TW201605885A - Uracil nucleotide analogues, their preparation method and use thereof - Google Patents

Abstract

The invention relates to uracil nucleotide analogues, their preparation method and use thereof. Specifically, the invention provides the uracil nucleotide analogues with the formula (I), their stereoisomers and pharmaceutical acceptable salts, the preparation method and use. These compounds are RNA-dependent RNA viral replication inhibitors, and can be used as HCV NS5B polymerase inhibitors, HCV replication inhibitors as well as for the treatment of hepatitis C infection in mammals. They have broad application prospects and are expected to develop a new generation of antiviral drugs.

Description

Uracil nucleotide analog and preparation method and application thereof

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.

The virus of the Flaviviridae family includes at least three different genera: Pestiviruses, which cause disease in cattle and pigs; Flavivruses, which are the main cause of diseases such as dengue and yellow fever; 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, and in 1989, American scientist Michael. Used by Michael Houghton and his colleagues The molecular biology of the virus was found in the molecular biology method, and the hepatitis C virus was cloned. The disease and its virus were named 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 length of approximately 3014 amino acids A proprotein 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 compare 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 patients with chronic hepatitis C 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 use of imported clotting 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 serious liver disease tendency: 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 From 0% to 20% of chronically infected individuals eventually develop liver-destructive sclerosis or cancer. The viral disease is transmitted parenterally by contaminated blood and blood products, contaminated needles; or by sexual transmission; and from the mother of the infected mother or carrier to the offspring of the carrier. 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 4OKD 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 The whole body 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, so the recurrence rate after stopping the drug is low. 4OKD macromolecular pegylated interferon is limited to vascular and intrahepatic distribution due to excessive 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-natural matrix 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 usually mediated by a cellular kinase that targets the potential nucleus Inosin polymerase inhibitors present additional structural requirements. 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 hampered by poor matrix properties relative to one or more kinases that are required to convert the nucleoside to the active triphosphate form. The formation of a monophosphate by nucleoside kinase is generally considered to be a rate determining step in the process of triphosphorylation. 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 aminophosphate 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 confirmed that nucleoside aminophosphate prodrugs improve the systemic absorption of nucleosides, and further, the aminophosphate moieties of these "original nucleotides" are partially masked by neutral lipophilic groups to obtain suitable partition coefficients to optimize The uptake and entry 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 without the need for an initial monophosphorylation rate determining step.

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

The inventors discovered a class of uracil nucleotide analogs during the course of their research, which are inhibitors of RNA-dependent RNA viral replication and are useful as inhibitors of HCV NS5B polymerase, inhibitors of HCV replication, and Hepatitis C infection in mammals has broad application prospects 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 ethoxylated; R _{1 is} selected from the group consisting of hydrogen, halogen, hydroxy, thiol, cyano, nitro, azide, C _1-8 alkyl, halogen 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 a carbon atom adjacent to the benzene ring of R ₁ forms 5 a -7 membered carbocyclic ring, a 5-7 membered heterocyclic ring wherein the C _1-8 alkyl group, C _2-8 alkenyl group, C _2-8 alkynyl group, C _3-8 cycloalkyl group, 3-8 member a heterocyclic group, a C _5-10 aryl group, a 5-10 membered heteroaryl group, a 5-7 membered carbocyclic ring or a 5-7 membered heterocyclic ring, further optionally one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, Nitro, azido, 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; 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 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, 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 group is substituted with substituents; wherein the C _1-8 alkyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, C _5-10 aryl, 5-10 membered heteroaryl optionally further substituted by 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 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 for a substituent of ₅ )-C(O)OR ₅ ; R _{3 is} selected from the group consisting of hydrogen, C _1-8 alkyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, C _5-10 aryl Or a 5-10 membered heteroaryl group, further optionally 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 heteroaryl group 5-10 membered heteroaryl _{group, -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 ; R _{4 is} selected from the group consisting of hydrogen, halogen, hydroxy, thiol, cyano, nitro, azide , C _1-8 alkyl, halogen substituted C _1-8 alkyl, C _3-8 cycloalkyl, C _3-8 cycloalkylmethyl, halogen substituted C _1-8 alkoxy, halogen substituted C _{1- 8} -alkylthio, 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 , -N(R ₅ )-C(O)OR ₅ ; R ₅ , R ₆ , and R _{7 are} selected from the group consisting of hydrogen, C _1-4 alkyl, and C _3-8 cycloalkyl; and m is 0, ₁ , 2, 3, 4.

