Model Studies of TTQ-Containing Amine Dehydrogenases

The reactions of a TTQ model compound [1, 3-methyl-4-(3'-methylindol-2'-yl)indole-6,7-dione] with several amines have been investigated in organic media to obtain mechanistic information on the action of quinoprotein methylamine and aromatic amine dehydrogenases. It has been found that compound 1 acts as an efficient catalyst for the autorecycling oxidation of benzylamine by molecular oxygen in CH(3)OH. In order to evaluate the oxidation mechanism of amines by 1, the product analyses and kinetic studies have been carried out under anaerobic conditions. In the first stage of the reaction of 1 with amines, 1 is converted into an iminoquinone-type adduct (so-called substrateimine), which was isolated and characterized by using cyclopropylamine as a substrate. The observed NOE of the isolated product indicates clearly that the addition position of the amine is C-6 of the quinone. The molecular orbital calculations suggest that the thermodynamic stability of the carbinolamine intermediate is a major factor to determine such regioselectivity; the C-6 carbinolamine is more stable than the C-7 counterpart by 2.9 kcal/mol. The reactivity of several primary amines and the electronic effect of the p-substituents of benzylamine derivatives in the iminoquinone formation suggest that the addition step of the amine to the quinone is rate-determining. When amines having an acidic alpha-proton such as benzylamine derivatives are employed as substrates, formation of the iminoquinone adduct was followed by rearrangement to the productimine. The kinetic analysis has revealed that this rearrangement consists of noncatalyzed and general base-catalyzed processes. Large kinetic isotope effects of 7.8 and 9.2 were observed for both the noncatalyzed and general base-catalyzed processes, respectively, since these steps involve a proton abstraction from the alpha-position of the substrate. In the reaction with benzhydrylamine, the product imine was isolated quantitatively and well characterized by several spectroscopic data. In the case of benzylamine, the product imine is further converted into the aminophenol derivative by the imine exchange reaction with excess benzylamine. These results indicate clearly that the amine oxidation by compound 1 proceeds via a transamination mechanism as suggested for the enzymatic oxidation of amines by TTQ cofactor.     

Cofactor diversity in biological oxidations: implications and applications

Until recently, it was generally believed that enzymatic oxidation and reduction requires the participation of either a nicotinamide (NAD(P)+) or a flavin (FAD, FMN), in agreement with the existence of NAD(P)/H-dependent dehydrogenases/reductases and flavoprotein dehydrogenases/reductases/oxidases. However, during the past 20 years, the unraveling of the enzymology of the oxidation and reduction of C1-compounds by bacteria has led to the discovery of many new redox cofactors, some of them discussed here as they have a wider physiological significance than just enabling enzymatic C1-conversions to occur. A good example is the quinone cofactors, encompassing PQQ (2,7,9-tricarboxy-1H-pyrrolo[2,3-f]-quinoline-4,5-dione), TTQ (tryptophyl tryptophanquinone), TPQ (topaquinone), LTQ (lysyl topaquinone), and several others whose structures have still to be elucidated. Another example is mycothiol (1-O-(2'-[N-acetyl-L-cysteinyl]amido-2'-deoxy-alpha-D-glucopyranosyl)-D-myo-inosoitol), the counterpart of glutathione, once thought to be a universal coenzyme. Because these novel cofactors assist in reactions that can also be catalyzed by already known enzyme "classic cofactor" combinations, and first indications suggest that the chemistry of the reactions is not unique, one may wonder about the evolutionary background for this cofactor diversity. However, as will be illustrated by examples, from a practical point of view the diversity is beneficial, as it has increased the arsenal of enzymes suitable for application.     

