WO2003028737A1 - Agents antiviraux et procede in-vitro d'identification de candidats empechant la fixation de la polymerase a l'epsilon - Google Patents

Agents antiviraux et procede in-vitro d'identification de candidats empechant la fixation de la polymerase a l'epsilon Download PDF

Info

Publication number
WO2003028737A1
WO2003028737A1 PCT/JP2002/008799 JP0208799W WO03028737A1 WO 2003028737 A1 WO2003028737 A1 WO 2003028737A1 JP 0208799 W JP0208799 W JP 0208799W WO 03028737 A1 WO03028737 A1 WO 03028737A1
Authority
WO
WIPO (PCT)
Prior art keywords
group
viral
agent according
viral agent
rna
Prior art date
Application number
PCT/JP2002/008799
Other languages
English (en)
Inventor
Satoshi Yuasa
Naohiro Kamiya
Original Assignee
Mitsubishi Pharma Corporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Mitsubishi Pharma Corporation filed Critical Mitsubishi Pharma Corporation
Priority to JP2003532069A priority Critical patent/JP2005508924A/ja
Publication of WO2003028737A1 publication Critical patent/WO2003028737A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A61K31/675—Phosphorus compounds having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A61K31/683—Diesters of a phosphorus acid with two hydroxy compounds, e.g. phosphatidylinositols
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/01—DNA viruses
    • G01N2333/02—Hepadnaviridae, e.g. hepatitis B virus
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/91—Transferases (2.)
    • G01N2333/912—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • G01N2333/91205—Phosphotransferases in general
    • G01N2333/91245—Nucleotidyltransferases (2.7.7)
    • G01N2333/9125—Nucleotidyltransferases (2.7.7) with a definite EC number (2.7.7.-)
    • G01N2333/91255—DNA-directed RNA polymerase (2.7.7.6)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G01N2500/02—Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)

