WO2015101183A1

WO2015101183A1 - Uracil nucleotide analogs, preparation methods therefor and uses thereof

Info

Publication number: WO2015101183A1
Application number: PCT/CN2014/094074
Authority: WO
Inventors: 钟慧娟; 岑均达; 马建斌; 谭松良; 高鹏; 喻红平; 徐耀昌; 马景毅; 王听中; 吕爱锋
Original assignee: 江苏豪森药业股份有限公司; 上海翰森生物医药科技有限公司
Priority date: 2014-01-02
Filing date: 2014-12-17
Publication date: 2015-07-09
Also published as: CN104761604A; CN105829334B; TW201605885A; JP2017502975A; CN105829334A

Abstract

The invention relates to uracil nucleotide analogs, preparation methods therefor and uses thereof. Specifically, the invention provides uracil nucleotide analogic compounds having the following formula (I), stereoisomers or pharmaceutically acceptable salts thereof, preparation methods therefor and uses thereof. These compounds are inhibitors of RNA-dependent RNA viral replication and are useful as inhibitors of HCV NS5B polymerase, as inhibitors of HCV replication, and for the treatment of hepatitis C infection in mammals. The compounds have a wide application prospect and are hopeful to be developed into a new generation of antiviral drugs.

Description

Uracil nucleotide analog and preparation method and application thereof

Technical field

The invention belongs to the field of drug synthesis, and in particular relates to a uracil nucleotide analog and a preparation method and application thereof.

Background technique

The viruses of the Flaviviridae family include at least three different genera: pestiviruses, which cause disease in cattle and pigs; flavirus, which is the leading cause of diseases such as dengue and yellow fever. ; and hepaciviruses, the only member of which is HCV. The Flavivirus genus includes more than 68 members and is grouped based on serological kinship. Clinical symptoms vary and include fever, encephalitis, and hemorrhagic fever. Flaviviruses associated with human diseases of global concern include dengue hemorrhagic fever virus (DHF), yellow fever virus, shock syndrome virus, and Japanese encephalitis virus. Since the HCV genome is similar in structure and phenotype to human flavivirus and prion, it is classified as Flaviviridae HCV. Hepatitis C virus is a positive-strand RNA virus that is surrounded by a lipid-containing capsule in the nucleocapsid and has a spike on the capsule. HCV only has three in vitro cell culture systems: Huh7, Huh7.5, and Huh7.5.1. Hepatitis C virus was first discovered in 1974. In 1989, American scientist Michael Houghton and his colleagues used molecular biology methods to find the genetic sequence of the virus and cloned the hepatitis C virus. The disease and its virus are hepatitis C (Hepatitis C) and hepatitis C virus (HCV).

HCV virions are enveloped positive-strand RNA viruses, HCV-RNA consists of approximately 9500-100 bp, 5' and 3' non-coding regions (NCR) are 319-341 bp, and 27-55 bp, respectively, containing several forward and Inverse repeats, possibly related to gene duplication, in the order of 5'-C-E1-E2-p7-NS2-NS3-NS4-NS5-3', encoding a polyprotein of approximately 3014 amino acids in length Precursor, which can be cleaved into 10 viral proteins by host cell and viral autoprotease, including three structural proteins, a 19KD nucleocapsid protein (or core protein, Core) and two glycoproteins. (E1 protein with a molecular weight of 33 kD and E2 protein with a molecular weight of 72 Kd), p7 encodes a membrane-intrinsic protein whose function may be an ion channel. The non-structural protein fractions include NS2, NS3, NS4A, NS5A and NS5B, and the life cycle of non-structural proteins versus viruses is very important. NS2 and NS3 have protease activity and are involved in the cleavage of viral polyprotein precursors. In addition, NS3 protein also has helicase activity, involved in the unwinding of HCV-RNA molecules to assist RNA replication, and the function of NS4 is unclear. NS5A is a phosphoprotein that interacts with a variety of host cell proteins and plays an important role in viral replication. While NS5B has RNA-dependent RNA polymerase activity and is involved in HCV genome replication, NS5B polymerase is considered to be an essential component of the HCV replication complex. Inhibition of HCV NS5B polymerase prevents the formation of double-stranded HCV RNA and thus constitutes an attractive approach to the development of HCV-specific antiviral therapies.

HCV has significant heterogeneity and high variability, and the HCV strains of all known genomic sequences are analyzed to have large differences in nucleotide and amino acid sequences. And the degree of variation in the HCV genome is not consistent, such as 5'-CR is the most conservative, the homology is 92-100%, and the 3'NCR region is highly variable. Among the HCV coding genes, the C region is the most. The conserved, non-structural (NS) region is second, and the capsular membrane protein E2/NS1 has the highest variability called the hypervariable region. It is known that most HCV-I infections are found in European and American countries, while Asian countries are mainly type II, followed by type III. Okomoto reported that Japanese chronic hepatitis C patients and healthy blood donors were mainly type II infections, accounting for 59.3% and 82.4%, respectively, while about 50% of hemophilia patients were type I infections due to the application of imported US factor VIII. Wang reported that 86.2% of patients with chronic hepatitis C in Beijing were type II infections, and type III infections were 13.8%. In Xinjiang, type III infection accounts for 50%, indicating that different types of HCV have certain regional and population distribution characteristics. In addition, different genotype infections cause different clinical processes and interferon treatment responses. For example, type III infection has severe clinical symptoms and causes severe punishment for liver disease: type II (Simmonds 1b) infection is insensitive to interferon therapy, III Type infection (Simononds 2a) is effective with interferon.

Hepatitis C virus (HCV) has severely jeopardized human health, causing chronic liver diseases such as cirrhosis and hepatocellular carcinoma in a large number of infected individuals (estimated to be 2-15% of the world's population). According to the US Centers for Disease Control, there are 4.5 million people infected in the United States alone. According to the World Health Organization, there are more than 200 million infected individuals worldwide, and at least 3 to 4 million people are infected each year. Once infected, approximately 20% of people can clear the virus, but the remaining people may carry HCV for the rest of their lives. 10% to 20% of chronically infected individuals eventually develop liver-destructive sclerosis or cancer. The viral disease is transmitted parenterally through contaminated blood and blood products, contaminated needles; or through sexual transmission; and from the infected mother or carrier mother to their offspring. Current treatments for HCV infection are limited to recombinant interferon alpha alone or in combination with the nucleoside analog ribavirin immunotherapy, which has limited clinical benefit.

The pathogenesis of hepatitis C is still not fully understood. When HCV replicates in hepatocytes, it causes changes in liver cell structure and function or interferes with hepatocyte protein synthesis, which can cause degeneration and necrosis of hepatocytes, indicating that HCV directly damages the liver and causes a certain effect. However, most scholars believe that cellular immunopathological reactions may play an important role. It is found that hepatitis C is similar to hepatitis B, and its tissue infiltrating cells are mainly CD3+. Cytotoxic T cells (TC) specifically attack target cells of HCV infection, which can cause liver. Cell damage. Whether it is acute hepatitis C or chronic hepatitis C, the standard treatment regimen is peginterferon (α-2a or α-2b) in combination with ribavirin. This is also the only effective treatment for hepatitis C. Peginterferon alfa is administered once a week, and the number of administrations is greatly reduced, which is convenient for patients to take medicine. Compared with ordinary interferon three times a week or once every other day, pegylated interferon is also called long-acting interferon. The direct comparison of two long-acting interferons combined with ribavirin showed that the recurrence rate of 12KD peginterferon alfa-2b was significantly lower than that of 40KD peginterferon alfa-2a. The activity of the virus and the distribution of the drug caused by the molecular size are related. It is generally believed that the higher the molecular weight of polyethylene glycol, the lower the antiviral activity, the activity of 12KD peginterferon alfa-2b is significantly higher than that of 40KD long-acting interferon; moreover, 12KD long-acting interferon can Systemic distribution, not only removes the main virus in the liver, but also removes extrahepatic viruses such as lymph nodes, kidneys, spleen, adrenal glands, and salivary glands. Therefore, the recurrence rate after stopping the drug is low. The 40KD macromolecular pegylated interferon is limited to vascular and intrahepatic distribution due to its large molecular size, which is detrimental to viral clearance outside the liver. Not only does it increase the burden on the liver, it is slow to excrete, and because it is not excreted by the kidneys, it is difficult to withdraw the drug when an adverse reaction occurs. It is generally believed that 12KD peginterferon alfa-2b should be used as a priority for the treatment of hepatitis C due to the publication of the head-to-head comparison of the IDEAL test results.

Currently, there are limited treatment options for individuals infected with the hepatitis C virus. The treatment options approved today are the use of recombinant interferon alpha alone or in combination with the nucleoside analog ribavirin. This therapy is limited by its clinical effectiveness and only 50% of the treated patients respond to the therapy. Therefore, there is a need to develop more effective and novel therapies to address the unmet medical needs caused by HCV infection.

Some potential molecular targets that can be identified as drugs for anti-HCV therapeutics, including but not limited to NS2-NS3 autoprotease, N3 protease, N3 helicase and NS5B polymerase. RNA-dependent RNA polymerase is absolutely important for single-stranded RNA genome replication, which has attracted significant interest from pharmaceutical chemists. A nucleoside inhibitor of NS5B polymerase can be used as a non-native substrate that causes chain termination, or as a competitive inhibitor that competes with nucleotides for binding to a polymerase. In order to function as a chain terminator, the nucleoside analog must be taken up by the cell and converted to a triphosphate in vivo to compete for the polymerase nucleotide binding site. This conversion of the triphosphate is typically mediated by cellular kinases that impose additional structural requirements on potential nucleoside polymerase inhibitors. Unfortunately, this limits the direct evaluation of nucleosides as inhibitors of HCV replication to cell-based assays capable of in situ phosphorylation.

In some cases, the biological activity of the nucleoside is impeded by poor substrate properties relative to one or more kinases required to convert the nucleoside to the active triphosphate form. of. The formation of a monophosphate by nucleoside kinase is generally considered to be the rate limiting step in the triphosphorylation process. In order to avoid phosphorylation of the first step in the metabolism of nucleosides to active triphosphate analogs, stable phosphate prodrug preparations have been reported in the literature. The nucleoside phosphoramidate prodrug is a precursor of the active nucleoside triphosphate and should be used to inhibit viral replication in whole cells infected with the virus.

The use of restricted nucleosides as viable therapeutic agents is also their sometimes poor physicochemical and pharmacokinetic properties. These poor properties can limit intestinal absorption of the agent and limit uptake into target tissues or cells. In order to improve their properties, prodrugs of the nucleosides were employed. It has been demonstrated that nucleoside phosphoramidate prodrugs improve the systemic uptake of nucleosides, and further, the phosphoramidate portion of these "pro-nucleotides" is masked by neutral lipophilic groups to obtain a suitable partition coefficient to optimize uptake and The transport into the cells, thereby significantly increasing the intracellular concentration of the nucleoside monophosphate analog relative to the parent nucleoside alone. Enzymatically mediated hydrolysis of the phosphate moiety produces a nucleoside monophosphate that does not require an initial rate of monophosphorylation.

The patents for studying such nucleoside monophosphate analogues in recent years are mainly WO2008121634A2, WO2010075517A2 developed by PHARMASSET, WO2010135520A1 developed by CHIMERIX, WO2012040127A1, WO2012088155A1 developed by ALIOS BIOPHARMA, WO2012142075A1, WO2012142085A1, WO2013009737A1 developed by MERCK SHARP&DOHME CORP. .

Summary of the invention

The inventors discovered a class of uracil nucleotide analogues during the course of the study. These novel compounds are inhibitors of RNA-dependent RNA viral replication and are useful as inhibitors of HCV NS5B polymerase, inhibitors of HCV replication, and It has broad application prospects in the treatment of hepatitis C infection in mammals and is expected to be developed into a new generation of antiviral drugs.

In one aspect, the invention provides a uracil nucleotide analog of the compound of formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

Wherein Z is selected from oxygen or sulfur; Y is selected from hydrogen or acetyl;

R _{1 is} selected from the group consisting of hydrogen, halogen, hydroxy, decyl, cyano, nitro, azide, C _1-8 alkyl, halo-substituted C _1-8 alkyl, C _2-8 alkenyl, C _2-8 Alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _{5 10} aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 C(O)NR ₆ R ₇ , -N(R ₅ )-C(O)R ₅ , -N(R ₅ )-C(O)OR ₅ , -C _0-8 - SR ₅ , -SiR ₅ R ₆ R ₇ , -GeR ₅ R ₆ R ₇ ,

or

The carbon atom adjacent to the benzene ring of R ₁ forms a 5-7 membered carbon ring and a 5-7 membered heterocyclic ring.

Wherein C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, C _5-10 aryl, 5-10 membered heteroaryl, 5-7 membered carbocyclic or 5-7 membered heterocyclic ring optionally further substituted with one or more substituents selected from halo, hydroxy, mercapto, cyano, nitro, azido, C _{1- 8-} alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclic oxy, 3-8 Heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5 -10-membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 - C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N(R _₅₎ -C (O) R ₅ or _{-N (R 5) -C (O} ) oR ₅ is substituted with a substituent;

R _{2 is} selected from the group consisting of hydrogen, halogen, hydroxy, decyl, cyano, nitro, azide, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 ring Alkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _{5 10} arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _{0 -8} -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ Substituted by

Wherein the C _1-8 alkyl group, the C _3-8 cycloalkyl group, the 3-8 membered heterocyclic group, the C _5-10 aryl group or the 5-10 membered heteroaryl group is optionally further selected by one or more From halogen, hydroxy, thiol, cyano, nitro, azide, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3- 8-membered heterocyclic group, 3-8 membered heterocyclic oxy group, 3-8 membered heterocyclic thio group, C _5-10 aryl group, C _5-10 aryloxy group, C _5-10 arylthio group 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ Substituted with a substituent of -C _0-8 -C(O)NR ₆ R ₇ , -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ;

R _{3 is} selected from hydrogen, C _1-8 alkyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, C _5-10 aryl or 5-10 membered heteroaryl, optionally further by one or a plurality selected from the group consisting of halogen, hydroxy, decyl, cyano, nitro, azide, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl , 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic thio group, C _5-10 aryl, C _5-10 aryloxy, C _5-10 aryl Thiothio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR a substituent of ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ Replace

R _{4 is} selected from the group consisting of hydrogen, halogen, hydroxy, decyl, cyano, nitro, azide, C _1-8 alkyl, halo-substituted C _1-8 alkyl, C _3-8 cycloalkyl, C _3-8 Cycloalkylmethyl, halo-substituted C _1-8 alkoxy, halo-substituted C _1-8 alkylthio, 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic ring Thiothio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 N-heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C( O) OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N(R ₅ ) -C(O)R ₅ , -N(R ₅ )-C(O)OR ₅ ;

R ₅ , R ₆ , and R _{7 are} selected from the group consisting of hydrogen, C ₄ -alkyl, C _3-8 cycloalkyl;

m is 0, 1, 2, 3, 4.

r is 0, 1, 2

As used herein, "5-7 membered carbocyclic ring" refers to a percarbocyclic ring containing from 5 to 7 carbon atoms, including cycloalkyl or aryl groups, and "5-7 membered heterocyclic ring" means one or more rings. The atom is selected from heteroatoms of nitrogen, oxygen or S(O)r (wherein r is an integer of 0, 1, 2), and the remaining ring atoms are ring groups of carbon containing 5 to 7 ring atoms.

As a preferred embodiment, when Z is selected from oxygen, its structural formula is a compound of formula (II),

Wherein: Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I).

As a further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, R _{1 is} selected from a C _1-8 alkyl group, a halogen-substituted C _1-8 alkyl group, C _2-8 alkenyl group, C _2-8 alkynyl group, C _3-8 cycloalkyl group, 3-8 membered heterocyclic group, 3-8 membered heterocyclic oxy group, 3-8 membered heterocyclic thio group , C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroaryl Thiothio group, -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N(R ₅ )-C(O)R ₅ , -N(R ₅ )-C (O)OR ₅ , -C _0-8 -SR ₅ , -SiR ₅ R ₆ R ₇ , -GeR ₅ R ₆ R ₇ ,

Wherein C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, C _5-10 aryl or The 5-10 membered heteroaryl group is further optionally further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide, C _1-8 alkyl, C _2-8 alkenyl, C _{2 -8} alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC (O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N(R ₅ )-C(O)R ₅ or -N(R ₅ ) Substituted by a substituent of -C(O)OR ₅ ; Y, R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I).

As a further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, R _{1 is} selected from a C _1-8 alkyl group, a halogen-substituted C _1-8 alkyl group, C _2-8 alkenyl, C _2-8 alkynyl,

Wherein the C _1-8 alkyl group, the halogen-substituted C _1-8 alkyl group, the C _2-8 alkenyl group or the C _2-8 alkynyl group is further one or more selected from the group consisting of C _3-8 cycloalkyl groups , 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic thio group, C _5-10 aryl, C _5-10 aryloxy, C _5-10 aryl Substituted by a substituent of a thiol group, a 5-10 membered heteroaryl group, a 5-10 membered heteroaryloxy group or a 5-10 membered heteroarylthio group; Y, R ₂ , R ₃ , R ₄ , R ₅ And R ₆ , R ₇ , m, r are as defined for the compound of formula (I).

As a still further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

Most preferably, the uracil nucleotide analog, its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

As a further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, R _{1 is} selected from C _3-8 cycloalkyl, 3-8 membered heterocyclic group, 3 -8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl a 5-10 membered heteroaryloxy group, a 5-10 membered heteroarylthio group,

Wherein the C _3-8 cycloalkyl group, the 3-8 membered heterocyclic group, the C _5-10 aryl group or the 5-10 membered heteroaryl group is further further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, Cyano, nitro, azide, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3 -8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl , 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 - C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 - Substituted by a substituent of C(O)NR ₆ R ₇ , -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ;

Y, R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I).

As a still further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a drug thereof The acceptable salt is selected from the group consisting of:

As a further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, R _{1 is} selected from -C _0-8 -SR ₅ , -SiR ₅ R ₆ R ₇ , -GeR ₅ R ₆ R ₇ , Y, R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I).

As a further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof is selected from the group consisting of the following compounds:

As a most preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

As a further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, the carbon atom adjacent to the benzene ring of R ₁ forms a 5-7 membered carbocyclic ring, 5-7 Metacyclic ring,

Wherein the 5-7 membered carbocyclic or 5-7 membered heterocyclic ring is optionally further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide, _C1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic thio , C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered Arylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O) OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N(R ₅ )-C Substituting (O) a substituent of R ₅ or -N(R ₅ )-C(O)OR ₅ ;

Y, R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I).

As a still further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, the carbon atom adjacent to the benzene ring of R ₁ forms a 5-7 membered carbocyclic ring, 5- The 7-membered heterocycle is selected from the following structures:

Wherein the 5-7 membered carbocyclic or 5-7 membered heterocyclic ring is optionally further selected from one or more selected from the group consisting of halogen, hydroxy, decyl, cyano, nitro, azide, _C1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic thio , C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered Arylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O) OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N(R ₅ )-C Substituting (O) a substituent of R ₅ or -N(R ₅ )-C(O)OR ₅ ;

Most preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R ₁ and a carbon atom adjacent to the benzene ring form a 5-7 membered carbocyclic ring, 5-7 members The heterocyclic ring is selected from the following structures:

Preferably, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, Z is selected from the group consisting of sulfur, a compound of the formula (III),

Wherein Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I).

As a further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, R _{4 is} selected from the group consisting of halogen, hydroxy, thiol, C _1-8 alkyl, halogen substituted C _{1 -8} alkyl, C _3-8 cycloalkyl, C _3-8 cycloalkanyl, halogen substituted C _1-8 alkoxy, halogen substituted C _1-8 alkylthio; Y, R ₁ , R ₂ And R ₃ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I).

As a still further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, R _{3 is} selected from the group consisting of hydrogen, C _1-8 alkyl, C _3-8 cycloalkyl a 3-8 membered heterocyclic group, a C _5-10 aryl group or a 5-10 membered heteroaryl group, optionally further selected from one or more selected from the group consisting of halogen, hydroxy, decyl, C _1-8 alkyl, C _{3 - 8} -cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, -C _0-8 -S(O)rR ₅ , -C _0- Substituted by a substituent of ₈ -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ or -C _0-8 -OC(O)R ₅ ; _{4 is} selected from the group consisting of fluorine, methyl, trifluoromethyl, cyclopropyl, cyclopropylmethyl; Y, R ₁ , R ₂ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I) .

As a still further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, R _{3 is} selected from hydrogen, C _1-4 alkyl, cyclopropyl, cyclohexyl or Phenyl, optionally further selected from one or more selected from the group consisting of halogen, hydroxy, decyl, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O Substituting a substituent of R ₅ , -C _0-8 -C(O)OR ₅ or -C _0-8 -OC(O)R ₅ ; R _{4 is} selected from methyl; Y, R ₁ , R ₂ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I).

As a still further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, R _{1 is} selected from the group consisting of hydrogen, C _1-8 alkyl, halogen-substituted C _1-8 alkane a C _2-8 alkenyl group, a C _2-8 alkynyl group, wherein the C _1-8 alkyl group, a halogen substituted C _1-8 alkyl group, a C _2-8 alkenyl group or a C _2-8 group The alkynyl group is further one or more selected from the group consisting of C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _{5 - 10} aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy or 5-10 membered heteroarylthio Substituted by a substituent; Y, R ₂ , R ₅ , R ₆ , R ₇ , m, r are as defined in claim 1.

As a still further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, R _{1 is} selected from a C _3-8 cycloalkyl group, a 3-8 membered heterocyclic group, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered An aryl group, a 5-10 membered heteroaryloxy group, a 5-10 membered heteroarylthio group, wherein said C _3-8 cycloalkyl group, a 3-8 membered heterocyclic group, a C _5-10 aryl group or The 5-10 membered heteroaryl group is further optionally further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide, C _1-8 alkyl, C _2-8 alkenyl, C _{2 -8} alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC (O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N(R ₅ )-C(O)R ₅ or -N(R Substituting ₅ )-C(O)OR ₅ substituent; Y, R ₂ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I).

As a still further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, R _{1 is} selected from -C _0-8 -SR ₅ , -SiR ₅ R ₆ R ₇ , -GeR ₅ R ₆ R ₇ ; Y, R ₂ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I).

As a still further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, the carbon atom adjacent to the benzene ring of R ₁ forms a 5-7 membered carbocyclic ring, 5- 7-membered heterocycle, wherein said 5-7 membered carbocyclic or 5-7 membered heterocyclic ring optionally further substituted with one or more substituents selected from halo, hydroxy, mercapto, cyano, nitro, azido group, C _{1 -8} alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 a heterocyclic heterothio group, a C _5-10 aryl group, a C _5-10 aryloxy group, a C _5-10 arylthio group, a 5-10 membered heteroaryl group, a 5-10 membered heteroaryloxy group, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N( Substituted by a substituent of R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ; Y, R ₂ , R ₅ , R ₆ , R ₇ , m, r are as defined I) Definition of the compound.

Wherein the 5-7 membered carbocyclic or 5-7 membered heterocyclic ring is optionally further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide, _C1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic thio , C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered Arylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O) OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N(R ₅ )-C Substituting (O) a substituent of R ₅ or -N(R ₅ )-C(O)OR ₅ ; Y, R ₂ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I) .

