CN105315319B - Hepatitis C virus inhibitor and its application - Google Patents

Abstract

The invention belongs to the field of medicinal chemistry, and in particular relates to a class of compounds with good hepatitis C virus inhibitory effect, a preparation method thereof, a composition containing the compound, and the compound or composition as a therapeutic drug for hepatitis C virus infectious disease the use of. The compound of the present invention exhibits excellent antiviral activity, has small cytotoxicity, good safety, and good plasma protein binding rate, is suitable for medicine, and has a very good clinical application prospect.

Description

Hepatitis C virus inhibitor and its application

technical field

The invention belongs to the field of medicinal chemistry, and in particular relates to a class of compounds with good hepatitis C virus inhibitory effect, a preparation method thereof, a composition containing the compound, and the compound or composition as a therapeutic drug for hepatitis C virus infectious disease the use of.

Background technique

Hepatitis C virus (HCV) infection is a prevalent disease worldwide, with more than 200 million chronically infected people worldwide. The infection rate in China is 3.2%, ranking among the top three in the world. The clinical manifestations of HCV infection are diverse, ranging from mild inflammation to severe liver cirrhosis and liver cancer. Chronic hepatitis C can also be complicated by certain extrahepatic manifestations, including rheumatoid arthritis, conjunctivitis sicca, lichen planus, glomerulonephritis, mixed cryoglobulinemia, B-cell lymphoma, and late-onset Skin porphyria, etc., may be caused by abnormal immune response of the body. In the decompensated stage of hepatitis C cirrhosis, various complications can occur, such as ascites and abdominal infection, upper gastrointestinal bleeding, hepatic encephalopathy, hepatorenal syndrome, and liver failure.

HCV belongs to the Flaviviridae family of viruses of the genus Hepatovirus, which is similar in genetic structure to two other genera in the Flaviviridae family, Pestivirus and Flavivirus. Currently, the standard treatments for HCV infection are interferon and combination therapy with interferon and ribavirin. However, only 50% of those treated responded to the approach, and interferon had significant side effects such as influenza-like symptoms, weight loss, and fatigue, while the combination of interferon and ribavirin produced considerable Side effects, including hemolysis, anemia, and fatigue.

In addition, drugs that have been developed for the treatment of HCV infection include protease inhibitors, thiazolidine derivatives, thiazolidine and benzoic anilides, phenanthrenequinones, helicase inhibitors, nucleoside polymerase inhibitors and glioblastoma Toxins, antisense phosphorothioate oligonucleotides, inhibitors of IRES-dependent translation, ribozymes, and nucleoside analogs, etc.

At present, the use of nucleoside phosphate compounds for the treatment of Flaviviridae viruses, especially HCV infection, is an important research and development direction in this field. WO 2006/065335 discloses a fluorinated pyrrolo[2,3,d]pyrimidine nucleoside compound that inhibits HCV virus. US 2006/0241064 discloses nucleoside compounds for the treatment of viral infections caused by viruses of the Flaviviridae family, such as HCV. WO 2008/121634 discloses nucleoside phosphoramidate compounds for use in the treatment of viral infections in mammals.

Despite the above disclosures, there is still a great need for effective compounds for the treatment and/or prevention of HCV infection.

SUMMARY OF THE INVENTION

One object of the present invention is to provide a compound of general formula I or its stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal for treating and/or preventing hepatitis C virus infection:

Another object of the present invention is to provide a process for preparing the compounds of general formula I of the present invention or stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or crystals thereof.

A further object of the present invention is to provide a composition comprising a compound of general formula I or a stereoisomer, a pharmaceutically acceptable salt, hydrate, solvate or crystalline and a pharmaceutically acceptable carrier of the present invention and a composition comprising The composition of the compound of general formula I of the present invention or its stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal and another antiviral drug.

Still another object of the present invention is to provide a compound of general formula I or a stereoisomer, a pharmaceutically acceptable salt, hydrate, solvate or crystal of the present invention for the treatment and/or prevention of hepatitis C virus infection and Use of the compound of general formula I or its stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal of the present invention in the preparation of a medicament for the treatment and/or prevention of viral infection.

For the above-mentioned purpose of the invention, the present invention provides the following technical solutions:

In the first aspect, the present invention provides the compound represented by the general formula I or its stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal:

in:

R ₁ is selected from H, alkyl and haloalkyl;

R _{is selected from H and halogen;}

_R is selected from H, OH and alkoxy;

_R4 is selected from H and alkyl, wherein said alkyl is optionally selected from alkyl, cycloalkyl, heterocycloalkyl, alkoxy, alkylamino, halo, hydroxy, amino, nitro, cyano , alkyl acyl, amino acyl, alkylamino acyl, sulfonyl, sulfinyl, mercapto, aryl and heteroaryl group substituted with one or more groups;

R _is selected from H, alkyl, cycloalkyl, aryl and heteroaryl, wherein said alkyl, cycloalkyl, aryl and heteroaryl are optionally selected from alkyl, cycloalkyl, heterocycle Among alkyl, alkoxy, alkylamino, halogen, hydroxyl, amino, nitro, cyano, alkylacyl, aminoacyl, alkylaminoacyl, sulfonyl, sulfinyl, mercapto, aryl and heteroaryl substituted with one or more groups;

Cy ₁ is selected from aryl and heteroaryl, wherein said aryl and heteroaryl are optionally selected from halogen, hydroxyl, alkyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, amino, Substitution of one or more groups of alkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, nitro, and cyano;

Cy ₂ is selected from hydrogen, aryl, heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein said aryl, heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, Hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, amino, alkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, nitro One or more groups in cyano group and cyano group are substituted;

Provided that Cy ₁ is an aryl group, Cy ₂ is not hydrogen or aryl, and the compound of general formula I of the present invention is not the following compounds:

(2S)-2-(((4-(1,2,3-thiadiazol-4-yl)phenyl-1-yl)oxy)(((2R,3R,4R,5R)-5-( 2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino ) isopropyl propionate;

(2S)-2-(((2-(quinoxalin-5-yl)phenyl-1-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl;

(2S)-2-(((4-(thiazol-2-yl)phenyl-1-yl)oxy)(((2R,3R,4R,5R)-5-(2,4-dioxo- Isopropyl 3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate;

(2S)-2-(((4-(thiazol-5-yl)phenyl-1-yl)oxy)(((2R,3R,4R,5R)-5-(2,4-dioxo- Isopropyl 3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate;

(2S)-2-(((1H-Indol-5-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidine Isopropyl 1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate;

(2S)-2-(((4-Fluoro-2-methyl-1H-indol-5-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate ;

(2S)-2-(((3-methoxycarbonylmethyl-benzofuran-7-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo- Isopropyl 3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate;

(2S)-2-(((4-oxo-2-phenyl-4H-benzopyran-6-yl)oxy)(((2R,3R,4R,5R)-5-(2, 4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propane isopropyl acid;

(2S)-2-(((quinoxalin-5-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidine- Isopropyl 1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate;

(2S)-2-(((4-oxo-2-phenylchroman-7-yl)oxy)(((2R,3R,4R,5R)-5-(2,4 -Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionic acid isopropyl ester;

(2S)-2-((((4-(1H-1,2,4-triazol-1-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2, 4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propane isopropyl acid;

(2S)-2-(((4-(pyrimidin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4- Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl;

(2S)-2-(((S)-((4-Fluoro-1,2-dimethyl-1H-indol-5-yl)oxy)(((2R,3R,4R,5R)- 5-(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphorus Acyl)amino)isopropyl propionate;

(2S)-2-((((6-Fluoro-3-methylbenzo[d]isoxazol-5-yl)oxy)(((2R,3R,4R,5R)-5-(2 ,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino) isopropyl propionate;

(2S)-2-((((6-Fluorobenzo[c][1,2,5]thiadiazol-5-yl)oxy)(((2R,3R,4R,5R)-5- (2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl) amino) isopropyl propionate; and

(2S)-2-(((4-(thiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4- Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionic acid isopropyl ester.

In some preferred embodiments, R ₁ is selected from H, C _1-6 alkyl, haloC _1-6 alkyl, R ₂ is selected from H, fluorine, chlorine, bromine, and R ₃ is selected from H, OH.

In other preferred embodiments, R ₁ is selected from H, C _1-3 alkyl, R ₂ is selected from fluorine, chlorine, and R ₃ is selected from H, OH.

In some preferred embodiments, R ₄ is selected from H, C _1-6 alkyl, and R ₅ is selected from C _1-6 alkyl and C _3-8 cycloalkyl, wherein said alkyl and cycloalkyl optionally selected from alkyl, cycloalkyl, heterocycloalkyl, alkoxy, alkylamino, halo, hydroxy, amino, nitro, cyano, alkylacyl, aminoacyl, alkylaminoacyl, sulfonyl , sulfinyl, mercapto, aryl and heteroaryl are substituted with one or more groups.

In other preferred embodiments, R ₄ is selected from H, C _1-3 alkyl, R ₅ is selected from C _1-6 alkyl and C _3-6 cycloalkyl, wherein said alkyl and cycloalkane The group can be selected from C _1-6 alkyl, C _3-8 cycloalkyl, C _3-8 heterocycloalkyl, C _1-6 alkoxy, C _1-6 alkylamino, halogen, hydroxyl, amino , nitro, cyano, C _1-6 alkyl acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, aryl and heteroaryl substituted with one or more groups .

In other preferred embodiments, R ₄ is selected from H, C _1-3 alkyl, R ₅ is selected from C _1-3 alkyl and C _3-6 cycloalkyl, wherein said alkyl and cycloalkane The group can be selected from C _1-3 alkyl, C _3-6 cycloalkyl, C _3-6 heterocycloalkyl, C _1-3 alkoxy, C _1-3 alkylamino, halogen, hydroxyl, amino , nitro, cyano, C _1-3 alkyl acyl, aminoacyl, C _1-3 alkylamino acyl, sulfonyl, sulfinyl, mercapto, phenyl and heteroaryl substituted with one or more groups .

In some preferred embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein said benzene ring, heteroaryl, heterocycloalkyl and heterocycle Cycloalkenyl is optionally selected from halogen, hydroxy, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, aminoacyl, C _{1 -6} Substituted with one or more groups of alkylaminoacyl, nitro, and cyano.

In other preferred embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from heteroaryl, C _3-8 heterocycloalkyl and C _3-8 heterocycloalkenyl, wherein said benzene ring, heterocyclo Aryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxy, C _1-3 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, Halogenated C _1-3 alkyl, C _1-3 alkoxy, halogenated C _1-3 alkoxy, amino, C _1-3 alkylamino, C _1-3 alkylacylamino, C _1-3 Substituted with one or more groups of alkylacyl, aminoacyl, _C1-3 alkylaminoacyl, nitro, and cyano.

In other preferred embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from heteroaryl, C _3-6 heterocycloalkyl and C _3-6 heterocycloalkenyl, wherein said benzene ring, heterocyclo Aryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxy, C _1-3 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, Halogenated C _1-3 alkyl, C _1-3 alkoxy, halogenated C _1-3 alkoxy, amino, C _1-3 alkylamino, C _1-3 alkylacylamino, C _1-3 Substituted with one or more groups of alkylacyl, aminoacyl, _C1-3 alkylaminoacyl, nitro, and cyano.

In other preferred embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from five- or six-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl containing at least one N atom, wherein the benzene ring, five- or six-membered heteroaryl, heterocycloalkyl or heterocycloalkenyl optionally selected from halogen, hydroxy, _C1-6 alkyl, _C2-4 alkenyl, _C2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkane One or more groups of acylamino, C _1-6 alkyl acyl, aminoacyl, C _1-6 alkyl aminoacyl, nitro, and cyano are substituted.

In other preferred embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from five- or six-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl containing 1-3 N atoms, wherein the The benzene ring, five- or six-membered heteroaryl, heterocycloalkyl or heterocycloalkenyl are optionally selected from halogen, hydroxyl, _C1-6 alkyl, _C2-4 alkenyl, _C2-4 alkyne base, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1- Substituted with one or more groups of ₆ alkylacylamino, _C1-6 alkylacyl, aminoacyl, _C1-6 alkylaminoacyl, nitro, and cyano.

In other preferred embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from five- or six-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl containing only 1 S atom, wherein said Benzene ring, five- or six-membered heteroaryl, heterocycloalkyl or heterocycloalkenyl optionally selected from halogen, hydroxy, _C1-6alkyl , _C2-4alkenyl , _C2-4alkynyl , C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 One or more groups of alkylacylamino, _C1-6 alkylacyl, aminoacyl, _C1-6 alkylaminoacyl, nitro, and cyano are substituted.

In some specific embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from six-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl containing 1 N atom, wherein said benzene ring, heteroaryl group, heterocycloalkyl or heterocycloalkenyl optionally selected from halogen, hydroxy, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogen Substituted C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkane One or more groups of acyl, aminoacyl, C _1-6 alkylaminoacyl, nitro, and cyano are substituted.

In other specific embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from five-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl containing only 1 S atom, wherein the benzene ring, Heteroaryl, heterocycloalkyl or heterocycloalkenyl is optionally selected from halogen, hydroxy, _C1-6 alkyl, _C2-4 alkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl , halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1- Substituted with one or more groups of ₆ alkyl acyl, amino acyl, C _1-6 alkyl amino acyl, nitro, and cyano.

In some other specific embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from five-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl containing 2 N atoms, wherein said benzene ring, heterocyclic Aryl, heterocycloalkyl or heterocycloalkenyl are optionally selected from halogen, hydroxy, _C1-6 alkyl, _C2-4 alkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, Halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkylacylamino, C _1-6 Substituted with one or more groups of alkylacyl, aminoacyl, C _1-6 alkylaminoacyl, nitro, and cyano.

In other specific embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from five-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl containing 1 N atom and 1 sulfur atom, wherein the The benzene ring, heteroaryl, heterocycloalkyl or heterocycloalkenyl is optionally selected from halogen, hydroxyl, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _{3- 6} cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkylacylamino , C _1-6 alkyl acyl group, amino acyl group, C _1-6 alkyl amino acyl group, nitro group, cyano group is substituted with one or more groups.

In other specific embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from five-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl containing 1 N atom and 1 oxygen atom, wherein the The benzene ring, heteroaryl, heterocycloalkyl or heterocycloalkenyl is optionally selected from halogen, hydroxyl, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _{3- 6} cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkylacylamino , C _1-6 alkyl acyl group, amino acyl group, C _1-6 alkyl amino acyl group, nitro group, cyano group is substituted with one or more groups.

In other preferred embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from furan ring, pyrrole ring, thiophene ring, imidazole ring, pyrazole ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole Ring, triazole ring, thiadiazole ring, oxadiazole ring, pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring, benzoazepine ring, benzoxane ring, benzothiane ring, benzene Diazepine ring, benzodioxane ring, benzodithiane ring, benzoxazepine ring, benzothiazepine ring, wherein said furan ring, pyrrole ring, thiophene ring, imidazole ring, Pyrazole ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole ring, triazole ring, thiadiazole ring, oxadiazole ring, pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring, benzene Benzodiazepine, benzoxazepine, benzothiazepine, benzodiazepine, benzodiazepine, benzodiazepine, benzoxazepine, benzothiazepine Ring is optionally selected from halogen, hydroxy, _C1-6 alkyl, _C2-4 alkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, halogenated _C1-6 alkyl, _{C1 -6} alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, aminoacyl, C _1-6 One or more groups of alkylaminoacyl, nitro, and cyano are substituted.

In other preferred embodiments, Cy ₁ is selected from benzene ring, Cy ₂ is selected from furan ring, pyrrole ring, thiophene ring, imidazole ring, pyrazole ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole Ring, triazole ring, thiadiazole ring, oxadiazole ring, pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring, benzoazepine ring, benzoxane ring, benzothiane ring, benzene Diazepine ring, benzodioxane ring, benzodithiane ring, benzoxazepine ring, benzothiazepine ring, wherein said furan ring, pyrrole ring, thiophene ring, imidazole ring, Pyrazole ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole ring, triazole ring, thiadiazole ring, oxadiazole ring, pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring, benzene Benzodiazepine, benzoxazepine, benzothiazepine, benzodiazepine, benzodiazepine, benzodiazepine, benzoxazepine, benzothiazepine Ring is optionally selected from halogen, hydroxy, _C1-6 alkyl, _C2-4 alkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, halogenated _C1-6 alkyl, _{C1 -6} alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, aminoacyl, C _1-6 One or more groups of alkylaminoacyl, nitro, and cyano are substituted, and Cy ₂ is at the para or meta position of Cy ₁ .

In some preferred embodiments, the compound of general formula I according to the present invention, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein Cy ₂ is in the para or meta position of Cy ₁ bit. The inventors of the present invention unexpectedly found that when Cy ₂ in the general formula I is in the para or meta position of Cy ₁ , the compounds of the present invention have very excellent antiviral activity.

