Szczepanik Wojciech, Czarny Anna, Zaczyńska Ewa, Jezowska-Bojczuk Małgorzata
Faculty of Chemistry, University of Wroclz.shtsls;aw, 14 F. Joliot-Curie, 50-383, Wrocław, Poland.
J Inorg Biochem. 2004 Feb;98(2):245-53. doi: 10.1016/j.jinorgbio.2003.10.013.
The widespread presence of pathogenic bacteria is a cause of permanent demand for investigating the properties of antimicrobial agents. The chemical basis of several toxic effects induced by antibiotics still remains unclear. Aminoglycosides, highly ototoxic and nephrotoxic drugs, are capable of copper(II) ions chelating. In this study we established the affinity of kanamycin A towards copper(II), in contrast with other metal ions: iron(III), nickel(II), cobalt(II) and zinc(II) by means of potentiometry. Circular dichroism spectroscopy was applied to monitor the competition of copper(II) partition between kanamycin A and human serum albumin. We show, that the drug is able to digest Cu(II) ions from HSA to some extent and comparing the stability constants for metal and antibiotic with those, obtained for the N-terminal Asp-Ala-His-Lys (DAHK) sequence, which constitutes a copper(II) binding domain within albumin, we demonstrate that the Cu(II)-kanamycin A complex formation is possible also in blood plasma. Bioassays and immunoassay were used to find out the possibility of Cu(II)-kanamycin A complexes to induce cytokines: tumor necrosis factor (TNF), interferon (IFN) and interleukin-10 (IL-10) in human peripheral blood leukocytes. The effect on the cytokines release was dose and time dependent and the interdependence between IL-10 and TNF stimulation was found. We report that Cu(II)-aminoglycoside systems can act as moderate inducers of TNF-alpha, IFN-alpha/beta and IL-10 released from human leukocytes. We have also found that these complexes are non-toxic for human A549 cells.
致病细菌的广泛存在是对抗菌剂特性进行研究的永久需求的一个原因。抗生素诱导的几种毒性作用的化学基础仍不清楚。氨基糖苷类药物具有高度耳毒性和肾毒性，能够螯合铜（II）离子。在本研究中，我们通过电位滴定法确定了卡那霉素A与铜（II）的亲和力，与其他金属离子：铁（III）、镍（II）、钴（II）和锌（II）形成对比。应用圆二色光谱法监测卡那霉素A和人血清白蛋白之间铜（II）分配的竞争。我们表明，该药物能够在一定程度上从人血清白蛋白中消化铜（II）离子，并且通过比较金属与抗生素的稳定常数以及与N端天冬氨酸-丙氨酸-组氨酸-赖氨酸（DAHK）序列（该序列构成白蛋白内的铜（II）结合结构域）所获得的稳定常数，我们证明在血浆中也可能形成铜（II）-卡那霉素A复合物。使用生物测定法和免疫测定法来研究铜（II）-卡那霉素A复合物在人外周血白细胞中诱导细胞因子：肿瘤坏死因子（TNF）、干扰素（IFN）和白细胞介素-10（IL-10）的可能性。对细胞因子释放的影响具有剂量和时间依赖性，并且发现了IL-10和TNF刺激之间的相互依赖性。我们报告铜（II）-氨基糖苷类系统可作为从人白细胞中释放的TNF-α、IFN-α/β和IL-10的中度诱导剂。我们还发现这些复合物对人A549细胞无毒。