Definitions

  • the present invention relates to an anti-viral agent showing a novel pharmacological action. More specifically, the present invention relates to an anti-viral agent which exerts an inhibitory mechanism of action against activation of viral polymerase via RNA binding. Moreover, the present invention also relates to a method for screening an anti- viral agent by evaluating inhibition of activation of viral polymerase via RNA binding.
  • Hepatitis B virus is a causative agent of an acute or chronic hepatitis B. It is estimated that there are more than 3G0 million HBV carriers throughout the world, and therefore a disease from HBV infection, hepatitis B, is considered to be a serious disease, and has been listed as one of the 10 leading causes of death in the world (World Heath Organization warns of growing "crisis of suffering" http://www.who.int/whr/1997/presse.htm).
  • HBV can infect and replicate only in humans and chimpanzees, a woodchuck hepatitis virus (WHV) and a duck hepatitis B virus (DHBV), which are belonging to Hepadnaviridae like HBV, are recognized as important models of hepadnavirus infection and are used in studies regarding the replication of hepadnaviruses, including searches for the antiviral agents.
  • WBV woodchuck hepatitis virus
  • DHBV duck hepatitis B virus
  • the hepadnavirus uses a pregenome RNA (pgRNA), which encodes its own core protein and polymerase (POL), as an intermediate for replication of its own viral DNA by RNA-dependent DNA polymerase activity of the POL (Cell, Summers and Mason, Vol. 29, 403-415 (1982)).
  • pgRNA pregenome RNA
  • POL polymerase
  • the POL recognizes and binds to a ⁇ motif, a secondary structure of RNA which locates near the 5 '-terminus on a pgRNA, and then viral core particles are formed by encapsidation with core proteins (The EMBO J., Bartenschlager and Schaller, Vol.ll, No.9, 3413-3420 (1992)).
  • DHBV POL is expressed by an in vitro translation system using a rabbit reticulocyte lysate, and then deoxynucleoside triphosphates (dNTPs) are added to a POL- ⁇ RNA complex in the solution to perform priming reaction, followed by detection of POL covalently bonded to labeled dNTP.
  • dNTPs deoxynucleoside triphosphates
  • An object of the present invention is to search for and identify a medicament which inhibits activation of viral polymerase via RNA binding. Another object of the present invention is to provide an anti-viral agent using a medicament which inhibits activation of viral polymerase via RNA binding. A further object of the present invention is to provide a method for screening a medicament which inhibits activation of viral polymerase via RNA binding.
  • a test medicament is added after formation of a POL- ⁇ RNA complex to analyze competitive action with dNTP.
  • the present inventors have improved the conventional experiment system by providing separately each of POL, ⁇ RNA and adding a test medicament before formation of a POL- ⁇ RNA complex, and thereby succeeded to find an inhibitory activity of a compound which did not show the inhibitory activity in the conventional experiment system, The present inventors also succeeded in identifying a novel inhibitory mechanism of viral polymerase, thereby completing the present invention.
  • an anti-viral agent comprising, as an active ingredient, a medicament which inhibits activation of viral polymerase via RNA binding.
  • activation of viral polymerase via RNA binding is activation of viral polymerase via binding of virus RNA and viral polymerase.
  • RNA is ⁇ RNA
  • a pharmacological efficacy is sustained after withdrawing the agent.
  • the virus belongs to Hepadnaviridae, and particularly preferably the virus is a hepatitis B virus.
  • the anti-viral agent wherein the medicament is a phosphonate nucleotide compound represented by the following formula (I) or a salt thereof, or a hydrate or solvate thereof:
  • R is a hydroxyl group or a C ⁇ -C 6 alkoxy group; each of R and R is independently a hydrogen atom, a -C 22 alkyl group, an acyloxymethyl group, an acylthioethyl group, or an ethyl group substituted by one or more halogen atoms; R 4 is a hydrogen atom, a C 1 -C 4 alkyl group, a C ⁇ -C 4 hydroxyalkyl group, or a C ⁇ -C 4 alkyl group substituted by one or more halogen atoms; and X is CH or a nitrogen atom.
  • R 1 is a hydroxyl group or a methoxy group.
  • each of R 2 and R 3 is independently a hydrogen atom, a C1-C 22 alkyl group, or an ethyl group substituted by one or more halogen atoms.
  • each of R 2 and R 3 is independently a hydrogen atom or a 2,2,2-trifluoroethyl group.
  • R 4 is a hydrogen atom or a methyl group.
  • X is CH.
  • R 1 is a hydroxyl group or a methoxy group
  • each of R 2 and R is independently a hydrogen atom or a 2,2,2-trifluoroethyl group
  • R is a hydrogen atom
  • X is CH.
  • the medicament is
  • a method for screening an anti- viral agent which comprises a step of evaluating activation of viral polymerase via RNA binding.
  • the step of evaluating activation of viral polymerase via RNA binding is that of evaluating the degree of binding of virus RNA and viral polymerase.
  • the method for screening an anti-viral agent comprises steps of adding a test medicament to viral polymerase, adding virus RNA, and evaluating the degree of binding of virus RNA and viral polymerase.
  • the method for screening an anti-viral agent comprises steps of adding a test medicament to viral polymerase, adding virus RNA, and judging that the test medicament is an anti- viral agent when the test medicament inhibits the binding of virus RNA and viral polymerase.
  • RNA is ⁇ RNA.
  • the virus belongs to Hepadnaviridae.
  • the virus is a hepatitis B virus.
  • an anti- viral agent obtained by the method for screening an anti- viral agent according to the present invention.
  • an anti-viral agent obtained by the steps of conducting the method for screening an anti-viral agent according to the present invention to obtain an anti-viral substance, producing the thus obtained anti-viral substance by chemical synthesis, and mixing the anti- viral substance with a pharmaceutically acceptable carrier.
  • a method for inhibiting a virus which comprises administration of a pharmacologically effective amount of medicament inhibiting a process where viral polymerase is activated via binding of virus RNA and viral polymerase, to mammals including a human.
  • a medicament inhibiting a process where viral polymerase is activated via binding of virus RNA and viral polymerase, for the production of an anti- viral agent.
  • Figure 1 shows a schematic diagram of a plasmid construct.
  • A shows DHBV POL3' ⁇ used in the conventional method
  • B shows DHBV POL ⁇ (-) used in the improved method for the preparation of POL
  • C shows DHBV 5' ⁇ " used in the improved method as ⁇ RNA
  • D shows DHBV HIS-TP used in the improved method for the preparation of Terminal Protein (TP)
  • E shows DHBV ⁇ TP-POL ⁇ (-) used in the improved method for the preparation of TP-deleted POL.
  • FIG. 2 shows test results obtained by the conventional method.
  • Figure 3 shows test results in respect of RNA concentration dependency by an improved method.
  • Figure 4 shows results of ⁇ RNA competition tests obtained by an improved method.
  • Figure 5 shows test results in respect of the analysis of POL level dependency by an improved method.
  • Figure 6 shows results of study of the TP or ⁇ RNA concentration dependency.
  • Figure 7 shows results of inhibitory activity of medicament against TP as protein primer during the formation of TP- ⁇ TP POL- ⁇ RNA complex.
  • the anti-viral agent of the present invention is characterized in that it comprises, as an active ingredient, a medicament which inhibits a process where viral polymerase is activated via binding of virus RNA and viral polymerase, and preferably that the pharmacological efficacy is sustained after withdrawing the agent.
  • medicaments include the phosphonate nucleotide compound of the above formula (I) or a salt thereof, or a hydrate or solvate thereof, and the lik .
  • examples of a C ⁇ -C 6 alkoxy group represented by R 1 include a methoxy group, an ethoxy group, an n-propyloxy group, an isopropyloxy group, an n-butyloxy group, an isobutyloxy group, a sec-butyloxy group, a tert-butyloxy group, a pentyloxy group, a hexyloxy group, and the like.
  • examples of a C 1 -C 22 alkyl group represented by R 2 and R 3 include a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, a pentyl group, a hexyl group, a heptyl group, an octyl group, a nonyl group, a decyl group, an undecyl group, a dodecyl group, a tridecyl group, a tetradecyl group, a pentadecyl group, a hexadecyl group, a heptadecyl group, an octadecyl group, a nonadecyl group, an i