Molecular Orbital Calculation and Spectroscopic Study of the Photochemical Generation of Bis(2,2'-bipyridine)rhodium(I) from Bis(2,2'-bipyridine)(oxalato)rhodium(III)

A planar complex, [Rh(bpy)(2)](+) (bpy = 2,2'-bipyridine), was obtained from [Rh(ox)(bpy)(2)](+) (ox = oxalato) by photoirradiation. A rate constant k for the photoreaction was evaluated as 1 x 10(8) s(-1) in simple first-order kinetics, whereas a ligand dissociation, a reorganization of the coordinated bpy, and a two-electron transfer were involved in the reaction. The process of the Rh(I) complex generation was investigated in terms of a discrete variational (DV)-Xalpha molecular orbital calculation on [Rh(ox)(HN=CHCH=NH)(2)](+) instead of [Rh(ox)(bpy)(2)](+). From the calculation, using the transition-state method, it was predicted that a transition of the ox pi orbital to the metal 4d(z)()2 orbital caused the ligand dissociation and the reorganization of the coordinated bpy occurred in the ox pi to Rh 4d(x)()2(-y)2 excited state stabilized by the ox elimination. Upon release of the ligand and a change from octahedral to square-planar geometry, the electron density on the metal increased and the Rh 4d orbital acquired a d(8) electronic configuration.     

Synthesis,structure, and oxidation of 3,4-dihydro-1,2,5-benzotrithiepins

Stable benzene-fused polysulfide compounds, 3,4-dihydro-1,2,5-benzotrithiepins ( 1a-c ), have been prepared, and the structure of 1a has been determined by X-ray crystallographic analysis. While the electrophilic oxidation of compounds 1 with m-chloroperbenzoic acid gave the corresponding 3,4-dihydro-1,2,5-benzotrithiepin 5-oxides ( 2 ) in moderate yields, the oxidation of 1 with N-bromosuccinimide afforded a mixture of 5-oxides 2 , unexpected, inseparable 3,4-dihydro-1,2,5-benzotrithiepin 2,2-dioxides ( 3 ), and 3,4-dihydro-1,2,5-benzotrithiepin 1,1-dioxides ( 4 ). Semiempirical PM3 calculations were carried out, and the computed HOMO of 1a suggested a significant favoring of electrophilic reactions at the sulfur atom at the 5-position. The treatment of 5-oxides 2 with acetyl bromide or oxalyl dibromide as halogenating reagents gave 2,2-dioxides 3 and 1,1-dioxides 4 , suggesting that an intramolecular halogen transfer from the 5-position (sulfide moiety) to the 1- and 2-positions (disulfide moiety) took place in the reactions.     

Chemoselective C-4 aerobic oxidation of catechin derivatives catalyzed by the Trametes villosa laccase/1-hydroxybenzotriazole system: synthetic and mechanistic aspects

Catechin derivatives were oxidized in air in the presence of the Trametes villosa laccase/1-hydroxybenzotriazole (HBT) system in buffered water/1,4-dioxane as reaction medium. The oxidation products, flavan-3,4-diols and the corresponding C-4 ketones, are bioactive compounds and useful intermediates for the hemisynthesis of proanthocyanidins, plant polyphenols which provide beneficial health properties for humans. Determinations of oxidation potentials excluded that catechin derivatives could be directly oxidized by laccase Cu(II), while it resulted in the H-abstraction from benzylic positions being promptly promoted by the enzyme in the presence of the mediator HBT, the parent species producing in situ the reactive intermediate benzotriazole-N-oxyl (BTNO) radical. A remarkable and unexpected result for the laccase/HBT oxidative system has been the chemoselective insertion of the oxygen atom into the C-4-H bond of catechin derivatives. Mechanistic aspects of the oxidation reaction have been investigated in detail for the first time in order to corroborate these results. Since the collected experimental findings could not alone provide information useful to clarify the origin of the observed chemoselectivity, these data were expressly supplemented with information derived by suitable molecular modeling investigations. The integrated evaluation of the dissociation energies of the C-H bonds calculated both by semiempirical and DFT methods and the differential activation energies of the process estimated by a molecular modeling approach suggested that the observed selective oxidation at the C-4 carbon has a kinetic origin.     