As a still further preferred embodiment, the uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, is selected from the group consisting of:

As a most preferred embodiment, the aforementioned uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the stereoisomer is in the S configuration, has the following structure:

According to another aspect of the present invention, there is provided a process for the preparation of a uracil nucleotide analog of the compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, comprising the steps of:

When Y is selected from the group consisting of acetyl groups, the optional further includes the following reactions:

Optionally, further comprising column chromatography to obtain the stereoisomer thereof, or obtaining the stereoisomer thereof by the following steps:

A further aspect of the invention provides a pharmaceutical composition comprising a therapeutically effective amount of a uracil nucleotide analog of the compound of formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable Carrier.

A further aspect of the invention provides a uracil nucleotide analogue of the compound of formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, for use in the treatment of hepatitis C virus, A Use of drugs for diseases caused by hepatitis B virus, West Nile virus, yellow fever virus, dengue virus, rhinovirus, poliovirus, bovine viral diarrhea virus or Japanese encephalitis virus infection.

Detailed ways

DETAILED DESCRIPTION: Unless otherwise stated, the following terms used in the specification and claims have the following meanings.

"C _1-8 alkyl" means a straight-chain alkyl group having 1 to 8 carbon atoms and a branched alkyl group, the alkyl group means a saturated aliphatic hydrocarbon group, and C _0-8 means no carbon atom or C. _1-8 alkyl, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl- 2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1 , 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl , 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-di Methylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl , 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethyl Hexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl or various branches thereof Isomers, etc.

The alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any available point of attachment, preferably one or more of the following groups, independently selected from the group consisting of halogen, hydroxy, thiol, Cyano, nitro, azide, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3 -8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl , 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 - C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 - Substituted by a substituent of C(O)NR ₆ R ₇ , -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ;

"Cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, and "C _3-8 cycloalkyl" refers to a cycloalkyl group of 3 to 8 carbon atoms, "5-10 membered ring.""Alkyl" means a cycloalkyl group of 5 to 10 carbon atoms, for example:

Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptene Alkenyl, cyclooctyl and the like.

Polycyclic cycloalkyl groups include spiro, fused, and bridged cycloalkyl groups. "Spirocycloalkyl" refers to a polycyclic group that shares a carbon atom (called a spiro atom) between the monocyclic rings. These may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system. Separating a spirocycloalkyl group according to the number of common spiro atoms between the ring and the ring Non-limiting examples of spirocycloalkyl, bisspirocycloalkyl or polyspirocycloalkyl, spirocycloalkyl include:

"Fused cycloalkyl" refers to an all-carbon polycyclic group in which each ring of the system shares an adjacent pair of carbon atoms with other rings in the system, wherein one or more of the rings may contain one or more double bonds, but None of the rings have a fully conjugated π-electron system. Depending on the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused ring alkyl groups, and non-limiting examples of fused cycloalkyl groups include:

"Bridge cycloalkyl" refers to an all-carbon polycyclic group in which two rings share two carbon atoms that are not directly bonded, which may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system . Depending on the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups. Non-limiting examples of bridged cycloalkyl groups include:

The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring wherein the ring to which the parent structure is attached is a cycloalkyl group, non-limiting examples include indanyl, tetrahydrogen Naphthyl, benzocycloheptyl and the like.

The cycloalkyl group may be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide. , C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryl Oxyl, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , Substituted with a substituent of -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ;

"Heterocyclyl" means a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent wherein one or more of the ring atoms are selected from nitrogen, oxygen or S(O)r (where r is an integer 0, 1, 2 a hetero atom, but excluding the ring portion of -OO-, -OS- or -SS-, the remaining ring atoms being carbon. The "5-10 membered heterocyclic group" means a ring group containing 5 to 10 ring atoms, and the "3-8 membered heterocyclic group" means a ring group containing 3 to 8 ring atoms.

Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl and the like.

Polycyclic heterocyclic groups include spiro, fused, and bridged heterocyclic groups. "Spirocyclyl" refers to a polycyclic heterocyclic group in which one atom (called a spiro atom) is shared between the individual rings, wherein one or more of the ring atoms are selected from nitrogen, oxygen or S(O)r (which Where r is a hetero atom of the integers 0, 1, 2), and the remaining ring atoms are carbon. These may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system. The spirocycloalkyl group is classified into a monospiroheterocyclic group, a dispiroheterocyclic group or a polyspiroheterocyclic group depending on the number of common spiro atoms between the ring and the ring. Non-limiting examples of spirocycloalkyl groups include:

"Fused heterocyclyl" refers to a polycyclic heterocyclic group in which each ring of the system shares an adjacent pair of atoms with other rings in the system, and one or more rings may contain one or more double bonds, but none The ring has a fully conjugated pi-electron system in which one or more ring atoms are selected from the group consisting of nitrogen, oxygen or S(O)r (wherein r is an integer of 0, 1, 2) heteroatoms, the remaining ring atoms being carbon. Depending on the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocycloalkyl groups, and non-limiting examples of fused heterocyclic groups include:

"Bridge heterocyclyl" refers to a polycyclic heterocyclic group in which any two rings share two atoms that are not directly bonded, and these may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system, One or more of the ring atoms are selected from the group consisting of nitrogen, oxygen or S(O)r (wherein r is an integer of 0, 1, 2) heteroatoms, and the remaining ring atoms are carbon. Depending on the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups. Non-limiting examples of bridged cycloalkyl groups include:

The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring wherein the ring to which the parent structure is attached is a heterocyclic group, non-limiting examples comprising:

The heterocyclic group may be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups, independently selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide. , C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryl Oxyl, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , Substituted with a substituent of -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ;

"Aryl" means an all-carbon monocyclic or fused polycyclic (ie, a ring that shares a pair of adjacent carbon atoms) groups having a polycyclic ring of a conjugated π-electron system (ie, having a ring adjacent to a carbon atom) a group, "C _5-10 aryl" means an all-carbon aryl group having 5 to 10 carbons, and "5-10 membered aryl group" means an all-carbon aryl group having 5 to 10 carbons, such as phenyl and Naphthyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring wherein the ring to which the parent structure is attached is an aryl ring, non-limiting examples comprising:

The aryl group can be substituted or unsubstituted. When substituted, the substituent is preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido group, C _{1 -8} alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 a heterocyclic heterothio group, a C _5-10 aryl group, a C _5-10 aryloxy group, a C _5-10 arylthio group, a 5-10 membered heteroaryl group, a 5-10 membered heteroaryloxy group, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N( Substituted by a substituent of R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ;

"Heteroaryl" refers to a heteroaromatic system containing from 1 to 4 heteroatoms including nitrogen, oxygen and a hetero atom of S(O)r (wherein r is an integer of 0, 1, 2), 5- A 7-membered heteroaryl group means a heteroaromatic system having 5 to 7 ring atoms, and a 5-10 membered heteroaryl group means a heteroaromatic system having 5 to 10 ring atoms, such as furyl, thienyl, pyridyl, Pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and the like. The heteroaryl ring may be fused to an aryl, heterocyclic or cycloalkyl ring wherein the ring to which the parent structure is attached is a heteroaryl ring, non-limiting examples comprising:

The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide. , C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryl Oxyl, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , Substituted with a substituent of -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ;

"Alkenyl" means an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, and a C _2-8 alkenyl group means a straight or branched olefin containing from 2 to 8 carbons base. For example, vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, and the like.

Alkenyl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido group, C _{1 -8} alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 a heterocyclic heterothio group, a C _5-10 aryl group, a C _5-10 aryloxy group, a C _5-10 arylthio group, a 5-10 membered heteroaryl group, a 5-10 membered heteroaryloxy group, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N( Substituted by a substituent of R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ;

"Alkynyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, and _C2-8 alkynyl refers to a straight or branched alkynyl group containing from 2 to 8 carbons. . For example, ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, and the like.

Alkynyl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido group, C _{1 -8} alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 a heterocyclic heterothio group, a C _5-10 aryl group, a C _5-10 aryloxy group, a C _5-10 arylthio group, a 5-10 membered heteroaryl group, a 5-10 membered heteroaryloxy group, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , -N( Substituted by a substituent of R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ;

"Alkoxy" means -O-(alkyl) wherein alkyl is as defined above. The C _1-8 alkoxy group means an alkyloxy group having 1-8 carbons, and the non-limiting examples include a methoxy group, an ethoxy group, a propoxy group, a butoxy group and the like.

The alkoxy group may be optionally substituted or unsubstituted, and when substituted, the substituent, preferably one or more of the following groups, independently selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide , C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryl Alkoxy, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ ,- C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ Substituting a substituent of -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ;

"Cycloalkoxy" refers to -O-(unsubstituted cycloalkyl) wherein cycloalkyl is as defined above. The C _3-8 cycloalkoxy group means a cycloalkyloxy group having 3-8 carbons, and the non-limiting examples include a cyclopropoxy group, a cyclobutoxy group, a cyclopentyloxy group, a cyclohexyloxy group and the like.

The alkoxy group may be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide. , C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryl Oxyl, 5-10 membered heteroarylthio, -C _0-8 -S(O)rR ₅ , -C _0-8 -OR ₅ , -C _0-8 -C(O)R ₅ , -C _0-8 -C(O)OR ₅ , -C _0-8 -OC(O)R ₅ , -C _0-8 -NR ₆ R ₇ , -C _0-8 -C(O)NR ₆ R ₇ , Substituted with a substituent of -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ;

"Halo-substituted C _1-8 alkyl" means a hydrogen on the alkyl group optionally substituted with 1-8 carbon alkyl groups substituted by fluorine, chlorine, bromine or iodine atoms, such as difluoromethyl, dichloromethyl Base, dibromomethyl, trifluoromethyl, trichloromethyl, tribromomethyl, and the like.

The hydrogen on the "halo-substituted C _1-8 alkoxy" alkyl group is optionally a 1-8 carbon alkoxy group substituted with a fluorine, chlorine, bromine or iodine atom. For example, difluoromethoxy, dichloromethoxy, dibromomethoxy, trifluoromethoxy, trichloromethoxy, tribromomethoxy, and the like.

"Halogen" means fluoro, chloro, bromo or iodo.

"Optional" or "optionally" means that the subsequently described event or environment may, but need not, occur, including where the event or environment occurs or does not occur. For example, "heterocyclic group optionally substituted by an alkyl group" means that an alkyl group may be, but not necessarily, present, and the description includes the case where the heterocyclic group is substituted with an alkyl group and the case where the heterocyclic group is not substituted with an alkyl group. .

"Substituted" refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3, hydrogen atoms, independently of each other, substituted by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art will be able to determine (by experiment or theory) substitutions that may or may not be possible without undue effort. For example, an amino group or a hydroxyl group having a free hydrogen may be unstable when combined with a carbon atom having an unsaturated (e.g., olefinic) bond.

"Pharmaceutical composition" means a mixture comprising one or more of the compounds described herein, or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, as well as other components such as physiological/pharmaceutically acceptable carriers. And excipients. The purpose of the pharmaceutical composition is to promote the administration of the organism, which facilitates the absorption of the active ingredient and thereby exerts biological activity.

The present invention is further described in detail with reference to the accompanying drawings, but by no way of limitation,

The structure of the compound of the present invention is determined by nuclear magnetic resonance (NMR) or/and liquid chromatography-mass spectrometry (LC-MS). The NMR chemical shift (δ) is given in parts per million (ppm). The NMR was measured by a Bruker AVANCE-400 nuclear magnetic apparatus, and the solvent was deuterated dimethyl sulfoxide (DMSO-d ₆ ). The deuterated methanol (CD ₃ OD) and deuterated chloroform (CDCl ₃ ) were labeled as the top four. Silane (TMS).

LC-MS was determined by LC-MS using an Agilent 1200 Infinity Series mass spectrometer. The HPLC was measured using an Agilent 1200 DAD high pressure liquid chromatograph (Sunfire C18 150 x 4.6 mm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18 150 x 4.6 mm column).

The thin layer chromatography silica gel plate uses Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate. The specification for TLC is 0.15mm~0.20mm, and the specification for separation and purification of thin layer chromatography is 0.4mm~0.5mm. Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as a carrier.

The starting materials in the examples of the invention are known and commercially available or can be employed Or synthesized according to methods known in the art.

Unless otherwise stated, all reactions of the present invention were carried out under continuous magnetic stirring under a dry nitrogen or argon atmosphere, and the solvent was a dry solvent.

An argon atmosphere or a nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon having a volume of about 1 L. The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon of about 1 L volume.

The solution in the examples means an aqueous solution unless otherwise specified. The temperature of the reaction is room temperature. The room temperature is an optimum reaction temperature of 20 ° C to 30 ° C.

Monitoring of the progress of the reaction in the examples using a thin layer chromatography (TLC) or liquid chromatography coupled to liquid chromatography (LC-MS) reaction using a solvent system: dichloromethane and methanol system, n-hexane and ethyl acetate system, The volume ratio of the petroleum ether and ethyl acetate systems, acetone, and solvent can be adjusted depending on the polarity of the compound. Column chromatography eluent system includes: A: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: dichloromethane and ethyl acetate system, D: ethyl acetate and methanol, solvent The volume ratio is adjusted depending on the polarity of the compound, and may be adjusted by adding a small amount of ammonia water and acetic acid.

Embodiment 1

First step Preparation of isopropyl ((2-cyclopropylphenoxy)(4-nitrophenoxy)phosphoryl)-L-alanine ester

4-nitrophenyl two phosphorus trichloride (320mg, 1.25mmol) was dissolved in CH _₂ Cl ₂ (2.5mL), cooled to -78 ℃, 2- cyclopropyl-phenol (185mg, 1.38mmol) and TEA (192μL, A solution of 1.38 mmol) in CH ₂ Cl ₂ (2.5 mL) was added dropwise at this temperature, and the reaction was gradually heated to 0 ° C for 30 minutes. The reaction solution was dropwise added to the cooled L-alanine at 0 ° C. isopropyl ester hydrochloride (210mg, 1.25mmol) in CH _₂ Cl ₂ (2.5mL) solution, followed by TEA (366μL, 2.63mmol) was added dropwise to the reaction system was stirred at 0 ℃ 1 hour and concentrated under reduced pressure The reaction mixture was stirred with EtOAc EtOAc. Column chromatography (eluent: PE: EtOAc = 5:1) gave the title compound isopropyl((2-cyclopropylphenoxy)(4-nitrophenoxy)phosphino)-L-propylamine Acid ester (395 mg, 70%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.16-8.23 (m, 2H), 7.33-7.42 (m, 3H), 7.05-7.15 (m, 2H), 6.86-6.93 (m, 1H), 4.95- 5.04(m,1H),3.95-4.18(m,2H),2.02-2.11(m,1H),1.34-1.43(m,3H),1.17-1.30(m,6H),0.83-0.98(m,2H) ), 0.61-0.72 (m, 2H).

The second step is isopropyl ((2-cyclopropylphenoxy) (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidine-1 (2H) Preparation of p-fluoro-4-hydroxy-4-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine

1-((2R,3R,4R,5S)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)- The diketone (58 mg, 0.22 mmol) was dissolved in a mixed solvent of THF (2 mL) and NMP (0.6 mL), and a solution of ^t BuMgCl (1 M, 0.45 mL, 0.45 mmol) was added dropwise to the above solution at room temperature. After stirring for 10 minutes, isopropyl ((2-cyclopropylphenoxy)(4-nitrophenoxy)phosphino)-L-alanine ester (200 mg, 0.45 mmol) was added dropwise to the reaction. A solution of THF (1.5 mL) was stirred at 55 ° C overnight. Then cooled to room temperature, methanol (1 mL) the reaction was quenched, concentrated and column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 30: 1) under reduced pressure to give the title compound isopropyl ((2- cyclopropylmethyl Phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxyl 4-Methyltetrahydrofuran-2-yl)methoxy)phosphino)-L-alanine (15 mg, 12%, epimer ratio: S _P /R _P =4.1:1).

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.80 (s, 1H), 7.44 (d, J = 8.0 Hz, 1H), 7.35 (d, J = 8.0 Hz, 1H), 7.06-7.18 (m, 2H) ), 6.89-6.95 (m, 1H), 6.17 (d, J = 19.6 Hz, 1H), 5.67 (d, J = 8.0 Hz, 1H), 4.95 - 5.07 (m, 1H), 4.43-4.60 (m, 2H), 3.81-4.16 (m, 4H), 2.04-2.15 (m, 1H), 1.18-1.41 (m, 12H), 0.93-1.02 (m, 2H), 0.64-0.77 (m, 2H);

³¹ P NMR (162 MHz, CDCl ₃ ): δ 4.40, 3.50;

MS m/z (ESI): 570.1 [M+H] ⁺ .

Embodiment 2

First step Preparation of isopropyl ((3-cyclopropylphenoxy)(4-nitrophenoxy)phosphoryl)-L-alanine ester

4-nitrophenyl two phosphorous oxychloride (1.950g, 7.62mmol) was dissolved in CH _₂ Cl ₂ (15mL), cooled to -78 ℃, 3- cyclopropyl-phenol (1.124g, 8.38mmol) and TEA (1.17 A solution of mL 2, 8.39 mmol) in CH ₂ Cl ₂ (15 mL) was added dropwise at this temperature for 30 minutes, then gradually warmed to 0 ° C. The reaction solution was added dropwise to the cooled L-alanine at 0 ° C. acid isopropyl ester hydrochloride (1.279g, 7.63mmol) in CH _₂ Cl ₂ (15mL) solution, followed by TEA (2.23mL, 16.0mmol) was added dropwise to the reaction system was stirred at 0 ℃ 1 hour under reduced pressure The reaction mixture was concentrated, EtOAc (EtOAc)EtOAc. Column chromatography (eluent: PE: EtOAc = 4.5:1) gave the title compound isopropyl ((3-cyclopropylphenoxy)(4-nitrophenoxy)phosphino)-L-propylamine Acid ester (2.905 g, 85%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.17-8.26 (m, 2H), 7.33-7.44 (m, 2H), 7.15-7.23 (m, 1H), 6.96-7.04 (m, 1H), 6.86- 6.94 (m, 2H), 4.93-5.05 (m, 1H), 3.92-4.18 (m, 2H), 1.80-1.91 (m, 1H), 1.38 (d, J = 6.8 Hz, 3H), 1.18-1.28 ( m, 6H), 0.93-0.99 (m, 2H), 0.63-0.69 (m, 2H).

The second step is isopropyl ((3-cyclopropylphenoxy) (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidine-1 (2H) Preparation of p-fluoro-4-hydroxy-4-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine

1-((2R,3R,4R,5S)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)- Diketone (272 mg, 1.05 mmol) was dissolved in a mixed solvent of THF (8 mL) and NMP (2.7 mL), and a solution of ^t BuMgCl (1 M, 2.1 mL, 2.10 mmol) was added dropwise to the above solution at room temperature. After stirring for 10 minutes, isopropyl ((3-cyclopropylphenoxy)(4-nitrophenoxy)phosphino)-L-alanine ester (938 mg, 2.09 mmol) was added dropwise to the reaction. A solution of THF (6 mL) was stirred at 55 ° C overnight. Then cooled to room temperature, methanol (3mL) The reaction was quenched, concentrated and column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 30: 1) under reduced pressure to give the title compound isopropyl ((3cyclopropylmethoxy Phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxyl 4-Methyltetrahydrofuran-2-yl)methoxy)phosphino)-L-alanine (20 mg, 3%, epimer ratio: S _P /R _P = 3.8:1).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.74 (s, 1H), 7.46 (d, J = 8.0Hz, 1H), 7.15-7.25 (m, 1H), 6.82-7.07 (m, 3H), 6.09 -6.27 (m, 1H), 5.51-5.75 (m, 1H), 4.94-5.10 (m, 1H), 4.37-4.59 (m, 2H), 3.70-4.16 (m, 4H), 1.79-1.95 (m, 1H), 1.18-1.41 (m, 12H), 0.93-1.10 (m, 2H), 0.63-0.72 (m, 2H);

³¹ P NMR (162 MHz, CDCl ₃ ): δ 4.08, 3.40;

MS m/z (ESI): 570.1 [M+H] ⁺ .

Embodiment 3

First step Preparation of isopropyl ((2-cyclopropyl-6-methylphenoxy)(4-nitrophenoxy)phosphino)-L-alanine ester

4-nitrophenyl two phosphorous oxychloride (1.950g, 7.62mmol) was dissolved in CH _₂ Cl ₂ (15mL), cooled to -78 ℃, 2- cyclopropyl-6-methylphenol (1.129g, 7.62mmol And TEA (1.17mL, 8.39mmol) in CH ₂ Cl ₂ (15mL) solution was added dropwise, reacted at this temperature for 30 minutes, gradually warmed to 0 ° C, the reaction solution was dropped into 0 ° C to cool the L- alanine isopropyl ester hydrochloride (1.279g, 7.63mmol) in CH _₂ Cl ₂ (15mL) solution, followed by TEA (2.23mL, 16.0mmol) was added dropwise to the reaction system was stirred at 0 ℃ The reaction mixture was concentrated under reduced pressure. EtOAc (30 mL). Column chromatography (eluent: PE: EtOAc = 5:1 to 3:1) afforded the title compound isopropyl ((2-cyclopropyl-6-methylphenoxy) (4-nitrophenoxy) Phospho)-L-alanine ester (2.880 g, 82%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.18 (d, J = 9.2Hz, 2H), 7.32 (dd, J = 8.4,7.2Hz, 2H), 7.00 (d, J = 4.8Hz, 2H), 6.8. ), 1.13-1.42 (m, 12H), 0.90-1.06 (m, 2H), 0.59-0.75 (m, 2H).

The second step is isopropyl ((2-cyclopropyl-6-methylphenoxy) ((2R, 3R, 4R, 5R)-5-(2,4-dicarbonyl-3,4-dihydrol Preparation of pyrimidine-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine ester

1-((2R,3R,4R,5S)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)- Diketone (272 mg, 1.05 mmol) was dissolved in a mixed solvent of THF (8 mL) and NMP (2.7 mL), and a solution of ^t BuMgCl (1 M, 2.1 mL, 2.10 mmol) was added dropwise to the above solution at room temperature. After stirring for 10 minutes, isopropyl ((2-cyclopropyl-6-methylphenoxy)(4-nitrophenoxy)phosphino)-L-alaninate was added dropwise to the reaction. (967 mg, 2.09 mmol) in THF (6 mL) EtOAc. Then cooled to room temperature, methanol (3mL) The reaction was quenched, concentrated and column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 30: 1) under reduced pressure to give the title compound isopropyl ((2- cyclopropylmethyl (6-methylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4- Fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine ester (20 mg, 3%, epimer ratio S _P /R _P >10) :1).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.53 (d, J = 8.0 Hz, 1H), 6.98-7.06 (m, 2H), 6.69-6.77 (m, 1H), 6.01-6.08 (m, 1H) ), 5.59 (d, J = 8.0 Hz, 1H), 4.57 (s, 2H), 4.41-4.50 (m, 1H), 4.31-4.40 (m, 1H), 4.03-4.11 (m, 1H), 3.80- 4.02 (m, 2H), 2.38 (s, 3H), 2.24 - 2.35 (m, 1H), 1.26-1.40 (m, 6H), 1.22 (d, J = 6.0 Hz, 6H), 0.95-1.02 (m, 2H), 0.59-0.71 (m, 2H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.81;

MS m/z (ESI): 584.1 [M+H] ⁺ .

