A study for drug-discovery based on physiological actions of prostanoid receptors

基于前列腺素受体生理作用的药物发现研究

• 批准号: 10557222
• 负责人: ICHIKAWA Atsushi
• 金额: $ 8.64万
• 依托单位: KYOTO 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-10557222/
• 关键词: prostaglandin receptor; receptor subtype-selective agonist; G protein; prostaglandin receptor-deficient mice; ligand-receptor interaction; ovulation; fever; ductus arteriosus; EP3サブタイプ受容体; EP2受容体欠損マウス; 受精; 卵丘細胞; プロスタグランジン; PGE受容体欠損マウス; PGF受容

We established mice deficient in each type of prostanoid receptors, and clarified physiological roles of prostanoids by examining phenotypes in each knockout mice : FP-deficient mice fail to deliver their pups due to persistent production of progesterone in ovaries, indicating that PGF2α plays a role in luteolysis required for normal parturition. IP-deficient mice showed increased tendency of thrombosis, reduced responses of inflammation, and did not show acetic acid-induced writhing responses, indicating that PG12 participates in promotion of inflammation and pain sensation. EP3-deficent mice did not show febrile responses to both endogenous and exogenous pyrogen; PGE2 mediates pyrogen-induced fever generation via EP3. EP4-deficient mice died within three days after birth due to patency of ductus arteriosus. PGE2 plays an essential role in neonatal closure of ductus arteriosus via EP4. EP2-deficent female mice showed reduced fertility due to impaired ovulation and fertilization; PGE2 … More contributes to ovulation and fertilization through stimulation of cumulus expansion via EP2 and the resultant increase in cAMP level. Through these studies, we demonstrated importance of receptor subtype-specific activities.Furthermore, by the analysis of mutant EP3 receptor expressed in CHO cells generated by the method of site-directed mutagenesis, we found that ionic interaction between plus charge of arginine-residue (Arg-306 in mouse EP3 receptor) and minus charge of free carboxyl-residue in ligands is essential for the activation of Gs and Gq proteins, but hydrogen interaction in these residues is enough for the activation of Gi protein. Based on these findings, we developed four agonists for EP receptor subtype (EP1 : ONO-DI-004, EP2 : ONO-AE1-259, EP3 : ONO-EA-248, EP4 : ONO-AE1-329) and an EP1-specific antagonist (ONO-8711) which are excellent in their receptor-specificity. These compounds have potential to be developed as powerful drugs with more specific therapeutic effects and less side effects. Less

我们建立了每种类型前列腺素受体缺陷的小鼠，并通过检查每种敲除小鼠的表型来阐明前列腺素的生理作用：FP缺陷小鼠由于卵巢中持续产生孕酮而无法分娩幼崽，表明PGF 2 α在正常分娩所需的黄体溶解中发挥作用。IP缺陷小鼠表现出血栓形成倾向增加，炎症反应减少，并且没有表现出乙酸诱导的扭体反应，表明PG 12参与促进炎症和疼痛感觉。EP 3缺陷小鼠对内源性和外源性热原均未显示发热反应; PGE 2通过EP 3介导热原诱导的发热产生。EP 4缺陷小鼠在出生后三天内由于动脉导管的通畅而死亡。PGE 2通过EP 4在新生儿动脉导管闭合中起重要作用。缺乏EP 2的雌性小鼠由于排卵和受精受损而表现出生育力降低; ...更多信息 通过EP 2刺激卵丘扩张并由此增加cAMP水平，促进排卵和受精。通过这些研究，我们证明了受体亚型特异性活性的重要性，并通过对定点突变方法产生的CHO细胞表达的突变型EP 3受体的分析，我们发现，正电荷的精氨酸残基与正电荷的精氨酸残基之间的离子相互作用，（小鼠EP 3受体中的Arg-306）和配体中游离羧基残基的负电荷对于Gs和Gq蛋白的活化是必需的，但这些残基中的氢相互作用足以激活Gi蛋白。基于这些发现，我们开发了四种EP受体亚型的激动剂（EP 1：ONO-DI-004，EP 2：ONO-AE 1 -259，EP 3：ONO-EA-248，EP 4：ONO-AE 1 -329）和一种EP 1特异性拮抗剂（ONO-8711），它们具有优异的受体特异性。这些化合物有潜力被开发为具有更特异性治疗效果和更少副作用的强效药物。少

项目成果

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Hasekawa, H., et al.：“Madin-Darby 犬肾细胞中 Rho 的跨上皮电阻和细胞旁通透性的相反调节”。《生物化学杂志》274・30（1999）。

Hasegawa,H.,et al.: "Opposite Regulation of Transepithelial Electrical Resistance and Paracellular Permeability by Rho in Madin-Darby Canine Kidney Cells"J.Biol.Chem.. 274・30. 20982-20988 (1999)

Hasekawa, H.等人：“Madin-Darby犬肾细胞中Rho对跨上皮电阻和细胞旁通透性的相反调节”J.Biol.Chem.274・30(1999)。

Takeuchi K.,et al.: "Impaired duodenal bicarbonate secretion and mucosal integrity in mice lacking prostaglandin E-receptor subtype EP(3)"Gastroenterology. 117(5). 1128-1135 (1999)

Takeuchi K. 等人：“缺乏前列腺素 E 受体亚型 EP(3) 的小鼠十二指肠碳酸氢盐分泌和粘膜完整性受损”胃肠病学。

Ushikubi,F.et al.: "Impaired febrile response in mice lacking the prostaglandin E receptor subtype EP3." Nature. 395. 281-284 (1998)

Ushikubi, F. 等人：“缺乏前列腺素 E 受体亚型 EP3 的小鼠发热反应受损。”