ABSTRACT
Aliphatic and aromatic mono-, di-, and triesters of phosphonoformic acid (foscarnet) were synthesized. The triesters were prepared by the Michaelis-Arbuzov reaction and were hydrolyzed to di- and monoesters. The compounds were tested for antiviral activity on isolated herpes simplex virus type 1 (HSV-1) DNA polymerase, in a HSV-1 plaque reduction assay, and on a cutaneous HSV-1 infection in guinea pigs. None of the esters inhibited the activity of isolated HSV-1 polymerases. Monoesters with a free carboxylic group and diesters with an aromatic carboxylic ester function were active against the cutaneous herpes infection. Mono- and diesters with an aromatic phosphonic ester group also showed activity in the plaque-reduction assay. However, mono- and diesters with an aromatic phosphonic ester group also showed activity in the plaque-reduction assay. However, mono- and diesters with aliphatic carboxylic ester groups were inactive in all test systems. The results show that all three acidic groups of phosphonoformic acid must be free in order to get antiviral activity at the enzyme level. However, certain esters of this acid may be biotransformed to the acid itself to give antiherpes activity.
Subject(s)
Antiviral Agents/chemical synthesis , Organophosphorus Compounds/chemical synthesis , Phosphonoacetic Acid , Phosphonoacetic Acid/chemical synthesis , Simplexvirus/drug effects , Esters , Foscarnet , Indicators and Reagents , Nucleic Acid Synthesis Inhibitors , Phosphonoacetic Acid/analogs & derivatives , Phosphonoacetic Acid/pharmacology , Simplexvirus/enzymology , Structure-Activity Relationship , Viral Plaque AssayABSTRACT
A DNA polymerase is associated with the core of the so-called Dane particles. The probability that this is the hepatitis B viral DNA polymerase offers the possibility of preventing hepatitis B multiplication by selective inhibition of this enzyme. We have previously reported that trisodium phosphonoformate (PFA) inhibits Dane particle DNA polymerase. Fifteen compounds with structural similarity to PFA and pyrophosphate have now been tested for inhibition of hepatitis B virus DNA polymerase in an attempt to define the structural requirement for the inhibition. Active structures have two acid groups at close proximity of which at least one is a phosphono group. Phosphonoformate and hypophosphare were the two most active inhibitors. The Ki value for PFA was 7.2 microM when dTTP was used as variable substrate, and the mechanism of inhibition was non-competitive. Phosphonoformate caused rapid shut-off of the polymerase reaction, indicating that it might inhibit elongation. The efficient inhibition of hepatitis B virus DNA polymerase by PFA and its low toxicity suggest that it could be used to inhibit hepatitis B virus multiplication in vivo.
Subject(s)
Diphosphates/pharmacology , Hepatitis B virus/enzymology , Nucleic Acid Synthesis Inhibitors , Humans , Structure-Activity Relationship , Time FactorsABSTRACT
Twenty nine pyrophosphate analogues have been tested for their ability to inhibit herpes simplex virus type 1 (HSV-1) DNA polymerase activity. The structural requirements for active inhibitors and their mechanism of action have been investigated. Active compounds had two negatively charged groups at close proximity. The most active compounds contained free phosphono and/or carboxyl groups. Apart from phosphonoformate and phosphonoacetate, which were strong inhibitors, some other analogues, 2-phosphonopropionate, 2-phenylphosphonoacetate, oxalate, carbonyldiphosphate, methanehydroxy-diphosphonate and hypophosphate also inhibited the HSV-1 DNA polymerase activity. With the exception of hypophosphate the tested compounds inhibited the activity of calf thymus DNA polymerase alpha to a lesser extent than the activity of the HSV-1 DNA polymerase. All inhibitors gave linear uncompetitive inhibition of HSV-1 DNA polymerase with activated DNA as variable substrate. With the four deoxynucleoside triphosphates as variable substrates all inhibitors except hypophosphate gave linear non-competitive inhibition patterns. Hypophosphate caused a more competitive type of inhibition. The inhibition constants for the active compounds have been determined. Phosphonoformate, phosphonoacetate, methanehydroxydiphosphonate and hypophosphate also inhibited HSV-1 plaque formation in cell culture.