Embodiment 4

First step Preparation of isopropyl ((4-cyclopropylphenoxy)(4-nitrophenoxy)phosphoryl)-L-alanine ester

4-nitrophenyl two phosphorous oxychloride (1.260g, 4.92mmol) was dissolved in CH _₂ Cl ₂ (10mL), cooled to -78 deg.] C, 4- cyclopropyl-phenol (726mg, 5.41mmol) and TEA (0.76mL , 5.45 mmol) of CH ₂ Cl ₂ (10 mL) solution was added dropwise, and the reaction was carried out at this temperature for 30 minutes, then gradually warmed to 0 ° C, and the reaction solution was dropwise added to the cooled L-alanine at 0 ° C. isopropyl ester hydrochloride (826mg, 4.93mmol) in CH _₂ Cl ₂ (10mL) solution, followed by TEA (1.44mL, 10.33mmol) was added dropwise to the reaction system was stirred at 0 ℃ 1 hour and concentrated under reduced pressure The reaction mixture was poured with EtOAc (30 mL). Column chromatography (eluent: PE: EtOAc = 5:1) gave the title compound isopropyl((4-cyclopropylphenoxy)(4-nitrophenoxy)phosphino)-L-propylamine Acid ester (1.48 g, 67%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.00-8.28 (m, 2H), 7.32-7.45 (m, 2H), 6.71-7.18 (m, 4H), 4.91-5.02 (m, 1H), 3.93- 4.19 (m, 2H), 1.77-1.92 (m, 1H), 1.39 (d, J = 6.4 Hz, 3H), 1.7-1.26 (m, 6H), 0.90-0.98 (m, 2H), 0.58-0.66 ( m, 2H).

The second step is isopropyl ((4-cyclopropylphenoxy) (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidine-1 (2H) Preparation of p-fluoro-4-hydroxy-4-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine

1-((2R,3R,4R,5S)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)- Diketone (272 mg, 1.05 mmol) was dissolved in a mixed solvent of THF (8 mL) and NMP (2.70 mL), and a solution of ^t BuMgCl (1 M, 2.1 mL, 2.10 mmol) was added dropwise to the above solution at room temperature. After stirring for 10 minutes, isopropyl ((4-cyclopropylphenoxy)(4-nitrophenoxy)phosphino)-L-alanine ester (938 mg, 2.09 mmol) was added dropwise to the reaction. A solution of THF (6 mL) was stirred at 55 ° C overnight. Then cooled to room temperature, methanol (3mL) The reaction was quenched, concentrated and column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 30: 1) under reduced pressure to give the title compound isopropyl ((4-cyclopropyl Phenyloxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxyl 4-Methyltetrahydrofuran-2-yl)methoxy)phosphino)-L-alanine ester (62 mg, 10%, epimer ratio: S _P / R _P = 3.1:1).

¹ H NMR (400 MHz, CDCl ₃ ): δ 9.26-9.45 (m, 1H), 7.25-7.52 (m, 1H), 7.05-7.12 (m, 2H), 6.97-7.04 (m, 2H), 6.16 ( d, J = 18.4 Hz, 1H), 5.54-5.75 (m, 1H), 4.94-5.05 (m, 1H), 4.36-5.57 (m, 2H), 3.70-4.24 (m, 4H), 1.17.91 ( m, 1H), 1.15 - 1.39 (m, 12H), 0.88 - 0.98 (m, 2H), 0.55 - 0.65 (m, 2H);

³¹ P NMR (162 MHz, CDCl ₃ ): δ 4.05, 3.58;

MS m/z (ESI): 570.1 [M+H] ⁺ .

Embodiment 5

First step Preparation of (4-bromo-2-methylphenoxy)(tert-butyl)dimethylsilane

4-bromo-2-methylphenol (2.1 g, 11 mmol) and imidazole (2.2 g, 33 mmol) were dissolved in DMF (10 mL), cooled to 0 ° C, tert-butylchlorodimethylsilane (2.0 g, 14 mmol) It was added to the above solution with stirring. The solution was warmed to room temperature and stirred for 3 h. EtOAc (EtOAc) (EtOAc) The EtOAc layer was washed with EtOAcqqqqHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH Phenoxy)(tert-butyl)dimethylsilane (2.4 g, 73%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ7.05 (m, 1H), 6.94 (m, 1H), 6.43 (d, J = 8.4Hz, 1H), 1.97 (s, 3H), 0.81 (s, 9H ), 0.00(s, 6H).

Second step Preparation of tert-butyl(4-cyclopropyl-2-methylphenoxy)dimethylsilane

Potassium phosphate (6.6 g, 32 mmol) was dissolved in water (10 mL), cyclopropylboronic acid (2.1 g, 24 mmol) and Pd(OAc) ₂ (290 mg, 1.28 mmol) were added to the above solution under stirring, and then continued to be added ( A solution of 4-bromo-2-methylphenoxy)(tert-butyl)dimethylsilane in toluene (50 mL). The suspension was deoxygenated with nitrogen for 45 minutes, and tricyclohexylphosphine (0.9 g, 3.2 mmol) was added to the above solution, and the suspension was reacted at 95 ° C overnight under stirring with nitrogen. The residue was purified by EtOAc (EtOAc) (EtOAc) (EtOAcjjjjjjj : PE) gave the crude title compound t-butyl(4-cyclopropyl-2-methylphenoxy)dimethylsilane (1.1 g, crude yield 50%, 85% purity).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ6.65 (d, J = 2.0Hz, 1H), 6.57 (m, 1H), 6.46 (d, J = 8.0Hz, 1H), 1.98 (s, 3H), 1.61 (m, 1H), 0.81 (s, 9H), 0.68 (m, 2H), 0.41 (m, 2H), 0.00 (s, 6H).

The third step 4-preparation of cyclopropyl-2-methylphenol

To a flask of tert-butyl(4-cyclopropyl-2-methylphenoxy)dimethylsilane (1.1 g, 85% purity, 4 mmol) was added tetrabutylammonium fluoride (1M in THF, 12 mL) , 12mmol). The solution was stirred at room temperature for 2 hours and TLC showed the material disappeared. The solution was diluted with aq. EtOAc (30 mL)EtOAc The EtOAc layer was washed with EtOAc EtOAc EtOAc (HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH 0.64 g, crude yield 80%).

^{1 H NMR (400MHz, DMSO-} d6): δ8.98 (s, 1H), 6.79 (d, J = 1.6Hz, 1H), 6.75 (m, 1H), 6.67 (d, J = 8.0Hz, 1H) , 2.10 (s, 3H), 1.77 (m, 1H), 0.84 (m, 2H), 0.55 (m, 2H).

Fourth step Preparation of isopropyl ((4-cyclopropyl-2-methylphenoxy)(4-nitrophenoxy)phosphoryl)-L-alanine ester

4-Nitrophenylphosphoric dichloride ester (900 mg, 3.6 mmol) was dissolved in CH ₂ Cl ₂ (7.5 mL), cooled to -78 ° C, 4-cyclopropyl-2-methylphenol (600 mg) A solution of TEA (0.39 g, 3.9 mmol) in CH ₂ Cl ₂ (7.5 mL) was added dropwise to the above solution over ten minutes and allowed to warm to room temperature with stirring. The above solution was added dropwise to a solution of isopropyl L-alanine hydrochloride (600 mg, 3.6 mmol) in CH ₂ Cl ₂ (7.5 mL), EtOAc (EtOAc) The reaction system was dropped into the mixture within 5 minutes. The reaction was stirred at 0<0>C for 1 h, EtOAc (EtOAc) (EtOAc)EtOAc. 3) The title compound isopropyl((4-cyclopropyl-2-methylphenoxy)(4-nitrophenoxy)phosphanyl)-L-alanine (550 mg, 33%) was obtained.

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.22 (m, 2H), 7.37 (m, 2H), 7.20 (m, 1H), 6.90 (s, 1H), 6.84 (m, 1H), 5.02 (m , 1H), 4.09 (m, 1H), 3.89 (m, 1H), 2.21 (m, 3H), 1.82 (m, 1H), 1.40 (m, 3H), 1.23 (m, 6H), 0.93 (m, 2H), 0.62 (m, 2H).

Step 5 Isopropyl ((4-cyclopropyl-2-methylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydrol) Preparation of pyrimidine-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (145 mg, 0.56 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.45 mL). ^t BuMgCl (1.0 M in THF, 1.1 mL, 1.1 mmol) was added dropwise to the above solution over 5 minutes while stirring, and the reaction was stirred at room temperature for 10 minutes, then isopropyl ((4-cyclopropyl-2-methyl) A solution of (4-nitrophenoxy)phosphino)-L-alanine ester (550 mg, 1.15 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated to dryness and purified by column chromatography (eluent: CH ₂ Cl ₂ : MeOH = 50:1 to 10:1) to give the title compound isopropyl ((4-cyclopropyl-2-methylphenoxy) (((2R,3R,4R,5R)-5-(2,4-Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyl Tetrahydrofuran-2-yl)methoxy)phosphino)-L-alanine ester (29 mg, 9%, epimer ratio: S _P /R _P = 9.3:1).

^{_{1 H NMR (400MHz, CD 3}} OD): δ7.57 (d, J = 8.0Hz, 1H), 7.21 (m, 1H), 6.97 (s, 1H), 6.88 (m, 1H), 6.13 (d, J=19.6 Hz, 1H), 5.59-5.64 (m, 1H), 4.93-5.01 (m, 1H), 4.49-4.54 (m, 1H), 4.36-4.41 (m, 1H), 4.09-4.13 (m, 1H), 3.89-3.97 (m, 2H), 2.30 (s, 3H), 1.83-1.89 (m, 1H), 1.31-1.40 (m, 7H), 1.23-1.26 (m, 6H), 0.92-0.96 ( m, 2H), 0.62-0.66 (m, 2H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 4.09, 3.99;

MS m/z (ESI): 584.2 [M+H] ⁺ .

Embodiment 6

First step Preparation of 3-(4-(benzyloxy)phenyl)m-butyl ring

(4-(Benzyloxy)phenyl)boronic acid (684 mg, 3.00 mmol), NiI ₂ (28.0 mg, 90 mmol), trans-2-aminocyclohexanol hydrochloride (11.0 mg, 90.0 mmol) dissolved in ⁱ To a solution of <RTI ID=0.0></RTI></RTI><RTIgt;</RTI><RTIgt;</RTI><RTIgt;</RTI><RTIgt;</RTI><RTIgt; The reaction was carried out at 80 ° C for 30 minutes under microwave. The reaction was carried out in parallel in three batches, and the reaction liquids of the three reactions were cooled, combined, diluted with EtOH, filtered over Celite, and the filtrate was concentrated and purified by column chromatography (eluent: PE: EA = 50:1) -(4-(Benzyloxy)phenyl)m-butyl (400 mg, 56%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ7.32 (m, 7H), 6.91 (d, J = 8.8Hz, 2H), 4.97 (m, 4H), 4.68 (t, J = 6.4Hz, 2H), 4.08 (m, 1H).

Step 2 Preparation of 4-(oxabutan-3-yl)phenol

3-(4-(Benzyloxy)phenyl)-propanol ring (400 mg, 1.67 mmol), Pd/C (10 wt%, 50 mg) was mixed with EtOH (20 mL) and reacted under hydrogen atmosphere for 3 hours. Filtration on a short pad of silica gel eluted EtOAcqqqqq

^{_{1 H NMR (400MHz, CDCl 3}} ): δ7.29 (d, J = 8.4Hz, 2H), 6.83 (d, J = 8.4Hz, 2H), 5.05 (m, 2H), 4.74 (m, 2H), 4.19 (m, 1H).

Third step Preparation of isopropyl ((4-nitrophenoxy)(4-(oxabutyl-3-yl)phenoxy)phosphino)-L-alanine ester

4-nitrophenyl two phosphorus trichloride (427mg, 1.67mmol) was dissolved in CH _₂ Cl ₂ (6mL), cooled to -78 deg.] C, 4- (dioxetane-3-yl) phenol (250mg, 1.67mmol) A solution of TEA (233 μL, 1.67 mmol) in CH ₂ Cl ₂ (3 mL) was added dropwise and allowed to react at room temperature for 40 min. Then it was cooled to -78 ° C, and a solution of L-alanine isopropyl ester hydrochloride (280 mg, 1.67 mmol) in CH ₂ Cl ₂ (2.5 mL) and TEA (466 μL, 3.34 mmol) of CH ₂ cl ₂ (2mL) was slowly warmed to room temperature and stirred overnight. The solution was concentrated under reduced pressure. EtOAc (EtOAc)EtOAc. Column chromatography (eluent: PE: EtOAc = 2:1) gave the title compound isopropyl((4-nitrophenoxy)(4-(indobutyl-3-yl)phenoxy)phosphino )-L-Alanine ester (540 mg, 70%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.11 (d, J = 8.4Hz, 2H), 7.30 (m, 4H), 7.16 (m, 2H), 4.92 (m, 3H), 4.62 (t, J = 6.4 Hz, 2H), 4.47 (m, 1H), 4.11 (m, 1H), 1.30 (m, 3H), 1.30 (m, 6H);

MS m/z (ESI): 465.1 [M+H] ⁺ .

The fourth step is isopropyl (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3) Of -hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(oxabutyl-3-yl)phenoxy)phosphino)-L-alanine

1-((2R,3R,4R,5S)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)- The diketone (152 mg, 0.584 mmol) was dissolved in a mixed solvent of THF (4 mL) and NMP (1 mL), and a solution of ^t BuMgCl (1M, 1.17 mL, 1.17 mmol) was added dropwise to the above solution. After stirring at room temperature for 20 minutes, isopropyl ((4-nitrophenoxy)(4-(oxabutyl-3-yl)phenoxy)phosphino)-L-alanine was added dropwise to the reaction. A solution of the ester (542 mg, 1.17 mmol) in THF (2 mL) Then cooled to room temperature, methanol (2mL) The reaction was quenched, concentrated and column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 20: 1) under reduced pressure to give the title compound isopropyl ((((2R, 3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl )methoxy)(4-(oxabutan-3-yl)phenoxy)phosphino)-L-alanine ester (35 mg, 10%, a pair of epimers ratio S _P /R _P = 2.5:1).

^{_{1 H NMR (400MHz, CD 3}} OD): δ7.51 (m, 1H), 7.33 (m, 2H), 7.16 (m, 2H), 6.03 (m, 1H), 5.46-5.56 (m, 1H), 4.97 (m, 2H), 4.88 (m, 1H), 4.61 (m, 2H), 4.45 (m, 1H), 4.30 (m, 1H), 4.18 (m, 1H), 4.01 (m, 1H), 3.83 (m, 2H), 1.25 (m, 6H), 1.11 (m, 6H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.93, 3.89;

MS m/z (ESI): 586.2 [M+H] ⁺ .

Example 7

First step Preparation of 4-(cyclopropylmethyl)phenol

To a solution of 4-hydroxy-phenylcyclopropyl ketone (4.2 g, 26 mmol) in THF (15 mL), EtOAc (EtOAc) Then, boron trifluoride diethyl ether (0.32 mL, 2.6 mmol) was added and stirring was continued for 1 hour. After the TLC reaction was completed, the reaction mixture was poured into ice water, the organic phase was separated, and the organic phase was dried over anhydrous magnesium sulfate and evaporated. (Cyclopropylmethyl)phenol (3.7 g, 96%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ7.13 (d, J = 8.4Hz, 2H), 6.80 (d, J = 8.4Hz, 2H), 5.35 (brs, 1H), 2.48 (d, J = 6.8 Hz, 2H), 0.94 (m, 1H), 0.51 (m, 2H), 0.18 (m, 2H).

Second step Preparation of isopropyl ((4-(cyclopropylmethyl)phenoxy)(4-nitrophenoxy)phosphoryl)-L-alanine ester

Dry ice in acetone (-78 ° C), to a solution of 4-nitrophenylphosphoric acid dichloride (1.50 g, 5.86 mmol) in CH ₂ Cl ₂ (30 mL) ester hydrochloride (982mg, 5.86mmol) and TEA (1.63mL, 11.7mmol) in CH _₂ Cl ₂ (15mL) was added dropwise, slowly warmed to room temperature, and stirring continued for 1 hour. It was cooled to -78 ° C with dry ice-acetone, and then a solution of 4-(cyclopropylmethyl)phenol (868 mg, 5.86 mmol) in CH ₂ Cl ₂ (5 mL) and TEA (0.82 mL, 5.86 mmol). Rin slowly to room temperature and stir overnight. The reaction mixture was washed with EtOAc EtOAc (EtOAc m. (Cyclopropylmethyl)phenoxy)(4-nitrophenoxy)phosphino)-L-alanine ester (180 mg, 6.7%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.17 (m, 2H), 7.32 (m, 2H), 7.17 (m, 2H), 7.07 (m, 2H), 4.96 (m, 1H), 4.00 (m , 1H), 2.44 (m, 2H), 1.36 (m, 3H), 1.17 (m, 6H), 0.85 (m, 1H), 0.47 (m, 2H), 0.12 (m, 2H);

MS m/z (ESI): 463.0 [M+H] ⁺ .

Step 3 Isopropyl ((4-(cyclopropylmethyl)phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidine) Preparation of -1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)alaninate

1-((2R,3R,4R,5S)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)- Diketone (51 mg, 0.20 mmol) was dissolved in a mixed solvent of THF (2 mL) and NMP (0.5 mL), and a solution of ^t BuMgCl (1M, 0.39 mL, 0.39 mmol) was added dropwise to the above solution at room temperature. After stirring for 20 minutes, isopropyl ((4-(cyclopropylmethyl)phenoxy)(4-nitrophenoxy)phosphino)-L-alanine ester (180 mg, 0.39) was added dropwise to the reaction. A solution of mmol in THF (1 mL) was stirred at 55 ° C overnight. Then, it was cooled to room temperature, and the reaction was quenched by the addition of methanol (1 mL), and the organic solvent was concentrated to remove the residue, and the residue was diluted with a mixture of CHCl ₃ and ⁱ PrOH (v: v = 3:1, 20 mL) with saturated brine washed several times, dried over anhydrous sodium sulfate, filtered and concentrated by column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 40: 1), to give the title compound as a colorless foam isopropyl ((4- (cyclopropylmethyl Methyl)phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro- 3-Hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)alaninate (54 mg, 47%, epimer ratio: S _P /R _P = 3.9:1).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.42 (d, J = 8.0 Hz, 1H), 7.07 (d, J = 8.4 Hz, 2H), 6.97 (m, 2H), 5.94 (d, J = 19.2 Hz, 1H), 5.41 (m, 1H), 4.78 (m, 1H), 3.92 (m, 1H), 3.74 (m, 2H), 2.33 (d, J = 6.8 Hz, 2H), 1.18 (m, 3H), 1.03 (m, 3H), 0.75 (m, 1H), 0.32 (m, 2H), 0.01 (m, 2H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.99, 3.90;

MS m/z (ESI): 584.2 [M+H] ⁺ .

Example eight

First step Preparation of isopropyl ((4-nitrophenoxy)(4-(phenylethynyl)phenoxy)phosphino)-L-alanine ester

To a solution of 4-nitrophenylphosphoric acid dichloride (924 mg, 3.61 mmol) in CH ₂ Cl ₂ (10 mL), dry-ice-acetone (-78 ° C) yl) phenol (700mg, 3.61mmol), triethylamine (0.833mL, 3.61mmol) in CH _₂ Cl ₂ (7mL) was added dropwise, warmed to rt and stirring was continued for 40 minutes, re-cooling to -78 deg.] C, successively A solution of L-alanine isopropyl ester hydrochloride (605 mg, 3.61 mmol) in CH ₂ Cl ₂ (5 mL) and a solution of TEA (1.67 mL, 7.22 mmol) in CH ₂ Cl ₂ (5 mL) When finished, slowly warm to room temperature and stir overnight. The reaction mixture was concentrated. EtOAc was evaporated. :1) The title compound isopropyl((4-nitrophenoxy)(4-(phenylethynyl)phenoxy)phosphino)-L-alanine (1.0 g, 55%) was obtained.

¹ H NMR (400 Hz, CDCl ₃ ): δ 8.17 (dd, J = 8.8, 2.0 Hz, 2H), 7.45 (m, 4H), 7.31 (m, 5H), 7.16 (t, J = 7.6 Hz, 2H) ), 4.95 (m, 1H), 4.01 (m, 1H), 3.86 (m, 1H), 1.33 (d, J = 7.2 Hz, 3H), 1.17 (m, 6H);

MS m/z (ESI): 509.0 [M+H] ⁺ .

The second step is isopropyl (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3) Of -hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(phenylethynyl)phenoxy)phosphino)-L-alanine

1-((2R,3R,4R,5S)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)- Diketone (26.0 mg, 0.100 mmol) was dissolved in a mixed solvent of THF (2 mL) and NMP (0.5 mL), and a solution of ^t BuMgCl (1.0 M in THF, 0.2 mL, 0.2 mmol) was slowly added dropwise to the above solution. Stir at room temperature for 20 minutes, and slowly add isopropyl ((4-nitrophenoxy)(4-(phenylethynyl)phenoxy)phosphino)-L-alaninate to the reaction. (102 mg, 0.200 mmol) in THF (1 mL)EtOAc. The next day cooling to room temperature, added MeOH (1mL) the reaction was quenched, and then washed to remove most of the organic solvent was concentrated, the residue was washed with water, filtered, the filter cake was dissolved in CH _₂ Cl _2, dried over anhydrous sodium sulfate, filtered and concentrated prepared thin purification layer _{_{^{(eluent: CH 2 Cl 2: i PrOH}}} = 17: 1), to give the title compound as a colorless foam isopropyl ((((2R, 3R, 4R, 5R) -5- (2,4- Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(phenylethynyl) Phenoxy)phosphoryl)-L-alanine ester (14 mg, 22%, epimer ratio: S _P /R _P = 3.8:1).

¹ H NMR (400 Hz, CD ₃ OD): δ 7.53 (d, J = 8.0 Hz, 1H), 7.41 (m, 4H), 7.27 (m, 3H), 7.20 (d, J = 7.2 Hz, 2H) , 6.01 (m, 1H), 5.56-5.80 (m, 1H), 4.88 (m, 1H), 4.42 (m, 1H), 4.30 (m, 1H), 4.03 (m, 1H), 3.83 (m, 2H) ), 1.26 (m, 6H), 1.06 (m, 6H);

MS m/z (ESI): 630.1 [M+H] ⁺ .

Example nine

First step Preparation of 4-(cyclopropylethynyl)phenol

4-iodophenol (5.00 g, 22.7 mmol), DIPEA (18.0 mL, 109 mmol) was dissolved in DMF (80 mL), and the air was replaced with nitrogen three times, then ethynylcyclopropane (2.50 mL, 29.5 mmol), tetrakis(triphenylphosphine)palladium (1.00 g, 0.866 mmol) and CuI (900 mg, 109 mmol). The reaction was slowly warmed to room temperature and stirred under nitrogen overnight. The reaction mixture was filtered through celite, and then evaporated, evaporated, evaporated, evaporated, evaporated. The title compound: 4-(cyclopropylethynyl)phenol (3.50 g, 97%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ7.28 (dd, J = 7.2,2.0Hz, 2H), 6.75 (dd, J = 7.2,2.0Hz, 2H), 5.21 (s, 1H), 1.45 (m , 1H), 0.78-0.89 (m, 4H).

