Assuntos
Agricultura , Reguladores de Crescimento de Plantas , Transdução de Sinais/fisiologia , Congressos como Assunto , Reguladores de Crescimento de Plantas/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Reguladores de Crescimento de Plantas/fisiologia , Fenômenos Fisiológicos Vegetais/efeitos dos fármacos , Proteínas de Plantas/metabolismo , Proteínas de Plantas/fisiologia , Transdução de Sinais/efeitos dos fármacos , UcrâniaRESUMO
A series of thioureido derivatives of methylenebisphosphonic acid were synthesized by the reaction of aminomethylenebisphosphonic acid with the corresponding isothiocyanates, and their effect on the activity of alkaline phosphatases from bovine small intestine mucosa (BSIM) and human placenta was studied. It was found that (3-phenylthioureido)methylenebisphosphonate is approximately one order of magnitude more effective in inhibiting the activity of alkaline phosphatase from BSIM than the alkyl derivatives of thioureidomethylenebisphosphonic acid with methyl, ethyl, tert-butyl, or cyclohexyl substituents. The introduction of substituents into the benzene ring of (3-phenylthioureido)methylenebisphosphonate decreased the effect of the inhibitor on the activity of the enzyme. The affinity of (3-phenylureido)methylenebisphosphonate to the alkaline phosphatase of BSIM was also weaker as compared with the corresponding thioureidomethylenebisphosphonate. The insertion of thioureidobisphosphonates into the active site of alkaline phosphatase of human placenta by the method of molecular docking indicated that the methylenebisphosphonate residue and the substituted amino groups of the inhibitor are involved in the mechanisms of complex formation with the enzyme. It is supposed that the improvement of the inhibitory activity of (3-phenylthioureido)methylenebisphosphonate toward alkaline phosphatase of BSIM is due to the additional fixation of the phenyl substituent in the active site of the enzyme.
Assuntos
Fosfatase Alcalina/antagonistas & inibidores , Fosfatase Alcalina/química , Difosfonatos/química , Modelos Moleculares , Animais , Bovinos , Humanos , Intestino Delgado/enzimologia , Isotiocianatos/química , Placenta/enzimologiaRESUMO
Simple synthetic compounds of lauroyl-arginine ethyl ester (LAE) and 9-fluorenylmethoxycarbonyl-L-agrinine methyl ester (Fmoc-Arg-OMe) were studied for their inhibitory effect on the hydrolysis of chromogenic substrate Tos-Gly-Pro-Arg-pNA (Chromozym TH) by thrombin with K(i) for LAE 1.92 microM and 77 microM for Fmoc Arg-OMe. It was shown that LAE inhibits thrombin activity almost 20 times more strongly than trypsin (K(i) = 18.9 microM). At the same time LAE preserves the ability to be hydrolyzed by thrombin at pH 8.5 (k(cat) = 3.6 c(-1)) and trypsin (k(cat) = 56 c(-1)). It is suggested, that LAE ability to suppress growth of some microorganisms is conditioned to some extent by its ability to inhibit the activity of trypsin-like serine proteases, participating in the infection process.
Assuntos
Arginina/análogos & derivados , Fibrinolíticos , Fluorenos , Trombina/antagonistas & inibidores , Arginina/síntese química , Arginina/química , Arginina/farmacologia , Fibrinolíticos/síntese química , Fibrinolíticos/química , Fibrinolíticos/farmacologia , Fluorenos/síntese química , Fluorenos/química , Fluorenos/farmacologia , Hidrólise , Oligopeptídeos/química , Tripsina/químicaRESUMO
The inhibition of alkaline phosphatases by calix[4]arenes functionalysed at the macrocyclic upper rim by one or two methylenebisphosphonic acid fragments has been investigated. It is established, that calix[4]arene bismethylenebisphosphonic acid displayed stronger inhibition of alkaline phosphatase from bovine intestine mucosa than calix[4]arene methylenebisphosphonic acid. At the same time, the both inhibitors showed almost similar levels of inhibitory activities in respect of bovine kidney alkaline phosphatase or E. coli alkaline phosphatase. The tested compounds were docked computationally to the active site of the E. coli alkaline phosphatase. On the basis of results obtained the possible binding modes of inhibitors were analysed.
