Study of the Intramolecular Reaction of an Activated Phosphate Ester in the Micelle System Containing Macrocyclic Complex

The intramolecular nucleophilic substitution of an activated phosphate diester, bis(p-nitrophenyl) phosphate (BNPP) as the nucleic acids substitute, was investigated. A macro-cyclic ligand and the corresponding Cu (II) and Ni (II) complexes were synthesized and characterized. The metallomicelles made up of macrocyclic divalent metal complex and micelle, as mimic hydrolytic metalloenzyme, was used in BNPP catalytic hydrolysis. The metallomicelles displayed higher catalytic activity although they do not attain the catalytic efficiency of enzymes. The analysis of specific absorption spectra showed that the course of the BNPP catalytic reaction was different from that of the BNPP spontaneous hydrolysis, and was an intramolecular nucleophilic substitution reaction. Based on the analytic result of the specific absorption spectrum, an intramolecular nucleophilic substitution mechanism of BNPP catalytic hydrolysis was proposed and a correlative kinetic mathematical model was established, and the corresponding thermodynamic and kinetic constant was calculated. The result of this study proved validity of the mechanism and mathematical model proposed in the article.     

Metallomicellar catalysis: accelerated hydrolysis of BNPP by copper(II), zinc(II), and nickel(II) complexes of long alkanol-imidazole in CTAB micellar solution

Two alkanol-imidazole ligands have been synthesized. Metal (Cu(II), Zn(II), Ni(II)) complexes of these ligands have been investigated as catalysts for the hydrolysis of bis(p-nitrophenyl) phosphate (BNPP) in the buffered CTAB co-micellar solution at 30 degrees C and various pH values, respectively. The ternary complex kinetic model for metallomicellar catalysis was employed to analyze the results, to obtain the kinetic and thermodynamic parameters. The effect of the structure of the ligands and the microenvironment of the reaction on the hydrolytic reaction of BNPP has been discussed in detail. The results indicate that the hydrophobic interaction between the substrate and the metallomicellar catalyst and the micellar microenvironment are important factors.     

Hydrolysis of Bis(4‐Nitrophenyl) Phosphate Catalyzed by Metallomicelle Made Up of the Crowned Schiff Base Complex as Synthetic Hydrolase

A crowned Schiff base ligand and its cobalt(II) and manganese(III) complexes were synthesized and characterized, and the metallomicelles made from the complexes and micelles (Brij35, LSS, CTAB) were investigated to catalyze the hydrolysis of bis(4‐nitrophenyl) phosphate (BNPP). A kinetic mathematical model for simulating enzyme catalyzing reaction was proposed and employed to analyze the mechanism of BNPP catalytic hydrolysis. Michanelis constant and the apparent active energy for the catalytic reaction were calculated. The kinetic studies showed that the metallomicelle made from the micelle and crowned Schiff base transitional metal complexes is an effective mimetic hydrolytic enzyme for BNPP catalytic hydrolysis.     

Hydrolysis of phosphodiester catalyzed by analogous dinuclear Cu(II) complex in CTAB micellar solution

The catalytic hydrolysis of bis(p-nitrophenyl) phosphate (BNPP) by metallomicelle composed of analogous Cu(II) complex in CTAB micellar solution was investigated at 30 °C and different pH. The results indicate that the analogous complex with 1:2 ratio of ligand to metal ion in CATB micellar solution is the active species for catalyzing the hydrolysis of BNPP. The ternary complex kinetic model for metallomicellar catalysis was employed to obtain the relative kinetic and thermodynamic parameters for taking apart the mechanism of the catalytic hydrolysis of BNPP.     

Metallomicellar Catalytic Hydrolysis of NPP by CuIINiII Heterodinuclear Complexes Containing Diamine Groups in Brij35 Micellar Solution

