Superoxide dismutase mimetic activity of cytokinin-copper(II) complexes

Dissociation constants of cytokinins, derivatives of purine which form complexes with cupric ion, were determined by spectrophotometry and the stability constants of their copper complexes by pH titration. The values found for kinetin were 3.76, 9.96, 7.8, and 15.3 for pK1, pK2, logk1, and log beta 2, respectively, and those for 6-benzylaminopurine were, in the same order, 3.90, 9.84, 8.3, and 15.9. The copper(II) complexes with kinetin and 6-benzylaminopurine had superoxide dismutase mimetic activity, and the reaction rate constants with superoxide, which were determined by polarography, were 2.3 X 10(-7) M-1 s-1 for kinetin and 1.5 X 10(-7) M-1 s-1 for 6-benzylaminopurine at pH 9.8 and 25 degrees C.     

Potentiometric measurement of stability constants of complexes between fulvic acid carboxylate and Fe3+

The stability constants of complexes formed between iron (III) and fulvic acid extracted from organic manures and wastes such as urban domestic sewage sludge, farmyard manure, poultry manure and sulfitation pressmud were investigated by the potentiometric titration method in an ionic medium of 0.1 M KNO₃ at 25±1 °C. A modification of the Katchalsky's model was employed for the estimation of stability constants. The displacement of the titration curves due to presence of Fe³⁺ in FA solutions formed the basis of calculations. The weak acidic property of fulvic acids due to carboxyl groups resulted in buffering over a wide range of pH; fulvic acids were completely neutralized in the pH range of 7.00–8.85. Apparent dissociation constants (pK_APP) of weakly acidic carboxyl groups were a direct function of degree of dissociation (α_L) in the mid-range of titration curves but were non-linear at high and low α_L values. The stability constants for formation of Fe–FA complexes (log β_Fe) calculated from the titration data were in the range of 5.64–7.55, depending upon α_L and electrostatic properties of fulvic acids. The relatively high stability constants of Fe–FA complexes in comparison to those with other competing cations suggest that the Fe–FA complexes are relatively stable in a soil environment. This revised version was published online in June 2006 with corrections to the Cover Date.     

Copper(II) interaction with 3,5-diisopropylsalicylic acid (Dips): new insights on its role as a potential *OH inactivating ligand

The purpose of this study was to identify the low molecular mass complexes formed between copper(II) and 3,5-diisopropylsalicylic acid (Dips) in physiological conditions. Copper(II)-Dips complex equilibria were determined using glass electrode potentiometry and their solution structures checked by UV-visible (UV-vis) spectrophotometry. Because of the low solubility of Dips in water, the equilibria were investigated in different water/ethanol mixtures. Formation constants were extrapolated to 100% water and then compared with the values obtained for the other anti-inflammatory drugs previously studied. Given the prime role of histidine as the copper(II) ligand in blood plasma, copper(II)-histidine-Dips ternary equilibria were studied under similar experimental conditions. Computer simulations of copper(II) distribution relative to different biofluids, gastrointestinal (g.i.) fluid and blood plasma, show that like salicylic and anthranilic acids, Dips favors g.i. copper absorption, but cannot exert any significant influence on plasma copper distribution. Moreover, Dips can mobilize increasing fractions of copper(II) as the pH decreases. In conclusion, Dips seems to correspond to the notion of *OH-inactivating ligand (OIL) as determined for anthranilic acid.     

Speciation of copper(II) in natural waters in the presence of ligands of high and intermediate strength