Formation of pyrrole derivatives through aminolysis of 2-azetidinone derivatives

A mixture of 3-(α,β-epoxyisopropyl)-1-phenyl-2-azetidinone and benzylamine was heated in a sealed tube at 120–130° yielding 4-anilinomethyl-1-benzyl-3-hydroxy-3-methyl-2-pyrrolidinone as a mixture of diastereoisomers. By this method, 4-anilinomethyl-3-hydroxy-3-methyl-1-phenyl-2-pyrrolidinone and 4-anilinomethyl-3-hydroxy-3-methyl-1-(3,4-dimethoxyphenethyl)-2-pyrrolidinone were obtained by using aniline and 3,4-dimethoxyphenethylamine, respectively, instead of benzylamine. The reaction of 4-formyl-1-phenyl-2-azetidinone with 3,4-dimethoxyphenethylamine afforded 4-anilino-1-(3,4-dimethoxyphenethyl)-2,3-dihydro-2-oxopyrrole. In a similar fashion, the 1-n-butyl and 1-isobutyl analogues were obtained by the use of n-butylamine and isobutylamine, respectively, instead of 3,4-dimethoxyphenethylamine.     

Synthesis, structure, and characterisation of a new phenolato-bridged manganese complex [Mn2(mL)2]2+: chemical and electrochemical access to a new mono-mu-oxo dimanganese core unit

The dinuclear phenolato-bridged complex [(mL)Mn(II)Mn(II)(mL)](ClO(4))(2) (1(ClO(4))(2)) has been obtained with the new [N(4)O] pentadentate ligand mL(-) (mLH=N,N'-bis-(2-pyridylmethyl)-N-(2-hydroxybenzyl)-N'-methyl-ethane-1,2-diamine) and has been characterised by X-ray crystallography. X- and Q-band EPR spectra were recorded and their variation with temperature was examined. All spectra exhibit features extending over 0-800 mT at the X band and over 100-1450 mT at the Q band, features that are usually observed for dinuclear Mn(II) complexes. Cyclic voltammetry of 1 exhibits two irreversible oxidation waves at E(1)(p)=0.89 V and E(2)(p)=1.02 V, accompanied on the reverse scan by an ill-defined cathodic wave at E(1')(p)=0.56 V (all measured versus the saturated calomel electrode (SCE)). Upon chemical oxidation with tBuOOH (10 equiv) at 20 degrees C, 1 is transformed into the mono-mu-oxo species [(mL)Mn(III)-(mu-O)-Mn(III)(mL)](2+) (2), which eventually partially evolves into the di-mu-oxo species [(mL)Mn(III)-(mu-O)(2)-Mn(IV)(mL)](n+) (3) in which one of the aromatic rings of the ligand is decoordinated. The UV/Vis spectrum of 2 displays a large absorption band at 507 nm, which is attributed to a phenolate-->Mn(III) charge-transfer transition. The cyclovoltammogram of 2 exhibits two reversible oxidation waves, at 0.65 and 1.16 V versus the SCE, corresponding to the Mn(III)Mn(III)/Mn(III)Mn(IV) and Mn(III)Mn(IV)/Mn(IV)Mn(IV) oxidation processes, respectively. The one-electron electrochemical oxidation of 2 leads to the mono-mu-oxo mixed-valent species [(mL)Mn(III)-(mu-O)-Mn(IV)(mL)](3+) (2 ox). The UV/Vis spectrum of 2 ox exhibits one large band at 643 nm, which is attributed to the phenolate-->Mn(IV) charge-transfer transition. 2 ox can also be obtained by the direct electrochemical oxidation of 1 in the presence of an external base. The 2 ox and 3 species exhibit a 16-line EPR signal with first peak to last trough widths of 125 and 111 mT, respectively. Both spectra have been simulated by using colinear rhombic Mn-hyperfine tensors. Mechanisms for the chemical formation of 2 and the electrochemical oxidation of 1 into 2 ox are proposed.     