The hydrolysis of p‐nitrophenyl phosphate (NPP) catalyzed by metallomicelles formed from four asymmetry Cu^IINi^II heterodinuclear oxamido‐bridged complexes, whose ligands contain different diamine groups, and nonionic surfactant Brij35 has been investigated quantitatively at the pH range of 6.0–10.5 and 25°C, respectively. The results indicated that the rates of catalytic hydrolysis of NPP by four complexes in Brij35 micellar solution were all largely enhanced under the experimental conditions. The order of effectiveness of the four complexes in Brij35 micellar solutions toward hydrolysis of NPP was found to be: complex I>complex II>complex III>complex IV. The different catalytic function of four complexes may be attributed to the different activation abilities of coordinated water molecules induced by different polarities of ligands, which has been discussed in detail. Meanwhile, kinetic studies show that the active species in the catalytic hydrolysis of NPP may be the aquo‐hydroxy form and that the mechanism of the reaction involves intramolecular nucleophilic attack of the nickel‐bound hydroxide ion on phosphoryl of NPP. The studies suggested that the metallomicelle containing Cu^IINi^II heterodinuclear oxamido‐bridged complexes with two coordinated H₂O molecules may be a potential catalyst for the hydrolysis of NPP.     

Studies on the reaction kinetics and the mechanism of hydrolysis of bis(4-nitrophenyl) phosphate (BNPP) catalyzed by oxamido-bridged dinuclear copper(II) complexes in micellar solution

Oxamido-bridged dinuclear copper(II) complexes, used as symmetric two-center catalysts for the cleavage of BNPP, were synthesized and characterized. The reaction kinetics and the mechanism of hydrolysis of BNPP catalyzed by metallomicelles, made from complex (A) or (B) and a surfactant (LSS or CTAB), were investigated. A kinetic mathematical model for BNPP cleavage was also proposed. The results showed that the reaction rate for the catalytic hydrolysis of BNPP, compared with spontaneous hydrolysis of BNPP, increased by a factor of ca. 1 × 10⁶ due to the synergistic effect of two copper ions in the complex and the local concentration effect of the micelle. The study indicates that the metallomicelle-containing oxamido-bridged dinuclear copper(II) complex may be a potential catalyst for the hydrolysis of BNPP.     

Metallomicellar Catalysis Hydrolysis of p-Nitrophenyl Picolinate Catalyzed by Copper(II), Nickel(II), and Zinc(II) Complexes of Long Alkyl Pyridine Ligands in Micellar Solution

The ternary complex kinetic model for metallomicellar catalysis has been proposed in this paper. The catalytic effects of bivalent metal ion (Cu(2+), Zn(2+), and Ni(2+)) complexes of long alkyl pyridine ligands upon the hydrolysis of p-nitrophenyl picolinate (PNPP) have been studied kinetically in aqueous buffer of pH ranging from 5.0 to 8.5 at 30 degrees C. The effect of pH on the reactivity is discussed. The results indicate that the metallomicelles formed by pyridine ligands promote the hydrolysis of PNPP, and the order is Cu(II) system>Ni(II) system>Zn(II) system. A stereochemical modification of the complex in the CTAB micelle is suggested as a likely explanation for the observed phenomena. Copyright 2000 Academic Press.     

Metallomicellar Catalysis: Hydrolysis of Phosphodiester with Cu(II) and Zn(II) Complexes in Micellar Solution

Abstract

The effects of two metal complexes of 2,2′‐dipyridylamine (bpya) ligand, [(bpya)Cu]Cl₂ and [(bpya)Zn]Cl₂, in promoting the hydrolysis of bis(4‐nitrophenyl) phosphate (BNPP) have been kinetically investigated in Brij35 micellar solution and at 298 K, pH ranging from 6.41 to 8.6. In neutral micellar solution at 298 K, pH 7.02, the rate constants for the catalytic hydrolysis of BNPP by [(bpya)Cu]Cl₂ and [(bpya)Zn]Cl₂ are 1.2 × 10⁶ times and 1.5 × 10⁵ times higher than those for the spontaneous hydrolysis, respectively. Kinetic studies show that the active species in the catalytic hydrolysis of BNPP is the aquo‐hydroxy form, and the relative kinetic and thermodynamic parameters indicate that the mechanism of the reaction involves intramolecular nucleophilic attack on the metal center‐bound diester.     

