ABSTRACT
Various methods of synthesis of metabolically stable phosphonate analogues of bisnucleoside oligophosphates containing two residues of methylenediphosphonic acid in the oligophosphate chain are studied. Phosphonate analogues of Ip4I and Ip5I are prepared.
Subject(s)
Inosine Nucleotides/chemical synthesis , Organophosphonates/chemical synthesis , Magnetic Resonance Spectroscopy , Mass Spectrometry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Spectrophotometry, UltravioletABSTRACT
P-(Alkyl)esters of AZT 5'-hydrogenphosphonate were synthesized and their stabilities in the phosphate buffer and human serum were evaluated. The esters bearing residues of primary and secondary alcohols were degraded to give AZT, whereas those containing tertiary alkyl groups yielded AZT 5'-hydrogenphosphonate. The corresponding derivatives of d2A and d4T showed the same properties.
Subject(s)
Anti-HIV Agents/chemistry , Organophosphonates/chemistry , Reverse Transcriptase Inhibitors/chemistry , Zidovudine/administration & dosage , Zidovudine/chemistry , Anti-HIV Agents/administration & dosage , Delayed-Action Preparations , Humans , Magnetic Resonance Spectroscopy , Reverse Transcriptase Inhibitors/administration & dosageABSTRACT
A series of pyrophosphoryl (Z)-(phosphonomethoxy)but-2-enyl derivatives of pyrimidines and purines 9a-d and the corresponding phosphonates 10a-d were synthesized. The prepared compounds contain the phosphonate group as an alpha-phosphate mimic as well as an acyclic residue emulating the sugar moiety in 2',3'-dideoxy-2',3'-didehydronucleoside 5'-triphosphates known as highly potent chain terminators of DNA polymerases. Phosphonates 10a-d were obtained by alternative alkylations of the nucleic bases followed by condensation with ethyl [[(p-tolylsulfonyl)oxy]methyl]phosphonate. Pyrophosphorylation of 10a-d afforded phosphonate diphosphates 9a-d. Their substrate properties were evaluated in cell-free systems containing various DNA polymerases including viral reverse transcriptases. Compounds 9a-d manifested good terminating substrate properties toward HIV-1 and AMV reverse transcriptases. They exhibited high selectivity and were not recognized by human DNA polymerases alpha and epsilon, DNA polymerase beta from rat liver, Escherichia coli DNA polymerase I, and HSV-1 and CMV DNA polymerases. Phosphonates 10b-d displayed no activity in HIV-1-infected MT-4 cells cultures; 10a was moderately effective (ED50 = 9 microM).
Subject(s)
Nucleotides/chemical synthesis , Nucleotides/pharmacology , Animals , Base Sequence , Cells, Cultured , DNA Primers , HIV Reverse Transcriptase , Humans , Molecular Sequence Data , Nucleic Acid Synthesis Inhibitors , Rats , Reverse Transcriptase InhibitorsSubject(s)
Antiviral Agents/chemical synthesis , DNA Replication/drug effects , DNA-Directed DNA Polymerase/metabolism , Deoxyribonucleotides/metabolism , HIV/drug effects , Organophosphonates/metabolism , Animals , Antiviral Agents/metabolism , Antiviral Agents/pharmacology , DNA Nucleotidylexotransferase/metabolism , Deoxyribonucleotides/chemical synthesis , Deoxyribonucleotides/pharmacology , HIV/enzymology , Humans , Indicators and Reagents , Organophosphonates/chemical synthesis , Organophosphonates/pharmacology , RNA-Directed DNA Polymerase/metabolism , Reverse Transcriptase Inhibitors , Substrate SpecificitySubject(s)
Antiviral Agents/pharmacology , HIV/physiology , Lymphocytes/drug effects , Nucleotides/pharmacology , Virus Replication/drug effects , Animals , Antibody Formation/drug effects , Genome, Viral , HIV/drug effects , HIV/genetics , Lymphocyte Activation/drug effects , Lymphocytes/immunology , Mice , Nucleotides/chemical synthesisABSTRACT
The interaction of a number of ATP analogs with a modified triphosphate moiety as well as 2-chloro-ethyl-amino derivatives of nucleotides, 4(N-2-chloroethyl-N-methylamino)-benzyl-gamma-amide of ATP and the corresponding ADP beta-amide with acetyl-CoA carboxylase (EC 6.4.1.2.) from rat liver has been studied. Halophosphonate derivatives of ATP have been synthesized from bromomethylene diphosphonic acid and found to be reversible inhibitors of the enzyme. ATP and ADP alkylating amides have proved to form a reversible complex with the ATP-binding site and to modify a group in the acetyl-CoA-binding site. The bicarbonate ion accelerates the process of inactivation. The estimate of the distance between the ATP-binding site and the acetyl-CoA-binding site ranges within 0.8-1.2 nm.