  • Examples of an acyloxymethyl group represented by R 2 and R 3 include an acetyloxymethyl group, a propionyloxymethyl group, a butyryloxymethyl group, an isobutyryloxymethyl group, a valeryloxymethyl group, an isovaleryloxymethyl group, a pivaloyloxymethyl group, and the like.
  • Examples of an acylthioethyl group represented by R 2 and R 3 include an acetylthioethyl group, a propionylthioethyl group, a butyrylthioethyl group, an isobutyrylthioethyl group, a valerylthioethyl group, an isovalerylthioethyl group, a pivaloylthioethyl group, and the like.
  • the type of the halogen atom may be any of a fluorine, chlorine, bromine or iodine atom.
  • Examples of an ethyl group substituted by one or more halogen atoms include a 1-fluoroethyl group, a 2-fluoroethyl group, a 1-chloroethyl group, a 2-chloroethyl group, a 2-bromoethyl group, a 2,2-difluoroethyl group, a 2,2-dichloroethyl group, a 2,2-dibromoethyl group, a 2,2,2-trifluoroethyl group, a 2,2,2-trichloroethyl group, a 2,2,2-tribromoethyl group, and the like. It is particularly preferred that the 2-position of an ethyl group is substituted
  • R 9 atom is a fluorine atom.
  • At least one of R and R is preferably an ethyl group substituted by one or more halogen atoms, and particularly preferably 2,2,2-trifluoroethyl group.
  • Examples of a C ⁇ -C alkyl group represented by R include a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, and the like.
  • Ci-C 4 hydroxyalkyl group represented by R 4 examples include a hydroxymethyl group, a 1-hydroxyethyl group, a 2-hydroxyethyl group, a 1-hydroxypropyl group, a 2-hydroxypro ⁇ yl group, a 3-hydroxypropyl group, a 1-hydroxybutyl group, a 2-hydroxybutyl group, a 3-hydroxybutyl group, a 4-hydroxybutyl group, and the like.
  • Examples of a - alkyl group substituted by one or more halogen atoms represented by R 4 include a group in which a halogen atom(s) such as a fluorine atom or a chlorine atom is/are bound to a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, and the like.
  • a halogen atom(s) such as a fluorine atom or a chlorine atom
  • Such a group include a fluoromethyl group, a difluoromethyl group, a trifluoromethyl group, a fluoroethyl group, a chloroethyl group, a fluoropropyl group, a chloropropyl group, a fluorobutyl group, a chlorobutyl group, and the like.
  • the first condition for the preferred compound of the present invention is that each of R 2 and R 3 is independently a hydrogen atom, a -C2 alkyl group or an ethyl group substituted by one or more halogen atoms.
  • the second condition for the preferred compound of the present invention is that R 2 and R 3 is independently a hydrogen atom, a -C 22 alkyl group or a 2,2,2-trifluoroethyl group, and R 4 is a hydrogen atom or a methyl group.
  • Examples of specific preferred compounds satisfying such conditions include the following compounds:
  • each of R 2 and R 3 is a 2,2,2-trifluoroethyl group and R 4 is a hydrogen atom or a methyl group.
  • preferred compounds satisfying such conditions include the following compounds: 2-arruno-6-(4-memoxyphenylthio)-9-[2-(phosphonome oxy)ethyl]purine bis(2,2,2- trifluoroethyl) ester;
  • the fourth condition for the preferred compound of the present invention is that R 1 is a hydroxyl group, each of R 2 and R 3 is a 2,2,2-trifluoroethyl group, and R 4 is a hydrogen atom.
  • R 1 is a hydroxyl group
  • R 2 and R 3 is a 2,2,2-trifluoroethyl group
  • R 4 is a hydrogen atom. Examples of preferred compounds satisfying such conditions include the following compounds:
  • the phosphonate nucleotide compound of the above formula (I) of the present invention may exist as a salt, and any salt formed by the above compound can be used as an active ingredient of the anti-viral agent of the present invention.
  • a salt include a pharmaceutically acceptable salt.
  • the acidic group is able to form metal salts such as a lithium salt, a sodium salt, a potassium salt, a magnesium salt and a calcium salt, and ammonium salts such as an ammonium salt, a methyl ammonium salt, a dimethyl ammonium salt, a trimethyl ammonium salt and dicyclohexyl ammonium salt.
  • the arnino group is able to form mineral acid salts such as hydrochloride, hydrobromide, sulfate, nitrate, phosphate and metaphosphate, and organic acid salts such as methanesulfonate, benzenesulfonate, para-toluenesulfonate, acetate, propionate, tartrate, fumarate, maleate, malate, oxalate, succinate, citrate, benzoate, mandelate, cinnamate, lactate, besylate, valerate, stearate, oleate, lactobionate, ethylsuccinate, semisuccinate, butyrate, palmitate, carbamate, gluconate, laurate, salicylate, laokurate, tannate and butylsulfonate.
  • mineral acid salts such as hydrochloride, hydrobromide, sulfate, nitrate, phosphate and metaphosphate
  • the phosphonate nucleotide compound of the above formula (I) and salt thereof may exist in the form of a hydrate or solvate. Any given hydrate or solvate which is formed by the phosphonate nucleotide compound of the above formula (I) or salt thereof, can be used as an active ingredient of the medicament of the present invention.
  • a solvent capable of forming the solvate include methanol, ethanol, isopropanol, acetone, ethyl acetate, methylene chloride, diisopropyl ether, and the like.
  • each of R 2 and R 3 is a C 1 -C 22 alkyl group, or an ethyl group substituted by one or more halogen atoms
  • the compound can be synthesized, for example, according to the following reaction route (1) or (2).
  • R 1 , R 4 and X are the same as defined above, R 5 represents a -C 22 alkyl group, or an ethyl group substimted by one or more halogen atoms, and W represents a leaving group such as a halogen atom, a para-toluenesulfonyloxy group, a methanesulfonyloxy group or a trifluoromethanesulfonyloxy group.
  • the compound of the above formula (II) is reacted with the compound of the above formula (ffl) at a temperature of 10°C to 250°C, preferably 130°C to 200°C, for 0.1 to 100 hours, preferably for 3 to 24 hours.
  • the compound of the above formula (TV) obtained by the above reaction can be separated and purified by ordinary separation and purification means such as distillation-, adsorption- or partition-chromatographies, as necessary.
  • the compound of the above formula (IV) may be separated and purified as stated above, or it may directly be used for the following reaction without purification.
  • the compound of the above formula (IV) is reacted with the compound of the above formula (V) in the presence of a base such as sodium carbonate, potassium carbonate, cesium carbonate, sodium hydrate, potassium hydrate, trie ylamine and diazabicycloundecen in a suitable solvent such as acetonitrile, tetrahydrofuran, dimethylsulfoxide, dimethylformamide or methylpyrrolidone at a temperature of 10°C to 200°C, preferably 50°C to 150°C, for 0.1 to 100 hours, preferably for 1 to 10 hours, to obtain the compound of the above formula (F).
  • a base such as sodium carbonate, potassium carbonate, cesium carbonate, sodium hydrate, potassium hydrate, trie ylamine and diazabicycloundecen
  • a suitable solvent such as acetonitrile, tetrahydrofuran, dimethylsulfoxide, dimethylformamide or methylpyrrolidone
  • the source of the compounds of the above formulas (II), (HI) and (V) which are raw materials for reaction route (1) is not particularly limited.
  • a compound commercially available as a reagent may be used, or a compound may be synthesized by a known method, as appropriate.
  • the compound of the above formula (V) can be synthesized by heating the compound of the formula (VI) and the compound of the formula (VET) which are described later, in a suitable solvent such as acetonitrile or dimethylsulfoxide at a range of 50°C to 100°C.
  • the compound of the above formula (I') can also be produced by the following method.
  • R 1 , R 4 , R 5 , X and W are the same as defined above.
  • the compound of the above formula (IV) obtained by reaction route (1) is reacted with the compound of the above formula (VI) in the presence of a base such as sodium carbonate, potassium carbonate, cesium carbonate, sodium hydrate, potassium hydrate, triemylamine and diazabicycloundecen in a suitable solvent such as acetonitrile, tetrahydrofuran, dimethylsulfoxide, dimethylformamide or methylpyrrolidone at a temperature of 10°C to 200°C, preferably 50°C to 150°C for 0.1 to 100 hours, preferably for 0.5 to 10 hours, to obtain the compound of the above formula (VII).
  • a base such as sodium carbonate, potassium carbonate, cesium carbonate, sodium hydrate, potassium hydrate, triemylamine and diazabicycloundecen
  • a suitable solvent such as acetonitrile, tetrahydrofuran, dimethylsulfoxide, dimethylformamide or methylpyrrolidone
