Development of new agents that regulate cellular signal transduction

开发调节细胞信号转导的新药物

基本信息

批准号：
15208012
负责人：
OHIGASHI Hajime
金额：
$ 28.45万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15208012/
关键词：
aplysiatoxin bryostatin C1 domain diacylglycerol kinase indolactam phorbol ester protein kinase C tumor promoter 亜鉛フィンガー C1ドメインプロテインキナーゼC GFP RasGRP chimaerin Unc13 indolactam-V

项目摘要

Protein kinase C (PKC) isozymes are major receptors of tumor-promoting phorbol esters. They contain two cysteine-rich C1 domains (C1A, C1B), both of which are candidates for phorbol 12,13-dibutyrate (PDBu) binding sites. To investigate the mechanism of tumor promotion in molecular level, the C1 peptides corresponding to the C1 domains of all PKC isozymes were synthesized, and their dissociation constants for PDBu were measured. The resultant C1 peptide library was used to screen for new ligands with PKC isozyme and C1 domain selectivity to find that indolactam-V (IL-V) is a promising lead compound.We verified that the indole ring of IL-V could be involved in the CH/πinteraction with Pro-11 of the C1B domain of PKCδ using its mutant peptide, in which the CH/π interaction was inhibited by substitution of the hydrogen atom with a fluorine atom at position 4 of Pro-11. IL-V showed about a 10 times lower binding affinity to the mutant peptide compared to the wild-type peptide, suggesting th … More at the CH/π interaction could play a pivotal role on the binding of IL-V to the PKCδ C1B domain. On the other hand, benzolactam-V8 (BL-V8), with the benzene ring instead of the indole ring of IL-V, might lack the CH/π interaction. The low binding affinity of BL-V8 could be enhanced by the effective formation of the CH/n interaction as exemplified by the synthesis of naphtholactam-V8. Based on the difference among the CH/π interaction in PKC isozymes, 1-hexyl-indolactma-V10 and 8-octyl-benzolactam-V9 with potent selectivity for novel PKC isozymes (δ,ε,η,θ) that play significant roles in tumor promotion were developed. Moreover, a simplified analogue of tumor-promoting aplysiatoxin with less hydrophobicity was design-synthesized and shown to translocate PKCδ from cytosol to nuclear membrane like bryostatin 1 with anti-neoplastic activity.Recently, new phorbol ester receptors other than PKC (n-chimaerins, Unc-13s, and RasGRPs) have been reported. We unambiguously demonstrated that diacylglycerol kinase (DGK) γ and β are new targets of tumor-promoting phorbol esters by synthesizing the C1 peptides of all DGK isozymes. Less

蛋白质激酶C（PKC）同工酶是肿瘤 - 原酯的主要受体。测量所有PKC同工酶的解离常数PDBU。 IL-V的环可能与使用其突变肽与PKCδ的C1b域的pro-11进行CH/π互动，其中氢原子与氟原子原子在Pro-原子上抑制了CH/π相互作用11。与野生型肽相比，与突变酰胺的结合亲和力低约10倍，这表明在CH/π相互作用时更多的是在IL-V与PKC C1B结构域的结合中起关键作用苯甲酸苯甲酸苯甲酸苯酚（BL-V8），带有苯环而不是IL-V的吲哚环，CH/π相互作用可以通过有效形成CH/N来增强BL-V8的低结合。基于PKC同工酶中CH/π相互作用的差异，1-己基 - 异生酶C同工酶（δ，ε，η，θ）在CH/π相互作用中的差异所示例。此外，较少的肿瘤促进蛋白毒素的简化类似物是设计合成的，并显示了从胞质胶到核膜1的PKCδ，如抗肿瘤蛋白1，具有抗神经性活性。 -13s和rasgrps）我们明确地证明了二酰基乙二醇激酶（DGK）促进肿瘤的phorbol酯通过合成所有DGK同工酶的C1肽。