A novel one-pot oxidation polymerization of dithiols obtained from bifunctional five-membered cyclic dithiocarbonates with amines

One-pot oxidation polymerization of dithiols, obtained from bifunctional five-membered cyclic dithiocarbonates ( 4a and 4b ) with two equivalents of amines, was studied. The monomers 4a and 4b were synthesized by the reactions of bisphenol A diglycidyl ether and neopentyl glycol diglycidyl ether with carbon disulfide, respectively. Polydisulfides with M̄_ns 4600–20,200 were obtained quantitatively in the oxidation polymerization of the dithiols obtained in situ by the reactions of 4a with benzylamine, n-butylamine, and piperidine. On the other hand, dithiols obtained from 4b with benzylamine, afforded cyclic disulfides as well as the polydisulfide under similar conditions. © 1998 John Wiley & Sons, Inc. J Polym Sci A: Polym Chem 36: 79–84, 1998     

Model studies on p-hydroxybenzoate hydroxylase. The catalytic role of Arg-214 and Tyr-201 in the hydroxylation step

A model C-(4a)-flavinhydroperoxide (FlHOOH) is described that contains the tricyclic isoalloxazine moiety, the C-4a-hydroperoxide functionality, and a beta-hydroxyethyl group to model the effect of the 2'-OH group of the ribityl side chain of native FADHOOH. The electronic structures of this reduced flavin (H(3)()Fl(red)()), its N1 anion (H(2)()Fl(red)()(-)()), oxidized flavin (HFl(ox)()), and FlHOOH have been fully optimized at the B3LYP/ 6-31+G(d,p) level of theory. This model C-4a-flavinhydroperoxide is used to describe the transition state for the key step in the paradigm aromatic hydroxylase, p-hydroxybenzoate hydroxylase (PHBH): the oxidation of p-hydroxybenzoate (p-OHB). The Tyrosine-201 residue in PHBH is modeled by phenol, and Arginine-214 is modeled by guanidine. Electrophilic aromatic substitution proceeds by an S(N)2-like attack of the aromatic sextet of p-OHB phenolate anion on the distal oxygen of FlHOOH 3. The transition structure for oxygen atom transfer is fully optimized [B3LYP/6-31+G(d,p)] and has a classical activation barrier of 24.9 kcal/mol. These data suggest that the role of the Tyr-201 is to orient the p-OHB substrate and to properly align it for the oxygen transfer step. Although the negatively charged phenolate oxygen does activate the C-3 carbon of p-OHB phenolate anion toward oxidation relative to ortho oxidation of the carboxylate anion, it appears that H-bonding the Tyr-201 residue to this phenolic oxygen stabilizes both the ground state (GS) and the transition state (TS) approximately equally and therefore plays only a minor role, if any, in lowering the activation barrier. Complexation of p-OHB with guanidine has only a modest effect upon the oxidation barriers. When the complex is in the form of a salt-bridge (10a), the barrier is only slightly reduced. When the TSs are placed in THF solvent (COSMO) with full geometry optimization, salt-bridge TS-A is slightly favored (DeltaDeltaE() = 2.3 kcal/mol).     

Lignin peroxidase-catalyzed oxidation of nonphenolic trimeric lignin model compounds: fragmentation reactions in the intermediate radical cations