  • the compound of the above formula (NH) is reacted with a mercaptan represented by the above formula (NHJ) or a salt thereof such as a sodium salt, a potassium salt, a lithium salt or a triemylamine salt, in a suitable solvent such as acetonitrile, tetrahydrofuran, dimethylsulfoxide, dimemylformamide or methylpyrrolidone optionally in the presence of a suitable tertiary amine at a temperature of 10°C to 200°C, preferably 70°C to 120°C for 0.1 to 100 hours, preferably for 0.5 to 12 hours, to obtain the compound of the above formula (F).
  • a suitable solvent such as acetonitrile, tetrahydrofuran, dimethylsulfoxide, dimemylformamide or methylpyrrolidone
  • a suitable tertiary amine at a temperature of 10°C to 200°C, preferably 70°C to 120°C for 0.1 to 100
  • the compound of the formula (F) corresponds to a compound of the formula (I) wherein each of R 2 and R 3 is a C 1 -C 22 alkyl group, or an ethyl group substituted by one or more halogen atoms.
  • the source of the compound of the above formula (NT) which is a raw material of reaction route (2) is not particularly limited.
  • a compound commercially available as a reagent may be used, or the compound may be synthesized by a known method as appropriate.
  • a compound of the formula (I) wherein R 3 is a hydrogen atom, a -C 22 alkyl group, an acylthioethyl group, or an ethyl group substituted by one or more halogen atoms, and R 2 is a C 1 -C 22 alkyl group, or an ethyl group substituted by one or more halogen atoms, can be obtained by reaction of the compound of the above formula (F) with the compound of the formula (IX): R 6 OH wherein R 6 is a hydrogen atom, a -C 2 2 alkyl group, an acylthioethyl group, or an ethyl group substituted by one or more halogen atoms, in no solvent or in a suitable solvent including a chloric solvent such as dichloromethane; pyridine; acetonitrile; tetrahydrofuran; dimethylsulfoxide; dimethylformamide and methylpyrrolidone, optional
  • R 1 , R 4 , R 5 , R 6 and X are the same as defined above.
  • a compound of the formula (I) wherein each of R 2 and R 3 is independently a hydrogen atom, a -C 22 alkyl group, an acylthioethyl group, or an ethyl group substituted by one or more halogen atoms can also be obtained by the following method.
  • R 1 , R 4 and X are the same as defined above, and each of R 7 and R 8 is independently a hydrogen atom, a -C 22 alkyl group, an acylthioethyl group, or an ethyl group substituted by one or more halogen atoms, with the exception that both R 7 and R 8 can not represent hydrogen atoms at the same time.
  • the compound of the above formula (I") is reacted with trime ylsilyldiemyla ⁇ iine in a suitable solvent such as dichloromethane, dichloroethane and chloroform around room temperature for about 1 hour. More than two moles of trime ylsUyldiemylamine are used per mole of the compound of the above formula (I").
  • reaction solution is concentrated to dryness, the residue is dissolved into a suitable solvent, for example, a chloric solvent such as dichloromethane, and then oxalyl chloride is added in an amount of 2 or more moles per mole of the compound of the above formula (I"), followed by reaction on ice for about 1 hour, and then around room temperature for about 1 hour in the presence of a catalytic amount of dimethylformamide.
  • a suitable solvent for example, a chloric solvent such as dichloromethane
  • the compound of the above formula (X) obtained by removal of the solvent, usually without being purified, is reacted with the compound of the formula (XI) and/or the compound of the formula (XII) in a suitable solvent, for example, a chloric solvent such as dichloromethane, pyridine, acetonitrile, tetrahydrofuran, dimethylsulfoxide, dimethylformamide or methylpyrrolidone, at a temperature of 10°C to 100°C, preferably 20°C to 30°C for 0.1 to 100 hours, preferably 5 to 12 hours.
  • a chloric solvent such as dichloromethane, pyridine, acetonitrile, tetrahydrofuran, dimethylsulfoxide, dimethylformamide or methylpyrrolidone
  • the obtained compound of the formula (XIII) corresponds to a compound of the formula (I) wherein each of R 2 and R 3 is independently a hydrogen atom, a C1-C 22 alkyl group, an acylthioethyl group, or an ethyl group substituted by one or more halogen atoms with the exception that both R 2 and R 3 can not represent hydrogen atoms at the same time.
  • the compound of the above formula (I") which is a raw material of the above reaction, can be obtained by hydrolysis of the compound of the formula (F), or it can be more efficiently obtained by reaction of a compound of the formula (F) wherein R 5 is a -C 22 alkyl group with triethyliodosilane, trimethylbromosilane and the like.
  • a compound of the formula (I) wherein each of R 2 and R 3 is an acyloxymethyl group, or wherein either one of R 2 and R 3 is an acyloxymethyl group and the other is hydrogen, can be obtained by reaction of the compound of the above formula (I") with an acyloxymethyl halide represented by the following formula (XrV): R 9 Y wherein R 9 is an acyloxymethyl group and Y is a chlorine, bromine or iodine atom, in the presence of a base such as sodium carbonate, potassium carbonate, cesium carbonate, sodium hydride, potassium hydride, triethylamine, pyridine, diazabicycloundecen and N,N'-dichlorohexyl-4-morpholinecarboxamidine in a suitable solvent such as acetonitrile, tetrahydrofuran, dimethylsulfoxide, dimethylformamide or methylpyrrolidone at a temperature of 0°C to 200°C,
  • the compound of the formula (XrV) may be reacted with the compound of the formula (I") in an amount of 2 moles per mole of the compound of the formula (I"), while in the case of a compound wherein either one is an acyloxymethyl group, an equivalent mole reaction may be applied.
  • a compound wherein either one of R 2 and R 3 is an acyloxymethyl group, and the other is a -C22 alkyl group, an acylthioethyl group, or an ethyl group substituted by one or more halogen atoms can be produced by preparing a compound wherein either one of R 2 and R 3 is a C1-C 22 alkyl group, an acylthioethyl, group or an ethyl group substituted by one or more halogen atoms, and the other is a hydrogen atom, and then reacting the compound of the above formula (XIV) to this compound by the above method.
  • the salt of the compound of the formula (I) can be synthesized, for example, by the following method.
  • the compound of the formula (F) is reacted with a corresponding acid with stirring at a temperature of -10°C to 100°C, preferably 10°C to 50°C for 0.1 to 20 hours, preferably for 0.3 to 1 hour in a suitable solvent such as ethyl acetate, isopropanol, acetonitrile, tetrahydiOfuran, dimethylsulfoxide, dimethylformamide or methylpyrrolidone.
  • a suitable solvent such as ethyl acetate, isopropanol, acetonitrile, tetrahydiOfuran, dimethylsulfoxide, dimethylformamide or methylpyrrolidone.
  • the above production method is provided as an example of a method for producing the compound of general formula (I), and so the method for producing the compound used in the present invention is not limited thereto.
  • the compound of the above formula (I) produced by the above method or a salt thereof can be separated and purified by ordinary nucleotide separation and purification means, e.g., by selecting and applying means such as recrystallization-, adsorption-, ion exchange- and partition- chromatographies, as appropriate.
  • a target virus to which the anti-viral agent of the present invention is applicable is not particularly hmited, and specific examples of the target virus include an RNA virus such as human immunodeficiency virus, influenza virus and hepatitis C virus; and a DNA virus such as herpes simplex virus type 1 and type 2, cytomegalovirus, varicella-zoster virus and hepatitis B virus, with hepatitis B virus being more preferable.
  • RNA virus such as human immunodeficiency virus, influenza virus and hepatitis C virus
  • a DNA virus such as herpes simplex virus type 1 and type 2, cytomegalovirus, varicella-zoster virus and hepatitis B virus, with hepatitis B virus being more preferable.
  • the compound of the formula (I) may be used singly, but it is preferred that, using a pharmacologically acceptable pharmaceutical additive, a pharmaceutical composition comprising the above compound as an active ingredient is produced and administered.
  • the composition of the pharmaceutical composition is determined by the solubility of the compound, chemical properties, administration route, dosage regimen and the like.
  • the compound can be orally administered in an dosage form of a granule, a parvule, a powder, a tablet, a hard syrup, a soft capsule, a troche, a syrup, an emulsion, a soft gelatine capsule, a gel, a paste, a suspension, a liposome and the like, or the compound can be administered intravenously, intramuscularly or subcutaneously in the form of an injection.