Abstract

A new procedure is presented for the determination of the ligands of copper(II) in natural waters, based on titration with the metal ion, monitored by measuring the concentration of copper(II) sorbed on the carboxylic resin Amberlite CG 50. The data are treated by the Ruzic linearization method to obtain the concentration of the ligands and the conditional stability constant of the complexes. Ligands with reaction coefficient α_M higher than 0.1 K*w/V are detected, where K* is the ratio of the concentration of sorbed metal to the concentration of free metal in solution, which can be evaluated from the sorption equilibria of copper(II) on Amberlite CG 50, w is the amount of water in the resin phase, and V the volume of the solution phase. Some natural waters at high and low salinity were examined. The ligand concentration determined in these samples ranged from around 50 to 2000 nM, while the original copper concentrations from 11 to 130 nM. The ligand concentration was always much higher than that of copper(II). The conditional stability constants were very high, particularly in low salinity waters, where values as high as K’= 10^15.7 were obtained. In high salinity waters values around 10⁹ were found for the complex formation constant of the ligands titrated with copper(II). The investigation was also extended to a model solution, containing EDTA, obtaining K’ = 10^15.5, in acceptable agreement with that evaluated from the literature values.     

Characteristics of the mechanism of action of complex copper (II) compounds with alpha-amino acids

There was no direct inhibition of DNA synthesis in ascites hepatoma 22A cells after intraperitoneal injection of single doses of copper (II) complexes with amino acids into tumor-bearing C3HA mice. Meanwhile cis-dichlorodiamine platinum (II) (DDP) as well as sarcolysine showed such inhibition. Copper (II) complexes with alpha-amino acids displayed as significant superoxide dismutase-like activity at concentrations corresponding to therapeutic doses of these compounds. The complexes of copper (II) combined with DDP give an additive antitumor effect in solid tumors of mice.     

The interaction of copper(II) and glycyl-L-histidyl-L-lysine, a growth-modulating tripeptide from plasma.

The interaction between Cu(II) and the growth-modulating tripeptide glycyl-L-histidyl-L-lysine in the presence and absence of L-histidine was investigated by potentiometric titration and visible-absorption spectrophotometry at 25 degrees C in 0.15 M-NaCl. Analyses of the results in the pH range 3.5--10.6 indicated the presence of multiple species in solution in the binary system and extensive amounts of the ternary complexes in the ternary system. The species distribution and the stability constants, as well as the visible-absorption spectra of the species, were evaluated. The combined results were used to propose the structure of some of the complexes. The influence of the epsilon-amino group of the peptide in the enhancement of the stability constants was reflected prominently when compared with those complexes formed by either glycyl-L-histidine or glycyl-L-histidylglycine. The results obtained from the equilibrium-dialysis experiments showed that this tripeptide was able to compete with albumin for Cu(II) at pH 7.5 and 6 degrees C. At equimolar concentrations of albumin and the peptide, about 42% of the Cu(II) was bound to the peptide. At the physiologically relevant concentrations of Cu(II), albumin, L-histidine and this peptide, about 6% of the Cu(II) was associated with the low-molecular-weight components. This distribution could be due to the binary as well as the ternary complexes. The possible physiological role of these complexes in the transportation of Cu(II) from blood to tissues is discussed.     

Analytical determination of apparent stability constants using a copper ion selective electrode

Copper(II) complexes of di-, tri- and tetra peptides with previously published protonation constants were re-investigated using pH and copper ion selective electrode (ISE) potentiometry in conjunction with a modified version of HYPERQUAD computer program. The purpose was to demonstrate the suitability of the ISE approach for the determination of apparent stability constants for copper(II) complexes with ligands for which proton stability constants were not available. The interactions of Cu²⁺ with oligopeptides were also analysed using surface enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-ToF-MS). The results provide an insight into the metal complex species formed, their apparent stabilities under selected conditions and the effect of the relative positions of certain amino acids within the peptide sequence.     

Metal complexes of salicylhydroxamic acid (H2Sha), anthranilic hydroxamic acid and benzohydroxamic acid. Crystal and molecular structure of [Cu(phen)2(Cl)]Cl x H2Sha, a model for a peroxidase-inhibitor complex

Stability constants of iron(III), copper(II), nickel(II) and zinc(II) complexes of salicylhydroxamic acid (H2Sha), anthranilic hydroxamic acid (HAha) and benzohydroxamic acid (HBha) have been determined at 25.0 degrees C, I=0.2 mol dm(-3) KCl in aqueous solution. The complex stability order, iron(III) > copper(II) > nickel(II) approximately = zinc(II) was observed whilst complexes of H2Sha were found to be more stable than those of the other two ligands. In the preparation of ternary metal ion complexes of these ligands and 1,10-phenanthroline (phen) the crystalline complex [Cu(phen)2(Cl)]Cl x H2Sha was obtained and its crystal structure determined. This complex is a model for hydroxamate-peroxidase inhibitor interactions.     