The H(2)O(2)-promoted oxidations of the two nonphenolic beta-O-aryl lignin model trimers 1 and 2, catalyzed by lignin peroxidase (LiP) at pH = 3.5, have been studied. The results have been compared with those obtained in the oxidation of 1 and 2 with the genuine one-electron oxidant potassium 12-tungstocobalt(III)ate. These models present a different substitution pattern of the three aromatic rings, and by one-electron oxidation, they form radical cations with the positive charge, which is localized in the dialkoxylated ring as also evidenced by a pulse radiolysis study. Both the oxidations with the enzymatic and with the chemical systems lead to the formation of products deriving from the cleavage of C-C and C-H bonds in a beta position with respect to the radical cation with the charge residing in the dialkoxylated ring (3,4-dimethoxybenzaldehyde (5) and a trimeric ketone 6 in the oxidation of 1 and a dimeric aldehyde 8 and a trimeric ketone 9 in the oxidation of 2). These products are accompanied by a dimeric aldehyde 7 in the oxidation of 1 and 4-methoxybenzaldehyde (10) in the oxidation of 2. The unexpected formation of these two products has been explained by suggesting that 1.+ and 2.+ can also undergo an intramolecular electron transfer leading to the radical cations 1a.+ and 2a.+ with the charge residing in a monoalkoxylated ring. The fast cleavage of a C-C bond beta to this ring, leading to 7 from 1.+ and to 10 from 2.+, is the driving force of the endoergonic electron transfer. A kinetic steady-state investigation of the LiP-catalyzed oxidation of the trimer 2, the dimeric model 1-(3,4-dimethoxyphenyl)-2-phenoxy-1-ethanol (4), and 3,4-dimethoxybenzyl alcohol (3) has indicated that the turnover number (k(cat)) and the affinity for the enzyme decrease significantly by increasing the size of the model compound. In contrast, the three substrates exhibited a very similar reactivity toward a chemical oxidant [Co(III)W]. This suggests a size-dependent interaction of the enzyme with the substrate which may influence the efficiency of the electron transfer.     

Novel stereochemistry, reactivity, and stability of an arsenic heterocycle in a metal-promoted asymmetric cycloaddition reaction

The organopalladium complex containing ortho-metalated (S)-[1-(dimethylamino)ethyl]phenylene as the chiral auxiliary has been used as the chiral template to promote the asymmetric cycloaddition reaction between diphenylvinylphosphine and 3,4-dimethyl-1-phenylarsole. A diphenylphosphino-substituted asymmetrical heterobidentate arsanorbornene (As-P) ligand was obtained stereoselectively on the chiral palladium template in moderate yield. The chiral benzylamine auxiliary could be removed chemoselectively from the template by treatment with HCl to produce the neutral complex [(As-P)PdCl2]. In contrast to their reported P-P analogue, the arsenic donor in the dichloro complex could be eliminated stereospecifically under mild reaction conditions to generate the corresponding 1-(diphenylphosphino)-3,4-dimethyl-2,4-cyclohexadiene, which remained as a bidentate ligand at the PdCl2 unit via phosphorus and the eta2-C4-C5 double bond. The arsenic-elimination process was found to be influenced by the halo ligand in [(As-P)PdX2]. A similar process was observed with the analogous dibromo complex, but the corresponding diiodo species did not show similar reactivity. All of the novel As-Pd complexes have been characterized by X-ray crystallography.     

1,4-dioxobenzene compounds of gallium: reversible binding of pyridines to [[((t)Bu)(2)Ga](2)(mu-OC(6)H(4)O)](n) in the solid state

