Laboratory of Chemistry, Department of Biosciences, University of Kuopio, Kuopio, Finland.
Amino Acids. 2010 Feb;38(2):501-7. doi: 10.1007/s00726-009-0409-6. Epub 2009 Dec 2.
Ionic interactions are essential for the biological functions of the polyamines spermidine and spermine in mammalian physiology. Here, we describe a simple gram scale method to prepare 1,12-diamino-3,6,9-triazadodecane (SpmTrien), an isosteric charge-deficient spermine analogue. The protonation sites of SpmTrien were determined at pH range of 2.2-11.0 using two-dimensional (1)H-(15)N NMR spectroscopy, which proved to be more feasible than conventional methods. The macroscopic pK(a) values of SpmTrien (3.3, 6.3, 8.5, 9.5 and 10.3) are significantly lower than those of 1,12-diamino-4,9-diazadodecane (spermine). The effects of SpmTrien and its parent molecule, 1,8-diamino-3,6-diazaoctane (Trien), on cell growth and polyamine metabolism were investigated in DU145 prostate carcinoma cells. SpmTrien downregulated the biosynthetic enzymes ornithine decarboxylase (ODC) and S-adenosyl-L: -methionine decarboxylase and decreased intracellular polyamine levels, whereas the effects of Trien alone were minor. Interestingly, both SpmTrien and Trien were able to partially overcome growth arrest induced by an ODC inhibitor, alpha-difluoromethylornithine, indicating that they are able to mimic some functions of the natural polyamines. Thus, SpmTrien is a novel tool to influence polyamine interaction sites at the molecular level and offers a new means to study the contribution of the protonation of spermine amino group(s) in the regulation of polyamine-dependent biological processes.
离子相互作用对于多胺亚精胺和精胺在哺乳动物生理学中的生物学功能至关重要。在这里，我们描述了一种简单的克级规模方法来制备 1,12-二氨基-3,6,9-三氮杂十二烷（SpmTrien），这是一种等电子电荷缺失的精胺类似物。使用二维（1）H-（15）N NMR 光谱法在 pH 值为 2.2-11.0 的范围内确定了 SpmTrien 的质子化位点，这比传统方法更可行。SpmTrien 的宏观 pK（a）值（3.3、6.3、8.5、9.5 和 10.3）明显低于 1,12-二氨基-4,9-二氮杂十二烷（精胺）的 pK（a）值。在 DU145 前列腺癌细胞中研究了 SpmTrien 和其母体分子 1,8-二氨基-3,6-二氮杂辛烷（Trien）对细胞生长和多胺代谢的影响。SpmTrien 下调了生物合成酶鸟氨酸脱羧酶（ODC）和 S-腺苷-L：-甲硫氨酸脱羧酶，并降低了细胞内多胺水平，而单独使用 Trien 的作用较小。有趣的是，SpmTrien 和 Trien 都能够部分克服 ODC 抑制剂 α-二氟甲基鸟氨酸诱导的生长停滞，表明它们能够模拟天然多胺的某些功能。因此，SpmTrien 是一种影响多胺相互作用位点的新型工具，可以在分子水平上影响多胺相互作用位点，并提供一种新的方法来研究精胺氨基（s）质子化在调节多胺依赖性生物过程中的贡献。
