Highly efficient phosphodiester hydrolysis promoted by a dinuclear copper(II) complex

The interaction of Cu(II) with the ligand tdci (1,3,5-trideoxy-1,3,5-tris(dimethylamino)-cis-inositol) was studied both in the solid state and in solution. The complexes that were formed were also tested for phosphoesterase activity. The pentanuclear complex [Cu(5)(tdciH(-2))(tdci)(2)(OH)(2)(NO(3))(2)](NO(3))(4).6H(2)O consists of two dinuclear units and one trinuclear unit, having two shared copper(II) ions. The metal centers within the pentanuclear structure have three distinct coordination environments. All five copper(II) ions are linked by hydroxo/alkoxo bridges forming a Cu(5)O(6) cage. The Cu-Cu separations of the bridged centers are between 2.916 and 3.782 A, while those of the nonbridged metal ions are 5.455-5.712 A. The solution equilibria in the Cu(II)-tdci system proved to be extremely complicated. Depending on the pH and metal-to-ligand ratio, several differently deprotonated mono-, di-, and trinuclear complexes are formed. Their presence in solution was supported by mass, CW, and pulse EPR spectroscopic study, too. In these complexes, the metal ions are presumed to occupy tridentate [O(ax),N(eq),O(ax)] coordination sites and the O-donors of tdci may serve as bridging units between two metal ions. Additionally, deprotonation of the metal-bound water molecules may occur. The dinuclear Cu(2)LH(-3) species, formed around pH 8.5, provides outstanding rate acceleration for the hydrolysis of the activated phosphodiester bis(4-nitrophenyl)phosphate (BNPP). The second-order rate constant of BNPP hydrolysis promoted by the dinuclear complex (T = 298 K) is 0.95 M(-1) s(-1), which is ca. 47600-fold higher than that of the hydroxide ion catalyzed hydrolysis (k(OH)). Its activity is selective for the phosphodiester, and the hydrolysis was proved to be catalytic. The proposed bifunctional mechanism of the hydrolysis includes double Lewis acid activation and intramolecular nucleophilic catalysis.     

A copper(II) complex of 1,4,7-triazacyclononane featuring acetate pendant: an efficient DNA cleavage agent

A new water-soluble copper(II) complex, Cu(TACNA)Br?·?0.375H₂O (1) [TACNA?=?1,4,7-triazacyclononane-N-acetate], has been synthesized to serve as artificial nucleases. The X-ray crystal structure of 1 indicates that one bromide and an oxygen from acetate pendant coordinate to copper(II) in addition to the nitrogen atoms in the TACN macrocycle, resulting in a five-coordinate complex with square-pyramidal geometry. The interaction of 1 with calf thymus DNA (ct-DNA) has been investigated by UV absorption and fluorescence spectroscopies, and the mode of ct-DNA binding for 1 has been proposed. In the absence of external agents, supercoiled plasmid DNA cleavage by 1 was performed under aerobic condition; the influences on DNA cleavage of different complex concentrations and reaction times were also studied. The cleavage of plasmid DNA likely involves oxidative mechanism.     

Poly(ethylene glycol)-supported copper(II) triazacyclononane: an efficient, recoverable, and recyclable catalyst for the cleavage of a phosphodiester

The triazacyclononane macrocycle has been linked to the soluble polymer monomethoxy poly(ethylene) glycol and its copper(II) complex efficiently catalyzes the hydrolysis of a model phosphodiester. The catalyst, easily recovered from the water solution, shows no appreciable loss of activity when recycled three times.     