Second step Preparation of isopropyl ((4-(cyclopropylethynyl)phenoxy)(4-nitrophenoxy)phosphino)-L-alanine ester

4-nitrophenyl two phosphorus trichloride (2.50g, 9.77mmol) was dissolved in CH _₂ Cl ₂ (20mL), cooled to -78 deg.] C, 4- (cyclopropyl ethynyl) phenol (1.55g, 9.77mmol) A solution of TEA (1.36 mL, 9.77 mmol) in CH ₂ Cl ₂ (8 mL) was added dropwise and the mixture was reacted at room temperature for 40 min. Then it was cooled to -78 ° C, and a solution of L-alanine isopropyl ester hydrochloride (1.64 g, 9.77 mmol) in CH ₂ Cl ₂ (5 mL) and TEA (2.72 mL, 19.5 mmol) _A solution of ₂ Cl ₂ (7 mL) was slowly warmed to room temperature and stirred overnight. The solution was concentrated under reduced pressure. EtOAc (EtOAc)EtOAc. Column chromatography (eluent: PE: EtOAc = 5:1) gave the title compound isopropyl((4-(cyclopropylethynyl)phenoxy)(4-nitrophenoxy)phosphino)- L-Alanine ester (3.30 g, 72%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.24 (dd, J = 8.8,2.0Hz, 2H), 7.36 (m, 4H), 7.14 (m, 2H), 5.00 (m, 1H), 4.12 (m , 1H), 1.42 (m, 1H), 1.38 (m, 3H), 1.24 (m, 6H), 0.78-0.89 (m, 4H);

MS m/z (ESI): 473.1 [M+H] ⁺ .

Step 3 Isopropyl ((4-(cyclopropylethynyl)phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidine) Preparation of -1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine

1-((2R,3R,4R,5S)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)- The diketone (52 mg, 0.20 mmol) was dissolved in a mixed solvent of THF (2 mL) and NMP (0.5 mL), and a solution of ^t BuMgCl (1M, 0.40 mL, 0.40 mmol) was added dropwise to the above solution. Stir at room temperature for 20 minutes, and dropwise add isopropyl ((4-(cyclopropylethynyl)phenoxy)(4-nitrophenoxy)phosphino)-L-alanine to the reaction. A solution of the ester (189 mg, 0.40 mmol) in THF (1 mL) Then cooled to room temperature, methanol (1 mL) the reaction was quenched, concentrated and column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 30: 1) under reduced pressure to give the title compound isopropyl ((4- (cyclo Propylethynyl)phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro 3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphino)-L-alanine ester (45 mg, 38%, epimer ratio: S _P /R _P = 3.4: 1).

^{_{1 H NMR (400MHz, CD 3}} OD): δ7.50 (m, 1H), 7.24 (d, J = 8.8Hz, 2H), 7.08 (m, 2H), 6.02 (m, 1H), 5.57 (m, 1H), 4.86 (m, 1H), 4.42 (m, 1H), 4.30 (m, 1H), 4.01 (m, 1H), 3.80 (m, 2H), 1.34 (m, 1H), 1.28 (m, 6H) ), 1.11 (m, 6H), 0.77 (m, 2H), 0.62 (m, 2H);

MS m/z (ESI): 594.3 [M+H] ⁺ .

Example ten

First step Preparation of 4-(propylthio)phenol

Potassium carbonate (28 g, 200 mmol), p-hydroxythiophenol (2.5 g, 20 mmol) and 1-bromopropane (3.1 g, 25 mmol) were added to DMF (40 mL) at room temperature, and the suspension was stirred at room temperature for three days. The disappearance of the starting material was detected by LC-MS. The reaction mixture was poured into water (200 mL), EtOAc (EtOAc m. The title compound 4-(propylthio)phenol (0.9 g, 27%) was obtained.

^{1 H NMR (400MHz, DMSO-} d 6): δ9.52 (s, 1H), 7.21 (d, J = 8.8Hz, 2H), 6.73 (d, J = 8.8Hz, 2H), 2.76 (t, J = 7.2 Hz, 2H), 1.49 (m, 2H), 0.93 (t, J = 7.2 Hz, 1H).

Second step Preparation of isopropyl ((4-nitrophenoxy)(4-(propylthio)phenoxy)phosphino)-L-alanine ester

4-Nitrophenylphosphoric dichloride ester (1.3 g, 5 mmol) was dissolved in CH ₂ Cl ₂ (10 mL) in EtOAc. EtOAc. A solution of 5.6 mmol) and TEA (0.56 g, 5.6 mmol) in CH ₂ Cl ₂ (10 mL) was added dropwise to the above solution over ten minutes and then allowed to warm to room temperature under stirring. The above solution was added dropwise to a solution of isopropyl L-alanine hydrochloride (840 mg, 5 mmol) in CH ₂ Cl ₂ (10 mL) cooled at 0 ° C, then TEA (1.05 g, 10.5 mmol) in 5 min Drip into the reaction system. The reaction was stirred at 0<0>C for 1 h, EtOAc (EtOAc) (EtOAc)EtOAc. 3) The title compound isopropyl((4-nitrophenoxy)(4-(propylthio)phenoxy)phosphino)-L-alanine (1.5 g, 60%) was obtained.

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.16 (m, 2H), 7.33 (m, 2H), 7.24 (m, 2H), 7.08 (m, 2H), 4.94 (m, 1H), 4.02 (m , 1H), 3.86 (m, 1H), 2.78 (m, 2H), 1.58 (m, 2H), 1.33 (m, 3H), 1.16 (m, 6H), 0.94 (t, J = 7.2 Hz, 3H) .

The third step is isopropyl (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3) Of -hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(propylthio)phenoxy)phosphino)-L-alanine

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.3 mL). ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes while stirring, and the reaction was stirred at room temperature for 10 minutes, then isopropyl ((4-nitrophenoxy) ( A solution of 4-(propylthio)phenoxy)phosphino)-L-alanine ester (484 mg, 1 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction was concentrated and taken half column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 50: 1 ~ 10: 1) to give the title compound isopropyl ((((2R, 3R, 4R, 5R) -5- (2,4-Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-( Propylthio)phenoxy)phosphino)-L-alanine ester (66 mg, 22%, epimer ratio: S _P /R _P = 3.5:1).

^{_{1 H NMR (400MHz, CD 3}} OD): δ7.63 (d, J = 8.0Hz, 1H), 7.36 (m, 2H), 7.22 (m, 2H), 6.12 (m, 1H), 5.65-5.72 ( m,1H), 4.54 (m,1H), 4.40-4.42 (m,1H),4.12-4.14 (m,1H),3.91-3.95 (m,2H),2.87-2.92 (m,2H),1.61- 1.66 (m, 2H), 1.31-1.40 (m, 7H), 1.23-1.26 (m, 6H), 1.02 (m, 3H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.97, 3.91;

MS m/z (ESI): 604.2 [M+H] ⁺ .

Embodiment 11

First step Preparation of 4-(tert-butylthio)phenol

To the tert-butyl chloride (25 g) was added p-hydroxythiophenol (3.1 g, 25 mmol) at room temperature. Aluminum trichloride (4 g, 30 mmol) was added in portions, and the reaction was stirred at room temperature for one hour, and the material disappeared by LC-MS. The reaction mixture was poured into EtOAc (EtOAc) (EtOAc)EtOAc. = 4:1) The title compound 4-(tert-butylthio)phenol (3.0 g, 65%) was obtained.

^{1 H NMR (400MHz, DMSO-} d 6): δ9.76 (s, 1H), 7.28 (d, J = 8.8Hz, 2H), 6.77 (d, J = 8.8Hz, 2H), 1.19 (s, 9H ).

Second step Preparation of isopropyl ((4-nitrophenoxy)(4-(tert-butylthio)phenoxy)phosphino)-L-alanine ester

4-Nitrophenylphosphoric dichloride ester (3.8 g, 15 mmol) was dissolved in CH ₂ Cl ₂ (30 mL) in EtOAc. A solution of 3.0 g, 16 mmol) and TEA (1.6 g, 16 mmol) in CH ₂ Cl ₂ (30 mL) was added dropwise to the above solution over ten minutes and allowed to warm to room temperature with stirring. The solution was added dropwise (30mL) cooled at 0 ℃ L- alanine isopropyl ester hydrochloride (2.5g, 15mmol) in CH _₂ Cl ₂ solution, and then TEA (3.2g, 32mmol) in 5 minutes Drip into the reaction system. The reaction was stirred at 0<0>C for 1 h, EtOAc (EtOAc) (EtOAc)EtOAc. 4) The title compound isopropyl((4-nitrophenoxy)(4-(tert-butylthio)phenoxy)phosphino)-L-alanine (3.0 g, 40%) was obtained.

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.23 (m, 2H), 7.50 (m, 2H), 7.40 (m, 2H), 7.19 (m, 2H), 5.01 (m, 1H), 4.07 (m , 1H), 3.96 (m, 1H), 1.40 (m, 3H), 1.22-1.28 (m, 15H).

The third step is isopropyl (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3) Of -hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(tert-butylthio)phenoxy)phosphino)-L-alanine

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.3 mL). ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes while stirring, and the reaction was stirred at room temperature for 10 minutes, then isopropyl ((4-nitrophenoxy) ( A solution of 4-(tert-butylthio)phenoxy)phosphino)-L-alanine ester (500 mg, 1 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated by column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 50: 1 ~ 10: 1) to give the title compound isopropyl ((((2R, 3R, 4R, 5R) -5- (2 ,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(tert-butyl) Thio)phenoxy)phosphino)-L-alanine ester (126 mg, 41%, epimer ratio: S _P /R _P = 2.2:1).

^{_{1 H NMR (400MHz, CD 3}} OD): δ7.66 (m, 1H), 7.53 (m, 2H), 7.28 (m, 2H), 6.14 (m, 1H), 5.67-5.73 (m, 1H), 4.96 (m, 1H), 4.57 (m, 1H), 4.39-4.44 (m, 1H), 4.13-4.15 (m, 1H), 3.90-4.00 (m, 2H), 1.30-1.43 (m, 7H), 1.20-1.27 (m, 14H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.79, 3.74;

MS m/z (ESI): 618.2 [M+H] ⁺ .

Example twelve

First step Preparation of ethyl (4-methoxybenzyl) sulfane

4-Methoxybenzyl mercaptan (5.0 g, 32.5 mmol) was dissolved in DMF (100 mL). EtOAc (EtOAc, m. The reaction was diluted with EtOAc (EtOAc) (EtOAc)EtOAc. The organic phase was separated, dried over anhydrous sodium sulfate, filtered and evaporated tolulujjjjjjjjjjjjjjj g, 67%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ7.26 (d, J = 8.8Hz, 2H), 6.87 (d, J = 8.8Hz, 2H), 3.82 (s, 3H), 3.71 (s, 2H), 2.45 (q, J = 7.2 Hz, 2H), 1.25 (t, J = 7.2 Hz, 3H).

Step 2 Preparation of 4-((ethylthio)methyl)phenol

Ethyl (4-methoxybenzyl) sulfane (3.9 g, 22 mmol) was placed in a 250 mL flask, and a solution of BBr ₃ (1M, 55 mL, 55 mmol) in CH ₂ Cl ₂ (55 mL) was added dropwise to the reaction. After stirring at room temperature for one hour, the disappearance of the starting material was confirmed by LC-MS. And the reaction was diluted with CH _₂ Cl ₂ (50mL) was added saturated sodium bicarbonate solution, the organic phase was washed with saturated brine (100 mL) and dried over anhydrous sodium sulfate, filtered and concentrated by column chromatography (eluent: PE: EtOAc = 9:1 to 6:4) The title compound ethyl (4-methoxybenzyl)sulfane (2.5 g, 67%) was obtained.

^{1 H NMR (400MHz, DMSO-} d 6): δ9.31 (s, 1H), 7.09 (d, J = 8.4Hz, 2H), 6.88 (d, J = 8.4Hz, 2H), 3.82 (s, 2H ), 2.37 (q, J = 7.2 Hz, 2H), 1.15 (t, J = 7.2 Hz, 3H).

Third step Preparation of isopropyl ((4-nitrophenoxy)(4-((ethylthio)methyl)phenoxy)phosphino)-L-alanine ester

4-Nitrophenylphosphoric dichloride ester (3.5 g, 13.5 mmol) was dissolved in CH ₂ Cl ₂ (27 mL) in EtOAc. A solution of phenol (2.5 g, 15 mmol) and TEA (1.5 g, 15 mmol) in CH ₂ Cl ₂ (27 mL) was added dropwise to the above solution over ten minutes and allowed to warm to room temperature with stirring. The above solution was added dropwise to a solution of isopropyl L-alanine hydrochloride (2.3 g, 13.5 mmol) in CH ₂ Cl ₂ (27 mL), then EtOAc (3 g, The reaction system was added dropwise over a minute, and the reaction was slowly warmed to room temperature and stirred overnight. After concentrating the solution, EtOAc (EtOAc (EtOAc)EtOAc. (4-Nitrophenoxy)(4-((ethylthio)methyl)phenoxy)phosphino)-L-alanine ester (1.7 g, 25%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.23 (m, 2H), 7.41 (m, 2H), 7.29 (m, 2H), 7.16 (m, 2H), 5.01 (m, 1H), 4.11 (m , 1H), 3.97 (m, 1H), 3.68 (m, 2H), 2.41 (m, 2H), 1.40 (m, 3H), 1.22 (m, 9H).

The fourth step is isopropyl (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3) Of -hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-((ethylthio)methyl)phenoxy)phosphino)-L-alanine

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (4 mL) and NMP (1.3 mL). ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes while stirring, and the reaction was stirred at room temperature for 10 minutes, then isopropyl ((4-nitrophenoxy) ( A solution of 4-((ethylthio)methyl)phenoxy)phosphino)-L-alanine ester (480 mg, 1 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated by column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 50: 1 ~ 10: 1) to give the title compound isopropyl ((((2R, 3R, 4R, 5R) -5- (2 ,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-((B Thio)methyl)phenoxy)phosphoryl)-L-alanine ester (150 mg, 50%, epimer ratio: S _P /R _P = 2.2:1).

^{_{1 H NMR (400MHz, CD 3}} OD): δ7.63 (d, J = 8.4Hz, 1H), 7.35 (d, J = 8.4Hz, 1H), 7.22 (m, 2H), 6.16 (m, 1H) , 5.64-5.71 (m, 1H), 4.95-5.02 (m, 1H), 4.52-4.57 (m, 1H), 4.37-4.42 (m, 1H), 4.12 (m, 1H), 3.89-3.98 (m, 2H), 3.73 (m, 2H), 2.39-2.46 (m, 2H), 1.31-1.40 (m, 7H), 1.20-1.27 (m, 9H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.93, 3.83;

MS m/z (ESI): 604.2 [M+H] ⁺ .

Example thirteen

First step Preparation of isopropyl ((4-nitrophenoxy)(4-(trimethylsilyl)phenoxy)phosphino)-L-alanine ester

4-Nitrophenylphosphoric dichloride ester (1.02 g, 4 mmol) dissolved in CH ₂ Cl ₂ (8 mL) in EtOAc. A solution of (750 mg, 4.5 mmol) and TEA (0.45 g, 4.5 mmol) in CH ₂ Cl ₂ (8 mL) was added dropwise to the above solution over ten minutes and then allowed to warm to room temperature with stirring. The above solution was added dropwise to a solution of isopropyl L-alanine hydrochloride (690 mg, 4 mmol) in CH ₂ Cl ₂ (8 mL), EtOAc (EtOAc) Into the reaction system. The reaction was stirred at 0<0>C for 1 h, EtOAc (EtOAc) (EtOAc) (EtOAc) The title compound isopropyl((4-nitrophenoxy)(4-(trimethylsilyl)phenoxy)phosphino)-L-alanine (1.2 g, 60%) was obtained.

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.13 (m, 2H), 7.39 (m, 2H), 7.31 (m, 2H), 7.11 (m, 2H), 4.90 (m, 1H), 4.01 (m , 1H), 3.79 (m, 1H), 1.30 (m, 3H), 1.13 (m, 6H), 0.15 (m, 9H).

The second step is isopropyl (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3) Of -hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(trimethylsilyl)phenoxy)phosphino)-L-alanine

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.3 mL). ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes while stirring, and the reaction was stirred at room temperature for 10 minutes, then isopropyl ((4-nitrophenoxy) ( A solution of 4-(trimethylsilyl)phenoxy)phosphino)-L-alanine ester (480 mg, 1.0 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction was concentrated and taken half column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 50: 1 ~ 10: 1) to give the title compound isopropyl ((((2R, 3R, 4R, 5R) -5- (2,4-Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-( Trimethylsilyl)phenoxy)phosphoryl)-L-alanine ester (38 mg, 13%, epimer ratio: S _P /R _P >10:1).

^{_{1 H NMR (400MHz, CD 3}} OD): δ7.49 (d, J = 8.0Hz, 1H), 7.43 (m, 2H), 7.15 (m, 2H), 6.03 (d, J = 20.0Hz, 1H) , 5.48 (m, 1H), 4.83-4.89 (m, 1H), 4.41-4.46 (m, 1H), 4.25-4.31 (m, 1H), 4.00-4.03 (m, 1H), 3.78-3.86 (m, 2H), 1.19-1.28 (m, 7H), 1.10.14.14 (m, 6H), 0.14 (s, 9H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.77;

MS m/z (ESI): 602.2 [M+H] ⁺ .

Embodiment 14

First step Preparation of isopropyl ((2,3-dihydro-1H-indol-5-yl)oxy)(4-nitrophenoxy)phosphino)-L-alanine ester

4-Nitrophenylphosphoric dichloride ester (2.0 g, 8.0 mmol) was dissolved in CH ₂ Cl ₂ (16 mL), cooled to -78 ° C, 2,3-dihydro-1H-indole-5-ol ( A solution of 1.2 g, 9.0 mmol) and TEA (0.9 g, 9 mmol) in CH ₂ Cl ₂ (16 mL) was added dropwise to the above solution over ten minutes and allowed to warm to room temperature with stirring. The above solution was added dropwise to a solution of isopropyl L-alanine hydrochloride (1.35 g, 8 mmol) in CH ₂ Cl ₂ (16 mL), EtOAc (EtOAc) Drip into the reaction system. The reaction was stirred at 0<0>C for 1 h, EtOAc (EtOAc) (EtOAc) (EtOAc) The title compound isopropyl(((2,3-dihydro-1H-indol-5-yl)oxy)(4-nitrophenoxy)phosphino)-L-alanine ester (1.9 g, 50%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.15 (m, 2H), 7.33 (m, 2H), 7.07 (m, 1H), 7.01 (m, 1H), 6.89 (m, 1H), 4.94 (m , 1H), 4.02 (m, 1H), 3.87 (m, 1H), 2.80 (m, 4H), 2.00 (m, 2H), 1.33 (m, 3H), 1.16 (m, 6H).

The second step is isopropyl (((2,3-dihydro-1H-indol-5-yl)oxy) ((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3) Of 4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (260 mg, 1.0 mmol) was dissolved in a mixed solution of THF (8 mL) and anhydrous NMP (2.6 mL). ^t BuMgCl (1.0 M in THF, 2.0 mL, 2.0 mmol) was added dropwise to the above solution over 5 minutes while stirring, and the reaction was stirred at room temperature for 10 minutes, then isopropyl (((2,3-dihydro-1H) A solution of (茚-5-yl)oxo)(4-nitrophenoxy)phosphino)-L-alanine ester (900 mg, 2 mmol) in dry THF (6 mL) The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated to dryness and purified by column chromatography (eluent: CH ₂ Cl ₂ : MeOH = 50:1 to 10:1) to give the title compound isopropyl (((2,3-dihydro-1H-indole-5) -yl)oxo)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3- Hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine ester (40 mg, 7%, epimer ratio: S _P /R _P = 3.3:1).

^{_{1 H NMR (400MHz, CD 3}} OD): δ7.61 (d, J = 8.0Hz, 1H), 7.18 (d, J = 8.0Hz, 1H), 7.12 (s, 1H), 7.01 (d, J = 8.0 Hz, 1H), 6.14 (d, J = 18.8 Hz, 1H), 5.57-5.67 (m, 1H), 4.95-5.02 (m, 1H), 4.52-4.56 (m, 1H), 4.35-4.40 (m , 1H), 4.12 (d, J = 3.0 Hz, 1H), 3.89-3.98 (m, 2H), 2.86-2.92 (m, 4H), 2.06-2.14 (m, 2H), 1.31-1.39 (m, 7H) ), 1.23-1.27 (m, 6H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.96, 3.90;

MS m/z (ESI): 570.2 [M+H] ⁺ .

Example fifteen

First step Preparation of isopropyl ((4-nitrophenoxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphino)-L-alanine ester

4-Nitrophenylphosphoric dichloride ester (600 mg, 2.4 mmol) was dissolved in CH ₂ Cl ₂ (5 mL) in EtOAc. EtOAc. A solution of 2-phenol (400 mg, 2.6 mmol) and TEA (0.26 g, 2.6 mmol) in CH ₂ Cl ₂ (5 mL) was added dropwise to the above solution over ten minutes and then allowed to warm to room temperature under stirring. The above solution was added dropwise to a solution of isopropyl L-alanine hydrochloride (400 mg, 2.4 mmol) in CH ₂ Cl ₂ (5 mL), then EtOAc (5 mg) Drop into the reaction system. The reaction was stirred at 0<0>C for 1 h. EtOAc (EtOAc) (EtOAc)EtOAc. The title compound isopropyl((4-nitrophenoxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphino)-L-alanine ester (480 mg, 44%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.16 (m, 2H), 7.33 (m, 2H), 6.94 (m, 1H), 6.87 (m, 2H), 4.94 (m, 1H), 4.02 (m , 1H), 3.79 (m, 1H), 2.65 (m, 4H), 1.69 (m, 4H), 1.33 (m, 3H), 1.16 (m, 6H).

The second step is isopropyl (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3) -Hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(Preparation of (5,6,7,8-tetrahydronaphthalen-2-yl)oxo)phosphoryl)-L-alanine

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (4 mL) and NMP (1.3 mL). ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes while stirring, and the reaction was stirred at room temperature for 10 minutes, then isopropyl ((4-nitrophenoxy) ( A solution of (5,6,7,8-tetrahydronaphthalen-2-yl)oxophosphino)-L-alanine ester (480 mg, 1 mmol) in dry THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated to dryness and purified by column chromatography (eluent: CH ₂ Cl ₂ : MeOH = 50:1 to 10:1) to give the title compound isopropyl (((2,3-dihydro-1H-indole-5) -yl)oxo)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3- Hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphino)-L-alanine ester (25 mg, 9%, epimer ratio: S _P /R _P = 4.0:1).

^{_{1 H NMR (400MHz, CD 3}} OD): δ7.59 (d, J = 8.4Hz, 1H), 7.04 (m, 1H), 6.96 (m, 2H), 6.14 (d, J = 18.8Hz, 1H) , 5.55-5.65 (m, 1H), 4.95-5.02 (m, 1H), 4.52-4.60 (m, 1H), 4.35-4.40 (m, 1H), 4.11-4.13 (m, 1H), 3.89-3.98 ( m, 2H), 2.74 (m, 4H), 1.77-1.81 (m, 4H), 1.31-1.39 (m, 7H), 1.23-1.28 (m, 6H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.91, 3.86;

MS m/z (ESI): 584.2 [M+H] ⁺ .

Example sixteen

First step isopropyl ((2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxo)(4-nitrophenoxy)phosphanyl)-L - Preparation of alanine ester

4-Nitrophenylphosphoric dichloride ester (600 mg, 2.4 mmol) was dissolved in CH ₂ Cl ₂ (5 mL) in EtOAc. [1,4] Dioxin-6-ol (400 mg, 2.6 mmol) and TEA (0.26 g, 2.6 mmol) in CH ₂ Cl ₂ (5 mL) were added dropwise to the above solution over ten minutes, then stirred under stirring Rise to room temperature. The above solution was added dropwise to a solution of isopropyl L-alanine hydrochloride (400 mg, 2.4 mmol) in CH ₂ Cl ₂ (5 mL), then EtOAc (5 mg) Drop into the reaction system. The reaction was stirred at 0<0>C for 1 h. EtOAc (EtOAc) (EtOAc)EtOAc. The title compound isopropyl ((2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxo)(4-nitrophenoxy)phosphanyl)-L was obtained. -Alanine ester (580 mg, 53%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.25 (m, 2H), 7.41 (m, 2H), 6.80 (m, 2H), 6.74 (m, 1H), 5.03 (m, 1H), 4.24 (m , 4H), 4.11 (m, 1H), 3.89 (m, 1H), 1.42 (m, 3H), 1.25 (m, 6H).

