WO2020103929A1 - 低聚核苷酸及前体药物 - Google Patents

低聚核苷酸及前体药物

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Publication number
WO2020103929A1
WO2020103929A1 PCT/CN2019/120251 CN2019120251W WO2020103929A1 WO 2020103929 A1 WO2020103929 A1 WO 2020103929A1 CN 2019120251 W CN2019120251 W CN 2019120251W WO 2020103929 A1 WO2020103929 A1 WO 2020103929A1
Authority
WO
WIPO (PCT)
Prior art keywords
alkyl
pharmaceutically acceptable
compound
formula
alkoxy
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/CN2019/120251
Other languages
English (en)
French (fr)
Inventor
张寅生
刘保民
朱炎
黄雨
盖阔
秦慧
田晓萌
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Chia Tai Tianqing Pharmaceutical Group Co Ltd
Original Assignee
Chia Tai Tianqing Pharmaceutical Group Co Ltd
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 Chia Tai Tianqing Pharmaceutical Group Co Ltd filed Critical Chia Tai Tianqing Pharmaceutical Group Co Ltd
Priority to CN201980076695.2A priority Critical patent/CN113166190B/zh
Publication of WO2020103929A1 publication Critical patent/WO2020103929A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
    • A61K31/7072—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid having two oxo groups directly attached to the pyrimidine ring, e.g. uridine, uridylic acid, thymidine, zidovudine
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7076—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines containing purines, e.g. adenosine, adenylic acid
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7076—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines containing purines, e.g. adenosine, adenylic acid
    • A61K31/708—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines containing purines, e.g. adenosine, adenylic acid having oxo groups directly attached to the purine ring system, e.g. guanosine, guanylic acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/06—Pyrimidine radicals
    • C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/16—Purine radicals
    • C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • 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
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/29—Hepatitis virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • 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

Definitions

  • the present application relates to oligonucleotides and prodrugs, methods for their preparation, pharmaceutical compositions containing the compounds, and their use in the treatment of hepatitis B virus (HBV) infection and liver diseases related to HBV.
  • HBV hepatitis B virus
  • Chronic hepatitis B is currently incurable and can only be controlled.
  • Two anti-HBV drugs that have been clinically approved include interferon alpha and nucleoside analogs, but there is usually a rapid emergence of drug resistance and dose-related toxicity of treatment problem.
  • many current drugs suffer from poor absorption, distribution, metabolism, and / or excretion (ADME) properties, preventing their wider application. Poor ADME properties are also a major cause of failure of drug candidates in clinical trials.
  • ADME absorption, distribution, metabolism, and / or excretion
  • oligonucleotide drugs such as SB9000 and its prodrug SB9200 have both safety and antiviral effects in untreated chronic hepatitis B patients without cirrhosis, but there is still a need to develop better anti-HBV activity , Safety, prodrug release activity rate and bioavailability, higher stability and certain liver targeting compounds.
  • the present application relates to compounds of formula I, tautomers, stereoisomers or pharmaceutically acceptable salts thereof,
  • R 1 is selected from or Wherein R is selected from the following groups: C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 Aryl or 5-10 membered heteroaryl, the R group is optionally substituted with one or more of the following groups: halogen, -OH, C 1-6 alkyl, C 1-6 alkoxy,- NO 2 , -CN, -NH 2 , -NH (C 1-6 alkyl), -N (C 1-6 alkyl) 2 , -C (O) C 1-6 alkyl, -C (O) OC 1-6 alkyl, -CONHC 1-6 alkyl, -SH or -SC 1-6 alkyl;
  • R 2 and R 3 are independently selected from H, -OH, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl or 5-10 membered heteroaryl, the C 1-6 alkyl, C 1-6 alkoxy, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6 -10 aryl or 5-10 membered heteroaryl is optionally substituted with one or more substituents selected from halogen, -OH, -NO 2 , -NH 2 or C 1-6 alkoxy;
  • R 4 and R 5 are independently selected from H, -OH, -NH 2 or -SH;
  • n is selected from 0, 1, 2, 3 or 4;
  • R a and R b are independently selected from H, halogen, C 1-6 alkoxy or C 1-6 alkyl, the C 1-6 alkoxy or C 1-6 alkyl is optionally It is substituted by one or more substituents selected from C 1-6 alkoxy, halogen, -OH, -NH 2 or -NO 2 .
  • the present application relates to a pharmaceutical composition
  • a pharmaceutical composition comprising the compound of formula I of the present application, its tautomer, stereoisomer or a pharmaceutically acceptable salt thereof.
  • the present application relates to a method of treating or inhibiting HBV infection in a mammal, including administering to the mammal in need of such treatment a therapeutically effective amount of a compound of formula I, a tautomer, a stereoisomer, or a pharmaceutically acceptable Accepted salt or the above pharmaceutical composition.
  • the present application relates to the use of compounds of formula I, their tautomers, stereoisomers or pharmaceutically acceptable salts thereof or the above-mentioned pharmaceutical compositions in the preparation of a medicament for preventing or treating HBV infection in mammals.
  • the present application relates to the use of compounds of formula I, their tautomers, stereoisomers or pharmaceutically acceptable salts thereof or the above-mentioned pharmaceutical compositions in the prevention or treatment of mammalian HBV infection.
  • the present application relates to a compound of Formula I, its tautomer, stereoisomer or a pharmaceutically acceptable salt thereof or a pharmaceutical composition as described above for use in preventing or treating HBV infection in a mammal.
  • This application relates to compounds of formula I, their tautomers, stereoisomers or pharmaceutically acceptable salts thereof,
  • R 1 is selected from or Wherein R is selected from the following groups: C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 Aryl or 5-10 membered heteroaryl, the R group is optionally substituted with one or more groups selected from halogen, -OH, C 1-6 alkyl, C 1-6 alkoxy Group, -NO 2 , -CN, -NH 2 , -NH (C 1-6 alkyl), -N (C 1-6 alkyl) 2 , -C (O) C 1-6 alkyl, -C (O) OC 1-6 alkyl, -CONHC 1-6 alkyl, -SH or -SC 1-6 alkyl;
  • R 2 and R 3 are independently selected from H, -OH, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl or 5-10 membered heteroaryl, the C 1-6 alkyl, C 1-6 alkoxy, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6 -10 aryl or 5-10 membered heteroaryl is optionally substituted with one or more substituents selected from halogen, -OH, -NO 2 , -NH 2 or C 1-6 alkoxy;
  • R 4 and R 5 are independently selected from H, -OH, -NH 2 or -SH;
  • n is selected from 0, 1, 2, 3 or 4;
  • R a and R b are independently selected from H, halogen, C 1-6 alkoxy or C 1-6 alkyl, the C 1-6 alkoxy or C 1-6 alkyl is optionally It is substituted by one or more substituents selected from C 1-6 alkoxy, halogen, -OH, -NH 2 or -NO 2 .
  • R is selected from the following groups: C 1-6 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl , Phenyl, or 5-6 membered heteroaryl; in some embodiments, R is selected from C 1-4 alkyl; in some embodiments, R is isopropyl, methyl, or isobutyl.
  • R 2 and R 3 are each independently selected from H, —OH, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-10 cycloalkyl, 3-10 member Heterocycloalkyl, C 6-10 aryl or 5-10 membered heteroaryl, the C 1-6 alkyl or C 1-6 alkoxy group is optionally selected from one or more of halogen, -OH , -NO 2 , -NH 2 or C 1-6 alkoxy substituents; in some embodiments, R 2 and R 3 are each independently selected from H, halogen, C 1-6 alkyl or optional C 1-6 alkoxy substituted by C 1-6 alkoxy; in some embodiments, R 2 and R 3 are each independently selected from H, F, CH 3- , CH 3 O- or CH 3 OCH 2 CH 2 O-; in some embodiments, R 2 is H or F, and R 3 is CH 3- , CH 3 O-, or CH 3 OCH 2 CH 2 O-.
  • R 4 and R 5 are each independently selected from H or -OH; in some embodiments, R 4 is H and R 5 is -OH.
  • n is selected from 0, 1, or 2. In some embodiments, n is 1.
  • R a and R b are each independently selected from H, halogen, C 1-3 alkoxy, or C 1-3 alkyl; in some embodiments, R a and R b are each independently H or halogen.
  • the pharmaceutically acceptable salt is selected from sodium or ammonium salts.
  • the compound of formula I, its tautomer, stereoisomer or pharmaceutically acceptable salt thereof is selected from the compound of formula II, its tautomer, stereoisomer or Its pharmaceutically acceptable salts:
  • R 1 , R 2 or R 3 are as defined in the compound of formula I.
  • the pharmaceutically acceptable salts of the compounds of Formula I of the present application are selected from sodium salts or ammonium salts.
  • the compound of formula I of the present application is selected from:
  • the compound of formula I of the present application is selected from:
  • the present application relates to a pharmaceutical composition
  • a pharmaceutical composition comprising the compound of formula I of the present application, its tautomer, stereoisomer or a pharmaceutically acceptable salt thereof.
  • the pharmaceutical composition of the present application further comprises pharmaceutically acceptable excipients.
  • the present application relates to a method of treating or inhibiting HBV infection in mammals, including administering to a mammal in need of such treatment, preferably a human, a therapeutically effective amount of a compound of formula I, its tautomer, stereoisomer or A pharmaceutically acceptable salt, or the above pharmaceutical composition.
  • the present application relates to the use of the compound of formula I, its tautomer, stereoisomer or pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical composition in the preparation of a medicament for preventing or treating HBV infection in mammals .
  • the present application relates to the use of the compound of formula I, its tautomer, stereoisomer or pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical composition for preventing or treating HBV infection in mammals.
  • the present application relates to a compound of formula I for preventing or treating HBV infection in a mammal, its tautomer, stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above.
  • the treatment of HBV infection refers to the control, reduction or elimination of HBV to alleviate or cure liver disease in infected mammals or patients.
  • substituted means that any one or more hydrogen atoms on a specific group are substituted with a substituent, as long as the valence state of the specific group is normal and the compound after substitution is stable.
  • ethyl is "optionally” substituted with halogen, meaning that ethyl can be unsubstituted (CH 2 CH 3 ), mono-substituted (such as CH 2 CH 2 F), poly-substituted (such as CHFCH 2 F, CH 2 CHF 2 etc.) or completely substituted (CF 2 CF 3 ).
  • halogen meaning that ethyl can be unsubstituted (CH 2 CH 3 ), mono-substituted (such as CH 2 CH 2 F), poly-substituted (such as CHFCH 2 F, CH 2 CHF 2 etc.) or completely substituted (CF 2 CF 3 ).
  • C mn is an integer number of carbon atoms in the given range.
  • C 1-6 means that the group may have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms.
  • any variable (such as R) appears more than once in the composition or structure of a compound, its definition in each case is independent. So, for example, if a group is replaced by 2 Rs, then each R has an independent option.
  • halo or halogen refers to fluorine, chlorine, bromine and iodine.
  • alkyl refers to a hydrocarbon group of the formula C n H 2n +.
  • the alkyl group may be linear or branched.
  • C 1 - 6 alkyl refers to (e.g., methyl, ethyl, n-propyl, isopropyl, alkyl containing 1 to 6 carbon atoms, n-butyl, isobutyl, sec-butyl, Tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc.).
  • the alkyl portion (ie, alkyl) of alkoxy, alkylamino, dialkylamino, alkylsulfonyl, and alkylthio groups has the same definition as above.
  • alkoxy refers to -O-alkyl
  • alkenyl refers to a linear or branched unsaturated aliphatic hydrocarbon group composed of carbon atoms and hydrogen atoms and having at least one double bond.
  • alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, 1,3-butadienyl, and the like.
  • alkynyl refers to a linear or branched unsaturated aliphatic hydrocarbon group composed of carbon atoms and hydrogen atoms and having at least one triple bond.
  • alkynyl groups include, but are not limited to, ethynyl (-C ⁇ CH), 1-propynyl (-C ⁇ C-CH 3 ), 2-propynyl (-CH 2 -C ⁇ CH), 1,3-butadiynyl (-C ⁇ CC ⁇ CH) etc.