A simple copper(II)-L-histidine system for efficient hydrolytic cleavage of DNA

Copper(II)-L-histidine complexes effectively promote the cleavage of plasmid DNA and dideoxynucleotide dApdA at physiological pH and temperature. Studies of the mechanism of plasmid DNA cleavage by added radical scavengers, using rigorously anaerobic experiments, analyses for malondialdehyde-like products, religation assays, and HPLC analyses, indicate that DNA cleavage mediated by Cu(L-His) occurs via a hydrolytic path. The hydrolytic cleavage rate constants at 37 degrees C are estimated to be 0.76 h-1 for the decrease of form I and 0.25 h-1 for the increase of form III. The phosphoimager picture reveals that Cu(L-His) cleaves DNA with a certain sequence specificity (preferentially at 5'-GT-3'). The dinucleotide hydrolysis shows, with [Cu(L-His)] = 0.8 mM, rate enhancement factors of > 10(8). Interestingly, histidine-metal ion interactions (with Cu(II), Ni(II), Zn(II), etc.) have been used for various applications, e.g., protein purification, cross-linking, and targeting proteins to lipid bilayers. Our findings may provide the basis for developing new applications and new ways to design more effective and useful catalysts for DNA cleavage. Cu(L-His) is one of only a few well-defined metal complexes demonstrated to hydrolytically cleave dideoxynucleotides and DNA.     

A new copper(II) complex as an efficient catalyst of luminol chemiluminescence

A new copper complex (2) has exhibited highly efficient catalytic activity of luminol chemiluminescence in water in the presence of ascorbic acid and dissolved O2 under conditions that conventional catalysts such as Cu(OAc)2, hemin or cyclen-Cu(II) did not show significant activity.     

Potential-modulated DNA cleavage by (N-salicylideneglycinato)copper(II) complex.

The interaction of aqua (N-salicylideneglycinato)copper(II) (Cu(salgly)2+) complex with calf thymus DNA has been investigated by cyclic voltammetry. Potential-modulated DNA cleavage in the presence of Cu(salgly)2+ complex was performed at a gold electrode in a thin layer cell. DNA can be efficiently cleaved by electrochemically reducing Cu(salgly)2+ complex to Cu(salgly)+ complex at -0.7 V (vs. Ag/AgCl). When the solution was aerated with a small flow of O2 during electrolysis, the extent of DNA cleavage was dramatically enhanced, and hydroxyl radical scavengers inhibited DNA cleavage. These results suggested that O2 and hydroxyl radical were involved in potential-modulated DNA cleavage reaction. The percentage of DNA cleavage was enhanced as the working potential was shifted to more negative values and the electrolysis time was increased. It was also dependent on the ratio of Cu(salgly)2+ complex to DNA concentration. The cleaved DNA fragments were separated by high performance liquid chromatography (HPLC). The experimental results indicated that the method for potential-modulated DNA cleavage by Cu(salgly)2+ complex was simple and efficient.     

Intramolecular nucleophilic activation promoting efficient hydrolytic cleavage of DNA by (aqua)bis(dipyridoquinoxaline)copper(II) complex

The axial aqua bound copper(II) complex [Cu(dpq)2(H2O)](ClO4)2, having a planar NN-donor heterocyclic base dipyridoquinoxaline (dpq) as the DNA minor groove binder, shows efficient hydrolytic cleavage of supercoiled DNA in the dark and in the absence of any external reagents, as evidenced from T4 ligase experiments, with a rate of 5.58 +/- 0.4 h(-1) and a rate enhancement of 1.55 x 10(8).     

Efficient plasmid DNA cleavage by a mononuclear copper(II) complex

The Cu(II) complex of the ligand all-cis-2,4,6-triamino-1,3,5-trihydroxycyclohexane (TACI) is a very efficient catalyst of the cleavage of plasmid DNA in the absence of any added cofactor. The maximum rate of degradation of the supercoiled plasmid DNA form, obtained at pH 8.1 and 37 degrees C, in the presence of 48 microM TACI.Cu(II), is 2.3 x 10(-3) s(-1), corresponding to a half-life time of only 5 min for the cleavage of form I (supercoiled) to form II (relaxed circular). The dependence of the rate of plasmid DNA cleavage from the TACI.Cu(II) complex concentration follows an unusual and very narrow bell-like profile, which suggests an high DNA affinity of the complexes but also a great tendency to form unreactive dimers. The reactivity of the TACI.Cu(II) complexes is not affected by the presence of several scavengers for reactive oxygen species or when measured under anaerobic conditions. Moreover, no degradation of the radical reporter Rhodamine B is observed in the presence of such complexes. These results are consistent with the operation of a prevailing hydrolytic pathway under the normal conditions used, although the failure to obtain enzymatic religation of the linearized DNA does not allow one to rule out the occurrence of a nonhydrolytic oxygen-independent cleavage. A concurrent oxidative mechanism becomes competitive upon addition of reductants or in the presence of high levels of molecular oxygen: under such conditions, in fact, a remarkable increase in the rate of DNA cleavage is observed.     