The second step is isopropyl ((2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxo)((2R,3R,4R,5R)-5- (2,4-Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)- L-propyl Preparation of lysine ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (150 mg, 0.6 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.5 mL). ^t BuMgCl (1.0 M in THF, 1.2 mL, 1.2 mmol) was added dropwise to the above solution over 5 minutes while stirring, and the reaction was stirred at room temperature for 10 minutes, then isopropyl (((2,3-dihydrobenzo). [b][1,4] Dioxin-6-yl)oxo)(4-nitrophenoxy)phosphino)-L-alanine ester (580 mg, 1.2 mmol) in THF (3 mL) Instill in five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction solution was concentrated in half and then subjected to column chromatography (eluent: CH ₂ Cl ₂ : MeOH = 50:1 to 10:1) to give 40 mg of product containing a small amount of NMP. Preparation of thin chromatography (EtOAc) was continued to afford the title compound isopropyl (((2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxy) ((2R, 3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl Methoxy)phosphoryl)-L-alanine ester (20 mg, 6%, epimer ratio is S _P /R _P =1.9:1).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.58-7.61 (m, 1H), 6.75-6.81 (m, 2H), 6.69-6.72 (m, 2H), 6.12-6.17 (m, 1H), 5.61 -5.69 (m, 1H), 4.94-5.00 (m, 1H), 4.48-4.53 (m, 1H), 4.32-4.37 (m, 1H), 4.19-4.24 (m, 4H), 4.08-4.11 (m, 1H), 3.86-3.94 (m, 2H), 1.29-1.38 (m, 7H), 1.22-1.26 (m, 6H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 4.16, 4.08;

MS m/z (ESI): 588.1 [M+H] ⁺ .

Example seventeen

First step Preparation of S-isopropyl (2S)-2-((4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanesulfate

Phenyl diphosphide dichloride (1.0 g, 4.7 mmol) was dissolved in CH ₂ Cl ₂ (10 mL) in EtOAc. EtOAc (EtOAc) A solution of g, 5.4 mmol) in CH ₂ Cl ₂ (10 mL) was added dropwise to the above solution over ten minutes and then allowed to warm to room temperature under stirring. The above solution was added dropwise to a solution of S-isopropyl(S)-2-aminopropanesulfate hydrochloride (1.2 g, 4.7 mmol) in CH ₂ Cl ₂ (10 mL). g, 10 mmol) was added dropwise to the reaction system over 5 minutes. The reaction was stirred at 0<0>C for 1 h, EtOAc (EtOAc)EtOAcEtOAcEtOAc 1) The title compound S-isopropyl(2S)-2-((4-nitrophenoxy)(phenoxy)phosphanyl)amino)propylsulfate (1.03 g, 50%) was obtained.

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.16 (m, 2H), 7.33 (m, 2H), 7.28 (m, 2H), 7.16 (m, 3H), 4.09 (m, 1H), 3.86 (m , 1H), 3.53 (m, 1H), 1.33 (m, 3H), 1.21 (m, 6H).

The second step S-isopropyl (2S)-2-(((((2(), 2R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1 (2H)) Of 4-(4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propanesulfate

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.3 mL). ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes while stirring. After stirring at room temperature for 10 minutes, S-isopropyl (2S)-2-((( A solution of 4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanesulfate (424 mg, 1.0 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction was concentrated and taken half column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 50: 1 ~ 10: 1) to give the title compound S- isopropyl (2S) -2 - ((( ((2R, 3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl (Methoxy)(phenoxy)phosphoryl)amino)propanesulfate (66 mg, 24%, epimer ratio: S _P /R _P = 2.2:1).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.62 - 7.66 (m, 1H), 7.38 - 7.42 (m, 2H), 7.21 - 7.31 (m, 3H), 6.15 (d, J = 18.8 Hz, 1H) ), 5.64-5.71 (m, 1H), 4.55-4.59 (m, 1H), 4.40-4.45 (m, 1H), 4.12-4.15 (m, 1H), 3.94-4.02 (m, 2H), 3.49-3.58 (m, 1H), 1.26-1.40 (m, 12H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.66, 3.55;

MS m/z (ESI): 546.1 [M+H] ⁺ .

Example 18

First step S-methyl(S)-2-((tert-butoxycarbonyl)amino)propyl sulfate

In a sealed tube, N-tert-butoxycarbonyl-L-alanine (5.7 g, 30 mmol) and TEA (3.0 g, 30 mmol) were dissolved in THF (100 mL), cooled to -20 ° C, isobutyl chloroformate ( 4.0g, 30mmol) was injected into the above solution in. The solution was stirred at -20 ° C for 1 hour, sodium methanethiolate (1.7 g, 24 mmol) was added to the above solution, and the mixture was stirred at room temperature under a closed tube and stirred overnight. After the reaction mixture was concentrated, the title compound was obtained (jjjjjjjjjjjjj 3.4g, 67%).

^{1 H NMR (400MHz, DMSO-} d 6): δ4.97 (s, 1H), 4.40 (q, J = 7.2Hz, 1H), 2.29 (s, 3H), 1.46 (s, 9H), 1.37 (d , 3H).

Second step Preparation of S-methyl(S)-2-aminopropionate hydrochloride

Add HCl (4N in dioxane, 20 mL) to a flask containing S-methyl(S)-2-((tert-butoxycarbonyl)amino)propylsulfate (3.4 g, 16 mmol) and stir at room temperature for three hours. , LC-MS detected the disappearance of raw materials. The solvent was concentrated to give the title compound md.

^{1 H NMR (400MHz, DMSO-} d 6): δ8.59 (s, 3H), 4.29 (q, J = 7.2Hz, 1H), 2.37 (s, 3H), 1.45 (d, J = 7.2Hz, 3H ).

Third step Preparation of S-methyl(2S)-2-((4-nitrophenoxy)(phenoxy)phosphoryl)amino)propyl sulfate

Phenylphosphoric acid phenyl ester (3.15 g, 15 mmol) was dissolved in CH ₂ Cl ₂ (30 mL) in EtOAc. EtOAc (EtOAc: EtOAc. A solution of g, 16 mmol) in CH ₂ Cl ₂ (30 mL) was added dropwise to the above solution over ten minutes, and then the mixture was allowed to warm to room temperature under stirring. The above solution was added dropwise to a solution of S-methyl(S)-2-aminopropanesulfate hydrochloride (2.3 g, 15 mmol) in CH ₂ Cl ₂ (30 mL). g, 33 mmol) was added dropwise to the reaction system over 5 minutes. The reaction was stirred at 0<0>C for 1 h, EtOAc (EtOAc) (EtOAc)EtOAc. 1) The title compound S-methyl(2S)-2-((4-nitrophenoxy)(phenoxy)phosphanyl)amino)propylsulfate (2.7 g, 45%) was obtained.

The fourth step S-methyl (2S)-2-((((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-) Of 4-(4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propanesulfate

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.3 mL). ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes while stirring. After stirring at room temperature for 10 minutes, S-methyl(2S)-2-((4) A solution of -nitrophenoxy)(phenoxy)phosphoryl)amino)propanesulfate (400 mg, 1 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated by column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 50: 1 ~ 10: 1) to give the title compound isopropyl ((((2R, 3R, 4R, 5R) -5- (2 ,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(tert-butyl) Thio)phenoxy)phosphino)-L-alanine ester (130 mg, 50%, epimer ratio: S _P /R _P = 2.2:1).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.60-7.65 (m, 1H), 7.38-7.42 (m, 2H), 7.21-7.31 (m, 3H), 6.15 (m, 1H), 5.62-5.69 (m, 1H), 4.55-4.59 (m, 1H), 4.40-4.45 (m, 1H), 4.12-4.15 (m, 1H), 3.94-4.06 (m, 2H), 2.25-2.29 (m, 3H) , 1.30-1.40 (m, 6H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.83, 3.54;

MS m/z (ESI): 518.0 [M+H] ⁺ .

Example 19

First step Preparation of S-(tert-butyl)(S)-2-((tert-butoxycarbonyl)amino)propanesulfate

N-tert-Butoxycarbonyl-L-alanine (9.0 g, 48 mmol) and DCC (10.0 g, 48 mmol) were dissolved in CH ₂ Cl ₂ (60 mL) and stirred at 35 ° C for 15 min. Alcohol (3.6 g, 40 mmol) and HOBt (650 mg, 5 mmol) were added to the reaction mixture, and the mixture was stirred at 35 ° C overnight. Suspension was filtered, the filter cake was washed with CH _₂ Cl ₂ (60mL). After concentration under reduced pressure, EtOAc (EtOAc:EtOAc:EtOAc:EtOAc Amino)propyl sulfate (2.2 g, 17%).

¹ H NMR (400 MHz, CDCl ₃ ): δ 5.05 (d, J = 6.8 Hz, 1H), 4.31 (m, 1H), 1.49 (s, 9H), 1.45 (s, 9H), 1.35 (d, J) =7.2Hz, 3H).

Second step Preparation of S-tert-butyl(S)-2-aminopropionate hydrochloride

S-(tert-butyl)(S)-2-((tert-butoxycarbonyl)amino)propanesulfate (2.2 g, 8.5 mmol) was dissolved in 4N aqueous solution of dioxane (20 mL). Stir at 30 ° C for two hours. The disappearance of the starting material was observed by LC-MS, and the solvent was evaporated to give the title compound (yield: s.

^{1 H NMR (400MHz, DMSO-} d 6): δ8.59 (s, 3H), 4.14 (m, 1H), 1.48 (s, 9H), 1.43 (d, J = 7.2Hz, 3H).

Third step Preparation of S-tert-butyl(2S)-2-((4-nitrophenoxy)(phenoxy)phosphoryl)amino)propane sulfate

Phenylphosphoric acid dichloride (1.7 g, 8.0 mmol) was dissolved in CH ₂ Cl ₂ (15 mL) in EtOAc. EtOAc (EtOAc) A solution of 0.9 g, 9.0 mmol of CH ₂ Cl ₂ (15 mL) was added dropwise to the above solution over ten minutes, and then the mixture was allowed to warm to room temperature under stirring. The above solution was added dropwise to a solution of S-tert-butyl(S)-2-aminopropanesulfate hydrochloride (1.7 g, 8.5 mmol) in CH ₂ Cl ₂ (15 mL). g, 18 mmol) was added dropwise to the reaction system over 5 minutes. The reaction was stirred at 0<0>C for 1 h, EtOAc (EtOAc) (EtOAc)EtOAc. 4) The title compound S-tert-butyl(2S)-2-((4-nitrophenoxy)(phenoxy)phosphanyl)amino)propylsulfate (1.9 g, 53%) was obtained.

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.22 (m, 2H), 7.40 (m, 4H), 7.22 (m, 3H), 4.11 (m, 1H), 3.93 (m, 1H), 1.42 (m , 9H), 1.37 (m, 3H).

The fourth step S-tert-butyl (2S)-2-(((((2(), 2R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1 (2H)) Of 4-(4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propanesulfate

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.3 mL). After stirring, ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes. After stirring at room temperature for 10 minutes, S-tert-butyl(2S)-2-((( A solution of 4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanesulfate (440 mg, 1.0 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated by column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 50: 1 ~ 10: 1) to give the title compound S- tert-butyl (2S) -2 - ((( ((2R, 3R, 4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) (oxy)(phenoxy)phosphoryl)amino)propanesulfate (134 mg, 49%, epimer ratio: S _P /R _P = 2.2:1).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.64 - 7.66 (d, J = 8.4 Hz, 1H), 7.38 - 7.42 (m, 2H), 7.21 - 7.30 (m, 3H), 6.17 (m, 1H) ), 5.64-5.71 (m, 1H), 4.54-4.59 (m, 1H), 4.38-4.45 (m, 1H), 4.12-4.15 (m, 1H), 3.88-3.96 (m, 2H), 1.28-1.50 (m, 15H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.49;

MS m/z (ESI): 560.0 [M+H] ⁺ .

Example twenty

First step Preparation of S-cyclohexyl(2S)-2-((4-nitrophenoxy)(phenoxy)phosphino)amino)propanesulfate

Phenylphosphoric acid dichloride (1.7 g, 8.0 mmol) was dissolved in CH ₂ Cl ₂ (15 mL) in EtOAc. EtOAc (EtOAc) A solution of 0.9 g, 9.0 mmol of CH ₂ Cl ₂ (15 mL) was added dropwise to the above solution over ten minutes, and then the mixture was allowed to warm to room temperature under stirring. The above solution was added dropwise to a solution of S-cyclohexyl(S)-2-aminopropanesulfate hydrochloride (1.8 g, 8.0 mmol) in CH ₂ Cl ₂ (15 mL). , 18 mmol) was dropped into the reaction system within 5 minutes. The reaction was stirred at 0<0>C for 1 h, EtOAc (EtOAc) (EtOAc)EtOAc. 4) The title compound S-cyclohexyl(2S)-2-((4-nitrophenoxy)(phenoxy)phosphanyl)amino)propylsulfate (2.5 g, 70%) was obtained.

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.22 (m, 2H), 7.40 (m, 4H), 7.22 (m, 3H), 4.18 (m, 1H), 3.88 (m, 1H), 3.47 (m , 1H), 1.84 (m, 2H), 1.67 (m, 2H), 1.57 (m, 1H), 1.24-1.45 (m, 8H).

The second step S-cyclohexyl (2S)-2-((((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-) Of 4-(4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propanesulfate

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.3 mL). ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes while stirring. After stirring at room temperature for 10 minutes, S-cyclohexyl (2S)-2-((4) A solution of -nitrophenoxy)(phenoxy)phosphoryl)amino)propanesulfate (424 mg, 1.0 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated by column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 50: 1 ~ 10: 1) to give the title compound S- cyclohexyl (2S) -2 - ((( ((2R, 3R, 4R ,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy (Phenoxy)phosphoryl)amino)propanesulfate (122 mg, 42%, epimer ratio: S _P /R _P = 2.2:1).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.62 - 7.66 (m, 1H), 7.38 - 7.42 (m, 2H), 7.21 - 7.31 (m, 3H), 6.15 (d, J = 18.8 Hz, 1H) ), 5.65-5.71 (m, 1H), 4.55-4.59 (m, 1H), 4.40-4.45 (m, 1H), 4.12-4.15 (m, 1H), 3.94-4.02 (m, 2H), 3.40-3.43 (m, 1H), 1.87 (m, 2H), 1.71 (m, 2H), 1.48 (m, 1H), 1.29-1.48 (m, 11H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.64, 3.51;

MS m/z (ESI): 586.2 [M+H] ⁺ .

Embodiment 21

First step Preparation of S-isopropyl (2S)-2-((4-nitrophenoxy)(4-cyclopropylphenoxy)phosphoryl)amino)propanesulfate

4-Nitrophenylphosphonium dichloride (1.75 g, 6.75 mmol) was dissolved in CH ₂ Cl ₂ (15 mL), cooled to -78 ° C, 4-cyclopropyl phenol (1.0 g, 7.5 mmol) and TEA (0.75 g, 7.5 mmol) of CH ₂ Cl ₂ (15 mL) was added dropwise at this temperature, and the reaction was gradually heated to 0 ° C for 30 minutes. The reaction solution was added dropwise to a cooled S-isopropyl group at 0 ° C. A solution of (S)-2-aminopropanesulfate hydrochloride (1.3 g, 6.75 mmol) in CH ₂ Cl ₂ (15 mL), then EtOAc (1,5 g, 15 mmol) was added dropwise to the reaction system and stirred at 0 ° C The reaction mixture was concentrated under reduced pressure. EtOAc (EtOAc) Column chromatography (eluent: PE: EtOAc = 9:1 to 1.5:1) toield of the title compound S-isopropyl(2S)-2-(((4-nitrophenoxy)) Phenoxyl)phosphoryl)amino)propanesulfate (2.1 g, 70%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.20-8.23 (m, 2H), 7.36-7.41 (m, 2H), 7.09-7.13 (m, 2H), 7.01-7.04 (m, 2H), 4.13- 4.17 (m, 1H), 3.89-3.93 (m, 1H), 3.57-3.61 (m, 1H), 1.86-1.88 (m, 1H), 1.28-1.40 (m, 3H), 1.24-1.28 (m, 6H) ), 0.92-0.98 (m, 2H), 0.62-0.66 (m, 2H).

The second step S-isopropyl (2S)-2-(((((2(), 2R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1 (2H)) Of 4-(4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-cyclopropylphenoxy)phosphoryl)amino)propanesulfate

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.3 mL). ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes while stirring. After stirring at room temperature for 10 minutes, S-isopropyl (2S)-2- ((4) A solution of (4-nitrophenoxy)(4-cyclopropylphenoxy)phosphino)amino)propanesulfate (464 mg, 1.0 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated by column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 50: 1 ~ 10: 1) to give the title compound S- isopropyl (2S) -2 - ((( ((2R, 3R, 4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) (oxy)(4-cyclopropylphenoxy)phosphino)amino)propanesulfate (60 mg, 20%, epimer ratio: S _P /R _P = 4.7:1).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.59-7.62 (m, 1H), 7.16-7.18 (m, 2H), 7.07-7.10 (m, 2H), 6.15 (d, J = 19.2 Hz, 1H) ), 5.62-5.69 (m, 1H), 4.54-4.58 (m, 1H), 4.38-4.43 (m, 1H), 4.12-4.14 (m, 1H), 3.93-3.99 (m, 2H), 3.48-3.55 (m, 1H), 1.89-1.93 (m, 1H), 1.26-1.40 (m, 12H), 0.93-0.99 (m, 2H), 0.63-0.67 (m, 2H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.80, 3.71;

MS m/z (ESI): 586.2 [M+H] ⁺ .

Example twenty two

First step S-isopropyl (2S)-2-(((4-nitrophenoxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphino)amino Preparation of propionate

4-Nitrobenzene Phosphate (1.75 g, 7.0 mmol) was dissolved in CH ₂ Cl ₂ (15 mL), cooled to -78 ° C, 5,6,7,8-tetrahydro-2-naphthol (1.4 g, 8.0 mmol) and TEA (0.8 g, 8.0 mmol) in CH ₂ Cl ₂ (15 mL) were added dropwise at this temperature for 30 minutes, then gradually warmed to 0 ° C, and the reaction solution was added dropwise. cooled at 0 ℃ S- isopropyl (S) -2- amino-propionic acid ester hydrochloride (1.4g, 8.0mmol) in CH _₂ Cl ₂ (15mL) solution, followed by TEA (1.5g, 15mmol) was added dropwise The reaction was stirred at 0<0>C for 1 h. EtOAc was evaporated. Column chromatography (eluent: PE: EtOAc = 9:1 to 1.5:1) toield of the title compound S-isopropyl(2S)-2-(((4-nitrophenoxy)) (5,6) 7,8-tetrahydronaphthalen-2-yl)oxophosphino)amino)propanesulfate (2.2 g, 65%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.20-8.24 (m, 2H), 7.38-7.42 (m, 2H), 6.99-7.03 (m, 1H), 6.92-7.00 (m, 2H), 4.10- 4.17(m,1H), 3.79-3.82(m,2H), 3.58-3.62(m,1H), 2.72(m,4H),1.76-1.78(m,4H), 1.39(m,3H),1.24- 1.28 (m, 6H).

The second step S-isopropyl (2S)-2-(((((2(), 2R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1 (2H)) -yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphoryl) Preparation of amino)propyl sulphate

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.3 mL). ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes while stirring. After stirring at room temperature for 10 minutes, S-isopropyl (2S)-2- ((4) -Nitrophenoxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphoryl)amino)propanesulfate (480 mg, 1.0 mmol) in THF (3 mL) Instill in minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated by column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 50: 1 ~ 10: 1) to give the title compound S- isopropyl (2S) -2 - ((( ((2R, 3R, 4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) Oxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphoryl)amino)propanesulfate (148 mg, 50%, epimer ratio S _P /R _P =2.7:1).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.57-7.60 (m, 1H), 7.03-7.06 (m, 1H), 6.95-7.00 (m, 2H), 6.15 (d, J = 20.4 Hz, 1H) ), 5.57-5.64 (m, 1H), 4.55-4.59 (m, 1H), 4.38-4.45 (m, 1H), 4.12-4.14 (m, 1H), 3.93-3.99 (m, 2H), 3.51-3.56 (m, 1H), 2.74 (m, 4H), 1.80 (m, 4H), 1.27-1.39 (m, 12H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.69;

MS m/z (ESI): 600.1 [M+H] ⁺ .

Example twenty-three

First step Preparation of S-isopropyl (2S)-2-((4-nitrophenoxy)(4-(propylthio)phenoxy)phosphino)amino)propylsulfate

4-nitrophenyl phosphate dichloride (1.75g, 7.0mmol) was dissolved in CH _₂ Cl ₂ (15mL), cooled to -78 deg.] C, 4- (propylthio) phenol (1.4g, 8.0mmol) and A solution of TEA (0.8 g, 8.0 mmol) in CH ₂ Cl ₂ (15 mL) was added dropwise at this temperature for 30 minutes, then gradually warmed to 0 ° C, and the reaction solution was dropwise added to a cooled S at 0 ° C. - isopropyl (S) -2- amino-propionic acid ester hydrochloride (1.4g, 8.0mmol) in CH _₂ Cl ₂ (15mL) solution, followed by TEA (1.5g, 15mmol) was added dropwise to the reaction system at 0 ℃ The mixture was stirred for 1 hr. EtOAc was evaporated. Column chromatography (eluent: PE: EtOAc = 9:1 to 1.5:1) afforded the title compound S-isopropyl(2S)-2-(((4-nitrophenoxy)) Thio)phenoxy)phosphino)amino)propanesulfate (2.5 g, 70%).

^{_{1 H NMR (400MHz, CDCl 3}} ): δ8.21-8.24 (m, 2H), 7.37-7.41 (m, 2H), 7.291-7.32 (m, 2H), 7.14-7.18 (m, 2H), 4.10- 4.15 (m, 1H), 3.89-3.92 (m, 1H), 3.58-3.62 (m, 1H), 2.84-2.88 (m, 2H), 1.61-1.67 (m, 2H), 1.39 (m, 3H), 1.24-1.28 (m, 6H), 0.99-1.02 (m, 3H).

The second step S-isopropyl (2S)-2-(((((2(), 2R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1 (2H)) Of 4-(4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(propylthio)phenoxy)phosphoryl)amino)propanesulfate

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, at room temperature 3H)-Dione (130 mg, 0.5 mmol) was dissolved in a mixed solution of THF (5 mL) and NMP (1.3 mL). ^t BuMgCl (1.0 M in THF, 1.0 mL, 1.0 mmol) was added dropwise to the above solution over 5 minutes while stirring. After stirring at room temperature for 10 minutes, S-isopropyl (2S)-2- ((4) A solution of (4-nitrophenoxy)(4-(propylthio)phenoxy)phosphino)amino)propanesulfate (500 mg, 1.0 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 <0>C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated by column chromatography _{_{(eluent: CH 2 Cl 2: MeOH =}} 50: 1 ~ 10: 1) to give the title compound S- isopropyl (2S) -2 - ((( ((2R, 3R, 4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) (oxy)(4-(propylthio)phenoxy)phosphino)amino)propanesulfate (141 mg, 46%, epimer ratio: S _P /R _P = 1.5:1).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.61 - 7.63 (m, 1H), 7.36-7.38 (m, 2H), 7.20-7.25 (m, 2H), 6.17 (d, J = 17.2 Hz, 1H) ), 5.66-5.71 (m, 1H), 4.55-4.59 (m, 1H), 4.40-4.45 (m, 1H), 4.12-4.15 (m, 1H), 3.94-4.00 (m, 2H), 3.48-3.58 (m, 1H), 2.88-2.92 (m, 2H), 1.60-1.67 (m, 2H), 1.26-1.40 (m, 12H), 1.00-1.05 (m, 3H);

³¹ P NMR (162 MHz, CD ₃ OD): δ 3.70;

MS m/z (ESI): 620.2 [M+H] ⁺ .