r is 0, 1, 2

As used herein, a "5-7 membered carbocyclic ring" refers to a percarbocyclic ring containing from 5 to 7 carbon atoms, including a cycloalkyl or aryl group, and a "5-7 membered heterocyclic ring" refers to 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 the group consisting of C _1-8 alkyl, halogen substituted 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, 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 the C _1-8 alkyl group, C _2-8 alkenyl group, C _{2 -8} alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, C _5-10 aryl or 5-10 membered heteroaryl optionally further selected from one or more selected from the group consisting of halogen, hydroxy, Mercapto, 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 heterocyclic oxy, 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) Substituted by a substituent of 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 the group consisting of C _1-8 alkyl, halogen substituted C _1-8 alkyl, C _{a 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 alkynyl group is further a plurality selected from C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclyloxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _{5 -} Substituted by a substituent of a ₁₀ aryloxy group, a C _5-10 arylthio group, a 5-10 membered heteroaryl group, a 5-10 membered heteroaryloxy group or a 5-10 membered heteroarylthio group; 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 pharmaceutically acceptable salt thereof is selected from the group consisting of the following compounds:

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 the group consisting of C _3-8 cycloalkyl, 3-8 membered heterocyclic group, 3 -8 membered heterocyclooxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 member heteroaryl 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 The heteroaryl group is 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 chain. 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 )- C (O) OR ₅ substituents being _{substituted; Y, R 2, R 3} , R 4, R 5, R 6 R _7, m, r compound of formula (I) as defined above.

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 the following compounds:

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 _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:

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, the carbon atom adjacent to the benzene ring of R ₁ forms a 5-7 membered carbocyclic ring, 5-7 Heterocyclic ring, wherein the 5-7 membered carbocyclic ring or the 5-7 membered heterocyclic ring is further 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 group Sulfur, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5-10 member 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 )- Substituted by a substituent of C(O)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 formula (I) ) defined by the compound.

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 heteroaryl surveying requires further by 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 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 heteroarylsulfide Base, -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) Substituted by 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) .

Preferably, the uracil nucleotide analog, its stereoisomer or a pharmaceutically acceptable salt thereof, the carbon atom adjacent to the benzene ring of R ₁ forms a 5-7 membered carbon ring, 5-7 member The 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, 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, 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 the group consisting of hydrogen, C _1-4 alkyl, cyclopropyl, cyclohexyl or Phenyl, further optionally 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 alkyne group Further 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 a substituent of a C _5-10 aryloxy group, a C _5-10 arylthio group, a 5-10 membered heteroaryl group, a 5-10 membered heteroaryloxy group or a 5-10 membered heteroarylthio group Substituted; Y, R ₂ , R ₅ , R ₆ , R ₇ , m, r are as defined in the first item of the patent application.

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 the following compounds:

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 heterocyclooxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 member An aryl 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- The 10-membered heteroaryl group is 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 ) -C (O) OR ₅ substituents being _{substituted; Y, R 2, R 5} , R 6, R 7, m R The compounds of formula (I) as defined above.

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 the following compounds:

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 -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 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 the following compounds:

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- a 7-membered heterocyclic ring, wherein the 5-7 membered carbocyclic or 5-7 membered heterocyclic ring is further further selected from one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, azide, _C1-8 alkane, as desired , C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 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 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 ) Substituted by a substituent of -C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ; Y, R ₂ , R ₅ , R ₆ , R ₇ , m, r such as a compound of formula (I) Defined.

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 heteroaryl surveying requires further by 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 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 heteroarylsulfide Base, -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) Substituents of R ₅ or -N(R ₅ )-C(O)OR ₅ are substituted; 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 the following compounds:

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

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 the following compounds:

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 ethyl hydrazino, the reaction further includes the following reaction as needed: Further, if necessary, column chromatography is carried out to obtain a stereoisomer thereof, or a stereoisomer thereof is obtained by the following steps: When Y is selected from the group consisting of ethyl hydrazino, the reaction further includes the following reaction as needed:

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).

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 analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the treatment of hepatitis C virus, A, using the compound of formula (I) Use of drugs for diseases caused by hepatitis B virus, West Nyrovirus, yellow fever virus, dengue virus, rhinovirus, poliovirus, bovine viral diarrhea virus or Japanese encephalitis virus infection.

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-dimethylpropane 1,1,2-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-glycol Base, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2- Dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethyl Hexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethyl Hexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl or various branched isomers thereof.