Studies on BNPP Cleavage by Schiff Base Complexes Containing Benzoaza‐15‐Crown‐5 in DHAB Micellar Solution

Two novel benzoaza‐crown Schiff base cobalt (II) and manganese (III) complexes were synthesized and characterized. The hydrolysis of bis(4‐nitrophenyl) phosphate (BNPP) catalyzed by the two complexes was studied in buffer solution containing dihexadecyldimethylammonium bromide (DHAB) at 25°±0.1°C and different pH values. The kinetic mathematical model of BNPP hydrolysis was proposed, and the effects of different reaction conditions on BNPP hydrolysis were discussed. The results indicate that the two complexes (MnLCl and CoL) can efficiently accelerate the catalytic cleavage of BNPP in DHAB micellar solution. The pseudo‐first‐order rate constants (k _obsd) of BNPP hydrolysis catalyzed by the metallomicelles of MnLCl/DHAB and CoL/DHAB are 2.32×10⁷ times and 1.45×10⁷ times higher than that of the BNPP spontaneous hydrolysis, respectively. Possible reasons for the huge rate accelerations include the lower critical micelle concentration (cmc) of DHAB and formation of metallomicelles made of complexes and DHAB. Furthermore, the BNPP cleavage catalyzed only by the two complexes was investigated in buffer solution. It was found that the hydrolytic rates of BNPP catalyzed only by the two complexes were about 1% of those catalyzed by MnLCl/DHAB and CoL/DHAB systems at 25°C, pH=7.00, and [BNPP]=2.0×10^?4 mol · dm^?3.     

Comparative Reactivity of Phosphate Ester Hydrolysis Catalyzed by Mononuclear and Hetero-Dinuclear Complexes Containing the Lanthanum Ion (III)

The hydrolysis of bis(p-nitrophenyl phosphate)(BNPP) and p-nitrophenyl phosphate (NPP) were strongly promoted by the mononuclear La^III[(N,N-bis(3-carboxyl)-2-hydroxysalicylidene)ethylenediamine] and the hetero-dinuclear Cu^IILa^III[(N,N-bis(3-carboxyl)-2-hydroxysalicylidene)ethylenediamine] complexes at 35 °C in 50% aqueous DMSO. The results indicated that the hetero-dinuclear Cu^IILa^III complex exhibited a relatively higher catalytic function on NPP hydrolysis, while the hydrolysis of BNPP was slightly more efficient in the presence of the mononuclear La^III complex than that in the presence of the Cu^IILa^III complex, and this case was discussed by the different spacer requirements for NPP and BNPP hydrolysis. In addition, widely differing pH-rate profiles were obtained: saturated curves for NPP hydrolysis and bell-shaped curves for BNPP hydrolysis, and thus different kinetic treatments were employed to obtain the relative parameters.     

Hydrolysis of DNA model substrates catalyzed by metal-substituted Wells-Dawson polyoxometalates

In this study we report the first example of phosphoester bond hydrolysis in 4-nitrophenyl phosphate (NPP) and bis-4-nitrophenyl phosphate (BNPP), two commonly used DNA model substrates, promoted by metal-substituted polyoxometalates (POMs). Different transition metal and lanthanide ions were incorporated into the Wells-Dawson polyoxometalate framework and subsequently screened for their hydrolytic activity towards the cleavage of the phosphoester bonds in NPP and BNPP. From these complexes, the Zr(iv)-substituted POM showed the highest reactivity. At pD 7.2 and 50 °C a NPP hydrolysis rate constant of 7.71 × 10(-4) min(-1) (t(1/2) = 15 h) was calculated, representing a rate enhancement of nearly two orders of magnitude in comparison with the spontaneous hydrolysis of NPP. The catalytic (k(c) = 1.73 × 10(-3) min(-1)) and formation constant (K(f) = 520.02 M(-1)) for the NPP-Zr(iv)-POM complex were determined from kinetic experiments. The reaction proceeded faster in acidic conditions and (31)P NMR experiments showed that faster hydrolysis is proportional to the presence of the 1?:?1 monosubstituted Zr(iv)-POM at acidic pD values. The strong interaction of the 1?:?1 monosubstituted Zr(iv)-POM with the P-O bond of NPP was evidenced by the large chemical shift and the line broadening of the (31)P nucleus in NPP observed upon addition of the metal complex. Significantly, a ten-fold excess of NPP was fully hydrolyzed in the presence of the Zr(iv)-POM, proving the principles of catalysis. The NMR spectra did not show sign of any paramagnetic species, excluding an oxidative cleavage mechanism and suggesting purely hydrolytic cleavage.     

