Chemical Probes to Discover the Lacosamide Drug Targets: Synthesis and Evaluation

描述(申请人提供)：这项研究项目旨在确定神经毒剂(R)-乳糖胺(LCM)的生物靶标。(R)-LCM是治疗癫痫和神经性疼痛的第三阶段临床试验中出现的一种有效的抗癫痫药物。药理学研究证明，LCM具有独特的活性，使其有别于已知的抗癫痫药物。基于这些发现，我们假设(R)-LCM与不同的蛋白质结合，具有低到中等的亲和力。对LCM作用机制的理解(S)将有助于增加我们对癫痫和疼痛障碍的理解，并允许合理开发新的临床药物。我们的第一个具体目标是设计和合成来自(R)-LCM的分子探针，称为亲和诱饵(AB)、化学报告(CR)和AB&CR。这些药物将在NINDS抗惊厥药物筛选项目的动物模型中进行抗惊厥活性评估。在第二个特定目标中，我们使用LCM AB&CR试剂来检查一组选定的蛋白质，这些蛋白质构成了(R)-LCM的相关靶点。我们的第三个具体目标是利用AB&CR试剂，使用基于亲和力的方法识别小鼠大脑中的(R)-LCM蛋白靶标。最后，在最后一个特定的目的中，我们使用mRNA展示和LCM AB&CR试剂来询问小鼠脑蛋白质组的药物功能位点。具体目标4将由北卡罗来纳大学教堂山的刘实验室进行。我们所有生物学研究的核心是构建LCM AB和CR试剂，这些试剂通过AB基团共价修饰靶标，然后通过CR基团和生物正交探针从生物混合物中移除。尽管尚不清楚，但抗癫痫药物(R)-乳糖胺的作用机制已被证明不同于其他抗癫痫药物。乳糖胺药物靶点的确定将为我们提供重要的、新的信息，了解癫痫和神经性疼痛的生物学机制。这些知识将使新的抗癫痫和神经病理性止痛药的开发成为可能。

DESCRIPTION (provided by applicant): This research project aims at identifying the biological targets of the neurological agent (R)-lacosamide (LCM). (R)-LCM is a potent anti-epileptic agent emerging from Phase III clinical trials for the treatment of epilepsy and neuropathic pain. The pharmacological studies document that LCM has a unique profile of activity that differentiates it from known anti-epileptic agents. Based on these findings, we hypothesize that (R)-LCM binds to different proteins, with low to modest affinity. The understanding of LCM's mechanism of action(s) will help increase our understanding of seizure and pain disorders, and permit the rational development of new clinical agents. Our first specific aim is the design and synthesis of molecular probes derived from (R)-LCM termed Affinity Bait (AB), Chemical Reporter (CR), and AB&CR. These agents will be evaluated for anticonvulsant activity in animal models at the NINDS Anticonvulsant Screening Project. In the second specific aim, we use the LCM AB&CR agents to examine a select panel of proteins, which constitute relevant targets for (R)-LCM. Our third specific aim utilizes the AB&CR agents to identify the (R)-LCM protein targets in the mouse brain using an affinity-based approach. Finally, in the last specific aim we interrogate the mouse brain proteome using mRNA display and the LCM AB&CR agents for sites of drug function. Specific Aim 4 will be conducted by the Liu laboratory at UNC-Chapel Hill. Central to all our biological studies are the construction of the LCM AB&CR agents that covalently modify the target through the AB group and then are removed from the biological mixture via the CR group and a bioorthogonal probe. Though still unknown, the mechanism of action of the anti-epileptic agent (R)-lacosamide has been shown to differ from other anticonvulsant drugs. The identification of lacosamide drug targets will provide us important, new information of the biological mechanisms underlying epilepsy and neuropathic pain. This knowledge will allow the rational development of new anti-epileptic and neuropathic pain agents.