Mapping the active site in vasoactive intestinal peptide to a core of four amino acids: Neuroprotective drug design
Mapping the active site in vasoactive intestinal peptide to a core of four amino acids: Neuroprotective drug design
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DOI：
10.1073/pnas.96.7.4143
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发表时间：
1999-03-30
影响因子：
11.1
通讯作者：
Fridkin, M
中科院分区：
文献类型：
--
作者：
Gozes, I;Perl, O;Fridkin, M
了解导致肽作用的分子机制需要鉴定核心活性位点。主要的28-aa神经肽，血管活性肠肽（VIP），提供神经保护。在N-末端具有硬脂基部分和正亮氨酸残基取代Met-17的亲脂性衍生物在促进神经元存活方面比VIP有效100倍，作用于飞摩尔-皮摩尔浓度。为了确定VIP中的活性位点，设计、合成了50多个含有N-末端硬脂酸连接和酰胺化C末端的相关片段，并测试了其神经保护特性。Stearyl-Lys-Lys-Tyr-Leu-NH 2（衍生自VIP和相关肽垂体腺苷酸环化酶激活肽的C末端）捕获由整个28-aa母体亲脂性衍生物提供的神经营养作用，并在体外保护免受β-淀粉样蛋白毒性。此外，4-aa亲脂肽识别VIP结合位点，增强胆碱乙酰转移酶活性以及阿尔茨海默病相关的体内模型中的认知功能。鼻内给药放射性标记的肽后的生物分布研究表明，给药后30分钟脑中的肽是完整的。因此，亲脂性肽片段提供生物利用度和稳定性，为针对神经退行性疾病的药物设计提供先导化合物。
The understanding of the molecular mechanisms leading to peptide action entails the identification of a core active site. The major 28-aa neuropeptide, vasoactive intestinal peptide (VIP), provides neuroprotection. A lipophilic derivative with a stearyl moiety at the N-terminal and norleucine residue replacing the Met-17 was 100-fold more potent than VIP in promoting neuronal survival, acting at femtomolar-picomolar concentration. To identify the active site in VIP, over 50 related fragments containing an N-terminal stearic acid attachment and an amidated C terminus were designed, synthesized, and tested for neuroprotective properties. Stearyl-Lys-Lys-Tyr-Leu-NH2 (derived from the C terminus of VIP and the related peptide, pituitary adenylate cyclase activating peptide) captured the neurotrophic effects offered by the entire 28-aa parent lipophilic derivative and protected against beta-amyloid toxicity in vitro. Furthermore, the 4-aa lipophilic peptide recognized VIP-binding sites and enhanced choline acetyltransferase activity as well as cognitive functions in Alzheimer's disease-related in vivo models. Bio distribution studies following intranasal administration of radiolabeled peptide demonstrated intact peptide in the brain 30 min after administration. Thus, lipophilic peptide fragments offer bioavailability and stability, providing lead compounds for drug design against neurodegenerative diseases.