  • cycloalkyl refers to a carbocycle that is fully saturated and can exist in the form of a monocyclic, bridged or spiro ring. Unless otherwise indicated, the carbocyclic ring is usually a 3 to 10 membered ring or a 3 to 6 membered ring.
  • Non-limiting examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl (bicyclo [2.2.1] heptyl), bicyclo [2.2.2] octyl, adamant Alkyl and so on.
  • heterocycloalkyl refers to a cyclic group that is fully saturated and can exist in the form of a monocyclic, bridged ring, or spiro ring, containing 1 to 4 heteroatoms selected from N, O, and S. Unless otherwise indicated, the heterocyclic ring is usually a 3 to 7 membered ring or 3 to 10 membered ring containing 1 to 3 heteroatoms (preferably 1 or 2 heteroatoms) independently selected from sulfur, oxygen and / or nitrogen .
  • 3-membered heterocyclic alkyl groups include, but are not limited to, ethylene oxide, epithioethyl, and cyclic azetyl groups
  • 4-membered heterocyclic alkyl groups include, but are not limited to, azetidinyl, oxin
  • Examples of cyclic groups, thibutane, 5-membered heterocycloalkyl include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, thiazolidine
  • groups, imidazolidinyl, tetrahydropyrazolyl, and 6-membered heterocycloalkyl include, but are not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl
  • aryl refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated ⁇ electron system.
  • the aryl group may have 6-20 carbon atoms, 6-14 carbon atoms, 6-12 carbon atoms, or 6-10 carbon atoms.
  • Non-limiting examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, 1,2,3,4-tetrahydronaphthalene, and the like.
  • heteroaryl refers to a monocyclic or fused polycyclic ring system, which contains at least one (eg 1, 2, or 1 to 3) heteroatoms selected from N, O, and S as ring atoms, and the remaining rings
  • the atom is C and has at least one aromatic ring.
  • the heteroaryl group may be a 5- to 10-membered ring containing 1 to 3 heteroatoms independently selected from N, O, and S.
  • Preferred heteroaryl groups have a single 5- to 8-membered ring, or multiple fused rings containing 6 to 14, especially 6 to 10 ring atoms.
  • heteroaryl groups include, but are not limited to, pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolinyl, isoquinolinyl , Tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothienyl, indolyl, isoindolyl, etc.
  • preventing means preventing the occurrence of a disease or disease state in a mammal, especially when such mammals are susceptible to the disease state, but have not been diagnosed with the disease state.
  • treatment means administration of a compound or formulation described in this application to ameliorate or eliminate a disease or one or more symptoms associated with the disease, and includes:
  • terapéuticaally effective amount means (i) treatment or prevention of a specific disease, condition or disorder, (ii) reduction, improvement or elimination of one or more symptoms of a specific disease, condition or disorder, or (iii) prevention or delay
  • the amount of the compound of the present application constituting the “therapeutically effective amount” depends on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by those skilled in the art It is determined by its own knowledge and this disclosure.
  • pharmaceutically acceptable refers to those compounds, materials, compositions and / or dosage forms, which are within the scope of reliable medical judgment and are suitable for use in contact with human and animal tissues without Multiple toxicity, irritation, allergic reactions or other problems or complications are commensurate with a reasonable benefit / risk ratio.
  • Examples of pharmaceutically acceptable salts include metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like.
  • pharmaceutical composition refers to a mixture of one or more compounds of the present application or salts thereof and pharmaceutically acceptable excipients.
  • the purpose of the pharmaceutical composition is to facilitate the administration of the compound of the present application to an organism.
  • pharmaceutically acceptable excipients refers to those excipients that have no significant stimulating effect on the organism and do not impair the biological activity and performance of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water and the like.
  • tautomer or "tautomeric form” refers to structural isomers of different energies that can be interconverted via a low energy barrier.
  • proton tautomers also known as proton transfer tautomers
  • proton migration such as keto-enol and imine-enamine isomerization.
  • proton tautomers is an imidazole moiety, where protons can migrate between two ring nitrogens.
  • Valence tautomers include interconversion through the recombination of some bonding electrons.
  • the compounds of the present application may be asymmetric, for example, having one or more stereoisomers. Unless otherwise stated, all stereoisomers include, for example, enantiomers and diastereomers.
  • the compounds containing asymmetric carbon atoms of the present application can be isolated in optically active pure form or in racemic form. The optically active pure form can be resolved from the racemic mixture or synthesized by using chiral raw materials or chiral reagents.
  • the compounds of the present application may exist in specific geometric isomers or stereoisomeric forms. This application contemplates all such compounds, including tautomers, cis and trans isomers, (-)-and (+)-enantiomers, (R)-and (S) -enantiomers , Diastereomers, (D) -isomers, (L) -isomers, and their racemic mixtures and other mixtures, such as enantiomers or diastereomer-enriched mixtures, All of these are within the scope of this application. Additional asymmetric carbon atoms may be present in the substituents such as alkyl. All these isomers and their mixtures are included within the scope of this application.
  • the pharmaceutical composition of the present application can be prepared by combining the compound of the present application with suitable pharmaceutically acceptable excipients, for example, can be formulated into solid, semi-solid, liquid or gaseous formulations, such as tablets, pills, capsules, powders , Granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols.
  • suitable pharmaceutically acceptable excipients for example, can be formulated into solid, semi-solid, liquid or gaseous formulations, such as tablets, pills, capsules, powders , Granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols.
  • Typical routes for administering the compounds of the present application or pharmaceutically acceptable salts or pharmaceutical compositions thereof include, but are not limited to oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal Intramuscular, subcutaneous and intravenous administration.
  • the pharmaceutical composition of the present application can be manufactured by methods well known in the art, such as conventional mixing method, dissolution method, granulation method, sugar-coated pill method, grinding method, emulsification method, freeze-drying method, and the like.
  • the pharmaceutical composition is in oral form.
  • the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compound of the present application to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, etc. for oral administration to patients.
  • the solid oral composition can be prepared by conventional mixing, filling or tabletting methods. For example, it can be obtained by mixing the active compound with solid auxiliary materials, optionally grinding the resulting mixture, adding other suitable auxiliary materials if necessary, and then processing the mixture into granules to obtain tablets Or the core of sugar coating.
  • suitable excipients include but are not limited to: binders, diluents, disintegrants, lubricants, glidants, sweeteners, or flavoring agents.
  • the pharmaceutical composition may also be suitable for parenteral administration, such as a sterile solution, suspension or lyophilized product in a suitable unit dosage form.
  • the daily dose is 0.01 to 200 mg / kg body weight, either in separate or divided doses.
  • the compounds of the present application can be prepared by various synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by the combination with other chemical synthesis methods, and those well known to those skilled in the art Equivalently, preferred embodiments include but are not limited to the examples of the present application.
  • the compound of the general formula II of the present application can be prepared by a person skilled in the art of organic synthesis through the following route:
  • Bz stands for benzoyl
  • DMTr stands for 4,4'-dimethoxytriphenylmethyl
  • Me stands for methyl
  • TBDPS stands for tert-butyldiphenylsilyl
  • DMSO stands for dimethyl sulfoxide.
  • Example 1 O-(((1R, 3S, 5S) -3- (2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2- Methylenecyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl)- 2-hydroxymethyl-4-methoxytetrahydrofuran-3-yl) ammonium thiophosphate
  • Step 1 N- (9-((1S, 3R, 4S) -4-hydroxy-3-hydroxymethyl-2-methylenecyclopentyl) -6-oxo-6,9-dihydro-1H -Purin-2-yl) benzamide
  • reaction solution was cooled to 0 ° C in an ice bath, and water (50 ml) was slowly added, and stirred at 0 ° C for 30 min.
  • the ice bath was removed, and ammonia water (85 g) was added to the reaction solution, and stirred at room temperature for 30 min.
  • the reaction solution was concentrated under reduced pressure, and the resulting concentrate was added with water (50 ml) to beat. After filtration, the filter cake was washed successively with water and ether, and dried to obtain the title compound (5.8 g).
  • Step 2 (2R, 3R, 4R, 5R) -2-((bis (4-methoxyphenyl) (phenyl) methoxy) methyl) -5- (2,4-dioxo- 3,4-dihydropyrimidine-1 (2H) -yl) -4-methoxytetrahydrofuran-3-yl (2-cyanoethyl) diisopropylphosphoramidite
  • Step 3 ((1R, 3S, 5S) -3- (2-Benzamido-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2-methylene Methylcyclopentyl) methyl ((2R, 3R, 4R, 5R) -2-((bis (4-methoxyphenyl) (phenyl) methoxy) methyl) -5- (2, 4-dioxo-3,4-dihydropyrimidine-1 (2H) -yl) -4-methoxytetrahydrofuran-3-yl) (2-cyanoethyl) phosphite
  • Step 4 O-(((1R, 3S, 5S) -3- (2-benzoylamino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy- 2-methylenecyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -2-((bis (4-methoxyphenyl) (phenyl) methoxy) methyl) -5- (2,4-dioxo-3,4-dihydropyrimidine-1 (2H) -yl) -4-methoxytetrahydrofuran-3-yl) O- (2-cyanoethyl) sulfur Phosphoric acid ester
  • Step 5 O-(((1R, 3S, 5S) -3- (2-benzoylamino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy- 2-methylenecyclopentyl) methyl) O- (2-cyanoethyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-di Hydropyrimidine-1 (2H) -yl) -2-hydroxymethyl-4-methoxytetrahydrofuran-3-yl) phosphorothioate
  • Step 6 O-(((1R, 3S, 5S) -3- (2-Amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2-methylene Methylcyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) -2 -Hydroxymethyl-4-methoxytetrahydrofuran-3-yl) ammonium thiophosphate
  • Example 2 O-(((1R, 3S, 5S) -3- (2-amino-6-oxo-1H-purin-9 (6H) -yl) -5-hydroxy-2-methylene ring Amyl) methyl) O-((2R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine-1 (2H) -yl) -2-hydroxymethyl- 4-methoxytetrahydrofuran-3-yl) (S) -sodium thiophosphate
  • Step 1 (2S, 3aS, 6R, 7aS) -3a-methyl-2-((perfluorophenyl) thio) -6- (prop-1-en-2-yl) hexahydrobenzo [d ] [1,3,2] oxathiophosphorane 2-sulfide
  • reaction solution was concentrated to 500 ml, 1000 ml of n-hexane was added to the reaction solution for dilution, and then washed with 10% potassium dihydrogen phosphate solution and saturated saline, respectively.
  • the organic phase was concentrated to 200 ml, 200 ml of methanol was added, and after further concentration to 50 ml, 200 ml of methanol was added, and then concentrated to 50 ml, and a solid precipitated. After cooling to 0 ° C in an ice bath, it was filtered.
  • the crude product was 93.48g.
  • the crude product (93.48g) was dissolved in 225ml of dichloromethane, and then 374ml of n-heptane was added, and the crude product was completely dissolved.
  • the solution was concentrated to 200 ml, and then 374 ml of n-heptane was added, and this operation was repeated 3 times. After the solution was concentrated to 200 ml, a large amount of solid precipitated, and the mixture was stirred at room temperature for 2 h. Filter and dry the filter cake to give the title compound (62.58g).
