WO2018082503A1

WO2018082503A1 - Heterocyclic compound and preparation method and application thereof

Info

Publication number: WO2018082503A1
Application number: PCT/CN2017/108016
Authority: WO
Inventors: 蔡家强; 宋帅; 邓汉文; 曾宏; 宋宏梅; 唐祖建; 刘瑶; 田强; 黄海涛; 王利春; 王晶翼
Original assignee: 四川科伦博泰生物医药股份有限公司
Priority date: 2016-11-02
Filing date: 2017-10-27
Publication date: 2018-05-11
Also published as: CN108602842A; CN108602842B

Abstract

The present invention relates to a heterocyclic compound having anti-viral activity, a drug composition comprising same, a preparation method thereof, and an application thereof for the prevention or treatment of viral diseases including but not limited to viral hepatitis A, viral hepatitis B, viral hepatitis C, influenza, herpes, and acquired immunodeficiency syndrome (AIDS).

Description

Heterocyclic compound, preparation method and use thereof

Field of invention

The present invention relates to a heterocyclic compound having antiviral activity, a pharmaceutical composition comprising the same, a process for the preparation thereof, and its prevention or treatment including but not limited to hepatitis A virus, hepatitis B virus, hepatitis C virus, Use in influenza, herpes, and viral diseases of acquired immunodeficiency syndrome (AIDS).

Background of the invention

A virus consists of a nucleic acid molecule (DNA or RNA) or a protein (such as a prion). The virus can cause a variety of infectious diseases. Common diseases caused by viruses include, but are not limited to, viral hepatitis A, hepatitis B, hepatitis C, influenza, herpes and acquired immunodeficiency syndrome. (AIDS).

Currently used antiviral drugs play a role by inhibiting virus attachment, uncoating, viral gene duplication, maturation or release, or by affecting the host's immune system, including reverse transcriptase inhibitors and capsid protein assembly inhibitors. Wait.

Hepatitis B virus (HBV) is a common hepadnavirus-like viral pathogen. Such viruses can cause diseases such as acute hepatitis, chronic hepatitis, liver fibrosis, cirrhosis and liver cancer.

Hepatitis B can be treated with the following nucleoside analogues:

Among them, Tenofovir is a nucleoside analog reverse transcriptase inhibitor, which is effective against human immunodeficiency virus (HIV) and hepatitis B virus (HBV) infection (Antivir Ther, 2004, 9, 57- 65; J Viral Hepat, 2000, 7, 161-165). Its principle of action is to inhibit reverse transcriptase activity by competing with the natural substrate deoxyadenosine-5'-triphosphate to integrate DNA into HBV and then terminate DNA strand synthesis. But tenofovir is hardly absorbed by the gastrointestinal tract. The esterified prodrug tenofovir disoproxil fumarate (TDF) and the amidated prodrug tenofovir alafenamide fumarate (TAF) are well-soluble (Nucleosides Nucleotides Nucleic Acids. 2001, 20, 1085-1090; Antimicrob Agents Chemother. 2005, 49, 1898-1906; Antimicrob Agents Chemother. 2015, 59, 3563-3569; Journal of Hepatology. 2015, 62, 533-540).

In addition, hepatitis B can also be treated with non-nucleoside analogs. Deres et al. found that heteroaryldihydropyrimidines (Bay 41-4109) can block HBV virus replication by inhibiting viral capsid protein assembly (Science, 2003, 299, 893-896). Its specific mechanism of action is that dihydropyrimidines induce misassembly of core proteins, resulting in unstable capsid proteins that accelerate the degradation of core proteins (Biochem. Pharmacol., 2003, 66, 2273-2279). The heteroaryl dihydropyrimidine compound HAP1 (Proc. Natl. Acad. Sci., 2005, 102, 8138-8143) discovered by Zlotnick et al. and the heteroaryl dihydropyrimidine compound reported by Guangdong Dongyang Pharmaceutical Co., Ltd. (GLS4) (Antimicrob. Agents Chemother., 2013, 57, 5344-5354; WO2015078391, US2016206616) also has anti-HBV activity.

Although both of the above compounds exhibit antiviral activity to varying degrees, their activity has not been satisfactory. Therefore, the search for more effective drugs with less toxic side effects has urgent needs and important significance.

Summary of invention

The present invention provides an antiviral heterocyclic compound which exerts an antiviral action by inhibiting the assembly of reverse transcriptase and/or capsid protein. In particular, preferred compounds of the invention can simultaneously exhibit inhibition of reverse transcriptase and capsid protein assembly, thereby achieving superior antiviral effects at lower doses. In addition, the compounds of the invention also have better physicochemical properties (e.g., solubility, physical and/or chemical stability), improved pharmacokinetic properties (e.g., improved bioavailability, suitable half-life, and duration of action). , improved safety (lower toxicity and / or fewer side effects), less susceptible to drug resistance and other superior properties.

One aspect of the invention provides a compound or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph thereof Or a solvate, metabolite or prodrug, wherein the compound has the structure of formula (I):

among them:

B is selected from:

A is selected from C _1-6 alkylene, C _2-6 alkenylene, C _2-6 alkynylene, optionally substituted by one or more R ^b , said alkylene, alkenylene or sub An alkynyl group is optionally interrupted by one or more -O-, -NR- or -S-;

Or A is selected from the following groups:

among them

Represents a single bond or a double bond, and is connected to B at the 1 position and to the phosphorus atom (P) at the 2 position;

X, Y and Z are each independently selected from CH ₂ , O, S and NR at each occurrence;

R ^a and R ^b are each independently selected from the group consisting of halogen, -OH, -CN, -NO ₂ , -N(R) ₂ , -N ₃ , C _1-6 alkyl and C _3-6 ring at each occurrence. alkyl;

R ^{2 is} selected from the group consisting of H, C _1-12 alkyl, C _3-6 cycloalkyl, C _6-14 aryl, 5-14 membered heteroaryl, C _6-20 aralkyl, -C _1-6 Alkyl-COOH, -C _1-6 alkylene-C(=O)OC _1-6 alkyl, -C _1-6 alkylene-OC(=O)-C _1-6 alkyl, -C _1-6 alkylene-OC(=O)OC _1-6 alkyl and -((CH ₂ ) _i O) _m -(CH ₂ ) _q -OC _1-6 alkyl, each of the above groups optionally One or more selected from the group consisting of halogen, -OH, -CN, -NO ₂ , -N(R) ₂ , C _1-6 alkyl, halogenated C _1-6 alkyl, C _1-6 alkylthio Substituted with a substituent of a C _3-6 cycloalkyl group;

i and q are each independently 1, 2, 3, 4, 5 or 6 at each occurrence;

m is any integer selected from 0-50, preferably 0-20, particularly preferably 0-6;

R ^{1 is} selected from:

Representing a 3 to 14 membered nitrogen heterocycle, optionally additionally containing 1, 2 or 3 ring members independently selected from N, O, C=O, S, S=O and S(=O) ₂ ;

Ar ¹ and Ar ² are each independently selected from a C _6-14 aryl group and a 5-14 membered heteroaryl group, which are optionally selected from one or more selected from the group consisting of halogen, -OH, -CN, -NO ₂ , -N Substituent substitution of (R) ₂ , C _1-6 alkyl, halo C _1-6 alkyl, C _1-6 alkylthio and C _3-6 cycloalkyl;

L is absent or selected from -O-, -S-, and -NR-;

R ³ and R ⁴ are each independently selected from the group consisting of H, C _1-4 alkyl and C _3-6 cycloalkyl;

R ⁵ is attached to the remainder of the molecule with a single or double bond at each occurrence;

R ⁵ and R ⁶ are each independently selected from the group consisting of halogen, -OH, -COOH, -CN, -NO ₂ , -N(R) ₂ , C _1-6 alkyl, halogenated C _1-6 Alkyl, -WC _1-6 alkyl, -C _1-6 alkylene-WR, -WC _1-6 alkylene-W'-R, -WC _2-6 alkenyl, -C _2-6 Alkenyl-WR, -WC _2-6 alkenylene-W'-R and C _3-6 cycloalkyl, wherein the alkylene and alkenylene are optionally further separated by one or more W;

W and W' are each independently selected from the group consisting of O, C(=O), C(=O)O, NR, S, S=O, and S(=O) ₂ ;

R is each independently selected from the group consisting of H, C _1-6 alkyl and C _3-6 cycloalkyl;

n each independently is 0, 1, 2, 3, 4 or 5, with the proviso that n is not greater than the number of positions on the corresponding group that can be substituted;

When n is greater than 1, each R ^a may be the same or different, and each R ^b may be the same or different, and each R ⁵ may be the same or different, and each R ⁶ may be the same or different.

Another aspect of the invention provides a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound of the invention or a pharmaceutically acceptable compound thereof Accepted salts, esters, stereoisomers, tautomers, polymorphs, solvates, metabolites or prodrugs and one or more pharmaceutically acceptable carriers.

Another aspect of the invention provides a method of preparing a pharmaceutical composition comprising administering a compound of the invention, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph thereof, The solvate, metabolite or prodrug is combined with one or more pharmaceutically acceptable carriers.

Another aspect of the invention provides a compound of the invention, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, or the invention Use of a pharmaceutical composition for the preparation of a medicament for the prevention or treatment of a viral disease.

Another aspect of the invention provides a compound of the invention, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, or the invention A pharmaceutical composition for preventing or treating a viral disease.

Another aspect of the invention provides a method of preventing or treating a viral disease, the method comprising administering to an individual in need thereof an effective amount of a compound of the invention, or a pharmaceutically acceptable salt, ester, stereoisomer thereof, Tautomers, polymorphs, solvates, metabolites or prodrugs or pharmaceutical compositions of the invention.

The above viral diseases include, but are not limited to, viral hepatitis A, hepatitis B virus, hepatitis C virus, influenza, herpes, and acquired immunodeficiency syndrome (AIDS).