In other preferred embodiments, Cy ₁ is selected from naphthalene ring, heteroaryl, Cy ₂ is selected from hydrogen, aryl, heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein said naphthalene ring, Aryl, heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxy, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 Cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkylacylamino, One or more groups of C _1-6 alkyl acyl, aminoacyl, C _1-6 alkyl aminoacyl, nitro, and cyano are substituted.

In other preferred embodiments, Cy ₁ is selected from naphthalene ring, heteroaryl, Cy ₂ is selected from hydrogen, aryl, heteroaryl, C _3-8 heterocycloalkyl and C _3-8 heterocycloalkenyl , wherein the naphthalene ring, aryl, heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxyl, C _1-6 alkyl, C _2-4 alkenyl, C _{2- 4alkynyl} , C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C Substituted with one or more groups of _1-6 alkylacylamino, _C1-6 alkylacyl, aminoacyl, _C1-6 alkylaminoacyl, nitro, and cyano. In other preferred embodiments, Cy ₁ is selected from naphthalene ring, heteroaryl, Cy ₂ is selected from hydrogen, aryl, heteroaryl, C _3-6 heterocycloalkyl and C _3-6 heterocycloalkenyl , wherein the naphthalene ring, aryl, heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxyl, C _1-3 alkyl, C _2-4 alkenyl, C _{2- 4alkynyl} , C _3-6 cycloalkyl, halogenated C _1-3 alkyl, C _1-3 alkoxy, halogenated C _1-3 alkoxy, amino, C _1-3 alkylamino, C Substituted with one or more groups of _1-3 alkylacylamino, _C1-3 alkylacyl, aminoacyl, _C1-3 alkylaminoacyl, nitro, and cyano.

In other preferred embodiments, Cy ₁ is selected from a five- or six-membered heterocyclic ring containing at least one N atom of benzo, wherein the five- or six-membered heterocyclic ring containing at least one N atom of benzo is optionally Selected from halogen, hydroxyl, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Of alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, amino acyl, C _1-6 alkyl amino acyl, nitro, cyano One or more groups are substituted, and Cy ₂ is hydrogen. In other preferred embodiments, Cy ₁ is selected from a five- or six-membered heterocycle containing only one S atom, wherein the five- or six-membered heterocycle containing only one S atom is optionally Selected from halogen, hydroxyl, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Of alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, amino acyl, C _1-6 alkyl amino acyl, nitro, cyano One or more groups are substituted, and Cy ₂ is hydrogen.

In some specific embodiments, Cy ₁ is selected from a benzo five- or six-membered heteroaromatic ring containing 1, 2, or 3 nitrogen atoms, wherein the benzo five- or six-membered contains 1, 2, or 3 The heteroaromatic ring of the nitrogen atom is optionally selected from halogen, hydroxy, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, halogenated C _1-6 alkyl, C _1-6 alkane Oxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, aminoacyl, C _1-6 alkylamino One or more groups of acyl, nitro, and cyano are substituted, and Cy ₂ is hydrogen.

In other specific embodiments, Cy ₁ is selected from a benzoquinta-membered heteroaromatic ring containing 1 or 2 nitrogen atoms, wherein the benzoquinta-membered heteroaromatic ring containing 1 or 2 nitrogen atoms is optionally Selected from halogen, hydroxyl, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Of alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, amino acyl, C _1-6 alkyl amino acyl, nitro, cyano One or more groups are substituted, and Cy ₂ is hydrogen.

In other specific embodiments, Cy ₁ is selected from a benzoquinone-membered heteroaromatic ring containing 1 nitrogen atom and 1 oxygen atom, wherein the benzoquinone contains 1 nitrogen atom and 1 oxygen atom The heteroaromatic ring is optionally selected from halogen, hydroxy, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, Halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, aminoacyl, C _1-6 alkyl aminoacyl, nitro One or more groups of cyano group and cyano group are substituted, and Cy ₂ is hydrogen.

In other specific embodiments, Cy ₁ is selected from a five-membered heteroaromatic ring containing only one S atom, wherein the five-membered heteroaromatic ring containing only one S atom is optionally selected from Halogen, hydroxyl, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy one of _C1-6 alkylamino, C1-6 alkylamino, _C1-6 alkylacylamino, _C1-6 alkylacyl, aminoacyl, _C1-6 alkylaminoacyl, nitro, cyano or Multiple groups are substituted, and Cy ₂ is hydrogen.

In other preferred embodiments, Cy ₁ is selected from furan ring, pyrrole ring, thiophene ring, imidazole ring, pyrazole ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole ring, triazole ring, Thiadiazole ring, oxadiazole ring, pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring, benzazepine ring, benzoxane ring, benzothiane ring, benzodiazepine ring, benzene Dioxane, benzodithia, benzoxazepine, benzothiazepine, Cy ₂ is selected from hydrogen, benzene ring, furan ring, pyrrole ring, thiophene ring, imidazole ring, pyrazole Ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole ring, triazole ring, thiadiazole ring, oxadiazole ring, pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring, benzazepine Heterocycles, benzoxazepines, benzothiazepines, benzodiazepines, benzodioxanes, benzodiazepines, benzoxazepines, benzothiazepines, Wherein the benzene ring, thiophene ring, imidazole ring, pyrazole ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole ring, triazole ring, thiadiazole ring, oxadiazole ring, pyridine ring , pyrimidine ring, pyrazine ring, pyridazine ring, benzazepine ring, benzoxane ring, benzodiazepine ring, benzodiazepine ring, benzodioxane ring, benzodithiane ring , benzoxazepine, benzothiazepine optionally selected from halogen, hydroxy, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, halogenated C _1-6 Alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, aminoacyl , C _1-6 alkylaminoacyl, nitro, cyano group is substituted with one or more groups.

In other preferred embodiments, Cy ₁ is selected from furan ring, pyrrole ring, thiophene ring, imidazole ring, pyrazole ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole ring, triazole ring, Thiadiazole ring, oxadiazole ring, pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring, benzofuran ring, benzopyrrole ring, benzothiophene ring, benzimidazole ring, benzopyrazole ring, benzothiazole ring, benzisothiazole ring, benzoxazole ring, benzisoxazole ring, benzotriazole ring, benzothiadiazole ring, benzoxadiazole ring, benzopyridine ring, Benzopyrimidine ring, benzopyrazine ring, benzopyridazine ring, Cy ₂ is hydrogen, wherein said Cy ₁ is optionally selected from halogen, hydroxyl, C _1-6 alkyl, halogenated C _1-6 Alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, aminoacyl , C _1-6 alkylaminoacyl, nitro, cyano group is substituted with one or more groups.

In some preferred embodiments, the present invention provides a compound of formula Ia or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof:

in:

R _a1 is selected from heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein said heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxy, amino, alkyl, alkene alkynyl, alkynyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, alkylamino, alkylacyl, alkoxyacyl, aminoacyl, alkylaminoacyl, sulfonyl, sulfinyl, mercapto, nitro , one or more groups in the cyano group are substituted;

R _a2 is selected from hydrogen, halogen, hydroxyl, amino, alkyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, alkylamino, alkylacyl, alkoxyacyl, aminoacyl, alkylaminoacyl, sulfonic acid Acyl, sulfinyl, mercapto, nitro and cyano groups; or

R _a1 , R _a2 together with the carbon atoms to which they are attached form a heteroaryl, heterocycloalkyl or heterocycloalkenyl;

R _a3 , R _a4 , R _a5 are independently selected from hydrogen, halogen, hydroxy, amino, alkyl, cycloalkyl, haloalkyl, alkoxy, haloalkoxy, alkylamino, alkylacyl, alkoxyacyl, Aminoacyl, alkylaminoacyl, sulfonyl, sulfinyl, mercapto, nitro, cyano;

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ have the definitions in formula I.

In some embodiments, a compound of general formula Ia or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof according to the present invention, wherein R ₁ is selected from H, C _1-6 alkyl and halogenated C _1-6 alkyl, preferably selected from H and C _1-3 alkyl; R ₂ is selected from H, fluorine, chlorine and bromine, preferably selected from fluorine and chlorine; R ₃ is selected from H and OH; R ₄ is selected from H and C _1-6 alkyl, wherein said alkyl is optionally selected from C _1-6 alkyl, C _3-8 cycloalkyl, C _3-8 heterocycloalkyl, C _1-6 Alkoxy, C _1-6 alkylamino, halogen, hydroxyl, amino, nitro, cyano, C _1-6 alkyl acyl, aminoacyl, C _1-6 alkylaminoacyl, sulfonyl, sulfinyl, One or more groups in mercapto, aryl and heteroaryl are substituted, preferably, R ₄ is selected from H and C _1-3 alkyl, wherein said alkyl is optionally selected from C _1-3 alkyl , C _3-6 cycloalkyl, C _3-6 heterocycloalkyl, C _1-3 alkoxy, C _1-3 alkylamino, halogen, hydroxyl, amino, nitro, cyano, C _1-3 One or more group substitutions of alkylacyl, aminoacyl, _C1-3 alkylaminoacyl, sulfonyl, sulfinyl, mercapto, aryl and heteroaryl _; and R5 is selected from H, _{C1- 6} alkyl, C _3-8 cycloalkyl and aryl, wherein said alkyl, cycloalkyl and aryl are optionally selected from C _1-6 alkyl, C _3-8 cycloalkyl, C _{3 -8} Heterocycloalkyl, C _1-6 alkoxy, C _1-6 alkylamino, halogen, hydroxyl, amino, nitro, cyano, C _1-6 alkyl acyl, aminoacyl, C _1-6 One or more groups in alkylaminoacyl, sulfonyl, sulfinyl, mercapto, aryl and heteroaryl are substituted, preferably, R ₅ is selected from H, C _1-3 alkyl, C _3-6 ring Alkyl and aryl, wherein said alkyl is optionally selected from C _1-3 alkyl, C _3-6 cycloalkyl, C _3-6 heterocycloalkyl, C _1-3 alkoxy, C _1-3 alkylamino, halogen, hydroxyl, amino, nitro, cyano, _C1-3 alkylacyl, aminoacyl, _C1-3 alkylaminoacyl, sulfonyl, sulfinyl, mercapto, aryl and One or more groups in the heteroaryl group are substituted.

In some preferred embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R _a1 is selected from C _4-8 heteroaryl , C _4-8 heterocycloalkyl and C _4-8 heterocycloalkenyl, wherein said heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxy, amino, C _{1- 6} alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkane oxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro, One or more groups in the cyano group are substituted;

R _a2 is selected from hydrogen, halogen, hydroxyl, amino, C _1-6 alkyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro and cyano; or

R _a1 , R _a2 together with the carbon atom to which they are attached constitute a C _4-8 heteroaryl, C _4-8 heterocycloalkyl or C _4-8 heterocycloalkenyl.

In other preferred embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R _a1 is selected from those containing at least one N atom Five- or six-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein said heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxy, amino, C _{1- 6} alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkane oxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro, One or more groups in the cyano group are substituted;

R _a2 is selected from hydrogen, halogen, hydroxyl, amino, C _1-6 alkyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro and cyano; or

R _a1 , R _a2 together with the carbon atom to which they are attached constitute a five- or six-membered heteroaryl, heterocycloalkyl or heterocycloalkenyl group containing at least one N atom. In other preferred embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R _a1 is selected from the group consisting of 1-3 N Atoms of five- or six-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein said heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxy, amino, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _{1- 6} alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro One or more groups in cyano group and cyano group are substituted;

R _a2 is selected from hydrogen, halogen, hydroxyl, amino, C _1-6 alkyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro and cyano; or

R _a1 , R _a2 together with the carbon atoms to which they are attached form a five- or six-membered heteroaryl, heterocycloalkyl or heterocycloalkenyl group containing 1-3 N atoms.

In other preferred embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R _a1 is selected from those containing only one S atom Five- or six-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein said heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxy, amino, C _{1- 6} alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkane oxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro, One or more groups in the cyano group are substituted;

R _a2 is selected from hydrogen, halogen, hydroxyl, amino, C _1-6 alkyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro and cyano; or

R _a1 , R _a2 together with the carbon atom to which they are attached form a five- or six-membered heteroaryl, heterocycloalkyl or heterocycloalkenyl group containing only one S atom.

In some specific embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R _a1 is selected from the group consisting of six atoms containing 1 N atom. membered heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein said heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxy, amino, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, C _{Of 1-6} alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro, cyano One or more groups are substituted; and

R _a2 is selected from hydrogen, halogen, hydroxyl, amino, C _1-6 alkyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro and cyano.

In other specific embodiments, the present invention provides compounds of general formula Ia, or stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or crystals thereof, wherein R _a1 , R _a2 and the carbon to which they are attached atoms together to form a five- or six-membered heteroaryl, heterocycloalkyl or heterocycloalkenyl containing 1 N atom, wherein said heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from Halogen, hydroxyl, amino, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy group, halogenated C _1-6 alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, One or more groups of sulfinyl, mercapto, nitro, and cyano are substituted.

In other specific embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R _a1 is selected from the group consisting of 2 N atoms Five-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein said heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxy, amino, C _1-6 alkyl , C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro, cyano substituted with one or more groups;

R _a2 is selected from hydrogen, halogen, hydroxyl, amino, C _1-6 alkyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro and cyano; or

R _a1 , R _a2 together with the carbon atoms to which they are attached form a five-membered heteroaryl, heterocycloalkyl or heterocycloalkenyl group containing 2 N atoms.

In other specific embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R _a1 is selected from containing 1 N atom and Five-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl with 1 sulfur atom, wherein said heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxyl, amino, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _{1- 6} alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro One or more groups in cyano group and cyano group are substituted;

R _a2 is selected from hydrogen, halogen, hydroxyl, amino, C _1-6 alkyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro and cyano; or

R _a1 , R _a2 together with the carbon atoms to which they are attached form a five-membered heteroaryl, heterocycloalkyl or heterocycloalkenyl group containing 1 N atom and 1 sulfur atom.

In other specific embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R _a1 is selected from containing 1 N atom and Five-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl with 1 oxygen atom, wherein said heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxyl, amino, C _1-6 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _{1- 6} alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro One or more groups in cyano group and cyano group are substituted;

R _a2 is selected from hydrogen, halogen, hydroxyl, amino, C _1-6 alkyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro and cyano; or

R _a1 , R _a2 together with the carbon atoms to which they are attached constitute a five-membered heteroaryl, heterocycloalkyl or heterocycloalkenyl group containing 1 N atom and 1 oxygen atom.

In other specific embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R _a1 is selected from containing only 1 S atom The five-membered heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein the heteroaryl, heterocycloalkyl and heterocycloalkenyl are optionally selected from halogen, hydroxyl, amino, C _1-6 alkane base, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy , C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro, cyano One or more groups in the substituted;

R _a2 is selected from hydrogen, halogen, hydroxyl, amino, C _1-6 alkyl, C _3-6 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 Alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylamino acyl, sulfonyl, sulfinyl, mercapto, nitro and cyano; or

R _a1 , R _a2 together with the carbon atoms to which they are attached form a five-membered heteroaryl, heterocycloalkyl or heterocycloalkenyl group containing only 1 S atom.

In some preferred embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R ₁ is H, methyl, ethyl, n-propyl or isopropyl, R ₂ is F, R ₃ is OH, R ₄ is selected from H, methyl, ethyl, n-propyl or isopropyl, R ₅ is selected from methyl, ethyl, n-propyl base, isopropyl, cyclopropyl, n-butyl, isobutyl, tert-butyl, cyclobutyl, n-pentyl, isopentyl, cyclopentyl or benzyl, R _a1 is selected from furan ring, pyrrole ring , thiophene ring, imidazole ring, pyrazole ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole ring, triazole ring, thiadiazole ring, oxadiazole ring, pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring, benzodiazepine, benzoxane, benzothiazepine, benzodiazepine, benzodioxane, benzodiazepine, benzoxazepine Ring, benzothiazepine heterocycle, wherein said R _a1 is optionally selected from halogen, hydroxyl, C _1-6 alkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl, amino acyl, C _1-6 alkyl amino acyl, nitro, One or more groups in the cyano group are substituted; R _a2 , R _a3 , R _a4 , R _a5 are independently selected from hydrogen, halogen, hydroxyl, amino, alkyl, C _3-8 cycloalkyl, halogenated C _{1 -6} alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylaminoacyl, sulfonyl, sulfinyl, mercapto, nitro, cyano.