Example twenty four

First step Preparation of 4-cyclopropylphenylphosphorus dichloride

4-Cyclopropylphenol (2.6 g, 19.39 mmol) was weighed and dissolved in Et ₂ O (15 mL). TEA (2.7 mL, 19.39 mmol) was slowly added dropwise at room temperature and stirred for 15 minutes.

Phosphorus oxychloride (3.0 g, 19.39 mmol) was dissolved in Et ₂ O (100 mL), and after cooling to -55 ° C, the solution was slowly added dropwise, and a white solid was precipitated. After the completion of the dropwise addition, stirring was continued at this temperature for 2 hours, then slowly warmed to room temperature and stirred overnight. The white solid was filtered off under N _2, and the filtrate was concentrated to give the title compound 4-cyclopropyl-phenyl phosphoric acid ester dichloride (4.4g, 91%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ7.18-7.14 (m, 2H), 7.12-7.08 (m, 2H), 1.94-1.86 (m, 1H), 1.03-0.96 (m, 2H), 0.71-0.65 (m, 2H);

³¹ P NMR (162 MHz, CDCl ₃ ) δ 3.99.

Second step Preparation of isopropyl ((4-cyclopropylphenoxy)(perfluorophenoxy)phosphoryl)-L-alanine ester

L-Alanine isopropyl ester hydrochloride (2.5 g, 14.97 mmol) was dissolved in CH ₂ Cl ₂ (30 mL), cooled to -78 ° C, and TEA (4.3 mL, 31.44) was slowly added dropwise under N ₂ protection. After stirring for 15 minutes, a solution of 4-cyclopropylphenylphosphoric dichloride (3.7 g, 14.82 mmol) dissolved in CH ₂ Cl ₂ (5 mL) was slowly added dropwise, and then stirred slowly for 30 minutes. Stirring was continued at this temperature for 1 hour at 0 °C.

The perfluorophenol (2.72 g, 14.82 mmol) was dissolved in CH ₂ Cl ₂ (10 mL), and TEA (2.3 mL, 16.46 mmol) was slowly added dropwise at 0 ° C. After stirring for 5 minutes, the solution was slowly added dropwise to the above reaction. In the system, keep 0 ° C and stir overnight. The solid was filtered off, and the filtrate was evaporated, evaporated, mjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Phospho)-L-alanine ester (6.03 g, 82%, epimer ratio: S _P /R _P = 1:1).

^{1 H NMR (400MHz, DMSO-} d 6) δ7.16-7.08 (m, 4H), 6.80-6.73 (m, 1H), 4.91-4.84 (m, 1H), 3.97-3.87 (m, 1H), 1.95 -1.89 (m, 1H), 1.33-1.23 (m, 3H), 1.18-1.14 (m, 6H), 0.96-0.90 (m, 2H), 0.65-0.59 (m, 2H);

³¹ P NMR (162 MHz, DMSO-d ₆ ) δ 0.52.

Third step Preparation of isopropyl ((S)-(4-cyclopropylphenoxy)(perfluorophenoxy)phosphoryl)-L-alanine ester

The isopropyl ((4-cyclopropylphenoxy)(perfluorophenoxy)phosphino)-L-alanine ester obtained in the second step above (the ratio of epimers is S _P /R _P The mixture was filtered with EtOAc (EtOAc) = EtOAc (EtOAc) (Perfluorophenoxy)phosphino)-L-alanine ester (2.99 g, 40%, HPLC purity: 97%).

^{1 H NMR (400MHz, DMSO-} d 6) δ7.09 (s, 4H), 6.80 (dd, J = 14.0,10.0Hz, 1H), 4.89-4.83 (m, 1H), 3.96-3.87 (m, 1H ), 1.95-1.89 (m, 1H), 1.26 (d, J = 6.8 Hz, 3H), 1.16-1.14 (m, 6H), 0.96-0.90 (m, 2H), 0.65-0.60 (m, 2H);

³¹ P NMR (162 MHz, DMSO-d ₆ ) δ 0.52.

The fourth step is isopropyl (S)-(4-cyclopropylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidine- Preparation of 1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine ester

Take 1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H) - dione (440mg, 1.69mmol) was dissolved in THF (10mL), cooled to -5 deg.] C, was slowly added dropwise at ^t BuMgCl under N ₂ (1M in THF, 3.6mL, 3.6mmol ), after the addition was complete Stir at this temperature for 30 minutes, then warm to room temperature and continue stirring for 30 minutes. The reaction system was placed in an ice water bath, and isopropyl ((S)-(4-cyclopropylphenoxy)(perfluorophenoxy)phosphino) dissolved in THF (3.6 mL) was slowly added dropwise. A solution of L-alanine ester (1.0 g, 2.03 mmol) was maintained at this temperature and stirring was continued overnight. With 1N HCl solution (2mL) was quenched reaction was diluted with CH _₂ Cl ₂ (50mL) and a 1N HCl solution (20mL) residue was added after concentration, organic phase was separated, the aqueous phase was _{_{CH 2 Cl 2 (50mL × 2}} ) and extracted with, The combined organic layers were washed with EtOAc EtOAc EtOAc. The resulting solid was recrystallized from CH _₂ Cl _2, to give the title compound isopropyl (S) - (4- cyclopropylphenoxy) (((2R, 3R, 4R, 5R) -5- (2,4 -dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine Acid ester (680 mg, 70%, HPLC purity: 98%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ9.25 (brs, 1H), 7.48 (d, J = 7.6,1H), 7.11-7.08 (m, 2H), 7.03-7.01 (m, 2H), 6.18 (d , J = 18.4 Hz, 1H), 5.70 (d, J = 8.0, 1H), 5.03-4.99 (m, 1H), 4.55 - 4.43 (m, 2H), 4.13-4.08 (m, 2H), 3.96-3.91 (m, 2H), 1.90-8.11 (m, 1H), 1.45-1.32 (m, 6H), 1.31-1.19 (m, 6H), 0.98-0.91 (m, 2H), 0.66-0.61 (m, 2H) ;

³¹ P NMR (162 MHz, CDCl ₃ ) δ 3.65;

¹⁹ F NMR (376 MHz, CDCl ₃ ) δ-161.79;

MS m/z (ESI): 570.1 [M+H] ⁺ .

Isopropyl ((R)-(4-cyclopropylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidine-1 ( Preparation of 2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine ester

Take 1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H) - dione (220mg, 0.845mmol) was dissolved in THF (5mL), cooled to -5 deg.] C, was slowly added dropwise at ^t BuMgCl under N ₂ (1M in THF, 1.8mL, 1.8mmol ), after the addition was complete Stir at this temperature for 30 minutes, then warm to room temperature and continue stirring for 30 minutes. The reaction system was placed in an ice water bath, and isopropyl ((4-cyclopropylphenoxy)(perfluorophenoxy)phosphino)-L-alanine dissolved in THF (3 mL) was slowly added dropwise. The ester (second step in Example 21) (500 mg, 1.02 mmol) was maintained at this temperature and stirred continuously overnight. With 1N HCl solution (1 mL) quenched the reaction system, the solvent was concentrated under reduced pressure pump, (15 mL) was added the residue diluted with CH _₂ Cl ₂ (30mL) and a 1N HCl solution, organic phase was separated, the aqueous phase was _extracted with ₂ CH Cl (30mL The organic phase was combined, washed with a saturated aqueous sodium hydrogen carbonate solution (20 mL) and brine (20 mL). Column chromatography (CH ₂ Cl ₂ : ⁱ Pr OH = 20:1) gave the title compound isopropyl ((R)-(4-cyclopropylphenoxy) (((2R,3R,4R,5R) -5-(2,4-Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy) Phosphoryl)-L-alanine ester (118 mg, 24%, HPLC purity: 98%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ8.95 (s, 1H), 7.26-7.24 (m, 1H), 7.11-7.09 (m, 2H), 7.03-7.00 (m, 2H), 6.17 (d, J =18.8 Hz, 1H), 5.59 (d, J = 8.0, 1H), 5.05-4.99 (m, 1H), 4.51-4.45 (m, 2H), 4.10 (d, J = 9.6 Hz, 1H), 4.06- 3.88 (m, 2H), 3.76 (dd, J = 24.0, 9.2, 1H), 1.87-1.82 (m, 1H), 1.39-1.28 (m, 6H), 1.27-1.21 (m, 6H), 0.98-0.91 (m, 2H), 0.66-0.55 (m, 2H);

³¹ P NMR (162 MHz, CDCl ₃ ) δ 4.31;

¹⁹ F NMR (376 MHz, CDCl ₃ ) δ -162.04;

MS m/z (ESI): 570.1 [M+H] ⁺ .

Example twenty five

Isopropyl ((S)-(((2R,3R,4R,5R)-3-acetoxy-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl) -4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)(4-cyclopropylphenoxy)phosphino)-L-alaninate

Take isopropyl (S)-(4-cyclopropylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1) 2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphino)-L-alaninate (57 mg, 0.10 mmol) was dissolved in THF (7 mL) To the solution, acetyl chloride (0.035 mL, 0.50 mmol), DMAP (61 mg, 0.50 mmol) was sequentially added, and the mixture was stirred at room temperature for 3 hours. Diluted with EtOAc, the organic phase washed successively with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated, Prep TLC-purified (eluent _{_{^{CH 2 Cl 2: i PrOH =}}} 20: 1), to give the title compound isopropyl ((S)-((2R,3R,4R,5R)-3-acetoxy-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4 -Fluoro-4-methyltetrahydrofuran-2-yl)methoxy)(4-cyclopropylphenoxy)phosphino)-L-alaninate (45 mg, 74%, HPLC purity: 98%)

^{1 H NMR (400MHz, MeOD)} δ7.55 (d, J = 8.0Hz, 1H), 7.06 (m, 4H), 6.03 (d, J = 18.4Hz, 1H), 5.55 (d, J = 8.0Hz, 1H), 5.22 (m, 1H), 4.91 (m, 1H), 4.38 (m, 1H), 4.27 (m, 2H), 3.85 (m, 1H), 3.26 (m, 1H), 2.10 (s, 3H) ), 1.83 (m, 1H), 1.30 (m, 6H), 1.17 (m, 6H), 0.89 (m, 2H), 0.58 (m, 2H);

³¹ P NMR (162 MHz, MeOD) δ 3.83;

¹⁹ F NMR (376 MHz, MeOD) δ-157.42;

MS m/z (ESI): 612.2 [M+H] ⁺ .

Example twenty six

First step isopropyl ((S)-((2,3-dihydro-1H-indol-5-yl)oxy)(perfluorophenoxy)phosphino)-L-alaninate preparation

2,3-Dihydro-1H-indol-5-ol (20.0 g, 150 mmol) was weighed and dissolved in Et ₂ O (150 mL). DIPEA (19.4 g, 150 mmol) was slowly added dropwise at room temperature and stirred for 15 minutes.

Phosphorus oxychloride (23.0 g, 150 mmol) was dissolved in Et ₂ O (300 mL), and the solution was slowly added dropwise to -78 ° C, and the solution was slowly added dropwise over 1.5 hours, and a white solid was precipitated. After the addition was completed, it was slowly warmed to room temperature and stirred overnight. LCMS showed the reaction to give (2,3-dihydro-1H-indol-5-yl)phosphoric dichloride.

L-Alanine isopropyl ester hydrochloride (24.0 g, 143 mmol) was dissolved in CH ₂ Cl ₂ (400 mL), and DIPEA (39.0 g, 300 mmol) was added and stirred well. The solution was slowly added dropwise to a (2,3-dihydro-1H-indol-5-yl)phosphoric dichloride solution cooled at -78 ° C for about 2 hours. The solution slowly rose to 0 °C.

The perfluorophenol (30.3 g, 165 mmol) was weighed and dissolved in CH ₂ Cl ₂ (100 mL). The solution was placed in an ice bath, and DIPEA (21.5 g, 165 mmol) was slowly added dropwise with stirring. The above solution was slowly dropped into a zero-degree solution to which L-alanine isopropyl ester hydrochloride had been added, and the solution was allowed to stand for half an hour, and the solution was stirred at room temperature overnight.

The solution was washed with water (2 L), EtOAc (EtOAc) (EtOAc)

Petroleum ether (1.25 L), EtOAc (40 mL). The solution was filtered, washed with EtOAc (EtOAc) (EtOAc) (Perfluorophenoxy)phosphino)-L-alaninate (14.5 g, 20%).

^{1 H NMR (400MHz, MeOD)} δ7.20 (d, J = 8.4Hz, 1H), 7.12 (s, 1H), 7.01 (d, J = 8.4Hz, 1H), 4.97 (m, 1H), 4.02 ( m, 1H), 2.90 (m, 4H), 2.12 (m, 2H), 1.38 (dd, J = 7.2, 1.0 Hz, 3H), 1.23 (m, 6H);

¹⁹ F NMR (376 MHz, MeOD) δ - 155.35 (d, J = 22.4 Hz, 2F), -163.11 (t, J = 23.2 Hz, 1F), -165.78 (t, J = 23.2 Hz, 3F).

The second step is isopropyl (S)-((2,3-dihydro-1H-indol-5-yl)oxo)(((2R,3R,4R,5R)-5-(2,4-di) Carbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine Preparation of ester

Take 1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H) - dione (2.95g, 11.36mmol) was dissolved in THF (100mL), cooled to 0 ℃, in slowly added dropwise under N ₂ ^{t BuMgCl (1M in THF, 22.72mL} , 22.72mmol), after the addition was complete Stir at this temperature for 30 minutes, then warm to room temperature and continue stirring for 30 minutes. The reaction system was placed in an ice water bath, and isopropyl ((S)-((2,3-dihydro-1H-indol-5-yl)oxy) dissolved in THF (50 mL) was slowly added dropwise (all) A solution of fluorophenoxy)phosphino)-L-alaninate (5.6 g, 11.36 mmol) was maintained at this temperature and stirred continuously overnight. LCMS showed the product main peak. Solution was washed with 1N HCl solution (20mL) was quenched reaction was diluted with CH _₂ Cl ₂ (300mL) and a 1N HCl solution (100 mL) The residue was added after concentration, organic phase was separated, the aqueous phase was _{_{CH 2 Cl 2 (50mL × 2}} ) and extracted with The combined organic layers were washed with aq. EtOAc (EtOAc) Column chromatography (eluent: CH ₂ Cl ₂ : ⁱ Pr OH = 60:1 to 12:1) to give the title compound isopropyl(S)-((2,3-dihydro-1H-indole-5-yl) Oxo)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- 4-Methyltetrahydrofuran-2-yl)methoxy)phosphino)-L-alaninate (3.9 g, 67%).

^{1 H NMR (400MHz, MeOD)} 7.61 (d, J = 8.0Hz, 1H), 7.18 (d, J = 8.0Hz, 1H), 7.12 (s, 1H), 7.01 (dd, J = 8.4Hz, 0.8Hz , 1H), 6.14 (d, J = 19.6 Hz, 1H), 5.57 (d, J = 8.4 Hz, 1H), 4.98 (m, 1H), 4.54 (m, 1H), 4.38 (m, 1H), 4.12 (m, 1H), 3.95 (m, 2H), 2.88 (m, 4H), 2.10 (m, 2H), 1.39-1.33 (m, 6H), 1.24 (m, 6H);

³¹ P NMR (162 MHz, MeOD) δ 3.89;

¹⁹ F NMR (376 MHz, CDCl ₃ ) δ-161.97;

MS m/z (ESI): 570.1 [M+H] ⁺ .

Example twenty seven

Take isopropyl (S)-((2,3-dihydro-1H-indol-5-yl)oxo)((2R,3R,4R,5R)-5-(2,4-dicarbonyl- 3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alaninate 57 mg, 0.10 mmol) was dissolved in THF (7 mL). EtOAc (EtOAc (EtOAc) Diluted with EtOAc, the organic phase washed successively with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated, Prep TLC-purified (eluent _{_{^{CH 2 Cl 2: i PrOH =}}} 20: 1), to give the title compound isopropyl ((S)-((2R,3R,4R,5R)-3-acetoxy-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4 -Fluoro-4-methyltetrahydrofuran-2-yl)methoxy)(4-cyclopropylphenoxy)phosphino)-L-alaninate (40 mg, 65%).

^{1 H NMR (400MHz, CDCl3)} δ9.37 (s, 1H), 7.48 (d, J = 8.0Hz, 1H), 7.06 (d, J = 8.0Hz, 1H), 7.01 (s, 1H), 6.88 ( d, J = 8.0 Hz, 1H), 6.13 (d, J = 18.4 Hz, 1H), 5.46 (d, J = 8.0 Hz, 1H), 5.12 (dd, J = 20.4, 8.0 Hz, 1H), 4.94 ( m, 1H), 4.47 (m, 1H), 4.17 (m, 1H), 3.90 (m, 2H), 2.78 (m, 4H), 2.11 (s, 3H), 2.01 (m, 2H), 1.29 (m) , 6H), 1.16 (m, 6H);

³¹ P NMR (162 MHz, CDCl ₃ ) δ 2.97;

¹⁹ F NMR (376 MHz, CDCl ₃ ) δ - 157.52;

MS m/z (ESI): 612.3 [M+H] ⁺ .

Example twenty eight

First step isopropyl ((S)-((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)(perfluorophenoxy)phosphino)-L-alanine Preparation

5,6,7,8-tetrahydronaphthalen-2-ol (23.0 g, 150 mmol) was weighed and dissolved in Et ₂ O (150 mL). DIPEA (19.4 g, 150 mmol) was slowly added dropwise at room temperature and stirred for 15 minutes.

Phosphorus oxychloride (23.0 g, 150 mmol) was dissolved in Et ₂ O (300 mL), and the solution was slowly added dropwise to -78 ° C, and the solution was slowly added dropwise over 1.5 hours, and a white solid was precipitated. After the completion of the dropwise addition, the mixture was slowly warmed to room temperature and stirred overnight, and LCMS showed to give (5,6,7,8-tetrahydronaphthalen-2-yl)phosphorous dichloride.

L-Alanine isopropyl ester hydrochloride (24.0 g, 143 mmol) was dissolved in CH ₂ Cl ₂ (400 mL), and DIPEA (39.0 g, 300 mmol) was added and stirred well. The solution of (5,6,7,8-tetrahydronaphthalen-2-yl)phosphoric dichloride ester which was cooled at -78 ° C was slowly dropped, and the dropwise addition time was about 2 hours. The solution slowly rose to zero degrees.

The solution was washed with water (2 mL), EtOAc (EtOAc)

Petroleum ether (500 mL), ethyl acetate (15 mL) was added to the obtained mixture, and the suspension was stirred at room temperature for 3 hours and cooled to 0 ° C, and stirred at 0 ° C for 1 hour. The solution was filtered, washed with EtOAc (50 mL) EtOAc. ¹⁹ F NMR in MeOD showed a 4:1 epimer. The solid was stirred further in petroleum ether (250 mL) EtOAc (EtOAc)EtOAc. The solid was further stirred in petroleum ether (300 mL), EtOAc (EtOAc) (EtOAcjjjjjjjj Oxo)(perfluorophenoxy)phosphino)-L-alaninate (22.0 g, 28%).

¹ H NMR (400 MHz, MeOD) δ 7.06 (d, J = 9.2 Hz, 1H), 6.97 (m, 2H), 4.95 (m, 1H), 4.02 (m, 1H), 2.76 (m, 4H), 1.81 (m, 4H), 1.39 (dd, J = 7.2, 1.2 Hz, 3H), 1.23 (m, 6H);

¹⁹ F NMR (376 MHz, MeOD) δ-155.33 (d, J = 18.8 Hz, 2F), -163.14 (t, J = 23.2 Hz, 1F), -165.80 (t, J = 20.2 Hz, 3F).

The second step is isopropyl (S)-((5,6,7,8-tetrahydronaphthalen-2-yl)oxo)(((2R,3R,4R,5R)-5-(2,4- Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine Preparation of acid ester

Take 1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H) - dione (2.95g, 11.36mmol) was dissolved in THF (100mL), cooled to 0 ℃, in slowly added dropwise under N ₂ ^{t BuMgCl (1M in THF, 22.72mL} , 22.72mmol), after the addition was complete Stir at this temperature for 30 minutes, then warm to room temperature and continue stirring for 30 minutes. The reaction system was placed in an ice water bath, and isopropyl ((S)-((5,6,7,8-tetrahydronaphthalen-2-yl)oxy) dissolved in THF (50 mL) was slowly added dropwise ( A solution of perfluorophenoxy)phosphino)-L-alaninate (5.8 g, 11.36 mmol) was maintained at this temperature and stirred continuously overnight. Reaction with 1N HCl solution (20mL) quenched after concentration the residue was added CH _₂ Cl ₂ (300mL) and a 1N HCl solution (100 mL) was diluted organic phase was separated, the aqueous phase was extracted with _{_{CH 2 Cl 2 (50mL × 2}} ), combined the organic phase were saturated sodium bicarbonate solution (100 mL) and saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated by column chromatography _{_{^{(eluent: CH 2 Cl 2: i PrOH}}} = 60: 1~12:1) The title compound isopropyl(S)-((5,6,7,8-tetrahydronaphthalen-2-yl)oxo)((2R,3R,4R,5R)-5 -(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl) -L-Alanine ester (2.2 g, 34%).

^{1 H NMR (400MHz, MeOD)} 7.59 (d, J = 7.2Hz, 1H), 7.04 (m, 1H), 6.97 (m, 1H), 6.14 (d, J = 18.8Hz, 1H), 5.55 (d, J=8.0 Hz, 1H), 4.96 (m, 1H), 4.55 (m, 1H), 4.38 (m, 1H), 4.12 (m, 1H), 3.94 (m, 2H), 2.74 (m, 4H), 1.79 (m, 4H), 1.39-1.33 (m, 6H), 1.24 (m, 6H);

³¹ P NMR (162 MHz, MeOD) δ 3.87;

¹⁹ F NMR (376 MHz, MeOD) δ -162.11;

MS m/z (ESI): 584.0 [M+H] ⁺ .

Example twenty nine

Isopropyl ((S)-(((2R,3R,4R,5R)-3-acetoxy-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl) -4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphoryl)-L-alanine Acid ester

Take isopropyl (S)-((5,6,7,8-tetrahydronaphthalen-2-yl)oxo)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl) -3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine (60 mg, 0.10 mmol) was dissolved in THF (7 mL). EtOAc (EtOAc (EtOAc,EtOAc) . Diluted with EtOAc, the organic phase washed successively with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated by column chromatography _{_{^{(eluent: CH 2 Cl 2: i PrOH}}} = 60: 1 ~ 12: 1) to give the title The compound isopropyl ((S)-((2R,3R,4R,5R)-3-acetoxy-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-) 4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphoryl)-L-alanine Acid ester (29 mg, 40%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ9.14 (s, 1H), 7.54 (d, J = 8.4Hz, 1H), 6.99 (m, 1H), 6.92 (m, 2H), 6.20 (d, J = 18.4 Hz, 1H), 5.51 (d, J = 8.0 Hz, 1H), 5.17 (dd, J = 20.4, 9.2 Hz, 2H), 5.00 (m, 1H), 4.55 (m, 1H), 4.31 (m, 1H), 4.22 (m, 1H), 3.92 (m, 2H), 2.70 (m, 4H), 2.17 (s, 3H), 1.76 (m, 4H), 1.34 (m, 6H), 1.24 (m, 6H) );

³¹ P NMR (162 MHz, CDCl ₃ ) δ 2.87;

¹⁹ F NMR (376 MHz, CDCl ₃ ) δ - 157.54;

MS m/z (ESI): 626.3 [M+H] ⁺ .