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, fluorenyl. , 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 heterocyclooxy, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 member Aryl, 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 saturation Or a partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, "C _3-8 cycloalkyl" refers to a cycloalkyl group comprising from 3 to 8 carbon atoms, and "5-10 membered cycloalkyl" refers to 5 to Non-limiting examples of a cycloalkyl group of 10 carbon atoms, for example, a monocyclic cycloalkyl group, include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclopentenyl group, and a ring. Group, cyclohexenyl group, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, 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. The spirocycloalkyl group is divided into a monospirocycloalkyl group, a bispirocycloalkyl group or a polyspirocycloalkyl group according to the number of shared spiro atoms between the ring and the ring. Non-limiting examples of spirocycloalkyl groups include:

"Fused cycloalkyl" means 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, tetrahydronaphthalene Base, 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 member heteroaryl Alkoxy group, 5-10 membered heteroarylthio group, -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 by a substituent of -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ; "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic ring a cyclic hydrocarbon substituent wherein one or more ring atoms are selected from nitrogen, oxygen or S(O)r (wherein r is an integer 0, 1, 2) heteroatom, but does not include -OO-, -OS- or - The ring portion of SS-, the remaining ring atoms are 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.

The polycyclic heterocyclic group includes a spiro ring, a fused ring, and a heterocyclic group of a bridged ring. "Spiroheterocyclyl" refers to a polycyclic heterocyclic group in which one atom (called a spiro atom) is shared between monocyclic rings, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O)r (where r is an integer) The heteroatoms of 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, and Nitrogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclooxy Base, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 member Aryloxy, 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; "aryl" refers to an all carbon monocyclic or fused polycyclic ( That is, a ring group sharing a pair of adjacent carbon atoms, a polycyclic ring having a conjugated π-electron system (ie, a ring having an adjacent pair of carbon atoms), and a "C _5-10 aryl group" containing 5 - 10 carbons of all-carbon aryl, "5-10 membered aryl" means an all-carbon aryl group containing 5-10 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 may be 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 heteroaryloxy Base, 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 by a substituent of N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ; "heteroaryl" means a heteroaromatic system containing from 1 to 4 heteroatoms The hetero atom includes a hetero atom of nitrogen, oxygen and S(O)r (wherein r is an integer of 0, 1, 2), and a 5-7 membered heteroaryl group means a heteroaromatic system having 5 to 7 ring atoms. 5-10 membered heteroaryl refers to a heteroaromatic system containing 5-10 ring atoms, such as furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, Oxazolyl, tetrazolyl and the like. The heteroaryl ring can 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 member heteroaryl Alkoxy group, 5-10 membered heteroarylthio group, -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 by a substituent of -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ; "alkenyl" means at least two carbon atoms and at least one carbon - An alkyl group as defined above having a carbon double bond, and a C _2-8 alkenyl group means a straight or branched chain alkenyl group having 2-8 carbons. For example, vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, and the like.

The alkenyl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from 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 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 Substituted by a substituent of (R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ; "alkynyl" means at least two carbon atoms and at least one carbon-carbon triple bond The alkyl group as defined above, C _2-8 alkynyl refers to a straight or branched alkynyl group having 2-8 carbons. For example, ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, and the like.

The alkynyl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from 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 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 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 defined As mentioned 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 is preferably one or more of the following groups independently selected from the group consisting of halogen, hydroxy, decyl, cyano, nitro, and Nitrogen, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclooxy Base, 3-8 membered heterocyclylthio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 member Aryloxy, 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; "cycloalkoxy" and 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 member heteroaryl Alkoxy group, 5-10 membered heteroarylthio group, -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 by a substituent of -N(R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ; "halogen-substituted C _1-8 alkyl" refers to an alkyl group Hydrogen 1-8 alkylalkyl groups substituted by fluorine, chlorine, bromine or iodine atoms, such as difluoromethyl, dichloromethyl, dibromomethyl, trifluoromethyl, trichloromethyl , tribromomethyl and the like.

The hydrogen on the "halogen-substituted C _1-8 alkoxy" alkyl group is preferably 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.

"As needed" or "as needed" 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 a case where a heterocyclic group is substituted with an alkyl group and a case where a 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 amine 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 shaping Agent. The purpose of the pharmaceutical composition is to promote the administration of the organism, and to facilitate the absorption of the active ingredient to exert 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 hydrazine (DMSO- d ₆ ). The deuterated methanol (CD ₃ OD) and deuterated chloroform (CDCl ₃ ) were internally labeled as four Base decane (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 chromatography column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18 150 x 4.6 mm chromatography column).

The thin layer chromatography tantalum sheet uses Yantai Yellow Sea HSGF254 or Qingdao GF254 tantalum sheet. The specification for TLC is 0.15mm to 0.20mm, and the specification for thin layer chromatography separation and purification is 0.4mm to 0.5mm. Pipe column chromatography generally uses Yantai Huanghai Tanji 200~300 mesh silicone as carrier.

