NEUROPSYCHOPHARMACOLOGY OF CYCLIC AMP PDE INHIBITORS

描述(改编自申请人的摘要)：宽泛的、长期的 本研究的目的是阐明PDE4在行为学中的作用 调节和介导PDE4抑制剂的AD效应。 首先，我们将进行实验，以确定哪些PDE4亚型 (PDE4A、PDE4B、PDE4D)参与AD样行为的调节 效应，以及其他中枢神经系统效应。这将通过以下方式实现 新开发的PDE4亚型选择性抑制剂的疗效比较 在两个行为任务的大鼠中，行为在DRL 72秒下保持 时间表和两级罗利普兰药物识别程序，以及他们的 重组大鼠PDE4A、PDE4B、PDE4C和PDE4D的抑制作用。相关 实验将检验反义的行为效应。 针对PDE4A变体(PDE4A1和PDE4A5)的寡核苷酸(ODN) 它们被认为参与了AD样行为的调节 效果。第二，将进行实验，以确定 行为效应主要是由抑制物相互作用介导的。 PDE4上的低亲和力或高亲和力罗利普兰结合位点。这将是 通过比较PDE4抑制剂的行为效应和 他们与这些网站互动的亲和力。第三，实验将 以确定哪些PDE4子类型与所选的 大脑中受受体刺激的腺苷环化酶。这将会实现的 通过确定PDE4亚型选择性抑制剂的效果，以及 针对PDE4A变异体的反义ODN，关于水解性 腺苷-3‘，5’-环一磷酸(CAMP) β-1和β-2肾上腺素能或D1多巴胺能受体刺激的腺酰化 环化酶在大鼠脑片中的表达。第四，实验将通过以下方式检验该方式 突触前神经支配和环磷酸腺苷调节神经元中的PDE4。这 将通过确定被抑制的突触发生的影响来实现， 以突触素II为靶点的反义ODN诱导，或循环中的变化 腺苷环化酶抑制剂或非氢化环状AMP诱导的AMP 类似物在原代培养神经元中PDE4变异体的发育 大鼠大脑皮层。 拟议的实验结果将显示PDE4的哪些亚型 阿尔茨海默病的主要介质及PDE4的其他行为效应 抑制剂；此外，低-和 将建立高亲和力的罗利普兰结合位点。最后，PDE4 与特定信号转导系统相关的变体和 突触前神经支配和环磷酸腺苷调节这些神经元的方式 这些联系将会被阐明。提高了对功能的理解 而PDE4在大脑中的药理学将开始阐明它在 为抑郁症的病理生理学及改善提供合理依据 药物疗法。

DESCRIPTION (Adapted from applicant's abstract): The broad, long-term objective of this proposal is to elucidate the role of PDE4 in behavioral regulation and in the mediation of the AD effects of PDE4 inhibitors. First, experiments will be carried our to determine which PDE4 subtypes (PDE4A, PDE4B, PDE4D) are involved in the mediation of AD-like behavioral effects, as well as other CNS effects. This will be accomplished by comparing the effects of newly developed PDE4 subtype-selective inhibitors in rats in two behavioral tasks, behavior maintained under a DRL 72-sec schedule and a two-lever rolipram drug discrimination procedure, with their inhibition of recombinant rat PDE4A, PDE4B, PDE4C, and PDE4D. Related experiments will examine the behavioral effects of antisense oligodeoxynucleotides (ODNs) targeted to PDE4A variants (PDE4A1 and PDE4A5) which are hypothesized to be involved in the mediation of AD-like behavioral effects. Second, experiments will be carried out to determine if the behavioral effects are mediated predominantly by inhibitor interaction with the low- or high-affinity rolipram binding sites on PDE4. This will be accomplished by comparing the behavioral effects of PDE4 inhibitors with their affinities for interacting with these sites. Third, experiments will be carried out to determine which PDE4 subtypes are associated with selected receptor-stimulated adenyl cyclase in the brain. This will be accomplished by determining the effects of PDE4 subtype-selective inhibitors, as well as antisense ODNs targeted to PDE4A variants, on the hydrolysis of adenosine-3',5'-cyclic monophosphate (cyclic AMP) formed by stimulation of beta-1 and beta-2 adrenergic or D1 dopaminergic receptor-stimulated adenylyl cyclase in rat brain slices. Fourth, experiments will examine the manner by which presynaptic innervation and cyclic AMP regulate PDE4 in neurons. This will be accomplished by determining the effects of inhibited synaptogenesis, induced with antisense ODNs targeted to synapsin II, or changes in cyclic AMP, induced by an adenylyl cyclase inhibitor or by nonhydrozable cyclic AMP analogs, on the development of PDE4 variants in primary neuronal cultures of rat cerebral cortex. The results of the proposed experiments will show which subtypes of PDE4 are the predominant mediators of the AD and other behavioral effects of PDE4 inhibitors; in addition the relative importance of the low- and high-affinity rolipram binding sites will be established. Finally, the PDE4 variants associated with particular signal transduction systems and the manner by which presynaptic innervation and cyclic AMP regulate these associations will be elucidated. The improved understanding of the function and pharmacology of PDE4 in the brain will begin to clarify its role in the pathophysiology of depression and provide a rational basis for improved pharmacotherapy.