In some preferred embodiments, the present invention provides a compound of general _formula Ia, or _a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R1 is H or methyl and R2 is F, R ₃ is OH, R ₄ is selected from H, methyl, ethyl, n-propyl or isopropyl, R ₅ is selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-propyl Butyl, isobutyl, tert-butyl, cyclobutyl, n-pentyl, isopentyl, cyclopentyl or benzyl, R _a1 is selected from furan ring, pyrrole ring, thiophene ring, imidazole ring, pyrazole ring, Thiazole ring, isothiazole ring, oxazole ring, isoxazole ring, triazole ring, thiadiazole ring, oxadiazole ring, pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring, benzofuran ring, benzopyrrole ring, benzothiophene ring, benzimidazole ring, benzopyrazole ring, benzothiazole ring, benzisothiazole ring, benzoxazole ring, benzisoxazole ring, benzotriazole ring, benzothiadiazole ring, benzoxadiazole ring, benzopyridine ring, benzopyrimidine ring, benzopyrazine ring, benzopyridazine ring, wherein said R _a1 is optionally selected from halogen , hydroxy, C _1-6 alkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1- Substituted by one or more groups in ₆ alkyl acylamino, C _1-6 alkyl acyl, aminoacyl, C _1-6 alkyl amino acyl, nitro, cyano; R _a2 , R _a3 , R _a4 , R _a5 is independently selected from hydrogen, halogen, hydroxyl, amino, alkyl, C _3-8 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy base, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylaminoacyl, sulfonyl, sulfinyl, mercapto, nitro, cyano group.

In some preferred embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R ₁ is H, methyl, ethyl, n-propyl or isopropyl, R ₂ is F, R ₃ is OH, R ₄ is selected from H, methyl, ethyl, n-propyl or isopropyl, R ₅ is selected from methyl, ethyl, n-propyl base, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl or benzyl, R _a1 , R _a2 together with the carbon atoms to which they are attached form benzazepine, benzo Oxygen heterocycle, benzothiazepine, benzodiazepine, benzodioxane, benzodiazepine, benzothiazepine, benzothiazepine, wherein the benzene Benzodiazepine, benzoxazepine, benzothiazepine, benzodiazepine, benzodiazepine, benzodiazepine, benzoxazepine, benzothiazepine Ring is optionally selected from halogen, hydroxy, _C1-6 alkyl, halogenated _C1-6 alkyl, _C1-6 alkoxy, halogenated _C1-6 alkoxy, amino, _C1-6 Substitution of one or more groups in alkylamino, C _{1-6 alkyl acylamino, C 1-6 alkyl} acyl, aminoacyl, C _1-6 alkyl amino acyl, nitro, and cyano; R _a3 , R _a4 , R _a5 are independently selected from hydrogen, halogen, hydroxyl, amino, alkyl, C _3-8 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _{1 -6} alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl, C _1-6 alkoxy acyl, aminoacyl, C _1-6 alkylaminoacyl, sulfonyl, sulfinyl, mercapto , nitro, cyano.

In some preferred embodiments, the present invention provides a compound of general formula Ia, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal thereof, wherein R ₁ is H, methyl, ethyl, n-propyl or isopropyl, R ₂ is F, R ₃ is OH, R ₄ is selected from H, methyl, ethyl, n-propyl or isopropyl, R ₅ is selected from methyl, ethyl, n-propyl base, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isoamyl or benzyl, R _a1 , R _a2 together with the carbon atoms to which they are attached form a benzofuran ring, benzopyrrole Ring, benzothiophene ring, benzimidazole ring, benzopyrazole ring, benzothiazole ring, benzisothiazole ring, benzoxazole ring, benzisoxazole ring, benzotriazole ring, benzene Thiadiazole ring, benzoxadiazole ring, benzopyridine ring, benzopyrimidine ring, benzopyrazine ring, benzopyridazine ring, wherein said benzofuran ring, benzopyrrole ring, benzene ring thiophene ring, benzimidazole ring, benzopyrazole ring, benzothiazole ring, benzisothiazole ring, benzoxazole ring, benzisoxazole ring, benzotriazole ring, benzothiadi azole ring, benzoxadiazole ring, benzopyridine ring, benzopyrimidine ring, benzopyrazine ring, benzopyridazine ring is optionally selected from halogen, hydroxyl, C _1-6 alkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, amino, C _1-6 alkylamino, C _1-6 alkyl acylamino, C _1-6 alkyl acyl , aminoacyl, C _1-6 alkylaminoacyl, nitro, cyano group is substituted with one or more groups; R _a3 , R _a4 , R _a5 are independently selected from hydrogen, halogen, hydroxyl, amino, alkyl , C _3-8 cycloalkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, C _1-6 alkylamino, C _1-6 alkyl acyl , C _1-6 alkoxyacyl, aminoacyl, C _1-6 alkylaminoacyl, sulfonyl, sulfinyl, mercapto, nitro, cyano.

In the above-mentioned preferred embodiments, the compounds of general formula Ia of the present invention are not the following compounds:

(2S)-2-(((4-(1,2,3-thiadiazol-4-yl)phenyl-1-yl)oxy)(((2R,3R,4R,5R)-5-( 2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino ) isopropyl propionate;

(2S)-2-(((2-(quinoxalin-5-yl)phenyl-1-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl;

(2S)-2-(((4-(thiazol-2-yl)phenyl-1-yl)oxy)(((2R,3R,4R,5R)-5-(2,4-dioxo- Isopropyl 3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate;

(2S)-2-(((4-(thiazol-5-yl)phenyl-1-yl)oxy)(((2R,3R,4R,5R)-5-(2,4-dioxo- Isopropyl 3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate;

(2S)-2-(((1H-Indol-5-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidine Isopropyl 1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate;

(2S)-2-(((4-Fluoro-2-methyl-1H-indol-5-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate ;

(2S)-2-(((3-methoxycarbonylmethyl-benzofuran-7-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo- Isopropyl 3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate;

(2S)-2-(((4-oxo-2-phenyl-4H-benzopyran-6-yl)oxy)(((2R,3R,4R,5R)-5-(2, 4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propane isopropyl acid;

(2S)-2-(((quinoxalin-5-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidine- Isopropyl 1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate;

(2S)-2-(((4-oxo-2-phenylchroman-7-yl)oxy)(((2R,3R,4R,5R)-5-(2,4 -Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionic acid isopropyl ester;

(2S)-2-((((4-(1H-1,2,4-triazol-1-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2, 4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propane isopropyl acid;

(2S)-2-(((4-(pyrimidin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4- Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl;

(2S)-2-(((S)-((4-Fluoro-1,2-dimethyl-1H-indol-5-yl)oxy)(((2R,3R,4R,5R)- 5-(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphorus Acyl)amino)isopropyl propionate;

(2S)-2-((((6-Fluoro-3-methylbenzo[d]isoxazol-5-yl)oxy)(((2R,3R,4R,5R)-5-(2 ,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino) isopropyl propionate;

(2S)-2-((((6-Fluorobenzo[c][1,2,5]thiadiazol-5-yl)oxy)(((2R,3R,4R,5R)-5- (2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl) amino) isopropyl propionate; and

(2S)-2-(((4-(thiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4- Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionic acid isopropyl ester.

The present invention provides the following specific compounds:

On the other hand, the present invention provides the preparation method of the compound of general formula I of the present invention, the method comprises the following steps:

a) The compound of formula (1) is reacted with phosphorus oxychloride under basic conditions to prepare the compound of formula (2);

b) the compound of formula (2) is reacted with the compound of formula (3) to obtain the compound of formula (4);

c) The compound of formula (4) is reacted with pentafluorophenol to obtain the compound of formula (5);

d) Compounds of formula (5) are reacted with compounds of formula (6) to produce compounds of general formula (I).

wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , Cy ₁ , Cy ₂ are as defined in general formula I above.

In a third aspect, the present invention provides a pharmaceutical composition comprising the compound represented by the general formula I or Ia, a stereoisomer, a pharmaceutically acceptable salt, a hydrate, a solvate or a crystal of the present invention.

In some embodiments, the present invention provides compounds, stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or crystals of the present invention and compounds, stereoisomers, pharmaceutically acceptable salts comprising the present invention , hydrate, solvate or crystalline pharmaceutical composition for the treatment and/or prevention of liver disease caused by hepatitis C virus.

In some embodiments, the present invention provides a pharmaceutical composition comprising a compound, stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal of Formula I or Ia, and further comprising a composition selected from the group consisting of: One or more additional anti-HCV therapeutics of: HCV NS3 protease inhibitor, HCV NS5B RNA-dependent RNA polymerase inhibitor, nucleoside analog, interferon alpha, pegylated interferon, ribavir Lin, levovirine, viramidine, TLR7 agonists, TLR9 agonists, cyclophilin inhibitors, alpha glucosidase inhibitors, NS5A inhibitors, and NS3 helicase inhibitors.

Compounds, stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or crystals of the present invention shown in general formula I and Ia can be mixed with pharmaceutically acceptable carriers, diluents or excipients to prepare. Pharmaceutical formulations suitable for oral or parenteral administration. Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, and oral routes. The formulations may be administered by any route, such as by infusion or bolus injection, by route of absorption through the epithelium or mucocutaneous (eg, oral mucosa or rectum, etc.). Administration can be systemic or local. Examples of formulations for oral administration include solid or liquid dosage forms, specifically, tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions and the like. The formulations can be prepared by methods known in the art and include carriers, diluents or excipients conventionally used in the art of pharmaceutical formulations.

In the fourth aspect, the present invention provides the compounds, stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or crystals of the present invention shown in general formulas I and Ia or the pharmaceutical compositions of the present invention to treat Flaviviridae viruses A method for an infected subject comprising administering to said subject a compound, stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystal of or comprising formula I and Ia The compound, stereoisomer, pharmaceutically acceptable salt, hydrate, solvate or crystalline pharmaceutical composition of the compound is administered in an amount effective to reduce the viral load of the virus in the subject. In one embodiment, the present invention provides a method for the treatment and/or prevention of infection by an RNA virus, such as a Flaviviridae virus, comprising administering to an individual in need of such treatment a compound of the present invention, a stereoisomer thereof, a pharmaceutically acceptable Salts, hydrates, solvates or crystals or pharmaceutical compositions thereof. In another embodiment, the present invention provides a method of inhibiting infection by an RNA virus, such as a Flaviviridae virus, comprising hydrating the virus with a therapeutically effective amount of a compound of the present invention, a stereoisomer, a pharmaceutically acceptable salt, compound, solvate or crystal or a pharmaceutical composition thereof.

"Flaviviridae" refers to any virus of the Flaviviridae family, including those viruses that infect humans and non-human animals, such as flaviviruses, plague virus, and hepatitis C virus. The compounds and compositions of the present invention are particularly useful in the therapeutic and/or prophylactic treatment of HCV.

On the other hand, the present invention provides compounds, stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or crystals of the present invention shown in general formula I and Ia for preventing or treating viral infections, especially yellow The application of viral infectious diseases, and the application in the preparation of medicines for preventing and/or treating viral infectious diseases, especially the application in preparing medicines for preventing and/or treating HCV viral infections, such as HCV viral hepatitis diseases. Examples of such diseases are acute hepatitis C, chronic hepatitis C and co-infection of hepatitis C and hepatitis B or D.

In some embodiments, the present invention provides methods for treating and/or preventing viral infections, the methods comprising administering to an individual in need thereof a therapeutically and/or prophylactically effective amount of a compound of the present invention or a stereoisomer thereof body, pharmaceutically acceptable salt, hydrate, solvate or crystal or the pharmaceutical composition of the present invention. The compounds of the present invention or their stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or crystals or the pharmaceutical compositions of the present invention can be administered to mammals in need thereof to inhibit the virus and prevent the progression of the disease course.

In other embodiments, the method or use of treating and/or preventing a viral infection further comprises administering to the individual a compound of formula I of the present invention or a stereoisomer, pharmaceutically acceptable salt, hydrate thereof , solvates or crystals or pharmaceutical compositions containing them, and in the administration of the compounds of formula I of the present invention or their stereoisomers, pharmaceutically acceptable salts, hydrates, solvates or crystals or drugs containing them At least one other compound having anti-HCV activity is administered before, after or concurrently with the composition.

In some embodiments, at least one of the other compounds is interferon or ribavirin. In some specific embodiments, the interferon is selected from the group consisting of interferon alpha 2B, PEGylated interferon alpha, syngeneic interferon, interferon alpha 2A, and lymphoblastoid interferon tau. In other embodiments, at least one of the other compounds is selected from the group consisting of interleukin 2, interleukin 6, interleukin 12, interfering RNA, antisense RNA, imiquimod, ribavirin, Inosine 5'-monophosphate dehydrogenase inhibitors, amantadine and rimantadine. In other embodiments, at least one of the other compounds is effective to inhibit the function of a target selected from the group consisting of HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV NS5B protein, HCV entry, HCV assembly, HCV release, HCV NS3/4A protein and IMPDH.

Definition of Terms

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

The term "stereoisomer" refers to isomers that result from different arrangements of atoms in a molecule in space. Including cis-trans isomers, enantiomers and conformational isomers. All stereoisomers are within the scope of the present invention. Individual stereoisomers of the compounds of the present invention may be substantially free of other isomers or may be mixed, for example, as racemates, or with all other stereoisomers.

The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt formed by the compound of the present invention and an acid, for example, the acid can be selected from, but not limited to: phosphoric acid, sulfuric acid, hydrochloric acid, hydrobromic acid, Citric acid, maleic acid, malonic acid, mandelic acid, succinic acid, fumaric acid, acetic acid, lactic acid, nitric acid, sulfonic acid, p-toluenesulfonic acid, malic acid, methanesulfonic acid, or the like.

The term "solvate" refers to a form of a compound of the present invention that forms a complex in a solid or liquid state by coordination with solvent molecules. Hydrates are a special form of solvates in which coordination occurs with water. In the context of the present invention, solvates are preferably hydrates.

The term "crystalline" refers to various solid forms formed by the compounds of the present invention, including crystalline forms and amorphous forms.

The term "alkyl" refers to a straight or branched chain saturated hydrocarbon group, preferably a hydrocarbon group having less than 6 carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, 2,2-methylbutyl and 2,3-dimethylbutyl. The term "C _1-6 alkyl" refers to a straight or branched chain saturated hydrocarbon group containing 1 to 6 carbon atoms. The term "C _1-4 alkyl" refers to a straight or branched chain saturated hydrocarbon group containing 1 to 4 carbon atoms.

The term "haloalkyl" refers to an alkyl group substituted with at least one halogen atom.

The term "cycloalkyl" refers to a cyclic saturated hydrocarbon group, preferably a hydrocarbon group of 8 or less carbon atoms. The term "C _3-8 cycloalkyl" refers to a cyclic saturated hydrocarbon group containing 3-8 carbon atoms. The term "C _3-6 cycloalkyl" refers to a straight or branched chain saturated hydrocarbon group containing 3 to 6 carbon atoms.

The term "alkoxy" refers to -O-alkyl.

The term "halogen" refers to fluorine, chlorine, bromine, iodine.

The term "alkylamino" refers to -NH-alkyl or -N-(alkyl)(alkyl).

The term "alkylacyl" refers to -C(O)-alkyl.

The term "aminoacyl" refers to -C(O) _-NH2 and the term "alkylaminoacyl" refers to -C(O)-NH-alkyl or -C(O)-N-(alkyl)(alkyl ).

The term "sulfonyl" refers to -S(O) ₂ -alkyl and the term "sulfinyl" refers to -S(O)-alkyl.

The term "aryl" refers to an aromatic hydrocarbon group containing one or more benzene rings. Suitable aryl groups include phenyl, naphthyl.

The term "heteroaryl" refers to an aromatic group in which at least one carbon atom in the aryl group is replaced by a heteroatom. Said heteroatoms are O, S, N. For example, heteroaryl groups described herein include, but are not limited to, pyridyl, pyrazolyl, oxazolyl, thiazolyl, thienyl, furyl, pyrrolyl, isoxazolyl, isothiazolyl, imidazolyl, triazolyl , tetrazolyl, oxadiazolyl, thiadiazolyl, pyrimidinyl, pyridazinyl, pyrazinyl and triazinyl.

The term "heterocycloalkyl" refers to a saturated cyclic group containing at least one heteroatom, wherein the heteroatom is N, O or S.

The term "heterocycloalkenyl" refers to an unsaturated cyclic group containing at least one heteroatom, wherein the heteroatom is N, O or S.

Detailed ways

The present invention will be described in further detail below in conjunction with the examples, but the present invention is not limited to these examples. The reagents and raw materials used in the present invention are all commercially available.

Example 1(2S)-2-(((4-(pyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3 ,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl ester

Step 1 Preparation of 4-(pyridin-2-yl)phenol

Weigh 1.38g of 4-hydroxyphenylboronic acid (10mmol), 1.58g of 2-bromopyridine (10mmol), 0.4g of [1,1'-bis(diphenylphosphorus)ferrocene]palladium dichloride (Pd(dppf) )Cl ₂ , 0.5mmol) and 9g cesium carbonate (30mmol) in a 250mL eggplant-shaped flask, add 50mL 1,4-dioxane and 5mL water, and react at 90°C for 1.5h under argon protection. After the reaction, Add 100 mL of ethyl acetate and 50 mL of saturated aqueous sodium chloride solution for extraction, collect the organic phase, wash with saturated aqueous sodium chloride solution (3×50 mL), dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure, and purify by column chromatography to obtain the title compound .

LC-MS m/z: [M+H] ⁺ =172.