  • Step 2 1-((2R, 3R, 4R, 5R) -5-(((tert-butyldiphenylsilyl) oxy) methyl) -3-methoxy-4-(((2R, 3aS, 6R, 7aS) -3a-methyl-6- (prop-1-en-2-yl) -2-sulfonylhexahydrobenzo [d] [1,3,2] oxathiophosphorus heterocycle Pentane-2-oxy) oxy) tetrahydrofuran-2-yl) pyrimidine-2,4 (1H, 3H))-dione
  • Step 3 O-(((1R, 3S, 5S) -3- (2-benzoyl-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-((tert Butyldiphenylsilyl) oxy) -2-methylenecyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -2-(((tert-butyldiphenylsilane Yl) oxy) methyl) -5- (2,4-dioxo-3,4-dihydropyrimidine-1) (2H) -yl) -4-methoxytetrahydrofuran-3-yl) O- Hydrogen (S)-phosphorothioate
  • Step 4 O-(((1R, 3S, 5S) -3- (2-benzoylamino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy- 2-methylenecyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -2-hydroxymethyl-5- (2,4-dioxo-3,4-dihydropyrimidine- 1 (2H) -yl) -4-methoxytetrahydrofuran-3-yl) (S) -tetrabutylammonium thiophosphate
  • Step 5 O-(((1R, 3S, 5S) -3- (2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2- Methylcyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) -2 -Hydroxymethyl-4-methoxytetrahydrofuran-3-yl) (S) -Tetrabutylammonium thiophosphate
  • Step 6 O-(((1R, 3S, 5S) -3- (2-Amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2-methylene Methylcyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) -2 -Hydroxymethyl-4-methoxytetrahydrofuran-3-yl) (S) -Sodium thiophosphate
  • Example 3 O-(((1R, 3S, 5S) -3- (2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2- Methylenecyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl)- 2-hydroxymethyl-4- (2-methoxyethoxy) tetrahydrofuran-3-yl) (S) -sodium phosphorothioate
  • Step 1 1-((2R, 3R, 4R, 5R) -5-(((tert-butyldiphenylsilyl) oxy) methyl) -3- (2-methoxyethoxy)- 4-(((2R, 3aS, 6R, 7aS) -3a-methyl-6- (prop-1-en-2-yl) -2-sulfonylhexahydrobenzo [d] [1,3,2 ] Oxathiophosphoran-2-yl) oxy) tetrahydrofuran-2-yl) pyrimidine-2,4- (1H, 3H) -dione
  • Step 2 O-(((1R, 3S, 5S) -3- (2-benzoyl-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-((tert Butyldiphenylsilyl) oxy) -2-methylenecyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -2-(((tert-butyldiphenylsilane Yl) oxy) methyl) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) -4- (2-methoxyethoxy) tetrahydrofuran- 3-yl) O-hydrogen (S) -phosphorothioate
  • Step 3 O-(((1R, 3S, 5S) -3- (2-benzoylamino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy- 2-methylenecyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl ) -2-hydroxymethyl-4- (2-methoxyethoxy) tetrahydrofuran-3-yl) (S) -tetrabutylammonium thiophosphate
  • Step 4 O-(((1R, 3S, 5S) -3- (2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2- Methylcyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) -2 -Hydroxymethyl-4- (2-methoxyethoxy) tetrahydrofuran-3-yl) (S) -tetrabutylammonium thiophosphate
  • Step 5 O-(((1R, 3S, 5S) -3- (2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2- Methylcyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) -2 -Hydroxymethyl-4- (2-methoxyethoxy) tetrahydrofuran-3-yl) (S) -sodium thiophosphate
  • Example 4 O-(((1R, 3S, 5S) -3- (2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2- Methylenecyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) 4-fluoro-2-hydroxymethyl-4-methyltetrahydrofuran-3-yl) (S) -sodium phosphorothioate
  • Step 1 1-((2R, 3R, 4R, 5R) -5-(((tert-butyldiphenylsilyl) oxy) methyl) -3-fluoro-4-hydroxy-3-methyltetrahydrofuran -2-yl) pyrimidine-2,4 (1H, 3H) -dione
  • Step 2 1-((2R, 3R, 4R, 5R) -5-(((tert-butyldiphenylsilyl) oxy) methyl) -3-fluoro-3-methyl-4-(( (2R, 3aS, 6R, 7aS) -3a-methyl-6- (prop-1-en-2-yl) -2-thiohexahydrobenzo [d] [1,3,2] oxathia Phosphalolan-2-yl) oxy) tetrahydrofuran-2-yl) pyrimidine-2,4 (1H, 3H) -dione
  • reaction solution was diluted with ethyl acetate and washed with 10% potassium dihydrogen phosphate solution.
  • Step 3 O-(((1R, 3S, 5S) -3- (2-benzoyl-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-((tert Butyldiphenylsilyl) oxy) -2-methylenecyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -2-(((tert-butyldiphenylsilane Yl) oxy) methyl) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) -4-fluoro-4-methyltetrahydrofuran-3-yl) O-hydrogen (S) -phosphorothioate
  • reaction solution was diluted with ethyl acetate and washed with saturated sodium bicarbonate solution.
  • Step 4 O-(((1R, 3S, 5S) -3- (2-benzoylamino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy- 2-methylenecyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl ) -4-fluoro-2-hydroxymethyl-4-methyltetrahydrofuran-3-yl) (S) -tetrabutylammonium thiophosphate
  • Step 5 O-(((1R, 3S, 5S) -3- (2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2- Methylcyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) -4 -Fluoro-2-hydroxymethyl-4-methyltetrahydrofuran-3-yl) (S) -tetrabutylammonium thiophosphate
  • Step 6 O-(((1R, 3S, 5S) -3- (2-Amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2-methylene Methylcyclopentyl) methyl) O-((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) -4 -Fluoro-2-hydroxymethyl-4-methyltetrahydrofuran-3-yl) (S) -sodium thiophosphate
  • Example 5 ((((((1R, 3S, 5S) -3- (2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5-hydroxy-2- Methylenecyclopentyl) methoxy) (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl)- 2-hydroxymethyl-4-methoxytetrahydrofuran-3-yl) oxy) phosphoryl) thio) methylisopropyl carbonate
  • Example 6 ((((S)-(((1R, 3S, 5S) -3- (2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5- Hydroxy-2-methylenecyclopentyl) methoxy) (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine-1 (2H) -Yl) -2-hydroxymethyl-4-methoxytetrahydrofuran-3-yl) oxy) phosphoryl) thio) methylisopropyl carbonate
  • Example 7 ((((S)-(((1R, 3S, 5S) -3- (2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5- Hydroxy-2-methylenecyclopentyl) methoxy) (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine-1 (2H) -Yl) -2-hydroxymethyl-4-methoxytetrahydrofuran-3-yl) oxy) phosphoryl) thio) methylisobutyl carbonate
  • Example 8 (((S)-(((1R, 3S, 5S) -3- (2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl) -5- Hydroxy-2-methylenecyclopentyl) methoxy) (((2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydropyrimidine-1 (2H) -Yl) -2-hydroxymethyl-4-methoxytetrahydrofuran-3-yl) oxy) phosphoryl) thio) methyl methyl carbonate
  • Test compound The compound prepared in the example was formulated into 20 mM mother liquor with dimethyl sulfoxide (DMSO).
  • the reagents used in this experiment include QIAamp 96 DNA Kit (12) (Cat. No. Qiagen-51162), FastStart Universal Probe (Cat. No. Roche-04914058001), CellTiter-Blue detection reagent (Cat. No. Promega-G808B)
