Joint study on design, synthesis and analysis of agonists and antagonists of galanin

甘丙肽是一种由29个氨基酸残基组成的神经肽，在脑和外周都具有多种生物活性。本论文在国际合作项目“甘丙肽激动剂拮抗剂的设计、合成与分析”中进行了以下研究：1）本实验室采用固相法或传统的溶液法合成了23个甘丙肽相关肽（N.Yanaihara）。2）人类的影响，比较了大鼠和猪甘丙肽对13.9mM葡萄糖刺激的胰岛素释放的影响，大鼠甘丙肽显示出对葡萄糖诱导的胰岛素释放的最高抑制。3）人和大鼠甘丙肽以及10 μ M的猪甘丙肽<-8>分别抑制GRP（14-27）刺激的胃泌素释放。4）人和大鼠甘丙肽以及猪甘丙肽在10 μ M浓度下<-6>均能抑制环肌的收缩反应。 ...更多信息 e以剂量依赖性方式。甘丙肽（1-15）-醇等甘丙肽类化合物对大肠杆菌的生长抑制作用几乎相等。这些结果强烈地表明甘丙肽抑制神经诱发的环行肌收缩的活性位点存在于该分子的氨基末端的一半。5）甘丙肽抑制隐神经制备物诱发的c-纤维反应0.01 μ M和更高浓度（1- 3 μ M）部分抑制单突触反射。人甘丙肽最强，猪甘丙肽最弱。氨基末端片段甘丙肽（1-15）-醇显示非常弱的作用。6）发现[D-Trp^&lt;8，9&gt;]-甘丙肽（1-15）-醇和[D-Thr^6，D-trp^&lt;8，9&gt;]-甘丙肽（1-15）-醇在体外对葡萄糖刺激的胰岛素释放起拮抗作用，而甘丙肽（1-13）和P物质（5-11）组成的肽在10 μ M时<-7>在该系统中不显示任何拮抗活性。特别是10 μ M的[D-Thr^6，D-Trp^&lt;8，9&gt;]-甘丙肽（1-15）-<-7>醇完全消除了10 μ <-9>M大鼠甘丙肽在离体灌流大鼠胰腺中的抑制作用。7）证明了特异性[^<125>I]甘丙肽（1-15）片段结合位点在大鼠脑中的存在和广泛分布。这些结合位点也存在于一些缺乏或具有很少的[^<125>I]甘丙肽（1-29）结合位点的区域，如背侧海马结构、新皮层和新纹状体。8）神经肽与P物质和其他速激肽的共存和相互作用，特别是甘丙肽。由于甘丙肽和相关肽在细胞或细胞外调节机制中的作用及其潜在的治疗价值的重要性，了解它们与特异性受体的相互作用，将为构效关系研究提供有用的信息。在这次国际联合研究中，对这一问题采取了各种各样的方法，11月在静冈举行的研讨会帮助取得了丰硕成果和成功。少

The neuropeptide galanin with 29 amino acid residues has been known to have numerous biological activities, both in the brain and in the periphery. The following studies have been carried out under International Joint study on Design, Synthesis and Analysis of Agonists Antagonists of Galanin.1) Twenty-three galanin-related peptides were synthesized by solid phase technology or conventional solution method in our laboratory (N.Yanaihara).2) Effects of human, rat and porcine galanins on 13.9mM glucose-stimulated insulin release were compared on glucose-induced insulin release in isolated rat pancreas.Among them, rat galanin showed the highest inhibition of glucose induced insulin release.3) Human and rat galanins as well as porcine galanin at 10^<-8>M inhibited GRP(14-27) stimulated gastrin release, respectively. Among them, rat galanin was the weakest in this system.4) Human and rat galanins as well as porcine galanin at 10^<-6>M suppressed the contractile response of the circular muscl … More e in a dose dependent manner, respectively. The galanins including galanin (1-15)-ol showed nearly equipotent inhibitory effect. These results suggest strongly the existence of the active site of galanin for its suppression of neurally-evoked circular muscle contractions in the amino-terminal half of the molecule.5) Galanins depressed the c-fiber response evoked by saphenous never preparation 0.01muM and higher concentrations (1-3muM) partially inhibited the monosynaptic reflex. Human galanin was the most potent and porcine galanin was the weakest. The amino-terminal fragment, galanin(1-15)-ol showed very weak effects.6) Both [D-Trp^<8,9>]-galanin (1-15)-ol and [D-Thr^6,D-trp^<8,9>]-galanin (1-15)-ol were found to act as antagonist on glucose-stimulated insulin release in vitro, while galantide, a peptide comprising galanin (1-13) and substance P(5-11), at 10^<-7>M did not show any antagonistic activity in this system. Especially, [D-Thr^6,D-Trp^<8,9>]-galanin (1-15)-ol at 10^<-7>M completely abolished the inhibitory effect of 10^<-9>M rat galanin in the isolated perfused rat pancreas.7) The existence and widespread distribution of specific [^<125>I] galanin (1-15) fragment binding sites in the rat brain was demonstrated. These binding sites were also present in several areas lacking or having very few [^<125>I] galanin (1-29) binding sites, such as the dorsal hippocampal formation, the neocortex and the neostriatum.8) Coexistence and interaction of neuropeptides with substance P and other tachykinins, with special reference to galanin were demonstrated.Because of the importance of galanin and the related peptides in the role of cellular or extracellular regulatory mechanisms and their potential therapeutic value, and understanding of their detailed interactions with the specific receptor would provide useful information for structure-activity study. A variety of approach to this problems have been performed in this international joint study, And Symposium help in November in Shizuoka was really fruitful and successful. Less