Step 2 Preparation of isopropyl 2-(((pentafluorophenoxy)(4-(pyridin-2-yl)phenoxy)phosphoryl)amino)propionate

Measure 0.32 mL of phosphorus oxychloride into a reaction flask, add 5 mL of anhydrous tetrahydrofuran, and dropwise add 10 mL of a solution containing 342 mg of 4-(pyridin-2-yl)phenol obtained in step 1 and 0.42 mL of triethylamine at -60°C. Anhydrous tetrahydrofuran solution, after dripping, warmed up to 25°C for reaction for 2h, then added 334mg of alanine isopropyl ester hydrochloride at -60°C, stirred for 10min, and added dropwise 5mL of anhydrous tetrahydrofuran dissolved with 1.05mL of triethylamine , the dropwise addition was completed, the temperature was slowly raised to -5°C, 5 mL of anhydrous tetrahydrofuran liquid in which 0.385 g of pentafluorophenol and 0.42 mL of triethylamine were dissolved was added, the dropwise addition was completed, and the reaction was carried out at room temperature for 30 minutes. The title compound was obtained.

Step 3(2S)-2-(((4-(pyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3, Preparation of isopropyl 4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

Weigh 260mg (2'R)-2'-deoxy-2'-fluoro-2'-methyluridine (1mmol) in a reaction bottle, seal it, inject 10mL anhydrous tetrahydrofuran (THF) under argon protection, 0 ℃, inject 10 mL of t-BuMgCl solution (1M in THF), and after 3 hours of reaction at room temperature, inject 10 mL of 2-(((pentafluorophenoxy)(4-(pyridin-2-yl)benzene dissolved in 600 mg of step 2) The tetrahydrofuran solution of isopropyl oxy)phosphoryl)amino)propionate was reacted at 25°C for 16h. After the reaction was completed, 15mL of 2N HCl solution was added to quench the reaction, extracted with ethyl acetate, and the organic phase was washed with saturated sodium bicarbonate solution, Dry, concentrate under reduced pressure, and purify by column chromatography to obtain the title compound.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.51 (s, 1H), 8.64-8.65 (m, 1H), 8.08-8.11 (m, 2H), 7.93-7.95 (m, 2H), 7.56-7.59 (m,1H),7.32-7.35(m,3H),6.12-6.16(m,2H),5.84-5.87(m,1H),5.58(m,1H),4.87-4.89(m,1H),4.36 -4.38(m,1H),4.26-4.28(m,1H),4.01(m,1H),3.82-3.84(m,2H),1.26-1.29(m,6H),1.13-1.16(m,6H) .

LC-MS m/z: [M+H] ⁺ =607.

Example 2(2S)-2-(((4-(3-fluoropyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

Step 1 Preparation of 4-(3-fluoropyridin-2-yl)phenol

Using 4-hydroxyphenylboronic acid and 2-bromo-3-fluoropyridine as starting materials, the title compound was obtained by the same method as in Example 1, step 1.

LC-MS m/z: [M+H] ⁺ =190.

Step 2(2S)-2-(((4-(3-fluoropyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)isopropyl propionate preparation

4-(3-Fluoropyridin-2-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxygenate obtained in step 1 -2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300 MHz, DMSO-d ₆ ) δppm: 11.51 (s, 1H), 8.53-8.54 (m, 1H), 7.92-7.95 (m, 2H), 7.83-7.86 (m, 1H), 7.54-7.57 (m,1H),7.45-7.47(m,1H),7.33-7.35(m,2H),6.19-6.23(m,1H),6.07-6.15(m,1H),5.91-5.93(m,1H) ,5.57-5.60(m,1H),4.87-4.90(m,1H),4.41-4.46(m,1H),4.30-4.30(m,1H),4.05(m,1H),3.79-3.82(m, 2H), 1.20-1.27 (m, 6H), 1.13-1.17 (m, 6H).

LC-MS m/z: [M+H] ⁺ =625.

Example 3(2S)-2-(((4-(2-fluoropyridin-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

Step 1 Preparation of 4-(2-fluoropyridin-4-yl)phenol

Using 4-hydroxyphenylboronic acid and 2-fluoro-4-bromopyridine as starting materials, the title compound was obtained by the same method as Example 1, step 1.

LC-MS m/z: [M+H] ⁺ =190.

Step 2(2S)-2-(((4-(2-fluoropyridin-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)isopropyl propionate preparation

4-(2-Fluoropyridin-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxygenate obtained in step 1 -2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.51 (s, 1H), 8.28-8.30 (m, 1H), 7.89-7.92 (m, 2H), 7.68-7.70 (m, 1H), 7.55-7.58 (m,1H),7.52(m,1H),7.36-7.39(m,2H),6.11-6.19(m,1H),5.98-6.05(m,1H),5.84-5.86(m,1H),5.54 -5.56(m, 1H), 4.81-4.90(m, 1H), 4.37-4.39(m, 1H), 4.26-4.28(m, 1H), 3.89-4.01(m, 1H), 3.83-3.85(m, 2H), 1.22-1.29 (m, 6H), 1.14-1.16 (m, 6H).

LC-MS m/z: [M+H] ⁺ =625.

Example 4(2S)-2-(((4-(pyridin-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3 ,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl ester

Step 1 Preparation of 4-(pyridin-4-yl)phenol

Using 4-hydroxyphenylboronic acid and 4-bromopyridine as starting materials, the title compound was prepared in the same manner as in Example 1, step 1. LC-MS m/z: [M+H] ⁺ =172.

Step 2(2S)-2-(((4-(pyridin-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3, Preparation of isopropyl 4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

4-(Pyridin-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy-2' obtained in step 1 -Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d ₆ ) δppm: 11.52 (s, 1H), 8.61-8.62 (m, 2H), 7.82-7.84 (m, 2H), 7.67-7.68 (m, 2H), 7.36-7.37 (m,1H),7.32-7.34(m,2H),6.19-6.21(m,1H),6.13-6.16(m,1H),5.90-5.91(m,1H),5.56-5.58(m,1H) ,4.83-4.86(m,1H),4.30-4.31(m,1H),4.26-4.29(m,1H),4.01-4.05(m,1H),3.82-3.84(m,2H),1.26-1.28( m, 6H), 1.13-1.15 (m, 6H).

LC-MS m/z: [M+H] ⁺ =607.

Example 5(2S)-2-(((4-(1-methyl-1H-pyrazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2 ,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino) isopropyl propionate

Step 1 4-(1-Methyl-1H-pyrazol-4-yl)phenol

Using 4-hydroxyphenylboronic acid and 1-methyl-4-bromo-1H-pyrazole as starting materials, the title compound was obtained by the same method as Example 1, step 1.

LC-MS m/z: [M+H] ⁺ =175.

Step 2(2S)-2-(((4-(1-methyl-1H-pyrazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2, 4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propane Preparation of isopropyl acid

4-(1-Methyl-1H-pyrazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl hydrochloride, pentafluorophenol and (2'R) obtained in step 1 -2'-Deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.49(s,1H), 8.06(s,1H), 7.79(m,1H), 7.52-7.58(m,3H), 7.15-7.17(m,2H) ),6.00-6.06(m,2H),5.86-5.88(m,1H),5.55-5.60(m,1H),4.83-4.90(m,1H),4.38-4.42(m,1H),4.27-4.29 (m,1H),4.03-4.06(m,1H),3.81-3.83(m,3H),3.78-3.81(m,2H),1.20-1.28(m,6H),1.14-1.17(m,6H) .

LC-MS m/z: [M+H] ⁺ =610.

Example 6(2S)-2-(((4-(1-methyl-1H-pyrazol-3-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2 ,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino) isopropyl propionate

Step 1 4-(1-Methyl-1H-pyrazol-3-yl)phenol

Using 4-hydroxyphenylboronic acid and 3-bromo-1-methyl-1H-pyrazole as starting materials, the title compound was obtained by the same method as Example 1, step 1.

LC-MS m/z: [M+H] ⁺ =175.

Step 2(2S)-2-(((4-(1-Methyl-1H-pyrazol-3-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2, 4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propane Preparation of isopropyl acid

4-(1-Methyl-1H-pyrazol-3-yl)phenol, phosphorus oxychloride, L-alanine isopropyl hydrochloride, pentafluorophenol and (2'R) obtained in step 1 -2'-Deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.49(s,1H), 8.06(s,1H), 7.79(m,1H), 7.52-7.58(m,3H), 7.15-7.17(m,2H) ),6.00-6.06(m,2H),5.86-5.88(m,1H),5.55-5.60(m,1H),4.83-4.90(m,1H),4.38-4.42(m,1H),4.27-4.29 (m,1H),4.03-4.06(m,1H),3.81-3.83(m,3H),3.78-3.81(m,2H),1.20-1.28(m,6H),1.14-1.17(m,6H) .

LC-MS m/z: [M+H] ⁺ =610.

Example 7(2S)-2-(((4-(1-methyl-3-methylaminocarbonyl-1H-pyrazol-4-yl)phenyloxy)(((2R,3R,4R,5R )-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy )phosphoryl)amino)isopropyl propionate

Step 1 Methyl 4-bromo-1-methyl-1H-pyrazole-3-carboxylate

Weigh 4.10g of 4-bromo-1-methyl-1H-pyrazole-3-carboxylic acid (20mmol) into a 250mL eggplant flask, add 100ml methanol, and slowly add 5ml thionyl chloride at 0-4°C , after the reaction at room temperature, 500 mL of ethyl acetate and 250 mL of saturated aqueous sodium chloride were added for extraction, the organic phase was collected, washed with water 3 times, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound.

LC-MS m/z: [M+H] ⁺ =219.

Step 2 4-(1-Methyl-3-methoxycarbonyl-1H-pyrazol-4-yl)phenol

Using methyl 4-bromo-1-methyl-1H-pyrazole-3-carboxylate obtained in step 1 and 4-hydroxybenzeneboronic acid as starting materials, the title compound was prepared in the same manner as in step 1 of Example 1.

LC-MS m/z: [M+H] ⁺ =233.

Step 3 4-(1-Methyl-3-methylaminocarbonyl-1H-pyrazol-4-yl)phenol

Weigh 2.33 g of 4-(1-methyl-3-methoxycarbonyl-1H-pyrazol-4-yl)phenol (10 mmol) obtained in step 2 into a 250 mL eggplant-shaped flask, add 100 mL of methyl ammonia solution and 10ml of methanol was reacted at room temperature. After the reaction was completed, 500ml of ethyl acetate was added, and 250ml of saturated aqueous sodium chloride solution was added for extraction. The organic phase was collected, washed three times with water, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title. compound.

LC-MS m/z: [M+H] ⁺ =232.

Step 4(2S)-2-(((4-(1-Methyl-3-methylaminocarbonyl-1H-pyrazol-4-yl)phenyloxy)(((2R,3R,4R,5R) -5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy) Phosphoryl)amino)isopropyl propionate

Using the product obtained in step 3, 4-(1-methyl-3-methylaminocarbonyl-1H-pyrazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol Using (2'R)-2'-deoxy-2'-fluoro-2'-methyluridine as a raw material, the target compound was prepared in the same manner as in steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.45(s, 1H), 8.03-8.04(m, 1H), 7.95(m, 1H), 7.53-7.56(m, 3H), 7.11-7.14(m ,2H),6.06-6.10(m,2H),5.88-5.98(m,1H),5.56-5.59(m,1H),4.85-4.87(m,1H),4.38-4.39(m,1H),4.25 -4.29(m, 1H), 4.02-4.04(m, 1H), 3.81-3.87(m, 3H), 3.72-3.78(m, 2H), 2.86-2.70(m, 3H), 1.21-1.26(m, 6H), 1.13-1.16 (m, 6H).

LC-MS m/z: [M+H] ⁺ =667.

Example 8(2S)-2-(((4-(1-methyl-3-cyano-1H-pyrazol-4-yl)phenyloxy)(((2R,3R,4R,5R) -5-(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy) Phosphoryl)amino)isopropyl propionate

Step 1 Preparation of 1-methyl-3-aminoacyl-4-bromo-1H-pyrazole

Take 4.10g 4-bromo-1-methyl-1H-pyrazole-3-carboxylic acid (20mmol), 2.14g ammonium chloride (40mmol), 15.2g HATU (40mmol) and 6.98ml DIPEA (40mmol) in 250mL of eggplant 100ml of DMF was added to the flask, and the reaction was carried out at room temperature for 12h. After the reaction was completed, 500ml of ethyl acetate and 250ml of saturated aqueous sodium chloride were added for extraction. The organic phase was collected, washed three times with water, dried over anhydrous sodium sulfate, filtered, concentrated, and the column layer was The title compound was obtained by analytical purification.

LC-MS m/z: [M+H] ⁺ =204.

Step 2 Preparation of 1-methyl-3-cyano-4-bromo-1H-pyrazole

Weigh 2.03 g of 1-methyl-3-aminoacyl-4-bromo-1H-pyrazole (10 mmol) obtained in step 1 into a 250 mL eggplant flask, add 100 mL of pyridine, and slowly add 30 mL of pyridine at 0-4°C Phosphorus oxychloride was reacted at room temperature for 12 hours. After the reaction, saturated aqueous sodium carbonate solution was added to neutralize, and then 500 mL of ethyl acetate and 250 mL of saturated aqueous sodium chloride solution were added for extraction. The organic phase was collected, washed with water 3 times, and dried over anhydrous sodium sulfate. , filtered, concentrated, and purified by column chromatography to obtain the title compound.

LC-MS m/z: [M+H] ⁺ =186.

Step 3 4-(1-Methyl-3-cyano-1H-pyrazol-4-yl)phenol

Using 4-hydroxyphenylboronic acid and 1-methyl-3-cyano-4-bromo-1H-pyrazole obtained in step 2 as starting materials, the title compound was prepared in the same manner as in step 1 of Example 1.

LC-MS m/z: [M+H] ⁺ =200.

Step 4(2S)-2-(((4-(1-methyl-3-cyano-1H-pyrazol-4-yl)phenyloxy)(((2R,3R,4R,5R)- 5-(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphorus Acyl)amino)isopropyl propionate

Using the resultant of step 3, 4-(1-methyl-3-cyano-1H-pyrazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300 MHz, DMSO-d ₆ ) δppm: 11.49 (s, 1H), 8.03-8.09 (m, 1H), 7.61-7.62 (m, 2H), 7.55-7.57 (m, 1H), 7.33-7.34 (m,2H),6.09-6.11(m,1H),6.04-6.07(m,1H),5.98-6.00(m,1H),5.54-5.56(m,1H),4.83-4.88(m,1H) ,4.38-4.40(m,1H),4.24-4.27(m,1H),4.01-4.03(m,1H),3.82-3.84(m,3H),3.81(m,2H),1.23-1.27(m, 6H), 1.15-1.15 (m, 6H).

LC-MS m/z: [M+H] ⁺ =635.

Example 9(2S)-2-(((4-(pyridin-3-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3 ,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl ester

Step 1 Preparation of 4-(pyridin-3-yl)phenol

In a 100mL single-neck flask, add 1g 3-bromopyridine (6.4mmol), 0.96g p-hydroxyphenylboronic acid (7.0mmol), 2.6g potassium carbonate (21mmol), 0.45g PdCl ₂ (PPh ₃ ) ₂ (0.65mmol), 15mL Water and 30 mL of 1,4-dioxane were reacted at 80°C for 2 h under nitrogen protection. After the reaction was completed, cooled to room temperature, filtered, concentrated, extracted with ethyl acetate and water, the organic phase was collected, dried over anhydrous sodium sulfate, Filtration, concentration, and purification by column chromatography gave the title compound.

¹ HNMR ((300MHz, DMSO-d6) δppm: 9.67(s, 1H), 8.82(s, 1H), 8.49(d, 1H), 7.99(d, 1H), 7.65-7.54(m, 2H), 7.46 -7.42(m, 1H), 6.88(d, 2H).

LC-MS m/z: [M+H] ⁺ =172.

Step 2(2S)-2-(((4-(pyridin-3-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3, Isopropyl 4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

4-(Pyridin-3-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy-2' obtained in step 1 -Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.50(s, 1H), 8.87(s, 1H), 8.56(d, 1H), 8.05(d, 1H), 7.74(d, 2H), 7.59- 7.46(m, 2H), 7.35(d, 2H), 6.20-6.00(m, 2H), 5.90(d, 1H), 5.58(d, 1H), 4.90-4.82(m, 1H), 4.43(m, 1H), 4.31 (m, 1H), 4.05 (m, 1H), 3.84-3.78 (m, 2H), 1.28-1.20 (m, 6H), 1.17-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =607.

Example 10(2S)-2-(((4-(5-fluoropyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

Step 1 Preparation of 4-(5-fluoropyridin-2-yl)phenol

Using 2-bromo-5-fluoropyridine and 4-hydroxybenzeneboronic acid as starting materials, the title compound was prepared in the same manner as in Example 9, step 1.

¹ HNMR ((300MHz, CDCl3-d ₆ ) δppm: 8.50(s, 1H), 7.82(d, 2H), 7.64(m, 1H), 7.46(m, 1H), 6.90(d, 2H), 7.53( s, 1H).