  • the compound may be formulated into powders for injection, and a solution may be prepared before use.
  • an organic or inorganic, solid or liquid carrier which is suitable for oral, enteral, parenteral or local administration
  • a solid carrier used for the production of a solid formulation include lactose, sucrose, starch, talc, cellulose, dextrin, kaoline, calcium carbonate, agar, pectin, stearic acid, magnesium stearate, lecithin, and sodium chloride.
  • a liquid carrier used for the production of a liquid formulation for oral administration include glycerine, peanut oil, polyvinylpyrrolidone, olive oil, ethanol, benzyl alcohol, propylene glycol, physiological saline, and water.
  • the above pharmaceutical composition can also comprise, in addition to the above carriers, an adjuvant such as a wetting agent, a suspension aid, a sweetener, a flavor, a coloring agent and a preservative.
  • a liquid agent may be contained in a capsule of a substance which can be absorbed, such as gelatin.
  • a solvent or a suspending agent which is used for the production of a formulation for parenteral administration such as an injection, include water, propylene glycol, polyethylene glycol, benzyl alcohol, ethyl oleate, and lecithin.
  • the compound of the formula (T), especially the ester derivative of the above formula (F) has a high oral absorbency, and therefore oral administration is a preferred administration route for the anti- viral agent of the present invention.
  • the preparation of each of the above formulation can be carried out according to standard techniques.
  • the anti- viral agent of the present invention is used for oral administration, the clinical dose is generally 0.1 to 500mg of the compound per kg adult per day, and preferably 0.1 to 50mg of the compound per kg adult per day. The dose may be changed as appropriate, depending on age, disease condition, symptom, the presence or absence of concurrent administration and the like.
  • the above dose may be applied once a day or divided over two to several administrations per day at regular intervals, or may also be applied intermittently every several days.
  • the applied dose is 0.01 to 50mg of the compound per kg adult per day, preferably 0.1 to 5mg per kg.
  • the present invention is further described in the following examples.
  • the present invention is not limited to the Examples.
  • the compound numbers in the Examples correspond to those in Table 1.
  • 2-(phosphonomethoxy)ethyliodo bis(2,2,2-trifluoroethyl) ester was added to the above reaction solution, and the mixture was reacted at 100°C for 5 hours. After reaction, the mixture was cooled to room temperature, followed by concentration to dryness. The residue was dissolved in chloroform, and was then adsorbed to a silica gel column followed by elution with 5%-methanol-chloroform to obtain 23.3g (yield 56%) of 2-an ⁇ o-6-chloro-9-[2-(phosphonomemoxy)emyl]purme bis(2,2,2-trifluoroethyl) ester.
  • 2-amino-6-chloro-9- [2-(phosphonomethoxy)ethyl]purine bisisopropyl ester was obtained by the same process as in Example 1, with the only exception being that triisopropylphosphate was used for substitution of Tris(2,2,2-trifluoroethyl)phosphate.
  • FIG. 1 The schematic diagram of plasmid constructs used in this test is shown in Figure 1.
  • Each of the plasmid constructs has a T7, T3 or Sp6 promoter region upstream of a region encoding an RNA of interest.
  • an RNA of interest was obtained.
  • both a region encoding POL and a ⁇ motif exist in the same RNA.
  • an RNA encoding POL and another RNA having a ⁇ motif were synthesized separately. Free nucleotides were removed from the obtained RNA products by MicroSpin S-300HR column (Amersham), and the concentration of RNA was adjusted with nuclease-free water.
  • the oral prodrug of phosphonomethoxyethyl adenine (PMEA; general name: adefovir) is undergoing a clinical test for an anti-HBN agent; and the diphosphorylated form of PMEA (PMEApp) competes with dATP and proved to be an active metabolite of PMEA.
  • PMEApp diphosphorylated form of PMEA
  • compound A (2-amino-6-(4-hydroxyphenylthio) 9-[2-(phosphonomethoxy)ethyl]purine) (described in Japanese Patent Application Laid-Open (Kokai) No. 9-255695 (Japanese Patent No. 3148139)
  • ⁇ RNA obtained from the construct shown in Figure 1C was added to the solution at final concentration of 500 to 5.12nM and the mixture was further incubated at 30°C for 60 minutes.
  • 50mM Tris-hydrochloride buffer (pH7.5) 15mM NaCl and lOmM MgCl 2 at final concentration were added, and [ a 32P]-dGTP (600Ci/mmol) was further added thereto at final concentration of 1.6 ⁇ M to prepare the mixture of a final volume of 5 ⁇ 1, followed by incubation at 30°C for 30 minutes.
  • test by the improved method was carried out by preparing POL according to the above-described method, adding cycloheximide thereto at final concentration of lmM, and further adding a test medicament, followed by incubation at 30°C for 15 to 30 minutes.
  • the test medicament used in the present invention were compound A (2-ammo-6-(4-memoxyphenylmio)-9-[2-(phosphonomethoxy)ethyl]purine) (described in Japanese Patent Application Laid-Open (Kokai) No. 9-255695 (Japanese Patent No.
  • RNA was added thereto and the mixture was further incubated at 30°C for 60 minutes.
  • 50mM Tris-hydrochloride buffer (pH7.5), 15mM NaCl and lOmM MgCl 2 at final concentration were added to 2.5 ⁇ 1 of the obtained reaction solution, and dNTP was further added thereto to obtain the mixture of a final volume of 5 ⁇ 1, followed by incubation at 30°C for 30 minutes.
  • either [ 32P] labeled dATP or dGTP with a specific activity of 400 to l,000Ci/mmol was prepared and used.
  • Terminal Protein (TP) region to which deoxynucleotide is bound is bound
  • Reversetranscriptase (RT) region having a reversetranscriptase activity is present on POL of Hepadnaviridae virus.
  • TP region and a region other than TP on POL were translated from the separate RNAs by using the constructs shown in Figure 1 D and E, and the formation of TP- ⁇ TP POL- ⁇ RNA complex was investigated.
  • RNA derived from the construct of Figure IE was fixed to be l.O g/ l and the final concentration of RNA derived from the construct of Figure ID is adjusted to be 1.0 to 0.2 ⁇ g/ ⁇ 1.
  • ⁇ RNA obtained from the construct of Figure 1C was added to the obtained solution at a final concentration of 600 to 20 nM in 1/10 volume, and the mixture was incubated at 30°C for 60 minutes.
  • Inhibitory activity of medicament against the formation of TP- ⁇ TP POL- ⁇ RNA complex was assayed by performing the aforementioned translation reaction in the presence of a test medicament to bind ⁇ RNA at a concentration of lOOnM and detecting deoxynucleotide binding reaction and TP.
  • Compound A, Compound B, PMEA and PMEG (9-(2-phosphonomemoxyethyl)guanine) were used as test medicaments.
  • Compound A, Compound B and PMEG which are 2-a ⁇ inopurine derivatives inhibited the binding of deoxynucleotide to TP, but PMEA showed no strong inhibition under the same condition.
  • the structure of Compound C is shown below.
  • a serum WHV DNA level was determined by slot blot method. After the WHV DNA level (pg/ml) was normalized by logarithmic transformation, p value was obtained by Dunnett's test between the mean value in each test group and a placebo control group at various points. The results are shown in the following table 3. After the administration of the medicament was terminated after 28 days, in respect of a lOmg/body weight kg/day administration group, significant decrease of the WHV DNA level was observed until the 112 th day, the termination of the observation.
  • N 3 lOmg of Compound C was administered from the 84 th day
  • compounds A, B and C have a profile different from PMEA and its active body, PMEApp, and therefore these compounds are polymerase inhibitors having a novel action mechanism which is different from a competitive action with dNTP. It is considered that the compounds act at the initial stage of the replication of a hepadnaviruses, and inhibit the activation of polymerase by formation of a POL- ⁇ RNA complex
  • the existing anti-viral agent which is competitive with dNTP has a problem regarding rebound of virus level after drug withdrawal, while the medicament of the present invention, which appears to have a polymerase inactivation action, is expected to have a sustained pharmaceutical effect even after drug withdrawal.
  • the anti- viral agent of the present invention has an excellent anti- viral activity, a high oral absorbency and high safety to organisms. Moreover, the anti-viral agent of the present invention which has a polymerase inactivation action, has a sustained pharmaceutical effect even after drug withdrawal, that is, after the termination of administration of the medicament.

