Nucleoside compound, pharmaceutical composition and use thereof

Abstract

The present invention discloses a nucleoside compound, a pharmaceutical composition, and uses thereof. The nucleoside is linked to a phosphate ester via a phosphorus-oxygen bond, and the nucleoside phosphoramidate compound, as described in Formulas I, Ia, and Ib, and its stereoisomers, salts, hydrates, solvates, or crystals, are described. #imgabs0# The present invention is easy to prepare and has proven efficacy.

Description

Technical Field

[0001] The present invention belongs to the field of medicine and relates to a nucleoside compound and its use as a medicine, particularly as a medicine for treating and / or preventing hepatitis B. The present invention also relates to a composition of these nucleoside compounds with other antiviral agents and its use in treating and / or preventing hepatitis B virus (HBV) infection. Background Art

[0002] Hepatitis B virus (HBV) belongs to the Hepatoviridae family. It can cause acute and / or progressive chronic illness. Acute HBV in adults is generally self-resolving through immune mechanisms. However, chronic hepatitis B (CHB) has become a significant global healthcare challenge and a major cause of chronic liver disease, cirrhosis, and hepatocellular carcinoma (HCC). The World Health Organization estimates that 2 billion people worldwide are chronically infected with HBV, with over 350 million chronically infected individuals. Nearly 600,000 people die annually from liver failure, cirrhosis, and hepatocellular carcinoma caused by HBV infection. Currently, the mainstay of treatment for chronic hepatitis B (CHB) is antiviral therapy. Interferon alpha (IFN-a), pegylated IFN-a, and nucleoside (acid) analogs (lamivudine, adefovir dipivoxil, entecavir, telbivudine, and tenofovir) are approved by the US Food and Drug Administration (FDA) for clinical treatment. Interferon was the first FDA-approved anti-HBV drug. It primarily eliminates the virus through direct antiviral effects and by inducing an immune response. However, its application is limited by its low response rate, numerous side effects, high cost, and limited therapeutic targets. Nucleoside (acid) drugs for HBV resistance share a common characteristic: they specifically target viral DNA polymerase, effectively inhibiting viral replication and are better tolerated by patients than interferon. L-FMAU has a low bioavailability of approximately 20%, and its metabolic half-life is short. Both a short half-life and low bioavailability can affect the clinical therapeutic efficacy of L-FMAU. A short half-life shortens the dosing interval, while low bioavailability increases the patient's dosage, increasing the metabolic burden on the patient and leading to significant side effects from long-term use. The present invention relates to a series of D-amino acid ester prodrugs of L-FMAU that not only improve the bioavailability of the parent drug but also significantly enhance liver targeting, thereby reducing the drug's toxic side effects. Summary of the Invention

[0003] The present invention relates to a novel class of nucleoside compounds and their use in preparing medicaments for treating and preventing HBV infection. The novel nucleoside compounds of the present invention have excellent stem cell targeting, good pharmacokinetic properties, good solubility, low toxicity, and good liver microsomal stability, and have promising application prospects in the treatment of HBV.

[0004] The present invention relates to a nucleoside compound characterized in that: a nucleoside is connected to a phosphate ester via a phosphorus-oxygen bond, and the structure is as described in general formulas I, Ia and Ib, a nucleoside phosphoramidate compound, a stereoisomer, a salt, a hydrate, a solvate or a crystal thereof,

[0005]

[0006] in:

[0007] R is selected from C1-10 alkyl, halogenated C1-10 alkyl, alkoxy, heteroalkyl, heterocycloalkyl; deuterated C1-10 alkyl, deuterated halogenated C1-10 alkyl; deuterated alkoxy, heteroalkyl, heterocycloalkyl; alkyl includes straight-chain, branched and cyclic alkyl; Ar is selected from phenyl, biphenyl, naphthyl, heteroaryl; substituted phenyl, biphenyl, naphthyl, heteroaryl; deuterated phenyl, biphenyl, naphthyl, heteroaryl; deuterated substituted phenyl, biphenyl, naphthyl, heteroaryl; preferably, Ar is selected from phenyl, biphenyl, amino acid ester substituted phenyl; deuterated phenyl, biphenyl, amino acid ester substituted phenyl.

[0008] The nucleoside phosphoramidate compound, the structure of which is as described in general formula II, IIa, IIb, III, IIIa, IIIb, IV, IVa and IVb, or its stereoisomers, salts, hydrates, solvates or crystals,

[0009]

[0010]

[0011] (1) R and R1 are selected from C1-10 alkyl, halogenated C1-10 alkyl, alkoxy, heteroalkyl, heterocycloalkyl; deuterated C1-10 alkyl, deuterated halogenated C1-10 alkyl; deuterated alkoxy, heteroalkyl, heterocycloalkyl; alkyl includes straight-chain, branched and cyclic alkyl; selected from C1-10 alkyl, halogenated C1-10 alkyl, alkoxy, heteroalkyl, heterocycloalkyl; deuterated C1-10 alkyl, deuterated halogenated C1-10 alkyl; deuterated alkoxy, heteroalkyl, heterocycloalkyl; alkyl includes straight-chain, branched and cyclic alkyl;

[0012] (2) R2 is selected from amino, C1-6 amine, C1-10 amide or alkoxyamide; including linear, branched or cyclic amine, amide or alkoxyamide; or substituted linear, branched or cyclic amine, amide or alkoxyamide;

[0013] The nucleoside phosphoramidate compound is characterized by:

[0014] Wherein R and R1 are independently H or have the following groups:

[0015]

[0016] Wherein R2 is amino or has the following groups:

[0017]

[0018] The nucleoside phosphoramidate compound having antiviral effects is characterized in that the compound is selected from the following structures, but is not limited to these compounds:

[0019]

[0020]

[0021]

[0022] In another aspect, the present invention provides use of the compound or the pharmaceutical composition in preparing a medicament, wherein the medicament is used to prevent, treat, cure or alleviate viral infectious diseases in patients.

[0023] In some embodiments, the pharmaceutical composition is an antiviral pharmaceutical composition, which optionally further comprises one or more therapeutic agents.

[0024] A use of the pharmaceutical composition in preparing a preparation having anti-HBV infection effect.