Step 2(2S)-2-(((4-(5-fluoropyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl ester

4-(5-Fluoropyridin-2-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxygenate obtained in step 1 -2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.50(s, 1H), 8.64(d, 1H), 8.04(d, 3H), 7.80(m, 1H), 7.56(d, 1H), 7.33( d,2H),6.15-6.07(m,2H),5.84(d,1H),5.56(d,1H),4.90-4.83(m,1H),4.43(m,1H),4.27(m,1H) , 4.01 (m, 1H), 3.88-3.78 (m, 2H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =625.

Example 11(2S)-2-(((4-(2-chloropyridin-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

Step 1 Preparation of 4-(2-chloropyridin-4-yl)phenol

Using 2-chloro-4-bromopyridine and p-hydroxybenzeneboronic acid as starting materials, the title compound was prepared by the same method as in Example 9, step 1.

¹ HNMR ((300MHz, CDCl3-d ₆ ) δppm: 8.50(s, 1H), 7.82(d, 2H), 7.64(m, 1H), 7.46(m, 1H), 6.90(d, 2H), 7.53( s, 1H).

LC-MS m/z: [M+H] ⁺ =206.

Step 2(2S)-2-(((4-(2-chloropyridin-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl ester

4-(2-Chloropyridin-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxygenate obtained in step 1 -2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.51(s, 1H), 8.46(d, 1H), 7.92(m, 3H), 7.42(d, 1H), 7.62-7.56(m, 1H), 7.37(d, 2H), 6.20-5.99(m, 2H), 5.86(d, 1H), 5.54(d, 1H), 4.88-4.81(m, 1H), 4.39(m, 1H), 4.27(m, 1H), 4.04 (m, 1H), 3.89-3.81 (m, 2H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =641.

Example 12(2S)-2-(((4-(5-methylthiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Isopropyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate ester

Step 1 Preparation of 4-bromo-5-methylthiazole

1g 5-methylthiazole (10mmol), 1.8g N-bromosuccinimide (10mmol) and 30mL dichloromethane were added in a 100mL single-neck flask, and the reaction was carried out at room temperature for 12h. After the reaction was completed, it was concentrated and purified by column chromatography. The title compound was obtained.

Step 2 Preparation of 4-(5-methylthiazol-4-yl)phenol

Using 4-bromo-5-methylthiazole obtained in step 1 and p-hydroxyphenylboronic acid as starting materials, the title compound was prepared in the same manner as in step 1 of Example 9.

¹ H NMR ((300 MHz, CDCl ₃ -d6) δ ppm: 8.65 (s, 1H), 7.52 (d, 2H), 6.88 (d, 2H), 2.57 (s, 3H).

Step 3(2S)-2-(((4-(5-Methylthiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

4-(5-methylthiazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'- Deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.49(s, 1H), 7.89(d, 2H), 7.58(m, 2H), 7.33(d, 2H), 6.18-5.98(m, 2H), 5.86(d,1H),5.57(d,1H),4.88-4.81(m,1H),4.39(m,1H),4.27(m,1H),4.04(m,1H),3.89-3.81(m, 2H), 2.50 (s, 3H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =627.

Example 13(2S)-2-(((1-methyl-1H-indazol-5-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl ester

With 5-hydroxy-1-methyl-1H-indazole, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy-2'- Fluoro-2'-methyluridine was used as the raw material, and the target compound was prepared in the same manner as in steps 2 and 3 of Example 1.

1H NMR (300MHz, DMSO-d ₆ ) δppm: 11.49(s, 1H), 7.99(s, 1H), 7.63(d, 1H), 7.54(d, 2H), 7.26(d, 1H), 6.18-5.98 (m, 2H), 5.86(d, 1H), 5.57(d, 1H), 4.88-4.81(m, 1H), 4.39(m, 1H), 4.27(m, 1H), 4.04(m, 1H), 3.99(s, 3H), 3.89-3.81(m, 2H), 1.29-1.22(m, 6H), 1.16-1.14(m, 6H).

LC-MS m/z: [M+H] ⁺ =584.

Example 14(2S)-2-(((1-methyl-1H-indol-5-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl ester

Step 1 Preparation of 5-methoxy-1-methyl-1H-indole

Weigh 5g of 5-methoxy-1H-indole (0.034mol) into a 500mL single-neck flask, add 200mL of DMF to dissolve, add 2.7g of sodium hydride (0.068mol) at 0-4°C, and stir for 0.5h after the addition is complete. , 4.4g of dimethyl sulfate (0.035mol) was added dropwise, the addition was completed, and the reaction was carried out at room temperature for 0.5h. After the reaction was completed, water was added under an ice bath to quench, filtered, and dried to obtain the title compound.

Step 2 Preparation of 5-hydroxy-1-methyl-1H-indole

Weigh 0.7 g of 5-methoxy-1-methyl-1H-indole (4.34 mmol) obtained in step 1 into a 100 mL single-neck flask, add 20 mL of dichloromethane to dissolve, and dropwise add 0.52 g of tribromide at -30°C Boronide (21 mmol) was added, and stirred at room temperature for 12 h. After the reaction was completed, dichloromethane was added to dilute in an ice bath, quenched by adding water, extracted, dried, concentrated, and purified by column chromatography to obtain the title compound.

Step 3(2S)-2-(((1-Methyl-1H-indol-5-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo- Isopropyl 3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

5-Hydroxy-1-methyl-1H-indole, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxygenated from step 2 -2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.49(s, 1H), 8.37(d, 1H), 7.55-7.34(m, 3H), 7.02(d, 1H), 6.36(d, 1H), 6.04-5.85(m, 3H), 5.48(d, 1H), 4.88-4.81(m, 1H), 4.35(m, 1H), 4.22(m, 1H), 4.043.78(m, 1H), 3.89- 3.31 (m, 5H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =583.

Example 15(2S)-2-(((2-Fluoro-4-(1-methyl-1H-pyrazol-3-yl)phenyloxy)(((2R,3R,4R,5R)- 5-(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphorus Acyl)amino)isopropyl propionate

Step 1 Preparation of 4-(1-methyl-1H-pyrazol-3-yl)phenol

Using 1-methyl-3-bromo-1H-pyrazole and 4-hydroxybenzeneboronic acid as starting materials, the title compound was obtained by the same method as in Example 1, step 1.

LC-MS m/z: [M+H] ⁺ =175.

Step 2 Preparation of 2-fluoro-4-(1-methyl-1H-pyrazol-3-yl)phenol

Weigh 1.71g of the obtained 4-(1-methyl-1H-pyrazol-3-yl)phenol in step 1, 3.54g of 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2 .]Octane difluoroborate (selectfluor) was placed in a 100 mL eggplant-shaped flask, 50 mL of trifluoroacetic acid was added, and the reaction was carried out at 60°C for 12 h. After the reaction, the pH value of the reaction solution was adjusted to about 7, and 100 mL of ethyl acetate and ethyl acetate were added. Extracted with 50 mL of saturated aqueous sodium chloride solution, washed three times with water, collected the organic phase, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound.

LC-MS m/z: [M+H] ⁺ =193.

Step 3(2S)-2-(((2-Fluoro-4-(1-methyl-1H-pyrazol-3-yl)phenyloxy)(((2R,3R,4R,5R)-5 -(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl ) preparation of amino) isopropyl propionate

Using the resultant of step 2, 2-fluoro-4-(1-methyl-1H-pyrazol-3-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and ( 2'R)-2'-deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.50 (s, 1H), 7.70-7.73 (m, 2H), 7.60-7.66 (m, 2H), 7.44-7.47 (m, 1H), 6.67 (s) ,1H),6.21-6.24(m,1H),6.19-6.20(m,1H),5.86-5.89(m,1H),5.56-5.59(m,1H),4.81-4.90(m,1H),4.38 -4.42(m, 1H), 4.27-4.28(m, 1H), 4.01-4.06(m, 1H), 3.81-3.84(m, 3H), 3.78-3.80(m, 2H), 1.27-1.29(m, 6H), 1.14-1.16 (m, 6H).

LC-MS m/z: [M+H] ⁺ =628.

Example 16(2S)-2-(((4-(5-cyanothiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Isopropyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate ester

Step 1 Preparation of 4-bromo-5-methylthiazole

Weigh 15g of 5-methylthiazole into a 500mL single-necked bottle, add 250mL of dichloromethane to dissolve, add 27g of NBS in batches, complete the addition, and react at room temperature for 12h. After the reaction is completed, concentrate and purify by column chromatography to obtain the title compound.

Step 2 Preparation of 4-bromo-5-bromomethylthiazole

Weigh 5g of the obtained 4-bromo-5-methylthiazole in step 1 and 5g of NBS into a 250mL single-necked bottle, add 80mL of carbon tetrachloride to dissolve, reflux for 2h, after the reaction is completed, cool to room temperature, concentrate, column layer The title compound was obtained by analytical purification.

Step 3 Preparation of 4-bromo-5-hydroxymethylthiazole

Weigh 2g of the obtained 4-bromo-5-bromomethylthiazole in step 2, 3.2g of potassium carbonate in a 100ml single-neck bottle, add 50mL of 1,4-dioxane and 20mL of water to dissolve, react at 90°C for 1h, the reaction After completion, concentration and column chromatography were performed to obtain the title compound.

Step 4 Preparation of 4-bromo-5-formylthiazole

Weigh 1 g of 4-bromo-5-hydroxymethylthiazole obtained in step 3 into a 100 mL single-neck flask, add 30 mL of dichloromethane to dissolve, add 1.5 g of pyridine chlorochromate (PCC) in batches, complete the addition, and react at room temperature for 3 h , after the reaction was completed, concentrated and purified by column chromatography to obtain the title compound.

Step 5 Preparation of 4-(5-formylthiazol-4-yl)phenol

Using 4-bromo-5-formylthiazole obtained in step 4 and 4-hydroxyphenylboronic acid as raw materials, the title compound was obtained by the same method as step 1 in Example 9.

Step 6 4-(thiazol-5-carbaldehyde oxime-4-yl)phenol

Weigh 1 g of 4-(5-formylthiazol-4-yl)phenol obtained in step 5, 0.51 g of hydroxylamine hydrochloride and 0.6 g of sodium acetate into a 100-mL single-neck flask, add 30 mL of ethanol and 5 mL of water to dissolve, and reflux for 1 h. After the reaction was completed, it was cooled to room temperature, concentrated, and purified by column chromatography to obtain the title compound.

Step 7 Preparation of 4-(5-cyanothiazol-4-yl)phenol

Weigh 1g of 4-(thiazole-5-carbaldehyde oxime-4-yl)phenol obtained in step 6 into a 100ml single-necked flask, add 10mL of THF to dissolve, add 8mL of pyridine and 8mL of trifluoroacetic anhydride, and react at room temperature for 12h, the reaction ends After that, it was cooled to room temperature, concentrated, and purified by column chromatography to obtain the title compound.

Step 8(2S)-2-(((4-(5-Cyanothiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate preparation

4-(5-Cyanothiazol-4-yl)phenol obtained in step 7, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'- Deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.49(s, 1H), 8.74(d, 1H), 8.05(d, 2H), 7.02(d, 1H), 7.54(d, 1H), 7.41( d,2H),6.22(m,1H),6.17(m,1H),5.85(m,1H),4.88-4.81(m,1H),4.35(m,1H),4.22(m,1H),4.043 .78(m,1H), 3.89-3.31(m,2H), 1.29-1.22(m,6H), 1.16-1.14(m,6H).

LC-MS m/z: [M+H] ⁺ =638.

Example 17(2S)-2-(((3-methylbenzo[d]isoxazol-6-yloxy)(((2R,3R,4R,5R)-5-(2,4- Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)isopropionic acid Propyl ester

Step 1 (Z)-Preparation of 1-(2,4-dihydroxyphenyl)ethanone oxime

Weigh 5g of 1-(2,4-dihydroxyphenyl)ethanone into a 100mL single-necked bottle, add 50mL of ethanol to dissolve, add 2.3g of hydroxylamine hydrochloride, 2.7g of sodium acetate and 5mL of water, reflux for 2h, and the reaction ends After that, it was concentrated and purified by column chromatography to obtain the title compound.

Step 2 Preparation of 6-hydroxy-3-methylbenzo[d]isoxazole

Weigh 1 g of dichlorodicyanobenzoquinone (DDQ) and 1.2 g of PPh ₃ in a 100 mL single-necked bottle, add 30 mL of dichloromethane, stir to dissolve, and add dropwise 30 mL to dissolve 0.5 g of the result of step 1 (Z)-1 The solution of -(2,4-dihydroxyphenyl)ethanone oxime in dichloromethane was reacted at room temperature for 0.5 h, after the reaction was completed, concentrated and purified by column chromatography to obtain the title compound.

Step 3(2S)-2-(((3-methylbenzo[d]isoxazol-6-yloxy)(((2R,3R,4R,5R)-5-(2,4-di Isopropyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate Preparation of esters

6-Hydroxy-3-methylbenzo[d]isoxazole, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2 obtained in step 2 '-Deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d ₆ ) δppm: 11.49(s, 1H), 7.63(d, 1H), 7.53(d, 2H), 7.19(d, 1H), 6.10-6.07(m, 2H), 5.84(m,1H),5.54(m,1H),4.88-4.81(m,1H),4.35(m,1H),4.22(m,1H),4.043.78(m,1H),3.89-3.31( m, 2H), 2.56 (s, 3H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =585.

Example 18(2S)-2-(((4-(5-chlorothiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

Step 1 Preparation of 4-(thiazol-4-yl)phenol

Using 4-bromothiazole and 4-hydroxybenzeneboronic acid as starting materials, the title compound was obtained by the same method as in Example 1, step 1.

LC-MS m/z: [M+H] ⁺ =178.

Step 2 Preparation of 4-(5-chlorothiazol-4-yl)phenol

Weigh 1 g of 4-(thiazol-4-yl)phenol obtained in step 1 into a 100 mL single-necked flask, add 50 mL of dichloromethane to dissolve, add 0.75 g of NCS, complete the addition, and react at room temperature for 12 h. After the reaction is completed, concentrate, Column chromatography gave the title compound.

Step 3(2S)-2-(((4-(5-chlorothiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)isopropyl propionate preparation

4-(5-Chlorothiazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxygenate obtained in step 2 -2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.51(s, 1H), 9.13(s, 1H), 7.92(d, 2H), 7.57(d, 1H), 7.35(d, 2H), 6.17- 6.07(m, 2H), 5.85(m, 1H), 5.56(m, 1H) 4.88-4.81(m, 1H), 4.35(m, 1H), 4.22(m, 1H), 4.043.78(m, 1H) ), 3.89-3.31 (m, 2H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =647.

Example 19(2S)-2-(((4-(3,5-dimethylisoxazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-( 2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino ) isopropyl propionate

Step 1 Preparation of 4-(3,5-dimethylisoxazol-4-yl)phenol

Using 4-bromo-3,5-dimethylisoxazole and p-hydroxyphenylboronic acid as starting materials, the title compound was obtained by the same method as Example 1, step 1.

Step 2(2S)-2-(((4-(3,5-Dimethylisoxazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2 ,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino) isopropyl propionate

4-(3,5-Dimethylisoxazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R) obtained in step 1 )-2'-deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same methods as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d ₆ ) δppm: 11.49(s, 1H), 7.57(s, 1H), 7.39(d, 2H), 7.31(d, 2H), 6.00-6.11(m, 2H), 5.85(s, 1H), 5.51(m, 1H), 4.83-4.89(m, 1H), 4.39(m, 1H), 4.27(m, 1H), 4.02(m, 1H), 3.83-3.87(m, 2H), 2.38(s, 3H), 2.21(s, 3H), 1.23-1.28(m, 6H), 1.14-1.16(m, 6H).

LC-MS m/z: [M+H] ⁺ =625.

Example 20(2S)-2-(((4-(4-Chloro-1-methyl-1H-pyrazol-3-yl)phenyloxy)(((2R,3R,4R,5R)- 5-(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphorus Acyl)amino)isopropyl propionate

Step 1 4-(4-Chloro-1-methyl-1H-pyrazol-3-yl)phenol

Weigh 0.5 g of 4-(1-methyl-1H-pyrazol-3-yl)phenol (2.9 mmol) and 0.385 g of NCS (2.9 mmol) prepared in step 1 of Example 15 into a 50 mL eggplant-shaped flask, add 5 mL THF and 5 mL of CH ₃ CN were reacted at 50°C for 3 h. After the reaction was completed, 100 mL of ethyl acetate and 50 mL of saturated aqueous sodium chloride were added for extraction, washed three times with water, and then the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated. Column chromatography gave the title compound.

LC-MS m/z: [M+H] ⁺ =209.