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

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

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. The system of the eluent for column chromatography includes: A: dichloromethane and methanol systems, B: n-hexane and ethyl acetate systems, C: dichloromethane and ethyl acetate systems, D: ethyl acetate and methanol, The volume ratio of the solvent is adjusted depending on the polarity of the compound, and may be adjusted by adding a small amount of ammonia water, acetic acid or the like.

Embodiment 1

Preparation of the first step ((2-cyclopropylphenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester

4-Nitrophenyl dichlorophosphate (320 mg, 1.25 mmol) was dissolved in CH ₂ Cl ₂ (2.5 mL), cooled to -78 ° C, 2-cyclopropyl phenol (185 mg, 1.38 mmol) and TEA (192 μ L, 1.38 mmol) of CH ₂ Cl ₂ (2.5 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- at 0 ° C. alanine isopropyl ester hydrochloride (210mg, 1.25mmol) in CH ₂ Cl ₂ (2.5mL) solution, followed by TEA (366 μ L, 2.63mmol) was added dropwise to the reaction system was stirred at 0 ℃ 1 hour The reaction mixture was concentrated under reduced EtOAc. Column chromatography (eluent: PE: EtOAc = 5:1) gave the title compound ((2-cyclopropylphenoxy)(4-nitrophenoxy)phosphonyl)-L-alanine Propyl ester (395 mg, 70%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.16-8.23 (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).

Second step ((2-cyclopropylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl) Preparation of -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl ester

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 (1M, 0.45 mL, 0.45 mmol) was added dropwise to the above solution at room temperature. Under stirring for 10 minutes, ((2-cyclopropylphenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester (200 mg, 0.45) was added dropwise to the reaction. A solution of mmol in THF (1.5 mL) was stirred at 55 ° C overnight. Then cooled to room temperature, methanol (1 mL) the reaction was quenched, concentrated to column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 30: 1) under reduced pressure to give the title compound ((2-cyclopropyl Phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- 4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl ester (15 mg, 12%, epimer ratio: S _P /R _P =4.1:1) .

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.80 (s, 1H), 7.44 (d, J = 8.0Hz, 1H), 7.35 (d, J = 8.0Hz, 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 (162MHz, CDCl ₃ ): δ 4.40, 3.50; MS m/z (ESI): 570.1 [M+H] ⁺ .

Embodiment 2

Preparation of the first step ((3-cyclopropylphenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester

Dichloro-4-nitrophenyl phosphate (1.950g, 7.62mmol) was dissolved in CH ₂ Cl ₂ (15mL), cooled to -78 ℃, 3- cyclopropyl-phenol (1.124g, 8.38mmol) and TEA ( A solution of 1.17 mL, 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, and the reaction solution was added dropwise to the cooled L-propylamine 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 ((3-cyclopropylphenoxy)(4-nitrophenoxy)phosphonyl)-L-alanine Propyl ester (2.905 g, 85%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.17-8.26 (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 ((3-cyclopropylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl) Preparation of -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl ester

1-((2R,3R,4R,5S)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)- The 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. Under stirring for 10 minutes, ((3-cyclopropylphenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester (938 mg, 2.09) was added dropwise to the reaction. A solution of mmol in 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 ((3-cyclopropyl Phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- 4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl ester (20 mg, 3%, epimer ratio: S _P /R _P =3.8:1) .

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.74 (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

Preparation of the first step ((2-cyclopropyl-6-methylphenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester

4-Nitrophenyl dichlorophosphate (1.950 g, 7.62 mmol) was dissolved in CH ₂ Cl ₂ (15 mL), cooled to -78 ° C, 2-cyclopropyl-6-methylphenol (1.129 g, 7.62 Ment) and TEA (1.17 mL, 8.39 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 to 0 ° C. the cooled 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) gave the title compound ((2-cyclopropyl-6-methylphenoxy)(4-nitrophenoxy) Mercapto)-L-isopropyl isopropyl ester (2.880 g, 82%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.18 (d, J = 9.2Hz, 2H), 7.32 (dd, J = 8.4,7.2Hz, 2H), 7.00 (d, J = 4.8Hz, 2H), 6.68 -6.75 (m, 1H), 4.91-5.05 (m, 1H), 4.05-4.21 (m, 1H), 3.89-4.05 (m, 1H), 2.39 (s, 3H), 2.17-2.31 (m, 1H) , 1.13-1.42 (m, 12H), 0.90-1.06 (m, 2H), 0.59-0.75 (m, 2H).