[0025] The invention is easy to prepare and has definite effect. Detailed Description of the Invention

[0027] Unless expressly provided otherwise, the following definitions apply:

[0028] As used herein, the term "halogen" or halo refers to fluorine, bromine, chlorine, or iodine, particularly when attached to an alkyl group, and also includes bromine or iodine when on an aryl or heteroaryl group.

[0029] As used herein, unless otherwise indicated, the term "heteroatom" refers to a nitrogen (N), oxygen (O), or sulfur (S) atom.

[0030] As used herein, the term "alkyl" refers to a fully saturated branched or unbranched hydrocarbon moiety having up to 10 carbon atoms. Unless otherwise provided, an alkyl group refers to a hydrocarbon moiety having 1 to 6 carbon atoms. Representative examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl, etc. Substituted alkyl groups are alkyl groups containing one or more substituents instead of hydrogen, such as 1,2 or 3 substituents, up to the number of hydrogen present on the unsubstituted alkyl group. If not otherwise stated, suitable substituents for alkyl may be selected from halogen, CN, oxo, hydroxy, C1-4 alkoxy, substituted or unsubstituted C3-6 cycloalkyl, substituted or unsubstituted phenyl, amino, (C1-4 alkyl)amino, di(C1-4 alkyl)amino, C1-4 alkylthio, C1-4 alkylsulfonyl, -C(=O)-C1-4 alkyl, COOH, COO(C1-4 alkyl), -O(C=O)-C1-4 alkyl, -NHC(=O)C1-4 alkyl and -NHC(=O)OC1-4 alkyl; wherein, for substituted cycloalkyl or phenyl, the substituents are up to three groups selected from Me, Et, -OMe, -OEt, CF3, halogen, CN, OH and NH2.

[0031] As used herein, the term "haloalkyl" refers to an alkyl group, as defined herein, substituted with one or more halogen groups. A haloalkyl group may be a monohaloalkyl, a dihaloalkyl, a trihaloalkyl, or a polyhaloalkyl group, including a perhaloalkyl group. A monohaloalkyl group may have one chlorine or one fluorine group within the alkyl group. Chlorine and fluorine are typically present as substituents on alkyl or cycloalkyl groups; fluorine, chlorine, and bromine are typically present on aryl or heteroaryl groups. Dihaloalkyl and polyhaloalkyl groups may have two or more identical halogen atoms or a combination of different halogen groups on the alkyl group. Typically, a polyhaloalkyl group contains up to 12, 10, 8, 6, 4, 3, or 2 halogen groups. Non-limiting examples of haloalkyl groups include fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, heptafluoropropyl, difluorochloromethyl, dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl, and dichloropropyl. A perhaloalkyl group refers to an alkyl group in which all hydrogen atoms are replaced by halogen atoms, such as trifluoromethyl.

[0032] As used herein, the term "alkoxy" refers to an alkyl-O- group, wherein alkyl is as defined above. Representative examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, tert-butoxy, pentoxy, hexoxy, and the like. Typically, an alkoxy group has 1 to 6 carbon atoms, more typically 1 to 4 carbon atoms.

[0033] As used herein, the term "haloalkoxy" refers to a haloalkyl-O- group, wherein haloalkyl is as defined above. Representative examples of haloalkoxy include, but are not limited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy, trichloromethoxy, 2-chloroethoxy, 2,2,2-trifluoroethoxy, 1,1,1,3,3,3-hexafluoro-2-propoxy, and the like. Typically, a haloalkyl group has 1-4 carbon atoms.

[0034] As used herein, the term "cycloalkyl" refers to a saturated or unsaturated non-aromatic monocyclic, bicyclic, tricyclic or spirocyclic hydrocarbon radical of 3-12 carbon atoms: a cycloalkyl radical may be unsaturated and may be fused to another ring which may be saturated, unsaturated or aromatic, provided that the ring atom of the cycloalkyl radical attached to the subject molecular formula is not an aromatic ring carbon. Unless otherwise specified, a cycloalkyl radical refers to a cyclic hydrocarbon radical having 3 to 9 ring carbon atoms or 3 to 7 ring carbon atoms. Preferably, a cycloalkyl radical is a saturated monocyclic ring having 3 to 7 ring atoms, such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl, unless otherwise specified.

[0035] Substituted cycloalkyl is a cycloalkyl that is substituted by one, two, three, or more than three substituents, up to the number of hydrogens on the unsubstituted group. Typically, unless otherwise indicated, a substituted cycloalkyl will have 1-4 substituents. Unless otherwise indicated, suitable substituents are independently selected from halogen, hydroxy, thiol, cyano, nitro, oxo, C1-C4 alkylimino, C1-C4-alkoxyimino, hydroxyimino, C1-C4 alkyl, C2-C4-alkenyl, C2-C4-alkynyl, C1-C4-alkoxy, C1-C4-thioalkyl, C2-C4-alkenyloxy, C2-C4-alkynyloxy, C1-C4-alkylcarbonyl, carboxyl, C1-C4-alkoxycarbonyl, amino, C1-C4-alkylamino, di-C1-C4-alkylamino, -C4-alkylamino, C1-C4-alkylaminocarbonyl, di-C1-C4-alkylaminocarbonyl, C1-C4-alkylcarbonylamino, C1-C4-alkylcarbonyl(C1-C4-alkyl)amino, C1-C4-alkylsulfonyl, C1-C4-alkylsulfamoyl and C1-C4-alkylaminosulfonyl, wherein each of the above hydrocarbon groups (e.g., alkyl, alkenyl, alkynyl, alkoxy residues) may be further substituted with one or more groups independently selected at each occurrence from the list of "alkyl" substituents herein. Preferred substituents for cycloalkyl include C1-C4-alkyl and the substituents listed above as suitable substituents for alkyl.