Step 2(2S)-2-(((4-(4-Chloro-1-methyl-1H-pyrazol-3-yl)phenyloxy)(((2R,3R,4R,5R)-5 -(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl ) preparation of amino) isopropyl propionate

Using the resultant of step 1, 4-(4-chloro-1-methyl-1H-pyrazol-3-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and ( 2'R)-2'-deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.49(s, 1H), 8.02(s, 1H), 7.78-7.80(m, 2H), 7.56-7.57(m, 1H), 7.29-7.31(m ,2H),6.00-6.09(m,2H),5.82-5.84(m,1H),5.57-5.58(m,1H),4.83-4.88(m,1H),4.37-4.40(m,1H),4.24 -4.26(m, 1H), 4.00-4.03(m, 1H), 3.80-3.86(m, 5H), 1.23-1.29(m, 6H), 1.14-1.15(m, 6H).

LC-MS m/z: [M+H] ⁺ =644.

Example 21(2S)-2-(((4-(5-methyloxazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4- Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)isopropionic acid Propyl ester

Step 1 Preparation of 2-bromo-1-(4-methoxyphenyl)-1-propanone

Weigh 1g 1-(4-methoxyphenyl)-1-propanone (6.1mmol), 1.2g p-toluenesulfonic acid monohydrate (6.1mmol) and 1.1g NBS (6.1mmol) in a 100mL single-neck flask , 30 mL of acetonitrile was added to dissolve, and the reaction was refluxed for 4 h. After the reaction was completed, the mixture was concentrated and purified by column chromatography to obtain the title compound.

Step 2 Preparation of 4-(4-methoxyphenyl)-5-methyloxazole

Weigh 1 g of 2-bromo-1-(4-methoxyphenyl)-1-propanone (4.4 mmol) obtained in step 1 into a 100 mL single-neck flask, add 2 mL of formamide and 20 mL of DMF, and react under reflux for 2 h. After the reaction, concentrated, poured the concentrated solution into ice water, extracted with ethyl acetate, dried, filtered, concentrated, and purified by column chromatography to obtain the title compound.

Step 3 Preparation of 4-(5-methyloxazol-4-yl)phenol

Weigh 0.4 g of the obtained 4-(4-methoxyphenyl)-5-methyloxazole (1.7 mmol) in step 2 into a 100 mL single-neck flask, add 20 mL of dichloromethane to dissolve, and add 8 mL dropwise under an ice bath Dissolve 8 mmol of boron tribromide in dichloromethane solution, complete the addition, and react at room temperature for 12 h. After the completion of the raw materials, add water under ice bath to quench, extract with dichloromethane, dry, filter, concentrate, and purify by column chromatography to obtain the title compound.

Step 4(2S)-2-(((4-(5-methyloxazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Isopropyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate Preparation of esters

4-(5-Methyloxazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2' obtained in step 3 -Deoxy-2'-fluoro-2'-methyluridine is used as the raw material, and the target compound is obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.51(s, 1H), 8.28(s, 1H), 7.68(d, 2H), 7.56(d, 1H), 7.31(d, 2H), 6.13- 6.05(m, 2H), 5.87(m, 1H), 5.56(m, 1H), 4.88-4.81(m, 1H), 4.35(m, 1H), 4.22(m, 1H), 4.04-3.78(m, 1H), 3.89-3.31 (m, 2H), 2.50 (s, 3H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =611.

Example 22 (2S)-2-(((1-methyl-1H-indol-4-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl ester

Step 1N-Methyl-4-methoxyindole

Weigh 1.47g of 4-methoxyindole (10mmol), 1.38g of potassium carbonate (10mmol) in a 250mL eggplant-shaped flask, add 100mL of acetone to dissolve, slowly add 2.84g of methyl iodide (20mmol), and react at room temperature. After the completion, 100 mL of ethyl acetate and 50 mL of saturated aqueous sodium chloride solution were added to the reaction solution for extraction, washed three times with water, and then the organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound.

LC-MS m/z: [M+H] ⁺ =162.

Step 2N-Methyl-4-hydroxyindole

Using N-methyl-4-methoxyindole obtained in step 1 as a starting material, the title compound was prepared in the same manner as in step 2 of Example 14.

LC-MS m/z: [M+H] ⁺ =148.

Step 3(2S)-2-(((1-Methyl-1H-indol-4-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo- Preparation of isopropyl 3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

N-methyl-4-hydroxyindole obtained in step 2, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy-2' -Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.47(s,1H), 7.46-7.48(m,1H), 7.31-7.32(m,1H), 7.25-7.27(m,1H), 7.05-7.11 (m,1H),6.95-6.97(m,1H),6.49-6.50(m,1H),5.96-6.04(m,2H),5.83-5.85(m,1H),5.33-5.35(m,1H) ,4.82-4.90(m,1H),4.35-4.41(m,1H),4.20-4.28(m,1H),3.88-4.00(m,1H),3.84-3.87(m,2H),3.78-3.82( m, 3H), 1.20-1.27 (m, 6H), 1.14-1.16 (m, 6H).

LC-MS m/z: [M+H] ⁺ =583.

Example 23 (2S)-2-(((4-(oxazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo- Isopropyl 3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

Step 1 Preparation of 2-bromo-1-(4-methoxyphenyl)ethanone

Weigh 1g 1-(4-methoxyphenyl)ethanone (6.7mmol), 1.5g p-toluenesulfonic acid monohydrate (6.7mmol) and 1.2g NBS (6.7mmol) in a 100mL single-neck flask, add 30 mL of acetonitrile was dissolved, and the reaction was refluxed for 4 hours. After the reaction was completed, it was concentrated and purified by column chromatography to obtain the title compound.

Step 2 Preparation of 4-(4-methoxyphenyl)oxazole

Weigh 1g of step 1 gained 2-bromo-1-(4-methoxyphenyl)ethanone (4.4mmol) in a 100mL single-necked flask, add 2mL formamide and 20mL DMF, reflux for 2h, after the raw material finishes , concentrated, the reaction solution was poured into ice water, extracted with ethyl acetate, dried, filtered, concentrated, and purified by column chromatography to obtain the title compound.

Step 3 Preparation of 4-(oxazol-4-yl)phenol

Weigh 0.4 g of step 2 obtained 4-(4-methoxyphenyl) oxazole (2.3 mmol) in a 100 mL single-necked flask, add 20 mL of dichloromethane to dissolve, and add 8 mL of boron tribromide under an ice bath. Dichloromethane solution (1.0 mol/L, 8 mL) was added and reacted at room temperature for 24 h. After the reaction, water was added under ice bath to quench, extracted with dichloromethane, dried, filtered, concentrated, and purified by column chromatography to obtain the title compound.

Step 4(2S)-2-(((4-(oxazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3 Preparation of isopropyl ,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

4-(oxazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy-2 obtained in step 3 '-Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same methods as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d ₆ ) δppm: 11.51(s, 1H), 8.59(s, 1H), 8.45(s, 1H), 7.78(d, 2H), 7.56(d, 1H), 7.31( d,2H),6.13-6.05(m,2H),5.87(m,1H),5.56(m,1H),4.88-4.81(m,1H),4.35(m,1H),4.22(m,1H) , 4.04-3.78 (m, 1H), 3.89-3.31 (m, 2H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =597.

Example 24 (2S)-2-(((4-(5-fluorothiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

Step 1 Preparation of 4-(5-Fluoro-thiazol-4-yl)phenol

Weigh 3 g of 4-(thiazol-4-yl)phenol (16 mmol) obtained in step 1 of Example 18, 6.0 g of 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octane Alkane bis(tetrafluoroborate) salt (16 mmol) was placed in a 100 mL single-necked flask, 50 mL of 1,4-dioxane was added to dissolve, and the reaction was refluxed for 12 h. After the reaction was completed, concentrated and purified by column chromatography to obtain the title compound.

Step 2(2S)-2-(((4-(5-fluorothiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)isopropyl propionate preparation

4-(5-Fluoro-thiazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'- Deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d ₆ ) δppm: 11.51(s, 1H), 8.75(s, 1H), 7.84(d, 2H), 7.56(d, 1H), 7.35(d, 2H), 6.17- 6.05(m, 2H), 5.86(m, 1H), 5.56(m, 1H), 4.88-4.81(m, 1H), 4.35(m, 1H), 4.22(m, 1H), 4.04-3.78(m, 1H), 3.89-3.31 (m, 2H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =631.

Example 25(2S)-2-(((2-Fluoro-4-(thiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Isopropyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate ester

Step 1 Preparation of 2-fluoro-4-(thiazol-4-yl)phenol

Weigh 3g of 4-(thiazol-4-yl)phenol (0.017mol), 6.0g of 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octanebis(tetrafluoroboric acid) ) salt (Selectfluor, 0.017mol) in a 250mL single-neck flask, add 30mL trifluoroacetic acid, react at 95°C for 3h, after the reaction is complete, concentrate, add water and ethyl acetate for extraction, dry, filter, concentrate, and purify by column chromatography to obtain title compound.

Step 2(2S)-2-(((2-Fluoro-4-(thiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate preparation

2-Fluoro-4-(thiazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'- Deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d ₆ ) δppm: 11.50(s, 1H), 9.21(s, 1H), 8.25(s, 1H), 7.96-7.81(m, 2H), 7.66-7.56(m, 2H) ),6.13-6.05(m,2H),5.87(m,1H),5.56(m,1H),4.88-4.81(m,1H),4.35(m,1H),4.22(m,1H),4.04- 3.78 (m, 1H), 3.89-3.75 (m, 2H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =631.

Example 26(2S)-2-(((4-(thiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3 ,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate isopropyl ester

Step 1 Preparation of 4-(thiophen-2-yl)phenol

Take by weighing 3g 4-hydroxyphenylboronic acid (22mmol), 3g 2-bromo-thiophene (18mmol), 7.5g potassium carbonate (55mmol), 1.2g PdCl ₂ (PPh _{) 2 (} 1.7mmol) in the single-necked flask of 250mL, Add 50 mL of 1,4-dioxane and 10 mL of water, react at 95°C for 3 h under nitrogen protection, after the reaction, concentrate, add water and ethyl acetate for extraction, dry, filter, concentrate, and purify by column chromatography to obtain the title compound.

Step 2(2S)-2-(((4-(thiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3, Preparation of isopropyl 4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

4-(thiophen-2-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy-2' obtained in step 1 -Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d ₆ ) δppm: 11.50 (s, 1H), 7.66-7.56 (m, 3H), 7.42-7.22 (m, 2H), 7.26 (d, 2H), 7.12 (m, 1H) ),6.13-6.05(m,2H),5.87(m,1H),5.56(m,1H),4.88-4.81(m,1H),4.35(m,1H),4.22(m,1H),4.04- 3.78 (m, 1H), 3.89-3.75 (m, 2H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =612.

Example 27 (2S)-2-(((3-fluoro-4-(thiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Isopropyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate ester

Step 1 Preparation of 3-fluoro-4-(thiazol-4-yl)phenol

Using 4-hydroxy-2-fluorophenylboronic acid and 4-bromo-thiazole as starting materials, the title compound was obtained by the same method as step 1 of Example 26.

Step 2(2S)-2-(((3-Fluoro-4-(thiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate preparation

3-Fluoro-4-(thiazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'- Deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.49(s, 1H), 9.23(s, 1H), 8.17(m, 1H), 8.13(s, 1H), 7.55(d, 1H), 7.30- 7.19(m, 2H), 6.13-6.05(m, 2H), 5.87(m, 1H), 5.56(m, 1H), 4.88-4.81(m, 1H), 4.35(m, 1H), 4.22(m, 1H), 4.04-3.78 (m, 1H), 3.89-3.75 (m, 2H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =631.

Example 28(2S)-2-(((3-methyl-4-(thiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4- Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)isopropionic acid Propyl ester

Step 1 Preparation of 3-methyl-4-(thiazol-4-yl)phenol

Using 4-hydroxy-2-methylphenylboronic acid and 4-bromothiazole as starting materials, the title compound was prepared in the same manner as in Example 26, step 1.

Step 2(2S)-2-(((3-methyl-4-(thiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Isopropyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate ester

3-methyl-4-(thiazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2' obtained in step 1 -Deoxy-2'-fluoro-2'-methyluridine is used as the raw material, and the target compound is obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d ₆ ) δppm: 11.51(s, 1H), 9.19(s, 1H), 7.80(s, 1H), 7.60(d, 2H), 7.17(m, 2H), 6.13- 6.05(m, 2H), 5.87(m, 1H), 5.56(m, 1H), 4.88-4.81(m, 1H), 4.35(m, 1H), 4.22(m, 1H), 4.04-3.78(m, 1H), 3.89-3.75 (m, 2H), 2.39 (s, 3H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =627.

Example 29(2S)-2-(((3-methyl-4-(1-methyl-1H-imidazol-4-yl)phenyloxy)(((2R,3R,4R,5R)- 5-(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphorus Acyl)amino)isopropyl propionate

Step 1 Preparation of 3-methyl-4-(1-methyl-1H-imidazol-4-yl)phenol

Using 4-hydroxy-2-methylbenzeneboronic acid and 4-bromo-N-methylimidazole as starting materials, the title compound was obtained by the same method as in Example 26, step 1.

Step 2(2S)-2-(((3-Methyl-4-(1-methyl-1H-imidazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5 -(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl ) amino) isopropyl propionate

Using the resultant of step 1, 3-methyl-4-(1-methyl-1H-imidazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and ( 2'R)-2'-deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.51 (s, 1H), 7.80-7.31 (m, 4H), 7.07 (m, 2H), 6.13-6.05 (m, 2H), 5.87 (m, 1H) ), 5.56(m, 1H), 4.88-4.81(m, 1H), 4.35(m, 1H), 4.22(m, 1H), 4.04-3.78(m, 1H), 3.89-3.75(m, 2H), 3.71(s.3H), 3.54(s,3H), 1.29-1.22(m,6H), 1.16-1.14(m,6H).

LC-MS m/z: [M+H] ⁺ =624.

Example 30(2S)-2-(((4-(1-methyl-1H-imidazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2, 4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propane isopropyl acid

Step 1 Preparation of 4-(1-methyl-1H-imidazol-4-yl)phenol

Using 4-hydroxyphenylboronic acid and 4-bromo-N-methylimidazole as starting materials, the title compound was prepared in the same manner as in Example 26, step 1.

Step 2(2S)-2-(((4-(1-methyl-1H-imidazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4 -Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionic acid Preparation of isopropyl ester

4-(1-Methyl-1H-imidazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl hydrochloride, pentafluorophenol and (2'R)- 2'-Deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d ₆ ) δppm: 11.510 (s, 1H), 7.72-7.58 (m, 5H), 7.20 (m, 2H), 6.04-6.00 (m, 2H), 5.89 (m, 1H) ), 5.58(m, 1H), 4.88-4.81(m, 1H), 4.35(m, 1H), 4.22(m, 1H), 4.043.78(m, 1H), 3.89-3.31(m, 2H), 3.68(s, 3H), 1.28-1.22(m, 6H), 1.16-1.14(m, 6H).

LC-MS m/z: [M+H] ⁺ =610.

Example 31 (2S)-2-(((4-(3-chloropyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

Step 1 Preparation of 3-chloro-2-(4-hydroxyphenyl)pyridine

Weigh 1.38g of 4-hydroxyphenylboronic acid (10mmol), 1.91g of 2-bromo-3-chloropyridine (10mmol), 0.4g of [1,1'-bis(diphenylphosphonium)ferrocene]palladium dichloride (0.5mmol), 9g of cesium carbonate (30mmol) was placed in a 250mL eggplant-shaped flask, 50mL of 1,4-dioxane and 5mL of water were added, and the reaction was carried out at 90°C for 1.5h under nitrogen protection. After the reaction, 100mL of ethyl acetate was added. The ester was extracted with 50 mL of saturated aqueous sodium chloride solution, washed three times with water, and then the organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound.

LC-MS m/z: [M+H] ⁺ =206.

Step 2(2S)-2-(((4-(3-chloropyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)isopropyl propionate preparation

3-Chloro-2-(4-hydroxyphenyl)pyridine, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'- Deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.47 (s, 1H), 8.61-8.62 (m, 1H), 8.02-8.05 (m, 1H), 7.69-7.72 (m, 2H), 7.56-7.59 (m,1H),7.41-7.45(m,1H),7.32-7.35(m,2H),6.14-6.17(m,1H),6.05-6.09(m,1H),5.84-5.98(m,1H) ,5.57-5.60(m,1H),4.83-4.91(m,1H),4.38-4.43(m,1H),4.25-4.29(m,1H),4.01-4.05(m,1H),3.80-3.89( m, 2H), 1.22-1.29 (m, 6H), 1.15-1.17 (m, 6H).

LC-MS m/z: [M+H] ⁺ =641.

Example 32(2S)-2-(((4-(5-chloropyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

Step 1 Preparation of 4-(5-chloropyridin-2-yl)phenol

Using 4-hydroxyphenylboronic acid and 2-bromo-5-chloropyridine as starting materials, the title compound was prepared by the same method as in Example 31, step 1.

LC-MS m/z: [M+H] ⁺ =206.