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

1-((2R,3R,4R,5S)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)- The 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. Under stirring for 10 minutes, ((2-cyclopropyl-6-methylphenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl was added dropwise to the reaction. A solution of the ester (967 mg, 2.09 mmol) in THF (6 mL) 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 ((2-cyclopropyl -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)phosphonium)-L-alanine isopropyl ester (20 mg, 3%, epimer ratio S _P /R _P >10:1).

_{^{1 H NMR (400MHz, CD 3}} OD): δ 7.53 (d, J = 8.0Hz, 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

Preparation of the first step ((4-cyclopropylphenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester

4-Nitrophenyl dichlorophosphate (1.260 g, 4.92 mmol) was dissolved in CH ₂ Cl ₂ (10 mL), cooled to -78 ° C, 4-cyclopropyl phenol (726 mg, 5.41 mmol) and TEA (0.76) A solution of mL, 5.45 mmol) in CH ₂ Cl ₂ (10 mL) was added dropwise at this temperature for 30 minutes, then gradually warmed to 0 ° C, and the reaction solution was added dropwise 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 ((4-cyclopropylphenoxy)(4-nitrophenoxy)phosphonyl)-L-alanine Propyl ester (1.48 g, 67%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.00-8.28 (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 ((4-cyclopropylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl) Preparation of -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl 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 (2.70 mL), and a solution of ^t BuMgCl (1M, 2.1 mL, 2.10 mmol) was added dropwise to the above solution at room temperature. Under stirring for 10 minutes, ((4-cyclopropylphenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester (938 mg, 2.09) was added dropwise to the reaction. A solution of mmol in 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 ((4-cyclopropyl Phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- 4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl ester (62 mg, 10%, epimer ratio: S _P /R _P =3.1:1) .

_{^{1 H NMR (400MHz, 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 =18.4Hz, 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),1.17-1.91(m , 1H), 1.15-1.39 (m, 12H), 0.88-0.98 (m, 2H), 0.55-0.65 (m, 2H); ³¹ P NMR (162MHz, CDCl ₃ ): δ 4.05, 3.58; MS m/z (ESI): 570.1 [M+H] ⁺ .

Embodiment 5

Preparation of the first step (4-bromo-2-methylphenoxy)(tri-butyl)dimethyl decane

4-bromo-2-methylphenol (2.1 g, 11 mmol) and imidazole (2.2 g, 33 mmol) dissolved in DMF (10 mL), cooled to 0 ° C, <RTI ID=0.0> ) 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 EtOAcqqqqHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH Phenoxy)(tri-butyl)dimethyl decane (2.4 g, 73%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 7.05 (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).

Preparation of the third step of the third-butyl (4-cyclopropyl-2-methylphenoxy) dimethyl decane

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)(tri-butyl)dimethyl decane 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 disappearance of the starting material was observed by LC-MS. The mixture was diluted with EtOAc (50 mL) and water (20 mL). The organic layer was washed with water (40 mL) and brine (40 mL) : PE) The title compound was obtained as the title compound: mp.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 6.65 (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 is the preparation of 4-cyclopropyl-2-methylphenol

To a flask of tert-butyl(4-cyclopropyl-2-methylphenoxy)dimethyl decane (1.1 g, 85% purity, 4 mmol) was added tetrabutylammonium fluoride (1M in THF, 12 mL, 12 mmol). 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 (HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH (0.64 g, crude yield 80%).

^{1 H NMR (400MHz, DMSO- d6} ): δ 8.98 (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).

Preparation of the fourth step ((4-cyclopropyl-2-methylphenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester

4-Nitrophenyl dichlorophosphate (900 mg, 3.6 mmol) was dissolved in CH ₂ Cl ₂ (7.5 mL), then cooled to -78 ° C, 4-cyclopropyl-2-methylphenol (600 mg, 4.0 mmol) and TEA (0.39g, 3.9mmol) in CH ₂ Cl ₂ (7.5mL) was added dropwise to the above solution in ten minutes, the reaction was stirred at to room temperature. The above solution was added dropwise to a solution of L-alanine isopropyl ester hydrochloride (600 mg, 3.6 mmol) in CH ₂ Cl ₂ (7.5 mL), then EtOAc (EtOAc) The reaction system was dropped into the minute. The reaction was stirred at 0<0>C for 1 h. EtOAc (EtOAc) (EtOAc)EtOAc. :3) The title compound ((4-cyclopropyl-2-methylphenoxy)(4-nitrophenoxy)phosphonyl)-L-alanine isopropyl ester (550 mg, 33%) .

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.22 (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).