  • the concentration of the test compound is always 100 ⁇ M, 3 times dilution, 8 concentrations.
  • Inhibition rate% (1-HBV DNA content in the sample / HBV DNA content in the DMSO control group) x 100%.
  • Cell viability% (Fluorescence value of sample well-Fluorescence value of medium control) / (Fluorescence value of DMSO control group-Fluorescence value of medium control) ⁇ 100%.
  • Example compounds EC 50 (nM) CC 50 ( ⁇ M) Example 1 441 > 100

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Abstract

公开了一种低聚核苷酸及前体药物，具体而言，公开了式I所示的化合物、其互变异构体、立体异构体或其药学上可接受的盐、制备方法、药物组合物及医药用途，包括用于治疗乙型肝炎病毒感染的用途，尤其是乙型肝炎病毒感染引起的肝脏疾病。

Description

低聚核苷酸及前体药物
相关申请的交叉引用
本申请要求于2018年11月23日向中国国家知识产权局提交的第201811408257.5号中国专利申请的优先权和权益，所述申请公开的内容通过引用整体并入本文中。
技术领域
本申请涉及低聚核苷酸及前体药物、其制备方法、含有该化合物的药物组合物、以及其在治疗乙型肝炎病毒(HBV)感染和与HBV有关的肝脏疾病方面的用途。
背景技术
慢性乙型病毒性肝炎目前不可治愈只能控制，临床中已经认可的两种抗HBV药物包括α干扰素和核苷类似物，但是通常存在抗药性的快速出现和与治疗有关的剂量限制的毒性问题。另外很多当前的药物受到吸收、分布、代谢和/或排泄(ADME)特性差的困扰，妨碍了其更广泛的应用。ADME特性差也是候选药物在临床试验中失败的一大原因。
已有的低聚核苷酸药物如SB9000及其前体药SB9200，在未经治疗无肝硬化的慢性乙肝患者中兼具安全性和抗病毒效用，但是仍需要开发具有更优的抗HBV活性、安全性、前药释放活性速度及生物利用率，较高的稳定性及一定的肝靶向性的化合物。
发明概述
一方面，本申请涉及式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，
Figure PCTCN2019120251-appb-000001
R 1选自
Figure PCTCN2019120251-appb-000002
或
Figure PCTCN2019120251-appb-000003
其中R选自以下基团：C 1-6烷基、C 2-6烯基、C 2-6炔基、C 3-6 环烷基、3-6元杂环烷基、C 6-10芳基或5-10元杂芳基，所述R基团任选地被一个或多个以下基团取代：卤素、-OH、C 1-6烷基、C 1-6烷氧基、-NO 2、-CN、-NH 2、-NH(C 1-6烷基)、-N(C 1-6烷基) 2、-C(O)C 1-6烷基、-C(O)OC 1-6烷基、-CONHC 1-6烷基、-SH或-SC 1-6烷基；
R 2及R 3分别独立地选自H、-OH、卤素、C 1-6烷基、C 1-6烷氧基、C 3-10环烷基、3-10元杂环烷基、C 6-10芳基或5-10元杂芳基，所述C 1-6烷基、C 1-6烷氧基、C 3-10环烷基、3-10元杂环烷基、C 6-10芳基或5-10元杂芳基任选地被一个或多个选自卤素、-OH、-NO 2、-NH 2或C 1-6烷氧基的取代基取代；
R 4及R 5分别独立地选自H、-OH、-NH 2或-SH；
n选自0、1、2、3或4；
X选自-C(R aR b)-或-C(＝CH 2)-；
其中，R a和R b分别独立地选自H、卤素、C 1-6烷氧基或C 1-6烷基，所述C 1-6烷氧基或C 1-6烷基任选地被一个或多个选自C 1-6烷氧基、卤素、-OH、-NH 2或-NO 2的取代基取代。
另一方面，本申请涉及药物组合物，其包含本申请的式I化合物、其互变异构体、立体异构体或其药学上可接受的盐。
另一方面，本申请涉及治疗或抑制哺乳动物HBV感染的方法，包括对需要该治疗的哺乳动物给予治疗有效量的式I化合物、其互变异构体、立体异构体或其药学上可接受的盐或上述药物组合物。
另一方面，本申请涉及式I化合物、其互变异构体、立体异构体或其药学上可接受的盐或上述药物组合物在制备预防或者治疗哺乳动物HBV感染的药物中的用途。
另一方面，本申请涉及式I化合物、其互变异构体、立体异构体或其药学上可接受的盐或上述药物组合物在预防或者治疗哺乳动物HBV感染中的用途。
另一方面，本申请涉及用于预防或者治疗哺乳动物HBV感染的式I化合物、其互变异构体、立体异构体或其药学上可接受的盐或上述药物组合物。
发明详述
本申请涉及式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，
Figure PCTCN2019120251-appb-000004
R 1选自
Figure PCTCN2019120251-appb-000005
或
Figure PCTCN2019120251-appb-000006
其中R选自以下基团：C 1-6烷基、C 2-6烯基、C 2-6炔基、C 3-6环烷基、3-6元杂环烷基、C 6-10芳基或5-10元杂芳基，所述R基团任选地被一个或多个选自以下的基团取代：卤素、-OH、C 1-6烷基、C 1-6烷氧基、-NO 2、-CN、-NH 2、-NH(C 1-6烷基)、-N(C 1-6烷基) 2、-C(O)C 1-6烷基、-C(O)OC 1-6烷基、-CONHC 1-6烷基、-SH或-SC 1-6烷基；
R 2及R 3分别独立地选自H、-OH、卤素、C 1-6烷基、C 1-6烷氧基、C 3-10环烷基、3-10元杂环烷基、C 6-10芳基或5-10元杂芳基，所述C 1-6烷基、C 1-6烷氧基、C 3-10环烷基、3-10元杂环烷基、C 6-10芳基或5-10元杂芳基任选地被一个或多个选自卤素、-OH、-NO 2、-NH 2或C 1-6烷氧基的取代基取代；
R 4及R 5分别独立地选自H、-OH、-NH 2或-SH；
n选自0、1、2、3或4；
X选自-C(R aR b)-或-C(＝CH 2)-；
其中，R a和R b分别独立地选自H、卤素、C 1-6烷氧基或C 1-6烷基，所述C 1-6烷氧基或C 1-6烷基任选地被一个或多个选自C 1-6烷氧基、卤素、-OH、-NH 2或-NO 2的取代基取代。
在一些实施方案中，
Figure PCTCN2019120251-appb-000007
与
Figure PCTCN2019120251-appb-000008
结合，形成
Figure PCTCN2019120251-appb-000009
其中
Figure PCTCN2019120251-appb-000010
为一价阳离子。在一些实施方案中，
Figure PCTCN2019120251-appb-000011
为Na +或NH 4 +。
在一些实施方案中，R选自以下基团：C 1-6烷基、C 2-4烯基、C 2-4炔基、C 3-6环烷基、3-6元杂环烷基、苯基或5-6元杂芳基；在一些实施方案中，R选自C 1-4烷基；在一些实施方案中，R为异丙基、甲基或异丁基。
在一些实施方案中，R 2及R 3分别独立地选自H、-OH、卤素、C 1-6烷基、C 1-6烷氧基、C 3-10环烷基、3-10元杂环烷基、C 6-10芳基或5-10元杂芳基，所述C 1-6烷基或C 1-6烷氧基任选地被一个或多个选自卤素、-OH、-NO 2、-NH 2或C 1-6烷氧基的取代基取代；在一些实施方案中，R 2及R 3分别独立地选自H、卤素、C 1-6烷基或任选地被C 1-6烷氧基取代的C 1-6烷氧基； 在一些实施方案中，R 2及R 3分别独立地选自H、F、CH 3-、CH 3O-或CH 3OCH 2CH 2O-；在一些实施方案中，R 2为H或F，R 3为CH 3-、CH 3O-或CH 3OCH 2CH 2O-。
在一些实施方案中，R 4及R 5分别独立地选自H或-OH；在一些实施方案中，R 4为H，R 5为-OH。
在一些实施方案中，n选自0、1或2。在一些实施方案中，n为1。
在一些实施方案中，R a和R b分别独立地选自H、卤素、C 1-3烷氧基或C 1-3烷基；在一些实施方案中，R a和R b分别独立地为H或卤素。
在一些实施方案中，X为-C(＝CH 2)-。
在一些实施方案中，药学上可接受的盐选自钠盐或铵盐。
在一些实施方案中，本申请的式I化合物、其互变异构体、立体异构体或其药学上可接受的盐选自式II化合物、其互变异构体、立体异构体或其药学上可接受的盐：
Figure PCTCN2019120251-appb-000012
其中，R 1、R 2或R 3同式I化合物中的定义。
在一些实施方案中，本申请式I化合物的药学上可接受的盐选自钠盐或铵盐。
在一些实施方案中，本申请的式I化合物、其互变异构体、立体异构体或其药学上可接受的盐选自：
Figure PCTCN2019120251-appb-000013
Figure PCTCN2019120251-appb-000014
在一些实施方案中，本申请的式I化合物、其互变异构体、立体异构体或其药学上可接受的盐选自：
Figure PCTCN2019120251-appb-000015
另一方面，本申请涉及药物组合物，其包含本申请的式I化合物、其互变异构体、立体异构体或其药学上可接受的盐。在一些实施方案中，本申请的药物组合物还包含药学上可接受的辅料。
另一方面，本申请涉及治疗或抑制哺乳动物HBV感染方法，包括对需要该治疗的哺乳动物，优选人类，给予治疗有效量的式I化合物、其互变异构体、立体异构体或其药学上可接受的盐、或上述药物组合物。
另一方面，本申请涉及式I化合物、其互变异构体、立体异构体或其药学上可接受的盐、或上述药物组合物在制备预防或者治疗哺乳动物HBV感染的药物中的用途。
另一方面，本申请涉及式I化合物、其互变异构体、立体异构体或其药学上可接受的盐、或上述药物组合物在预防或者治疗哺乳动物HBV感染的用途。
另一方面，本申请涉及预防或者治疗哺乳动物HBV感染的式I化合物、其互变异构体、立体异构体或其药学上可接受的盐、或上述药物组合物。
在本申请的部分实施方式中，所述治疗HBV感染指控制、降低或清除HBV以缓解或治愈受感染哺乳动物或患者的肝脏疾病。
定义
除非另有说明，本申请中所用的下列术语具有下列含义。一个特定的术语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照本领域普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。
术语“被取代”是指特定基团上的任意一个或多个氢原子被取代基取代，只要特定基团的价态是正常的并且取代后的化合物是稳定的。
术语“任选”或“任选地”是指随后描述的事件或情况可以发生或不发生，该描述包括发生所述事件或情况和不发生所述事件或情况。例如，乙基“任选”被卤素取代，指乙基可以是未被取代的(CH 2CH 3)、单取代的(如CH 2CH 2F)、多取代的(如CHFCH 2F、CH 2CHF 2等)或完全被取代的(CF 2CF 3)。本领域技术人员可理解，对于包含一个或多个取代基的任何基团，不会引入任何在空间上不可能存在和/或不能合成的取代或取代模式。
本文中的C m-n，是该部分具有给定范围中的整数个碳原子。例如“C 1-6”是指该基团可具有1个碳原子、2个碳原子、3个碳原子、4个碳原子、5个碳原子或6个碳原子。
当任何变量(例如R)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。因此，例如，如果一个基团被2个R所取代，则每个R都有独立的选项。
术语“卤”或“卤素”是指氟、氯、溴和碘。
术语“烷基”是指通式为C nH 2n+1的烃基。该烷基可以是直链或支链的。例如，术语“C 1- 6 烷基”指含有1至6个碳原子的烷基(例如甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、1-甲基丁基、2-甲基丁基、3-甲基丁基、新戊基、己基、2-甲基戊基等)。类似地，烷氧基、烷基氨基、二烷基氨基、烷基磺酰基和烷硫基的烷基部分(即烷基)具有上述相同定义。
术语“烷氧基”指-O-烷基。
术语“烯基”是指由碳原子和氢原子组成的直链或支链的具有至少一个双键的不饱和脂肪族烃基。烯基的非限制性实例包括但不限于乙烯基、1-丙烯基、2-丙烯基、1-丁烯基、异丁烯基、1,3-丁二烯基等。
术语“炔基”是指由碳原子和氢原子组成的直链或支链的具有至少一个三键的不饱和脂肪族烃基。炔基的非限制性实例包括但不限于乙炔基(-C≡CH)、1-丙炔基(-C≡C-CH 3)、2-丙炔基(-CH 2-C≡CH)、1,3-丁二炔基(-C≡C-C≡CH)等。
术语“环烷基”指完全饱和的并且可以以呈单环、桥环或螺环形式存在的碳环。除非另有指示，该碳环通常为3至10元环或3至6元环。环烷基非限制性实例包括但不限于环丙基、环丁基、环戊基、环己基、降冰片基(双环[2.2.1]庚基)、双环[2.2.2]辛基、金刚烷基等。
术语“杂环烷基”是指完全饱和的并且可以以单环、桥环或螺环形式存在的环状基团，其含有1至4个选自N、O和S的杂原子。除非另有指示，该杂环通常为含有1至3个独立地选自硫、氧和/或氮的杂原子(优选1或2个杂原子)的3至7元环或3至10元环。3元杂环烷基的实例包括但不限于环氧乙烷基、环硫乙烷基、环氮乙烷基，4元杂环烷基的非限制性实例包括但不限于吖丁啶基、噁丁环基、噻丁环基，5元杂环烷基的实例包括但不限于四氢呋喃基、四氢噻吩基、吡咯烷基、异噁唑烷基、噁唑烷基、异噻唑烷基、噻唑烷基、咪唑烷基、四氢吡唑基，6元杂环烷基的实例包括但不限于哌啶基、四氢吡喃基、四氢噻喃基、吗啉基、哌嗪基、1,4-噻噁烷基、1,4-二氧六环基、硫代吗啉基、1,3-二噻烷基、1,4-二噻烷基，7元杂环烷基的实例包括但不限于氮杂环庚烷基、氧杂环庚烷基、硫杂环庚烷基。优选为具有5或6个环原子的单环杂环烷基。
术语“芳基”是指具有共轭的π电子体系的全碳单环或稠合多环的芳香环基团。例如，芳基可以具有6-20个碳原子，6-14个碳原子、6-12个碳原子或6-10个碳原子。芳基的非限制性实例包括但不限于苯基、萘基、蒽基和1,2,3,4-四氢化萘等。
术语“杂芳基”是指单环或稠合多环体系，其中含有至少一个(例如1个、2个或1至3个)选自N、O、S的杂原子作为环原子，其余环原子为C，并且具有至少一个芳香环。杂芳基可以为含有1至3个独立地选自N、O和S的杂原子的5至10元环。优选的杂芳基具有单个5至8元环，或包含6至14个，尤其是6至10个环原子的多个稠合环。杂芳基的非限制 性实例包括但不限于吡咯基、呋喃基、噻吩基、咪唑基、噁唑基、吡唑基、吡啶基、嘧啶基、吡嗪基、喹啉基、异喹啉基、四唑基、三唑基、三嗪基、苯并呋喃基、苯并噻吩基、吲哚基、异吲哚基等。