[0036] As used herein, the term "aryl" refers to an aromatic hydrocarbon group having 6-14 carbon atoms in the ring portion. Typically, an aryl group is a monocyclic, bicyclic, or tricyclic aromatic group having 6-14 carbon atoms, typically 6-10 carbon atoms, such as a phenyl or naphthyl group. Additionally, the term "aryl" as used herein refers to an aromatic substituent, which may be a single aromatic ring or a plurality of aromatic rings fused together. Non-limiting examples include phenyl, naphthyl, and 1,2,3,4-tetrahydronaphthyl, provided that the tetrahydronaphthyl group is connected to the formula via a carbon of the aromatic ring of the tetrahydronaphthyl group. Unless otherwise indicated, a preferred aryl group is phenyl. Substituted aryl is aryl substituted with 1-5 (e.g., one, two, or three) substituents independently selected from hydroxy, thiol, cyano, nitro, C1-C4 alkyl, C2-C4-alkenyl, C2-C4-alkynyl, C1-C4-alkoxy, C1-C4-thioalkyl, C2-C4-alkenyloxy, C2-C4-alkynyloxy, halogen, C1-C4-alkylcarbonyl, carboxyl, C1-C4-alkoxycarbonyl, amino, C1-C4-alkylamino, di-C1-C4-alkylamino , C1-C4-alkylaminocarbonyl, di-C1-C4-alkylaminocarbonyl, C1-C4-alkylcarbonylamino, C1-C4-alkylcarbonyl(C1-C4-alkyl)amino, C1-C4-alkylsulfonyl, sulfamoyl, C1-C4-alkylsulfamoyl and C1-C4-alkylaminosulfonyl, wherein each of the above hydrocarbon groups (e.g., alkyl, alkenyl, alkynyl, alkoxy residues) may be further substituted by one or more groups independently selected at each occurrence from the group listed above as suitable substituents for alkyl. Preferred substituents for substituted aryl are C1-4 alkyl, as well as those groups listed above as suitable substituents for alkyl, excluding divalent groups such as oxo.

[0037] Similarly, each cycloalkyl portion of other groups such as "aryloxy" and "aryloxyalkyl" shall have the same meaning as described in the above definition of "aryl".

[0038] As used herein, the term "heterocyclyl" refers to a saturated or partially unsaturated but non-aromatic heterocyclic group, and may be monocyclic or polycyclic (in the case of polycyclics, particularly bicyclic, tricyclic or spirocyclic); and has 3 to 14, more commonly 4 to 10, most preferably 5 or 6 ring atoms; wherein one or more, preferably one to four, especially one or two ring atoms are heteroatoms independently selected from O, S and N (the remaining ring atoms are therefore carbon). Even if described as, for example, a C5-6 atom ring, the heterocycle contains at least one heteroatom as a ring atom, the other ring atoms are carbon, and have the number of ring atoms, for example, 5-6 in this example. Preferably, the heterocyclic group has one or two such heteroatoms as ring atoms, and preferably, the heteroatoms are not directly connected to each other. Unless otherwise stated, the bonded ring (i.e., the ring connected to the target formula) preferably has 4-12, particularly 5-7 ring atoms. The heterocyclic group may be fused to an aromatic ring, provided that the atoms of the heterocyclic group connected to the target chemical formula are not aromatic. The heterocyclic group can be connected to the target formula through a heteroatom (usually nitrogen) or a carbon atom of the heterocyclic group. The heterocyclic group can include fused rings or bridged rings and spirocycles, and as long as one ring of the polycyclic heterocyclic group contains a heteroatom as a ring atom. The example of heterocycle includes tetrahydrofuran (THF), dihydrofuran, 1,4-dioxane, morpholine, 1,4-dithiane, piperazine, piperidine, 1,3-dioxolane, imidazolidine, imidazoline, pyrroline, pyrrolidine, tetrahydropyran, dihydropyran, oxathiolane, dithiolane, 1,3-dioxane, 1,3-dithiane, oxathiolane, thiomorpholine etc.

[0039] Substituted heterocyclic groups are heterocyclic groups independently substituted with 1 to 5 (eg, one, two, or three) substituents selected from the substituents for the cycloalkyl groups described above.

[0040] Similarly, each heterocyclyl portion of other groups such as "heterocyclyloxy" and "heterocyclyloxyalkyl" shall have the same meaning as described in the above definition of "heterocyclyl".

[0041] As used herein, the term "heteroaryl" refers to a 5-14 membered monocyclic- or bicyclic- or tricyclic-aromatic ring system having 1 to 8 heteroatoms as ring members, the remaining ring atoms being carbon, and the heteroatoms being selected from N, O and S. Typically, the heteroaryl group is a 5-10 membered ring system, in particular a 5-6 membered monocyclic or 8-10 membered bicyclic group. Typical heteroaryl groups include 2- or 3-thienyl, 2- or 3-furyl, 2- or 3-pyrrolyl, 2-, 4- or 5-imidazolyl, 1-, 3-, 4- or 5-pyrazolyl, 2-, 4- or 5-thiazolyl, 3-, 4- or 5-isothiazolyl, 2-, 4- or 5-oxazolyl, 3-, 4- or 5-isoxazolyl, 3- or 5-1,2,4-triazolyl, 4- or 5-1,2,3-triazolyl, 1- or 2-tetrazolyl, 2-, 3- or 4-pyridinyl, 3- or 4-pyridazinyl, 3-, 4- or 5-pyrazinyl, 2-pyrazinyl and 2-, 4- or 5-pyrimidinyl.

[0042] The term "heteroaryl" also refers to a group in which a heteroaromatic ring is fused to one or more aryl, cycloalkyl, or heterocyclyl rings. Non-limiting examples include 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl.

[0043] Substituted heteroaryl groups are heteroaryl groups which contain one or more substituents, typically one or two substituents, selected from the substituents listed above as suitable for aryl groups.

[0044] Similarly, each heteroaryl portion of other groups such as "heteroaryloxy" and "heteroaryloxyalkyl" shall have the same meaning as described in the above definition of "heteroaryl".

[0045] In many cases, the compounds of the present invention are capable of forming acid and / or base salts due to the presence of amino and / or carboxyl groups or similar groups. As used herein, the term "salt" or "salts" refers to acid addition salts or base addition salts of the compounds of the present invention. "Salts" specifically include "pharmaceutically acceptable salts." The term "pharmaceutically acceptable salts" refers to salts that retain the biological effectiveness and properties of the compounds of the present invention and are generally not biologically or otherwise undesirable.

[0046] Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids.