The fifth step ((4-cyclopropyl-2-methylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1) Preparation of (2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) , 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. After stirring at room temperature for 10 minutes, ((4-cyclopropyl-2-methylbenzene) A solution of (4-nitrophenoxy)phosphonium)-L-alanine isopropyl 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 give the title compound ((4-cyclopropyl-2-methylphenoxy) (yield: CH ₂ Cl ₂ : MeOH = 50:1 to 10:1). ((2R,3R,4R,5R)-5-(2,4-Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran 2-yl)methoxy)phosphonium)-L-alanine isopropyl ester (29 mg, 9%, epimer ratio: S _P /R _P =9.3:1).

_{^{1 H NMR (400MHz, CD 3}} OD): δ 7.57 (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

Preparation of the first step 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) ⁱ PrOH (10 mL) was added dropwise EtOAc (EtOAc m. The reaction was carried out at 80 ° C for 30 minutes under microwave. The reaction was carried out in parallel. Three batches were prepared, and the reaction liquids of the three reactions were cooled, combined, diluted with EtOH, filtered through celite, concentrated, and chromatographed (eluent: PE: EA = 50:1) to give the title. Compound 3-(4-(Benzyloxy)phenyl)-propanyl (400 mg, 56%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 7.32 (m, 7H), 6.91 (d, J = 8.8Hz, 2H), 4.97 (m, 4H), 4.68 (t, J = 6.4Hz, 2H), 4.08 (m, 1H).

The second step is the preparation of 4-(oxabutyl-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 of a short hydrazine column and concentration of the filtrate gave the title compound 4-(m-butyl-butan-3-yl) phenol (250 mg, 100%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 7.29 (d, J = 8.4Hz, 2H), 6.83 (d, J = 8.4Hz, 2H), 5.05 (m, 2H), 4.74 (m, 2H), 4.19 (m, 1H).

Preparation of isopropyl ((4-nitrophenoxy)(4-(oxabutyl-3-yl)phenoxy)phosphonium)-L-alanine isopropyl ester

4-Nitrophenyl dichlorophosphate (427 mg, 1.67 mmol) was dissolved in CH ₂ Cl ₂ (6 mL), cooled to -78 ° C, 4-(ethylidene-3-yl)phenol (250 mg, 1.67 mmol) ) and TEA (233 μ L, 1.67mmol) in CH ₂ Cl ₂ (3mL) was added dropwise into the reaction at room temperature for 40 minutes. Then cooled to -78 ℃, was slowly added dropwise successively L- alanine isopropyl ester hydrochloride (280mg, 1.67mmol) in CH CH ₂ Cl ₂ (2.5mL) and a solution of TEA (466 μ L, 3.34mmol) of ₂ Cl ₂ (2 mL) solution, 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 ((4- nitrophenoxy) (4-(m- </RTI> -L-isopropyl propyl acrylate (540 mg, 70%). _{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.11 (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 (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-) Preparation of 4-methyltetrahydrofuran-2-yl)methoxy)(4-(oxabutyl-3-yl)phenoxy)phosphonium)-L-alanine isopropyl ester

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, ((4-nitrophenoxy)(4-(oxabutan-3-yl)phenoxy)phosphonium)-L-alanine was added dropwise to the reaction. A solution of the propyl ester (542 mg, 1.17 mmol) in THF (2 mL). Then cooled to room temperature, methanol (2mL) The reaction was quenched, concentrated to column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 20: 1) under reduced pressure to give the title compound ((((2R, 3R , 4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl) Methoxy)(4-(indolyl-3-yl)phenoxy)phosphonium)-L-alanine isopropyl ester (35 mg, 10%, a pair of epimers ratio S _P /R _P =2.5:1).

_{^{1 H NMR (400MHz, CD 3}} OD): δ 7.51 (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

Preparation of the first step 4-(cyclopropylmethyl)phenol

To a solution of 4-hydroxy-phenylcyclopropyl ketone (4.2 g, 26 mmol) in THF (15 mL), EtOAc (EtOAc,EtOAc. After an hour, boron trifluoride etherate (0.32 mL, 2.6 mmol) was then added and stirring was continued for 1 hour. After the TLC reaction was completed, the reaction mixture was poured into ice water, and the organic phase was separated, and the organic phase was dried over anhydrous magnesium sulfate, and concentrated, and the column chromatography (eluent: PE:EtOAc = 20:1) (Cyclopropylmethyl)phenol (3.7 g, 96%).

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.13 (d, J = 8.4 Hz, 2H), 6.80 (d, J = 8.4 Hz, 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).