术语“预防”意为预防疾病或疾病状态在哺乳动物中出现，特别是当这类哺乳动物易患有该疾病状态，但尚未被诊断为已患有该疾病状态时。
术语“治疗”意为将本申请所述化合物或制剂进行给药以改善或消除疾病或与所述疾病相关的一个或多个症状，且包括：
(i)抑制疾病或疾病状态，即遏制其发展；
(i)缓解疾病或疾病状态，即使该疾病或疾病状态消退。
术语“治疗有效量”意指(i)治疗或预防特定疾病、病况或障碍，(ii)减轻、改善或消除特定疾病、病况或障碍的一种或多种症状，或(iii)预防或延迟本文中所述的特定疾病、病况或障碍的一种或多种症状发作的本申请化合物的用量。构成“治疗有效量”的本申请化合物的量取决于该化合物、疾病状态及其严重性、给药方式以及待被治疗的哺乳动物的年龄而改变，但可例行性地由本领域技术人员根据其自身的知识及本公开内容而确定。
术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。
作为药学上可接受的盐，例如，可以为金属盐、铵盐、与有机碱形成的盐、与无机酸形成的盐、与有机酸形成的盐、与碱性或者酸性氨基酸形成的盐等。
术语“药物组合物”是指一种或多种本申请的化合物或其盐与药学上可接受的辅料组成的混合物。药物组合物的目的是有利于对有机体给予本申请的化合物。
术语“药学上可接受的辅料”是指对有机体无明显刺激作用，而且不会损害该活性化合物的生物活性及性能的那些辅料。合适的辅料是本领域技术人员熟知的，例如碳水化合物、蜡、水溶性和/或水可膨胀的聚合物、亲水性或疏水性材料、明胶、油、溶剂、水等。
词语“包括(comprise)”或“包含(comprise)”及其英文变体例如comprises或comprising应理解为开放的、非排他性的意义，即“包括但不限于”。
本申请的化合物和中间体还可以以不同的互变异构体形式存在，并且所有这样的形式包含于本申请的范围内。术语“互变异构体”或“互变异构体形式”是指可经由低能垒互变的不同能量的结构异构体。例如，质子互变异构体(也称为质子转移互变异构体)包括经由质子迁移的互变，如酮-烯醇及亚胺-烯胺异构化。质子互变异构体的具体实例是咪唑部分，其中质子可在两个环氮间迁移。价互变异构体包括通过一些成键电子的重组的互变。
本申请化合物可以是不对称的，例如，具有一个或多个立体异构体。除非另有说明，所有立体异构体都包括，如对映异构体和非对映异构体。本申请的含有不对称碳原子的化合物可以以光学活性纯的形式或外消旋形式被分离出来。光学活性纯的形式可以从外消旋混合物拆分，或通过使用手性原料或手性试剂合成。
本申请的化合物可以存在特定的几何异构体或立体异构体形式。本申请设想所有的这类化合物，包括互变异构体、顺式和反式异构体、(-)-和(+)-对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体富集的混合物，所有这些都属于本申请的范围之内。烷基等取代基中可以存在另外的不对称碳原子。所有这些异构体以及它们的混合物均包括在本申请的范围之内。
本申请的药物组合物可通过将本申请的化合物与适宜的药学上可接受的辅料组合而制备，例如可配制成固态、半固态、液态或气态制剂，如片剂、丸剂、胶囊剂、粉剂、颗粒剂、膏剂、乳剂、悬浮剂、栓剂、注射剂、吸入剂、凝胶剂、微球及气溶胶等。
给予本申请化合物或其药学上可接受的盐或其药物组合物的典型途径包括但不限于口服、直肠、局部、吸入、肠胃外、舌下、阴道内、鼻内、眼内、腹膜内、肌内、皮下、静脉内给药。
本申请的药物组合物可以采用本领域众所周知的方法制造，如常规的混合法、溶解法、制粒法、制糖衣药丸法、磨细法、乳化法、冷冻干燥法等。
在一些实施方案中，药物组合物是口服形式。对于口服给药，可以通过将活性化合物与本领域熟知的药学上可接受的辅料混合，来配制该药物组合物。这些辅料能使本申请的化合物被配制成片剂、丸剂、锭剂、糖衣剂、胶囊剂、液体、凝胶剂、浆剂、悬浮剂等，用于对患者的口服给药。
可以通过常规的混合、填充或压片方法来制备固体口服组合物。例如，可通过下述方法获得：将所述的活性化合物与固体辅料混合，任选地碾磨所得的混合物，如果需要则加入其它合适的辅料，然后将该混合物加工成颗粒，得到了片剂或糖衣剂的核心。适合的辅料包括但不限于：粘合剂、稀释剂、崩解剂、润滑剂、助流剂、甜味剂或矫味剂等。
药物组合物还可适用于肠胃外给药，如合适的单位剂型的无菌溶液剂、混悬剂或冻干产品。
本文所述的通式Ⅰ化合物的所有施用方法中，每天给药的剂量为0.01到200mg/kg体重，以单独或分开剂量的形式。
本申请的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所 熟知的等同替换方式，优选的实施方式包括但不限于本申请的实施例。
本申请具体实施方式的化学反应是在合适的溶剂中完成的，所述的溶剂须适合于本申请的化学变化及其所需的试剂和物料。为了获得本申请的化合物，有时需要本领域技术人员在已有实施方式的基础上对合成步骤或者反应流程进行修改或选择。
本领域合成路线规划中的一个重要考量因素是为反应性官能团(如本申请中的羟基)选择合适的保护基，例如，可参考Greene's Protective Groups in Organic Synthesis(4th Ed).Hoboken,New Jersey:John Wiley & Sons,Inc.本申请引用的所有参考文献整体上并入本申请。
在一些实施方案中，本申请通式II的化合物可以由有机合成领域技术人员通过以下路线来制备：
路线1：
Figure PCTCN2019120251-appb-000016
Figure PCTCN2019120251-appb-000017
本申请采用下述缩略词：
Bz代表苯甲酰基；DMTr代表4,4'-二甲氧基三苯基甲基；Me代表甲基；TBDPS代表叔丁基二苯基硅烷基；DMSO代表二甲基亚砜。
为清楚起见，进一步用实施例来阐述本发明，但是实施例并非限制本申请的范围。本申请所使用的所有试剂是市售的，无需进一步纯化即可使用。
具体实施方案
实施例1：O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)硫代磷酸铵
步骤1：N-(9-((1S，3R，4S)-4-羟基-3-羟甲基-2-亚甲基环戊基)-6-氧代-6,9-二氢-1H-嘌呤-2-基)苯甲酰胺
Figure PCTCN2019120251-appb-000018
将2-氨基-9-((1S,3R,4S)-4-羟基-3-羟甲基-2-亚甲基环戊基)-1,9-二氢-6H-嘌呤-6-酮(5.0g)与吡啶(80ml)混合。冰浴降温，将三甲基氯硅烷(14.69g)滴加至其中，滴毕室温反应3h。冰浴降温，缓慢滴加苯甲酰氯(4.31g)，加毕室温反应过夜。反应液冰浴降温至0℃，缓慢加入水(50ml)，0℃下搅拌30min。移去冰浴，向反应液加入氨水(85g)，室温搅拌30min。将反应液减压浓缩，所得浓缩物加入水(50ml)打浆。过滤，滤饼依次以水和乙醚洗涤，干燥得到标题化合物(5.8g)。MS(ESI)m/z:380.3[M-H] -.
步骤2：(2R，3R，4R，5R)-2-((双(4-甲氧基苯基)(苯基)甲氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-甲氧基四氢呋喃-3-基(2-氰基乙基)二异丙基亚磷酰胺
Figure PCTCN2019120251-appb-000019
向反应瓶中加入1-((2R，3R，4R，5R)-5-((双(4-甲氧基苯基)(苯基)甲氧基)甲基)-4-羟基-3-甲氧基四氢呋喃-2-基)嘧啶-2,4(1H，3H)-二酮(1.0g)，氮气置换后加入二氯甲烷(20ml)。依次将((双(二异丙基氨基)膦基)氧基)丙腈(0.64g)、三氟乙酸吡啶盐(0.41g)的二氯甲烷溶液(10ml)滴加入反应液中，室温反应3h。反应液浓缩， 粗品经柱层析(石油醚:乙酸乙酯＝2:1)纯化，得到标题化合物(1.2g)。MS(ESI)m/z:783.4[M+Na] +.
步骤3：((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基((2R，3R，4R，5R)-2-((双(4-甲氧基苯基)(苯基)甲氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-甲氧基四氢呋喃-3-基)(2-氰基乙基)亚磷酸酯
Figure PCTCN2019120251-appb-000020
将N-(9-((1S，3R，4S)-4-羟基-3-羟甲基-2-亚甲基环戊基)-6-氧代-6,9-二氢-1H-嘌呤-2-基)苯甲酰胺(1.263g)溶于无水乙腈(10ml)和无水N,N-二甲基甲酰胺(5ml)的混合溶剂中，加入无水乙腈(10ml×3)共沸3次。氮气保护下，依次将(2R，3R，4R，5R)-2-((双(4-甲氧基苯基)(苯基)甲氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-甲氧基四氢呋喃-3-基(2-氰基乙基)二异丙基亚磷酰胺(2.4g)的无水乙腈(10ml)溶液、0.45M1H-四唑(0.4g)的乙腈溶液(12.69ml)滴加入反应液中，室温反应2h。反应液浓缩，粗品经柱层析(二氯甲烷:甲醇＝95:5)纯化，得到标题化合物(1.41g)。
步骤4：O-(((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-2-((双(4-甲氧基苯基)(苯基)甲氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-甲氧基四氢呋喃-3-基)O-(2-氰基乙基)硫代磷酸酯
Figure PCTCN2019120251-appb-000021
反应瓶中，将((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基((2R，3R，4R，5R)-2-((双(4-甲氧基苯基)(苯基)甲氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-甲氧基四氢呋喃-3-基)(2-氰基乙基)亚磷酸酯(0.90g)溶于无水乙腈(5ml)。氮气保护下，将3H-苯并[c][1,2]二硫醇-3-酮1,1-二氧化物(0.177g)的无水乙腈(3ml)溶液滴入反应液中，室温反应1h。反应液浓缩，粗品经柱层析(二氯甲烷:甲醇＝90:10)纯化，得到标题化合物(0.7g)。
步骤5：O-(((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-(2-氰乙基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)硫代磷酸酯
Figure PCTCN2019120251-appb-000022
将O-(((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-2-((双(4-甲氧基苯基)(苯基)甲氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-甲氧基四氢呋喃-3-基)O-(2-氰基乙基)硫代磷酸酯(0.7)溶于二氯甲烷(8ml)，向反应液滴加二氯乙酸(0.129g)的二氯甲烷(1ml)溶液，室温反应4h。冰浴降温，向反应液滴加三乙胺调pH至中性。反应液浓缩，粗品经柱层析(二氯甲烷:甲醇＝90:10)纯化，得到标题化合物(0.25g)。MS(ESI)m/z:771.5[M+H] +.
步骤6：O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)硫代磷酸铵
Figure PCTCN2019120251-appb-000023
向反应瓶中依次加入O-(((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-(2-氰乙基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)硫代磷酸酯(210mg)和氨水(28％)(1ml)，室温下反应5天。冰浴下向反应液中滴加50％醋酸溶液，调节pH至8左右。反应液用乙酸乙酯萃取两次，分取水相，水相用Biotage C18 30g反相色谱柱进行分离纯化(水:乙腈＝3:1)得标题化合物(120mg)。 1H NMR(500MHz,DMSO-d 6):δ11.35(s,1H),10.52(s,1H),7.92(dd,J＝5.0Hz,8.0Hz,1H),7.71(d,J＝3.0Hz,1H),7.18(brs,4H),6.43(s,2H),5.88(d,J＝6.0Hz,1H),5.69(d,J＝8.5Hz,1H),5.37(m,1H),5.26(d,J＝5.0Hz,1H),5.13(s,1H),4.93(s,1H),4.77(m,1H),4.57(d,J＝7.0Hz,1H),4.29(s,1H),4.22(d,J＝2.5Hz,1H),4.07(d,J＝9.5Hz,1H),3.90(t,J＝6.0Hz,2H),3.65(s,2H),3.38(s,3H),2.69(s,1H),2.37(m,1H),2.04(m,1H).MS(ESI) m/z:614.5[M+H] +.
实施例2：O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1H-嘌呤-9(6H)-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)(S)-硫代磷酸钠
步骤1：(2S，3aS，6R，7aS)-3a-甲基-2-((全氟苯基)硫代)-6-(丙-1-烯-2-基)六氢苯并[d][1,3,2]氧硫杂磷杂环戊烷2-硫化物
Figure PCTCN2019120251-appb-000024
反应瓶中，依次将三乙胺双(全氟苯基)二硫代磷酸酯(126g)，(1R，4R，6S)-1-甲基-4-(丙-1-烯-2-基)-7-氧杂二环[4.1.0]庚烷(48.3g)，磷酸氢二丁酯(48.9g)加至氯仿(1200ml)中，然后将2,2-二氯乙酸(60.0g)缓慢滴加至上述溶液中，室温反应1h。将反应液浓缩至500ml后，向反应液中加入1000ml正己烷稀释，后分别10％的磷酸二氢钾溶液和饱和食盐水洗涤。将有机相浓缩至200ml，加入200ml甲醇，继续浓缩至50ml后加入200ml甲醇，再浓缩至50ml，析出固体。冰浴降温至0℃后过滤。得粗品93.48g。将粗品(93.48g)溶于225ml二氯甲烷中，后加入正庚烷374ml，粗品完全溶解。将溶液浓缩至200ml，再加入正庚烷374ml，此操作重复3次。将溶液浓缩至200ml后析出大量固体，室温搅拌2h。过滤，滤饼干燥得到标题化合物(62.58g)。 1H NMR(500MHz,CDCl 3):δ5.04(s,1H),4.87(s,1H),4.30-4.280(m,1H),2.63(s,1H),2.37-2.34(m,1H),2.07-1.85(m,4H),1.81(s,3H),1.79-1.73(m,1H),1.68(s,3H).