[0047] Any formula given herein is also intended to represent the unlabeled form of the compound (i.e., all atoms in the compound are present at their natural isotopic abundances and are non-isotopically enriched) as well as isotopically enriched or labeled forms. An isotopically enriched or labeled compound has a structure described by the general formula given herein, except that at least one atom of the compound is replaced by an atom having an atomic mass or mass number different from that of the naturally occurring atomic mass or atomic mass distribution. Examples of isotopes that can be incorporated into the enriched or labeled compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as 2H, 3H, 11C, 13C, 14C, 15N, 18F, 31P, 32P, 35S, 36Cl, and 125I, respectively. The present invention includes various isotopically labeled compounds as defined herein, such as those compounds in which radioactive isotopes such as 3H and 14C, or in which the presence of non-radioactive isotopes such as 2H and 13C is significantly higher than the natural abundance of these isotopes. These isotopically labeled compounds can be used for metabolic studies (e.g., with 14C), reaction kinetic studies (e.g., with 2H or 3H), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug or substrate tissue distribution assays, or for radiotherapy of patients. In particular, 18F-labeled compounds may be particularly ideal for PET or SPECT studies. Isotopically labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described in the accompanying examples using appropriate isotopically labeled reagents instead of otherwise used unlabeled reagents. In addition, substitution with heavier isotopes, particularly deuterium (i.e., 2H or D), can provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements or improved therapeutic index. The concentration of such heavier isotopes, particularly deuterium, can be defined by an isotopic enrichment factor. As used herein, the term "isotopic enrichment factor" means the ratio between the isotopic abundance of a specified isotope and its natural abundance. If a substituent in a compound of the invention is represented as deuterium, such compound has a deuterium isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation). Pharmaceutically acceptable solvates according to the present invention include those wherein the solvent of crystallization may be isotopically substituted, such as D2O, d6-acetone, d6-DMSO as well as solvates with non-enriched solvents.

[0048] As used herein, the term "inhibit" refers to reducing or suppressing a given condition, symptom or disorder or disease, or significantly reducing the baseline activity of a biological activity or process.

[0049] As used herein, the terms "treat," "treat," or "manage" of any disease or disorder refer, in one embodiment, to ameliorating the disease or disorder (i.e., slowing or arresting or reducing the development of the disease or at least one clinical symptom thereof). In another embodiment, "treat," "treat," or "manage" refers to alleviating or improving at least one physical parameter, including those that may not be discernible by the patient. In another embodiment, "treat," "treat," or "manage" refers to modulating the disease or disorder physically (e.g., stabilizing an identifiable symptom), physiologically (e.g., stabilizing a physical parameter), or both. In another embodiment, "treat," "treat," or "manage" refers to preventing or delaying the development or progression of a disease or disorder.

[0050] All methods described herein can be performed in any suitable order, unless otherwise noted herein or otherwise clearly contradicted by context. The use of any and all examples or exemplary language (e.g., "such as") provided herein is intended only to better illustrate the present invention, rather than to limit the scope of the present invention otherwise claimed.

[0051] Any asymmetric atoms (e.g., carbon, etc.) of the compounds of the present invention may exist in racemic or enantiomerically enriched forms, such as (R)-, (S)-, or (R, S)-configurations. In certain embodiments, each asymmetric atom has an (R)- or (S)-configuration of up to 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% or at least 99% enantiomeric excess; that is, for optically active compounds, it is generally preferred to use one enantiomer to the substantial exclusion of the other, and thus an enantiomeric purity of at least 95% is generally preferred. Substituents on atoms with unsaturated double bonds may exist in cis-(Z)- or trans-(E)-forms, if possible. Thus, as used herein, the compounds of the invention may be in the form of one of the possible isomers, rotamers, atropisomers, tautomers, or mixtures thereof, such as substantially pure geometric (cis or trans) isomers, diastereomers, optical isomers (enantiomers), racemates, or mixtures thereof. As used herein, "substantially pure" or "substantially free of other isomers" means that the product contains less than 5% by weight, and preferably less than 2% by weight, of other isomers relative to the preferred isomer. The resulting isomeric mixture can be separated into pure or substantially pure geometric or optical isomers, diastereomers, racemates, generally based on the physicochemical differences of the components, such as by chromatography and / or fractional crystallization.

[0052] The racemate of the final product or intermediate can be split into optical antipodes by known methods, for example, by separating its diastereomeric salts obtained with optically active acids or bases, and releasing optically active acidic or basic compounds. Specifically, the basic moiety can thus be used to split the compounds of the present invention into their optical antipodes, for example, by fractional crystallization of salts formed with optically active acids such as tartaric acid, dibenzoyltartaric acid, diacetyltartaric acid, two-O, O'-toluoyltartaric acid, mandelic acid, malic acid or camphor-10-sulfonic acid. The racemic product can also be split by chiral chromatography, for example, using high pressure liquid chromatography (HPLC) of a chiral stationary phase.

[0053] In addition, the compounds of the present invention, including their salts, can also be obtained in the form of their hydrates, or include other solvents used for their crystallization. The compounds of the present invention can inherently or by design form solvates with pharmaceutically acceptable solvents (including water); therefore, the present invention is intended to include both solvated and non-solvated forms. The term "solvate" refers to a molecular complex of a compound of the present invention (including its pharmaceutically acceptable salt) and one or more solvent molecules. Such solvent molecules are those commonly used in the pharmaceutical field and are known to be harmless to the recipient, such as water, ethanol, etc. The term "hydrate" refers to a complex in which the solvent molecule is water.

