Synthesis and PKC binding of conformationally restricted indolactams by aza-Claisen rearrangement

通过氮杂克莱森重排合成构象限制的吲哚内酰胺并与 PKC 结合

• 批准号: 08660137
• 负责人: IRIE Kazuhiro
• 金额: $ 1.28万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08660137/
• 关键词: aza-Claisen rearrangement; teleocidin; indolactam; tumor promoter; protein kinase C; conformation

Protein kinase C (PKC) is a key enzyme family involved in cellular signal transduction and tumor promotion. It consists of a catalytic domain for protein phosphorylation and a regulatory domain which binds tumor promoters like phorbol esters and teleocidins. The tumor promoter-binding site in PKC is a cysteine-rich domain (CRD) at the N-terminal regulatory region. With the discovery of eleven PKC isozymes, increasing importance is placed on isozyme specific analysis of function. Isozyme-selective activators or inhibitors represent powerful tools to analyze the role of PKC and become new medicinal leads for diseases related to PKC activation like the vascular complications by hyperglycemia.However, the development of such compounds has been hampered since pure PKC isozymes are not easily obtainable from natural sources. We have synthesized the CRD's of PKCgamma, delta, and eta consisting of ca.50 amino acids by solid phase strategy to establish the models of native PKCs. These peptides were efficiently folded upon zinc treatment to produce PKC regulatory domain surrogates that bind [^3H] phorbol 12,13-dibutyrate (PDBu) with high affinity comparable to native PKC itself, suggesting that these peptides serve as effective models for native PKCgamma, delta, and eta.Teleocidins and its core structure (-) -indolactam-V are most promising as lead compounds for isozyme selective activation or inhibition of PKC since they are chemically very stable and easily synthesized. Using the PKC surrogate peptides, we have identified two new indolactam derivatives with high PKCgamma CRD selectivity based on our strategy for the design of conformationally restricted indolactam derivatives by aza-Claisen rearrangement. Bridge formation between position 5 and 13 of (-) -indolactam-V was proved to be one of the most effective methods to fix the molecule to the active conformation and to develop new PKC activators with high isozyme selectivity.

蛋白激酶C (PKC) 是参与细胞信号转导和肿瘤促进的关键酶家族。它由蛋白质磷酸化的催化结构域和结合佛波酯和teleocidins等肿瘤启动子的调节结构域组成。 PKC 中的肿瘤启动子结合位点是 N 端调控区的富含半胱氨酸结构域 (CRD)。随着 11 种 PKC 同工酶的发现，同工酶特异性功能分析变得越来越重要。同工酶选择性激活剂或抑制剂是分析 PKC 作用的有力工具，并成为治疗与 PKC 激活相关的疾病（例如高血糖引起的血管并发症）的新药物先导物。然而，由于不容易从天然来源获得纯 PKC 同工酶，此类化合物的开发受到阻碍。我们通过固相策略合成了由约50个氨基酸组成的PKCgamma、delta和eta的CRD，以建立天然PKCs模型。这些肽在锌处理后有效折叠，产生结合 [^3H] 佛波醇 12,13-二丁酸酯 (PDBu) 的 PKC 调节结构域替代物，其亲和力与天然 PKC 本身相当，表明这些肽可作为天然 PKCgamma、delta 和 eta 的有效模型。Teleocidins 及其核心结构 (-) -indolactam-V 最有希望作为先导药物 用于同工酶选择性激活或抑制 PKC 的化合物，因为它们化学性质非常稳定且易于合成。基于我们通过氮杂克莱森重排设计构象限制的吲哚内酰胺衍生物的策略，使用 PKC 替代肽，我们鉴定了两种具有高 PKCgamma CRD 选择性的新吲哚内酰胺衍生物。 (-)-吲哚内酰胺-V的5位和13位之间的桥接被证明是将分子固定到活性构象并开发具有高同工酶选择性的新型PKC激活剂的最有效方法之一。

项目成果

Kazuhiro Irie: "Synthesis and biological activities of new conformationally restricted analogues of (-) -indolactam-V : elucidation of the biologically active conformation of the tumor-promoting teleocidins" J.Am.Chem.Soc.118・44. 10733-10743 (1996)

Kazuhiro Irie：“(-)-indolactam-V 的新构象限制类似物的合成和生物活性：促进肿瘤的 teleocidins 的生物活性构象的阐明”J.Am.Chem.Soc.118・44。 (1996)

Kazuhiro Irie et al.: "Comparison of chemical characteristics of the first and the second cysteine-rich domains of protein kinase Cgamma." Bioorg.Med.Chem.5(8). 1725-1737 (1997)

Kazuhiro Irie 等人：“蛋白激酶 Cgamma 的第一和第二富含半胱氨酸结构域的化学特征比较。”

Kazuhiro Irie et al.: "Synthesis and biological activities new conformationally restricted analogues of (-) -indolactam -V : elucidation of the biologically active conformation of the tumor-promoting teleocidins." J.Am.Chem.Soc.118(44). 10733-10743 (1996)

Kazuhiro Irie 等人：“(-)-indolactam -V 的新构象限制类似物的合成和生物活性：阐明促进肿瘤的 teleocidins 的生物活性构象。”

Kazuhiro Irie: "Function and role of the two zinc-finger motifs of protein kinase C." Kagaku-to-Seibutsu. 35(7). 638-639 (1997)

Kazuhiro Irie：“蛋白激酶 C 的两个锌指基序的功能和作用。”

Kazuhiro Irie: "Comparison of chemical characteristics of the first and the second cysteine-rich domains of protein kinase Cγ" Bioorg.Med.Chem.5・8. 1725-1737 (1997)

Kazuhiro Irie：“蛋白激酶 Cγ 的第一和第二富含半胱氨酸结构域的化学特性的比较”Bioorg.Med.Chem.5・8（1997）。