The gallium aryloxide polymer, [[((t)Bu)(2)Ga](2)(mu-OC(6)H(4)O)](n)(1) is synthesized by the addition of Ga((t)()Bu)(3) with hydroquinone in a noncoordinating solvent, and reacts with pyridines to yield the yellow compound [((t)()Bu)(2)Ga(L)](2)(mu-OC(6)H(4)O) [L = py (2), 4-Mepy (3), and 3,5-Me(2)py (4)] via cleavage of the Ga(2)O(2) dimeric core. The analogous formation of Ga((t)()Bu)(2)(OPh)(py) (5) occurs by dissolution of [((t)Bu)(2)Ga(mu-OPh)](2) in pyridine. In solution, 2-4 undergo dissociation of one of the pyridine ligands to yield [((t)()Bu)(2)Ga(L)(mu-OC(6)H(4)O)Ga((t)Bu)(2)](2), for which the DeltaH and DeltaS have been determined. Thermolysis of compounds 2-4 in the solid-state results in the loss of the Lewis base and the formation of 1. The reaction of 1 or [((t)Bu)(2)Ga(mu-OPh)](2) with the vapor of the appropriate ligand results in the solid state formation of 2-4 or 5, respectively. The deltaH and deltaS for both ligand dissociation and association for the solid-vapor reactions have been determined. The interconversion of 1 into 2-4, as well as [((t)Bu)(2)Ga(mu-OPh)](2) into 5, and their reverse reactions, have been followed by (13)C CPMAS NMR spectroscopy, TG/DTA, SEM, EDX, and powder XRD. Insight into this solid-state polycondensation polymerization reaction may be gained from the single-crystal X-ray crystallographic packing diagrams of 2-5. The crystal packing for compounds 2, 3, and 5 involve a head-to-head arrangement that is maintained through repeated ligand dissociation and association cycles. In contrast, when compound 4 is crystallized from solution a head-to-tail packing arrangement is formed, but during reintroduction of 3,5-Me(2)py in the solid state-vapor reaction of compound 1, a head-to-head polymorph is postulated to account for the alteration in the deltaH of subsequent ligand dissociation reactions. Thus, the deltaH for the condensation polymerization reaction is dependent on the crystal packing; however, the subsequent reversibility of the reaction is dependent on the polymorph.     

Enzymes do what is expected (chalcone isomerase versus chorismate mutase)

Madicago sativa chalcone isomerase (CI) catalyzes the isomerization of chalcone to flavanone, whereas E. coli chorismate mutase (CM) catalyzes the pericyclic rearrangement of chorismate to prephenate. Covalent intermediates are not formed in either of the enzyme-catalyzed reactions, K(M) and k(cat) are virtually the same for both enzymes, and the rate constants (k(o)) for the noncatalyzed reactions in water are also the same. This kinetic identity of both the enzymatic and the nonenzymatic reactions is not shared by a similarity in driving forces. The efficiency (DeltaG(o)() - DeltaG(cat)()) for the CI mechanism involves transition-state stabilization through general-acid catalysis and freeing of three water molecules trapped in the E.S species. The contribution to lowering DeltaG(cat)() by an increase in near attack conformer (NAC) formation in E.S as compared to S in water is not so important. In the CM reaction, the standard free energy for NAC formation in water is 8.4 kcal/mol as compared to 0.6 kcal/mol in E.S. Because the value of (DeltaG(o)() - DeltaG(cat)()) is 9 kcal/mol, the greater percentage of NACs accounts for approximately 90% of the kinetic advantage of the CM reaction. There is no discernible transition-state stabilization in the CM reaction. These results are discussed. In anthropomorphic terms, each enzyme has had to do what it must to have a biologically relevant rate of reaction.     

Octahedral non-heme non-oxo Fe(IV) species stabilized by a redox-innocent N-methylated cyclam-acetate ligand

The ligand 1,4,8-tri-N-methyl-1,4,8,11-tetraazacyclotetradecane-11-acetic acid (Me3cyclam-acetic acid) has been synthesized by Eschweiler-Clarke methylation of cyclam-acetic acid, and the iron(III) complex [(Me3cyclam-acetate)FeN3]PF6, 1, has been synthesized, which has been found to have significantly different properties than its unmethylated analogue, [(cyclam-acetate)FeN3]PF6, 2. Whereas the iron ion in 2 is low spin with S = 1/2, 1 is found to be high spin at temperatures above 100 K, though low-spin species are observed at lower temperatures, indicating a spin crossover phenomenon. The iron(II) species 1red is electrochemically more accessible than 2red since the Fe2+/3+ redox wave in 1 appears approximately 350 mV more positive than the corresponding wave in 2. Also, 1 displays a reversible Fe3+/4+ redox wave, which is irreversible in 2, denoting that the Fe(IV) species 1ox is kinetically stable. 1red and 1ox have been generated electrochemically in solution and studied spectroscopically. M?ssbauer spectroscopy has confirmed that, in both reduction and oxidation, iron is the redox center, that 1red is high spin (S = 2), and that 1ox is low spin (S = 1), in contrast to 2red which is low spin and 2ox which could not be isolated.     

