Development of drugs which open or close ATP-sensitive KィイD1+ィエD1channels

开发打开或关闭 ATP 敏感 KiiD1+iED1 通道的药物

• 批准号: 10557002
• 负责人: INAGAKI Nobuya
• 金额: $ 7.1万
• 依托单位: AKITA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10557002/
• 关键词: sulfonylurea; chimera; G-protein; thiazolidinedione; ATP-sensitive KィイD1+ィエD1channel; substania nigra; knockout mouse; ischemia; 神経細胞; 黒質網様部; ATP感受性K^+チャネル; インスリン; スルホニル尿素; 膵β細胞; 心筋細胞

ATP-sensitive KィイD1+ィエD1(KィイD2ATPィエD2) channels are key molecules which link the cell's metabolic status to its membrane potential. We have determined that pancreatic β-cell KィイD2ATPィエD2 channel comprises the inward rectifier Kir6.2 and the sulfonylurea receptor SUR1 subunits, while the cardiac KィイD2ATPィエD2 channel comprises Kir6.2 and SUR2A, an isoform of SUR1. Our purpose in this study is to develop the drugs which open or close these channels specifically. Our results are as follows.# We have determined the regions which confer the sensitivities of the KィイD2ATPィエD2 channels to the sulfonylurea and the KィイD2ATPィエD2 channel opener diazoxide, by analyzing a series of chimeras between SUR1 and SUR2.# We have investigated the effects of G-protein on the reconstituted KィイD2ATPィエD2 channels. We have shown that G-protein α subunit directly regulates the KィイD2ATPィエD2 channel activity, and that the regulation is different between β-cell and cardiac KィイD2ATPィエD2 channels, suggesting that the d … More ifference is determined by distinct SUR subunits.# We have investigated the effects of the thiazolidinedione derivatives, which are known to improve insulin resistance, on the reconstituted KィイD2ATPィエD2 channels. Troglitazone inhibited β-cell and cardiac KィイD2ATPィエD2 channels in a dose dependent manner; it inhibited the former at 30-100 μM and the late at 3 μM. This suggests that troglitazone modulates the various cellular functions including insulin secretion and muscle contraction especially under ischemic condition.# It has been known that KィイD2ATPィエD2 channels are expressed in brain, and are especially abundant in substantial nigra. We have studied the electrophysiological properties of the KィイD2ATPィエD2 channel activity in the acutely dissociated neurons from substantia nigra pars reticulate (SNr). The pharmacological properties of the channel are similar to those of the β-cell KィイD2ATPィエD2 channel. KィイD2ATPィエD2 channel knockout mice generated by targeting the Kir6.2 gene (established in Professor Seino's lab in Chiba University), in which KィイD2ATPィエD2 channel activity is absent in SNr, are vulnerable to ischemia, suggesting that KィイD2ATPィエD2 channels play an important role in protection against an ischemic insult. Less

ATP 敏感的 KィイD1+ィエD1(KィイD2ATPィエD2) 通道是将细胞代谢状态与其膜电位联系起来的关键分子。我们确定胰腺β细胞KiiD2ATPィエD2通道包含内向整流器Kir6.2和磺酰脲受体SUR1亚基，而心脏KiiD2ATPィエD2通道包含Kir6.2和SUR2A（SUR1的同工型）。我们这项研究的目的是开发专门打开或关闭这些通道的药物。我们的结果如下。#通过分析SUR1和SUR2之间的一系列嵌合体，我们确定了KィイD2ATPィエD2通道对磺酰脲类和KィイD2ATPィエD2通道开放剂二氮嗪敏感的区域。#我们研究了G蛋白对重组体的影响。 KィイD2ATPィエD2 通道。我们已经证明，G 蛋白 α 亚基直接调节 KィイD2ATPィエD2 通道活性，并且 β 细胞和心脏 KィイD2ATPィエD2 通道之间的调节是不同的，这表明差异是由不同的 SUR 亚基决定的。# 我们研究了已知可改善胰岛素抵抗的噻唑烷二酮衍生物对重组细胞的影响。 KィイD2ATPィエD2 通道。曲格列酮以剂量依赖性方式抑制 β 细胞和心脏 KiiD2ATPィD2 通道；它在 30-100 μM 下抑制前者，在 3 μM 下抑制后者。这表明曲格列酮调节各种细胞功能，包括胰岛素分泌和肌肉收缩，尤其是在缺血条件下。#众所周知，KiiD2ATPィD2通道在大脑中表达，并且在黑质中尤其丰富。我们研究了黑质网状部 (SNr) 急性分离神经元中 KィイD2ATPィエD2 通道活性的电生理特性。该通道的药理学特性与 β 细胞 KィイD2ATPィエD2 通道的药理学特性相似。通过靶向 Kir6.2 基因（在千叶大学 Seino 教授实验室建立）产生的 KィイD2ATPィエD2 通道敲除小鼠，其中 SNr 中缺乏 KィイD2ATPィエD2 通道活性，很容易发生缺血，这表明 KィイD2ATPィエD2 通道在预防缺血性损伤方面发挥着重要作用。较少的

项目成果

Nakata, M. and Inagaki, N.: "Molecular mechanism of insulin recretion."Mebio. 15. 24-29 (1998)

Nakata, M. 和 Inagaki, N.：“胰岛素分泌的分子机制”。Mebio。

Yamada k.,Nakata M.,Horimoto N.,Saito M.,Matuoka H.and Inagaki N.: "Measurement of glucose uptake and intracellular calcium concentration in single, living pancreatic β-cells"J. Biol. Chem.. (in press). (2000)

Yamada K.、Nakata M.、Horimoto N.、Saito M.、Matuoka H. 和 Inagaki N.：“单个活胰腺 β 细胞中葡萄糖摄取和细胞内钙浓度的测量”J. 2000)

Sanchez, J.A., Gonoi, I., Inagaki, N., Katada, T., and Seino, S.: "Modulation of reconstituted ATP-sensitive K^+ channels by GTP-binding proteins." J.Physiol.507. 315-324 (1998)

Sanchez, J.A.、Gonoi, I.、Inagaki, N.、Katada, T. 和 Seino, S.：“GTP 结合蛋白对重建 ATP 敏感 K^ 通道的调节”。

Suzuki,M.,Fujikura,K.,Kotake,K.,Inagaki,N.,et al.,: "Immuno-localization of sulfonylurea receptor 1 in rat pancreas."Diabetologia. 42. 1204-1211 (1999)

Suzuki,M.、Fujikura,K.、Kotake,K.、Inagaki,N.等人：“大鼠胰腺中磺酰脲受体 1 的免疫定位。”糖尿病学。

Sunaga, Y., Inagaki, N., et al.: "Troglitazone but not pioglitazone affects ATP-sensitive KィイD1+ィエD1 channel activity."Eur. J. Pharmacol.. 381. 71-76 (1999)

Sunaga, Y., Inagaki, N. 等人：“曲格列酮影响 ATP 敏感的 KiD1+KiD1 通道活性。”Eur. J. Pharmacol.. 381. 71-76 (1999)