Pająk Marek, Kamysz Elżbieta, Banach Marcin, Jankowski Wojciech Michał, Tarasiuk-Zawadzka Aleksandra, Fichna Jakub, Woźniczka Magdalena
Department of Physical and Biocoordination Chemistry, Medical University of Lodz, Muszyńskiego 1, 90-151, Lodz, Poland.
Laboratory of Chemistry of Biological Macromolecules, Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, 80-308, Gdansk, Poland.
Amino Acids. 2025 Sep 18;57(1):47. doi: 10.1007/s00726-025-03481-2.
The present work describes the protolytic properties and in vitro biological activity of spinorphin (Leu-Val-Val-Tyr-Pro-Trp-Thr) and three spinorphin derivatives containing butyric acid residue: Butyryl-Lys-Lys-Leu-Leu-Val-Tyr-Pro-Trp-Thr, Butyryl-Lys-Lys-Leu-Val-Val-Tyr-Pro-Trp-Thr (butyric acid bound to the α-amino group of lysine), Lys(Butyryl)-Lys-Leu-Val-Val-Tyr-Pro-Trp-Thr (butyric acid bound to the ε-amino group of lysine) in an aqueous solution. The overall protonation constants and the stepwise dissociation constants of the ligands studied were calculated by the potentiometric method. The percentage of each species formed was estimated from the species distribution curves as a function of pH. The biological activity of all tested compounds was characterized in vitro, in the neutral red uptake and Griess assay tests in RAW264.7 macrophage cell line. The three protonation constants for spinorphin and four for its derivatives suggest that metal ions may bind to these peptides and form complexes by coordination with the functional groups of the respective amino acid residues. In vitro biological activity tests suggest that two peptides deserve attention for their potential anti-inflammatory role.
本研究描述了斯皮诺啡（亮氨酸 - 缬氨酸 - 缬氨酸 - 酪氨酸 - 脯氨酸 - 色氨酸 - 苏氨酸）及其三种含有丁酸残基的斯皮诺啡衍生物：丁酰基 - 赖氨酸 - 赖氨酸 - 亮氨酸 - 亮氨酸 - 缬氨酸 - 酪氨酸 - 脯氨酸 - 色氨酸 - 苏氨酸、丁酰基 - 赖氨酸 - 赖氨酸 - 亮氨酸 - 缬氨酸 - 缬氨酸 - 酪氨酸 - 脯氨酸 - 色氨酸 - 苏氨酸（丁酸与赖氨酸的α - 氨基结合）、赖氨酸（丁酰基） - 赖氨酸 - 亮氨酸 - 缬氨酸 - 缬氨酸 - 酪氨酸 - 脯氨酸 - 色氨酸 - 苏氨酸（丁酸与赖氨酸的ε - 氨基结合）在水溶液中的质子解离性质和体外生物活性。通过电位滴定法计算所研究配体的总质子化常数和逐步解离常数。根据物种分布曲线估计每种物种形成的百分比与pH的函数关系。在RAW264.7巨噬细胞系的中性红摄取和格里斯试验中对所有测试化合物的生物活性进行了体外表征。斯皮诺啡的三个质子化常数及其衍生物的四个质子化常数表明金属离子可能与这些肽结合，并通过与各自氨基酸残基的官能团配位形成络合物。体外生物活性测试表明，两种肽因其潜在的抗炎作用值得关注。