Metal-assisted red light-induced efficient DNA cleavage by dipyridoquinoxaline-copper(II) complex

Complete cleavage of double stranded pUC19 DNA by the complex [Cu(dpq)2(H2O)](ClO4)2 (dpq, dipyridoquinoxaline) has been observed on irradiation at 694 nm from a pulsed ruby laser, assisted by the metal d-band transition as well as the quinoxaline triplet states in the absence of any external additives.     

A highly reactive mononuclear Zn(II) complex for phosphodiester cleavage

The activity of a Zn(II) complex of a tetradentate, tripodal ligand for catalyzing phosphodiester cleavage is enhanced 750-fold by introducing three hydrogen bond donors to the ligand. Inhibition studies show that the Zn-aqua complex is the kinetically active form and that it binds the transition state with a formal dissociation constant of 3 x 108 M-1. The effect of these ligand modifications on the transition-state affinity is comparable to the rate acceleration provided by the metal ion itself. Overall, this mononuclear complex is more active than the most reactive dinuclear Zn(II) complexes reported to date.     

Polymer‐anchored copper(II) complex: an efficient reusable catalyst for the synthesis of propargylamines

A new, heterogeneous, polymer‐supported copper(II) complex was prepared and characterized using various techniques, including Fourier transform infrared spectroscopy, scanning electron microscopy, energy dispersive X‐ray spectroscopy, atomic absorption spectroscopy and thermogravimetric analysis. This heterogeneous copper catalyst is efficient for the synthesis of propargylamines via a three‐component coupling reaction of aldehydes, amines and alkynes. The effect of solvent on the coupling reactions was investigated. Further, the catalyst can be easily recovered quantitatively by simple filtration and reused for a minimum number of cycles without significant loss of its catalytic activity. Copyright © 2014 John Wiley & Sons, Ltd.     

Synthesis,DNA binding and oxidative cleavage studies of an asymmetric tridentate copper(II) complex

An asymmetric tridentate Cu^II complex, [Cu(ptp)Cl₂] (ptp = 3-(1,10-phenanthrolin-2-yl)-as-triazino[5,6-f]-phenanthrene), has been synthesized and characterized by elemental analysis, FAB-MS, i.r. and u.v.–vis. spectra. The DNA binding behavior of the complex has been examined by fluorescence quenching, cyclic voltammetric and viscosity measurements. The results suggest that [Cu(ptp)Cl₂] binds to DNA in the intercalative mode. This complex is also found to produce cleavage of pBR 322 DNA in the presence of reducing agents such as ascorbate/H₂O₂, and the hydroxyl radical (OH^•) is suggested to be the reactive species responsible for the cleavage.     

Syntheses,characterization and DNA cleavage studies of acyclic copper(II) complexes

Two novel acyclic copper(II) complexes, Cu(L₁) (1) {L₁ = N,N-1,2-ethanediylbis[N-(phenylmethyl)glycinato]} and Cu(L₂) (2), {L₂ = N,N-1,2-ethanediylbis[N-[(3-nitrophenyl)methyl]glycinato]} have been synthesized and characterized by elemental analysis and ES-MS. Thermal denaturation, fluorescence spectroscopy and cyclic voltammetry have been conducted to assess the interaction of the two complexes with calf thymus DNA. Interestingly, the two copper(II) complexes have been found to cleave circular plasmid pBR322 DNA to the nicked form (Form II) and the linear form (Form III) under aerobic conditions.     

Solvent deuterium isotope effects on phosphodiester cleavage catalyzed by an extraordinarily active Zn(II) complex

The effect of increasing pL on the extraordinary catalytic activity of a dinuclear Zn2+ complex toward cleavage of uridine 3'-4-nitrophenyl phosphate (UpPNP) in H2O and D2O was determined. This change from H2O to D2O causes an increase from 7.8 to 8.4 in the apparent pKa of a catalytic functional group, but has little effect on the activity of the active form of the catalyst toward cleavage of UpPNP, so that there is no primary kinetic SDIE on the cleavage reaction from movement of a proton at the rate-determining transition state. It is concluded that essentially all of the rate acceleration for this catalyst is due to electrostatic stabilization of the transition state by interactions between opposing cationic and anionic charges.     