[0054] On the other hand, the present invention provides pharmaceutical compositions, which include the compounds of this invention or its pharmaceutically acceptable salt and at least one pharmaceutically acceptable carrier. In some embodiments, the composition includes at least two pharmaceutically acceptable excipients or carriers. Pharmaceutically acceptable carriers and other excipients are well known to those skilled in the art, and can be selected from, for example, carriers and excipients used in approved (registered) formulations of therapeutic agents administered by similar routes of administration. Pharmaceutical compositions can be formulated for specific routes of administration, such as oral administration, parenteral administration, and rectal administration. In addition, the pharmaceutical compositions of the present invention can be made in solid form (including but not limited to capsules, tablets, pills, granules, powders, or suppositories) or in liquid form (including but not limited to solutions, suspensions, or emulsions). Pharmaceutical compositions can undergo conventional drug operations, such as sterilization and / or can include conventional inert diluents, lubricants, or buffers, and adjuvants such as preservatives, stabilizers, wetting agents, emulsifiers, and buffers. In one embodiment, the compounds of this invention are formulated for oral delivery. Typically, these pharmaceutical compositions are tablets or gelatin capsules comprising the active ingredient (at least one compound of formula (I)) and one or more excipients selected from:

[0055] a) diluents, for example lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine; b) lubricants, for example silicon dioxide, talc, stearic acid, its magnesium or calcium salts and / or polyethylene glycol; c) binders, for tablets, for example magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidone; d) disintegrants, for example starch, agar, alginic acid or its sodium salt or effervescent mixtures, if desired; and / or e) absorbents, colorants, flavorings and sweeteners.

[0056] Tablets may be film coated or enteric coated according to methods known in the art.

[0057] Suitable compositions for oral administration include an effective amount of a compound of the present invention in the form of tablets, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use are prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more agents selected from sweeteners, flavorings, colorants, and preservatives to provide pharmaceutically elegant and palatable products. Tablets may contain the active ingredient mixed with non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets. These excipients are, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrants such as corn starch or alginic acid; binders such as starch, gelatin, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. The tablets are uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained effect over a longer period of time. For example, a time-delaying material such as glyceryl monostearate or glyceryl distearate can be used. Formulations for oral administration can be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oily base, such as peanut oil, liquid paraffin, or olive oil. Certain injectable compositions are aqueous isotonic solutions or suspensions, and suppositories are advantageously prepared from fat emulsions or suspensions. The compositions can be sterilized and / or contain adjuvants such as preservatives, stabilizers, wetting agents or emulsifiers, solution promoters, salts for regulating osmotic pressure, and / or buffers. In addition, they may also contain other therapeutically valuable substances. The compositions are prepared according to conventional mixing, granulation, or coating methods, respectively, and contain approximately 0.1-75% or approximately 1-50% of the active ingredient.

[0058] The compounds of the invention and intermediates can also be converted into each other according to methods generally known to those skilled in the art.

[0059] The present invention will be further described below with reference to the embodiments. DETAILED DESCRIPTION

[0060] The present invention will be further described in detail below with reference to the following examples, but the present invention is not limited to these examples. The reagents and raw materials used in the present invention were all commercially available.

[0061] Example 1

[0062]

[0063] Synthesis of compounds 2 and 3

[0064] To a solution of phenyl dichlorophosphate (4.20 mL, 27.4 mmol) in isopropyl acetate (36 mL) was added dropwise a solution of D-alanine isopropyl ester 1 (3.6 g, 27.4 mmol) and triethylamine (4.20 mL, 30.2 mmol) in isopropyl acetate (20 mL) at -10°C, while maintaining the temperature below -5°C. After stirring for 4 hours, a solution of 2,3,4,5,6-pentafluorophenol (5.56 g, 30.2 mmol) and triethylamine (4.4 mL, 31.6 mmol) in isopropyl acetate (10 mL) was added dropwise, while maintaining the temperature below -5°C. After the addition was complete, the temperature was raised to 10°C. After stirring for 18 hours, the reaction mixture was filtered and washed with isopropyl acetate. The mixture was concentrated to approximately 50 mL under vacuum distillation, and then heptane (80 mL) was added while maintaining a constant volume and an internal temperature of 40°C. The mixture was cooled to 20°C and stirred for 20 hours. The solid was filtered, washed with isopropanol / water (1:1), and dried under vacuum overnight to give (R,R)-2 (9.6 g; 76% yield; >99% dr). The mother liquor was separated on a silica gel column (50% ethyl acetate / hexane) to give (R,S)-3 (1.2 g; 9.5%). (R,R)-2: 1 H NMR(400MHz, CDCl3)δ(ppm):7.39–7.35(m,2H),7.34–7.21(m,3H),5.18–4.97(m,1H),4.28–4.0 9(m,1H),3.97–3.92(m,1H),1.46(d,J=7.0Hz,3H),1.26(d,J=5.9Hz,3H),1.25(d,J=5.9Hz,3H). 31 P NMR (162MHz, CDCl3) δ-1.83. (R, S)-3: 1H NMR(400MHz, CDCl3)δ(ppm):7.37–7.33(m,2H),7.32–7.19(m,3H),5.20–4.95(m,1H),4.18–4.0 1(m,1H),3.96–3.90(m,1H),1.48(d,J=7.0Hz,3H),1.25(d,J=5.9Hz,3H),1.24(d,J=5.9Hz,3H). 31 P NMR (162MHz,CDCl3)δ-1.51.

[0065] Example 2

[0066]

[0067] In 30 minutes, thionyl chloride (8.0g, 4.92mL, 67.3mmol) was dropwise added to the suspension of D-Ala (5.0g, 56.1mmol) in isobutanol (50mL). The mixture was heated to gentle reflux for 5 hours, concentrated under reduced pressure (water bath was set to 60 ℃). After grinding with ether (20 milliliters), the gained thick jelly solidified. White powder and ether (20mL) were ground again, collected by filtration under argon gas flow, then dried under high vacuum for 18 hours to obtain (D)-2-aminopropionic acid isobutyl ester hydrochloride 5 (9.1g, 89.3%).

[0068] A solution of phenyl dichlorophosphate (6.3 mL, 42.33 mmol) in dichloromethane (80 mL) was cooled to 0 ° C, followed by the addition of (D)-2-aminopropionic acid isobutyl ester hydrochloride (7.69 g, 42.33 mmol). The mixture was further cooled to -78 ° C and a solution of triethylamine (12.4 mL) in dichloromethane (80 mL) was added dropwise over 1 hour. The mixture was stirred for another 1.5 hours at this temperature. The reaction mixture was filtered through a sand-core glass funnel, and the filtrate was concentrated under reduced pressure. The residue was ground with methyl tert-butyl ether (60 mL), filtered, and rinsed with methyl tert-butyl ether (2 x 60 mL). The combined filtrate was concentrated under reduced pressure to give compound 6 (13 g, 100%), which was used for the coupling reaction without further purification.