The Different,but Interesting Behaviors of Benzyl Systems in the Willgerodt-Kindler Reaction under Solvent-Free Conditions

On the benzyl system, bearing various functional groups, have been carried out the Willgerodt-Kindler reaction to obtain thiobenzmorpholide ( 1 ). The reactions, under solvent-free conditions, were performed in both classical (reflux, room temperature) and nonclassical (microwave) conditions to attempt our elucidation of the reactions pathways. Unlike benzylamine and benzyl mercaptan, benzyl halides give poor result due to the type of amine. The experimental results suggest that the proposed reaction pathway involves the oxidation coupling of benzylic substrates, followed by a thiolation step and an attack of the amine on the thiolated product to give ( 1 ).     

Electrochemistry of 6-methyl-5,6,7,8-tetrahydropterin

The electrochemical oxidation of 6-methyl-5,6,7,8-tetrahydropterin (6-MTHP), the most effective of the synthetic aromatic amino acid hydroxylase pseudo cofactors, has been studied in aqueous solution over a wide pH range at a pyrolytic graphite electrode. The first electrooxidation of 6-WTHP occurs by a quasi-reversible 2e-2H⁺ reaction giving an unstable quinonoid-dihydropterin. The latter undergoes a first order chemical follow-up reaction yielding 6-methyl-7,8-dihydropterin (6-MDHP). At pH values ?5.6 6-MDHP forms an equilibrium mixture of a covalently hydrated species and non-hydrated species. The covalently hydrated form of 6-MDHP is electrooxidized in a 2e-2H⁺ quasi-reversible reaction to another unstable quinonoid that appears to undergo a two-step rearrangement to 6-methylpterin. Non-hydrated 6-MDHP is electrooxidized at the most positive potential in an irreversible 2e-2H⁺ reaction giving 6-methylpterin.     

Single-site beta-diiminate zinc catalysts for the ring-opening polymerization of beta-butyrolactone and beta-valerolactone to poly(3-hydroxyalkanoates)

Polymerization of beta-butyrolactone (BBL) and beta-valerolactone (BVL) using the zinc alkoxide initiator (BDI-1)ZnO(i)()Pr [(BDI-1) = 2-((2,6-diisopropylphenyl)amido)-4-((2,6-diisopropylphenyl)imino)-2-pentene] proceeds very rapidly under mild conditions to produce poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxyvalerate) (PHV), respectively. For the monomer-to-initiator ratio 200:1, PHB number-average molecular weights (M(n)) are proportional to conversion throughout the reaction and polydispersity indices (PDIs) are narrow, consistent with a living polymerization. Higher monomer-to-initiator ratios can be used to achieve high molecular weight PHB (M(n) > 100 000). End-group analysis verifies that the polymerization of BBL follows a coordination-insertion mechanism, where complexes of the form (BDI-1)ZnOCH(Me)CH(2)CO(2)R are the active species. Variable temperature NMR experiments show that (BDI-1)ZnO(i)()Pr is monomeric in benzene-d(6) solution. In contrast, (BDI-2)ZnO(i)()Pr [(BDI-2) = 2-((2,6-diethylphenyl)amido)-4-((2,6-diethylphenyl)imino)-2-pentene] is a poor initiator at room temperature because it prefers to form a bis-mu-isopropoxide dimer in solution. According to kinetic studies, propagation by (BDI-1)ZnO(i)()Pr is first order in both monomer as well as (BDI-1)ZnO(i)()Pr concentration. These results lead us to propose a monometallic active species. Several results suggest that elimination side reactions are slowly catalyzed by zinc alkoxides, leading to degradation of the polymer.     