Subject(s)
Acetyl-CoA Carboxylase/antagonists & inhibitors , Adenosine Triphosphate/analogs & derivatives , Liver/enzymology , Adenosine Triphosphate/metabolism , Affinity Labels , Animals , Bicarbonates/pharmacology , Biotin/metabolism , Kinetics , Models, Molecular , Rats , Substrate SpecificityABSTRACT
The substrate specificity of procaryotic and eucaryotic AppppA-degrading enzymes was investigated with phosphonate analogues of diadenosine 5',5'''-P1,P4-tetraphosphate (AppppA). App(CH2)ppA (I), App(CHBr)ppA (II), and Appp(CH2)pA (III), but not Ap(CH2)pp(CH2)pA (IV), are substrates for lupin AppppA hydrolase (EC 3.6.1.17) and phosphodiesterase I (EC 3.1.4.1). None of the four analogues is hydrolyzed by bacterial AppppA hydrolase (EC 3.6.1.41), and only analogue III is degraded by yeast AppppA phosphorylase (EC 2.7.7.53). The analogues are competitive inhibitors of all four enzymes. The affinity of analogue IV is 3-40-fold lower than that of analogues I-III for all four enzymes. Introduction of one methylene (as in I and III) [or bromomethylene (as in II)] group into AppppA results in a 3-15-fold increase of its affinity for lupin and Escherichia coli AppppA hydrolases. The same modifications only negligibly (10-30%) affect its affinity for yeast AppppA phosphorylase and decrease its affinity for lupin phosphodiesterase I about 2.5-fold. The data provide further evidence for the heterogeneity among catalytic sites of all four AppppA-degrading enzymes.
Subject(s)
Acid Anhydride Hydrolases , Adenine Nucleotides/metabolism , Dinucleoside Phosphates , Nucleotidyltransferases/metabolism , Organophosphonates/metabolism , Phosphoric Diester Hydrolases/metabolism , Phosphoric Monoester Hydrolases/metabolism , Adenine Nucleotides/pharmacology , Escherichia coli/enzymology , Kinetics , Organophosphonates/pharmacology , Phosphodiesterase I , Plants/enzymology , Saccharomyces cerevisiae/enzymology , Structure-Activity Relationship , Substrate SpecificitySubject(s)
Adenosine Monophosphate/analogs & derivatives , Escherichia coli/metabolism , RNA, Transfer, Amino Acyl/metabolism , Ribosomes/metabolism , Adenosine Monophosphate/metabolism , Chemical Phenomena , Chemistry , Chloramphenicol/pharmacology , Lincomycin/pharmacology , Oligopeptides/metabolism , Peptide Biosynthesis , Ribosomes/drug effectsABSTRACT
The model substances 2'(3')-O-[N-acetylmethionylaminomethylene-(P-methyl)phosphino] ester of adenosine-5'-phosphate, pA-(AcMetGlyP), and N-acetylmethionylaminomethylene-(P-methyl) phosphinoamide of phenylalanine, AcMetGly P PheOH, were synthesised. They were used for the ribosomal catalysis studying.
Subject(s)
Adenosine Monophosphate/analogs & derivatives , Escherichia coli/metabolism , Phenylalanine/analogs & derivatives , Ribosomes/metabolism , Drug Evaluation, Preclinical , Ribosomes/drug effectsABSTRACT
The acylamino acid esters of nucleoside 5'-phosphates are synthesized via condensation of N-(N'-acylaminoacyl) imidazoles with nucleoside 5'-phosphates. The PMR and CD spectra of the esters obtained are studied. The 3'-isomers of the substances under study are observed to have a shift in the conformational N in equilibrium S equilibrium of the carbohydrate moiety in favour of the S-form as compared to the initial nucleosides and their 2'-acyl esters.