A common chemical mechanism used for DNA cleavage by copper(II) activated by thiols and ascorbate is distinct from that for copper(II): hydrogen peroxide cleavage

Summary By use of a [³²P] end-labelled 163 base pair linear fragment of duplex DNA isolated from tyrTDNA, the apparent sequence selectivity previously reported for DNA cleavage by Cu^II:thiol systems was shown to be identical to the sequence selectivity in scission shown by the Cu^II:ascorbate system. On this basis, a general mechanism of DNA cleavage is proposed for Cu^II with reducing agents including thiols as well as reducing sugars, including ascorbate and glucosamine. This mechanism involves hydroxyl radical generation and leads to both base damage and strand scission, and based on results with Cu^II:thiol cleavage the former occurs before the latter is detectable. Cleavage of DNA by Cu^II:hydrogen peroxide follows a different chemical mechanism, distinguishable by its different sequence preference.     

DNA binding, prominent DNA cleavage and efficient anticancer activities of tris(diimine)iron(II) complexes

The complexes rac-[Fe(diimine)(3)](ClO(4))(2)1-4, where diimine = 2,2'-bipyridine (bpy) 1, 1,10-phenanthroline (phen) 2, 5,6-dimethyl-1,10-phenanthroline (5,6-dmp) 3 and dipyrido[3,2-d:2',3'-f]quinoxaline (dpq) 4, have been isolated, characterized and their interaction with calf thymus DNA studied by using a host of physical methods. The X-ray crystal structure of rac-[Fe(5,6-dmp)(3)](ClO(4))(2)3 has been determined and the packing diagram shows the presence of two enantiomeric forms of the complex cations in the same unit cell. The structures of 1-4 in solution have also been studied using UV-Visible, Cyclic Voltammetry and ESI-MS data and all data available suggests that they retain their solid state structures even in solution. The absorption spectral titrations of the iron(ii) complexes with CT DNA reveal that the DNA binding affinities of the complexes vary in the order, 4 (K(b): 9.0 × 10(3)) > 2 (6.8 × 10(3)) > 3 (4. 8 × 10(3)) > 1 (2.9 × 10(3) M(-1)). The DNA interaction of dpq complex (4) involves partial insertion of the extended phen ring in between the DNA base pairs, which is deeper than that of phen (2). The 5,6-dmp (3) complex is involved in groove binding in the major groove of DNA. The lower DNA binding affinity of 1 is due to electrostatic interaction of the cationic complexes with exterior phosphates of DNA. The EthBr displacement assay and DNA viscosity study support these DNA binding modes and the above trend in DNA binding affinities. The complexes of 1 and 2 show induced CD (ICD) upon interaction with CT DNA while 3 and 4 bound to DNA exhibit inversion in the positive band with the helicity band showing very small changes, which implies that 3 and 4 bind enantiopreferentially to DNA. The DNA cleavage abilities of 1-4 have been observed at 10 μM concentration of complexes in the presence of 100 μM H(2)O(2) and the DNA cleavage efficiency (> 90%) follows the order 3 > 1 > 2 > 4. The anticancer activity of 1-4 against human breast cancer cell line (MCF-7) has also been studied. The IC(50) values of the complexes at different incubation time intervals of 24 and 48 h follow the order, 3 (0.8, 0.6) < 4 (20.0, 17.0) < 2 (28.0, 22.0) < 1 (32.0, 29.0 μM). Interestingly, 3 exhibits anticancer activity more potent than 1, 2 and 4 and cisplatin for both 24 and 48 h. It induces cell death both through apoptosis and necrosis mechanisms, as revealed by morphological assessment data obtained by using AO/EB and Hoechst 33258 fluorescence staining methods.     

