IRREVERSIBLE ANTAGONISTS OF COCAINE AND OTHER STIMULANTS

这里提出的工作目的是合成化合物 这将对抗强化和诱发精神病的特性 通过不可逆地结合到兴奋剂识别来识别可卡因兴奋剂 多巴胺转运蛋白上的位点。这个方法是基于之前的 Metaphit 的发现，一种酰化苯环利啶衍生物， 体外不可逆地抑制末端以及转运 多巴胺神经末梢。 在体内，Metaphit 完全阻断 多动症通常在服用可卡因后出现 其他兴奋剂。 可卡因和其他兴奋剂的潜在不可逆拮抗剂将 通过向已知化合物添加反应性亲电基团来合成 是与多巴胺结合的兴奋剂的有效抑制剂 运输者以及运输过程本身。化合物 选择进行修饰的是马吲哚，“GBR”系列芳基 1,4-二烷（烯）基哌嗪、哌醋甲酯和多巴胺本身。的 要引入其结构中的反应基团是 异硫氰酸酯、溴乙酰胺、马来酰亚胺和氟磺酰基部分。 新合成的化合物将与 Fourphit（一种 由外部合作者向我们提供的 Metaphit 的异构体） 与多巴胺转运蛋白不可逆相互作用的能力。这将 通过测量（1）它们对兴奋剂的抑制活性来确定 [3H]哌醋甲酯放射受体测定中的识别位点和 (2) 它们不可逆地阻断[3H]多巴胺摄取到突触体中的能力。 多巴胺转运蛋白对去甲肾上腺素的选择性 将确定血清素转运蛋白。发现化合物不可逆 在一定程度上抑制兴奋剂结合和多巴胺摄取 然后将在体内测试其选择性的行为影响。 化合物的活性将在三个单独的行为中进行检查 旨在评估其对抗的潜力的范式 强化和急性和慢性精神病诱发特性 人类兴奋剂。显示行为活性的化合物将 然后进行体外测试其阻断[3H]哌醋甲酯的能力 结合和[3H]多巴胺摄取。这将需要检查 [3H]哌醋甲酯结合和脑组织体外摄取多巴胺 取自体内用这些化合物处理过的大鼠。代谢物 使用兴奋剂治疗后还将测量多巴胺 拮抗剂，以便关联刺激拮抗剂的特性 合成的化合物及其对多巴胺周转的影响。这个 研究将使我们能够开发兴奋剂药物的潜在拮抗剂 的虐待。这也为最终的隔离奠定了基础。 多巴胺转运复合物的纯化。

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.