Another aspect of the invention provides a method of preparing a compound of the invention, the method comprising the steps of:

or

among them:

R ¹² , R ¹³ and R ¹⁴ are each independently selected from the group consisting of F, Cl, Br, I, -NHR, hydroxy, triflate, p-toluenesulfonate and borate;

PG is selected from the group consisting of tert-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, allyloxycarbonyl, trimethylsilyloxycarbonyl, methoxycarbonyl, ethoxycarbonyl, p-toluenesulfonyl, o-nitrobenzenesulfonate Acyl, p-nitrophenylsulfonyl, formyl, acetyl, trifluoroacetyl, propionyl, pivaloyl, phenyl, benzoyl, trityl, benzyl, 2,4-dimethoxy Benzyl and p-methoxybenzyl;

The remaining groups are as defined above;

The first step is carried out in the presence of an organic base or an inorganic base and/or a condensation reagent (particularly preferably DCC, DIC, EDC, BOP, PyAOP or PyBOP) in an aprotic solvent or without solvent;

The second step is carried out under conditions suitable for removal of the PG group;

The third step is carried out in an aprotic solvent in the presence of an organic or inorganic base;

The fourth step is carried out in an aprotic solvent, preferably in the presence of an organic base or an inorganic base and/or a condensation reagent, particularly preferably DCC, DIC, EDC, BOP, PyAOP or PyBOP.

among them:

R ^{12 is} selected from the group consisting of F, Cl, Br, I, -NHR, hydroxy, triflate, p-toluenesulfonate, and borate;

LG is a leaving group, which is preferably a pentafluorophenyl group and a p-nitrophenyl group;

The remaining groups are as defined above;

The first step is carried out in an aprotic solvent in the presence of an organic or inorganic base;

The second step is carried out in an aprotic solvent in the presence of an organic or inorganic base;

The third step is carried out in an aprotic solvent in the presence of an organic or inorganic base.

definition

Unless otherwise defined below, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. References to techniques used herein are intended to refer to techniques that are generally understood in the art, including those that are obvious to those skilled in the art. While the following terms are believed to be well understood by those skilled in the art, the following definitions are set forth to better explain the invention.

The terms "comprising," "comprising," "having," "containing," Or method steps.

The term "alkylene" as used herein denotes a saturated divalent hydrocarbon group, preferably a saturated divalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms, such as methylene, ethylene, Propylene or butylene.

The term "alkenylene" as used herein denotes a divalent hydrocarbon radical containing one or more double bonds, preferably having 2, 3, 4, 5 or 6 carbon atoms, such as ethenylene, propenylene or Allylene. When the compounds of the invention contain alkenylene groups, the compounds may exist in pure E (entgegen) form, pure Z (zusammen) form, or any mixture thereof.

The term "alkynylene" as used herein denotes a divalent hydrocarbon radical containing one or more triple bonds, preferably having 2, 3, 4, 5 or 6 carbon atoms, for example ethynylene or propynylene. .

As used herein, the term "alkyl" is defined as a straight or branched saturated aliphatic hydrocarbon group. In some embodiments, an alkyl group has from 1 to 12 carbon atoms, such as from 1 to 6 carbon atoms. For example, as used herein, the term " _C1-6 alkyl" refers to a straight or branched saturated aliphatic hydrocarbon group having from 1 to 6 carbon atoms (eg, methyl, ethyl, n-propyl, isopropyl) , n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl or n-hexyl), which are optionally substituted by one or more (such as 1 to 3) suitable substituents such as halogen (this This group is referred to as "haloalkyl" (for example, CF ₃ , C ₂ F ₅ , CHF ₂ , CH ₂ F, CH ₂ CF ₃ , CH ₂ Cl or -CH ₂ CH ₂ CF _{3 ,} etc.). The term "C _1-4 alkyl" refers to a straight or branched saturated aliphatic hydrocarbon group having 1 to 4 carbon atoms (ie, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl) , sec-butyl or tert-butyl).

As used herein, the term "cycloalkyl" refers to a saturated or unsaturated, non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (eg, a monocyclic ring such as cyclopropyl, cyclobutyl, cyclopentyl, Cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, or bicyclic, including spiro, fused or bridged systems (such as bicyclo [1.1.1] pentyl, bicyclo [2.2.1] heptyl, bicyclo [ 3.2.1] Octyl or bicyclo [5.2.0] anthracenyl, decalinyl, etc.)), which is optionally substituted by one or more (such as 1 to 3) suitable substituents. The cycloalkyl group has 3 to 15 carbon atoms. For example, the term "C _3-6 cycloalkyl" refers to a saturated or unsaturated, non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring having from 3 to 6 ring-forming carbon atoms (eg, cyclopropyl, cyclobutyl) A group, a cyclopentyl group or a cyclohexyl group, which is optionally substituted by one or more (such as 1 to 3) suitable substituents, such as a methyl-substituted cyclopropyl group.

As used herein, the term "aryl" refers to an all-carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated pi-electron system. For example, as used herein, the term " _C6-14 aryl" means an aromatic group containing from 6 to 14 carbon atoms, such as phenyl or naphthyl. The aryl group is optionally substituted by one or more (such as 1 to 3) suitable substituents (e.g., halogen, -OH, -CN, -NO ₂ , C _1-6 alkyl, etc.).

The term "aralkyl" as used herein denotes an alkyl group substituted with an aryl group, wherein the aryl group and the alkyl group are as defined herein. Typically, the aryl group can have from 6 to 14 carbon atoms and the alkyl group can have from 1 to 6 carbon atoms. Exemplary aralkyl groups include, but are not limited to, benzyl, phenylethyl, phenylpropyl, phenylbutyl.

The term "heteroaryl" as used herein refers to a monocyclic, bicyclic or tricyclic aromatic ring system having 5, 6, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 1 or 2 or 3 or 4 or 5 or 6 or 9 or 10 carbon atoms, and which contain at least one hetero atom which may be the same or different (the hetero atom is, for example, oxygen, nitrogen or sulfur), and additionally In one case, it may be benzo-fused. In particular, the heteroaryl is selected from the group consisting of thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thia A oxazolyl group or the like, and a benzo derivative thereof; or a pyridyl group, a pyridazinyl group, a pyrimidinyl group, a pyrazinyl group, a triazinyl group or the like, and a benzo derivative thereof.

The term "halo" or "halogen" group, as used herein, is defined to include F, Cl, Br or I.

As used herein, the term "alkylthio" refers to an alkyl group, as defined above, appended to the parent molecular moiety through a sulfur atom. Representative examples of _C1-6 alkylthio include, but are not limited to, methylthio, ethylthio, tert-butylthio, and hexylthio.

As used herein, the term "nitrogen heterocycle" refers to a saturated or unsaturated monocyclic or bicyclic group having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 in the ring. , 12 or 13 carbon atoms and at least one nitrogen atom, which may also optionally comprise one or more (eg one, two, three or four) selected from the group consisting of N, O, C=O, S, S a ring member of =O and S(=O) ₂ ; the nitrogen heterocycle is attached to the remainder of the molecule through a nitrogen atom. In particular, the 3 to 14 membered nitrogen heterocycle is a cyclic group having 3 to 14 carbon atoms and a hetero atom (at least one of which is a nitrogen atom) in the ring, including but not limited to aziridine, nitrogen Heterocyclic butane, pyrrolyl, pyrrolidinyl, pyrrolinyl, pyrrolidinyl, imidazolyl, imidazolidinyl, imidazolinyl, pyrazolyl, pyrazolinyl, piperidinyl, morpholinyl, sulfur? A phenyl group, a piperazinyl group, a fluorenyl group, a porphyrin group or the like.

The term "substituted" means that one or more (eg, one, two, three or four) hydrogens on the designated atom are replaced by the selection of the indicated group, provided that the specified atom is not present at present. The normal valence in the case and the substitution form a stable compound. Combinations of substituents and/or variables are permissible only if such combinations form stable compounds.

If a substituent is described as "optionally substituted," the substituent may be unsubstituted or (2) substituted. If the carbon of the substituent is described as being optionally substituted by one or more of the list of substituents, then one or more hydrogens on the carbon (to the extent of any hydrogen present) may be independently and/or together independently The optional substituents selected are substituted. If the nitrogen of the substituent is described as being optionally substituted by one or more of the list of substituents, then one or more hydrogens on the nitrogen (to the extent of any hydrogen present) may each be independently selected. Substitute substitution.

If a substituent is described as being "independently selected from" a group, each substituent is selected independently of the other. Thus, each substituent may be the same or different from another (other) substituent.

As used herein, the term "one or more" means 1 or more than 1, such as 2, 3, 4, 5 or 10 under reasonable conditions.

Unless otherwise indicated, a point of attachment of a substituent, as used herein, may come from any suitable position of the substituent.

When a bond of a substituent is shown to pass through a bond connecting two atoms in the ring, such a substituent may be bonded to any of the ring-forming atoms in the substitutable ring.

The invention also includes all pharmaceutically acceptable isotopically-labeled compounds which are identical to the compounds of the invention, except that one or more atoms are of the same atomic number but the atomic mass or mass number differs from the atomic mass prevailing in nature. Or atomic substitution of mass. Examples of isotopes suitable for inclusion in the compounds of the invention include, but are not limited to, isotopes of hydrogen (e.g., ² H, ³ H); isotopes of carbon (e.g., ¹¹ C, ¹³ C, and ¹⁴ C); isotopes of chlorine (e.g. ³⁶ Cl); isotope of fluorine (eg ¹⁸ F); isotopes of iodine (eg ¹²³ I and ¹²⁵ I); isotopes of nitrogen (eg ¹³ N and ¹⁵ N); isotopes of oxygen (eg ¹⁵ O, ¹⁷ O and ¹⁸ O) ); an isotope of phosphorus (eg, ³² P); and an isotope of sulfur (eg, ³⁵ S). Certain isotopically-labeled compounds of the invention (e.g., those incorporating radioisotopes) are useful in drug and/or substrate tissue distribution studies (e.g., assays). The radioisotope ruthenium (i.e., ³ H) and carbon-14 (i.e., ¹⁴ C) are particularly useful for this purpose because of their ease of incorporation and ease of detection. Substitution with positron emitting isotopes (eg, ¹¹ C, ¹⁸ F, ¹⁵ O, and ¹³ N) can be used to examine substrate receptor occupancy in positron emission tomography (PET) studies. Isotopically labeled compounds of the invention can be prepared by replacing the previously employed non-labeled reagents with suitable isotopically labeled reagents by methods analogous to those described in the accompanying routes and/or examples and preparations. The pharmaceutically acceptable solvates of the present invention include those in which the crystallization solvent can be substituted with an isotope, for example, D ₂ O, acetone-d ₆ or DMSO-d ₆ .