Stability constants of thiocyanato complexes of cobalt(II), nickel(II) and copper(II) in methanol.

A spectrophotometric study of cobalt(II), nickel(II) and copper(II) thiocyanato complexes was carried out in methanol at 25 degrees C and at a constant ionic strength of 1 M. Under the experimental conditions, two mononuclear complexes are identified with each of the three metal ions. Their stability constants are determined with a recent PC program SIRKO and the calculated values are: for cobalt, log beta 1 = 1.6, log beta 2 = 2.7; for nickel, log beta 1 = 1.8, log beta 2 = 3.0; and for copper, log beta 1 = 3.0, log beta 2 = 3.6.     

Copper(II) and nickel(II) ternary complexes of L-dopa and related compounds

The stability constants of the mixed ligand complexes of L-dopa, L-tyrosine, L-phenylalanine, and dopamine with copper(II) and nickel(II) ions and with 2,2'-bipyridyl and 1,10-phenanthroline were determined pH-metrically at 25 degrees C and an ionic strength of 0.2 mol/dm3 (KCl). Spectral studies were made to establish the binding mode of the ambidentate L-dopa in the ternary complexes. In contrast with the aromatic (N,N) donor atoms, the (O,O) binding mode of L-dopa is particularly favored in its ternary systems with copper(II) and nickel(II); thus, even at physiological pH there is a very considerable formation of (O,O)-bound mixed ligand complexes containing a free amino acid side-chain. Numerous binary transition metal-L-dopa complexes and the ternary complexes formed with various B ligands have been evaluated from a coordination chemistry aspect, with regard to the possibility of their therapeutic application in the treatment of Parkinson disease.     

Complexes of aminophosphonates. I. Transition metal complexes of aminophosphonic acid analogues of α-alanine, β-alanine,phenylalanine and tyrosine

The stoichiometries and stability constants of the proton, cobalt(II), nickel(II), copper(II) and zinc(II) complexes of 1-aminoethanephosphonic acid (α-Ala-P), 2-aminoethanephosphonic acid (β-Ala-P), 1-amino-2-phenylethanephosphonic acid (Phe-P) and 1 -amino-2-(4-hydroxyphenyl)ethanephosphonic acid (Tyr-P) have been determined pH-metrically at 25 °C and at an ionic strength of 0.2 mol dm^{− 3} (KCl).From these data and the spectral parameters of the complexes it has been established that these simple aminophosphonic acids coordinate similarly to aminocarboxylic acids, forming chelate complexes MA and MA₂. However an MAH species with only phosphonate group coordination also exist at low pH. The differences between the complex-forming properties of aminophosphonates and aminocarboxylates have been explained by the differences in basicity, charge and size of the −PO₃²⁻and −COO⁻ groups.     

Complexes of 3,4-dihydroxyphenyl derivatives. VII. Mixed ligand complexes of L-dopa and related compounds

The stability constants of the mixed ligand complexes of manganese(II), cobalt(II), nickel(II), copper(II) and zinc(II) ions with L-dopa, dopamine, L-adrenaline and L-noradrenaline as ligand A and L-alanine, L-histidine, glycylglycine and ATP as ligand B were determined pH-metrically at 25 °C and an ionic strength of 0.2 mol/dm³ (KCl). From visible spectral studies, conclusions were drawn on the bonding properties of the ambidentate ligands in the complexes.It was found that, even in its mixed ligand complexes, the ambidentate nature of dopa permits its coordination via the amino acid side-chain (N, O) at lower pH, and via the ortho phenolic hydroxy groups (O, O) at higher pH. At the same time, the complex-forming tendency of the ethanolamine side-chain in the catecholamines plays a minor role in the mixed ligand complexes.The stabilities of the mixed ligand complexes were interpreted by consideration of the differences in the stepwise stability constants of the parent complexes, the back-coordination, the charge neutralization, and the electrostatic and hydrophobic interactions between the ligand pairs.     