Step 2(2S)-2-(((4-(5-chloropyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo -3,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)isopropyl propionate preparation

4-(5-Chloropyridin-2-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxygenate obtained in step 1 -2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.47(s, 1H), 8.68(m, 1H), 8.08-8.11(m, 1H), 7.82-7.85(m, 1H), 7.55-7.59(m ,1H),7.48-7.53(m,1H),7.31-7.35(m,2H),7.15-7.20(m,1H),6.07-6.14(m,2H),5.99(m,1H),5.57-5.60 (m,1H),4.85-4.89(m,1H),4.37-4.41(m,1H),4.26-4.28(m,1H),4.01(m,1H),3.80-3.87(m,2H),1.22 -1.29(m, 6H), 1.13-1.16(m, 6H).

LC-MS m/z: [M+H] ⁺ =641.

Example 33(2S)-2-(((4-(5-chloropyridin-2-yl)-3-methylphenyloxy)(((2R,3R,4R,5R)-5-(2 ,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino) isopropyl propionate

Step 1 Preparation of 4-(5-chloropyridin-2-yl)-3-methylphenol

Using 2-methyl-4-hydroxybenzeneboronic acid and 2-bromo-5-chloropyridine as starting materials, the title compound was obtained by the same method as Example 31, step 1.

LC-MS m/z: [M+H] ⁺ =220.

Step 2(2S)-2-(((4-(5-chloropyridin-2-yl)-3-methylphenyloxy)(((2R,3R,4R,5R)-5-(2, 4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propane isopropyl acid

4-(5-Chloropyridin-2-yl)-3-methylphenol obtained in step 1, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R) -2'-Deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.51 (s, 1H), 8.69 (m, 1H), 7.54-7.57 (m, 2H), 7.39-7.42 (m, 1H), 7.14-7.17 (m ,2H),6.05-6.12(m,2H),5.85(m,1H),5.54-5.57(m,1H),4.82-4.91(m,1H),4.36-4.42(m,1H),4.21-4.27 (m, 2H), 3.90-4.01 (m, 1H), 3.82-3.88 (m, 2H), 2.30 (m, 3H), 1.21-1.29 (m, 6H), 1.14-1.16 (m, 6H).

LC-MS m/z: [M+H] ⁺ =655.

Example 34 (2S)-2-(((4-(3-fluoropyridin-2-yl)-3-methylphenyloxy)(((2R,3R,4R,5R)-5-(2 ,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino) isopropyl propionate

Step 1 Preparation of 4-(3-fluoropyridin-2-yl)-3-methylphenol

Using 2-methyl-4-hydroxybenzeneboronic acid and 2-bromo-3-fluoropyridine as starting materials, the title compound was obtained by the same method as Example 31, step 1.

LC-MS m/z: [M+H] ⁺ =204.

Step 2(2S)-2-(((4-(3-Fluoropyridin-2-yl)-3-methylphenyloxy)(((2R,3R,4R,5R)-5-(2, 4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propane isopropyl acid

4-(3-Fluoropyridin-2-yl)-3-methylphenol obtained in step 1, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R) -2'-Deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.51 (s, 1H), 8.53 (m, 1H), 8.13 (m, 1H), 7.79-7.86 (m, 1H), 7.50-7.57 (m, 1H) ),7.31-7.34(m,1H),7.15-7.20(m,2H),6.06-6.13(m,2H),5.84(m,1H),5.58-5.60(m,1H),4.85-4.89(m ,1H),4.39(m,1H),4.28(m,1H),4.03(m,1H),3.83-3.85(m,2H),2.15(m,3H),1.22-1.29(m,6H), 1.15-1.17 (m, 6H).

LC-MS m/z: [M+H] ⁺ =639.

Example 35 (2S)-2-(((4-(5-chlorooxazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Isopropyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate ester

Step 1 Preparation of 2-bromo-1-(4-methoxyphenyl)ethanone

Using 1-(4-methoxyphenyl)ethanone as starting material, the title compound was prepared in the same manner as in Example 21, step 1.

Step 2 Preparation of 4-(4-methoxyphenyl)oxazole

Using 2-bromo-1-(4-methoxyphenyl)ethanone obtained in step 1 and formamide as starting materials, the title compound was prepared in the same manner as in step 2 of Example 21.

LC-MS m/z: [M+H] ⁺ =176.

Step 3 Preparation of 5-chloro-4-(4-methoxyphenyl)oxazole

Weigh 3.0 g of 4-(4-methoxyphenyl) oxazole (0.0171 mol) obtained in step 2 into a 100 ml single-neck flask, add 30 mL of acetonitrile to dissolve, add 2.29 g of NCS (0.0171 mol), and react at room temperature for 2 h , after the reaction was completed, concentrated and purified by column chromatography to obtain the title compound.

LC-MS m/z: [M+H] ⁺ =210.

Step 4 Preparation of 4-(5-chlorooxazol-4-yl)phenol

Weigh 1.3g of step 3 obtained 5-chloro-4-(4-methoxyphenyl)oxazole (6.22mmol) in a 50mL single-neck flask, add 15mL of methanesulfonic acid to dissolve, add 3.71g of DL-Met (24.8 mmol), the addition was completed, and the reaction was carried out at 60 °C for 24 h. After the reaction, cooled, poured into ice water, extracted with ethyl acetate (50 mL x 6), dried, filtered, concentrated, and purified by column chromatography to obtain the title compound.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ ppm: 9.76 (s, 1H), 8.48 (s, 1H), 7.69 (d, 2H), 6.88 (d, 2H).

LC-MS m/z: [M+H] ⁺ =196.

Step 5(2S)-2-(((4-(5-chlorooxazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate preparation

4-(5-Chloroxazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'- Deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d ₆ ) δppm: 11.50(s, 1H), 8.55(s, 1H), 7.87(d, 2H), 7.56(d, 1H), 7.37(d, 2H), 5.90- 6.20(m, 2H), 5.85(d, 1H), 5.57(d, 1H), 4.77-4.92(m, 1H), 4.20-4.45(m, 2H), 3.95-4.10(m, 1H), 3.70- 3.95(m, 2H), 1.18-1.40(m, 6H), 1.15(d, 6H).

LC-MS m/z: [M+H] ⁺ =631.

Example 36 (2S)-2-(((4-(3-chloropyridin-2-yl)-3-methylphenyloxy)(((2R,3R,4R,5R)-5-(2 ,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino) isopropyl propionate

Step 1 Preparation of 4-(3-chloropyridin-2-yl)-3-methylphenol

Using 2-bromo-3-chloropyridine and 4-hydroxy-2-methylbenzeneboronic acid as starting materials, the title compound was obtained by the same method as Example 1, step 1.

LC-MS m/z: [M+H] ⁺ =220.

Step 2(2S)-2-(((4-(3-chloropyridin-2-yl)-3-methylphenyloxy)(((2R,3R,4R,5R)-5-(2, 4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propane Preparation of isopropyl acid

4-(3-Chloropyridin-2-yl)-3-methylphenol obtained in step 1, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R) -2'-Deoxy-2'-fluoro-2'-methyluridine was used as the starting material, and the title compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ HNMR (300MHz, DMSO-d6) δppm: 11.48(s, 1H), 8.61(d, 1H), 8.01(d, 1H), 7.58(d, 1H), 7.46(d, 1H), 7.23-7.02( m,3H),6.20-5.90(m,3H),5.58(m,1H),4.88-4.83(m,1H),4.39-4.26(m,2H),4.01-3.98(m,1H),3.90- 3.78(m, 2H), 2.04(s, 3H), 1.29-1.22(m, 6H), 1.16-1.14(m, 6H).

LC-MS m/z: [M+H] ⁺ =655. .

Example 37(2S)-2-(((4-(5-Chloro-1-methyl-1H-imidazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5 -(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl ) amino) isopropyl propionate

Step 1 Preparation of 4-(5-chloro-1-methyl-1H-imidazol-4-yl)phenol

Weigh 0.81g of Example 30 step 1 obtained 4-(1-methyl-1H-imidazol-4-yl)phenol (5.14mmol), 0.62g NCS (5.14mmol) in a 100mL single-neck flask, add 20mL of acetonitrile Dissolved, refluxed for 1 h, after the reaction, concentrated and purified by column chromatography to obtain the title compound.

Step 2(2S)-2-(((4-(5-Chloro-1-methyl-1H-imidazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5- (2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl) Preparation of isopropyl amino)propionate

4-(5-Chloro-1-methyl-1H-imidazol-4-yl)phenol obtained in step 1, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2 'R)-2'-deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as in steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d ₆ ) δppm: 11.51(s, 1H), 8.40(s, 1H), 7.84(d, 2H), 7.56(d, 1H), 7.35(d, 2H), 6.17- 6.05(m, 2H), 5.86(m, 1H), 5.56(m, 1H), 4.88-4.81(m, 1H), 4.35(m, 1H), 4.22(m, 1H), 4.043.78(m, 1H), 3.89-3.31 (m, 2H), 3.62 (s, 3H), 1.29-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =644.

Example 38(2S)-2-(((3-Fluoro-4-(1-methyl-1H-imidazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5 -(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl ) amino) isopropyl propionate

Step 1 3-Fluoro-4-(1-methyl-1H-imidazol-4-yl)phenol

Using 4-bromo-1-methyl-1H-imidazole and 2-fluoro-4-hydroxybenzeneboronic acid as starting materials, the title compound was obtained by the same method as step 1 of Example 26.

Step 2(2S)-2-(((3-Fluoro-4-(1-methyl-1H-imidazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5- (2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl) Preparation of isopropyl amino)propionate

3-fluoro-4-(1-methyl-1H-imidazol-4-yl)phenol obtained in step 1, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2 'R)-2'-deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as in steps 2 and 3 of Example 1.

LC-MS m/z: [M+H] ⁺ =628.

Example 39 (2S)-2-(((4-(2-methylthiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Isopropyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate ester

Step 1 Preparation of 4-(2-methylthiazol-4-yl)phenol

Using 2-methyl-4-bromothiazole and 4-hydroxybenzeneboronic acid as starting materials, the title compound was obtained by the same method as in Example 1, step 1.

LC-MS m/z: [M+H] ⁺ =192.

Step 2(2S)-2-(((4-(2-Methylthiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate preparation

4-(2-Methylthiazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'- Deoxy-2'-fluoro-2'-methyluridine was used as a raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

LC-MS m/z: [M+H] ⁺ =627.

Example 40(2S)-2-(((4-(5-Chloro-2-methylthiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2 ,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino) isopropyl propionate

Step 1 Preparation of 4-(5-chloro-2-methylthiazol-4-yl)phenol

Using 4-(2-methylthiazol-4-yl)phenol obtained in step 1 of Example 39 as a starting material, the title compound was prepared in the same manner as in step 1 of Example 37.

LC-MS m/z: [M+H] ⁺ =226.

Step 2(2S)-2-(((4-(5-Chloro-2-methylthiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2, 4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propane Preparation of isopropyl acid

4-(5-Chloro-2-methylthiazol-4-yl)phenol, phosphorus oxychloride, L-alanine isopropyl hydrochloride, pentafluorophenol and (2'R) obtained in step 1 -2'-Deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

LC-MS m/z: [M+H] ⁺ =661.

Example 41 (2S)-2-(((4-(5-Chloro-2-methylthiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2 ,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino) methyl propionate

4-(5-Chloro-2-methylthiazol-4-yl)phenol, phosphorus oxychloride, L-alanine methyl ester hydrochloride, pentafluorophenol and (2' R)-2'-deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

LC-MS m/z: [M+H] ⁺ =633.

Example 42 (2S)-2-(((4-(5-Chloro-thiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Methyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

4-(5-Chlorothiazol-4-yl)phenol, phosphorus oxychloride, L-alanine methyl ester hydrochloride, pentafluorophenol and (2'R)-2' obtained in step 2 of Example 18 -Deoxy-2'-fluoro-2'-methyluridine is used as the raw material, and the target compound is obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d6) δppm: 11.50(s, 1H), 9.13(s, 1H), 7.92(d, 2H), 7.56(d, 1H), 7.35(d, 2H), 6.20-6.02 (m,2H),5.87(m,1H),5.58(m,1H),4.39-4.03(m,2H),4.01-3.98(m,1H),3.93-3.85(m,2H),3.58(s , 3H), 1.28-1.22 (m, 6H).

LC-MS m/z: [M+H] ⁺ =619.

Example 43 (2S)-2-(((4-(5-chlorothiazol-4-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Ethyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

4-(5-Chlorothiazol-4-yl)phenol, phosphorus oxychloride, L-alanine ethyl ester hydrochloride, pentafluorophenol and (2'R)-2' obtained in step 2 of Example 18 -Deoxy-2'-fluoro-2'-methyluridine is used as the raw material, and the target compound is obtained by the same method as steps 2 and 3 of Example 1.

LC-MS m/z: [M+H] ⁺ =633.

Example 44 (2S)-2-(((4-(3-fluoropyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Methyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate

4-(3-fluoropyridin-2-yl)phenol, phosphorus oxychloride, L-alanine methyl ester hydrochloride, pentafluorophenol and (2'R)-2' obtained in Example 2, step 1 -Deoxy-2'-fluoro-2'-methyluridine is used as the raw material, and the target compound is obtained by the same method as steps 2 and 3 of Example 1.

LC-MS m/z: [M+H] ⁺ =597.

Example 45 (2S)-2-(((4-(3-fluoropyridin-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Ethyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

4-(3-fluoropyridin-2-yl)phenol, phosphorus oxychloride, L-alanine ethyl ester hydrochloride, pentafluorophenol and (2'R)-2' obtained in Example 2, step 1 -Deoxy-2'-fluoro-2'-methyluridine is used as the raw material, and the target compound is obtained by the same method as steps 2 and 3 of Example 1.

LC-MS m/z: [M+H] ⁺ =611.

Example 46(2S)-2-(((Benzo[b]thiophen-4-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3, Isopropyl 4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

With 4-hydroxybenzothiophene, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy-2'-fluoro-2'-methyl Using uridine as a raw material, the target compound was prepared by the same methods as steps 2 and 3 in Example 1.

¹ H NMR (500MHz, DMSO-d6) δppm: 11.49 (s, 1H), 7.84-7.78 (m, 2H), 7.51 (d, 2H), 7.37-7.30 (m, 2H), 6.20-6.02 (m, 2H), 5.88(m, 1H), 5.43(m, 1H), 4.86(m, 1H), 4.43-4.03(m, 2H), 4.01-3.98(m, 1H), 3.93-3.85(m, 2H) , 1.28-1.22 (m, 6H), 1.14-1.12 (m, 6H).

LC-MS m/z: [M+H] ⁺ =586.

Example 47(2S)-2-(((Benzo[b]thiophen-4-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3, Methyl 4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate

With 4-hydroxybenzothiophene, phosphorus oxychloride, L-alanine methyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy-2'-fluoro-2'-methylurine Using glycosides as raw materials, the target compound was obtained by the same methods as steps 2 and 3 in Example 1.

LC-MS m/z: [M+H] ⁺ =558.

Example 48(2S)-2-(((3-methylbenzo[b]thiophen-4-yloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Isopropyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

With 3-methyl-4-hydroxybenzothiophene, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy-2'-fluoro- 2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 in Example 1.

LC-MS m/z: [M+H] ⁺ =600.

Example 49 (2S)-2-(((4-(3-methylthiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Methyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

Step 1 Preparation of 4-(3-methylthiophen-2-yl)phenol

Using 2-bromo-3-methylthiophene and 4-hydroxybenzeneboronic acid as starting materials, the title compound was prepared in the same manner as in Example 1, step 1.

LC-MS m/z: [M+H] ⁺ =191.

Step 2(2S)-2-(((4-(3-Methylthiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Methyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

4-(3-Methylthiophen-2-yl)phenol, phosphorus oxychloride, L-alanine methyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxygenated from step 1 -2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d6) δppm: 11.50(s, 1H), 7.57(s, 1H), 7.46-7.43(m, 3H), 7.30(d, 2H), 7.00(d, 1H), 6.20 -6.02(m, 2H), 5.87(m, 1H), 5.58(m, 1H), 4.39-4.03(m, 2H), 4.01-3.98(m, 1H), 3.93-3.85(m, 2H), 3.60 (s, 3H), 2.24 (s, 3H), 1.28-1.22 (m, 6H). LC-MS m/z: [M+H] ⁺ =598.

Example 50(2S)-2-(((4-(3-methylthiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Isopropyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate ester

4-(3-Methylthiophen-2-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)- 2'-Deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d6) δppm: 11.50(s, 1H), 7.57(s, 1H), 7.46-7.43(m, 3H), 7.30(d, 2H), 7.00(d, 1H), 6.20 -6.02(m, 2H), 5.87(m, 1H), 5.58(m, 1H), 4.88-4.83(m, 1H), 4.39-4.03(m, 2H), 4.01-3.98(m, 1H), 3.93 -3.85(m, 2H), 2.26(s, 3H), 1.28-1.22(m, 6H), 1.17-1.14(m, 6H).

LC-MS m/z: [M+H] ⁺ =626.