The term "stereoisomer" denotes an isomer formed by at least one asymmetric center. In a compound having one or more (eg, one, two, three or four) asymmetric centers, it can produce a racemic mixture, a single enantiomer, a mixture of diastereomers, and Diastereomers. Specific individual molecules can also exist as geometric isomers (cis/trans). Similarly, the compounds of the invention may exist as mixtures (often referred to as tautomers) of two or more different forms in a rapidly balanced structure. Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers Wait. For example, a dihydropyrimidinyl group can exist in equilibrium in the following tautomeric forms:

It is to be understood that the scope of the present application covers all such ratios in any ratio (eg, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99). %) isomer or a mixture thereof.

Solid lines can be used in this article

Solid wedge

Virtual wedge

The carbon-carbon bonds of the compounds of the invention are depicted. The use of solid lines to delineate linkages bonded to an asymmetric carbon atom is intended to include all possible stereoisomers at the carbon atom (eg, specific enantiomers, racemic mixtures, etc.). The use of a solid or virtual wedge to characterize a bond to an asymmetric carbon atom is intended to indicate the presence of the stereoisomers shown. When present in a racemic mixture, solid and virtual wedges are used to define relative stereochemistry rather than absolute stereochemistry. Unless otherwise indicated, the compounds of the invention are intended to be stereoisomers (including cis and trans isomers, optical isomers (eg, R and S enantiomers), diastereomers, Geometric isomers, rotamers, conformers, atropisomers, and mixtures thereof exist. The compounds of the invention may exhibit more than one type of isomerism and consist of a mixture thereof (e.g., a racemic mixture and a diastereomeric pair).

The invention encompasses all possible crystalline forms or polymorphs of the compounds of the invention, which may be a single polymorph or a mixture of more than one polymorph in any ratio.

It will also be understood that certain compounds of the invention may exist in free form for treatment or, where appropriate, in the form of their pharmaceutically acceptable derivatives. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, metabolites or prodrugs, which are capable of being administered directly to a patient in need thereof The compound of the invention or a metabolite or residue thereof is provided indirectly or indirectly. Thus, when reference is made herein to a "compound of the invention," it is also intended to encompass the various derivative forms described above for the compound.

The pharmaceutically acceptable salts of the compounds of the present invention include the acid addition salts and base addition salts thereof.

Suitable acid addition salts are formed from acids which form pharmaceutically acceptable salts. Examples include aspartate, bicarbonate/carbonate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hydrobromide/bromide, hydrogen Iodate/iodide, naphthylate, nicotinate, nitrate, orotate, palmitate and other similar salts.

Suitable base addition salts are formed from bases which form pharmaceutically acceptable salts. Examples include aluminum salts, arginine salts, choline salts, lysine salts, magnesium salts, meglumine salts, tromethamine salts, and other similar salts.

For a review of suitable salts, see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the invention are known to those skilled in the art.

As used herein, the term "ester" means an ester derived from a compound of the formulae herein, which includes a physiologically hydrolyzable ester (which can be hydrolyzed under physiological conditions to release the free acid or alcohol form of the invention). Compound). The compounds of the invention may also be esters per se.

The compound of the present invention may exist in the form of a solvate (preferably a hydrate) wherein the compound of the present invention contains a polar solvent as a structural element of the crystal lattice of the compound, particularly such as water, methanol or ethanol. The amount of polar solvent, particularly water, may be present in stoichiometric or non-stoichiometric ratios.

Also included within the scope of the invention are metabolites of the compounds of the invention, i.e., substances formed in vivo upon administration of a compound of the invention. Such products may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, delipidization, enzymatic hydrolysis, and the like of the administered compound. Accordingly, the invention includes metabolites of the compounds of the invention, including compounds prepared by contacting a compound of the invention with a mammal for a time sufficient to produce a metabolic product thereof.

The invention further includes within its scope prodrugs of the compounds of the invention which are certain derivatives of the compounds of the invention which may themselves have less or no pharmacological activity, when administered to or into the body It can be converted to a compound of the invention having the desired activity by, for example, hydrolytic cleavage. Typically such prodrugs will be functional group derivatives of the compounds which are readily converted in vivo to the desired therapeutically active compound. Additional information on the use of prodrugs can be found in "Pro-drugs as Novel Delivery" Systems, Vol. 14, ACS Symposium Series (T. Higuchi and V. Stella) and "Bioreversible Carriers in Drug Design," Pergamon Press, 1987 (EB Roche, ed., American Pharmaceutical Association). Prodrugs of the invention can be passed, for example, Substituting certain parts of the compounds of the invention as known to the "pro-moiety" (for example "Design of Prodrugs", H. Bundgaard (Elsevier, 1985)" by those skilled in the art Prepare with appropriate functional groups.

The invention also encompasses compounds of the invention containing a protecting group. In any process for preparing a compound of the invention, it may be necessary and/or desirable to protect a sensitive group or reactive group on any of the molecules of interest, thereby forming a chemically protected form of the compound of the invention. This can be achieved by conventional protecting groups such as those described in Protective Groups in Organic Chemistry, ed. JFW McOmie, Plenum Press, 1973; and TW Greene & P. GM Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991. Protecting groups, which are incorporated herein by reference. The protecting group can be removed at a suitable subsequent stage using methods known in the art.

The term "about" means within ±10% of the stated value, preferably within ±5%, more preferably within ±2%.

Detailed description of the invention

Compound and preparation method thereof

In one embodiment, the invention provides a compound, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein The compound has the structure of formula (I):

among them:

B is selected from:

Or A is selected from the following groups:

among them

i and q are each independently 1, 2, 3, 4, 5 or 6 at each occurrence;

R ^{1 is} selected from:

L is absent or selected from -O-, -S-, and -NR-;

n each independently is 0, 1, 2, 3, 4 or 5, with the proviso that n is not greater than the number of positions on the corresponding group that can be substituted, and

In a preferred embodiment, X, Y, and Z at each occurrence is independently selected from CH _2, O, and NH.

In a particularly preferred embodiment, X, Y and Z are each independently selected from O and NH at each occurrence.

In a preferred embodiment, R ^a and R ^b are each independently selected from the group consisting of halogen, -OH, -N(R) ₂ , -N ₃ and C _1-6 alkyl.

In a particularly preferred embodiment, R ^a and R ^b are each independently selected from the group consisting of F, -OH, amino, cyclopropylamino and methyl at each occurrence.

In a preferred embodiment, B is selected from the group consisting of:

In a particularly preferred embodiment, B is

In a preferred embodiment, A is selected from the group consisting of:

One of the positions is connected to B, and the second position is connected to the phosphorus atom (P).

In a more preferred embodiment, A is

In a particularly preferred embodiment, A is

In a preferred embodiment,

Represents a 3-, 4-, 5-, or 6-membered nitrogen heterocycle.

In a particularly preferred embodiment,

It is selected from the group consisting of azetidinyl, pyrrolidinyl, piperidinyl, morpholinyl and piperazinyl.

In a preferred embodiment, R ^{1 is} selected from the group consisting of

In a preferred embodiment, Ar ¹ and Ar ² are each independently selected from the group consisting of imidazolyl, oxazolyl, thiazolyl, pyridyl, pyrimidinyl, piperazinyl and phenyl, each of which is optionally one or more The same or different halogen, C _1-6 alkyl, halo C _1-6 alkyl and C _3-6 cycloalkyl are substituted.

In a more preferred embodiment, Ar ¹ and Ar ² are each independently selected from the group consisting of imidazolyl, oxazolyl, thiazolyl, pyridyl, pyrimidinyl, piperazinyl and phenyl, each of which is optionally one or more The same or different halogen, C _1-6 alkyl and C _3-6 cycloalkyl substituted.

In a preferred embodiment, Ar ^{1 is} selected from the group consisting of

Wherein R ^c is each independently selected from the group consisting of F, Cl, Br, I, C _1-6 alkyl, halo C _1-6 alkyl, and C _3-6 cycloalkyl;

Preferably, each occurrence of R ^c is independently selected from the group consisting of F, Cl, Br, I, C _1-6 alkyl and C _3-6 cycloalkyl;

Ar ¹ is preferably selected from:

Ar ^{1 is} more preferably from:

In a preferred embodiment, Ar ^{2 is} selected from the group consisting of

The above groups are optionally substituted by one or more groups selected from the group consisting of halogen, _C1-6 alkyl, halogenated _C1-6 alkyl and _C3-6 cycloalkyl.

In a more preferred embodiment, Ar ^{2 is} selected from the group consisting of

The above groups are optionally substituted by one or more groups selected from the group consisting of halogen, C _1-6 alkyl and C _3-6 cycloalkyl.

In a preferred embodiment, L is -O-.

In a preferred embodiment, R ³ and R ⁴ are each independently selected from the group consisting of methyl, ethyl, n-propyl and isopropyl.

In a particularly preferred embodiment, R ⁴ is H.

In a preferred embodiment, each occurrence of R ⁵ and R ⁶ is independently selected from the group consisting of F, Cl, Br, I, -OH, -COOH, methyl, ethyl, -CH ₂ OH, -OCH ₃ , -CH ₂ COOH, -CH ₂ CH ₂ COOH, -CH ₂ CH ₂ CH ₂ COOH, -CH=CHCOOH, -OCH ₂ COOH, -SCH ₂ COOH, -N(CH ₃ )CH ₂ COOH, -CH ₂ OCH ₂ COOH, -CH ₂ SCH ₂ COOH and -CH ₂ N(CH ₃ )CH ₂ COOH.