[0069]

[0070] Compounds 7, 8, 9, and 10 were synthesized using the same method.

[0071] Example 3

[0072]

[0073] Synthesis of compound 12

[0074] Sodium carbonate (424 mg, 4.0 mmol) and D-tyrosine methyl ester (760 mg, 4.0 mmol) were mixed in acetone / water (3:1 ratio, 20 mL). Acetic anhydride (416 μL, 4.4 mmol) was then added at room temperature, and the mixture was stirred for 1 h. The mixture was concentrated under reduced pressure, and the residue was separated on a silica gel column (linear gradient elution, 0-15% MeOH / CHCl₃) to afford 12 (783 mg, 82%) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ9.23 (s, 1H), 8.27 (d, J = 7.8Hz, 1H), 7.00-6.95 (m, 2H), 6.69-6.60 (m, 2H), 4.34(m,1H),3.58(s,3H),2.87(dd,J=13.8,5.8Hz,1H),2.75(dd,J=13.8,9.0Hz,1H),1.78(s,3H).

[0075]

[0076] Compounds 13, 14, and 15 were synthesized using the same method.

[0077] Example 4

[0078]

[0079] N-((Benzyloxy)carbonyl)-D-tyrosine 16 (1.0 g, 3.18 mmol) was dissolved in isopropanol (10 mL), and thionyl chloride (462 μL, 6.36 mmol) was added. The reaction mixture was stirred at room temperature for 72 h. The mixture was concentrated under reduced pressure, and the residue was separated on a silica gel column (linear gradient elution, 0-15% MeOH / CHCl3) to afford the product 17 (900 mg, 80%) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ9.28 (s, 1H), 7.70 (d, J = 8.0Hz, 1H), 7.41-7.19 (m, 5H), 7.08-6.98 (m, 2H), 6.72-6.63 (m, 2H), 5.08-4.93 (m, 2H), 4.93-4.80(m,1H),4.14(m,1H),2.89(dd,J=13.8,5.8Hz,1H),2.78(dd,J=13.8,9.5Hz,1H),1.16(d,J=6.2Hz,3H),1.08(d,J=6.2Hz,3H).

[0080]

[0081] Compounds 18, 19, and 20 were synthesized by a similar method.

[0082] Example 5

[0083]

[0084] Synthesis of compound 21

[0085] Under argon, compound N-((benzyloxy)carbonyl)-D-tyrosine 16 (1.0 g, 3.18 mmol), isoamyl alcohol (362 μL, 3.34 mmol), triphenylphosphine (876 mg, 3.34 mmol), and tetrahydrofuran (16 mL) were added to a reaction flask. The mixture was cooled to 0°C and DIAD (656 μL, 3.34 mmol) was added dropwise. After the addition, the mixture was warmed to room temperature and stirred for 30 minutes until the reaction was complete. The mixture was concentrated under reduced pressure, and the residue was separated on a silica gel column (linear gradient elution, 0-50% (ethyl acetate:methanol, 9:1 ratio) / hexane) to obtain compound 21 (1.2 g, 83%) as a white solid. 1 H NMR(400MHz, DMSO-d6)δ9.22(s,1H),7.76(d,J=8.0Hz,1H),7.38-7.23(m,5H),7.06-6.96(m,2H),6.70-6.60(m,2H),5.04-4.92(m,2H),4.16(m 1H),4.09-3.98(m,2H),2.88(dd,J=13.8,5.7Hz,1H),2.75(dd,J=13.8,9.7Hz,1H), 1.63-1.46(m,1H),1.42-1.34(m,2H),0.85(d,J=4.0Hz,3H),0.84(d,J=4.0Hz,3H).

[0086]

[0087] Compounds 22, 23, and 24 were synthesized using the same method.

[0088] Example 6

[0089]

[0090] Synthesis of compound 25

[0091] Phosphorus oxychloride (0.2 mL, 2.15 mmol) was added to anhydrous diethyl ether (10 mL). A solution of compound 19 (770 mg, 2.15 mmol) and anhydrous triethylamine (0.3 mL, 2.15 mmol) in anhydrous diethyl ether (5 mL) was added dropwise at –80°C. The reaction mixture was stirred vigorously at –80°C for 1 hour and then allowed to warm to room temperature over 16 hours. Triethylamine hydrochloride was filtered out, and the filtrate was evaporated to dryness under reduced pressure to obtain a clear, liquid crude product. Dichloromethane (8 mL) was added to the crude product, and the solution was cooled to 0°C. Then, isopropyl (D)-2-aminopropionate (282 mg, 2.15 mmol) was added. The mixture was further cooled to –78°C, and a solution of triethylamine (0.3 mL, 2.15 mmol) in dichloromethane (8 mL) was added dropwise over 30 minutes. The mixture was stirred at this temperature for an additional 1.5 hours. The reaction mixture was filtered through a fritted glass funnel, and the filtrate was concentrated under reduced pressure. The residue was triturated with methyl tert-butyl ether (5 mL), filtered and rinsed with methyl tert-butyl ether (2 x 6 mL). The combined filtrates were concentrated under reduced pressure to give compound 25, which was used in the coupling reaction without further purification.

[0092]

[0093] Compounds 26-36 were synthesized using the same method.

[0094] Example 7

[0095] To a 10 mL reaction tube, nucleoside 37 (260 mg, 1 mmol) and 5.0 mL of anhydrous THF were added, and the mixture was cooled to 0°C in an ice-water bath. Tert-butylmagnesium chloride Grignard reagent (3.0 mL of a 1 M THF solution, 3.0 mmol) was added dropwise, and the reaction mixture was stirred at 0°C for 30 min. Subsequently, a solution of phosphorus reagent 2 (725 mg, 1.6 mmol) in 5 mL of THF was added dropwise at 0°C. The resulting clear reaction solution was warmed and stirred for 1 day. Saturated NH4Cl (15 mL) was added, stirred for 5 minutes, and the mixture was diluted with ethyl acetate (200 mL). The organic phase was separated, and the aqueous layer was extracted with ethyl acetate (50 mL). The combined organic layers were washed with water (30 mL), saturated NaHCO3 (2 x 30 mL), brine (40 mL), and dried over Na2SO4. The solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (0-10% methanol in dichloromethane) to obtain a white solid product II-1-1 (291 mg, yield 55%). LCMS-ESI + (m / z):530.2(M+H) + .