Abstract

L'invention a pour but la détection et l'identification d'un médicament empêchant l'activation de la polymérase virale par fixation de l'ARN, et également la formation d'un agent antiviral empêchant l'activation de la polymérase virale par fixation de l'ARN. L'invention porte également sur un agent antiviral comportant comme principe actif un médicament empêchant l'activation de la polymérise virale par fixation de l'ARN.
PCT/JP2002/008799 2001-08-30 2002-08-30 Agents antiviraux et procede in-vitro d'identification de candidats empechant la fixation de la polymerase a l'epsilon WO2003028737A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2003532069A JP2005508924A (ja) 2001-08-30 2002-08-30 抗ウイルス剤

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2001262437 2001-08-30
JP2001-262437 2001-08-30

Publications (1)

Publication Number Publication Date
WO2003028737A1 true WO2003028737A1 (fr) 2003-04-10

Family

ID=19089337

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2002/008799 WO2003028737A1 (fr) 2001-08-30 2002-08-30 Agents antiviraux et procede in-vitro d'identification de candidats empechant la fixation de la polymerase a l'epsilon

Country Status (2)

Country Link
JP (1) JP2005508924A (fr)
WO (1) WO2003028737A1 (fr)

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004046157A1 (fr) * 2002-11-15 2004-06-03 Mitsubishi Pharma Corporation Procede servant a preparer halogenure de 2-[bis(2,2,2-trifluoroethyl)phosphonylmethoxy]ethyl
WO2005002626A2 (fr) * 2003-04-25 2005-01-13 Gilead Sciences, Inc. Composes de phosphonate therapeutiques
WO2004096286A3 (fr) * 2003-04-25 2005-06-16 Gilead Sciences Inc Analogues de phosphonate antiviraux
US7273716B2 (en) 2003-04-25 2007-09-25 Gilead Sciences, Inc. Methods and compositions for identifying therapeutic compounds with GS-7340 ester hydrolase
US7300924B2 (en) 2003-04-25 2007-11-27 Gilead Sciences, Inc. Anti-infective phosphonate analogs
US7407965B2 (en) 2003-04-25 2008-08-05 Gilead Sciences, Inc. Phosphonate analogs for treating metabolic diseases
US7417055B2 (en) 2003-04-25 2008-08-26 Gilead Sciences, Inc. Kinase inhibitory phosphonate analogs
US7427624B2 (en) 2003-10-24 2008-09-23 Gilead Sciences, Inc. Purine nucleoside phosphorylase inhibitory phosphonate compounds
US7427636B2 (en) 2003-04-25 2008-09-23 Gilead Sciences, Inc. Inosine monophosphate dehydrogenase inhibitory phosphonate compounds
US7432273B2 (en) 2003-10-24 2008-10-07 Gilead Sciences, Inc. Phosphonate analogs of antimetabolites
US7432261B2 (en) 2003-04-25 2008-10-07 Gilead Sciences, Inc. Anti-inflammatory phosphonate compounds
US7432272B2 (en) 2003-12-22 2008-10-07 Gilead Sciences, Inc. Antiviral analogs
US7452901B2 (en) 2003-04-25 2008-11-18 Gilead Sciences, Inc. Anti-cancer phosphonate analogs
US7462608B2 (en) 2002-04-26 2008-12-09 Gilead Sciences, Inc. Non nucleoside reverse transcriptase inhibitors
US7470724B2 (en) 2003-04-25 2008-12-30 Gilead Sciences, Inc. Phosphonate compounds having immuno-modulatory activity
KR101154532B1 (ko) 2003-11-12 2012-07-04 길리애드 사이언시즈, 인코포레이티드 항바이러스 포스포네이트 유사체
US8951986B2 (en) 2008-07-08 2015-02-10 Gilead Sciences, Inc. Salts of HIV inhibitor compounds
US9187507B2 (en) 2009-12-10 2015-11-17 Institute Of Pharmacology And Toxicology Academy Of Military Medical Sciences P.L.A. China Acyclic nucleoside phosphonate derivatives and medical uses thereof
US9457035B2 (en) 2004-07-27 2016-10-04 Gilead Sciences, Inc. Antiviral compounds
US9593137B2 (en) 2011-12-22 2017-03-14 Geron Corporation Guanine analogs as telomerase substrates and telomere length affectors
US10851125B2 (en) 2017-08-01 2020-12-01 Gilead Sciences, Inc. Crystalline forms of ethyl ((S)-((((2R,5R)-5-(6-amino-9H-purin-9-yl)-4-fluoro-2,5-dihydrofuran-2-yl)oxy)methyl)(phenoxy)phosphoryl(-L-alaninate

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994003467A2 (fr) * 1992-08-05 1994-02-17 Institute Of Organic Chemistry And Biochemistry Of The Academy Of Sciences Of The Czech Republic Analogues de nucleotides enantiomeres antiretroviraux
EP0618214A1 (fr) * 1993-04-01 1994-10-05 Merrell Dow Pharmaceuticals Inc. Dérivés phosphonates insaturés de purines et de pyrimidines
EP0632048A1 (fr) * 1993-06-29 1995-01-04 Mitsubishi Chemical Corporation Dérivés esters phosphoniques de nucléotides
WO1996033200A1 (fr) * 1995-04-21 1996-10-24 Ústav Organické Chemie A Biochemie Akademie Ved C^¿Eské Republiky Nouveaux composes et procedes therapeutiques
EP0785208A1 (fr) * 1996-01-18 1997-07-23 Mitsubishi Chemical Corporation Composés phosphoniques de nucléotides
EP0832896A1 (fr) * 1995-06-15 1998-04-01 Mitsubishi Chemical Corporation Nucleotides derives de phosphonates
EP0919562A1 (fr) * 1996-08-13 1999-06-02 Mitsubishi Chemical Corporation Composes nucleotidiques a base de phosphonate
WO2001064693A1 (fr) * 2000-02-29 2001-09-07 Mitsubishi Pharma Corporation Compose de nucleotide de phosphonate