Preparation of the second step ((4-(cyclopropylmethyl)phenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester

Dry ice-acetone (-78 ° C), to a solution of 4-nitrophenyl dichlorophosphate (1.50 g, 5.86 mmol) in CH ₂ Cl ₂ (30 mL) salt (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 EtOAcqqqqqHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH Propylmethyl)phenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester (180 mg, 6.7%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.17 (m, 2H), 7.32 (m, 2H), 7.l7 (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] ⁺ .

The third step ((4-(cyclopropylmethyl)phenoxy)(((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)phosphonium) isopropylamine isopropyl ester

1-((2R,3R,4R,5S)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)- The 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, (4-(cyclopropylmethyl)phenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester (180 mg, A solution of 0.39 mmol of 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. The residue was diluted with a mixture of CHCl ₃ and ⁱ PrOH (v: v = 3:1, 20 mL) washed with water 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 ((4- (cyclopropylmethyl Phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxyl 4-Methyltetrahydrofuran-2-yl)methoxy)phosphonium) isopropyl isopropyl ester (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

Preparation of the first step ((4-nitrophenoxy)(4-(phenylethynyl)phenoxy)phosphonium)-L-alanine isopropyl ester

To a solution of 4-nitrophenyl dichlorophosphate (924 mg, 3.61 mmol) in CH ₂ Cl ₂ (10 mL), dry-ice-acetone (-78 ° C) 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, slowly turn dropwise L- alanine isopropyl ester hydrochloride (605mg, 3.61mmol) in CH ₂ Cl ₂ (5mL) and a solution of TEA (1.67mL, 7.22mmol) in CH ₂ Cl ₂ (5mL) was added dropwise completed, Rin slowly to room temperature and stir overnight. The reaction mixture was concentrated. EtOAc was evaporated. EtOAcjjjjjjjjjjjj The title compound ((4-nitrophenoxy)(4-(phenylethynyl)phenoxy)phosphonium)-L-alanine isopropyl ester (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 (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- Preparation of 4-methyltetrahydrofuran-2-yl)methoxy)(4-(phenylethynyl)phenoxy)phosphonium)-L-alanine isopropyl ester

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 (4-nitrophenoxy)(4-(phenylethynyl)phenoxy)phosphonium)-L-alanine isopropyl to the reaction. A solution of the ester (102 mg, 0.200 mmol) in THF (1 mL). 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 ((((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) Phosphonic acid)-L-isopropyl isopropyl 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

Preparation of the first step 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 The title compound: 4-(cyclopropylethynyl)phenol (3.50 g, 97%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 7.28 (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).

Preparation of the second step ((4-(cyclopropylethynyl)phenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester

4-Nitrophenyl dichlorophosphate (2.50 g, 9.77 mmol) was dissolved in CH ₂ Cl ₂ (20 mL), cooled to -78 ° C, 4-(cyclopropylethynyl)phenol (1.55 g, 9.77 mmol) And 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 minutes. Then cooled to -78 ℃, was slowly added dropwise successively L- alanine isopropyl ester hydrochloride (1.64g, 9.77mmol) in CH ₂ Cl ₂ (5mL) and a solution of TEA (2.72mL, 19.5mmol) in CH ₂ cl ₂ (7mL) 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 ((4-(cyclopropylethynyl)phenoxy)(4-nitrophenoxy)phosphonyl)-L - Isopropyl propylamine (3.30 g, 72%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.24 (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] ⁺ .

The third step ((4-(cyclopropylethynyl)phenoxy)(((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)phosphonium)-L-alanine isopropyl ester

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 ((4-(cyclopropylethynyl)phenoxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropylate was added dropwise 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 to column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 30: 1) under reduced pressure to give the title compound ((4- (cyclopropylmethyl Ethyl ethyl)phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro- 3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl ester (45 mg, 38%, epimer ratio S _P /R _P = 3.4:1).

_{^{1 H NMR (400MHz, CD 3}} OD): δ 7.50 (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

Preparation of the first step 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. Days, LC-MS detected the disappearance of raw materials. The reaction mixture was poured into water (200 mL), EtOAc (EtOAc (EtOAc) The title compound 4-(propylthio)phenol (0.9 g, 27%) was obtained.