步骤2：1-((2R，3R，4R，5R)-5-(((叔丁基二苯基硅烷基)氧基)甲基)-3-甲氧基-4-(((2R，3aS，6R，7aS)-3a-甲基-6-(丙-1-烯-2-基)-2-磺酰基六氢苯并[d][1,3,2]氧硫杂磷杂环戊烷-2-氧基)氧基)四氢呋喃-2-基)嘧啶-2,4(1H，3H))-二酮
Figure PCTCN2019120251-appb-000025
将1-((2R，3R，4R，5R)-5-(((叔丁基二苯基硅烷基)氧基)甲基)-4-羟基-3-甲氧基四氢呋喃-2-基)嘧啶-2,4(1H，3H)-二酮(6.44g)和(2S，3aS，6R，7aS)-3a-甲基-2-((全氟苯基)硫代)-6-(丙-1-烯-2-基)六氢苯并[d][1,3,2]氧硫杂磷杂环戊烷2-硫化物(7.5g)溶于 乙腈(156ml)，冰浴降温至0℃。加入2,3,4,6,7,8,9,10-八氢嘧啶并[1,2-a]氮杂(2.57g)，0℃条件下搅拌反应5h。反应液中加入乙酸乙酯稀释，依次以纯化水和10％磷酸二氢钾溶液洗涤。有机相以无水硫酸钠干燥，过滤后将有机相浓缩得粗品。经柱层析(石油醚:乙酸乙酯＝1:1)纯化，得到标题化合物(4.53g)。 1H-NMR(500MHz,CDCl 3):δ8.72(s,1H),7.85(d,J＝8.0Hz,1H),7.69-7.64(m,4H),7.48-7.40(m,6H),6.11(d,J＝5.0Hz,1H),5.42-5.37(m,2H),5.05(s,1H),4.91(s,1H),4.59-4.55(m,1H),4.27(d,J＝4.5Hz,1H),4.04-4.01(m,1H),3.97(t,J＝5.0Hz,1H),3.89-3.86(m,1H),3.52(s,3H),2.59(s,1H),2.28(d,J＝14.0Hz,1H),2.18-2.12(m,1H),1.98-1.90(m,3H),1.78(m,4H),1.71(s,3H),1.11(s,9H).
步骤3：O-(((1R，3S，5S)-3-(2-苯甲酰基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-((叔丁基二苯基硅烷基)氧基)-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-2-(((叔丁基二苯基甲硅烷基)氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1)(2H)-基)-4-甲氧基四氢呋喃-3-基)O-氢(S)-硫代磷酸酯
Figure PCTCN2019120251-appb-000026
向反应瓶中，依次加入1-((2R，3R，4R，5R)-5-(((叔丁基二苯基硅烷基)氧基)甲基)-3-甲氧基-4-(((2R，3aS，6R，7aS)-3a-甲基-6-(丙-1-烯-2-基)-2-磺酰基六氢苯并[d][1,3,2]氧硫杂磷杂环戊烷-2-氧基)氧基)四氢呋喃-2-基)嘧啶-2,4(1H，3H))-二酮(4.5g)，N-(9-((1S，3R，4S)-4-((叔丁基二苯基硅烷基)氧基)-3-(羟甲基)-2-亚甲基环戊基)-6-氧代-6,9-二氢-1H-嘌呤吡啶-2-基)苯甲酰胺(7.51g)，后加入无水乙腈，共沸除水，重复两次。加入乙腈(52ml)，室温下缓慢滴入2,3,4,6,7,8,9,10-八氢嘧啶并[1,2-a]氮杂(2.77g)。反应3h后加入乙酸乙酯稀释，有机相依次以饱和碳酸氢钠溶液和饱和食盐水洗涤，有机层以无水硫酸钠干燥。粗品经柱层析(二氯甲烷:甲醇＝10:1)纯化，得到标题化合物(5.24g)。 1H NMR(500MHz,DMSO-d 6):δ12.53(s,1H),12.40(s,1H),11.42(d,J＝2.0Hz,1H),8.23-8.14(m,2H),7.68-7.53(m,10H),7.51-7.34(m,15H),5.88(d,J＝6.0Hz,1H),5.59(t,J＝10.0Hz,1H),5.25(d,J＝8.0Hz,1H),5.12(s,1H),4.96(m,1H),4.71(s,1H),4.67(s,1H),4.20(d,J＝9.0Hz,1H),4.12(d,J＝3.5Hz,1H),3.87-3.77(m,1H),3.76(s,2H),3.57-3.52(m,1H),3.47(t,J＝6.0Hz,1H),3.26(s,3H),2.05-1.96(m,1H),1.91(m,1H),1.03(s,9H),1.01(s,9H).
步骤4：O-(((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2- 亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-2-羟甲基-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-甲氧基四氢呋喃-3-基)(S)-硫代磷酸四丁基铵
Figure PCTCN2019120251-appb-000027
向反应瓶中，依次加入O-(((1R，3S，5S)-3-(2-苯甲酰基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-((叔丁基二苯基硅烷基)氧基)-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-2-(((叔丁基二苯基甲硅烷基)氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H))-(4-甲氧基四氢呋喃-3-基)O-氢(S)-硫代磷酸酯(5.2g)、四丁基氟化铵三水合物(3.43g)和四氢呋喃(35ml)，室温搅拌反应40h，向反应液中加入水(100ml)，然后用乙醚(50ml)萃取两次。将水相浓缩得到标题化合物7.09g，未经处理直接投入下一步。
步骤5：O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)(S)-硫代磷酸四丁基铵
Figure PCTCN2019120251-appb-000028
向反应瓶中依次加入O-(((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-2-羟甲基-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-甲氧基四氢呋喃-3-基)(S)-硫代磷酸四丁基铵(7g)和氨水(50ml)，混合物加热至50℃搅拌反应68h。冰浴下向反应液中滴加50％醋酸水溶液，调节pH至8左右。反应液用乙酸乙酯萃取两次，分取水相，水相用Biotage C18 120g反相色谱柱进行分离纯化(水:乙腈＝3:1)得到标题化合物(3.39g)。MS(ESI)m/z:614.5[M+H] +.
步骤6：O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)(S)-硫代磷酸钠
Figure PCTCN2019120251-appb-000029
向反应瓶中，依次加入O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)(S)-硫代磷酸四丁基铵(0.2g)、钠离子交换树脂(4g)及去离子水(4ml)，室温搅拌1h。过滤，水相用Biotage C18 120g反相色谱柱进行分离纯化(水:乙腈＝3:1)得到标题化合物(80mg) 1H NMR(500MHz,DMSO-d 6):δ10.69(br,1H),7.91(dd,J＝8.5,2.5Hz,1H),7.72-7.09(m,1H),6.45(s,2H),5.86(d,J＝5.7Hz,1H),5.71-5.60(m,1H),5.36-5.27(m,2H),5.11(s,1H),4.94(d,J＝3.0Hz,1H),4.76(d,J＝11.0Hz,1H),4.55(s,1H),4.28(s,1H),4.21(s,1H),4.01(q,J＝9.0Hz,1H),3.95-3.79(m,2H),3.65(s,2H),3.38(s,3H),2.70(s,1H),2.34(s,1H),2.06-1.98(m,2H).MS(ESI)m/z:614.5[M+H] +.
实施例3：O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-(2-甲氧基乙氧基)四氢呋喃-3-基)(S)-硫代磷酸钠
步骤1：1-((2R，3R，4R，5R)-5-(((叔丁基二苯基硅烷基)氧基)甲基)-3-(2-甲氧基乙氧基)-4-(((2R，3aS，6R，7aS)-3a-甲基-6-(丙-1-烯-2-基)-2-磺酰基六氢苯并[d][1,3,2]氧硫杂磷杂环戊烷-2-基)氧基)四氢呋喃-2-基)嘧啶-2,4-(1H，3H)-二酮
Figure PCTCN2019120251-appb-000030
将1-((2R，3R，4R，5R)-5-(((叔丁基二苯基硅烷基)氧基)甲基)-4-羟基-3-(2-甲氧基乙氧基)四氢呋喃-2-基)嘧啶-2,4(1H，3H)-二酮(35g)和(2S，3aS，6R，7aS)-3a-甲基-2-((全氟苯基)硫代)-6-(丙-1-烯-2-基)六氢苯并[d][1,3,2]氧硫杂磷杂环戊烷2-硫化物(37.6g)溶于乙腈(645ml)，冰浴降温。缓慢滴加2,3,4,6,7,8,9,10-八氢嘧啶并[1,2-a]氮杂卓(12.81g)至上述反应液。冰浴条件下继续反应2h。反应液依次以纯化水和10％磷酸二氢 钾溶液洗涤。有机相以无水硫酸钠干燥，过滤浓缩得粗品，经柱层析(石油醚:乙酸乙酯＝1:1)纯化，得到标题化合物(49.23g)。 1H NMR(500MHz,CDCl 3)δ8.92(s,1H),7.80(d,J＝8.2Hz,1H),7.70-7.63(m,4H),7.48-7.38(m,6H),6.12(d,J＝5.5Hz,1H),5.41-5.45(m,1H),5.38(dd,J＝8.3,1.9Hz,1H),5.05(s,1H),4.92(s,1H),4.55-4.59(m,1H),4.32-4.22(m,2H),3.98(dd,J＝11.9,2.0Hz,1H),3.90-3.79(m,3H),3.54(t,J＝4.5Hz,2H),3.30(s,3H),2.59(s,1H),2.33-2.24(m,1H),2.14(td,J＝13.4,4.2Hz,1H),2.00-1.88(m,3H),1.79(s,3H),1.71(d,J＝4.9Hz,3H),1.11(s,9H),0.91-0.82(m,1H).
步骤2：O-(((1R，3S，5S)-3-(2-苯甲酰基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-((叔丁基二苯基硅烷基)氧基)-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-2-(((叔丁基二苯基甲硅烷基)氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-(2-甲氧基乙氧基)四氢呋喃-3-基)O-氢(S)-硫代磷酸酯
Figure PCTCN2019120251-appb-000031
1-((2R，3R，4R，5R)-5-(((叔丁基二苯基硅烷基)氧基)甲基)-3-(2-甲氧基乙氧基)-4-(((2R，3aS，6R，7aS)-3a-甲基-6-(丙-1-烯-2-基)-2-磺酰基六氢苯并[d][1,3,2]氧硫杂磷杂环戊烷-2-基)氧基)四氢呋喃-2-基)嘧啶-2,4(1H，3H)-二酮(1.5g)和N-(9-((1S，3R，4S)-4-((叔丁基二苯基硅烷基)氧基)-3-羟甲基-2-亚甲基环戊基)-6-氧代-6,9-二氢-1H-嘌呤吡啶-2-基)苯甲酰胺(2.36g)溶于乙腈(20ml)，室温下加入2,3,4,6,7,8,9,10-八氢嘧啶并[1,2-a]氮杂卓(0.87g)，搅拌反应2h。反应液以乙酸乙酯稀释，依次以饱和碳酸氢钠溶液及饱和食盐水洗涤，有机相以无水硫酸钠干燥，过滤浓缩得粗品，经柱层析(二氯甲烷:甲醇＝10:1)纯化，得到标题化合物(1.98g)。 1H-NMR(500MHz,DMSO-d 6):δ12.57(s,1H),12.41(s,1H),11.41(s,1H),8.21-8.18(m,3H),7.66-7.61(m,8H),7.57-7.54(m,2H),7.47(t,J＝7.5Hz,3H),7.43-7.38(m,11H),5.86(d,J＝6.0Hz,1H),5.59(t,J＝9.5Hz,1H),5.22(d,J＝8.0Hz,1H),5.12(s,1H),4.91-4.89(m,1H),4.71(s,1H),4.67(s,1H),4.23(s,1H),4.11(s,1H),3.98(t,J＝5.5Hz,1H),3.75-3.69(m,4H),3.46-3.45(m,1H),3.27-3.26(m,2H),3.10(s,3H),2.90(s,1H),2.79(s,1H),2.04-2.00(m,1H),1.04(s,9H),1.00(s,9H).