Example thirty

First step Preparation of isopropyl ((S)-(4-(propylthio)phenoxy)(perfluorophenoxy)phosphino)-L-alanine ester

4-(propylthio)phenol (14.7 g, 87.5 mmol) was weighed and dissolved in Et ₂ O (100 mL). DIPEA (11.3 g, 87.5 mmol) was slowly added dropwise at room temperature and stirred for 15 minutes.

Phosphorus oxychloride (14.0 g, 91.9 mmol) was dissolved in Et ₂ O (200 mL), and the solution was slowly added dropwise to -78 ° C, and the solution was slowly added dropwise over 1.5 hours, and a white solid was precipitated. After the completion of the dropwise addition, the temperature was slowly raised to room temperature and stirred overnight, and LCMS showed to give (4-(propylthio)phenoxy)phosphoric acid dichloride.

L-Alanine isopropyl ester hydrochloride (14.0 g, 83.8 mmol) was dissolved in CH ₂ Cl ₂ (180 mL), and DIPEA (22.8 g, 177 mmol) was added and stirred well. The solution of (4-(propylthio)phenoxy)phosphoric dichloride ester which was cooled at -78 ° C was slowly added dropwise for about 2 hours, and the solution was slowly raised to 0 ° C after the completion of the dropwise addition.

The perfluorophenol (17.7 g, 96 mmol) was weighed and dissolved in CH ₂ Cl ₂ (120 mL), and the solution was placed in an ice bath, and DIPEA (12.4 g, 96 mmol) was slowly added dropwise with stirring. The above solution was slowly dropped into a solution at 0 ° C to which L-alanine isopropyl ester hydrochloride had been added, and the solution was allowed to drip for half an hour, and the solution was stirred at room temperature overnight.

The solution was washed with water (1 L), EtOAc (EtOAc)

Petroleum ether (650 mL), ethyl acetate (130 mL) was added, and the suspension was stirred at room temperature overnight. The solution was filtered, washed with EtOAc (EtOAc) (EtOAc) (EtOAc) Acid ester (10.9 g, 27%).

^{1 H NMR (400MHz, MeOD)} δ7.39 (m, 2H), 7.22 (m, 2H), 4.97 (m, 1H), 4.03 (m, 1H), 2.92 (t, J = 7.2Hz, 2H), 1.84 (m, 2H), 1.39 (dd, J = 7.2, 1.2 Hz, 3H), 1.23 (m, 6H), 1.03 (t, J = 7.2 Hz, 3H);

¹⁹ F NMR (376 MHz, MeOD) δ - 155.40 (d, J = 22.0 Hz, 2F), -162.95 (t, J = 22.0 Hz, 1F), -165.78 (t, J = 20.2 Hz, 3F).

The second step is isopropyl (S)-(4-(propylthio)phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-di) Preparation of Hydropyrimidine-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine

Take 1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H) - dione (1.56g, 6.0mmol) was dissolved in THF (50mL), cooled to 0 ℃, in slowly added dropwise under N ₂ ^{t BuMgCl (1M in THF, 12.0mL} , 12.0mmol), after the addition was complete Stir at this temperature for 30 minutes, then warm to room temperature and continue stirring for 30 minutes. The reaction system was placed in an ice water bath, and isopropyl ((S)-(4-(propylthio))phenoxy) (perfluorophenoxy)phosphino group dissolved in THF (25 mL) was slowly added dropwise. A solution of -L-alanine ester (3.32 g, 6.3 mmol) was maintained at this temperature and stirred continuously overnight. LCMS showed the product main peak. Solution was washed with 1N HCl solution (12 mL) quenched the reaction system, diluted with CH _₂ Cl ₂ (150mL) and a 1N HCl solution (50mL) residue was added after concentration, organic phase was separated, the aqueous phase was _{_{CH 2 Cl 2 (25mL × 2}} ) and extracted with The combined organic phases were saturated sodium bicarbonate solution (50mL) and brine (50mL), dried over anhydrous sodium sulfate, filtered, column chromatography _{_{^{(eluent: CH 2 Cl 2: i PrOH}}} = 60: 1-12:1) The title compound isopropyl(S)-(4-(propylthio)phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl) -3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)-L-alanine (1.9g, 53%).

^{1 H NMR (400MHz, MeOD)} 7.63 (d, J = 8.4Hz, 1H), 7.36 (d, J = 8.4Hz, 1H), 7.22 (dd, J = 8.8,0.8Hz, 2H), 6.16 (d, J = 20.0 Hz, 1H), 5.66 (d, J = 8.0 Hz, 1H), 4.96 (m, 1H), 4.55 (m, 1H), 4.40 (m, 1H), 4.14 (m, 1H), 3.95 ( m, 2H), 2.90 (t, J = 6.8 Hz, 2H), 1.64 (m, 2H), 1.39-1.33 (m, 6H), 1.24 (m, 6H), 1.03 (t, J = 8.0 Hz, 3H) );

³¹ P NMR (162 MHz, MeOD) δ 3.89;

¹⁹ F NMR (376 MHz, MeOD) δ-161.87;

MS m/z (ESI): 604.0 [M+H] ⁺ .

Embodiment 31

Isopropyl ((S)-(((2R,3R,4R,5R)-3-acetoxy-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl) -4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)(4-(propylthio)phenoxy)phosphino)-L-alaninate

Take isopropyl (S)-(4-(propylthio)phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidine) -1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphino)-L-alaninate (60 mg, 0.10 mmol) was dissolved To the solution were added acetyl chloride (0.035 mL, 0.50 mmol), EtOAc (EtOAc) Diluted with EtOAc, the organic phase washed successively with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated, Prep TLC-purified (eluent _{_{^{CH 2 Cl 2: i PrOH =}}} 20: 1), to give the title compound isopropyl ((S)-((2R,3R,4R,5R)-3-acetoxy-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4 -Fluoro-4-methyltetrahydrofuran-2-yl)methoxy)(4-(propylthio)phenoxy)phosphino)-L-alaninate (50 mg, 77%).

^{1 H NMR (400MHz, MeOD)} δ7.53 (d, J = 8.0Hz, 1H), 7.25 (dd, J = 11.6,2.8Hz, 2H), 7.10 (dd, J = 8.8,1.2Hz, 2H), 6.01 (d, J = 20.4 Hz, 1H), 5.54 (d, J = 8.8 Hz, 1H), 5.17 (dd, J = 20.4, 8.0 Hz, 2H), 4.85 (m, 1H), 4.38 (m, 1H) ), 4.22 (m, 2H), 3.83 (m, 1H), 2.78 (t, J = 7.2 Hz, 2H), 1.50 (m, 2H), 1.26 (m, 6H), 1.17 (m, 6H), 0.96 (t, J = 7.2 Hz, 3H);

³¹ P NMR (162 MHz, MeOD) δ 3.85;

¹⁹ F NMR (376 MHz, MeOD) δ-157.38;

MS m/z (ESI): 646.2 [M+H] ⁺ .

Example thirty-two

First step S-isopropyl (S)-2-(((S)-(perfluorophenoxy)(phenoxy)phosphino)amino)propyl sulfate

Phenol (9.4 g, 100 mmol) was weighed and dissolved in Et ₂ O (100 mL). DIPEA (12.9 g, 100 mmol) was slowly added dropwise at room temperature and stirred for 15 minutes.

Phosphorus oxychloride (16.0 g, 105 mmol) was dissolved in Et ₂ O (200 mL). After cooling to -78 ° C, the solution was slowly added dropwise. After 1.5 hours, a white solid was precipitated. After stirring at room temperature overnight, LCMS showed the formation of phenylphosphorous dichloride.

S-Isopropyl (S)-2-aminopropanesulfate hydrochloride (16.5 g, 90 mmol) was dissolved in CH ₂ Cl ₂ (200 mL), and DIPEA (23.2 g, 180 mmol) was added and stirred. The solution was slowly dropped into a cooled phenylphosphoric dichloride solution at -78 ° C for about 2 hours. The solution slowly rose to 0 °C.

The perfluorophenol (20.2 g, 110 mmol) was weighed and dissolved in CH ₂ Cl ₂ (100 mL), and the solution was placed in an ice bath, and DIPEA (14.2 g, 110 mmol) was slowly added dropwise with stirring. The above solution was slowly dropped into a zero-degree solution to which S-isopropyl(S)-2-aminopropylsulfate hydrochloride had been added, and the mixture was stirred for 30 minutes, and the solution was stirred at room temperature overnight.

The solution was washed with water (1 L), EtOAc (EtOAc)

Petroleum ether (300 mL), EtOAc (30 mL). The solution was filtered, washed with petroleum ether (50 mL), and the solvent was evaporated on a vacuum apparatus to give 13.5 g of crude product containing a pair of epimers. The product was beaten five times with a mixed solvent (PE: EtOAc = 20:1) The title compound S-isopropyl(S)-2-(((S)-(perfluorophenoxy)(phenoxy)phosphino)amino)propylsulfate (7.1 g, 15%) was obtained.

^{1 H NMR (400MHz, MeOD)} δ7.42 (m, 2H), 7.28 (m, 3H), 4.08 (m, 1H), 3.52 (m, 1H), 1.38 (dd, J = 7.2,0.8Hz, 3H ), 1.26 (m, 6H);

¹⁹ F NMR (376 MHz, MeOD) δ-154.86 (d, J = 20.0 Hz, 2F), -16.92 (t, J = 21.2 Hz, 1F), -165.65 (t, J = 18.4 Hz, 3F);

³¹ P NMR (162 MHz, MeOD) δ 0.07.

The second step S-isopropyl (S)-2-(((S)-(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidine- Preparation of 1(2H)-yl)-4-fluoro-3-hydroxy-4-methylhydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propanesulfate

Take 1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H) - dione (1.56g, 6.0mmol) was dissolved in THF (50mL), cooled to 0 ℃, in slowly added dropwise under N ₂ ^{t BuMgCl (1M in THF, 12.0mL} , 12.0mmol), after the addition was complete Stir at this temperature for 30 minutes, then warm to room temperature and continue stirring for 30 minutes. The reaction system was placed in an ice water bath, and S-isopropyl (S)-2-(((S)-(perfluorophenoxy)(phenoxy)phosphino)amino)propyl sulfate was slowly added dropwise ( A solution of 2.96 g, 6.3 mmol) in THF (50 mL) Reaction with 1N HCl solution (12 mL) The reaction was quenched, diluted with CH _₂ Cl ₂ (150mL) and a 1N HCl solution (30mL) residue was added after concentration, organic phase was separated, the aqueous phase was _{_{CH 2 Cl 2 (25mL × 2}} ) and extracted with, the combined organic phases were saturated sodium bicarbonate solution (30mL) and brine (30mL), dried over anhydrous sodium sulfate, filtered and concentrated by column chromatography _{_{^{(eluent: CH 2 Cl 2: i PrOH}}} = 60: 1 to 12:1) 0.93 g of crude product was obtained, which was crystallized from EtOAc (EtOAc) -5-(2,4-Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methylhydrofuran-2-yl)methoxy (Phenoxy)phosphoryl)amino)propylsulfate (0.5 g, 15%).

^{1 H NMR (400MHz, MeOD)} 7.63 (d, J = 8.0Hz, 1H), 7.40 (m, 2H), 7.28 (m, 2H), 7.21 (m, 1H), 6.15 (d, J = 20.0Hz, 1H), 5.65 (d, J = 8.0 Hz, 1H), 4.57 (m, 1H), 4.42 (m, 1H), 4.13 (m, 1H), 3.98 (m, 2H), 3.52 (m, 1H), 1.39-1.33 (m, 6H), 1.27 (m, 6H);

³¹ P NMR (162 MHz, MeOD) δ 3.53;

¹⁹ F NMR (376 MHz, MeOD) δ -161.97;

MS m/z (ESI): 546.0 [M+H] ⁺ .

Example thirty three

(2R,3R,4R,5R)-5-(2,4-Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-2-(((())) ((S)-1-(Isopropylthio)-1-carbonylpropan-2-yl)amino)(phenoxy)phosphoryl)oxo)methyl)-4-methyltetrahydrofuran-3-yl Acetate

Take S-isopropyl (S)-2-(((S)-(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1) 2H)-yl)-4-fluoro-3-hydroxy-4-methylhydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propanesulfate (54 mg, 0.10 mmol) To the solution were added acetyl chloride (0.035 mL, 0.50 mmol), EtOAc (EtOAc) Diluted with EtOAc, the organic phase washed successively with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated by column chromatography _{_{^{(eluent: CH 2 Cl 2: i PrOH}}} = 60: 1 ~ 12: 1) to give the title Compound (2R,3R,4R,5R)-5-(2,4-Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-2-((((S)) (((S)-1-(isopropylthio)-1-carbonylpropan-2-yl)amino)(phenoxy)phosphoryl)oxo)methyl)-4-methyltetrahydrofuran-3- Acetate (52 mg, 90%).

^{_{1 H NMR (400MHz, CDCl 3}} ) δ8.92 (s, 1H), 7.49 (d, J = 8.0Hz, 1H), 7.34 (m, 2H), 7.19 (m, 3H), 6.18 (d, J = 18.8 Hz, 1H), 5.50 (m, 1H), 5.18 (dd, J = 20.8, 9.2 Hz, 1H), 4.56 (m, 1H), 4.31 (m, 1H), 4.24 (m, 1H), 4.06 ( m, 1H), 3.96 (m, 1H), 3.61 (m, 1H), 2.19 (s, 3H), 1.39-1.33 (m, 6H), 1.28 (m, 6H);

³¹ P NMR (162 MHz, CDCl ₃ ) δ 2.36;

¹⁹ F NMR (376 MHz, CDCl ₃ ) δ-157.20;

MS m/z (ESI): 588.0 [M+H] ⁺ .

Embodiment thirty four

First step Preparation of S-isopropyl (2S)-2-((4-cyclopropylphenoxy)(perfluorophenoxy)phosphoryl)amino)propanesulfate

4-Cyclopropylphenol (4.4 g, 32.8 mmol) was weighed and dissolved in Et ₂ O (30 mL), and DIPEA (4.23 g, 32.8 mmol) was slowly added dropwise to the reaction system at room temperature and stirred for 15 minutes.

Phosphorus oxychloride (5.0 g, 32.8 mmol) was dissolved in Et ₂ O (60 mL), and after cooling to -78 ° C, the above solution was slowly added dropwise, and the mixture was dropped over 1.5 hours, and a white solid precipitated. After the addition was completed, it was slowly warmed to room temperature and stirred overnight. LCMS showed the formation of 4-cyclopropylphenylphosphonium dichloride.

Take S- isopropyl (S) -2- Amino-propionic acid ester hydrochloride (5.67g, 31mmol) was dissolved in CH _₂ Cl ₂ (80mL), was added DIPEA (8.5g, 66mmol), stir. The solution was slowly dropped into a cooled phenylphosphoric dichloride solution at -78 ° C for about 2 hours, and the solution was slowly warmed to 0 ° C after the completion of the dropwise addition.

The perfluorophenol (6.62 g, 36 mmol) was weighed and dissolved in CH ₂ Cl ₂ (40 mL), and the solution was placed in an ice bath, and DIPEA (4.6 g, 36 mmol) was slowly added dropwise with stirring. The mixed solution was slowly dropped into a zero-degree solution to which S-isopropyl(S)-2-aminopropylsulfate hydrochloride had been added, and the mixture was dropwise added in half an hour, and the solution was stirred at room temperature overnight.

The above solution was washed with water (300 mL), EtOAc (EtOAc) (EtOAc (EtOAc) g crude product.

Petroleum ether (100 mL), EtOAc (10 mL) was evaporated. The solution was filtered, washed with EtOAc (EtOAc) (EtOAc) (EtOAcjjjjjjj Propyl sulfate (1.07 g, 6%, epimer ratio is S _P /R _P =2.5:1).

^{1 H NMR (400MHz, MeOD)} δ7.2-6.4 (m, 4H), 4.2-3.8 (m, 1H), 3.5-3.3 (m, 1H), 1.9-1.6 (m, 1H), 1.4-1.2 ( m, 6H), 0.9-0.4 (m, 4H).

The second step S-isopropyl (S)-2-(((S)-(4-cyclopropylphenoxy)(((2R,3R,4R,5R)-5-(2,4-di) Preparation of Carbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propanesulfate

Take 1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H) - dione (0.60g, 2.4mmol) was dissolved in THF (20mL), cooled to 0 deg.] C, under protection of N ₂ was slowly added dropwise ^{t BuMgCl (1M in THF, 4.8mL} , 4.8mmol), after the addition was complete Stir at this temperature for 30 minutes, then warm to room temperature and continue stirring for 30 minutes. The reaction system was placed in an ice water bath, and S-isopropyl (2S)-2-((4-cyclopropylphenoxy)(perfluorophenoxy)phosphoryl)amino)propyl sulfate was slowly added dropwise. (1.0g, 1.9mmol) in THF (10mL) solution, the reaction was stirred overnight at this temperature, washed with 1N HCl solution (4mL) was quenched reaction was concentrated the residue was added CH _₂ Cl ₂ (50mL) and a 1N HCl solution (10 mL) The organic phase was separated and the aqueous phase was extracted with CH ₂ Cl ₂ (10 mL×2). The organic phase was combined and washed with saturated sodium hydrogen carbonate (10 mL) and brine (10 mL) Chromatography (eluent: CH ₂ Cl ₂ : ⁱ PrOH = 50:1 to 12:1) afforded the title compound S-isopropyl (S)-2-(((S)-(4-cyclopropylbenzene) Oxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4 Methyltetrahydrofuran-2-yl)methoxy)phosphoryl)amino)propylsulfate (0.28 g, 25%).

^{1 H NMR (400MHz, MeOD)} δ7.60 (d, J = 8.4Hz, 1H), 7.16 (m, 2H), 7.08 (d, J = 8.4Hz, 2H), 6.14 (d, J = 19.6Hz, 1H), 5.62 (d, J = 8.4 Hz, 1H), 4.55 (m, 1H), 4.42 (m, 1H), 4.13 (m, 1H), 3.98 (m, 2H), 3.52 (m, 1H), 1.93 (m, 1H), 1.39-1.33 (m, 6H), 1.27 (m, 6H), 0.96 (m, 2H), 0.65 (m, 2H);

³¹ P NMR (162 MHz, MeOD) δ 3.68;

¹⁹ F NMR (376 MHz, MeOD) δ -16.62;

MS m/z (ESI): 586.2 [M+H] ⁺ .

Biological evaluation

1. Determination of HCV inhibitory activity of hepatitis C virus

The inhibitory activity of the compounds of the present invention against HCV replication was determined using the HCV Replicon Reporter Luciferase Assay. The experimental cell model is the Huh-7 cell line stably transfected with the HCV luciferase reporter gene; the stock solution of the test compound is prepared as 10 mM with dimethyl sulfoxide, and the medium is diluted with a medium to a series of gradient concentrations, which are generally diluted into 8 to 10 concentrations. The internal reference compound tested was Cyclosporine.

The procedure was as follows: Cells were grown on 96-well plates, and different concentrations of test compound and internal reference compound were added to the cultured cells 24 hours later. After 48 hours, the luciferase activity was measured using a microplate reader. Compound half maximal inhibitory concentration IC ₅₀ values may be a range of different concentrations of test compound to inhibit the luciferase activity calculated by numerical.

2. Compound toxicity test on cells

The toxicity of the compounds of the invention to cells is determined by the MTT cytotoxicity assay. The experimental cell model is the Huh-7 cell line stably transfected with the HCV luciferase reporter gene; the stock solution of the test compound is prepared as 10 mM with dimethyl sulfoxide, and the medium is diluted with a medium to a series of gradient concentrations, which are generally diluted into 8 to 10 concentrations. The internal reference compound tested was Cyclosporine.

The procedure was as follows: Cells were grown on 96-well plates, and different concentrations of test compound and internal reference compound were added to the cultured cells 24 hours later. After 48 hours, MTT was added to the cultured cells for 4 hours, and then the absorbance was measured with a microplate reader. The half-inhibitory concentration CC ₅₀ value of the compound can be calculated from the inhibition of cell activity by the test compound at a range of different concentrations.

3. The list of bioactivity detection data of the compounds of the present invention is as follows:

Table 1

Table 2

** wherein PSI-7851 (S _P /R _P ≈ 1:1) is prepared according to the reference J. Med. Chem. 2010, 53, 7202, and its optically pure form Sofosbuvir (optical pure S _P ) is in accordance with reference J Prepared by .Org. Chem. 2011, 76, 8311.

Table 2 by a known preparation twenty-four the difference in the test results of Example isomers, optically pure active isomer cell configuration of the S _p R _p is superior isomer configuration.

4. PK test data of a preferred embodiment of the present invention

HCV NS5B is an RNA polymerase responsible for HCV virus replication, and the substrate used for replication is nucleoside triphosphate. The compound of the present invention is a phosphoramidate ester nucleotide analog prodrug which is metabolized in hepatocytes to produce an active drug nucleoside triphosphate analog, thereby inhibiting the activity of NS5B and the replication of HCV virus. As a prodrug, the ability to produce uridine triphosphate analogues in vivo is directly related to the inhibitory activity against NS5B, so the inventors performed an in vitro PK assay and an in vivo PK assay to detect the active compound of the preferred embodiment of the present invention. The ability of uridine phosphate analogues, where:

1) The in vitro PK test model is: PK analysis of Huh-7 cells and human primary hepatocytes. The specific test methods are as follows:

The pharmacokinetic test of prodrugs in human hepatocytes to produce the active drug fluorouridine triphosphate was carried out using human liver cancer Huh-7 cells and human primary hepatocytes. The drug was added to the culture solution 24 hours after cell colonization to a final concentration of 50 μM. After 24 hours of drug treatment, the cells were harvested with trypsin, counted, centrifuged, washed once with D-PBS, centrifuged, and the cell pellet was snap frozen in liquid nitrogen and stored in liquid nitrogen or -80 °C.

The concentration of the intracellular active drug fluorouridine triphosphate was measured by liquid chromatography-tandem mass spectrometry (LC/MS/MS). 1 ml of ice-cold 60% methanol was added to the frozen cell pellet to blow the cells and then left at -20 ° C overnight. The supernatant was obtained by centrifugation (11,000 rpm, 10 minutes) on the next day. 100 μL of the supernatant was taken from each sample, 100 μL of water was added, vortexed for 2 minutes, and centrifuged (3500 rpm, 5 minutes), and 10 μL of the supernatant was taken. The concentration of uridine triphosphate was measured by LC/MS/MS.