Assuntos
Fosfatase Alcalina/antagonistas & inibidores , Calixarenos/farmacologia , Inibidores Enzimáticos/farmacologia , Compostos Organofosforados/farmacologia , Fenóis/farmacologia , Animais , Sítios de Ligação , Calixarenos/química , Bovinos , Inibidores Enzimáticos/química , Escherichia coli/enzimologia , Mucosa Intestinal/enzimologia , Intestino Delgado/enzimologia , Rim/enzimologia , Modelos Moleculares , Estrutura Molecular , Fenóis/químicaRESUMO
The inhibiting effects of 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) and its 4-substituted derivatives in reactions of linoleyl acid or linoleyl alcohol oxidation catalyzed by potato tuber 5-lipoxygenase were investigated. Inhibiting properties of stable nitroxyl radicals in presence of lubrol and SDS were reduced at the transition from TEMPO to 4-hydroxy-TEMPO or 4-amino-TEMPO and increased at use of adamantane-1-carboxylic or 3-methyladamantane-1-carboxylic acid 1-oxyl-2,2,6,6-tetramethylpiperidine-4-yl esters. Enzyme activity at saturating concentrations of inhibitor was not suppressed completely, and decreased up to the certain level determined by the substrate nature. The dependence of partial inhibition efficiency on rotational correlation time of stable nitroxides in model micellar systems were analysed. It was supposed that 5-lipoxygenase inhibition includes the interaction of hydrophobic nitroxide with radical intermediate formed in enzymatic process.
Assuntos
Araquidonato 5-Lipoxigenase/química , Óxidos N-Cíclicos/química , Álcoois Graxos/química , Ácido Linoleico/química , Óxidos de Nitrogênio/química , Araquidonato 5-Lipoxigenase/isolamento & purificação , Catálise , Cinética , Estrutura Molecular , Oxirredução , Solanum tuberosum/químicaRESUMO
The linoleyl alcohol oxidation catalyzed by potato tuber 5-lipoxygenase was found to be efficiently inhibited by stable nitroxyl radicals: 1-oxyl-2,2,6,6-tetramethylpiperidin-4-yl 1-bicyclo[2,2,2]octane-1-carboxylate, 1-adamantylacetate, dodecanoate, and octadecanoate. The dependence of apparent IC50 values on the rotational correlation times of times of 4-hydroxy-1-oxyl-2,2,6,6-tetramethylpiperidine and its derivatives in model micellar systems was analyzed. The inhibition mechanism was proposed; it involves the interaction of hydrophobic nitroxyl radical with the intermediate radical enzyme-substrate complex.
Assuntos
Araquidonato 5-Lipoxigenase/química , Óxidos N-Cíclicos/química , Álcoois Graxos/química , Óxidos de Nitrogênio/química , Espectroscopia de Ressonância de Spin Eletrônica , Radicais Livres/química , Interações Hidrofóbicas e Hidrofílicas , Oxirredução , Solanum tuberosum/enzimologia , Marcadores de SpinRESUMO
Boc/Tos-L-Phe-L-Arg-Xaa tripeptides (where Xaa = L-Ala-OBut, L-Ala, or DL-AlaP (OC2H5)2) were synthesized by conventional methods of peptide synthesis in solution. Special features of their interaction with thrombin and trypsin were studied. Unlike trypsin, thrombin did not catalyze the hydrolysis of the L-Arg-L-AlaP-(OC2H5)2 bond. The Tos-L-Phe-L-Arg-DL-AlaP(OC2H5)2 peptide was the most active inhibitor of thrombin among all the compounds studied. The relationship between the structure and inhibitory action of the synthesized peptides is discussed. A part of this study was reported in the Augustusburg Conference of Advanced Science: Nucleic Acids--Targets and Tools, September 17-19, 2000, Germany.
Assuntos
Alanina/química , Antitrombinas/química , Antitrombinas/farmacologia , Peptídeos/química , Peptídeos/farmacologia , Fosfoaminoácidos/química , Antitrombinas/síntese química , Bioquímica/métodos , Hidrólise , Peptídeos/síntese química , Relação Estrutura-AtividadeRESUMO
A number of benzylalkylketones and benzylalkylcarbinols have been synthesized as non-hydrolizable substrate analogues of penicillin acylase (EC 3.5.1.11), and their affinity to the enzyme has been studied. The compounds with plane trigonal carbonyl group (ketones) were established to has bind to the enzyme 20-40 times more tightly than their tetrahedral counterparts with a hydroxyl function (carbinols). 4-Oxo-5-phenylpentanoic acid was found to be one of the most potent reversible competitive inhibitors of penicillin acylase with Ki-31 microM.