步骤3：O-(((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基) -2-羟甲基-4-(2-甲氧基乙氧基)四氢呋喃-3-基)(S)-硫代磷酸四丁基铵
Figure PCTCN2019120251-appb-000032
向反应瓶中依次加入O-(((1R，3S，5S)-3-(2-苯甲酰基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-((叔丁基二苯基硅烷基)氧基)-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-2-(((叔丁基二苯基甲硅烷基)氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-(2-甲氧基乙氧基)四氢呋喃-3-基)O-氢(S)-硫代磷酸酯(1.80g)，四丁基氟化铵三水合物(1.38g)和四氢呋喃(20ml)，室温反应过夜。反应液用水稀释，乙醚洗涤，水相浓缩得到标题化合物2.40g，未经纯化直接投入下一步。
步骤4：O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-(2-甲氧基乙氧基)四氢呋喃-3-基)(S)-硫代磷酸四丁基铵
Figure PCTCN2019120251-appb-000033
将O-(((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-(2-甲氧基乙氧基)四氢呋喃-3-基)(S)-硫代磷酸四丁基铵(2.40g)溶于氨水(15ml)，室温反应24h。冰浴下，反应液用50％醋酸水溶液调节pH至8，乙酸乙酯洗涤。分取水相，水相用Biotage C18 120g反相色谱柱进行分离纯化(水:乙腈＝3:1)得到标题化合物(0.91g)。MS(ESI)m/z:656.7[M-H] -.
步骤5：O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-(2-甲氧基乙氧基)四氢呋喃-3-基)(S)-硫代磷酸钠
Figure PCTCN2019120251-appb-000034
将O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-(2-甲氧基乙氧基)四氢呋喃-3-基)(S)-硫代磷酸四丁基铵(0.35g)溶于去离子水(7ml)，向其中加入钠型离子交换树脂(7g)，室温搅拌1h后过滤，水相用Biotage C18 120g反相色谱柱进行分离纯化(水:乙腈＝3:1)得到标题化合物(0.17g)。 1H-NMR(500MHz,DMSO-d 6):δ10.87(brs,2H),7.91(d,J＝8.0Hz,1H),7.71(s,1H),6.46(s,2H),5.85(d,J＝5.5Hz,1H),5.67(d,J＝8.5Hz,1H),5.38-5.33(m,1H),5.27(t,J＝5.0Hz,1H),5.11(s,1H),4.94(s,1H),4.74-4.72(m,1H),4.55(s,1H),4.27(s,1H),4.20(s,1H),4.05-3.98(m,2H),3.94-3.85(m,2H),3.65(s,2H),3.61-3.57(m,1H),3.43-3.42(m,2H),3.20(s,3H),2.70(s,1H),2.39-2.34(m,1H),2.06-2.02(m,1H).MS(ESI)m/z:656.7[M-H] -.
实施例4：O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-氟-2-羟甲基-4-甲基四氢呋喃-3-基)(S)-硫代磷酸钠
步骤1：1-((2R，3R，4R，5R)-5-(((叔丁基二苯基硅烷基)氧基)甲基)-3-氟-4-羟基-3-甲基四氢呋喃-2-基)嘧啶-2,4(1H，3H)-二酮
Figure PCTCN2019120251-appb-000035
将1-((2R，3R，4R，5R)-3-氟-4-羟基-5-羟甲基-3-甲基四氢呋喃-2-基)嘧啶-2,4(1H，3H)-二酮(5g)和4-二甲氨基吡啶(0.469g)溶于吡啶(20ml)，向其中缓慢滴加叔丁基二苯基氯硅烷(6.47g)。室温搅拌12小时后，反应液用乙酸乙酯稀释，饱和氯化铵溶液洗涤。有机相以无水硫酸钠干燥，过滤浓缩得粗品，经柱层析(二氯甲烷:甲醇＝20:1)纯化，得到标题化合物(9.22g)。 1H-NMR(500MHz,DMSO-d 6):δ11.54(s,1H),7.76(d,J＝8.0Hz,1H),7.68(d,J＝7.0Hz,2H),7.63(d,J＝6.5Hz,2H),7.51-7.41(m,6H),6.08(d,J＝19.0Hz,1H),5.85(s,1H),5.11(brs,1H),4.11-4.05(m,1H),4.03-3.96(m,2H),3.94-3.89(m,1H),1.31(d,J＝22.5Hz,3H),1.01(s,9H).
步骤2：1-((2R，3R，4R，5R)-5-(((叔丁基二苯基硅烷基)氧基)甲基)-3-氟-3-甲基-4-(((2R，3aS，6R，7aS)-3a-甲基-6-(丙-1-烯-2-基)-2-硫代六氢苯并[d][1,3,2]氧硫杂磷杂环戊烷-2-基)氧基)四氢呋喃-2-基)嘧啶-2,4(1H，3H)-二酮
Figure PCTCN2019120251-appb-000036
1-((2R，3R，4R，5R)-5-(((叔丁基二苯基硅烷基)氧基)甲基)-3-氟-4-羟基-3-甲基四氢呋喃-2-基)嘧啶-2,4(1H，3H)-二酮(0.9g)和(2S，3aS，6R，7aS)-3a-甲基-2-((全氟苯基)硫代)-6-(丙-1-烯-2-基)六氢苯并[d][1,3,2]氧硫杂磷杂环戊烷2-硫化物(1.048g)溶于乙腈(16ml)，0℃下加入2,3,4,6,7,8,9,10-八氢嘧啶并[1,2-a]氮杂卓(0.357g)，搅拌反应30分钟。反应液用乙酸乙酯稀释，10％磷酸二氢钾溶液洗涤。有机相以无水硫酸钠干燥，过滤浓缩得粗品，经柱层析(石油醚:乙酸乙酯＝1:1)纯化，得到标题化合物(1.26g)。 1H-NMR(500MHz,CDCl 3):δ9.59(s,1H),7.58(d,J＝8.0Hz,1H),7.72(d,J＝7.0Hz,2H),7.66(d,J＝7.0Hz,2H),7.47-7.37(m,6H),6.32(d,J＝18.5Hz,1H),5.39-5.27(m,1H),5.08(d,J＝8.0Hz,1H),5.01(s,1H),4.87(s,1H),4.59-4.53(m,1H),4.23-4.21(m,1H),4.16-4.12(m,1H),3.84-3.82(m,1H),2.61(brs,1H),2.36-2.29(m,1H),2.16-2.08(m,1H),2.02-1.95(m,2H),1.94-1.88(m,2H),1.77(s,3H),1.73(s,3H),1.52(d,J＝22.0Hz,3H),1.11(s,9H).
步骤3：O-(((1R，3S，5S)-3-(2-苯甲酰基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-((叔丁基二苯基硅烷基)氧基)-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-2-(((叔丁基二苯基甲硅烷基)氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-氟-4-甲基四氢呋喃-3-基)O-氢(S)-硫代磷酸酯
Figure PCTCN2019120251-appb-000037
1-((2R，3R，4R，5R)-5-(((叔丁基二苯基硅烷基)氧基)甲基)-3-氟-3-甲基-4-(((2R，3aS，6R，7aS)-3a-甲基-6-(丙-1-烯-2-基)-2-硫代六氢苯并[d][1,3,2]氧硫杂磷杂环戊烷-2-基)氧基)四氢呋喃-2-基)嘧啶-2,4(1H，3H)-二酮(1.2g)和N-(9-((1S，3R，4S)-4- ((叔丁基二苯基甲硅烷基)氧基)-3-(羟甲基))-2-亚甲基环戊基)-6-氧代-6,9-二氢-1H-嘌呤-2-基)苯甲酰胺(1.997g)溶于乙腈(15ml)，室温下加入2,3,4,6,7,8,9,10-八氢嘧啶并[1,2-a]氮杂卓(0.736g)，搅拌反应30分钟。反应液用乙酸乙酯稀释，饱和碳酸氢钠溶液洗涤。有机相以无水硫酸钠干燥，过滤浓缩得粗品，经柱层析(二氯甲烷:甲醇＝10:1)纯化，得到标题化合物(1.14g)。 1H-NMR(500MHz,DMSO-d 6):δ12.56(s,1H),12.43(s,1H),11.49(s,1H),8.17(s,1H),8.14(d,J＝7.5Hz,2H),7.74-7.71(m,3H),7.69-7.61(m,6H),7.60-7.55(m,1H),7.50-7.45(m,2H),7.44-7.34(m,12H),6.02(d,J＝18.5Hz,1H),5.64-5.56(m,1H),5.06(s,1H),4.98-4.83(m,1H),4.80-4.68(m,2H),4.63(s,1H),4.14-4.05(m,1H),4.01-3.87(m,4H),3.01-2.89(m,1H),2.78(s,1H),2.08-1.99(m,1H),1.19-1.14(m,3H),1.06(s,9H),1.00(s,9H).
步骤4：O-(((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-氟-2-羟甲基-4-甲基四氢呋喃-3-基)(S)-硫代磷酸四丁基铵
Figure PCTCN2019120251-appb-000038
向反应瓶中，依次加入O-(((1R，3S，5S)-3-(2-苯甲酰基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-((叔丁基二苯基硅烷基)氧基)-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-2-(((叔丁基二苯基甲硅烷基)氧基)甲基)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-氟-4-甲基四氢呋喃-3-基)O-氢(S)-硫代磷酸酯(1g)，四丁基氟化铵三水合物(0.791g)和四氢呋喃(9ml)，室温搅拌过夜反应。反应液用水稀释，乙醚洗涤，水相浓缩得到标题化合物3g，未经纯化直接投入下一步。
步骤5：O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-氟-2-羟甲基-4-甲基四氢呋喃-3-基)(S)-硫代磷酸四丁基铵
Figure PCTCN2019120251-appb-000039
将O-(((1R，3S，5S)-3-(2-苯甲酰氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-氟-2-羟甲基-4-甲基四氢呋喃-3-基)(S)-硫代磷酸四丁基铵(3g)溶于氨水(20ml)，室温搅拌反应17h。冰浴下，反应液用50％醋酸水溶液调节pH至8，乙酸乙酯萃取。水相用Biotage C18 120g反相色谱柱进行分离纯化(水:乙腈＝3:1)得到标题化合物(0.5g)。MS(ESI)m/z:614.5[M-H] -.
步骤6：O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-氟-2-羟甲基-4-甲基四氢呋喃-3-基)(S)-硫代磷酸钠
Figure PCTCN2019120251-appb-000040
将O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-4-氟-2-羟甲基-4-甲基四氢呋喃-3-基)(S)-硫代磷酸四丁基铵(0.3g)溶于去离子水(4ml)，向其中加入钠型离子交换树脂(6g)，室温搅拌1h后过滤，水相用Biotage C18 120g反相色谱柱进行分离纯化(水:乙腈＝3:1)得到标题化合物(0.1g)。 1H-NMR(500MHz,D 2O):δ7.84(s,1H),7.82(d,J＝8Hz,1H),6.14(d,J＝19.5Hz,1H),5.85(d,J＝8Hz,1H),5.39(t,J＝8Hz,1H),5.32(s,1H),4.87(s,1H),4.68-4.62(m,1H),4.49-4.44(m,1H),4.19-1.14(m,2H),4.14-4.09(m,1H),4.02-3.98(m,1H),3.92-3.86(m,1H),2.86(s,1H),2.45-2.37(m,1H),2.30-2.23(m,1H),1.39(d,J＝23.5Hz,3H).MS(ESI)m/z:614.5[M-H] -.
实施例5：(((((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲氧基)(((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)氧基)磷酰基)硫代)甲基异丙基碳酸酯
Figure PCTCN2019120251-appb-000041
将O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环 戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)硫代磷酸铵(0.80g)溶于水(100ml)和异丙醇(26ml)的混合溶液，滴加碘甲基异丙基碳酸酯(0.845g)，室温反应24h。浓缩除去异丙醇，水相用正己烷萃取三次后，用Biotage C18 120g反相色谱柱进行分离纯化(水:乙腈＝3:1)得到标题化合物(0.536g)。 1H NMR(500MHz,DMSO-d 6)δ11.44(s,1H),10.60(s,1H),7.91(t,J＝7.5Hz,1H),7.69(d,J＝14.0Hz,1H),6.38(s,2H),5.94(dd,J＝6.5,2.0Hz,1H),5.73(d,J＝8.0Hz,1H),5.53-5.51(m,2H),5.40(dt,J＝10.0,6.5Hz,2H),5.23-5.21(m,1H),5.15(dd,J＝6.0,3.0Hz,1H),5.11-5.02(m,1H),4.83-4.80(m,1H),4.66(d,J＝2.5Hz,1H),4.30-4.21(m,3H),4.15-4.13(m,2H),3.77-3.58(m,2H),3.38(s,3H),2.85-2.84(m,1H),2.34-2.32(m,1H),2.12-2.13(m,1H),1.22(dd,J＝6.0,2.5Hz,6H).MS(ESI)m/z:730.5[M+H] +.