Binuclear copper(II) complexes of xylyl-bridged bis(1,4,7-triazacyclononane) ligands

Copper(II) complexes of three bis(tacn) ligands, [Cu(2)(T(2)-o-X)Cl(4)] (1), [Cu(2)(T(2)-m-X)(H(2)O)(4)](ClO(4))(4).H(2)O.NaClO(4) (2), and [Cu(2)(T(2)-p-X)Cl(4)] (3), were prepared by reacting a Cu(II) salt and L.6HCl (2:1 ratio) in neutral aqueous solution [T(2)-o-X = 1,2-bis(1,4,7-triazacyclonon-1-ylmethyl)benzene; T(2)-m-X = 1,3-bis(1,4,7-triazacyclonon-1-ylmethyl)benzene; T(2)-p-X = 1,4-bis(1,4,7-triazacyclonon-1-ylmethyl)benzene]. Crystals of [Cu(2)(T(2)-m-X)(NPP)(mu-OH)](ClO(4)).H(2)O (4) formed at pH = 7.4 in a solution containing 2 and disodium 4-nitrophenyl phosphate (Na(2)NPP). The binuclear complexes [Cu(2)(T(2)-o-XAc(2))(H(2)O)(2)](ClO(4))(2).4H(2)O (5) and [Cu(2)(T(2)-m-XAc(2))(H(2)O)(2)](ClO(4))(2).4H(2)O (6) were obtained on addition of Cu(ClO(4))(2).6H(2)O to aqueous solutions of the bis(tetradentate) ligands T(2)-o-XAc(2) (1,2-bis((4-(carboxymethyl)-1,4,7-triazacyclonon-1-yl)methyl)benzene and T(2)-m-XAc(2) (1,3-bis((4-(carboxymethyl)-1,4,7-triazacyclonon-1-yl)methyl)benzene), respectively. In the binuclear complex, 3, three N donors from one macrocycle and two chlorides occupy the distorted square pyramidal Cu(II) coordination sphere. The complex features a long Cu...Cu separation (11.81 A) and intermolecular interactions that give rise to weak intermolecular antiferromagnetic coupling between Cu(II) centers. Complex 4 contains binuclear cations with a single hydroxo and p-nitrophenyl phosphate bridging two Cu(II) centers (Cu...Cu = 3.565(2) A). Magnetic susceptibility studies indicated the presence of strong antiferromagnetic interactions between the metal centers (J = -275 cm(-1)). Measurements of the rate of BNPP (bis(p-nitrophenyl) phosphate) hydrolysis by a number of these metal complexes revealed the greatest rate of cleavage for [Cu(2)(T(2)-o-X)(OH(2))(4)](4+) (k = 5 x 10(-6) s(-1) at pH = 7.4 and T = 50 degrees C). Notably, the mononuclear [Cu(Me(3)tacn)(OH(2))(2)](2+) complex induces a much faster rate of cleavage (k = 6 x 10(-5) s(-1) under the same conditions).     

Catalytic hydrolysis of phosphate diester （BNPP） and plasmid DNA by mononuclear macrocyclic polyamine metal complexes

The activities of the catalytic hydrolysis of phosphate diester(BNPP)[bis(p-nitrophenyl)phosphate diester]and plasmid DNA (pUC18)by mononuclear macrocyclic polyamine metal complexes have been investigated in this paper.The results showed that the highest activity in hydrolysis of BNPP was obtained with 1e-Zn(Ⅱ)complex(composed of lipophilic group)as catalyst.The hydrolysis rate enhancement is up to 3.64×10~4 fold.These metal complexes could effectively promote the cleavage of plasmid DNA(pUC18)at physiol...     

Zn(II) coordination to polyamine macrocycles containing dipyridine units. New insights into the activity of dinuclear Zn(II) complexes in phosphate ester hydrolysis