In a preferred embodiment, R ^{2 is} selected from the group consisting of H, C _1-12 alkyl, C _3-6 cycloalkyl, C _6-14 aryl, 5-14 membered heteroaryl, C _6-20 aralkyl , -C _1-6 alkylene-C(=O)OC _1-6 alkyl, -C _1-6 alkylene-OC(=O)-C _1-6 alkyl, -C _1-6 Alkyl-OC(=O)OC _1-6 alkyl and -((CH ₂ ) _i O) _m -(CH ₂ ) _q -OC _1-6 alkyl, each of which is optionally one or more One selected from the group consisting of halogen, -OH, -CN, -NO ₂ , -N(R) ₂ , C _1-6 alkyl, halogenated C _1-6 alkyl, C _1-6 alkylthio and C _3- a substituent of a ₆ cycloalkyl group;

In a more preferred embodiment, R ^{2 is} selected from the group consisting of: C _1-6 alkyl, -((CH ₂ ) _i O) _m -(CH ₂ ) _q -OC _1-6 alkyl,

among them:

i and q are each independently 1, 2, 3, 4, 5 or 6 at each occurrence;

R ⁷ , R ⁸ and R ⁹ are each independently selected from the group consisting of H, C _1-10 alkyl, C _3-6 cycloalkyl, C _6-20 aryl and C _7-20 aralkyl, said alkyl group, The cycloalkyl, aryl and aralkyl groups are each optionally substituted by one or more substituents selected from the group consisting of halogen, -OH, -CN and -NO ₂ ;

Or R ⁷ and R ⁸ together with the carbon atom to which they are attached form a C _3-6 cycloalkyl group;

R ¹⁰ and R ¹¹ are each independently selected from the group consisting of H, halogen, -OH, -CN, -NO ₂ , C _1-10 alkyl, and C _3-6 cycloalkyl.

In a more preferred embodiment, R ^{2 is} selected from the group consisting of: C _1-6 alkyl, -((CH ₂ ) _i O) _m -(CH ₂ ) _q -OC _1-4 alkyl,

among them:

i and q are each independently 1, 2, 3 or 4 at each occurrence;

m is 1, 2, 3, 4, 5 or 6;

R ⁷ and R ⁹ are each independently selected from H or C _1-4 alkyl, and the alkyl group is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -CN, and -NO ₂ ;

R ¹⁰ and R ¹¹ are each independently selected from H, halogen, -OH, -CN, -NO ₂ or C _1-4 alkyl.

In a particularly preferred embodiment, R ^{2 is} selected from the group consisting of H, methyl, ethyl, propyl, butyl, pentyl, -((CH ₂ ) ₂ O) ₆ -(CH ₂ ) ₂ -O-CH ₃ ,

In a particularly preferred embodiment, R ^{2 is} selected from the group consisting of methyl, ethyl, propyl, butyl, -((CH ₂ ) ₂ O) ₆ -(CH ₂ ) ₂ -O-CH ₃ ,

In a preferred embodiment, n is 0, 1, 2 or 3.

In a preferred embodiment, the invention provides a compound, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein The compound has the structure of formula (II) or formula (II)-1:

In a preferred embodiment, the invention provides a compound, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein The compound has any structure of the following formula:

The present invention encompasses compounds obtained by any combination of the various embodiments.

In a preferred embodiment, the invention provides a compound, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein Said compound is selected from:

One embodiment of the invention provides a method of preparing a compound of the invention, the method comprising the steps of:

or

among them:

The remaining groups are as defined above;

The first step is carried out in the presence of an organic base or an inorganic base and/or a condensation reagent in an aprotic solvent or without a solvent;

The fourth step is carried out in an aprotic solvent, preferably in the presence of an organic base or an inorganic base and/or a condensation reagent.

among them:

The remaining groups are as defined above;

In a preferred embodiment, R ¹² , R ¹³ and R ¹⁴ are each independently selected from the group consisting of F, Cl, Br, I, NH ₂ or a hydroxyl group.

In a preferred embodiment, PG may be a tert-butoxycarbonyl group, and the conditions suitable for the removal of the tert-butoxycarbonyl group may be, for example, catalyzed by an acid such as trifluoroacetic acid in a solvent such as dichloromethane at 0. °C to room temperature.

In a preferred embodiment, LG-OH can be pentafluorophenol, p-nitrophenol.

In a preferred embodiment, the aprotic solvent which can be used in the method of preparing the compound of the present invention includes, but is not limited to, tetrahydrofuran, dichloromethane, 1,2-dichloroethane, acetonitrile, N,N-dimethyl Formamide (DMF), N-methylpyrrolidone, 1,3-dimethyl-2-imidazolidinone (DMI), dimethyl sulfoxide (DMSO), and hexamethylphosphoric triamide (HMPA).

In a preferred embodiment, the organic bases which can be used in the preparation of the compounds of the invention include, but are not limited to, sodium t-butoxide, t-butylmagnesium chloride, triethylamine, N,N-diisopropylethylamine (DIPEA) , pyridine or 4-dimethylaminopyridine (DMAP); inorganic bases which can be used in the preparation of the compounds of the invention include, but are not limited to, NaH, NaOH, Na ₂ CO ₃ or K ₂ CO ₃ .

In a preferred embodiment, the condensation reagents which can be used in the method of preparing the compounds of the invention include, but are not limited to, dicyclohexylcarbodiimide (DCC), N,N-diisopropylcarbodiimide (DIC). , 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate ( BOP), (3H-1,2,3-triazolo[4,5-b]pyridin-3-yloxy)tri-1-pyrrolidinylphosphonium hexafluorophosphate (PyAOP) and 1H-benzo Triazol-1-yloxytripyrrolidinylphosphonium hexafluorophosphate (PyBOP).

Pharmaceutical compositions and methods of treatment

In another embodiment, the invention provides a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph thereof, or a pharmaceutically acceptable salt thereof A form, solvate, metabolite or prodrug and one or more pharmaceutically acceptable carriers. In another embodiment, the pharmaceutical composition may further comprise one or more additional therapeutic agents, such as other therapeutic agents for preventing or treating viral diseases.

In another embodiment, the invention provides a method of preparing a pharmaceutical composition comprising the compound of the invention, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph thereof, or a pharmaceutically acceptable salt thereof The form, solvate, metabolite or prodrug is combined with one or more pharmaceutically acceptable carriers.

In another embodiment, the invention provides a compound of the invention, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, Or the use of the pharmaceutical composition of the present invention in the preparation of a medicament for preventing or treating a viral disease.

In another embodiment, the invention provides a compound of the invention, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, Or a pharmaceutical composition of the invention for use in the prevention or treatment of a viral disease.

In another embodiment, the invention provides a method of preventing or treating a viral disease, the method comprising administering to an individual in need thereof an effective amount of a compound of the invention, or a pharmaceutically acceptable salt, ester, or stereoisod thereof A construct, tautomer, polymorph, solvate, metabolite or prodrug or a pharmaceutical composition of the invention.

Preferred compounds of the invention exert an antiviral effect by inhibiting the assembly of reverse transcriptase and/or capsid protein. Thus, the compounds of the invention are useful in the treatment of any virus involved in the assembly of reverse transcriptase and/or capsid proteins in the process of affecting a host, including but not limited to hepatitis A virus (HAV), hepatitis B virus (HBV) ), hepatitis C virus (HCV), influenza virus, herpes virus (HSV) and human immunodeficiency virus (HIV).

Thus, viral diseases which can be prevented and treated using the compounds of the present invention include, but are not limited to, viral hepatitis A, hepatitis B virus, hepatitis C virus, influenza, herpes, and acquired immunodeficiency syndrome. (AIDS), and related symptoms or diseases caused by the above diseases (for example, inflammation, liver fibrosis, cirrhosis, liver cancer, etc.).

"Pharmaceutically acceptable carrier" in the context of the present invention means a diluent, adjuvant, excipient or vehicle with which the therapeutic agent is administered, and which is suitable for contacting humans and/or within the scope of sound medical judgment. Tissues of other animals without excessive toxicity, irritation, allergic reactions, or other problems or complications corresponding to a reasonable benefit/risk ratio.

Pharmaceutically acceptable carriers that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, sterile liquids such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, minerals. Oil, sesame oil, etc. Water is an exemplary carrier when the pharmaceutical composition is administered intravenously. It is also possible to use physiological saline and an aqueous solution of glucose and glycerin as a liquid carrier, particularly for injection. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, maltose, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skimmed milk powder, glycerin, propylene glycol, water, Ethanol and the like. The composition may also contain minor amounts of wetting agents, emulsifying agents or pH buffering agents as needed. Oral formulations may contain standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like. Examples of suitable pharmaceutically acceptable carriers are as described in Remington's Pharmaceutical Sciences (1990).

The pharmaceutical compositions of the invention may act systemically and/or locally. For this purpose, they may be administered in a suitable route, for example by injection (for example intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular, including instillation) or transdermal administration; or by oral, buccal, or oral administration. Nasal, transmucosal, topical, in the form of ophthalmic preparations or by inhalation.

For these routes of administration, the pharmaceutical compositions of the invention may be administered in a suitable dosage form.

The dosage forms include, but are not limited to, tablets, capsules, troches, hard candy, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions. Injectable solutions, elixirs, syrups.

The term "effective amount" as used herein refers to an amount of a compound that, to a certain extent, relieves one or more symptoms of the condition being treated after administration.

The dosing regimen can be adjusted to provide the optimal desired response. For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the urgent need for treatment. It is noted that the dose value can vary with the type and severity of the condition to be alleviated and can include single or multiple doses. It is to be further understood that for any particular individual, the particular dosage regimen will be adjusted over time according to the individual needs and the professional judgment of the person administering the composition or the composition of the supervised composition.

The amount of the compound of the invention administered will depend on the severity of the individual, condition or condition being treated, the rate of administration, the handling of the compound, and the judgment of the prescribing physician. Generally, an effective dose will be from about 0.0001 to about 50 mg per kg body weight per day, for example from about 0.01 to about 10 mg/kg/day (single or divided doses). For a 70 kg person, this would add up to about 0.007 mg/day to about 3500 mg/day, such as from about 0.7 mg/day to about 700 mg/day. In some cases, a dose level that is not higher than the lower limit of the aforementioned range may be sufficient, while in other cases, a larger dose may still be employed without causing any harmful side effects, provided that the larger The dose is divided into several smaller doses to be administered throughout the day.

The amount or amount of the compound of the present invention in the pharmaceutical composition may be from about 0.01 mg to about 1000 mg, suitably from 0.1 to 500 mg, preferably from 0.5 to 300 mg, more preferably from 1 to 150 mg, particularly preferably from 1 to 50 mg, for example, 1.5 mg, 2 mg, 4 mg, 10 mg, 25 mg, and the like.

The term "treating" as used herein, unless otherwise indicated, means reversing, alleviating, inhibiting the progression of a condition or condition to which such a term applies or one or more symptoms of such a condition or condition, or Prevention of such a condition or condition or one or more symptoms of such condition or condition.