实施例6：(((S)-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲氧基)(((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)氧基)磷酰基)硫代)甲基异丙基碳酸酯
Figure PCTCN2019120251-appb-000042
将O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)(S)-硫代磷酸四丁基铵(1g)溶于水(100ml)和异丙醇(25ml)的混合溶液，滴加碘代甲基异丙基碳酸酯(0.856g)，室温反应24h。浓缩除去异丙醇，水相用正己烷萃取三次后用Biotage C18 120g反相色谱柱进行分离纯化(水:乙腈＝3:1)得到标题化合物(0.4g)。 1H-NMR(500MHz,CD 3OD):δ8.05(d,J＝8Hz,1H),7.79(s,1H),6.07(d,J＝6Hz,1H),5.76(d,J＝8Hz,1H),5.62-5.49(m,3H),5.36(s,1H),5.21-5.16(m,1H),4.94-4.91(m,1H),4.59-4.52(m,1H),4.51-4.44(m,2H),4.37-4.35(m,1H),4.23-4.18(m,1H),3.87-3.79(m,2H),3.52(s,3H),3.02-2.97(m,1H),2.61-2.54(m,1H),2.32-2.24(m,1H),1.88-1.78(m,1H),1.34-1.27(m,6H).MS(ESI)m/z:730.5[M+H] +.
实施例7：(((S)-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲氧基)(((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)氧基)磷酰基)硫代)甲基异丁基碳酸酯
Figure PCTCN2019120251-appb-000043
将O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)(S)-硫代磷酸四丁基铵(0.85g)溶于水(120ml)和异丙醇(30ml)的混合溶液，滴加碘代甲基异丁基碳酸酯(0.770g)，室温反应24h。浓缩除去异丙醇，水相用正己烷萃取三次后用Biotage C18 120g反相色谱柱进行分离纯化(水:乙腈＝3:1)得到标题化合物(0.15g)。 1H-NMR(500MHz,CD 3OD):δ8.05(d,J＝8.0Hz,1H),7.80(s,1H),6.07(d,J＝6.0Hz,1H),5.70(d,J＝8.0Hz,1H),5.63-5.55(m,3H),5.36(s,1H),5.20-5.19(m,1H),4.89(s,1H),4.57-4.49(m,3H),4.35(s,1H),4.20(t,J＝5.0Hz,1H),3.99(d,J＝6.5Hz,2H),3.86-3.81(m,2H),3.52(s,3H),3.00(s,1H),2.60-2.55(m,1H),2.30-2.26(m,1H),1.97-1.92(m,1H),0.95(s,3H),0.93(s,3H).MS(ESI)m/z:744.6[M+H] +.
实施例8：(((S)-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲氧基)(((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)氧基)磷酰基)硫代)甲基碳酸甲酯
Figure PCTCN2019120251-appb-000044
将O-(((1R，3S，5S)-3-(2-氨基-6-氧代-1,6-二氢-9H-嘌呤-9-基)-5-羟基-2-亚甲基环戊基)甲基)O-((2R，3R，4R，5R)-5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)-2-羟甲基-4-甲氧基四氢呋喃-3-基)(S)-硫代磷酸四丁基铵(1.0g)溶于水(112ml)和异丙醇(30ml)的混合溶液，滴加碘代甲基甲基碳酸酯(0.758g)，室温反应16h。浓缩除去异丙醇，水相用正己烷萃取三次后用Biotage C18 120g反相色谱柱进行分离纯化(水:乙腈＝3:1)，得到标题化合物(0.21g)。 1H NMR(500MHz,DMSO-d 6)δ11.44(s,1H),10.61(s,1H),7.91(s,1H),7.68(s,1H),6.41(s,2H),5.94(s,1H),5.73(s,1H),5.52(d,J＝19.5Hz,2H),5.40(s,2H),5.23(s,1H),5.11(d,J＝27.0Hz,2H),4.66(s,1H),4.26(t,J＝21.5Hz,3H),4.14(s,1H),3.77(s,2H),3.65(s,2H),3.39(s,3H),2.86(s,1H),2.51(s,2H),2.31(s,1H),2.11(s,1H).
13C NMR(126MHz,DMSO-d 6)δ163.36,157.28,154.56,153.94,151.80,151.07,149.49,140.64,136.43,116.84,111.42,103.01,85.79,84.00,81.12,75.35,70.31,68.88,67.66,60.91,58.47,55.89,55.38,51.81,39.05.
LCMS,([M+H] +)m/z:702.5.
体外抗乙肝病毒活性筛选
1.实验材料
1.1细胞
HepG2.2.15细胞
1.2化合物
受试化合物：实施例制备得到的化合物，用二甲基亚砜(DMSO)配制成20mM母液。
1.3试剂
本实验使用的试剂包括QIAamp 96 DNA Blood Kit(12)(货号Qiagen-51162)，FastStart Universal Probe Master(货号Roche-04914058001)，CellTiter-Blue检测试剂(货号Promega-G808B)。
2.实验方法
2.1化合物稀释：受试化合物起始终浓度为100μM，3倍稀释，8个浓度。
3.2.体外抗HBV活性实验和细胞毒性实验：第一天，种HepG2.2.15细胞(4×10 4细胞/孔)到96孔板，在37℃，5％CO 2培养过夜。第二天，加入含不同浓度化合物的新鲜培养液到培养孔中。第三至十天，每天吸除培养孔中旧的培养液，加入含不同浓度化合物的新鲜培养液。第十一天，收集培养孔中的上清，用于提取上清中的HBV DNA。qPCR实验检测HepG2.2.15上清中的HBV DNA含量。收集细胞上清后，加入CellTiter-Blue试剂，孵育后酶标仪检测每孔荧光值。抑制百分比：
抑制率％＝(1-样品中的HBV DNA含量/DMSO对照组中的HBV DNA含量)×100％。
细胞活力百分比：
细胞活力％＝(样品孔的荧光值-培养基对照的荧光值)/(DMSO对照组的荧光值-培养基对照的荧光值)×100％。
实验结果如表1所示。
表1.
实施例化合物 EC 50(nM) CC 50(μM)
实施例1 441 >100

Claims (15)

  1. 式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，
    Figure PCTCN2019120251-appb-100001
    R 1选自
    Figure PCTCN2019120251-appb-100002
    其中R选自以下基团：C 1-6烷基、C 2-6烯基、C 2-6炔基、C 3-6环烷基、3-6元杂环烷基、C 6-10芳基或5-10元杂芳基，所述R基团任选地被一个或多个以下基团取代：卤素、-OH、C 1-6烷基、C 1-6烷氧基、-NO 2、-CN、-NH 2、-NH(C 1-6烷基)、-N(C 1-6烷基) 2、-C(O)C 1-6烷基、-C(O)OC 1-6烷基、-CONHC 1-6烷基、-SH或-SC 1-6烷基；
    R 2及R 3分别独立地选自H、-OH、卤素、C 1-6烷基、C 1-6烷氧基、C 3-10环烷基、3-10元杂环烷基、C 6-10芳基或5-10元杂芳基，所述C 1-6烷基、C 1-6烷氧基、C 3-10环烷基、3-10元杂环烷基、C 6-10芳基或5-10元杂芳基任选地被一个或多个选自卤素、-OH、-NO 2、-NH 2或C 1-6烷氧基的取代基取代；
    R 4及R 5分别独立地选自H、-OH、-NH 2或-SH；
    n选自0、1、2、3或4；
    X选自-C(R aR b)-或-C(＝CH 2)-；
    其中，R a和R b分别独立地选自H、卤素、C 1-6烷氧基或C 1-6烷基，所述C 1-6烷氧基或C 1-6烷基任选地被一个或多个选自C 1-6烷氧基、卤素、-OH、-NH 2或-NO 2的取代基取代。
  2. 如权利要求1所述的式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，其中
    Figure PCTCN2019120251-appb-100003
    与
    Figure PCTCN2019120251-appb-100004
    结合，形成
    Figure PCTCN2019120251-appb-100005
    其中
    Figure PCTCN2019120251-appb-100006
    为一价阳离子。
  3. 如权利要求1所述式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，其中R选自以下基团：C 1-6烷基、C 2-4烯基、C 2-4炔基、C 3-6环烷基、3-6元杂环烷基、苯基或5-6元杂芳基；或者R选自C 1-4烷基；或者R选自异丙基、甲基或异丁基。
  4. 如权利要求1-3任一项所述式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，其中R 2及R 3分别独立地选自H、-OH、卤素、C 1-6烷基、C 1-6烷氧基、C 3-10环烷基、 3-10元杂环烷基、C 6-10芳基或5-10元杂芳基，所述C 1-6烷基或C 1-6烷氧基任选地被一个或多个卤素、-OH、-NO 2、-NH 2或C 1-6烷氧基的取代基取代；或者R 2及R 3分别独立地选自H、卤素、C 1-6烷基或任选地被C 1-6烷氧基取代的C 1-6烷氧基；或者R 2及R 3分别独立地选自H、F、CH 3-、CH 3O-或CH 3OCH 2CH 2O-；或者R 2为H或F，R 3为CH 3-、CH 3O-或CH 3OCH 2CH 2O-。
  5. 如权利要求1-4任一项所述式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，其中R 4及R 5分别独立地选自H或-OH；或者R 4为H，R 5为-OH。
  6. 如权利要求1-5任一项所述式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，其中n选自0、1或2；或者n为1。
  7. 如权利要求1-6任一项所述式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，其中R a和R b分别独立地选自H、卤素、C 1-3烷氧基或C 1-3烷基；或者R a和R b分别独立地选自H或卤素。
  8. 如权利要求1-7任一项所述式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，其中X为-C(＝CH 2)-。
  9. 如权利要求1-8任一项所述式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，其中所述药学上可接受的盐选自钠盐或铵盐。
  10. 如权利要求1-9任一项所述式I化合物、其互变异构体、立体异构体或其药学上可接受的盐，其中式I化合物、其互变异构体、立体异构体或其药学上可接受的盐选自式II化合物、其互变异构体、立体异构体或其药学上可接受的盐：
    Figure PCTCN2019120251-appb-100007
  11. 以下化合物、其互变异构体、立体异构体或其药学上可接受的盐：
    Figure PCTCN2019120251-appb-100008
  12. 以下化合物、其互变异构体、立体异构体或其药学上可接受的盐：
    Figure PCTCN2019120251-appb-100009
    Figure PCTCN2019120251-appb-100010
  13. 一种药物组合物，其包含权利要求1-12任一项所述式I化合物、其互变异构体、立体异构体或其药学上可接受的盐。
  14. 权利要求1-12任一项所述式I化合物、其互变异构体、立体异构体或其药学上可接受的盐、或权利要求13所述药物组合物在制备用于预防或者治疗哺乳动物HBV感染的药物中的用途；任选地，所述预防或者治疗哺乳动物HBV感染是指控制、降低或清除HBV以预防、缓解或治愈受感染的哺乳动物的肝脏疾病。
  15. 用于预防或者治疗哺乳动物HBV感染的权利要求1-12任一项所述式I化合物、其互变异构体、立体异构体或其药学上可接受的盐、或权利要求13所述药物组合物；任选地，所述预防或者治疗哺乳动物HBV感染是指控制、降低或清除HBV以预防、缓解或治愈受感染的哺乳动物的肝脏疾病。
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WO2020212301A1 (en) * 2019-04-16 2020-10-22 Roche Innovation Center Copenhagen A/S Novel process for preparing nucleotide p(v) monomers
JP2022529342A (ja) * 2019-04-16 2022-06-21 ロシュ イノベーション センター コペンハーゲン エーエス ヌクレオチドｐ（ｖ）モノマーを調製するための新規の方法
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WO2025095052A1 (ja) * 2023-11-01 2025-05-08 リードファーマ株式会社 炭素環式ヌクレオシド含有オリゴヌクレオチド

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