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994003467A2 (fr) * 1992-08-05 1994-02-17 Institute Of Organic Chemistry And Biochemistry Of The Academy Of Sciences Of The Czech Republic Analogues de nucleotides enantiomeres antiretroviraux
EP0618214A1 (fr) * 1993-04-01 1994-10-05 Merrell Dow Pharmaceuticals Inc. Dérivés phosphonates insaturés de purines et de pyrimidines
EP0632048A1 (fr) * 1993-06-29 1995-01-04 Mitsubishi Chemical Corporation Dérivés esters phosphoniques de nucléotides
US6037335A (en) * 1993-06-29 2000-03-14 Mitsubishi Chemical Corporation Phosphonate-nucleotide ester derivatives
WO1996033200A1 (fr) * 1995-04-21 1996-10-24 Ústav Organické Chemie A Biochemie Akademie Ved C^¿Eské Republiky Nouveaux composes et procedes therapeutiques
EP0832896A1 (fr) * 1995-06-15 1998-04-01 Mitsubishi Chemical Corporation Nucleotides derives de phosphonates
EP0785208A1 (fr) * 1996-01-18 1997-07-23 Mitsubishi Chemical Corporation Composés phosphoniques de nucléotides
EP0919562A1 (fr) * 1996-08-13 1999-06-02 Mitsubishi Chemical Corporation Composes nucleotidiques a base de phosphonate
WO2001064693A1 (fr) * 2000-02-29 2001-09-07 Mitsubishi Pharma Corporation Compose de nucleotide de phosphonate

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
COLACINO J M ET AL: "The identification and development of antiviral agents for the treatment of chronic hepatitis B virus infection.", PROGRESS IN DRUG RESEARCH. FORTSCHRITTE DER ARZNEIMITTELFORSCHUNG. PROGRES DES RECHERCHES PHARMACEUTIQUES. SWITZERLAND 1998, vol. 50, 1998, pages 259 - 322, XP000885900, ISSN: 0071-786X *
DATABASE WPI Section Ch Week 200165, Derwent World Patents Index; Class B02, AN 2001-582144, XP002221470 *
HU JIANMING ET AL: "Hepadnavirus assembly and reverse transcription require a multi-component chaperone complex which is incorporated into nucleocapsids.", EMBO (EUROPEAN MOLECULAR BIOLOGY ORGANIZATION) JOURNAL, vol. 16, no. 1, 1997, pages 59 - 68, XP002221467, ISSN: 0261-4189 *
SEKIYA, KOUICHI ET AL: "2-Amino-6-arylthio-9-[2-(phosphonomethoxy)ethyl]purine Bis(2,2,2-trifluoroethyl) Esters as Novel HBV-Specific Antiviral Reagents", JOURNAL OF MEDICINAL CHEMISTRY (2002), 45(14), 3138-3142, 6 April 2002 (2002-04-06), XP002221469 *
STASCHKE K A ET AL: "Drug discovery and development of antiviral agents for the treatment of chronic hepatitis B virus infection.", PROGRESS IN DRUG RESEARCH. FORTSCHRITTE DER ARZNEIMITTELFORSCHUNG. PROGRES DES RECHERCHES PHARMACEUTIQUES. SWITZERLAND 2001, vol. Spec No, 2001, pages 111 - 183, XP001121546, ISSN: 0071-786X *
STASCHKE K A ET AL: "Priming of duck hepatitis B virus reverse transcription in vitro: premature termination of primer DNA induced by the 5'-triphosphate of fialuridine.", JOURNAL OF VIROLOGY. UNITED STATES DEC 1994, vol. 68, no. 12, December 1994 (1994-12-01), pages 8265 - 8269, XP009001414, ISSN: 0022-538X *
TAVIS JOHN E ET AL: "Evidence for activation of the hepatitis B virus polymerase by binding of its RNA template.", JOURNAL OF VIROLOGY, vol. 70, no. 9, 1996, pages 5741 - 5750, XP002221466, ISSN: 0022-538X *
TAVIS JOHN E ET AL: "The duck hepatitis B virus polymerase is activated by its RNA packaging signal, epsilon.", JOURNAL OF VIROLOGY, vol. 72, no. 7, July 1998 (1998-07-01), pages 5789 - 5796, XP002221468, ISSN: 0022-538X *