Example 51 (2S)-2-(((4-(5-methylthiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Methyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate

Step 1 Preparation of 4-(5-methylthiophen-2-yl)phenol

Using 2-bromo-5-methylthiophene and 4-hydroxybenzeneboronic acid as starting materials, the title compound was prepared in the same manner as in Example 1, step 1.

LC-MS m/z: [M+H] ⁺ =191.

Step 2(2S)-2-(((4-(5-Methylthiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo Methyl substituted-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate

4-(5-Methylthiophen-2-yl)phenol, phosphorus oxychloride, L-alanine methyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxygenated from step 1 -2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the method of Example 1.

¹ H NMR (300MHz, DMSO-d6) δppm: 11.51(s, 1H), 7.59-7.55(m, 3H), 7.25-7.21(m, 3H), 6.81(d, 1H), 6.20-6.02(m, 2H), 5.87(m, 1H), 5.58(m, 1H), 4.39-4.03(m, 2H), 4.01-3.98(m, 1H), 3.93-3.85(m, 2H), 3.30(s, 3H) , 2.46(s, 3H), 1.29-1.22(m, 6H).

LC-MS m/z: [M+H] ⁺ =598.

Example 52 (2S)-2-(((4-(5-methylthiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Isopropyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanoate Ester

4-(5-methylthiophen-2-yl)phenol, phosphorus oxychloride, L-alanine isopropyl ester hydrochloride, pentafluorophenol and (2'R)- 2'-Deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d6) δppm: 11.50(s, 1H), 7.59-7.55(m, 3H), 7.25-7.22(m, 3H), 6.80(d, 1H), 6.20-6.02(m, 2H), 5.87(m, 1H), 5.58(m, 1H), 4.88-4.83(m, 1H), 4.39-4.03(m, 2H), 4.01-3.98(m, 1H), 3.93-3.85(m, 2H), 2.46 (s, 3H), 1.28-1.22 (m, 6H), 1.16-1.14 (m, 6H).

LC-MS m/z: [M+H] ⁺ =626.

Example 53 (2S)-2-(((4-(5-methylthiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Ethyl oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

4-(5-methylthiophen-2-yl)phenol, phosphorus oxychloride, L-alanine ethyl ester hydrochloride, pentafluorophenol and (2'R)-2 obtained in Example 51, step 1 '-Deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d6) δppm: 11.50(s, 1H), 7.55-7.57(m, 3H), 7.21-7.25(m, 3H), 6.80-6.81(m, 1H), 6.60-6.61( m,1H),5.98-6.04(m,1H),5.83-5.86(m,1H),5.55-5.58(m,1H),4.34-4.40(m,1H),4.20-4.26(m,1H), 3.90-3.93 (m, 1H), 3.85-3.88 (m, 2H), 3.58 (m, 2H), 2.45-2.49 (m, 3H), 1.21-1.29 (m, 9H).

LC-MS m/z: [M+H] ⁺ =612.

Example 54 (2S)-2-(((4-(thiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3 ,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate ethyl ester

4-(thiophen-2-yl)phenol, phosphorus oxychloride, L-alanine ethyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy- 2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was prepared in the same manner as in steps 2 and 3 of Example 1.

¹ H NMR (300MHz, DMSO-d6) δppm: 11.50(s, 1H), 7.65(d, 2H), 7.58-7.52(m, 2H), 7.45(d, 1H), 7.25(d, 2H), 7.13 -7.10(m,1H),6.14-6.06(m,2H),5.85-5.82(m,1H),5.58-5.55(m,1H),4.40-4.36(m,1H),4.26-4.22(m, 1H), 4.07-4.00 (m, 3H), 3.89-3.81 (m, 2H), 1.29-1.23 (m, 6H), 1.16-1.11 (t, 3H).

LC-MS m/z: [M+H] ⁺ =598.

Example 55(2S)-2-(((4-(thiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3 Methyl ,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate

4-(thiophen-2-yl)phenol, phosphorus oxychloride, L-alanine methyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxy- 2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was prepared in the same manner as in steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d6) δppm: 11.48(s, 1H), 7.66(d, 2H), 7.58-7.51(m, 2H), 7.46(d, 1H), 7.26(d, 2H), 7.13 (m,1H),6.15-6.00(m,2H),5.87(m,1H),5.56(m,1H),4.41-4.37(m,1H),4.28-4.21(m,1H),4.05(m , 1H), 4.01-3.80 (m, 2H), 3.60 (s, 3H), 1.29-1.17 (m, 6H).

LC-MS m/z: [M+H] ⁺ =584.

Example 56 (2S)-2-(((4-(thiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dioxo-3 ,4-Dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionate neopentyl

4-(thiophen-2-yl)phenol, phosphorus oxychloride, L-alanine neopentyl ester hydrochloride, pentafluorophenol and (2'R)-2'-deoxygenate obtained in Example 26, step 1 -2'-Fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d6) δppm: 11.51(s, 1H), 7.65(d, 2H), 7.58-7.51(m, 2H), 7.45(d, 1H), 7.26(d, 2H), 7.14 -7.11(m,1H),6.17-6.00(m,2H),5.86(d,1H),5.58(d,1H),4.40-4.21(m,2H),4.05-3.98(m,1H),3.93 -3.85(m, 3H), 3.64(m, 1H), 1.31-1.21(m, 6H), 0.86(s, 9H).

LC-MS m/z: [M+H] ⁺ =640.

Example 57 (2S)-2-(((4-(3-methylthiophen-2-yl)phenyloxy)(((2R,3R,4R,5R)-5-(2,4-di Oxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propionic acid neopentyl ester

4-(3-Methylthiophen-2-yl)phenol, phosphorus oxychloride, L-alanine neopentyl ester hydrochloride, pentafluorophenol and (2'R)- 2'-Deoxy-2'-fluoro-2'-methyluridine was used as the raw material, and the target compound was obtained by the same method as steps 2 and 3 of Example 1.

¹ H NMR (500MHz, DMSO-d6) δppm: 11.50(s, 1H), 7.58(d, 1H), 7.45(m, 3H), 7.30(d, 2H), 7.00(d, 1H), 6.17-6.02 (m,2H),5.87(m,1H),5.56(m,1H),4.40-4.21(m,2H),4.05-3.98(m,1H),3.93-3.85(m,3H),3.64(m , 1H), 2.26 (s, 3H), 1.31-1.23 (m, 6H), 0.87 (s, 9H).

LC-MS m/z: [M+H] ⁺ =654.

Pharmacological activity evaluation

Experimental Example 1 Detection of antiviral activity of the compounds of the present invention in the HCV replication system

1. Experimental materials

1.1 Reagents:

Table 1 List of reagents

1.2Huh71b cell line:

The Huh71b cell line, provided by Shanghai WuXi AppTec New Drug Development Co., Ltd., is a Huh7 cell line containing the HCV 1b replicon with a stable luciferase (Luc) reporter. It is constructed by cloning HCV non-structural protein gene, neo (G418 resistance) and luciferase reporter gene into pBR vector through gene recombination technology. Then, the vector carrying the HCV replicon was transfected into huh7 cells, and through G418 resistance screening, the HCV replicon could be stably replicated and related proteins and luciferase were stably expressed in huh7 cells. This cell model is used for in vitro screening of anti-HCV compounds. The anti-HCV activity of the compounds was determined by examining the level of chemiluminescence of the luciferase luminescent substrate. See Lohmann V, et al. 1999. Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line. Science. 285(5424): 110-113.

2. Experimental method:

1) Compound preparation: using the compounds prepared in the above examples of the present invention, each compound was prepared into a stock solution with DMSO, and then diluted to 10 μM with DMEM complete culture medium containing 0.5% DMSO as the compound concentration, 3-fold dilution, a total of 10 Compounds were added to 96-well plates with POD 810 automatic microplate pretreatment system (LabCyte, USA) at different concentrations; at the same time, an ineffective control group (complete medium containing 0.5% DMSO instead of compounds) and 100 % effective control group (only complete culture medium containing 0.5% DMSO was added);

2.1 Cell preparation: Collect log-phase Huh71b cells, resuspend them in DMEM complete culture medium, and inoculate into 96-well plates, 125 μl per well, 8×10 ³ cells/well, placed at 37°C, 5% CO ₂ Cells were cultured in an incubator for 72 hours;

2.2 Cell viability detection: add 30 μl of cell growth fluorescence titration detection reagent to each well, incubate the cells for 1 hour at 37°C, 5% CO ₂ incubator, and detect the fluorescence signal value on a spectrophotometer, and the obtained data is used for compound cytotoxicity calculation;

2.3 Bright-Glo detection: add 100 μl of luciferase luminescent substrate Bright-Glo to each well, and use chemiluminescence detection system EnVision (PerkinElmer, USA) to detect the fluorescence signal value within 5 minutes, and the obtained data is used for compound activity calculation.

2.4 Data processing: Convert the obtained data to percent cell viability (Viability%) using the following formula:

CPD: Fluorescence signal value of compound wells

HPE (Hundred percent effect): 100% effective control fluorescence signal value

ZPE (Zero percent effect): no effect control fluorescence signal value

The raw data was processed as a percent inhibition (Inhibition %) using the following formula:

CPD: Chemiluminescence signal value of compound wells

HPE (Hundred percent effect): 100% effective chemiluminescence signal value

ZPE (Zero percent effect): Invalid effect chemiluminescence signal value

The percent inhibition was imported into GraphPad Prism for further processing to generate corresponding curves and _EC50 values.

The experimental results show that the EC ₅₀ of the compound of the present invention is between about 0.05 μM-1 μM, and has a very good ability to inhibit HCV virus. Some data are shown in Table 2.

Table 2

Example number EC₅₀(μM) Example number EC₅₀(μM) Example 1 0.3266 Example 2 0.1312 Example 3 0.3912 Example 4 1.188

Example 5 0.4984 Example 6 0.4834 Example 9 0.665 Example 10 0.1498 Example 11 0.695 Example 12 0.240 Example 13 0.772 Example 14 0.380 Example 15 0.523 Example 17 0.443 Example 18 0.062 Example 19 0.557 Example 20 0.0783 Example 21 0.173 Example 22 0.104 Example 23 0.265 Example 24 0.055 Example 25 0.204 Example 26 0.019 Example 27 0.169 Example 28 0.142 Example 31 0.103 Example 32 0.174 Example 33 0.130 Example 34 0.196 Example 35 0.051 Example 36 0.146 Example 42 0.065 Example 46 0.055 Example 49 0.049 Example 50 0.063 Example 51 0.018 Example 52 0.021 Example 53 0.097 Example 54 0.020 Example 55 0.032 Example 56 0.028 Example 57 0.030

As announced on Gilead's official website, the current best anti-HCV drug, sofosbuvir (GS7977), has an EC ₅₀ value of 0.102 μM for genotype 1b in the HCV replicon assay, see http://www.gilead for details. com/~/media/Files/pdfs/medicines/liver-disease/sovaldi/sovaldi_pi.pdf. The compounds of the present invention have anti-HCV virus activity equivalent or superior to sofosbuvir.

Experimental Example 2 Detection of antiviral activity of compounds of the present invention on HCV cell infection model (Cell-culture-derived infectious HCV, HCVcc)

1 Experimental material

1.1 Compounds

In this experimental example, the compounds of the present invention prepared in the above examples were used to prepare a 10 mM stock solution with DMSO, and then diluted to 1000 nM with DMEM complete medium containing 0.5% DMSO, and then diluted 3 times successively, with a total of 8 concentrations.

1.2 Cells

Huh 7.5.1 cells were provided by Shanghai WuXi AppTec New Drug Development Co., Ltd.

1.3 Viruses

J399EM (HCV genotype 2a) virus, that is, a full-length HCV mutant strain transfected with EGFP (enhanced green fluorescent protein), has the same infectivity as the JFH-1 wild type. At the same time, by inserting the EGFP coding sequence in the NS5A region, it can Direct observation of NS5A-EGFP fusion protein fluorescence in infected cells, provided by Shanghai WuXi AppTec New Drug Development Co., Ltd.

1.4 Reagents

Reagent name supplier DMEM cell culture medium Invitrogen fetal bovine serum Sigma penicillin-streptomycin Invitrogen Phosphate buffer Hyclone Pancreatin Invitrogen Dimethyl Sulfoxide (DMSO) Sigma Fluorescence assay detection reagent (Renilla luciferase substrate) Promega Cell Viability Assay Reagent (Alamar Blue) Invitrogen

2 Experimental methods

2) Preparation of Huh 7.5.1 cells: collect Huh 7.5.1 cells in log phase, resuspend them in DMEM complete medium, and inoculate them in a 96-well plate (7×10 ³ cells/well) at 37°C , 5% CO ₂ incubator overnight;

3) Drug treatment: The compounds of the present invention were added to the 96-well plate respectively, the compound concentration was 1000nM from beginning to end, and each compound was made into double wells, 3-fold dilution, a total of 8 concentrations, the final concentration of DMSO was 0.5%; Ineffective control group (complete medium containing 0.5% DMSO instead of compound) and 100% effective control group (adding cells without virus infection);

4) Virus infection: J399EM virus supernatant was added to a 96-well plate at a concentration of MOI 0.2 per well. It was then placed in a 37°C, 5% _CO2 incubator for 3 days.

5) Determination of antiviral activity: after culturing, luciferase luminescent substrate was added to each well, and the chemiluminescence detection system Envision was used to detect the Luminescence signal value, and the obtained data was used for the calculation of compound inhibition of HCV activity, and the calculation formula was:

Inhibition%=(RLUs- _RLUc )/( _RLUv - _{RLUc )} ×100

Among them, RLU _V represents the chemiluminescence intensity of the virus control group (complete medium containing 0.5% DMSO instead of the compound), RLU _S represents the chemiluminescence intensity of the corresponding compound treatment group, and RLU _C represents the cell control group (cells without virus infection) chemiluminescence intensity;

6) Cytotoxicity assay: The cell and compound treatment methods were the same as the above steps, but the medium was added to the experimental plate instead of the virus. After culturing in a 5% CO ₂ incubator for 3 days under the same conditions, AlamarBlue, a cell viability detection reagent, was added, and the Fluorescence signal value was detected by a spectrophotometer. The obtained data is used for compound cytotoxicity calculation, and the calculation formula is:

Viability%=RFU _S /RFU _M ×100

Wherein, RFU _M represents the fluorescence intensity of the vehicle control group (complete medium containing 0.5% DMSO instead of compound), and RFU _S represents the fluorescence intensity of the corresponding compound treatment group;

7) Data processing: The inhibition percentage and cell viability percentage were respectively imported into GraphPad Prism software for data processing, and the corresponding curve of the compound and its HCV inhibitory activity (EC ₅₀ ) and cytotoxicity (CC ₅₀ ) values were obtained.

The experimental results show that the EC ₅₀ values of the compounds of the present invention are basically between ₄₀ nM and 150 nM, and the CC ₅₀ values are all greater _than 1000 nM. Some of the compounds of the invention have smaller _EC50 values, eg Example 26 has an _EC50 of 9.3 nM and a _CC50 >1000 nM. It can be seen that the compound of the present invention has excellent antiviral activity, small cytotoxicity and good safety.

In addition, the inventors of the present invention found that the compound of the present invention has a good plasma protein binding rate, is suitable for medicine, and has a very good clinical application prospect. 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 can be made in the present invention without departing from the spirit and scope of the invention. The scope of the right of the present invention is not limited to the detailed description above, but belongs to the claims.

Claims (5)

1. A compound represented by the general formula I:

wherein:

R₁is methyl;

R₂is fluorine;

R₃is OH;

R₄is methyl;

R₅is selected from C_1-6An alkyl group;

Cy₁is phenyl, optionally substituted with one or more groups selected from fluoro and methyl;

Cy₂selected from pyridine ring, pyrazole ring, thiazole ring, pyrrole ring, isoxazole ring, imidazole ring, thiophene ring and oxazole ring, optionally substituted with a halogen selected from halogen, methyl, C_1-6One or more of alkylaminoacyl and cyano substituted;

wherein the compound is not:

2. the compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula Ia:

wherein:

R_a1selected from pyridine ring, pyrazole ring, thiazole ring, pyrrole ring, isoxazole ring, imidazole ring, thiophene ring and oxazole ring, optionally substituted with a halogen selected from halogen, methyl, C_1-6One or more of alkylaminoacyl and cyano substituted;

R_a2selected from fluorine and methyl; or

R_a1、R_a2Together with the carbon atom to which they are attached form a benzopyrazolyl, benzothiazolyl, benzopyrrole or benzisoxazole ring, which is optionally substituted with methyl;

R_a3、R_a4、R_a5independently selected from fluoro and methyl;

R₁is methyl;

R₂is fluorine;

R₃is OH;

R₄is methyl; and

R₅is selected from C_1-6An alkyl group;

wherein the compound is not:

3. a compound or pharmaceutically acceptable salt thereof, wherein the compound is a compound selected from the group consisting of:

4. a pharmaceutical composition comprising a compound of any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. Use of a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 4 in the manufacture of a medicament for the treatment and/or prophylaxis of hepatitis virus infection.