Trace charactethation of neurosteroids to develop a new antipsychotic agent targeting neurosteroidogenesis

追踪神经类固醇的特征，开发一种针对神经类固醇生成的新型抗精神病药

• 批准号: 18590031
• 负责人: HIGASHI Tatsuya
• 金额: $ 2.61万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590031/
• 关键词: Neurosteroid; LC-MS; Derivatization; Stress; Rat brain; Antipsychotic agent; 神経活性アンドロゲン; コルチコイド; 検出指向誘導体化; GABA_A受容体; 抗不安・鎮静作用; エタノール

Neurosteroids bind to the γ-aminobutyric acid type A (GABA_A) receptors with a high affinity, positively modulate the action of GABA at these receptors and thus elicit marked anesthetic, sedative and anxiolytic effects. The quantitative analysis of the endogenous neurosteroids in the brain and serum is very important for characterization of their physiological functions and the mechanism by which they affect the brain functions. The brain and serum assays of the neurosteroids can also contribute to the development of new antipsychotic agents targeting neurosteroidogenesis. Based on this background information, sensitive liquid chromatography-tandem mass spectrometric methods that enable the quantification of trace neurosteroids (5α-redduced pregnane neurosteroids, 3-oxo-4-ene-neurosteroids, neuroavtive androgens and tetrahydrocorticosterone isomers) in rat brain and serum were developed and validated. Some methods employed derivatization procedures, which were very effective in increas … More ing the detection responses of steroids.New discoveries from the animal studies using the developed methods were as follows. 1) The brain allopregnanolone, epiallopregnanolone and 5α-dihydroprogesterone levels are rapidly elevated by immobilization stress and ethanol administration. Epiallopregnanolone is the brain-specific product. 2) The stress-induced level changes in the brain testosterone and androstenedione (androstane steroids) are much lower than those of progesterone and its metabolites (pregnane steroids). 3) Most of the brain testosterone is derived from the peripheral organs, while a fair amount of 5α-dihydrotestosterone is continuously synthesized in the brain. The GABAergic neuroactive androgen, 5α-androstane-3α,17β-diol, is always present in the male rat brain and its level is not influenced by the stress. 4) 3α,5α-Tetrahydrocorticosterone is present in the brain of both male and female rats after being subjected to the immobilization stress, but there is a sex difference in its brain level. Less

神经甾体以高亲和力结合γ-氨基丁酸A （GABA_A）受体，正调节GABA对这些受体的作用，从而引起显著的麻醉、镇静和抗焦虑作用。脑和血清中内源性神经类固醇的定量分析对于表征其生理功能及其影响脑功能的机制具有重要意义。脑和血清神经类固醇的测定也有助于开发新的针对神经类固醇生成的抗精神病药物。基于这一背景信息，建立并验证了灵敏液相色谱-串联质谱法定量测定大鼠脑和血清中微量神经甾体（5α-还原孕激素、3-氧-4-烯-神经甾体、神经活性雄激素和四氢皮质酮异构体）的方法。有些方法采用衍生化程序，可以有效地提高类固醇的检测反应。使用开发的方法进行动物实验的新发现如下。1)固定应激和乙醇处理可使大鼠脑内异孕酮、表异孕酮和5α-二氢孕酮水平迅速升高。表异孕酮是脑特异性产物。2)应激诱导的脑内睾酮和雄烯二酮（雄烯酮类固醇）水平变化远低于孕酮及其代谢产物（孕激素类固醇）。3)脑内睾酮主要来源于外周器官，同时也有相当数量的5α-二氢睾酮在脑内不断合成。gaba能神经活性雄激素5α-雄甾烷-3α，17β-二醇在雄性大鼠脑中始终存在，且其水平不受应激的影响。4)固定应激后，雌雄大鼠脑内均存在3α，5α-四氢皮质酮，但其脑内水平存在性别差异。少

项目成果

LC/電子捕獲APCI-MSによるラット脳内corticosterone代謝物の同定

通过 LC/电子捕获 APCI-MS 鉴定大鼠脑中的皮质酮代谢物

• 发表时间: 2007
• 作者: 横井宏之;東 達也;島田和武
• 通讯作者: 島田和武

Analysis of stress-induced changes in rat brain neuroactive steroid levels using LC/MS coupled with derivatization

使用 LC/MS 结合衍生化分析应激引起的大鼠脑神经活性类固醇水平变化

• 发表时间: 2007
• 作者: T. Higashi;A. Nagahama;Y. Ninomiya;K. Shimada
• 通讯作者: K. Shimada

Studies on neurosteroids XIX -: Development and validation of liquid chromatography-tandem mass spectrometric method for determination of 5α-reduced pregnane-type neurosteroids in rat brain and serum

• DOI: 10.1016/j.jchromb.2006.10.036
• 发表时间: 2007-04-01
• 期刊: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
• 影响因子: 3
• 作者: Higashi, Tatsuya;Nagahama, Akihiro;Shimada, Kazutake
• 通讯作者: Shimada, Kazutake

ピリジル基の導入及びニトロフェニルヒドラゾン形成に基づくステロイドのESI-MS用高感度検出法の開発

基于吡啶基引入和硝基苯腙形成的类固醇ESI-MS高灵敏检测方法的开发

• 发表时间: 2007
• 作者: 西尾 忠;東 達也;島田和武
• 通讯作者: 島田和武

ステロイドの水酸基を標的としたLC/ESI-MS用誘導体化試薬の開発

开发针对类固醇羟基的 LC/ESI-MS 衍生化试剂

• 发表时间: 2006
• 作者: 西尾 忠;田中 淳;東 達也;島田和武
• 通讯作者: 島田和武