Synthetic Studies on Endothelin and Its Related Peptides Using a New S-Derotecting Reagent

新型S-去保护试剂合成内皮素及其相关肽的研究

• 批准号: 01571149
• 负责人: FUJII Nobutaka
• 金额: $ 1.34万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01571149/
• 关键词: Endothelin; Big Endothelin; S-Deprotecting Reagent; Silver Trifluoromethanesulphonate; トリメチルシリルブロマイド; 血管収縮作用; システインのSH保護基; Caイオンチャンネル

Endothelin (ET-1), a 21-residue peptide, characterized from aortic endothelial cells by Yanagisawa, et al., has a potent vasoconstrictor effect. Studies on cloning and sequencing of prepro ET cDNA have revealed the existence of the putative pecuraor of ET-21, named big endothelin (big ET), consisting of 38 amino acid residues. In addition, other types of ET were found and named ET-II, and ET-III. Big ET could be further processed to ET-21 by a unique endopeptidase named "enodothelin converting enzyme" (ECE). In order to evaluate the physiological significance of ET and clarify the chemical nature of ECE, we have synthesized ETs (ETI,II,III,) human big ET and their related peptides by two alternative solid phase method. In the course of this synthetic studies, a new Sdeprotecting reagent, silver trifluoromethanesulphonate, and a new two step hard acid deprotection/cleavage procedure [1._3SiBr deprotection and 2. HF(or Me_3SiOSO_2CF_3) cleavage] were developed.Boc-based solid phase synth … More esis of ET-1 was carried out by using automatic synthesizer. As amino acid derivatives, Cys(Acm), Asp(OBzl), Glu(OBzl), Ser(Bzl), Thr(Bzl), Trp(CHO), His(Tos), Tyr(BrZ), Lys(CIZ), and Arg(Mts) were employed. In the final step, the above two new methods were successfully employed to give a biologically active ET-I. In essentially the same manner, human big endothelin (ET-38) and C-terminally elongated homologs of ET-I, such as ET-22, ET-23, ET-31, and ET-36 were prepared. ET-II and ET-III were synthesized manually by Fmoc-based solid phase synthesis according to the principle of Sheppard, et al., using S-p-methoxybenzyl (MBzl) protection for 4 Cys residues. ETs, big ET, and their related peptides thus synthesized were assessed to the several bioassay systems.The in vitro vasoconstrictor activity of human big ET was two orders of magnitude less than that of ET-I. However, in vivo assay in rats, human big ET caused a dose dependent rise of arterial pressure after the cumulative administration in essentially the same potency as that of ET-I. The activities of C-terminally elongated peptides, ET-22, ET-23, ET-31, and ET-36, were comparable to that human big ET. Thus, it was suggested that big-ET and all of C-terminally elongated peptides might be processed directly to ET-21 by ECE in animal body. Less

内皮素 (ET-1) 是一种 21 个残基的肽，由 Yanagisawa 等人从主动脉内皮细胞中鉴定，具有有效的血管收缩作用。对前 ET cDNA 的克隆和测序研究揭示了 ET 假定特有的存在。 -21，命名为大内皮素（big ET），由38个氨基酸残基组成。此外，还发现了其他类型的ET并命名。 ET-II和ET-III可以通过一种独特的肽链内切酶“内皮素转化酶”(ECE)进一步加工成ET-21，以评估ET的生理意义并阐明ECE的化学性质。我们通过两种替代固相方法合成了 ET（ETI、II、III）人类大 ET 及其相关肽。在本合成研究过程中，我们使用了一种新的 S 脱保护试剂银。三氟甲磺酸盐，并开发了一种新的两步硬酸脱保护/裂解程序[1._3SiBr脱保护和2. HF(或Me_3SiOSO_2CF_3)裂解]。基于Boc的固相合成…更多ET-1的esis是通过使用自动进行的作为氨基酸衍生物，Cys(Acm)、Asp(OBzl)、Glu(OBzl)、最后一步采用了Ser(Bzl)、Thr(Bzl)、Trp(CHO)、His(Tos)、Tyr(BrZ)、Lys(CIZ)和Arg(Mts)，成功地实现了上述两种新方法。用于以基本相同的方式产生生物活性ET-1，人大内皮素(ET-38)和ET-1的C末端延长的同源物，例如ET-22，根据Sheppard等人的原理，使用S-对甲氧基苯甲基(MBzl)，通过基于Fmoc的固相合成手动合成ET-23、ET-31和ET-36。通过几种生物测定系统评估了由此合成的4个Cys残基的保护作用。体外血管收缩活性。人大ET比ET-I低两个数量级，然而，在大鼠体内测定中，人大ET在累积给药后引起动脉压的剂量依赖性升高，其效力与ET-I基本相同。 C 末端延长的肽 ET-22、ET-23、ET-31 和 ET-36 的活性与人类大 ET 相当，因此，建议大 ET 和所有肽。 C 末端延长的肽可能在动物体内被 ECE 直接加工成 ET-21。

项目成果

Nobutaka Fujii他: "Silver Trifluoromethanesulfonate as an S-Deprotecting Reagent for the Synthesis of Cystine Peptide" J.Chem.Soc.,Chem.Commun.283-284 (1989)

Nobutaka Fujii 等人：“三氟甲磺酸银作为胱氨酸肽合成的 S-去保护试剂”J.Chem.Soc.，Chem.Commun.283-284 (1989)

Motoyoshi Nomizu, Yoshimasa Iagaki, Takeyoshi Yamashita, Ako Okubo, Akira Otaka, Nobutaka Fujii, Peter P. Roller, and Haruaki Yajima: "Two-Step Hard Acid Deprotection/Cleavage Procedure for Solid Phase Peptide Synthesis" Int. J. Peptide Protein Res.

Motoyoshi Nomizu、Yoshimasas Iagaki、Takeyoshi Yamashita、Ako Okubo、Akira Otaka、Nobutaka Fujii、Peter P. Roller 和 Haruaki Yajima：“固相肽合成的两步硬酸脱保护/裂解程序” Int。

Motoyoshi Nomizu: "Solid Phase Peptide Synthesis of Human Endothelin Precursor Peptides Using TwoーStep Hard Acid Deprotection/Cleavage Methods;" Int J.Peptide Protein Res.

Motoyoshi Nomizu：“使用两步硬酸脱保护/切割方法固相合成人内皮素前体肽；”Int J.Peptide Protein Res。

Hiroshi Morimoto: "Silver Trifluoromethanesulphonate,as an SーAcm Deprotecting Reagent and its Application to The Synthesis of Endothelin and EndothelinーLike Peptide" Peotide Chemistry. 211-214 (1989)

Hiroshi Morimoto：“三氟甲磺酸银，作为 S-Acm 脱保护试剂及其在内皮素和内皮素样肽合成中的应用”Peotide Chemistry 211-214 (1989)。

Nobutaka Fujii: "Silver Trifluoromethanesulphonate as an SーDeprotecting Reagent for the Synthesis of Cystine Peptides" J.Chem.Soc.,Chem.Commun.283-284 (1989)

Nobutaka Fujii：“三氟甲磺酸银作为胱氨酸肽合成的 S-去保护试剂”J.Chem.Soc.,Chem.Commun.283-284 (1989)