit completely blocks the hyperactivity normally seen following the administration of cocaine and other stimulants. Potential irreversible antagonists of cocaine and other stimulants will be synthesized by adding reactive electrophilic groups to compounds known to be effective inhibitors both of stimulant binding to the dopamine transporter, as well as of the transport process itself. The compounds selected for modification are mazindol, the "GBR' series of aryl 1,4-dialk(en)ylpiperazines, methylphenidate, and dopamine itself. The reactive groups to be introduced into their structures are the isothiocyanate, bromoacetamide, maleimide, and fluorosulfonyl moieties. The newly synthesized compounds will be screened, along with Fourphit (a isomer of Metaphit provided to us by an outside collaborator) for their ability to interact irreversibly with the dopamine transporter. This will be determined by measuring (1) their inhibitory activity at the stimulant recognition site in the [3H]methylphenidate radioreceptor assay and (2) their ability to irreversibly block [3H]dopamine uptake into synaptosomes. Selectivity for the dopamine transporter over the norepinephrine and serotonin transporter will be determined. Compounds found to irreversibly inhibit stimulant binding and dopamine uptake with some degree of selectivity will then be tested in vivo for their behavioral effects. Activity of the compounds will be examined in three separate behavioral paradigms designed to evaluate their potential to antagonize the reinforcing and acute and chronic psychosis-inducing properties of stimulant agents in humans. Compounds which show behavioral activity will then be tested Ex vivo for their ability to block [3H]methylphenidate binding and [3H]dopamine uptake. This will entail examining [3H]methylphenidate binding and dopamine uptake in vitro from brain tissue taken from rats treated with these compounds in vivo. Metabolites of dopamine will also be measured following treatment with the stimulant antagonists in order to correlate the stimulant antagonist properties of the synthesized compounds with their effect on dopamine turnover. This research will enable us to develop potential antagonists of stimulant drugs of abuse. It also lays the groundwork for the eventual isolation and purification of the dopamine transport complex.