Red-light photosensitized cleavage of DNA by (l-lysine)(phenanthroline base)copper(II) complexes

Ternary copper(II) complexes [Cu(l-lys)B(ClO4)](ClO4)(1-4), where B is a heterocyclic base, viz. 2,2'-bipyridine (bpy, 1), 1,10-phenanthroline (phen, 2), dipyrido[3,2-d:2',3'-f]quinoxaline (dpq, 3) and dipyrido[3,2-a:2',3'-c]phenazene (dppz, 4), are prepared and their DNA binding and photo-induced DNA cleavage activity studied (l-lys =l-lysine). Complex 2, structurally characterized by X-ray crystallography, shows a square-pyramidal (4 + 1) coordination geometry in which the N,O-donor l-lysine and N,N-donor heterocyclic base bind at the basal plane and the perchlorate ligand is bonded at the elongated axial site. The crystal structure shows the presence of a pendant cationic amine moiety -(CH2)4NH3+ of l-lysine. The one-electron paramagnetic complexes display a d-d band in the range of 598-762 nm in DMF and exhibit cyclic voltammetric response due to Cu(II)/Cu(I) couple in the range of 0.07 to -0.20 V vs. SCE in DMF-Tris-HCl buffer. The complexes having phenanthroline bases display good binding propensity to the calf thymus DNA giving an order: 4 (dppz) > 3 (dpq) > 2 (phen)> 1 (bpy). Control cleavage experiments using pUC19 supercoiled DNA and distamycin suggest major groove binding for the dppz and minor groove binding for the other complexes. Complexes 2-4 show efficient DNA cleavage activity on UV (365 nm) or visible light (694 nm ruby laser) irradiation via a mechanistic pathway involving formation of singlet oxygen as the reactive species. The amino acid l-lysine bound to the metal shows photosensitizing effect at red light, while the heterocyclic bases are primarily DNA groove binders. The dpq and dppz ligands display red light-induced photosensitizing effects in copper-bound form.     

Studies of copper (Ⅱ) complexes as catalysts for the hydrolysis of DNA

Hydrolysis of DNA is an important enzymatic reaction , but it is exceedingly difficult to mimic in the laboratory because of the stability of hydrolysis of DNA. In this paper, the cleavage activity of complexes formed between Cu(Ⅱ) and four different amino acid or amino acid methyl ester on DNA is studied by gel elec-trophoresis. It is found that DNA could be cleaved by Cu(Ⅱ)-L-His and Cu(Ⅱ)-L-His methyl ester complexes and the efficiency of cleavage is largely dependent on the metal ion-to-ligand ratio. Further experiments show that the cleavage of DNA mediated by Cu(Ⅱ)-L-His complexes occurs via a hydrolytic mechanism and the active chemical species that affects DNA cleavage is proposed to be MI2H and ML2H22 .     

八羟基喹啉铜(Ⅱ)配合物的DNA结合和切割活性及与牛血清白蛋白相互作用（英文）

利用紫外吸收光谱、荧光光谱、圆二色光谱(CD)和琼脂糖凝胶电泳等手段研究了八羟基喹啉铜(Ⅱ)配合物Cu[8-OHQ]2与DNA和蛋白质的相互作用.实验结果表明,在生理条件下,Cu[8-OHQ]2能通过插入方式较强的与CT-DNA结合,诱导DNA构象的改变.其本征结合常数Kb为1.15(±0.01)×105 L/mol,表观结合常数Kapp为4.21×106 L/mol.再者,琼脂糖凝胶电泳实验表明,在生理条件和抗坏血酸(Vc)存在情况下,Cu[8-OHQ]2能有效地将超螺旋pBR322质粒DNA切割成缺刻和线性,甚至降解为小的片断.机理研究表明扩散的·OH,H2O2和1 O2都不是在切割过程中起作用的活性氧物种(ROS);copper-oxo中间体可能是此切割过程中主要的活性氧物种.另外,Cu[8-OHQ]2也能以适中的结合力与牛血清白蛋白(BSA)结合而猝灭BSA内源荧光,猝灭机理为静态猝灭.所有这些结果表明Cu[8-OHQ]2具有作为潜在化疗试剂的生物活性.     

Zinc(II) oxide: an efficient catalyst for selective transesterification of β-ketoesters

A facile and selective transesterification of β-ketoesters using zinc(II) oxide as catalyst is described. The emphasis has been placed on the reaction of methyl 3-(3,4-dimethoxyphenyl)-3-oxopropanoate with a series of alcohols of different structures, leading in all cases good to excellent yields.