"Individual" as used herein includes human or non-human animals. Exemplary human individuals include a human individual (referred to as a patient) or a normal individual having a disease, such as the disease described herein. "Non-human animals" in the present invention include all vertebrates, such as non-mammals (eg, birds, amphibians, reptiles) and mammals, such as non-human primates, domestic animals, and/or domesticated animals (eg, sheep, dogs). , cats, cows, pigs, etc.).

In another embodiment, the pharmaceutical compositions of the present invention may further comprise one or more additional therapeutic or prophylactic agents including, but not limited to, lamivudine, telbivudine, entecavir, adefovir Ester, tenofovir, tenofovir disoproxil fumarate, and tenofovir alafenamide fumarate.

Example

The invention is further described in the following examples, but these examples are not intended to limit the scope of the invention.

Commercially available anhydrous solvents and HPLC grade solvents were used without further purification unless otherwise stated.

¹ H NMR spectra were recorded at ambient temperature using a Bruker instrument (400 MHz) using TMS as an internal standard. The chemical shift (δ) is given in ppm and the coupling constant (J) is given in hertz (Hz). ^The fractional weights of the ¹ H NMR spectrum peaks are abbreviated as follows: s (single peak), d (doublet), t (triplet), q (quadruple), m (multiplet), br (broad).

LC-MS was detected on an Aglient 1200 liquid chromatograph using an Aglient 6120 Quadrupole mass spectrometer at 214 nm and 254 nm. Prepare liquid chromatography using SHIMADZU CBM-20A and Aglient 1260 to prepare liquid phase meter, C18OBD 19×150mm 5μM preparative column, detection wavelength 214nm, mobile phase A is water, mobile phase B is acetonitrile (add 0.5 ‰ formic acid), press The following table performs a linear gradient elution:

时间(min)Time (min)	A％A%	B％B%
00	9090	1010
1515	4040	6060
3030	1010	9090

Example 1 (4R)-6-((4-(4-(((((()))))))))) ((2-methylbenzyl)oxy)phosphonyloxy)phenyl)piperidin-1-yl)methyl)-4-(2-chloro-4-fluorophenyl)-2- Ethyl (thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate (Compound 1)

Step 1: Synthesis of 4-(4-hydroxyphenyl)piperidine-1-carboxylic acid tert-butyl ester (Compound 1-2)

4-(4-(Benzyloxy)phenyl)-3,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (Compound 1-1) (3.0 g, 8.2 mmol) was added at room temperature After the compound was completely dissolved in tetrahydrofuran (10 mL) and methanol (20 mL), 10% Pd/C (300 mg) was added under nitrogen atmosphere, and the mixture was replaced with hydrogen. The mixture was reacted at room temperature for 18 hours, filtered, and the filtrate was post-treated. The title compound (2.0 g). ESI-MS (m/z): 2221. [M-55+H] ⁺ .

Step 2: 4-(4-((((()))))))))))))))))) Synthesis of tert-butyl ester of oxy)phenyl)piperidine-1-carboxylate (Compound 1-3)

(R)-9-(2-methoxymethoxypropyl)-adenine (3.1 g, 10.8 mmol) and compound 1-2 (2.0 g, 7.2 mmol) were dissolved in N-methylpyrrolidone at room temperature ( In 20 mL), the temperature was raised to 85 ° C, and triethylamine (879 mg, 8.9 mmol) was added dropwise. After the addition, the temperature was further increased to 100 ° C, and dicyclohexylcarbodiimide (4.8 g, 23.1 mmol) of N-A was added dropwise. A solution of the pyrrolidone (10 mL) was added dropwise, and the reaction was carried out at 100 ° C overnight to give the title compound (1.0 g). ESI-MS (m/z): 547.2 [M+H] ⁺ .

Step 3: ((((R)-1-(6-amino-9H-indol-9-yl)propan-2-yl)oxy)methyl)-4-(piperidin-4-yl)-benzene Synthesis of base-monophosphate (compounds 1-4)

Compound 1-3 (200 mg, 0.4 mmol) was added to dichloromethane (5.0 mL) at room temperature. After the compound was dissolved, trifluoroacetic acid (0.2 mL) was added, and the mixture was allowed to react at room temperature for 2.5 h. The title compound was trifluoroacetate (200 mg). ESI-MS (m/z): 447.2 [M+H] ⁺ .

Step 4: (4R)-6-((4-(4-(((((())))))))) (hydroxy)phosphonyloxy)phenyl)piperidin-1-yl)methyl)-4-(2-chloro-4-fluorophenyl)-2-(thiazol-2-yl)-1 Synthesis of 4-Dihydropyrimidine-5-carboxylic acid ethyl ester (Compound 1-5)

Trifluoroacetate salt of compound 1-4 (163 mg, 0.4 mmol) was dissolved in 1,2-dichloroethane (4 mL) at room temperature, followed by (R)-6-(bromomethyl)-4 -(2-Chloro-4-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (204 mg, 0.4 mmol) and N,N-diiso The propyl ethylamine (0.1 mL) was added and the mixture was evaporated. ESI-MS (m/z): 824.2 [M+H] ⁺ .

Step 5: (4R)-6-((4-(4-(((((()))))))))) ((2-methylbenzyl)oxy)phosphonyloxy)phenyl)piperidin-1-yl)methyl)-4-(2-chloro-4-fluorophenyl)-2- Synthesis of Ethyl (thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate (Compound 1)

Compound 1-5 (30 mg, 0.04 mmol), o-methylbenzyl alcohol (9.0 mg, 0.07 mmol), 1H-benzotriazol-1-yloxytripyrrolidinylphosphonium hexafluorophosphate (at room temperature) PyBOP) (38 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (3 mL). After dissolved, N,N-diisopropylethylamine (19 mg, 0.14 mmol) was added dropwise. After reacting for 4 h at rt, the title compound (9 mg).

Its structural representation is as follows:

^{1 H NMR (400MHz, DMSO-} d 6) δ9.78 (s, 1H), 8.13 (s, 1H), 8.05-8.03 (m, 2H), 7.95 (d, J = 3.13Hz, 1H), 7.44- 7.40 (m, 2H), 7.30-7.14 (m, 9H), 7.03 (d, J = 8.20 Hz, 1H), 6.96 (d, J = 8.12 Hz, 1H), 6.06 (s, 1H), 5.15-5.08 (m, 2H), 4.28-4.16 (m, 2H), 4.11-3.87 (m, 7H), 3.06 (d, J = 10.39 Hz, 1H), 2.88 (d, J = 10.29 Hz, 1H), 2.60- 2.57(m,1H),2.44-2.39(m,1H),2.32-2.24(m,4H),1.88-1.67(m,4H),1.09-1.04(m,6H).ESI-MS(m/z ): 928.2 [M+H] ⁺ .

Example 2(4R)-6-((4-(4-(((((()))))))))) ((S)-1-ethoxy-1-oxopropan-2-yl)oxy)phosphonyl)oxy)phenyl)piperidin-1-yl)methyl)-4-( Ethyl 2-chloro-4-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate (Compound 2)

Compound 1-5 (30 mg, 0.04 mmol), L-ethyl lactate (8.0 mg, 0.07 mmol) and PyBOP (38 mg, 0.07 mmol) were dissolved in N,N-dimethylformamide (3 mL). After the dissolution was completed, N,N-diisopropylethylamine (19 mg, 0.14 mmol) was added dropwise.

Its structural representation is as follows:

^{1 H NMR (400MHz, DMSO-} d 6) δ9.77 (s, 1H), 8.13 (s, 1H), 8.05 (s, 1H), 8.03 (d, J = 23.26Hz, 1H), 7.95 (d, J=3.13Hz,1H),7.44-7.40(m,2H), 7.27-7.16(m,5H),7.05-7.03(m,2H),6.06(s,1H),5.00-4.92(m,1H) , 4.31-3.87 (m, 11H), 3.06 (d, J = 9.75 Hz, 1H), 2.88 (d, J = 11.02 Hz, 1H), 2.60-2.57 (m, 1H), 2.43-2.40 (m, 1H) ), 2.33 - 2.26 (m, 1H), 1.88-1.68 (m, 4H), 1.35 (dd, J = 21.76 Hz, 6.84 Hz, 3H), 1.10 (d, J = 6.22 Hz, 3H), 1.05 (t , J = 7.11 Hz, 3H), 1.17 (dt, J = 7.11 Hz, 3.05 Hz, 3H). ESI-MS (m/z): 924.2 [M+H] ⁺ .

Example 3(4R)-6-((4-(4-(((((()))))))))) (((R)-1-ethoxy-1-oxopropan-2-yl)oxy)phosphonyl)oxy)phenyl)piperidin-1-yl)methyl)-4-( Ethyl 2-chloro-4-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate (Compound 3)

Compound 1-5 (30 mg, 0.04 mmol), D-ethyl lactate (8.0 mg, 0.07 mmol) and PyBOP (38 mg, 0.07 mmol) were dissolved in N,N-dimethylformamide (3 mL). After the dissolution was completed, N,N-diisopropylethylamine (19 mg, 0.14 mmol) was added dropwise.

Its structural representation is as follows:

^{_{1 H NMR (400MHz, CDCl 3}} ) δ9.80 (brs, 1H), 8.34 (s, 1H), 7.99-7.89 (m, 2H), 7.44 (s, 1H), 7.33 (s, 1H), 7.19 ( d, J=8.15 Hz, 2H), 7.14-7.11 (m, 1H), 7.01-6.92 (m, 3H), 6.22 (s, 1H), 5.64 (brs, 2H), 5.05-4.98 (m, 1H) , 4.39-4.34 (m, 1H), 4.23 - 3.79 (m, 10H), 3.02 (d, J = 46.30 Hz, 2H), 2.54-2.38 (m, 3H), 1.90 - 1.86 (m, 4H), 1.51 (dd, J=25.54 Hz, 6.92 Hz, 3H), 1.29-1.21 (m, 6H), 1.14 (t, J = 7.10 Hz, 3H). ESI-MS (m/z): 924.2 [M+H] ⁺ .