Cited By (37)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7462608B2 (en) 2002-04-26 2008-12-09 Gilead Sciences, Inc. Non nucleoside reverse transcriptase inhibitors
US7649015B2 (en) 2002-04-26 2010-01-19 Gilead Sciences, Inc. Cellular accumulation of phosphonate analogs of HIV protease inhibitor compounds
WO2004046157A1 (fr) * 2002-11-15 2004-06-03 Mitsubishi Pharma Corporation Procede servant a preparer halogenure de 2-[bis(2,2,2-trifluoroethyl)phosphonylmethoxy]ethyl
US7470724B2 (en) 2003-04-25 2008-12-30 Gilead Sciences, Inc. Phosphonate compounds having immuno-modulatory activity
US7273716B2 (en) 2003-04-25 2007-09-25 Gilead Sciences, Inc. Methods and compositions for identifying therapeutic compounds with GS-7340 ester hydrolase
US9139604B2 (en) 2003-04-25 2015-09-22 Gilead Sciences, Inc. Antiviral phosphonate analogs
US8871785B2 (en) 2003-04-25 2014-10-28 Gilead Sciences, Inc. Antiviral phosphonate analogs
US7645747B2 (en) 2003-04-25 2010-01-12 Gilead Sciences, Inc. Therapeutic phosphonate compounds
US7407965B2 (en) 2003-04-25 2008-08-05 Gilead Sciences, Inc. Phosphonate analogs for treating metabolic diseases
US7417055B2 (en) 2003-04-25 2008-08-26 Gilead Sciences, Inc. Kinase inhibitory phosphonate analogs
WO2005002626A2 (fr) * 2003-04-25 2005-01-13 Gilead Sciences, Inc. Composes de phosphonate therapeutiques
US7427636B2 (en) 2003-04-25 2008-09-23 Gilead Sciences, Inc. Inosine monophosphate dehydrogenase inhibitory phosphonate compounds
EA014685B1 (ru) * 2003-04-25 2010-12-30 Джилид Сайэнс, Инк. Фосфонатсодержащие антивирусные соединения (варианты) и фармацевтическая композиция на их основе
US8022083B2 (en) 2003-04-25 2011-09-20 Gilead Sciences, Inc. Antiviral phosphonate analogs
US7432261B2 (en) 2003-04-25 2008-10-07 Gilead Sciences, Inc. Anti-inflammatory phosphonate compounds
US7429565B2 (en) 2003-04-25 2008-09-30 Gilead Sciences, Inc. Antiviral phosphonate analogs
US7452901B2 (en) 2003-04-25 2008-11-18 Gilead Sciences, Inc. Anti-cancer phosphonate analogs
WO2004096286A3 (fr) * 2003-04-25 2005-06-16 Gilead Sciences Inc Analogues de phosphonate antiviraux
WO2005002626A3 (fr) * 2003-04-25 2005-05-26 Gilead Sciences Inc Composes de phosphonate therapeutiques
US7300924B2 (en) 2003-04-25 2007-11-27 Gilead Sciences, Inc. Anti-infective phosphonate analogs
US7427624B2 (en) 2003-10-24 2008-09-23 Gilead Sciences, Inc. Purine nucleoside phosphorylase inhibitory phosphonate compounds
US7432273B2 (en) 2003-10-24 2008-10-07 Gilead Sciences, Inc. Phosphonate analogs of antimetabolites
US7273715B2 (en) 2003-10-24 2007-09-25 Gilead Sciences, Inc. Methods and compositions for identifying therapeutic compounds with GS-9005 ester hydrolase A
US7273717B2 (en) 2003-10-24 2007-09-25 Gilead Sciences, Inc. Methods and compositions for identifying therapeutic compounds with GS-9005 ester hydrolase B
KR101154532B1 (ko) 2003-11-12 2012-07-04 길리애드 사이언시즈, 인코포레이티드 항바이러스 포스포네이트 유사체
US7432272B2 (en) 2003-12-22 2008-10-07 Gilead Sciences, Inc. Antiviral analogs
US9457035B2 (en) 2004-07-27 2016-10-04 Gilead Sciences, Inc. Antiviral compounds
US9579332B2 (en) 2004-07-27 2017-02-28 Gilead Sciences, Inc. Phosphonate analogs of HIV inhibitor compounds
US8951986B2 (en) 2008-07-08 2015-02-10 Gilead Sciences, Inc. Salts of HIV inhibitor compounds
US9381206B2 (en) 2008-07-08 2016-07-05 Gilead Sciences, Inc. Salts of HIV inhibitor compounds
US9783568B2 (en) 2008-07-08 2017-10-10 Gilead Sciences, Inc. Salts of HIV inhibitor compounds
US9187507B2 (en) 2009-12-10 2015-11-17 Institute Of Pharmacology And Toxicology Academy Of Military Medical Sciences P.L.A. China Acyclic nucleoside phosphonate derivatives and medical uses thereof
US9593137B2 (en) 2011-12-22 2017-03-14 Geron Corporation Guanine analogs as telomerase substrates and telomere length affectors
US10035814B2 (en) 2011-12-22 2018-07-31 Geron Corporation Guanine analogs as telomerase substrates and telomere length affectors
US10562926B2 (en) 2011-12-22 2020-02-18 Geron Corporation Guanine analogs as telomerase substrates and telomere length affectors
US11279720B2 (en) 2011-12-22 2022-03-22 Geron Corporation Guanine analogs as telomerase substrates and telomere length affectors
US10851125B2 (en) 2017-08-01 2020-12-01 Gilead Sciences, Inc. Crystalline forms of ethyl ((S)-((((2R,5R)-5-(6-amino-9H-purin-9-yl)-4-fluoro-2,5-dihydrofuran-2-yl)oxy)methyl)(phenoxy)phosphoryl(-L-alaninate

Also Published As

Publication number Publication date
JP2005508924A (ja) 2005-04-07

Similar Documents

Publication Publication Date Title
WO2003028737A1 (fr) Agents antiviraux et procede in-vitro d'identification de candidats empechant la fixation de la polymerase a l'epsilon
US6767900B2 (en) Phosphonate nucleotide compound
CN107073005B (zh) 治疗丝状病毒科病毒感染的方法
JP4083691B2 (ja) 抗レトロウィルス性エナンチオマー性ヌクレオチドアナログ
AU2010319999B2 (en) 2'-Fluoro-6'-methylene carbocyclic nucleosides and methods of treating viral infections
TWI567074B (zh) ２’－氟－６’－亞甲基碳環核苷類及治療病毒感染之方法
KR100366727B1 (ko) 포스포네이트뉴클레오티드화합물
ES2392840T3 (es) Análogos de nucleótido para el tratamiento de infecciones virales
US5246924A (en) Method for treating hepatitis B virus infections using 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-ethyluracil
JP6860585B2 (ja) 非環状抗ウイルス 優先出願の参照による組み込み
Kreemerova Amino acid ester prodrugs of nucleoside and nucleotide antivirals
TW201036989A (en) Uracyl cyclopropyl nucleotides
KR19990022752A (ko) 포스포네이트 뉴클레오티드 유도체
AU2004309418B2 (en) 4'-substituted carbovir-and abacavir-derivatives as well as related compounds with HIV and HCV antiviral activity
JP3172801B2 (ja) 抗ウィルス剤としてのキラル２−（ホスホノメトキシ）プロピルグアニン
CA1293972C (fr) 1-¬2-(hydroxymethyl)cycloalkylmethyl|uraciles substituees en position 5
KR970011386B1 (ko) 1-(2'-데옥시-2'-플루오로-베타-d-아리비노-푸라노실)-5-에틸우라실을 이용한 간염 비루스 감염질환의 치료방법 및 치료용 조성물
US6914138B2 (en) Urea nucleosides as therapeutic and diagnostic agents
JP2020164521A (ja) 抗ウィルス薬
JP3561272B6 (ja) 抗レトロウィルス性エナンチオマー性ヌクレオチドアナログ
WO1993017035A1 (fr) 2'ISO-DIDESOXY-β-D-NUCLEOSIDES UTILES COMME AGENTS ANTIVIRAUX STABLES
JP2000503640A (ja) ヌクレオチドアナログ

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BY BZ CA CH CN CO CR CU CZ DE DM DZ EC EE ES FI GB GD GE GH HR HU ID IL IN IS JP KE KG KR KZ LK LR LS LT LU LV MA MD MG MK MW MX MZ NO NZ OM PH PL PT RO SD SE SG SI SK SL TJ TM TN TR TT UA UG US UZ VC VN YU ZA ZM

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW MZ SD SL SZ UG ZM ZW AM AZ BY KG KZ RU TJ TM AT BE BG CH CY CZ DK EE ES FI FR GB GR IE IT LU MC PT SE SK TR BF BJ CF CG CI GA GN GQ GW ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 2003532069

Country of ref document: JP

122 Ep: pct application non-entry in european phase