Molecular basis of prostanoid receptor-mediated pathogenesis and its drug application

前列腺素受体介导发病机制的分子基础及其药物应用

• 批准号: 12557211
• 负责人: ICHIKAWA Atsushi
• 金额: $ 8.51万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557211/
• 关键词: prostaglandin; receptor subtypes; signal transduction; gene targeting; cycloxygenase-2; colon cancer; colitis; receptor blocker; 喘息; 遺伝子欠損マウス; 好中球; 分娩; シクロオキシゲナーゼ

Results obtained by Kyoto University Group(1) Elucidation of patho-physiological roles of prostanoids by using each receptor-deficient mice :We found that EP2 deficiency attenuated intestinal polyp formation both in number and size in-Apc KO mice. We also showed acceleration of COX-2, EP2 and VEGF expression in the intestinal polyps of wild-type mice, but faint expression of COX-2 or VEGF in those of EP2 and Ape double-knockout mice, suggesting that there exists positive feedback regulation between COX-2 and EP2. We introduced dextransulfate (DSS)-induced colitis model into EP-deficient mice. As a result, only EP4-deficient mice showed severe colitis upon 3%DSS treatment that did not induce significant colitisin wild-type mice. EP4 has been shown to promote epithelial regeneration and to inhibit activation of leukocytes and lymphocytes.(2) The new signal transduction pathway exerted by prostanoid receptor EP3 :EP3 has been shown to be coupled to Gi activity, inhibition of adenylate cyc … More lase. EP3 has multiple isoforms that are different in their C-terminal structure and in their agonist-dependent Gi activity. We found that EP3 receptors expressed in COS-7 cells showed augmentation of other receptor-stimulated Gs activity in an agonist-dependent manner. The superactivation of Gs by EP3 receptors was not affected by the treatment of pertussis toxin, suggesting that the superactivation is not Gi-mediated. The EP3 agonist-dependent supeactivation of Gs is observed in any type of mouse EP3 receptor isoforms, irrespective of C-termnal structure of EP3 receptor. This EP3 signaling may reflect some aspects of the physiological function of PGE2, such as pain sensation.]Results obtained by Ono Pharmaceutical Co. Group(3) Development of EP4 receptor-specific ligands :Improvement of EP4-receptor selectivity and the agonist activity by introduction of heteroatoms into the alpha chain of PGE1 was investigated. Of the compounds produced, 16-phenyl-omega-tetranor-3,7-dithiaPGE1 possessing moderate EP4-receptor selectivity and agonist activity, was identified as a new chemical lead for further optimization by modification of the aromatic moiety Based on the information, we also successfully generated an EP4-specific antagonist with a high selectivity. Less

京都大学课题组的研究结果：(1)利用各受体缺陷小鼠阐明前列腺素的病理生理作用：我们发现EP2缺乏在-APC KO小鼠肠息肉形成的数量和大小上都有所减少。野生型小鼠肠息肉组织中COX-2、EP2和血管内皮生长因子表达加速，而EP2和APE双基因敲除小鼠肠息肉组织中COX-2和VEGF表达较弱，提示COX-2和EP2之间存在正反馈调节。我们将葡萄糖转硫酸盐(DSS)诱导的结肠炎模型引入EP缺陷小鼠。结果，只有EP4基因缺陷的小鼠在3%的DSS治疗下表现出严重的结肠炎，而在野生型小鼠中没有引起明显的结肠炎。EP4可促进上皮再生，抑制白细胞和淋巴细胞的激活。(2)前列腺素受体EP3：EP3发挥的新的信号转导途径与胃肠道活动、抑制腺苷环化…有关再来点激光。EP3有多种不同的异构体，它们的C末端结构和激动剂依赖的GI活性不同。我们发现，在COS-7细胞中表达的EP3受体以激动剂依赖的方式表现出其他受体刺激的Gs活性的增强。百日咳毒素处理不影响EP3受体对Gs的过度激活，提示这种过度激活不是由Gi介导的。在任何类型的小鼠EP3受体亚型中都可以观察到EP3激动剂依赖的Gs的过度激活，而与EP3受体的C-末端结构无关。这种EP3信号可能反映了PGE2生理功能的某些方面，如痛觉。]小野制药公司获得的结果(3)EP4受体特异性配体的开发：通过将杂原子引入PGE1的阿尔法链，研究了EP4受体选择性和激动剂活性的改善。在所合成的化合物中，16-苯基-欧米伽-四氢-3，7-二硫-PGE1具有中等的EP4受体选择性和激动剂活性，被鉴定为一种新的化学先导化合物，通过对芳香族部分的修饰，我们还成功地合成了一种高选择性的EP4特异性拮抗剂。较少

项目成果

Ma H, Hara A, Xiao CY, et al.: "Increased bleeding tendency and decreased susceptibility to thromboembolism in mice lacking the prostaglandin E receptor subtype EP3"Circulation. 104. 1176-1180 (2001)

Ma H、Hara A、Xiao CY 等人：“缺乏前列腺素 E 受体亚型 EP3 的小鼠出血倾向增加，血栓栓塞的易感性降低”循环。

Hatae, N., et al.: "Augmentation of Receptor-Mediated Adenylyl Cyclase Activity by Gi-Coupled Prostaglandin Receptor Subtype EP3 in a Gβγ Subunit-Independent Manner"Biochem. Biophys. Res. Commun.. 290. 162-168 (2002)

Hatae，N.，等人：“Gi 偶联前列腺素受体亚型 EP3 以不依赖 Gβγ 亚基的方式增强受体介导的腺苷酸环化酶活性”Biochem. 290. 162-168 (2002) ）

Hatae, N., Yamaoka, K., Sugimoto, et al.: "Augmentation of Receptor-Mediated Adenylyl Cyclase Activity by Gi-Coupled Prostaglandin Receptor Subtype EP3 in a Gβγ Subunit-Independent Manner"Biochem.Biophys.Res.Commun.. 290. 162-168 (2002)

Hatae, N.、Yamaoka, K.、Sugimoto 等人：“Gi 偶联前列腺素受体亚型 EP3 以 Gβγ 亚基独立方式增强受体介导的腺苷酸环化酶活性”Biochem.Biophys.Res.Commun. 290. 162-168 (2002)

Croy,B.A., et al.: "Prolonged gestation dose not extend survival of uterine natural killer lymphocytes in mice deleted in the receptor for prostaglandin F_<2α>."J.Reprod.Immunol.. 46. 125-129 (2000)

Croy, B.A. 等人：“在前列腺素 F_<2α> 受体缺失的小鼠中，延长妊娠不会延长子宫自然杀伤淋巴细胞的存活时间。”J.Reprod.Immunol.. 46. 125-129 (2000)

Tsuboi,K., et al.: "Uterine expression of prostaglandin H_2 synthase in late pregnancy and during parturition in prostaglandin F receptor-deficient mice"Endocrinology. 141. 315-324 (2000)

Tsuboi,K., et al.：“前列腺素 F 受体缺陷小鼠妊娠晚期和分娩过程中前列腺素 H_2 合酶的子宫表达”内分泌学。