Example 4(4R)-6-((4-(4-((2,5,8,11,14,17,20-heptaoxadocosin-22-yloxy)) ((( (R)-1-(6-Amino-9H-indol-9-yl)propan-2-yl)oxy)methyl)phosphonyloxy)phenyl)piperidin-1-yl)methyl) Ethyl 4-(2-chloro-4-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate (Compound 4)

Compound 1-5 (30 mg, 0.04 mmol), heptaethylene glycol monomethyl ether (24.0 mg, 0.07 mmol) and PyBOP (38 mg, 0.07 mmol) were dissolved in N,N-dimethylformamide (3 mL) After the dissolution was completed, N,N-diisopropylethylamine (19 mg, 0.14 mmol) was added dropwise.

Its structural representation is as follows:

^{_{1 H NMR (400MHz, CDCl 3}} ) δ9.78 (brs, 1H), 8.33 (d, J = 2.16Hz, 1H), 7.97 (d, J = 13.35Hz, 1H), 7.90 (dd, J = 5.33Hz , 3.17 Hz, 1H), 7.44 (d, J = 2.98 Hz, 1H), 7.32 (dd, J = 8.45 Hz, 6.24 Hz, 1H), 7.22 - 7.16 (m, 2H), 7.14 - 7.09 (m, 2H) ), 7.01 (d, J = 7.95 Hz, 1H), 6.91 (td, J = 8.36 Hz, 2.27 Hz, 1H), 6.22 (s, 1H), 5.79 (brs, 2H), 4.36 (ddd, J = 14.48) Hz, 6.43 Hz, 2.82 Hz, 1H), 4.30-4.21 (m, 1H), 4.17-4.12 (m, 1H), 4.06-3.98 (m, 4H), 3.88 (d, J = 17.09 Hz, 1H), 3.81-3.73 (m, 1H), 3.64-3.53 (m, 28H), 3.37 (s, 3H), 3.05 (d, J = 10.76 Hz, 1H), 2.92 (d, J = 10.87 Hz, 1H), 2.53 -2.47 (m, 2H), 2.35 (t, J = 8.41 Hz, 1H), 1.89-1.77 (m, 4H), 1.24 (dd, J = 6.02 Hz, 4.16 Hz, 3H), 1.13 (t, J = 7.10 Hz, 3H). ESI-MS (m/z): 573.8 [M / 2+H] ⁺ .

Example 5(4R)-6-((4-(4-(((((())))))))))) (pentyloxy)phosphonyloxy)phenyl)piperidin-1-yl)methyl)-4-(2-chloro-4-fluorophenyl)-2-(thiazol-2-yl) -1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (Compound 5)

Compound 1-5 (30 mg, 0.04 mmol), n-pentanol (6.0 mg, 0.07 mmol) and PyBOP (38 mg, 0.07 mmol) were dissolved in N,N-dimethylformamide (3 mL) at room temperature to dissolve After completion, N,N-diisopropylethylamine (19 mg, 0.14 mmol) was added dropwise.

Its structural representation is as follows:

^{1 H NMR (400MHz, DMSO-} d 6) δ9.77 (s, 1H), 8.13 (s, 1H), 8.04-8.03 (m, 2H), 7.95 (d, J = 3.08Hz, 1H), 7.44- 7.40 (m, 2H), 7.27-7.16 (m, 5H), 7.07 (d, J = 7.99 Hz, 1H), 7.00 (d, J = 8.27 Hz, 1H), 6.06 (s, 1H), 4.30-4.16 (m, 2H), 4.03-3.91 (m, 9H), 3.06 (d, J = 10.38 Hz, 1H), 2.88 (d, J = 11.44 Hz, 1H), 2.60-2.58 (m, 1H), 2.44- 2.39(m,1H),2.33-2.26(m,1H),1.87-1.67(m,4H),1.53-1.43(m,2H),1.23-1.18(m,4H),1.11(t,J=6.57 Hz, 3H), 1.05 (t, J = 7.11 Hz, 3H). ESI-MS (m/z): 894.3 [M+H] ⁺ .

Example 6(4R)-6-((4-(4-(((((())))))))))) ((p-Pentyloxy)methoxy)phosphonyl)oxy)phenyl)piperidin-1-yl)methyl)-4-(2-chloro-4-fluorophenyl)-2- Ethyl (thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate (Compound 6)

Compound 1-5 (30 mg, 0.04 mmol) was dissolved in N-methylpyrrolidone (2 mL) at room temperature, and after complete dissolution, N,N-diisopropylethylamine (9 mg, 0.07 mmol) and Iomethyl valerate (13 mg, 0.06 mmol) was added.

Its structural representation is as follows:

^{_{1 H NMR (400MHz, CDCl 3}} ) δ9.19 (s, 2H), 8.38-8.34 (m, 1H), 7.95-7.89 (m, 2H), 7.44 (m, 1H), 7.35-7.30 (m, 1H ), 7.23-7.19 (m, 2H), 7.17-7.08 (m, 2H), 7.03-6.90 (m, 3H), 6.22 (s, 1H), 5.75-5.59 (m, 2H), 4.40-4.34 (m , 2H), 4.11-3.98 (m, 5H), 3.82-2.75 (m, 1H), 3.48-3.35 (m, 2H), 3.11-3.01 (m, 1H), 2.94-2.84 (m, 1H), 2.59 - 2.42 (m, 2H), 1.98-1.84 (m, 4H), 1.34-1.12 (m, 15H). ESI-MS (m/z): 938.2 [M+H] ⁺ .

Example 7(4R)-6-((4-(((((((()))))))))))))) ((S)-1-ethoxy-1-oxopropan-2-yl)oxy)phosphonyl)oxy)-3,3-difluoropiperidin-1-yl)methyl)-4- (2-Chloro-4-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (Compound 27)

Step 1: 4-(((((()))))))))))))))))) Synthesis of tert-butyl-3,3-difluoropiperidine-1-carboxylate (compound 7-2)

(R)-9-(2-methoxymethoxypropyl)-adenine (55 mg, 0.19 mmol) and tert-butyl 3,3-difluoro-4-hydroxypiperidine-1-carboxylate at room temperature (Compound 7-1) (30 mg, 0.13 mmol) was dissolved in pyridine (2 mL), and dicyclohexylcarbodiimide (80 mg, 0.39 mmol) was added under a nitrogen atmosphere, and the mixture was warmed to 80 ° C overnight. The title compound (10 mg) was obtained after purification. ESI-MS (m/z): 507.1 [M+H] ⁺ .

Step 2: ((((R)-1-(6-Amino-9H-indol-9-yl)propan-2-yl)oxy)methyl)-(3,3-difluoropiperidin-4- Synthesis of mono)triphosphate trifluoroacetate (compound 7-3)

Compound 7-2 (10 mg, 0.02 mmol) was added to dichloromethane (1.0 mL) at room temperature, and trifluoroacetic acid (0.2 mL) was added and dissolved. The reaction mixture was evaporated to drynessjjjjjjjjjj ESI-MS (m/z): 407.1 [M+H] ⁺ .

Step 3: (4R)-6-((4-(((((((())))))))))) Hydroxy)phosphono)oxy)-3,3-difluoropiperidin-1-yl)methyl)-4-(2-chloro-4-fluorophenyl)-2-(thiazol-2-yl)- Synthesis of Ethyl 1,4-dihydropyrimidine-5-carboxylate (Compound 7-4)

Compound 7-3 (10 mg, 0.025 mmol) was dissolved in 1,2-dichloroethane (2 mL) at room temperature, followed by (R)-6-(bromomethyl)-4-(2-chloro- Ethyl 4-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate (14 mg, 0.030 mmol) and N,N-diisopropylethylamine (0.1 (mL), after the addition was completed, the reaction was allowed to proceed overnight at room temperature. The title compound (10 mg) was obtained after purification. ESI-MS (m/z): 784.1 [M+H] ⁺ .

Step 4: (4R)-6-((4-((((((((())))))))))) ((S)-1-ethoxy-1-oxopropan-2-yl)oxy)phosphonyl)oxy)-3,3-difluoropiperidin-1-yl)methyl)-4- Synthesis of (2-Chloro-4-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (Compound 27)

Compound 7-4 (40 mg, 0.05 mmol), L-ethyl lactate (10 mg, 0.1 mmol) and PyBOP (52 mg, 0.1 mmol) were dissolved in N,N-dimethylformamide (3 mL). After the dissolution was completed, N,N-diisopropylethylamine (26 mg, 0.2 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. The title compound (10 mg) was obtained after purification.

Its structural representation is as follows:

1H NMR (400MHz, CDCl ₃ ) δ 9.46 (d, J = 11.90 Hz, 1H), 8.35 (t, J = 2.16 Hz, 1H), 8.07-7.98 (m, 1H), 7.85 (d, J = 3.24) , 1H), 7.43 (d, J = 2.87 Hz, 1H), 7.31 (ddd, J = 9.02, 6.15, 3.19 Hz, 1H), 7.13 (dd, J = 8.60, 2.57 Hz, 1H), 6.93 (td, J=8.32, 2.42 Hz, 1H), 6.21 (t, J=2.29 Hz, 1H), 5.91 (s, 2H), 5.02-4.90 (m, 1H), 4.78-4.67 (m, 1H), 4.45-4.34 (m, 1H), 4.27-3.90 (m, 9H), 3.74-3.66 (m, 1H), 3.02-2.63 (m, 3H), 2.25-1.99 (m, 3H), 1.55-1.44 (m, 3H) , 1.31-1.23 (m, 6H), 1.13 (t,J = 7.16, 1.39 Hz, 3H). ESI-MS (m/z): 884.1 [M+H] ⁺ .

Example VIII (4R)-6-((4-(((((((()))))))))))) Hydroxy)phosphono)oxy)-3,3-difluoropyrrolidin-1-yl)methyl)-4-(2-chloro-4-fluorophenyl)-2-(thiazol-2-yl)- Ethyl 1,4-dihydropyrimidine-5-carboxylate (Compound 28)

Step 1: 4-(((((()))))))))))))))))) Synthesis of -3,3-difluoropyrrolidine-1-carboxylic acid tert-butyl ester (Compound 8-2)

Tenofovir (Compound 8-1) (966 mg, 3.36 mmol) and 3,3-difluoro-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester (500 mg, 2.24 mmol) were dissolved in pyridine at room temperature. (6 mL), dicyclohexylcarbodiimide (1.39 g, 6.72 mmol) was added under a nitrogen atmosphere, and the mixture was heated to 80 ° C overnight. The title compound (240 mg) was obtained after purification. ESI-MS (m/z): 493.1 [M+H] ⁺ .