The instrument used for LC/MS/MS analysis of uridine triphosphate was AB Sciex API 400. The conditions of the liquid chromatography were Shimadzu LC-20AD pump, Ultimate X8-C8 column 4.6*250 mm, 5 μm column, the mobile phase used (A) solution was 10 mM ammonium acetate (pH = 7.8), and the (B) solution was acetonitrile at a flow rate of 0.56 mL/min, elution time 0-12.1 minutes, 95% (A) and 5% (B) for 0.01 minutes, 100% (A) for 4.2 minutes, 20% (A) and 80% for 4.5 minutes (B) ), 20% (A) and 80% (B) for 6 minutes, 95% (A) and 5% (B) for 6.1 minutes, 95% (A) and 5% (B) for 12 minutes, terminated by 12.1 minutes . The mass spectrometer was set up in the negative ion electrospray ionization (ESI) mode, and the multi-reaction monitoring of the analyte fluorouridine triphosphate was 499.2/158.7. The pharmacokinetic parameters were calculated using WinNonlin 6.1.

Preferred compounds of the present invention are the twenty-four, twenty-eight, thirty-two compounds of the examples. The main parameters of PK in vitro and comparison with Sofosbuvir are shown in Table 3:

Table 3. Main parameters of the in vitro PK assay of the compound of the example and comparison with Sofosbuvir

As can be seen from the data in Table 3, the compounds of the preferred embodiments of the present invention are the same as the Sofosbuvir, and the two in vitro PK test models can effectively metabolize the active drug uridine triphosphate. analog.

To further demonstrate the ability of the preferred embodiment compounds of the invention to metabolize the active drug triphosphate uridine analog, the inventors have also performed an in vivo PK assay to test the ability of the preferred embodiment compounds of the invention to produce the active drug uridine analog. The in vivo PK test model is: 1) liver PK analysis in rats; 2) liver PK analysis in dogs. The specific test methods are as follows:

1) PK analysis of rat liver

The pharmacokinetic test of the prodrug in the rat liver to produce the active drug fluorouridine triphosphate was carried out using SD rats (Shanghai Shrek). The administration is a single intragastric administration at a dose of 50 mg/10 ml/kg. The formulation of Sofosbuvir is 20% PEG200 and 0.5% sodium carboxymethylcellulose; the formulation of the twenty-four, twenty-eight, thirty-two compounds of the examples is 30% PEG200 and 0.5% sodium carboxymethylcellulose. The sampling points of the liver samples were 0.5, 1, 2, 4, 6, and 12 hours after administration. At the time of sampling, the rats were first sacrificed with CO ₂ , and the liver was washed with ice-cold saline through the portal vein, and the inferior vena cava was cut open for washing. 2/3 of the distal end of the left lobe of the liver was cut into approximately 0.2 gram pieces with a blade, snap frozen in liquid nitrogen, and stored in liquid nitrogen or -80 °C.

The active drug in the liver, fluorouridine triphosphate, is first separated from the uridine triphosphate by solid phase extraction, and then treated with alkaline phosphatase to remove the triphosphate to obtain uridine. The concentration of uridine is determined by liquid chromatography-tandem mass spectrometry. (LC/MS/MS) method was measured. Liver samples were spiked with 5 volumes of 60% methanol and homogenized; standards and controls were prepared with blank liver. Azidothymidine triphosphate (AZT-TP) was added to the liver homogenate as an internal reference. Solid phase extraction was performed using a weak anion exchange column from Waters Corporation, activated in 1 ml of methanol, 1 ml of water, 1 ml of potassium chloride containing 1% formic acid, and 1 ml of 1% formic acid. After homogenization of the liver, vortex for 2 minutes and then centrifuge at 4000 rpm for 10 minutes. The supernatant after centrifugation was applied to the activated column, and the column was washed with 1 ml of KCl containing 1% formic acid, and the nucleoside triphosphate was eluted twice with 0.3 ml of methanol containing 5% ammonia water. After drying with nitrogen at 37 ° C, it was redissolved with 160 μL of 5 mM ammonium acetate, and then treated with 40 μL of alkaline phosphatase at 37 ° C for 30 minutes to obtain a nucleoside. After the treatment, 20 μL was sampled for LC/MS/MS analysis of the concentration of the nucleoside.

The instrument used for LC/MS/MS analysis of nucleosides was AB Sciex API 5500Qtrap. The conditions of the liquid chromatography were Shimadzu LC-20AD pump, Luna 5 μm C18 30 x 2.0 mm column, the mobile phase used (A) solution was 5 mM ammonium acetate, and the (B) solution was 5 mM ammonium acetate / 95% acetonitrile / 5% water. The flow rate was 0.5 mL/min, the elution time was 0-4.5 minutes, the 0.01 minute was 100% (A), the 0.5 minute was 100% (A), and the 1.2 minute was 5% (A) and 95% (B), 2.4 minutes. 5% (A) and 95% (B), 100% (A) for 2.5 minutes, 100% (A) for 4.5 minutes. The setup conditions for the mass spectrometer are: negative ion electrospray ionization (ESI) mode, analyte The multi-reaction monitoring of fluorouridine was 259.20/239.20, and the multi-reaction monitoring of internal reference AZT was 266.10/223.10. The main parameters of pharmacokinetics were calculated using WinNonlin 6.1, and the measured data of the analyte fluorouridine was normalized according to the measurement data of the internal reference AZT.

2) PK analysis of canine liver

The pharmacokinetic test for the prodrug metabolism of the active drug in the whole liver to produce the active drug fluorouridine triphosphate was carried out with Beagle dogs (Beijing Musée). The administration was a single intragastric administration per day at a dose of 50 mg/10 ml/kg for 4 consecutive days. The formulations of Sofosbuvir and the twenty-four, twenty-eight, and thirty-two compounds of the examples were all 20% hydroxypropyl-β-cyclodextrin. The sampling point of the liver sample was 4 hours after the last administration. At the time of sampling, the dogs were first anesthetized with phenobarbital. The leaves of the liver were removed, washed briefly with ice-cold saline, and cut into small pieces of about 0.2 g with a blade, frozen in liquid nitrogen, and stored in liquid nitrogen or -80 ° C.

The active drug in the liver, fluorouridine triphosphate, is first separated from the uridine triphosphate by solid phase extraction, and then treated with alkaline phosphatase to remove the triphosphate to obtain uridine. The concentration of uridine is determined by liquid chromatography-tandem mass spectrometry. (LC/MS/MS) method was measured. Liver samples were spiked with 5 volumes of 60% methanol and homogenized; standards and controls were prepared with blank liver. Azidothymidine triphosphate (AZT-TP) was added to the liver homogenate as an internal reference. Solid phase extraction was performed using a weak anion exchange column from Waters Corporation, activated in 1 ml of methanol, 1 ml of water, 1 ml of potassium chloride containing 1% formic acid, and 1 ml of 1% formic acid. After homogenization of the liver, vortex for 2 minutes and then centrifuge at 4000 rpm for 10 minutes. The supernatant after centrifugation was applied to the activated column, and the column was washed with 1 ml of KCl containing 1% formic acid, and the nucleoside triphosphate was eluted twice with 0.3 ml of methanol containing 5% ammonia water, and the eluate was at 37. After drying at room temperature with nitrogen, the solution was re-dissolved with 160 μL of 5 mM ammonium acetate, and then treated with 40 μL of alkaline phosphatase at 37 ° C for 30 minutes to obtain a nucleoside. After the treatment, 20 μL was sampled for LC/MS/MS analysis of the concentration of the nucleoside.

The instrument used for LC/MS/MS analysis of nucleosides was AB Sciex API 5500Qtrap. The conditions of the liquid chromatography were Shimadzu LC-20AD pump, Luna 5 μm C18 30 x 2.0 mm column, the mobile phase used (A) solution was 5 mM ammonium acetate, and the (B) solution was 5 mM ammonium acetate / 95% acetonitrile / 5% water. The flow rate was 0.5 mL/min, the elution time was 0-4.5 minutes, the 0.01 minute was 100% (A), the 0.5 minute was 100% (A), and the 1.2 minute was 5% (A) and 95% (B), 2.4 minutes. 5% (A) and 95% (B), 100% (A) for 2.5 minutes, 100% (A) for 4.5 minutes. The setup conditions for the mass spectrometer are: negative ion electrospray ionization (ESI) mode, analyte Fluorine The multi-reaction monitoring of uridine was 259.20/239.20, and the multi-reaction monitoring of internal reference AZT was 266.10/223.10. The main parameters of pharmacokinetics were calculated using WinNonlin 6.1, and the measured data of the analyte fluorouridine was normalized according to the measurement data of the internal reference AZT.

Preferred compounds of the present invention are the twenty-four, twenty-eight, thirty-two compounds of the examples. The main parameters of PK in vivo and comparison with Sofosbuvir are shown in Table 4:

Table 4. Main parameters of the PK test in vivo of the example compounds and comparison with Sofosbuvir

As can be seen from the data in Table 4, the compounds of the preferred embodiments of the present invention, the twenty-four, twenty-eight, thirty-two compounds, like Sofosbuvir, are also effective in metabolizing the active drug uridine uridine analogs in vivo, especially It is the ability of the compound of the thirty-two compound to produce uridine triphosphate analog in rat liver and canine liver compared with the positive drug Sofosbuvir, the data are improved by more than 20%, and the twenty-fourth compound in the canine liver The ability to produce uridine triphosphate analogs was increased by nearly 35% compared to the positive drug Sofosbuvir. Compared with the in vitro hepatocyte PK experiment, the in vivo experiment integrates the processes of drug absorption, protein binding, tissue distribution and metabolism, and more reflects the true condition of the compound, and therefore has more clinical significance.

It should be noted that the above embodiments are only intended to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art The technical means is modified or equivalently substituted. For example, the first to the twenty-third examples can be separated by column chromatography to obtain the (S)-isomer or the (S)-isomer can be synthesized according to the method of the twenty-fourth embodiment. The technical solutions are also intended to be within the scope of the claims of the invention.

Claims

A uracil nucleotide analog of the compound of formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof:

Wherein Z is selected from oxygen or sulfur; Y is selected from hydrogen or acetyl;

R 1 is selected from the group consisting of hydrogen, halogen, hydroxy, decyl, cyano, nitro, azide, C 1-8 alkyl, halo-substituted C 1-8 alkyl, C 2-8 alkenyl, C 2-8 Alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C 5-10 aryl, C 5 10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C(O)OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 C(O)NR 6 R 7 , -N(R 5 )-C(O)R 5 , -N(R 5 )-C(O)OR 5 , -C 0-8 - SR 5 , -SiR 5 R 6 R 7 , -GeR 5 R 6 R 7 ,

or

The carbon atom adjacent to the benzene ring of R 1 forms a 5-7 membered carbon ring and a 5-7 membered heterocyclic ring.

Wherein C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic, C 5-10 aryl, 5-10 membered heteroaryl, 5-7 membered carbocyclic or 5-7 membered heterocyclic ring optionally further substituted with one or more substituents selected from halo, hydroxy, mercapto, cyano, nitro, azido, C 1- 8- alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclic oxy, 3-8 Heterocyclylthio, C 5-10 aryl, C 5-10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5 -10-membered heteroarylthio, -C 0-8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 - C(O)OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 -C(O)NR 6 R 7 , -N(R 5) -C (O) R 5 or -N (R 5) -C (O ) oR 5 is substituted with a substituent;

R 2 is selected from the group consisting of hydrogen, halogen, hydroxy, decyl, cyano, nitro, azide, C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 ring Alkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C 5-10 aryl, C 5-10 aryloxy, C 5 10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C 0-8 -S(O)rR 5 , -C 0 -8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C(O)OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 -C(O)NR 6 R 7 , -N(R 5 )-C(O)R 5 or -N(R 5 )-C(O)OR 5 Substituted by

Wherein the C 1-8 alkyl group, the C 3-8 cycloalkyl group, the 3-8 membered heterocyclic group, the C 5-10 aryl group or the 5-10 membered heteroaryl group is optionally further selected by one or more From halogen, hydroxy, thiol, cyano, nitro, azide, C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl, 3- 8-membered heterocyclic group, 3-8 membered heterocyclic oxy group, 3-8 membered heterocyclic thio group, C 5-10 aryl group, C 5-10 aryloxy group, C 5-10 arylthio group 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C 0-8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C(O)OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 Substituted with a substituent of -C 0-8 -C(O)NR 6 R 7 , -N(R 5 )-C(O)R 5 or -N(R 5 )-C(O)OR 5 ;

R 3 is selected from hydrogen, C 1-8 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 5-10 aryl or 5-10 membered heteroaryl, optionally further by one or a plurality selected from the group consisting of halogen, hydroxy, decyl, cyano, nitro, azide, C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl , 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic thio group, C 5-10 aryl, C 5-10 aryloxy, C 5-10 aryl Thiothio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C 0-8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C(O)OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR a substituent of 6 R 7 , -C 0-8 -C(O)NR 6 R 7 , -N(R 5 )-C(O)R 5 or -N(R 5 )-C(O)OR 5 Replace

R 4 is selected from the group consisting of hydrogen, halogen, hydroxy, decyl, cyano, nitro, azide, C 1-8 alkyl, halo-substituted C 1-8 alkyl, C 3-8 cycloalkyl, C 3-8 Cycloalkylmethyl, halo-substituted C 1-8 alkoxy, halo-substituted C 1-8 alkylthio, 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic ring Thiothio, C 5-10 aryl, C 5-10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 N-heteroarylthio, -C 0-8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C( O) OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 -C(O)NR 6 R 7 , -N(R 5 ) -C(O)R 5 , -N(R 5 )-C(O)OR 5 ;

R 5 , R 6 , and R 7 are selected from the group consisting of hydrogen, C 4 -alkyl, C 3-8 cycloalkyl;

m is 0, 1, 2, 3, 4;

r is 0, 1, 2.
The uracil nucleotide analog according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein Z is selected from the group consisting of oxygen, and the formula is a compound of the formula (II).

Wherein: Y, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 2, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein

R 1 is selected from C 1-8 alkyl, halo substituted C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic ring , 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C 5-10 aryl, C 5-10 aryloxy, C 5-10 arylthio, 5-10 a heteroaryl group, a 5-10 membered heteroaryloxy group, a 5-10 membered heteroarylthio group, -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0 -8 -C(O)OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 -C(O)NR 6 R 7 ,- N(R 5 )-C(O)R 5 , -N(R 5 )-C(O)OR 5 , -C 0-8 -SR 5 , -SiR 5 R 6 R 7 , -GeR 5 R 6 R 7 ,

Wherein C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 5-10 aryl or The 5-10 membered heteroaryl group is further optionally further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide, C 1-8 alkyl, C 2-8 alkenyl, C 2 -8 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C 5-10 aryl, C 5-10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C 0-8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C(O)OR 5 , -C 0-8 -OC (O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 C(O)NR 6 R 7 , -N(R 5 )-C(O)R 5 or -N(R 5 Substituted by a substituent of -C(O)OR 5 ;

Y, R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 3, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein

R 1 is selected from the group consisting of C 1-8 alkyl, halo-substituted C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl,

Wherein the C 1-8 alkyl group, the halogen-substituted C 1-8 alkyl group, the C 2-8 alkenyl group or the C 2-8 alkynyl group is further one or more selected from the group consisting of C 3-8 cycloalkyl groups , 3-8 membered heterocyclyl, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic thio group, C 5-10 aryl, C 5-10 aryloxy, C 5-10 aryl Substituted with a substituent of a thiol group, a 5-10 membered heteroaryl group, a 5-10 membered heteroaryloxy group or a 5-10 membered heteroarylthio group;

Y, R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 4, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of the following compounds:
The uracil nucleotide analog according to claim 5, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of the following compounds:
The uracil nucleotide analog according to claim 3, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein

R 1 is selected from C 3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C 5-10 aryl, C 5 - 10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio,

Wherein the C 3-8 cycloalkyl group, the 3-8 membered heterocyclic group, the C 5-10 aryl group or the 5-10 membered heteroaryl group is further further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, Cyano, nitro, azide, C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic, 3 -8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C 5-10 aryl, C 5-10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl , 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C 0-8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 - C(O)R 5 , -C 0-8 -C(O)OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 - Substituted by a substituent of C(O)NR 6 R 7 , -N(R 5 )-C(O)R 5 or -N(R 5 )-C(O)OR 5 ;

Y, R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 7, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of the following compounds:
The uracil nucleotide analog according to claim 8, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of the following compounds:
The uracil nucleotide analog according to claim 3, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from the group consisting of -C 0-8 -SR 5 and -SiR 5 R 6 R 7 , -GeR 5 R 6 R 7 ,

Y, R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 10, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of the following compounds:
The uracil nucleotide analog according to claim 11, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of the following compounds:
The uracil nucleotide analogues according to claim 2, which is a stereoisomer or a pharmaceutically acceptable salt thereof, wherein, R 1 and the phenyl ring adjacent carbon atoms form a 5-7 membered carbocyclic ring, 5-7 yuan heterocyclic ring,

Wherein the 5-7 membered carbocyclic or 5-7 membered heterocyclic ring is optionally further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide, C1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic thio , C 5-10 aryl, C 5-10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered Arylthio, -C 0-8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C(O) OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 -C(O)NR 6 R 7 , -N(R 5 )-C Substituting (O) a substituent of R 5 or -N(R 5 )-C(O)OR 5 ;

Y, R 3 , R 4 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 13, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R 1 and a carbon atom adjacent to the benzene ring form a 5-7 membered carbocyclic ring, The 5-7 membered heterocyclic ring is selected from the following structures:

Wherein the 5-7 membered carbocyclic or 5-7 membered heterocyclic ring is optionally further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide, C1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic thio , C 5-10 aryl, C 5-10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered Arylthio, -C 0-8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C(O) OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 -C(O)NR 6 R 7 , -N(R 5 )-C Substituting (O) a substituent of R 5 or -N(R 5 )-C(O)OR 5 ;

Y, R 3 , R 4 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analogues according to claim 14, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 1 and the phenyl ring adjacent carbon atoms form a 5-7 membered carbocyclic ring, The 5-7 membered heterocyclic ring is selected from the following structures:
The uracil nucleotide analog according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein Z is selected from the group consisting of sulfur, and the formula is a compound of the formula (III).

Wherein Y, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , m, and r are as defined in claim 1.
The uracil nucleotide analog according to claim 16, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein

R 4 is selected from the group consisting of halogen, hydroxy, decyl, C 1-8 alkyl, halo-substituted C 1-8 alkyl, C 3-8 cycloalkyl, C 3-8 cycloalkylmethyl, halo-substituted C 1-8 alkane An oxy group, a halogen-substituted C 1-8 alkylthio group;

Y, R 1 , R 2 , R 3 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 17, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R 3 is selected from the group consisting of hydrogen, C 1-8 alkyl, and C 3-8 ring An alkyl group, a 3-8 membered heterocyclic group, a C 5-10 aryl group or a 5-10 membered heteroaryl group, optionally further selected from one or more selected from the group consisting of halogen, hydroxy, decyl, C 1-8 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic group, 3-8 membered heterocyclic oxy group, 3-8 membered heterocyclic thio group, -C 0-8 -S(O)rR 5 , -C Substituted by a substituent of 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C(O)OR 5 or -C 0-8 -OC(O)R 5 ; R 4 is selected from the group consisting of fluorine, methyl, trifluoromethyl, cyclopropyl, cyclopropylmethyl;

Y, R 1 , R 2 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 18, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R 3 is selected from the group consisting of hydrogen, C 1-4 alkyl, cyclopropyl, and ring. Hexyl or phenyl, optionally further selected from one or more selected from the group consisting of halogen, hydroxy, decyl, -C 0-8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 -C Substituting (O) R 5 , -C 0-8 -C(O)OR 5 or -C 0-8 -OC(O)R 5 ; R 4 is selected from methyl;

Y, R 1 , R 2 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 19, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from the group consisting of hydrogen, C 1-8 alkyl, and halogen substituted C 1- 8- alkyl, C 2-8 alkenyl, C 2-8 alkynyl, wherein said C 1-8 alkyl, halo substituted C 1-8 alkyl, C 2-8 alkenyl or C 2 The -8 alkynyl group is further one or more selected from the group consisting of C 3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C 5-10 aryl, C 5-10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy or 5-10 membered heteroarylsulfide Substituted by a substituent of the group;

Y, R 2 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 20, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of the following compounds:
The uracil nucleotide analog according to claim 19, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from the group consisting of C 3-8 cycloalkyl and 3-8 membered heterocyclic ring. , 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C 5-10 aryl, C 5-10 aryloxy, C 5-10 arylthio, 5-10 a heteroaryl group, a 5-10 membered heteroaryloxy group, a 5-10 membered heteroarylthio group, wherein the C 3-8 cycloalkyl group, a 3-8 membered heterocyclic group, a C 5-10 aryl group Or a 5-10 membered heteroaryl optionally further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide, C1-8 alkyl, C2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C 5-10 aryl , C 5-10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered heteroarylthio, -C 0 -8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C(O)OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 -C(O)NR 6 R 7 , -N(R 5 )-C(O)R 5 or -N Substituted by a substituent of (R 5 )-C(O)OR 5 ;

Y, R 2 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 22, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of the following compounds:
The uracil nucleotide analog according to claim 19, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from the group consisting of -C 0-8 -SR 5 and -SiR 5 R 6 R 7 , -GeR 5 R 6 R 7 ;

Y, R 2 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 24, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of the following compounds:
The uracil nucleotide analog according to claim 19, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein

The carbon atom adjacent to the benzene ring of R 1 forms a 5-7 membered carbon ring and a 5-7 membered heterocyclic ring.

Wherein the 5-7 membered carbocyclic or 5-7 membered heterocyclic ring is optionally further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide, C1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic thio , C 5-10 aryl, C 5-10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered Arylthio, -C 0-8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C(O) OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 -C(O)NR 6 R 7 , -N(R 5 )-C Substituting (O) a substituent of R 5 or -N(R 5 )-C(O)OR 5 ;

Y, R 2 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
The uracil nucleotide analog according to claim 26, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R 1 and a carbon atom adjacent to the benzene ring form a 5-7 membered carbocyclic ring, The 5-7 membered heterocyclic ring is selected from the following structures:

Wherein the 5-7 membered carbocyclic or 5-7 membered heterocyclic ring is optionally further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide, C1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclic thio , C 5-10 aryl, C 5-10 aryloxy, C 5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 membered Arylthio, -C 0-8 -S(O)rR 5 , -C 0-8 -OR 5 , -C 0-8 -C(O)R 5 , -C 0-8 -C(O) OR 5 , -C 0-8 -OC(O)R 5 , -C 0-8 -NR 6 R 7 , -C 0-8 -C(O)NR 6 R 7 , -N(R 5 )-C Substituting (O) a substituent of R 5 or -N(R 5 )-C(O)OR 5 ;

Y, R 2 , R 5 , R 6 , R 7 , m, r are as defined for the compound of formula (I).
The uracil nucleotide analog according to claim 27, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of the following compounds:
The uracil nucleotide analog according to any one of claims 16 to 28, which is a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the stereoisomer is in the S configuration and has the following structure: :
The uracil nucleotide analog according to claim 29, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of the following compounds:
A method for preparing a uracil nucleotide analog according to claim 1, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, comprising the steps of:

When Y is selected from the group consisting of acetyl groups, the optional further includes the following reactions:

Optionally, further comprising column chromatography to obtain the stereoisomer thereof, or obtaining the stereoisomer thereof by the following steps:

When Y is selected from the group consisting of acetyl groups, the optional further includes the following reactions:

Wherein: Y, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , m, r are as defined in claim 1.
A pharmaceutical composition comprising a therapeutically effective amount of a uracil nucleotide analog according to claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
The uracil nucleotide analog according to claim 1, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 32, which is prepared for the treatment of hepatitis C virus, type A Use of drugs for diseases caused by hepatitis virus, West Nile virus, yellow fever virus, dengue virus, rhinovirus, poliovirus, bovine viral diarrhea virus or Japanese encephalitis virus infection.