[Inhibition of alkaline phosphatase by thioureido derivatives of methylenebisphosphonic acid].

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.

HIV-1 reverse transcriptase inhibitor design using artificial neural networks.

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.

[Reverse phase high performance liquid chromatography in the synthesis of leukotrienes A4 and C4]. / Obrashchenno-fazovaia vysokoéffektivnaia zhidkostnaia khromatografiia v sinteze leikotrienov A4 i C4.

The chromatographic (RP HPLC) behaviour of leukotriene C4, its methyl ester, leukotriene A4 methyl ester and some chemicals involved in their synthesis have been investigated. Optimal conditions of separation were determined for the gradient and isocratic HPLC. Parameters of the interaction of the substances with hydrophobic surface are discussed in terms of solvophobic theory.

[Synthesis and antithrombin activity of phosphoalanine-containing peptides]. / Sintez i uzuchenie antitrombinovoi aktivnosti pepidov, soderzhashchikh fosfoalanin.

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.

[Activation of 5-lipoxygenase by lipophilic n-alkyl-containing acids--an allosteric process]. / Aktivatsiia 5-lipoksigenazy lipofil'nymi n-alkilsoderzhashchimi kislotami--allostericheskii protsess.

Mechanism of the activation of 5-lipoxygenase (EC 1.13.11.12) has been investigated and shown to have the allosteric character. Limitations of the activator structure are formulated.

[Hydrophobic nitroxyl radicals inhibit linoleyl alcohol oxidation by 5-lipoxygenase]. / Gidrofobnye nitroksil'nye radikaly--ingibitory okisleniia linolevogo spirta 5-lipoksigenazoi.

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.

[Potato 5-lipoxygenase. Kinetics of linoleic acid oxidation]. / 5-lipoksigenaza kartofelia. Kinetika okisleniia linolevoi kisloty.

The role of main factors influencing the rate of potato 5-lipoxygenase oxidation of linoleic acid was investigated. It was found that nonionic detergent lubrol PX inhibited the potato lipoxygenase. Optimal pH for the linoleic acid oxidation was 6.3 temperature--45 degrees C and substrate concentration--3 x 10(-4) M (if lubrol PX was 0.02%). It was shown that potato 5-lipoxygenase was allosteric enzyme which possessed positive cooperativity for linoleic acid. The Hill coefficient was calculated (n = 1.40 +/- 0.15) with S0.5 = 75 +/- 10 microM.

[biological activity of phosphorus analogs of amino acids]. / Biologicheskaia aktivnost' fosfornykh analogov aminokislot.

Data available in literature on biological activity of the phosphorus analogues of amino acids are summarized. The most typical kinds of bioactivity of aminophosphonic, aminophosphinic and aminophosphonous acids are characterized. These compounds, both natural and synthetic, are shown to possess a wide spectrum of bioactivity and may be estimated as potential bioregulators.

[Activity of lipoxygenase incorporated into reversed micelles of aerosol OT in octane]. / Aktivnost' lipoksigenazy, vkliuchennoi v obrashchennye mitselly aérozolia OT v oktane.

Soybean lipoxygenase (EC 1.13.11.12) incorporated into the reversed micelles of aerosol OT in octane has been studied for its catalytic properties. The enzyme is shown to preserve up to 10% activity as compared with the activity in the aqueous solution. In this case Km of lipoxygenase for linoleic acid increases from 10(-5) M to 5 X 10(-4) M. The activity of lipoxygenase is maximal, the aerosol OT concentration being 0.03 M and a degree of reversed micelle hydratation 40. Cationic detergents of the cetyltrimethyl ammonium bromide type are not good to form reversed micelles of lipoxygenase, since they inhibit the latter with IC50 = (4 divided by 6) x 10(-4) M. The lipoxygenase preparations in reversed micelles of aerosol OT in octane may be used to synthesize natural metabolites of polyunsaturated fatty acids, for instance of eicosanoids.

[Phosphorus-containing inhibitors of penicillin acylase. 3. Inhibition of Escherichia coli penicillin acylase by phosphonate analogs of substrates]. / Fosforsoderzhashchie ingibitory penitsillinatsilazy. 3. Ingibirovanie penitsillinatsilazy Escherichia coli fosfonatnymi analogami substratov.

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.

[Phosphorus-containing inhibitors of penicillin acylase. 2. Design, synthesis and study of the hydrolytic stability of phosphonic acid derivatives as potential inhibitors of penicillin acylase]. / Fosforsoderzhashchie ingibitory penitsillinatsilazy. 2. Dizain, sintez i izuchenie gidroliticheskoi stabil'nosti proizvodnykh fosfonovykh kislot kak potentsial'nykh ingibitorov penitsillinatsilazy.

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.

[Inhibition of penicillin acylase from Escherichia coli by benzylalkylketones and benzylalkylcarbinols]. / Ingibirovanie benzilalkilketonami i benzilalkilkarbinolami penitsillinatsilazy iz Escherichia coli.

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.

[Inhibition of yeast pyruvate decarboxylase by alkyl phosphates]. / Ingibirovanie drozhzhevoi piruvatdekarboksilazy alkilfosfatami.

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.

[Phosphorus-containing inhibitors of penicillin acylase. 4. Irreversible inhibition of penicillin acylase from Escherichia coli by monoaryl esters of benzylphosphonic acid]. / Fosfor-soderzhashchie ingibitory penitsillinatsilazy. 4. Neobratimoe ingibirovanie penitsillinatsilazy iz Escherichia coli monoarilovymi éfirami benzilfosfonovoi kisloty.

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.

[Antithrombic activity of N(alpha)-lauroyl-L-arginine ethyl ester (LAE) and 9-fluorenylmethoxycarbonyl-L-arginine methyl ester (Fmoc-L-Arg-OMe)].

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.

[Calix[4]arene methylenebisphosphonic acids: the features of alkaline phosphatases inhibition].

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.

[Inhibiting properties of stable nitroxyl radicals in reactions of linoleic acid and linoleyl alcohol oxidation catalyzed by 5-lipoxygenase].

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.

The unusual action of (R,S)-2-hydroxy-2-trifluoromethyl-trans-n-octadec-4-enoic acid on 5-lipoxygenase from potato tubers.

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.

O-Perfluoroalkylation of 4-hydroxyphenylglycine.

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.