Step 2: ((((R)-1-(6-Amino-9H-indol-9-yl)propan-2-yl)oxy)methyl)-4,4-difluoropyrrolidin-3-yl Synthesis of monophosphate trifluoroacetate (compound 8-3)

Compound 8-2 (100 mg, 0.2 mmol) was added to dichloromethane (1.5 mL) at room temperature. After the mixture was dissolved, trifluoroacetic acid (0.5 mL) was added, and the mixture was stirred at room temperature. The solvent was evaporated to dryness crystall ESI-MS (m/z): 393.1 [M+H] ⁺ .

Step 3: (4R)-6-((4-(((((((())))))))))) Hydroxy)phosphono)oxy)-3,3-difluoropyrrolidin-1-yl)methyl)-4-(2-chloro-4-fluorophenyl)-2-(thiazol-2-yl)- Synthesis of Ethyl 1,4-dihydropyrimidine-5-carboxylate (Compound 28)

Compound 8-3 (80 mg, 0.2 mmol) was dissolved in dichloromethane (4 mL) at room temperature, then (R)-6-(bromomethyl)-4-(2-chloro-4-fluorophenyl) Ethyl 2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate (100 mg, 0.22 mmol) and N,N-diisopropylethylamine (0.5 mL) Reactive overnight at room temperature. The title compound (80 mg) was obtained after purification.

Its structural representation is as follows:

¹ H NMR (400 MHz, methanol-d ₄ ) δ 8.31 (d, J = 1.6 Hz, 1H), 8.18 (d, J = 2.7 Hz, 1H), 7.78 (dd, J = 5.3, 3.2 Hz, 1H) , 7.67 (dd, J = 5.8, 3.1 Hz, 1H), 7.46-7.39 (m, 1H), 7.21 (dd, J = 8.8, 2.6 Hz, 1H), 7.08-7.02 (m, 2H), 6.14 (d) , J=2.0 Hz, 1H), 4.87-4.75 (m, 1H), 4.38-4.32 (m, 1H), 4.24-4.00 (m, 5H), 3.88-3.84 (m, 1H), 3.77-3.68 (m , 1H), 3.40-3.36 (m, 1H), 3.29-2.95 (m, 2H), 2.78-2.76 (m, 1H), 1.13-1.07 (m, 6H). ESI-MS (m/z): 770.1 [M+H] ⁺ .

Example 9 (4R)-6-((4-(((((((()))))))))))) ((S)-1-ethoxy-1-oxopropan-2-yl)oxy)phosphonyl)oxy)-3,3-difluoropyrrolidin-1-yl)methyl)-4- (2-Chloro-4-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (Compound 29)

The title compound (20 mg) was obtained from the compound (m. ESI-MS (m/z): 870.1 [M+H] ⁺ .

Example X(4R)-6-((4-((((((()))))))))))) ((R)-1-ethoxy-1-oxopropan-2-yl)oxy)phosphonyl)oxy)-3,3-difluoropyrrolidin-1-yl)methyl)-4- (2-Chloro-4-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (Compound 30)

The title compound (10 mg) was obtained from the title compound (d. ESI-MS (m/z): 870.1 [M+H] ⁺ .

Example 11 2-(((((())))))))) - 6-(2-chloro-4-fluorophenyl)-5-(ethoxycarbonyl)-2-(thiazol-2-yl)-3,6-dihydropyrimidin-4-yl)methyl)- 4,4-difluoropyrrolidin-3-yl)oxy)phosphono)oxy)acetic acid (Compound 31)

Compound 28 (30 mg, 0.04 mmol) was dissolved in N-methylpyrrolidone (2 mL), bromoacetic acid (27 mg, 0.20 mmol) and potassium carbonate (16 mg, 0.12 mmol). °C reaction for 2h. The reaction mixture was filtered. ESI-MS (m/z): 828.0 [M+H] ⁺ .

EXAMPLE Twelve (4R)-4-(2-chloro-4-fluorophenyl)-6-((3,3-difluoro-4-((((())))) -5-(5-Methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl)methoxy)(((S)-1- Isopropoxy-1-oxopropan-2-yl)amino)phosphono)oxy)pyrrolidin-1-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine -5-carboxylate (Compound 32)

Step 1: (4R)-4-(2-Chloro-4-fluorophenyl)-6-((4-((dichlorophosphonyl)oxy)-3,3-difluoropyrrolidin-1-yl) Synthesis of ethyl (ethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate (Compound 12-2)

(4R)-4-(2-chloro-4-fluorophenyl)-6-((3,3-difluoro-4-hydroxypyrrolidin-1-yl)methyl)-2-() at room temperature Ethyl thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate (Compound 12-1) (50 mg, 0.1 mmol) was dissolved in dichloromethane (2 mL). At ° C, phosphorus oxychloride (0.2 mL) was added dropwise, and triethylamine (0.2 mL) was added dropwise, and the mixture was reacted at -20 ° C for 30 min. The reaction solution was directly dried to dryness, and the obtained crude material was applied to the next reaction without purification.

Step 2: (4R)-4-(2-chloro-4-fluorophenyl)-6-((3,3-difluoro-4-(((()))) Oxopropan-2-yl)amino)(pentafluorophenoxy)phosphonyloxy)pyrrolidin-1-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydro Synthesis of pyrimidine-5-carboxylate ethyl ester (compound 12-3)

Compound 12-2 (60 mg, 0.1 mmol) was dissolved in dichloromethane (2 mL) at room temperature, and the temperature was lowered to -20 ° C under nitrogen, and L-alanine isopropyl ester hydrochloride (50 mg, 0.3) was added dropwise. A solution of methylene chloride (1.0 mL) was added, and a solution of triethylamine (0.2 mL) in dichloromethane (0.5 mL) was evaporated. Under nitrogen protection, the temperature was further lowered to -20 ° C, and a solution of pentafluorophenol (50 mg, 0.3 mmol) and triethylamine (0.1 mL) in dichloromethane (2.0 mL) was added dropwise, and the reaction was carried out at -20 ° C for 1 h. The reaction solution was directly sparged, dissolved in tetrahydrofuran (4.0 mL), and the insoluble material was filtered, and the filtrate was directly used for the next reaction.

Step 3: (4R)-4-(2-chloro-4-fluorophenyl)-6-((3,3-difluoro-4-((((())))) 5-(5-Methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl)methoxy)(((S)-1-iso) Propoxy-1-oxopropan-2-yl)amino)phosphono)oxy)pyrrolidin-1-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine- Synthesis of ethyl 5-carboxylate (Compound 32)

At room temperature, telbivudine (50 mg, 0.2 mmol) was dissolved in tetrahydrofuran (2 mL), the temperature was lowered to -20 ° C under nitrogen, and t-butyl magnesium chloride (0.4 mL, 1 M) was added dropwise. After 1 h, the temperature was further lowered to -20 ° C under nitrogen atmosphere, and the tetrahydrofuran solution obtained in the second step was added dropwise, and the reaction was carried out overnight under nitrogen atmosphere at room temperature. The title compound (2.0 mg) was obtained after purification. ESI-MS (m/z): 918.0 [M+H] ⁺ .

Example XIII(4R)-6-((4-(((((())))))))))))))) (((R)-1-ethoxy-1-oxopropan-2-yl)oxy)phosphonyl)oxy)-3,3-difluoropiperidin-1-yl)methyl)-4 -(2-Chloro-4-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (Compound 33)

The title compound (15 mg) was obtained from m. ESI-MS (m/z): 884.1 [M+H] ⁺ .

Other compounds in the present application can be synthesized by referring to methods similar to those in the above examples.

Biological activity test

The inhibitory effect of the compounds of the invention on hepatitis B virus (HBV) was tested. The cytotoxicity of the compounds of the invention and the effect on viral (HBV) nucleic acid (DNA) replication levels were tested at the virus-cell level.

experiment method

HepG2 2.2.15 cells in logarithmic growth phase were seeded in 96-well plates at a cell concentration of 40/μL. Incubate for 3 days at 37 ° C in a 5% CO ₂ incubator; replace with fresh medium (200 μL/well) before adding the compound. The mother liquor concentration of each test compound was 200 μM. The highest concentration was 200 μM, diluted to a number of different concentrations in DMSO, and 1 μL of the test compound was placed in the corresponding medium well. The final test concentrations of the compounds were 0.06, 0.24, 0.98, 3.9, 15.6, 62.5, 250, 1000 nM (using Calculate the half effective concentration (EC ₅₀ )).

The mother liquor of each test compound was diluted to 600 μM and 60 μM with DMSO, and 1 μL of each was added to the corresponding medium well, and the final test concentrations of the compounds were 3 μM and 0.3 μM (for calculating the percent inhibition).

Blank control: 1 μL of DMSO was added to the corresponding medium wells as a control.

After the test compound was added, it was co-cultured for 10 days at 37 ° C in a 5% CO ₂ incubator, the medium was changed every two days, and the compound was re-added.

150 μL of cell supernatant was collected per well on day 11 for qPCR detection.

The results of the EC _{50 of} the tested compounds and the inhibition rates at the concentrations of 3 μM and 0.3 μM are shown in Tables 1 and 2.

Table 1

化合物Compound	EC₅₀(nM)EC ₅₀ (nM)
22	59.4359.43
33	54.6554.65
2727	52.152.1
2929	5.185.18
3030	8.218.21
3232	26.9526.95
3333	53.153.1

As can be seen from the data in Table 1, the tested compounds showed strong inhibitory activity in the activity test for inhibiting the replication of deoxyribonucleic acid (DNA) of hepatitis B virus (HBV), and some compounds had an EC _{50 of} <60 nM, preferably a compound (for example). The EC _{50 of the} compounds 29 and 30) was <10 nM, indicating that the compound of the present invention has very excellent inhibitory activity.

Table 2

化合物(3μM)Compound (3 μM)	抑制率％Inhibition rate%
11	96.5496.54
22	100.20100.20
33	101.67101.67

From the above results, it was found that the tested compound exhibited an excellent inhibitory effect in the activity test for inhibiting the deoxyribonucleic acid (DNA) replication of hepatitis B virus (HBV) at a concentration of 3 μM. The compound tested also exhibited an excellent inhibitory effect at a concentration of 0.3 μM, indicating that the compound of the present invention has a very excellent inhibitory effect.