Zn(II) binding by the dipyridine-containing macrocycles L1-L3 has been analyzed by means of potentiometric measurements in aqueous solutions. These ligands contain one (L1, L2) or two (L3) 2,2'-dipyridine units as an integral part of a polyamine macrocyclic framework having different dimensions and numbers of nitrogen donors. Depending on the number of donors, L1-L3 can form stable mono- and/or dinuclear Zn(II) complexes in a wide pH range. Facile deprotonation of Zn(II)-coordinated water molecules gives mono- and dihydroxo-complexes from neutral to alkaline pH values. The ability of these complexes as nucleophilic agents in hydrolytic processes has been tested by using bis(p-nitrophenyl) phosphate (BNPP) as a substrate. In the dinuclear complexes the two metals play a cooperative role in BNPP cleavage. In the case of the L2 dinuclear complex [Zn(2)L2(OH)(2)](2+), the two metals act cooperatively through a hydrolytic process involving a bridging interaction of the substrate with the two Zn(II) ions and a simultaneous nucleophilic attack of a Zn-OH function at phosphorus; in the case of the dizinc complex with the largest macrocycle L3, only the monohydroxo complex [Zn(2)L3(OH)](3+) promotes BNPP hydrolysis. BNPP interacts with a single metal, while the hydroxide anion may operate a nucleophilic attack. Both complexes display high rate enhancements in BNPP cleavage with respect to previously reported dizinc complexes, due to hydrophobic and pi-stacking interactions between the nitrophenyl groups of BNPP and the dipyridine units of the complexes.     

Studies on the reaction kinetics and the mechanism of hydrolysis of bis(4-nitrophenyl)phosphate (BNPP) by hydroxamic acid complexes containing benzo-15-crown-5

A hydroxamic acid (HL) containing benzo-15-crown-5 and their copper(II), zinc(II), cobalt(II) and manganese(II) complexes have been synthesized and studied as catalysts for the cleavage of bis(4-nitrophenyl)phosphate (BNPP). The catalytic properties of these complexes and the kinetics and mechanism of BNPP hydrolysis have been investigated. The kinetic mathematical model of BNPP cleavage catalyzed by these complexes has been proposed. The effects of reaction temperature and metal ion in the complexes on the rate of BNPP catalytic hydrolysis have been discussed. The results show that the hydroxamic acid complexes containing benzo-15-crown-5 exhibit high activity in the BNPP catalyzed hydrolysis; the pseudo-first-order-rate constants of BNPP hydrolysis catalyzed by the complexes increase along with the increases of pH of the buffer solution from 7.50 to 9.50; the activity of different metal ions decreases in the order: Co²⁺ > Cu²⁺ > Zn²⁺ > Mn²⁺; the pseudo-first-order-rate constants of BNPP hydrolysis catalyzed by the complexes is 2.24 × 10⁵ ~ 3.24 × 10⁵ times as large as that of spontaneous hydrolysis of BNPP.     

Hydrolysis of BNPP Catalyzed by Metallomicelles Made of Pr(III) Complexes

A macrocyclic ligand was synthesized and characterized. The kinetics of hydrolysis of bis(p-nitrophenyl)phosphate (BNPP) in the catalytic system containing macrocyclic ligand and praseodymium(III) was investigated. The analysis of specific absorption spectrums of the hydrolytic reaction systems indicated that key intermediates made up of BNPP and praseodymium(III) complexes are formed in the reaction process of BNPP catalytic hydrolysis. In this, the mechanism of BNPP catalytic hydrolysis proposed is based on the analytic result of specific absorption spectrum, and the corresponding kinetic constants are calculated. The results showed that the praseodymium(III) complexes as hydrolase mimics exhibit good catalytic activity and similar catalytic character to natural enzyme.     

Hydrolysis of BNPP Catalyzed by the Crowned Schiff Base Co(II) Complexes in Micellar Solution

Two symmetrical double aza‐crowned Schiff base cobalt(II) complexes were synthesized and characterized, and the metallomicelle made up of the cobalt(II) complexes and surfactant, as mimic hydrolytic metalloenzyme, was used in catalytic hydrolysis of bis(4‐nitrophenyl) phosphate (BNPP). The analysis of specific absorption spectrums of the hydrolytic reaction systems indicated that key intermediates made up of BNPP and Co(II) complexes are formed in reaction processes of the BNPP catalytic hydrolysis. In this article, the mechanism of BNPP catalytic hydrolysis has been proposed based on the analytic result of specific absorption spectrum. A kinetic mathematical model, for the calculation of the kinetic parameter of BNPP catalytic hydrolysis has been established based on the mechanism proposed. The acid effect of reaction system, the structural effect of the complexes, the effect of surfactant micelles and the effect of temperature on the rate of BNPP hydrolysis catalyzed by the complexes have been discussed.     