[0096] Example 8

[0097]

[0098] To a 10 mL reaction tube, nucleoside 37 (260 mg, 1 mmol) and 5.0 mL of anhydrous THF were added, and the mixture was cooled to 0°C in an ice-water bath. Tert-butylmagnesium chloride Grignard reagent (3.0 mL of a 1 M THF solution, 3.0 mmol) was added dropwise, and the reaction mixture was stirred at 0°C for 30 min. Subsequently, a solution of phosphorus reagent 3 (725 mg, 1.6 mmol) in 5 mL of THF was added dropwise at 0°C. The resulting clear reaction solution was warmed and stirred for 1 day. Saturated NH4Cl (15 mL) was added, stirred for 5 minutes, and the mixture was diluted with ethyl acetate (200 mL). The organic phase was separated, and the aqueous layer was extracted with ethyl acetate (50 mL). The combined organic layers were washed with water (30 mL), saturated NaHCO3 (2 x 30 mL), brine (30 mL), and dried over Na2SO4. The solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (0-10% methanol in dichloromethane) to obtain a white solid product II-1-2 (302 mg, yield 57%). LCMS-ESI + (m / z):530.2(M+H) + .

[0099] Example 9

[0100]

[0101] To a solution of nucleoside 37 (260 mg, 1.0 mmol) and N-methylimidazole (0.64 ml, 8 mmol) in dichloromethane (10 mL) was added dropwise a solution of compound 25 (1.14 g, 2 mmol) in DCM (10 mL) at 0°C. The addition was complete over approximately 1 hour. After the addition was complete, the reaction mixture was allowed to warm to room temperature overnight and then concentrated under reduced pressure to yield a yellow oil. The oil was diluted with EtOAc (200 mL) and water (100 mL). The organic layer was separated, washed with 5% aqueous ammonium chloride solution (2 x 60 mL) and 5% brine solution (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0-10% methanol in dichloromethane) to yield white solid products III-1-1 (222 mg, 28%) and III-1-2 (158 mg, 20%). LCMS-ESI + (m / z):793.3(M+H) + .

[0102] The following compounds were synthesized by similar methods:

[0103]

[0104]

[0105] Example 10 Anti-HBV activity determination

[0106] HepG2.2.15 cells overexpressing the HBV viral attachment receptor sodium-taurocholate cotransporting polypeptide (NTCP) were grown to confluence in DMEM growth medium (without sodium pyruvate, Life Technologies, Rockville, MD) supplemented with 10% FBS (Thermo Scientific, Waltham, MD), 1% penicillin / streptomycin (Life Technologies, Rockville, MD), and 2 mM L-glutamine (Life Technologies, Rockville, MD) in T175 flasks. Cells were infected with HBV AD38 viral particles (Texcell, Frederick, USA) at 4000 genome equivalents per cell. After allowing viral infection to occur for 4 days, infected cells were harvested from the flasks by trypsinization, washed twice with OptiMEM (Life Technologies, Rockville, MD), and resuspended at a density of 0.25E6 cells / ml in DMEM containing 2% FBS and 1% DMSO. Infected cells were seeded at a density of 20,000 cells / well in 384-well collagen-coated plates (Greiner, Austria) containing serially diluted compounds of the disclosure or DMSO (0.5%) in a final volume of 80 μl. The assay plates were incubated for 5 days, and the antiviral activity of the test compounds was determined by detecting the presence of HBV DNA in the culture supernatants using the QuantiGene™ 2.0 Nucleic Acid Quantification Kit (Affymetrix, Santa Clara, CA).

[0107] The culture supernatant was harvested and treated with a lysis buffer containing proteinase K (Affymetrix, Santa Clara, CA). The supernatant was incubated with an HBV viral DNA-specific probe (Affymetrix, Santa Clara, CA) at 55°C for 30 minutes. 0.2 M NaOH was then added at room temperature for 30 minutes to denature the DNA, followed by the addition of a neutralization buffer (Affymetrix, Santa Clara, CA). The resulting lysed and neutralized supernatant was then added to a QuantiGene™ 2.0 384-well plate coated with capture oligonucleotides and incubated at 55°C overnight. The HBV-specific probe set consisted of a Capture Extender oligonucleotide (CE) and a blocking probe. After the overnight incubation, the wells were incubated sequentially with a preamplifier, amplifier, and labeled probe coupled to alkaline phosphatase for 1 hour, with washes between incubations. After the final wash step, alkaline phosphatase substrate (Luminol APS5) was added and the resulting luminescent signal was read in an EnVision Multilabel Plate Reader (PerkinElmer, Santa Clara, CA). EC50 values ​​were calculated from the fit of the dose-response curves to a four-parameter equation. All EC50 values ​​represent the geometric mean of a minimum of four determinations. EC50 values ​​for certain compounds of the present disclosure are shown in Table 1. 50 The values ​​are reported in the table below.

[0108] For the cytotoxicity test, take one bottle of well-grown HepG2.2.15 cells, digest them with trypsin, and prepare a single-cell suspension. Count the cells using a cell counter and adjust the cell density to 2 × 105 cells / mL using DMEM medium containing 10% FBS serum. The cells are then seeded into a 96-well culture plate (100 μL per well). The cells are placed in a CO2 incubator at 5% CO2 and 37°C until 80% contact inhibition is achieved. The supernatant is aspirated and culture medium containing different concentration gradients of the test drug is added. Simultaneously, culture medium containing the corresponding concentration gradient of the positive control drug, telbivudine, is added. A cell blank control well is also set up. Three replicate wells are set up for each concentration gradient and cell blank control. Place in a CO2 incubator and incubate at 37°C for 72 hours. Add 10 μL of MTT (0.5 / L) to each well and continue incubating for another 4 hours. Then carefully remove the supernatant and add 150 μL of DMSO to each well. Shake to dissolve the formazan particles. Finally, use a microplate reader to compare the colorimetry, set the blank well to zero, and measure the A value at 546 nm. Calculate the cell viability and the half-toxic concentration (CC) based on the absorbance value. 50 , the results are shown in the table.

