Development of efficient synthetic methods for peptides and syntheses of self-defense peptides possessing anti-virus activity

高效肽合成方法的开发及具有抗病毒活性的自卫肽的合成

• 批准号: 05671748
• 负责人: OTAKA Akira
• 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 General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671748/
• 关键词: Solid-phase synthesis; Disulfide bond; Self-defense peptide; Protegrin; Defensin; Avidin-biotin; Affinity purification

Development of the solid-phase techniques for peptide synthesis have facilitated the syntheses of peptides, especially small and less complicated peptides. However, syntheses of large and/or multi disulfide containing-peptides have been encountered with some dufficulties, one being difficulties of final product purification due to the contamination of a number of deletion peptides and the other difficulties of controling egioselectivity of multi-disulfide bond formation. In order to circumvent these problems, we have developed new synthetic methodologies involving a affinity-purification method compatible with solid-phase peptide synthesis and disulfide bond-forming methods for peptides difficult to be solved in aqueous solvents which were utilized generally for disulfide bond formation.1.Development of the affinity-purification method utilizing the avidin-biotin system have been achieved. This methodology would facilitate the purification of peptides synthesized by solid-phase techniques.2.New cysteine S-protecting group (2-quinolinylmethyl) have been developed. The peptide bearing this protecting groups can be converted to the corresponding cystine peptide in DMF without solubility problem.3.We have investigated the feasibility of DMSO-mediated disulfide bond-forming reaction by formation of two-disulfide bond of apamin. And the new disulfide bond-forming reaction using AgOTf-DMSO/HCI system have been developed.4.Based on the above findings, we have achieved the syntheses of self-defense peptides [protegrins (18 aa, 2 S-S) and defensins (29-34 aa, 3 S-S) ] .

固相合成多肽技术的发展促进了多肽的合成，特别是小而不复杂的多肽的合成。然而，合成大型和/或多含二硫肽遇到了一些困难，一是由于许多缺失肽的污染导致最终产物纯化困难，二是控制多二硫键形成的地理选择性困难。为了避免这些问题，我们开发了新的合成方法，包括一种适用于固相肽合成的亲和纯化方法，以及通常用于二硫键形成的难以在水溶液中溶解的肽的二硫键形成方法。利用亲和素-生物素系统的亲和纯化方法已经得到了发展。该方法为固相合成多肽的纯化提供了便利。开发了新的半胱氨酸s保护基团（2-喹啉基甲基）。具有这些保护基团的肽可以在DMF中转化为相应的胱氨酸肽而没有溶解度问题。我们研究了二甲基亚砜介导的二硫键形成反应的可行性。利用AgOTf-DMSO/HCI体系建立了新的二硫键形成反应。基于以上发现，我们实现了自卫肽[蛋白蛋白（18 aa, 2 S-S）和防御素（29-34 aa, 3 S-S）]的合成。

项目成果

H.Tamamura et al.: "Disulfide bond-forming reaction using dimethylsulfoxide/aqueous HCI system and its application to regioselective two disulfide bond formation" Int.J.Peptide Protein Res.45 (in press). (1995)

H.Tamamura 等人：“使用二甲亚砜/HCl 水溶液系统的二硫键形成反应及其在区域选择性两个二硫键形成中的应用”Int.J.Peptide Protein Res.45（出版中）。

H.Nakashima et al.: "Defensins inhibit HIV replication in vitro" AIDS. 7. 1129-1129 (1993)

H.Nakashima 等人：“防御素在体外抑制 HIV 复制”艾滋病。

H.Yoshizawa et al.: "Direct disulfide formation from 2-quinolinylmethyl thioethers with iron (III) or copper (II) salt" Chemistry Letters. 803-806 (1993)

H.Yoshizawa 等人：“2-喹啉基甲基硫醚与铁 (III) 或铜 (II) 盐直接形成二硫化物”化学快报。

H.Yoshizawa et al.: "Direct disulfide formation from 2-quinolinylmethyl thioethers with iron(III)or copper(II)salt" Chemistry Letters. 803-806 (1993)

H.Yoshizawa 等人：“2-喹啉基甲基硫醚与铁（III）或铜（II）盐直接形成二硫化物”化学快报。

H.Tamamura et al.: "Disulfide bond formation in S-acetamidomethyl cysteine-containing peptides by the combination of silver trifluoromethanesulfonate and dimethylsulfoxide/aqueous HCI" Tetrahedron Letters. 34. 4931-4934 (1993)

H.Tamamura 等人：“通过三氟甲磺酸银和二甲基亚砜/HCl 水溶液的组合在含 S-乙酰氨基甲基半胱氨酸的肽中形成二硫键”四面体快报。