A synthetic dinuclear copper(II) hydrolase and its potential as antitumoral: cytotoxicity, cellular uptake, and DNA cleavage

J Inorg Biochem. 2009 Oct;103(10):1323-30. doi: 10.1016/j.jinorgbio.2009.05.008. Epub 2009 May 20.

Abstract

We have studied the protonation equilibria of a dicopper(II) complex [Cu(2)(micro-OH)(C(21)H(33)ON(6))](ClO(4))(2).H(2)O, (1), in aqueous solution, its interactions with DNA, its cytotoxic activity, and its uptake in tumoral cells. C(21)H(33)ON(6) corresponds to the ligand 4-methyl-2,6-bis[(6-methyl-1,4-diazepan-6-yl)iminomethyl]phenol. From spectrophotometric data the following pKa values were calculated 3.27, 4.80 and 6.10. Complex 1 effectively promotes the hydrolytic cleavage of double-strand plasmid DNA under anaerobic and aerobic conditions. The following kinetic parameters were calculated k(cat) of 2.73 x 10(-4)s(-1), K(M) of 1.36 x 10(-4)M and catalytic efficiency of 2.01 s(-1)M(-1), a 2.73 x 10(7) fold increase in the rate of the reaction compared to the uncatalyzed hydrolysis rate of DNA. Competition assays with distamycin reveal minor groove binding. Complex 1 inhibited the growth of two tumoral cell lines, GLC4 and K562, with the IC(50) values of 14.83 microM and 34.21 microM, respectively. There is a good correlation between cell growth inhibition and intracellular copper content. When treated with 1, cells accumulate approximately twice as much copper as with CuCl(2). Copper-DNA adducts are formed inside cells when they are exposed to the complex. In addition, at concentrations that compound 1 inhibits tumoral cell growth it does not affect macrophage viability. These results show that complex 1 has a good therapeutic prospect.

MeSH terms

  • Animals
  • Antineoplastic Agents / chemistry
  • Antineoplastic Agents / pharmacology*
  • Cell Survival / drug effects
  • Copper*
  • DNA Adducts / chemistry
  • DNA Adducts / metabolism
  • DNA Cleavage / drug effects*
  • Distamycins / pharmacology
  • Drug Screening Assays, Antitumor / methods
  • Female
  • Humans
  • K562 Cells
  • Kinetics
  • Macrophages, Peritoneal / metabolism
  • Male
  • Mice
  • Neoplasms / drug therapy*
  • Neoplasms / metabolism
  • Organometallic Compounds / chemistry
  • Organometallic Compounds / pharmacology*

Substances

  • Antineoplastic Agents
  • DNA Adducts
  • Distamycins
  • Organometallic Compounds
  • Copper
  • stallimycin