Assuntos
Compostos de Benzil/farmacologia , Escherichia coli/enzimologia , Cetonas/farmacologia , Penicilina Amidase/antagonistas & inibidores , Álcool Feniletílico/análogos & derivados , Alquilação , Hidrólise , Estrutura Molecular , Álcool Feniletílico/farmacologia , Relação Estrutura-Atividade , Especificidade por SubstratoRESUMO
Monoaryl of benzylphosphonic acid have been synthesized and studied as the inhibitors of penicillin acylase. These compounds were found to be effective and selective irreversible inhibitors of the enzyme. The kinetic parameters of enzyme inactivation are determined, and possible mechanism of the inhibition is discussed. These phosphonates should be useful as both penicillin acylase active site titrants and the tools for the enzyme function study. Benzylchloromethyl keton has been also prepared and it is an irreversible inhibitor of penicillin acylase.
Assuntos
Compostos de Benzil/farmacologia , Inibidores Enzimáticos/farmacologia , Escherichia coli/enzimologia , Organofosfonatos/farmacologia , Penicilina Amidase/antagonistas & inibidores , Fósforo/análise , Compostos de Benzil/química , Inibidores Enzimáticos/química , Ésteres , Hidrocarbonetos Clorados/farmacologia , Cinética , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
Artificial neural networks were used to analyze and predict the human immunodeficiency virus type 1 reverse transcriptase inhibitors. The training and control sets included 44 molecules (most of them are well-known substances such as AZT, dde, etc.). The activities of the molecules were taken from literature. Topological indices were calculated and used as molecular parameters. The four most informative parameters were chosen and applied to predict activities of both new and control molecules. We used a network pruning algorithm and network ensembles to obtain the final classifier. Increasing of neural network generalization of the new data was observed, when using the aforementioned methods. The prognosis of new molecules revealed one molecule as possibly very active. It was confirmed by further biological tests.
Assuntos
Desenho de Fármacos , Redes Neurais de Computação , Pirimidinas/química , Inibidores da Transcriptase Reversa , Algoritmos , Linhagem Celular , Transcriptase Reversa do HIV , HIV-1/efeitos dos fármacos , Humanos , Estrutura Molecular , Pirimidinas/farmacologia , Relação Estrutura-Atividade , Linfócitos T/microbiologia , Zidovudina/farmacologiaRESUMO
Phosphonic analogues of penicillin acylase substrates are found to be selective reversible competitive inhibitors of the enzyme from E. coli (EC 3.5.1.11). The mode of binding of the inhibitors to the enzyme and the influence of the stereoelectronic parameters of the phosphonic inhibitors on their affinity to the enzyme are discussed.
Assuntos
Escherichia coli/enzimologia , Organofosfonatos/farmacologia , Penicilina Amidase/antagonistas & inibidores , Fósforo/análise , Ligação Competitiva/fisiologia , Concentração de Íons de HidrogênioRESUMO
Phosphonate and phosphonoamidate derivatives of benzylphosphonic acids were synthesized as potential inhibitors of penicillin acylase (EC 3.5.1.11) proceeding from the concept of transition-state analogues. The compounds obtained are not the substrates of the enzyme and they are stable under conditions of enzyme activity testing.
Assuntos
Desenho de Fármacos , Organofosfonatos/química , Penicilina Amidase/antagonistas & inibidores , Fósforo/análise , Estabilidade de Medicamentos , Hidrólise , Estrutura Molecular , Organofosfonatos/síntese químicaRESUMO
3-Fluorotyrosine fluorescence is quenched effectively by phosphate ions not only by a dynamic but also by a static mechanism owing to H-bond complex formation in ground state. 3-Fluorotyrosine pKa values both in the ground and first excited state (8.3 and 4, respectively) are appreciably lower than those of tyrosine, thus promoting 3-fluorotyrosinate ion formation in the excited state. Additional emission owing to 3-fluorotyrosinate ion (near 350 nm) may be taken erroneously for tryptophan fluorescence.
RESUMO
Racemic N-Boc-4-difluoromethoxyphenylglycine was prepared by O-difluoromethylation of D-N-Boc-4-hydroxyphenylglycine under basic conditions, whereas the hexafluoropropylation reaction gives optically pure D-N-Boc-4-hexafluoropropoxyphenylglycine. D,L-4-difluoromethoxyphenylglycine was obtained by the action of TFA on the corresponding amino acid derivative.