Formation of copper-bleomycin complexes: evidence of a three-step process

The formation of Cu(II)-bleomycin complexes as a function of pH has been studied using circular dichroism, absorption, electron paramagnetic resonance spectroscopy, and potentiometric titration. Our data support the following points: the formation of Cu(II)-bleomycin complexes occurs in a three-step process: a first complex (I) is formed at pH 1.2, which most probably involves the pyrimidine nitrogen, the secondary amine nitrogen, and two water molecules as the four in-plane ligands of copper. A second complex (II) is formed at pH 2.5, through the further coordination of the peptide nitrogen of histidine residue, and histidine imidazole nitrogen giving rise to the release of two protons. The fixation, in apical position, of the alpha-amino nitrogen of beta-aminoalanine occurs in a last step through the release of one additional proton. A value of 2.7 has been obtained for the pK of formation of this third complex, which is the species present at physiological pH. In the Cu(II)-depbleomycin system only one complex (II') has been detected.     

Coordination abilities of substituted β-aminophosphonates towards Cu and Zn ions

The formation constants of equimolar and bis-chelate copper(II) and zinc(II) complexes with three aliphatic and four aromatic-substituted β-aminophosphonates have been determined in water solution by potentiometric studies. Spectroscopic parameters clearly indicate involvement of {NH₃, PO₃ ²⁻} in both metal ions coordination. The comparison of the stability constants reveals slightly higher coordination power of the aliphatic-substituted β-aminophosphonic acids, which may be due to the higher basicity of their amino groups. All studied ligands are more effective in Cu²⁺ and Zn²⁺ coordination than phosphonic analogue of simple β-amino acid.     

Studies on the protonation constants and solvation of alpha-amino acids in dioxan-water mixtures

To gain more information about the effect of solvent on alpha-amino acids, the stoichiometric protonation constants of 10 alpha-amino acids (glycine, DL-alanine, DL-valine, L-leucine, L-isoleucine, DL-phenylalanine, L-serine, L-threonine, L-asparagine, and L-glutamine) in different dioxan-water mixtures have been determined potentiometrically using a combined pH electrode system calibrated in concentration units of hydrogen ion at 25 degrees C with an ionic strength of 0.10 M. For all amino acids studied, it was observed that the log K(1) values relating to the protonation equilibria of the anionic form are almost unaltered by the change in solvent composition. However, the log K(2) values corresponding to the formation equilibria of cationic form increase with the increase in dioxan content. The variation of these constants is discussed on the basis of specific solute-solvent interactions and structural changes of amino acids from water to dioxan-water media. The zwitterionic to neutral form ratio of these acids in dioxan-water mixtures is also discussed.     

The ionization behavior of bile acids in different aqueous environments

The ionization behavior of cholic acid, deoxycholic acid, and chenodeoxycholic acid in a variety of physiologically important molecular environments was studied using 13C NMR spectroscopy. The apparent pKa of the carboxyl group was determined from titration curves obtained from the dependence of the carboxyl carbon chemical shift on pH. Using 90% 13C isotopic substitution of the carboxyl carbon, a complete titration curve was obtained for cholate at a concentration below its critical micelle concentration and solubility limit in water. Incorporation of 12 mole % bile acid into mixed micelles with its taurine conjugate prevented precipitation of the unconjugated bile acid, and titration curves for cholic, deoxycholic, and chenodeoxycholic acids in the mixed micelles were obtained. The apparent pKa was also determined for 13C-enriched bile acids complexed with bovine serum albumin and in egg phosphatidylcholine vesicles. For monomers, micelles, and BSA complexes of all three bile acids and for deoxycholic and chenodeoxycholic acid in vesicles, one magnetic environment was observed. In contrast, two environments, both titratable, were detected for cholic acid in phosphatidylcholine vesicles. The apparent pKa's of the bile acids in the different environments ranged from 4.2 to 7.3. At pH 7.4, as monomers or bound to albumin, the bile acids were fully ionized, but when associated with phosphatidylcholine vesicles they were only partially ionized. In addition, aspects of the molecular motion and relative hydrophobicity of the bile acid carboxyl group in the environments studied were discerned from chemical shift, line-width, and lineshape data.     

