Zhang Luyao, Chen Chen, Zou Wanchen, Chen Xiaoling, Zhou Mei, Ma Chengbang, Xi Xinping, Chen Tianbao, Shaw Chris, Liu Mingchun, Wang Lei
Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, China.
School of Pharmacy, Queen's University Belfast, Belfast, United Kingdom.
Front Mol Biosci. 2022 Sep 29;9:953974. doi: 10.3389/fmolb.2022.953974. eCollection 2022.
Mammalian bombesin-like neuropeptides (BLPs) play an important role in regulation of physiological and pathophysiological processes. Frog skin-derived BLPs, of smaller size and diverse lengths and sequences at their N-terminus, have attracted the attention of many researchers. However, these N-terminal variants and the receptors modulating their pharmacological actions are poorly studied and less understood. In this study, two BLPs, namely, [Asn, Lys, Thr, Phe]3-14-bombesin and [Asn, Lys, Phe]3-14-bombesin with primary structures NLGKQWATGHFM and NLGKQWAVGHFM were isolated from the skin secretion of hybrid kl. . Both BLPs share a similar primary structure with only a single amino acid substitution at the eighth position (threonine to valine), while they have quite different myotropic potencies with EC values in the range of 22.64 ± 9.7 nM ( = 8) to 83.93 ± 46.9 nM ( = 8). The potency of [Asn, Lys, Thr, Phe]3-14-bombesin was approximately 3-fold higher than that of [Asn, Lys, Phe]3-14-bombesin. Through the investigation of receptor selectivity using a canonical bombesin receptor antagonist, it was found that [Asn, Lys, Thr, Phe]3-14-bombesin and [Asn, Lys, Phe]3-14-bombesin had an affinity to both BB1 and BB2 receptors. Their contractile functions are mainly modulated by both BB1 and BB2 receptors on rat urinary bladder and BB2 alone on rat uterus smooth muscle preparations. These data may provide new insights into the design of potent and selective ligands for bombesin receptors. Moreover, [Asn, Lys, Thr, Phe]3-14-bombesin and [Asn, Lys, Phe]3-14-bombesin did not induce significant hemolysis and toxicity in normal human cells, suggesting that these two natural novel BLPs have great potential for development into new drug candidates.
哺乳动物的蛙皮素样神经肽（BLPs）在生理和病理生理过程的调节中发挥着重要作用。来自蛙皮的BLPs，尺寸较小，N端长度和序列多样，吸引了众多研究人员的关注。然而，这些N端变体及其调节药理作用的受体的研究较少，了解也较少。在本研究中，从杂种蛙kl.的皮肤分泌物中分离出两种BLPs，即[天冬酰胺、赖氨酸、苏氨酸、苯丙氨酸]3 - 14 -蛙皮素和[天冬酰胺、赖氨酸、苯丙氨酸]3 - 14 -蛙皮素，其一级结构分别为NLGKQWATGHFM和NLGKQWAVGHFM。两种BLPs具有相似的一级结构，仅在第八位有一个氨基酸取代（苏氨酸被缬氨酸取代），而它们的肌动活性差异很大，EC值在22.64±9.7 nM（n = 8）至83.93±46.9 nM（n = 8）范围内。[天冬酰胺、赖氨酸、苏氨酸、苯丙氨酸]3 - 14 -蛙皮素的活性约比[天冬酰胺、赖氨酸、苯丙氨酸]3 - 14 -蛙皮素高3倍。通过使用经典的蛙皮素受体拮抗剂研究受体选择性，发现[天冬酰胺、赖氨酸、苏氨酸、苯丙氨酸]3 - 14 -蛙皮素和[天冬酰胺、赖氨酸、苯丙氨酸]3 - 14 -蛙皮素对BB1和BB2受体均有亲和力。它们的收缩功能主要由大鼠膀胱上的BB1和BB2受体以及大鼠子宫平滑肌制剂上的BB2受体单独调节。这些数据可能为设计强效和选择性的蛙皮素受体配体提供新的见解。此外，[天冬酰胺、赖氨酸、苏氨酸、苯丙氨酸]3 - 14 -蛙皮素和[天冬酰胺、赖氨酸、苯丙氨酸]3 - 14 -蛙皮素在正常人细胞中未诱导明显的溶血和毒性，表明这两种天然新型BLPs具有开发成新药候选物的巨大潜力。