Cytotoxicity test

The test compound was diluted to 30 mM with DMSO, diluted to a maximum concentration of 30 mM, and diluted to a plurality of different concentrations. 0.2 μL of each compound was added to a 384-well plate, and 2000/50 μL of HepG2 was added to each well. For 15 cells, the highest concentration of the test compound was 150 μM; 1 μL of DMSO was added to the corresponding wells as a control.

The cells were co-cultured for 4 days at 37 ° C in a 5% CO ₂ incubator.

After 4 days, 50 μL of CellTiter-Glo was added to each well to perform a plate reading test, and a half-cytotoxic concentration (CC ₅₀ ) value was calculated.

Preferred compounds (e.g. compound of Example 5) CC ₅₀ value of greater than 150 M, indicating its low cytotoxicity and high safety.

Various modifications of the invention in addition to those described herein will be apparent to those skilled in the art. Such modifications are also intended to fall within the scope of the appended claims. Each of the references (including all patents, patent applications, journal articles, books, and any other publications) cited in this application are hereby incorporated by reference in their entirety.

Claims

a compound, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein the compound has the structure of formula (I):

among them:

B is selected from:

A is selected from C 1-6 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, optionally substituted by one or more R b , said alkylene, alkenylene or sub An alkynyl group is optionally interrupted by one or more -O-, -NR- or -S-;

Or A is selected from the following groups:

among them
Represents a single bond or a double bond, and is connected to B at the 1 position and to the phosphorus atom (P) at the 2 position;

X, Y and Z are each independently selected from CH 2 , O, S and NR at each occurrence;

R a and R b are each independently selected from the group consisting of halogen, -OH, -CN, -NO 2 , -N(R) 2 , -N 3 , C 1-6 alkyl and C 3-6 ring at each occurrence. alkyl;

R 2 is selected from the group consisting of H, C 1-12 alkyl, C 3-6 cycloalkyl, C 6-14 aryl, 5-14 membered heteroaryl, C 6-20 aralkyl, -C 1-6 Alkyl-COOH, -C 1-6 alkylene-C(=O)OC 1-6 alkyl, -C 1-6 alkylene-OC(=O)-C 1-6 alkyl, -C 1-6 alkylene-OC(=O)OC 1-6 alkyl and -((CH 2 ) i O) m -(CH 2 ) q -OC 1-6 alkyl, each of the above groups optionally One or more selected from the group consisting of halogen, -OH, -CN, -NO 2 , -N(R) 2 , C 1-6 alkyl, halogenated C 1-6 alkyl, C 1-6 alkylthio Substituted with a substituent of a C 3-6 cycloalkyl group;

i and q are each independently 1, 2, 3, 4, 5 or 6 at each occurrence;

m is any integer selected from 0-50, preferably 0-20, particularly preferably 0-6;

R 1 is selected from:

Representing a 3 to 14 membered nitrogen heterocycle, optionally additionally containing 1, 2 or 3 ring members independently selected from N, O, C=O, S, S=O and S(=O) 2 ;

Ar 1 and Ar 2 are each independently selected from a C 6-14 aryl group and a 5-14 membered heteroaryl group, which are optionally selected from one or more selected from the group consisting of halogen, -OH, -CN, -NO 2 , -N Substituent substitution of (R) 2 , C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylthio and C 3-6 cycloalkyl;

L is absent or selected from -O-, -S-, and -NR-;

R 3 and R 4 are each independently selected from the group consisting of H, C 1-4 alkyl and C 3-6 cycloalkyl;

R 5 is attached to the remainder of the molecule with a single or double bond at each occurrence;

R 5 and R 6 are each independently selected from the group consisting of halogen, -OH, -COOH, -CN, -NO 2 , -N(R) 2 , C 1-6 alkyl, halogenated C 1-6 Alkyl, -WC 1-6 alkyl, -C 1-6 alkylene-WR, -WC 1-6 alkylene-W'-R, -WC 2-6 alkenyl, -C 2-6 Alkenyl-WR, -WC 2-6 alkenylene-W'-R and C 3-6 cycloalkyl, wherein the alkylene and alkenylene are optionally further separated by one or more W;

W and W' are each independently selected from the group consisting of O, C(=O), C(=O)O, NR, S, S=O, and S(=O) 2 ;

R is each independently selected from the group consisting of H, C 1-6 alkyl and C 3-6 cycloalkyl;

n each independently is 0, 1, 2, 3, 4 or 5, with the proviso that n is not greater than the number of positions on the corresponding group that can be substituted;

When n is greater than 1, each R a may be the same or different, and each R b may be the same or different, and each R 5 may be the same or different, and each R 6 may be the same or different.
A compound according to claim 1 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein R 2 is selected from the group consisting of H and C 1-12 alkyl, C 3-6 cycloalkyl, C 6-14 aryl, 5-14 membered heteroaryl, C 6-20 aralkyl, -C 1-6 alkylene-C (=O OC 1-6 alkyl, -C 1-6 alkylene-OC(=O)-C 1-6 alkyl, -C 1-6 alkylene-OC(=O)OC 1-6 alkyl And -((CH 2 ) i O) m -(CH 2 ) q -OC 1-6 alkyl, each of the above groups optionally being selected from one or more selected from the group consisting of halogen, -OH, -CN, -NO 2 Substituted with a substituent of -N(R) 2 , C 1-6 alkyl, halo C 1-6 alkyl, C 1-6 alkylthio and C 3-6 cycloalkyl.
A compound according to claim 1 or 2, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein B is selected from the group consisting of:
A compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein A From:

One of the positions is connected to B, and the second position is connected to the phosphorus atom (P).
A compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein R 1 is selected from:
A compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein Ar 1 is selected from:

Wherein R c is each independently selected from the group consisting of F, Cl, Br, I, C 1-6 alkyl, halo C 1-6 alkyl, and C 3-6 cycloalkyl;

Preferably, R c is each independently selected from the group consisting of F, Cl, Br, I, C 1-6 alkyl and C 3-6 cycloalkyl;

Ar 1 is preferably selected from:

Ar 1 is particularly preferred from:
A compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein Ar 2 is selected from:

The above groups are optionally substituted by one or more groups selected from the group consisting of halogen, C 1-6 alkyl, halo C 1-6 alkyl and C 3-6 cycloalkyl;

Preferably, Ar 2 is selected from the group consisting of

The above groups are optionally substituted by one or more groups selected from the group consisting of halogen, C 1-6 alkyl and C 3-6 cycloalkyl.
A compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein R 2 is selected from the group consisting of: C 1-6 alkyl, -((CH 2 ) i O) m -(CH 2 ) q -OC 1-6 alkyl,

among them:

i and q are each independently 1, 2, 3, 4, 5 or 6 at each occurrence;

m is any integer selected from 0-50, preferably 0-20, particularly preferably 0-6;

R 7 , R 8 and R 9 are each independently selected from the group consisting of H, C 1-10 alkyl, C 3-6 cycloalkyl, C 6-20 aryl and C 7-20 aralkyl, said alkyl group, The cycloalkyl, aryl and aralkyl groups are each optionally substituted by one or more substituents selected from the group consisting of halogen, -OH, -CN and -NO 2 ;

Or R 7 and R 8 together with the carbon atom to which they are attached form a C 3-6 cycloalkyl group;

R 10 and R 11 are each independently selected from the group consisting of H, halogen, -OH, -CN, -NO 2 , C 1-10 alkyl, and C 3-6 cycloalkyl.
A compound according to any one of claims 1-8, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein The compound has the structure of formula (II) or formula (II)-1:
A compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein The compound has any structure of the following formula:
A compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, wherein Said compound is selected from:
A pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph thereof, A solvate, metabolite or prodrug and one or more pharmaceutically acceptable carriers, preferably a solid formulation, a liquid formulation or a transdermal formulation.
A method of preparing a pharmaceutical composition, which comprises a compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph thereof, The solvate, metabolite or prodrug is combined with one or more pharmaceutically acceptable carriers.
A compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, metabolite or prodrug thereof, or claim Use of a pharmaceutical composition of 12 for the manufacture of a medicament for the prevention or treatment of a viral disease.
The use according to claim 14, wherein the drug is a drug administered by oral, intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular or transdermal routes.
The use according to claim 14, wherein the viral disease is selected from the group consisting of hepatitis A virus, hepatitis B virus, hepatitis C virus, influenza, herpes and acquired immunodeficiency syndrome (AIDS).
A method of preparing a compound according to any one of claims 1-11, the method comprising the steps of:

or

among them:

R 12 , R 13 and R 14 are each independently selected from the group consisting of F, Cl, Br, I, -NHR, hydroxy, triflate, p-toluenesulfonate and borate;

PG is selected from the group consisting of tert-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, allyloxycarbonyl, trimethylsilyloxycarbonyl, methoxycarbonyl, ethoxycarbonyl, p-toluenesulfonyl, o-nitrobenzenesulfonate Acyl, p-nitrophenylsulfonyl, formyl, acetyl, trifluoroacetyl, propionyl, pivaloyl, phenyl, benzoyl, trityl, benzyl, 2,4-dimethoxy Benzyl and p-methoxybenzyl;

The remaining groups are as defined in any one of claims 1-11;

The first step is carried out in the presence of an organic base or an inorganic base and/or a condensation reagent (particularly preferably DCC, DIC, EDC, BOP, PyAOP or PyBOP) in an aprotic solvent or without solvent;

The second step is carried out under conditions suitable for removal of the PG group;

The third step is carried out in an aprotic solvent in the presence of an organic or inorganic base;

The fourth step is carried out in an aprotic solvent, preferably in the presence of an organic base or an inorganic base and/or a condensation reagent, particularly preferably DCC, DIC, EDC, BOP, PyAOP or PyBOP;

Or the method comprises the steps of:

among them:

R 12 is selected from the group consisting of F, Cl, Br, I, -NHR, hydroxy, triflate, p-toluenesulfonate, and borate;

LG is a leaving group, which is preferably a pentafluorophenyl group and a p-nitrophenyl group;

The remaining groups are as defined in any one of claims 1-11;

The first step is carried out in an aprotic solvent in the presence of an organic or inorganic base;

The second step is carried out in an aprotic solvent in the presence of an organic or inorganic base;

The third step is carried out in an aprotic solvent in the presence of an organic or inorganic base.