Thermodynamic and structural characterization of the macrochelates formed in the reactions of copper(II) and zinc(II) ions with peptides of histidine

Copper(II) complexes of the peptides Ac-HisSarHis-NH₂, Ac-HisSarHisSarHis-NH₂ and Ac-HisSarHisSarHisSarHis-NH₂ have been studied by potentiometric, UV-Vis, CD and EPR spectroscopic methods. Stability constants for the corresponding zinc(II) complexes have also been reported. The formation of M(II)-2N_im, M(II)-3N_im and M(II)-4N_im bonded macrochelates was suggested in the pH range 5-7. The macrochelates were, however, not stable enough to prevent metal ion hydrolysis in slightly alkaline solutions. In the case of copper(II) complexes, the metal ion promoted deprotonation and coordination of the amide groups of histidyl residues were also suggested. The stability constants of macrochelate complexes were compared to the literature data reported for the macrochelates of the other peptides of histidine. It was found that the thermodynamic stability of macrochelate species is largely influenced by the number and location of histidyl residues in the peptide backbone. The highest stability was obtained for the HXHYH-type sequences, while the distant arrangement of histidyl residues resulted in a significant reduction of the stability constants.     

Hg(II)-coordination by sugar-acids: role of the hydroxy groups

A solution study on the ability of some derivatised sugars [glucuronic acid (GluA), galacturonic acid (GalA) and glucosaminic acid (GlNA)] to complex the Hg(II) ion is reported. The stability constants of the complex species were determined by potentiometric measurements while (1)H NMR experiments allow to define the coordination sites of sugar molecules. GluA coordinates the metal ion through the carboxylic oxygen and the O-4 hydroxyl group and is found to form more stable complexes with respect to GalA in which metal ligation is from the carboxylic oxygen and the O-5 ring oxygen. GlNA forms stable complexes chelating Hg(II) ion through carboxylic oxygen and the alpha-amino group. The ternary 2,2'-bipyridine containing systems were also investigated by means of potentiometric studies. The ML(2) complexes were also isolated in the solid state and characterised by IR spectroscopy.     

Ternary copper(II) complexes of levofloxacin and phenanthroline derivatives: in-vitro antibacterial, DNA interactions, and SOD-like activity

A series of ternary copper(II) complexes have been derived using levofloxacin and five phenanthroline derivatives. Complexes were characterized using infrared spectroscopy, Thermogravimetric (TG)-analysis, fast atom bombardment mass spectroscopy and reflectance spectra. Synthesized complexes exhibit the only d-d band at ～ 666?nm points toward a distorted square pyramidal geometry at metal centre with one unpaired electron responsible for paramagnetic behaviour of whole moiety. Binding behaviour of the complexes toward Herring Sperm DNA were determined using ultraviolet-Vis (UV-Vis) absorption titration and viscometric titration experiment, where as the cleavage efficacy of the complexes toward pUC19 DNA was determined by electrophoresis in presence of ethidium bromide. Complexes exhibit superoxide dismutase-like activity with their IC(50) values ranging from 0.7917 to 1.7432 μM.     

The computed distribution of copper(II) and zinc(II) ions among seventeen amino acids present in human blood plasma

The equilibrium distribution of copper(II) and zinc(II) ions among a mixture of 17 amino acids has been computed from stability-constant and blood-plasma-composition data. At pH7.4, 98% of the copper(II) in the simulated plasma solution is co-ordinated to histidine and cystine, predominantly as the mixed-ligand complexes [Cu.His.Cystine](-) and [Cu.H.His.Cystine]. Approximately half of the zinc(II) is co-ordinated to cysteine and histidine, but appreciable complex-formation occurs with most of the other amino acids. Stability constants are given for copper(II) and zinc(II) amino acid complexes, including some mixed-ligand species, at 37 degrees C and I=0.15m.