Elucidation of vasoactive intestinal peptide pharmacophore for VPAC1 receptors in human, rat, and guinea pig
Elucidation of vasoactive intestinal peptide pharmacophore for VPAC1 receptors in human, rat, and guinea pig
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DOI：
10.1124/jpet.301.1.37
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发表时间：
2002-04-01
影响因子：
3.5
通讯作者：
Jensen, RT
中科院分区：
文献类型：
--
作者：
Igarashi, H;Ito, T;Jensen, RT
血管活性肠肽（VIP）是一种神经递质，参与多种病理和生理过程。VIP快速降解，需要简化的稳定类似物。VIP的作用在大鼠和豚鼠中进行了广泛的研究。然而，它在这些物种中的药效团是否与人类相似在很大程度上是未知的。为了解决这个问题，我们通过使用丙氨酸和D-氨基酸扫描研究了VPAC（1）（癌症中的主要受体亚型，广泛分布于正常组织中）的VIP药效团。使用转染的中国仓鼠卵巢（CHO）和PANC 1细胞以及具有天然VPAC的细胞（1）评估了与大鼠、豚鼠和人VPAC（1）的相互作用。VIP药效团存在重要的种属差异。在CHO细胞中表达的人VPAC（1），在以前的研究中几乎完全使用，与T47 D细胞中的天然VPAC（1）明显不同。用于确定亲和力的最重要的氨基酸是His（1）、Asp（3）、Phe（6）、Arg（12）、Arg（14）和Leu（23）。Ser（2）、Asp（8）、Asn（9）、Thr（11）、瓦尔（19）、Asn（24）、Ser（25）、Leu（27）和Asn（28）对于高亲和力相互作用/活化不是必需的。合成了[Ala（2，8，9，11，19，24，25，27，28]）VIP，它含有11个丙氨酸，在所有物种中与VIP在VPAC（1）受体上的作用等效，代谢稳定。我们的研究结果表明，在VPAC（1）的简化VIP类似物的任何设计中，重要的是要考虑物种差异，并且必须使用反映天然受体药效团的转染系统。最后，我们的结果，一个简化的，代谢稳定的VIP类似物被确定，应该是有用的原型设计的选择性激动剂/拮抗剂，可能是有用的治疗。
Vasoactive intestinal peptide (VIP) is a neurotransmitter involved in a number of pathological and physiological processes. VIP is rapidly degraded and simplified stable analogs are needed. VIP's action was extensively studied in rat and guinea pig. However, it is largely unknown whether its pharmacophore in these species resembles human. To address this issue we investigated the VIP pharmacophore for VPAC(1) (the predominant receptor subtype in cancers and widely distributed in normal tissues) by using alanine and D-amino acid scanning. Interaction with rat, guinea pig, and human VPAC(1) was assessed using transfected Chinese hamster ovary (CHO) and PANC1 cells and cells possessing native VPAC(1). Important species differences existed in the VIP pharmacophore. The human VPAC(1) expressed in CHO cells, which were used almost exclusively in previous studies, differed markedly from the native VPAC(1) in T47D cells. The most important amino acids for determining affinity are His(1), Asp(3), Phe(6), Arg(12), Arg(14), and Leu(23). Ser(2), Asp(8), Asn(9), Thr(11), Val(19), Asn(24), Ser(25), Leu(27), and Asn(28) are not essential for high-affinity interaction/activation. [Ala(2,8,9,11,19,24,25,27,28])VIP, which contained 11 alanines, was synthesized and it was equipotent to VIP at VPAC(1) receptors in all species and was metabolically stable. Our results show in any design of simplified VIP analogs for VPAC(1) it will be important to consider species differences and it is essential to use transfected systems that reflect the native receptor's pharmacophore. Last, with our results a simplified, metabolically stable VIP analog was identified that should be useful as a prototype for design of selective agonists/antagonists that could be useful therapeutically.