Phosphate Diester Cleavage Promoted by the Metallomicelles of Ce(III) Complexes of Aza-Crown Ether with Different Numbers of Nitrogen Atoms

The design of artificial hydrolase has attracted extensive attention due to their scientific significance and potential application in the field of gene medicine and molecular biology. This work reports the catalytic activation of two aza-crown ether Ce(III) complexes and their metallomicelles as artificial hydrolase in bis(4-nitrophenyl) phosphate ester (BNPP) hydrolysis. The chemical composition of two complexes was determined by the fluorescence spectra and the mole ratio method for electronic absorption spectra. The bonding effect of BNPP and solubilizing effect of the complexes were proved by a method of fluorescence spectroscopy. The catalytic activity of different catalytic systems in BNPP hydrolysis was measured with UV-vis spectrophotometric method. These catalytic systems showed high catalytic activity for promoting BNPP hydrolysis at the almost physiological conditions. BNPP hydrolysis rate in these catalytic system is about 10⁷- to 10⁹-fold faster than that of the BNPP spontaneous hydrolysis in aqueous solution at the same conditions. The metallomicelle systems exhibited higher catalytic activity compared with the complex solution systems in BNPP hydrolysis, and hexadecyltrimethyl ammonium bromide micelle provides a useful catalytic environment for reaction. The acid effect of the catalytic system is ascribed to the formation of metal-bound hydroxide serving as a better kind of nucleophile.     

Copper(II) complexes of novel N-alkylated derivatives of cis,cis-1,3,5-triaminocyclohexane. 2. Metal-promoted phosphate diester hydrolysis

Aqueous copper(II) N,N',N' '-trimethyl-cis,cis-1,3,5-triaminocyclohexane (Cu(tach-Me(3))(2+)(aq)) promotes the hydrolysis of activated phosphate diesters in aqueous medium at pH 7.2. This complex is selective for cleavage of the phosphate diester sodium bis(p-nitrophenyl) phosphate (BNPP), the rate of hydrolysis of the monoester disodium p-nitrophenyl phosphate being 1000 times slower. The observed rate acceleration of BNPP hydrolysis is slightly greater than that observed for other Cu(II) complexes, such as [Cu([9]aneN(3))Cl(2)] ([9]aneN(3) identical with 1,4,7-triazacyclononane). The rate of hydrolysis is first-order in phosphate ester at low ester concentration and second-order in [Cu(tach-Me(3))](2+)(aq), suggesting the involvement of two metal complexes in the mechanism of substrate hydrolysis. The reaction exhibits saturation kinetics with respect to BNPP concentration according to a modified Michaelis-Menten mechanism: 2CuL + S <==> LCu-S-CuL --> 2CuL + products (K(M) = 12.3 +/- 1.8 mM(2), k(cat) = (4.0 +/- 0.4) x 10(-)(4) s(-1), 50 degrees C) where CuL (triple bond) [Cu(tach-Me(3))](2+), S (triple bond) BNPP, and LCu-S-CuL is a substrate-bridged dinuclear complex. EPR data indicate that the dicopper complex is formed only in the presence of BNPP; the active LCu-S-CuL intermediate species then slowly decays to products, regenerating monomeric CuL.     

Hydrolysis of BNPP Catalyzed by the Crowned Schiff Base Co(II) Complex Containing Benzoaza‐15‐Crown‐5 in Micellar Solution

It has been reported that three aza crowned Schiff base cobalt (II) complexes were synthesized and characterized, and the metallomicelle made up of the cobalt (II) complexes and surfactants(Brij35, CTAB, LSS), as mimic hydrolytic metalloenzyme, was used in catalytic hydrolysis of bis(4‐nitrophenyl) phosphate (BNPP). The analysis of specific absorption spectrums of the hydrolytic reaction systems indicated that key intermediates made up of BNPP and Co (II) complexes are formed in the reaction process of BNPP catalytic hydrolysis. The mechanism of BNPP catalytic hydrolysis proposed is based on the analytic result of specific absorption spectra. Based on the mechanism proposed, a kinetic mathematical model for the calculation of the kinetic parameter of BNPP catalytic hydrolysis has been established. The acid effect of reaction system, structure effect of the complexes, effect of temperature and effect of micelles on the rate of BNPP hydrolysis catalyzed by the complexes have been discussed.