[0109] Anti-HBV activity test results of compounds

[0110] Compound <![CDATA[EC 50 (μM)]]> <![CDATA[HepG2 CC 50 (μM)]]> Compound <![CDATA[EC 50 (μM)]]> <![CDATA[CC 50 (μM)]]> II-1-1 0.31 >100 III-10-1 0.36 >100 II-1-2 0.53 >100 III-10-2 0.47 >100 II-2-1 0.48 >100 III-11-1 0.66 >100 II-2-2 0.61 >100 III-11-2 0.71 >100 II-3-1 0.38 >100 III-12-1 0.41 >100 II-3-2 0.45 >100 III-12-2 0.51 >100 II-4-1 0.33 >100 IV-1-1 0.33 >100 II-4-2 0.56 >100 IV-1-2 0.49 >100 II-5-1 0.47 >100 IV-2-1 0.32 >100 II-5-2 0.53 >100 IV-2-2 0.56 >100 II-6-1 0.71 >100 IV-3-1 0.44 >100 II-6-2 1.05 >100 IV-3-2 0.63 >100 III-1-1 0.29 >100 IV-4-1 0.41 >100 III-1-2 0.51 >100 IV-4-2 0.55 >100 III-2-1 0.35 >100 IV-5-1 0.44 >100 III-2-2 0.55 >100 IV-5-2 0.51 >100 III-3-1 0.66 >100 IV-6-1 0.38 >100 III-3-2 0.73 >100 IV-6-2 0.49 >100 III-4-1 0.34 >100 IV-7-1 0.46 89.3 III-4-2 0.45 >100 IV-7-2 0.73 92.5 III-5-1 0.41 >100 IV-8-1 0.51 >100 III-5-2 0.52 >100 IV-8-2 0.58 >100 III-6-1 0.44 >100 IV-9-1 0.35 >100 III-6-2 0.58 >100 IV-9-2 0.54 >100 III-7-1 0.83 85.6 IV-10-1 0.41 >100 III-7-2 0.69 76.8 IV-10-2 0.52 >100 III-8-1 0.40 >100 IV-11-1 0.79 87.5 III-8-2 0.52 >100 IV-11-2 1.33 >100 III-9-1 0.39 >100 IV-12-1 0.68 92.2 III-9-2 0.56 >100 IV-12-2 0.93 81.6 Clevudine It should be noted that in the above translation, there may be a small error in the original text where "III-5-2" is likely a typo and should be "III-5-2". The translation is adjusted accordingly. 0.30 >100

[0111] Example 11

[0112] In vivo PK test data of compounds

[0113] Hepatitis B virus (HBV) polymerase is responsible for HBV viral replication, and the substrates used for replication are nucleoside triphosphates. The compounds described in the present invention are phosphoramidate nucleotide analog prodrugs, a novel liver-targeting molecule designed to deliver the 5'-monophosphate of clavudine, which is then metabolized within hepatocytes to generate the active nucleoside triphosphate analog. Unlike other nucleotides, the active 5'-triphosphate of clavudine is a non-competitive, non-chain-terminating inhibitor of the active site polymerase of hepatitis B virus (HBV), thereby inhibiting polymerase activity and HBV viral replication. As a prodrug, the ability to generate the nucleoside triphosphate analog in vivo is directly correlated with its inhibitory activity against HBV polymerase. Therefore, the inventors conducted in vivo PK studies to test the ability of the compounds of the present invention to generate the active nucleoside triphosphate analog. The in vivo PK study model was an in vivo rat liver PK analysis.

[0114] Pharmacokinetic studies of the Example compounds' metabolism in rat liver to produce the active ingredient, clavudine triphosphate, were conducted in rats. Administration was by single oral gavage at a dose of 10 mg / 5 ml / kg. The Example compounds were formulated with 30% PEG-200 and 0.5% sodium carboxymethylcellulose. Liver samples were collected 1, 2, 4, 6, 12, and 24 hours after administration. For sampling, rats were sacrificed with CO2, and the livers were flushed with ice-cold saline via the hepatic portal vein. The livers were then cut into approximately 0.2-gram pieces, snap-frozen in liquid nitrogen, and stored at -80°C. Liver samples were analyzed for the active species, clavudine 5'-triphosphate, by LC-MS / MS.

[0115]

[0116] As can be seen from the test results of the diastereomers prepared in the examples, the concentration of clavudine triphosphate generated by the D-amino acid phosphate compound in the example compounds in hepatocytes is much higher than that of the L-amino acid phosphate isomer (ATI-2173), and also far exceeds the concentration of clavudine in the liver at the same dose, demonstrating extremely high clinical application value.

[0117] The above experimental results show that the compounds of the present invention have the ability to effectively inhibit HBV virus. Compared with the positive control compounds clavulanate or ATI-2173, they show significant superiority in both in vitro and in vivo activity evaluations. The compounds of the present invention do not show cytotoxicity within the tested concentration range and have good safety, and have excellent prospects for the treatment of HBV infection.

[0118] Although the present invention has been described in detail above, it will be understood by those skilled in the art that various modifications and variations of the present invention without departing from the spirit and scope of the present invention are within the scope of protection of the present invention. The scope of the present invention is not limited to the detailed description above, but should be attributed to the claims.

Claims

1. A nucleoside phosphoramidate compound having the following structure, and a salt thereof: 。 2. A pharmaceutical composition comprising an effective amount of the nucleoside phosphoramidate compound according to claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, excipient, or a combination thereof.

3. Use of the nucleoside phosphoramidate compound according to claim 1, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating diseases caused by HBV virus infection.

4. Use of the nucleoside phosphoramidate compound according to claim 1, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating diseases caused by HBV virus infection, characterized in that: The drug includes one or more agents selected from the following: HBV polymerase inhibitors, immunomodulators, interferons, pegylated interferons, viral fusion / entry inhibitors, viral maturation inhibitors, capsid assembly regulators, reverse transcriptase inhibitors, cyclophilin / TNF inhibitors, FXR agonists, TLR agonists, siRNA or ASO cccDNA inhibitors, gene silencing agents, HBx inhibitors, sAg secretion inhibitors and HBV vaccines, or a pharmaceutically acceptable salt of any one of the foregoing substances.

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