Electrochemical oxidation of 4,5,6,7-tetrahydro-1H-pyrazolo[3,4-d] pyrimidine-4,6-dione (oxipurinol) at the pyrolytic graphite electrode

The electrochemical oxidation of 4,5,6,7-tetrahydro-1H-pyrazolo[3,4-d] pyrimidine-4,6-dione (oxipurinol) at the pyrolytic graphite electrode (PGE) has been studied. Oxipurinol exhibits up to three voltammetric oxidation peaks at the PGE between pH 1–12. The first pH-dependent peak (peak I_a) is proposed to be an initial, irreversible 2e-2H⁺ reaction to give 5,6-dihydro-4H-pyrazolo[3,4-d] pyrimidine-4,6-dione. This primary product further reacts by two routes. The major route, accounting for ca. 90% of the latter compound, involves a Michael addition of water followed by further electrochemical oxidation and hydrolysis to give 5,6-dihydro-5,6-dihydroxy-5-carboxy-6-diazenouracil. The minor route involves further electrochemical oxidation of 5,6-dihydro-4H-pyrazolo[3,4-d]-pyrimidine-4,6-dione in a 2e-2H⁺ reaction to give 4,5,6,7-tetrahydro-3H-pyrazolo[3,4-d]-pyrimidine-3,4,6-trione.Decomposition and, generally, additional electrochemical reactions of 5,6-dihydro-5,6-dihydroxy-5-carboxy-6-diazenouracil result in the formation of alloxan, parabanic acid, 6-diazo-isobarbituric acid and 5′-hydroxy-5-carboxy-6,6′-azouracil. The two latter compounds have never previously been reported. Decomposition of 4,5,6,7-tetrahydro-3H-pyrazolo[3,4-d]pyrimidine-3,4,6-trione results in formation of uracil-5-carboxylic acid.Detailed reaction schemes have been proposed to explain the observed electrochemistry and the formation of the observed products.     

Spectrofluorimetric determination of 5-hydroxyindoles with benzylamine or 3,4-dimethoxybenzylamine as a selective fluorogenic reagent.

A spectrofluorimetric method has been developed for the sensitive and selective determination of 5-hydroxyindoles; the method is based on the reaction of 5-hydroxyindoles in a weakly alkaline solution (pH 9.0) with aromatic methylamines in the presence of potassium hexacyanoferrate(III) and dimethyl sulphoxide; the compounds produced fluoresce intensely in an alkaline solution (pH 11-12). Of the eight aromatic methylamines tested, benzylamine and 3,4-dimethoxybenzylamine were found to be the most favourable fluorogenic reagents in terms of sensitivity and reactivity. The methods with benzylamine and 3,4-dimethoxybenzylamine permit the determination of 5-hydroxyindoles at concentrations as low as 22-72 pmol ml-1 and 1.0-2.4 nmol ml-1, respectively.     

2－氯／2－甲磺酰基－5－（2－苯基－4－喹啉基）－1，3，4－噁二唑化合物的合成及其亲核取代的研究

借处理２－羟基－５－（２－苯基－４－喹啉基）－１，３，二唑同ＰＣｌ＿５／ＰＯＣｌ＿３之间的反应合成了２－氯－５－（２－苯基－４－喹啉基）－１，３，４－二唑（３）和通过２－基－５－（２－苯基－４－喹啉基）－１，３，４－二唑的甲基化，然后氧化制得２－甲磺酰基－５－（２－苯基－４－喹啉基）－１，３，４－二唑（６）．并分别研究了３和６同胺、叠氮及肼的反应，得到２，５－二取代的二唑新衍生物．初步观察了部分化合物的抗菌活性．