Subject(s)
Diphosphates/pharmacology , Nucleic Acid Synthesis Inhibitors , Simplexvirus/enzymology , Animals , Cattle , DNA Polymerase II/antagonists & inhibitors , Kinetics , Phosphonoacetic Acid/pharmacology , Structure-Activity RelationshipSubject(s)
Antiviral Agents/pharmacology , Viruses/enzymology , Enzyme Activation/drug effects , Enzyme Induction , Enzyme Inhibitors/pharmacology , Nucleic Acid Synthesis Inhibitors , Oxidoreductases/antagonists & inhibitors , RNA Ligase (ATP)/antagonists & inhibitors , Substrate Specificity , Transferases/antagonists & inhibitorsABSTRACT
Evidence available indicates that the so-called Dane particles are the hepatitis virus. A DNA polymerase is associated with the core of these particles. The probability that this is the viral DNA polymerase offers the possibility of preventing hepatitis B multiplication by selective inhibition of this enzyme. This investigation reports that trisodium phosphonoformate (PFA) at low concentrations but not phosphonoacetate acid (PAA) inhitits Dane particle associated DNA polymerase.
Subject(s)
Formates/pharmacology , Hepatitis B Antigens , Nucleic Acid Synthesis InhibitorsSubject(s)
Herpesviridae/drug effects , Organophosphorus Compounds/pharmacology , Phosphonoacetic Acid/pharmacology , Animals , DNA/biosynthesis , Formates/pharmacology , Formates/therapeutic use , Guinea Pigs , Haplorhini , Herpesviridae Infections/drug therapy , Male , Phosphonoacetic Acid/analogs & derivatives , Phosphonoacetic Acid/therapeutic use , Phosphonoacetic Acid/toxicity , Protein Biosynthesis , RNA/biosynthesis , Virus Replication/drug effectsABSTRACT
Analogues of pyrophosphate have been tested as inhibitors of influenza virus-RNA polymerase activity in cell-free assays. The most active compound, phosphonoformic acid (PFA), reduced the polymerase activity to 50 per cent at a concentration of 20 muM. The inhibition was dependent on the type of divalent cation present in the assay. PFA at a concentration of 400 muM also inhibited the influenza virus plaque formation by 90 per cent.
Subject(s)
DNA-Directed RNA Polymerases/antagonists & inhibitors , Diphosphates/pharmacology , Orthomyxoviridae/drug effects , Magnesium/pharmacology , Manganese/pharmacology , Orthomyxoviridae/enzymology , Orthomyxoviridae/growth & development , Structure-Activity Relationship , Viral Plaque AssayABSTRACT
Trisodium phosphonoformate selectively inhibits cell-free DNA polymerase activity induced by herpesvirus. The new inhibitor has an antiviral effect on herpes simplex virus types 1 and 2, pseudorables virus, and infectious bovine rhinotracheitis virus in cell culture. It has a good therapeutic activity against cutaneous herpes simplex virus infection in guinea pigs.
Subject(s)
Antiviral Agents , DNA-Directed RNA Polymerases/antagonists & inhibitors , Nucleic Acid Synthesis Inhibitors , Organophosphorus Compounds/pharmacology , Animals , Antiviral Agents/therapeutic use , Antiviral Agents/toxicity , Cell Line , Formates/pharmacology , Formates/toxicity , Guinea Pigs , Herpesviridae Infections/drug therapy , Organophosphorus Compounds/toxicity , Phosphonoacetic Acid/pharmacology , Simplexvirus/enzymologyABSTRACT
Ribavirin 5'-triphosphate (RTP), derived from the broad-spectrum antiviral compound ribavirin (Virazole), can selectively inhibit influenza virus ribonucleic acid polymerase in a cell-free assay. Ribavirin and its 5'-monophosphate have no effect on the polymerase. The inhibition is competitive with respect to adenosine 5'-triphosphate and guanosine 5'-triphosphate. RTP also inhibits ApG- and GpC-stimulated influenza virus ribonucleic acid polymerase. Since ribavirin is phosphorylated in the cell, the inhibition of influenza multiplication in the cell may also be caused by RTP.