RESUMO
It is found that yeast pyruvate decarboxylase is inhibited by alkyl phosphates. Inhibition is competitive with respect to a substrate. The inhibition constants with n-butyl and n-heptyl esters of phosphoric acid are the values of the same order of magnitude. With an increase in the length of the alkyl phosphates hydrocarbon chain from 7 to 10 carbon atoms inhibition constants change drastically. For n-heptyl phosphate and n-decyl phosphate values KI are equal to 1.6 x 10(-4) M and 1.7 x 10(-6) M, respectively. A further increase in the number of carbon atoms in the alkyl substituent of phosphoric acid ester induces no reduction of the inhibition constant. Multiple-inhibitor experiments of pyruvate decarboxylase show that inorganic phosphate and n-decyl ester of phosphoric acid are mutually exclusive. It is suggested that the inhibition mechanism with alkyl phosphates includes the competition of the phosphoric acid residue with alpha-ketocarboxyl group of pyruvate as well as the interaction between a hydrocarbon radical and hydrophobic parts on the enzyme surface, one of them being outside the substrate binding site.
Assuntos
Fosfatos/farmacologia , Piruvato Descarboxilase/antagonistas & inibidores , Saccharomyces cerevisiae/enzimologia , Alquilação , Ligação Competitiva/fisiologia , Especificidade por SubstratoRESUMO
We found that (R,S)-2-hydroxy-2-trifluoromethyl-trans-n-octadec-4-enoic acid (HTFOA) is a powerful activator of 5-lipoxygenase from potato tubers. The degree of activation of the enzyme is proportional to the HTFOA concentration and is a maximum at about 0.1 mM independently of initial substrate concentration (25 microM or 0.1 mM). At greater concentrations of HTFOA, enzyme inhibition takes place. Enzyme activation is inversely proportional to the substrate (linoleic acid) concentration. The results may be explained by assuming that a regulatory center exists in the enzyme molecule, which shows affinity to both substances: activator and linoleic acid.
Assuntos
Araquidonato 5-Lipoxigenase/metabolismo , Ácidos Graxos Monoinsaturados/farmacologia , Hidroxiácidos/farmacologia , Solanum tuberosum/enzimologia , Ativação Enzimática , Ácidos Graxos Monoinsaturados/química , Hidroxiácidos/química , Cinética , Ácido Linoleico , Ácidos Linoleicos/química , OxirreduçãoAssuntos
Araquidonato 5-Lipoxigenase/efeitos dos fármacos , Ácidos Graxos/farmacologia , Adsorção , Araquidonato 5-Lipoxigenase/metabolismo , Ativação Enzimática/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Micelas , Oxirredução/efeitos dos fármacos , Relação Estrutura-AtividadeRESUMO
The chromatographic behaviour of some alpha-trifluoromethyl-alpha-aminoacids on L-proline- and L-hydroxyproline sorbents was studied. The retention and selectivity parameters of the separation of amino acid enantiomers on the sorbents were determined. The introduction of a CF3 [corrected] group led to an increased selectivity in the separation of amino acid enantiomers on a proline sorbent and to a decreased selectivity on a hydroxyproline sorbent.
Assuntos
Aminoácidos/isolamento & purificação , Adsorção , Cromatografia Líquida de Alta Pressão , Cromatografia por Troca Iônica , Cobre , Sulfato de Cobre , Hidroxiprolina/isolamento & purificação , Cinética , Ligantes , Prolina/isolamento & purificaçãoRESUMO
Penicillin acylase from E. coli (EC 3.5.1.11) was found to hydrolyze N-phenylacetylated 1-aminoethylphosphonic acid and its esters. The enzyme preferentially converts the R-form of the substrates: the ratios of the bimolecular rate constants of penicillin acylasecatalyzed hydrolysis of R- and S-forms of 1-(N-phenylacetamino)-ethylphosphonic acid and its dimethyl- and diisopropyl-esters are 58000, 2300, 1800; these derivatives were shown to have the greatest values of the catalytic constants for enzymatic hydrolysis of all known substrates for penicillin acylase: 237, 148 and 134 s-1; the corresponding Km values are 3.7 10(-5), 6.8 10(-4) and 6.2 10(-4) M at pH 7.0. The kinetics of enzymatic hydrolysis of 1-(N-phenylacetamino)-ethylphosphonic acid was investigated up to high degrees of conversion. The inhibition of penicillin acylase by high concentrations of the R-form of the substrate (with substrate inhibition constant of 0.07 M) and competitive inhibition by the reaction product, phenylacetic acid (Ki = 3.5 10(-5) M), was observed.