Development of highly potent vasoactive compounds.

开发高效血管活性化合物。

• 批准号: 11694281
• 负责人: NAKATA Yoshihiro
• 金额: $ 2.75万
• 依托单位: Hiroshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694281/
• 关键词: Maxadilan; sand fly; PACAP; PAC1 receptor; α helix; VIP; cAMP; αヘリックス構造; ジスルフィルド結合; PACAP1型受容体; 細胞内cAMP; melanophore培養細胞

Maxadilan (Maxa) is a potent and persistent vasodilator, which has been isolated from the salivary gland lysates of the blood feeding sand fly Lutzomyia longipalpis (a vector of leishmaniasis) by Lerner. Maxa consists of 61 amino acids with two disulfide linages. More recently, Maxa was found to be an agonist of the PACAP type 1 receptor (PAC1) although there is no significant sequence similarity between PACAPs and Maxa. PACAPs belong to a large family of neuropeptides that function through members spanning G-protein-coupled receptors. Exploring the recognition characteristics of GPCRs is particular important for the elucidation of their functions and development of pharmaceuticals. To develop agonists and antagonists against PAC1 , Maxa with its disulfide isomers and various related poptides such as middle, N- and C-terminal fragments as well as middle-region deleted Maxa have been prepared by highly efficient SPPS with improved synthesis protocols, After purification and characterization the peptides obtained were used in different bioassays that include a PAC 1 binding assay using rat brain membranes and the recent developed melanophore technology to elucidate the structure requirements for PAC1 recognition, especially anatgonistic actions. The results indicated that the middle section of Maxa and the first disulfide linkage are not essential for recognition. An antagonistic action to PAC1 is observed for the middle-region deleted Maxa fragment.

Maxadilan （Maxa）是一种强效持久的血管扩张剂，由Lerner从吸血沙蝇长掌Lutzomyia longipalpis（利什曼病的一种媒介）的唾液腺裂解物中分离得到。Maxa由61个氨基酸和两个二硫链组成。最近，Maxa被发现是PACAP 1型受体（PAC1）的激动剂，尽管PACAPs和Maxa之间没有明显的序列相似性。pacap属于一个神经肽大家族，通过跨越g蛋白偶联受体的成员发挥作用。探索gpcr的识别特性对阐明其功能和开发药物具有重要意义。为了开发针对PAC1的激动剂和拮抗剂，利用高效的SPPS技术，改进了合成方案，制备了具有二硫异构体的Maxa及其相关肽，如中间、N端和c端片段以及中间区域缺失的Maxa。纯化和鉴定后获得的肽用于不同的生物测定，包括使用大鼠脑膜的PAC1结合试验和最近开发的黑素细胞技术来阐明PAC1识别的结构要求，特别是拮抗作用。结果表明，Maxa的中段和第一二硫键不是识别的必要条件。在中部缺失的Maxa片段中观察到对PAC1的拮抗作用。

项目成果

K.Nokihara,T.Yasuhara,Y.Nakata,V.Wray and E.Lerner: "Synthesis and binding studies of Maxadilan, which is isolated from salivary gland of a sand fly, and its releated peptide on PACAP type 1 receptors"Peptide Science 1999, N. Fujii(Ed.). 29-32 (2000)

K.Nokihara、T.Yasuhara、Y.Nakata、V.Wray 和 E.Lerner：“从白蛉唾液腺中分离出的 Maxadilan 的合成和结合研究，及其在 PACAP 1 型受体上的相关肽”肽

K.Nokihara,T.Yasuhara,Y.Nakata,V.Wray and E.Lerner: "Synthesis and binding studies of Maxadilan, which is isolated from salivary gland of a sand fly, and its related peptide on PACAP type 1 receptors"Peptide Science 1999, N.Fujii (Ed.). 29-32 (2000)

K.Nokihara、T.Yasuhara、Y.Nakata、V.Wray 和 E.Lerner：“从沙蝇唾液腺中分离出的 Maxadilan 及其对 PACAP 1 型受体的相关肽的合成和结合研究”肽

Kiyoshi Nokihara, Tadashi Yasuhara, Yoshihiro Nakata, N.Fujii (Ed.): "Victor Wray and Ethan Lerner Synthesis and binding studies of Maxadian, which is isolated Salivery gland of a sand fly, and its related peptide on PACAP type 1 receptors."Peptide Scienc

Kiyoshi Nokihara、Tadashi Yasuhara、Yoshihiro Nakata、N.Fujii（主编）：“Victor Wray 和 Ethan Lerner 对 Maxadian 的合成和结合研究，Maxadian 是白蛉的分离唾液腺，及其在 PACAP 1 型受体上的相关肽。