^{1 H NMR (400MHz, DMSO- d} 6): δ 9.52 (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 (4-nitrophenoxy)(4-(propylthio)phenoxy)phosphonium)-L-alanine isopropyl ester

4-Nitrophenyl dichlorophosphate (1.3 g, 5 mmol) was dissolved in CH ₂ Cl ₂ (10 mL) in EtOAc. 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 Drop into the reaction system. The reaction was stirred at 0&lt;0&gt;C for 1 h. EtOAc (EtOAc) (EtOAc)EtOAc. The title compound ((4-nitrophenoxy)(4-(propylthio)phenoxy)phosphonyl)-L-alanine isopropyl ester (1.5 g, 60%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.16 (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 (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- Preparation of 4-methyltetrahydrofuran-2-yl)methoxy)(4-(propylthio)phenoxy)phosphonium)-L-alanine isopropyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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, ((4-nitrophenoxy) (4- A solution of (propylthio)phenoxy)phosphonium)-L-alanine isopropyl ester (484 mg, 1 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction was concentrated to half take column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound ((((2R, 3R, 4R, 5R) -5- (2 ,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(propyl) Isopropyl thio)phenoxy)phosphonium)-L-alanine (66 mg, 22%, epimer ratio: S _P /R _P = 3.5:1).

_{^{1 H NMR (400MHz, CD 3}} OD): δ 7.63 (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

Preparation of the first step 4-(tert-butylthio)phenol

To the third 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-(t-butylthio)phenol (3.0 g, 65%) was obtained.

^{1 H NMR (400MHz, DMSO- d} 6): δ 9.76 (s, 1H), 7.28 (d, J = 8.8Hz, 2H), 6.77 (d, J = 8.8Hz, 2H), 1.19 (s, 9H) .

Second step (Preparation of (4-nitrophenoxy)(4-(t-butylthio)phenoxy)phosphonium)-L-alanine isopropyl ester

4-Nitrophenyl dichlorophosphate (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 above solution was added dropwise to a solution of L-alanine isopropyl ester hydrochloride (2.5 g, 15 mmol) in CH ₂ Cl ₂ (30 mL), then EtOAc (EtOAc) Drop into the reaction system. The reaction was stirred at 0&lt;0&gt;C for 1 h. EtOAc (EtOAc) (EtOAc)EtOAc. :4) The title compound ((4-nitrophenoxy)(4-(t-butylthio)phenoxy)phosphonium)-L-alanine isopropyl ester (3.0 g, 40%) ).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.23 (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 (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- Preparation of 4-methyltetrahydrofuran-2-yl)methoxy)(4-(t-butylthio)phenoxy)phosphonium)-L-alanine isopropyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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, ((4-nitrophenoxy) (4- A solution of (t-butylthio)phenoxy)phosphonium)-L-alanine isopropyl ester (500 mg, 1 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated after column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound ((((2R, 3R, 4R, 5R) -5- (2,4 -dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(t-butyl) Isopropyl thio)phenoxy)phosphonium)-L-alanine (126 mg, 41%, epimer ratio: S _P /R _P = 2.2:1).

_{^{1 H NMR (400MHz, CD 3}} OD): δ 7.66 (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 - 1.17 (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

Preparation of ethyl (4-methoxybenzyl) sulfane in the first step

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}} ): δ 7.26 (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).

The second step is the 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 = 9:1~6:4) The title compound ethyl (4-methoxybenzyl)sulfane (2.5 g, 67%) was obtained.

^{1 H NMR (400MHz, DMSO- d} 6): δ 9.31 (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).

Step 3: Preparation of (4-nitrophenoxy)(4-((ethylthio)methyl)phenoxy)phosphonium)-L-alanine isopropyl ester

4-Nitrophenyl dichlorophosphate (3.5 g, 13.5 mmol) was dissolved in CH ₂ Cl ₂ (27 mL) in EtOAc. EtOAc (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 solution was added dropwise L- alanine isopropyl ester hydrochloride was cooled at 0 ℃ (2.3g, 13.5mmol) in CH ₂ Cl ₂ (27mL) solution, and then TEA (3.0g, 30mmol) in 5 minutes The reaction system was dropwise added thereto, and the reaction was slowly warmed to room temperature and stirred overnight. After concentrating the solution, EtOAc (EtOAc (EtOAc)EtOAc. -Nitrophenoxy)(4-((ethylthio)methyl)phenoxy)phosphonium)-L-alanine isopropyl ester (1.7 g, 25%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.23 (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 (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-) Preparation of 4-methyltetrahydrofuran-2-yl)methoxy)(4-((ethylthio)methyl)phenoxy)phosphonium)-L-alanine isopropyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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. After stirring at room temperature for 10 minutes, ((4-nitrophenoxy) (4- A solution of ((ethylthio)methyl)phenoxy)phosphonium)-L-alanine isopropyl ester (480 mg, 1 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated after column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound ((((2R, 3R, 4R, 5R) -5- (2,4 -dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-((ethylthio) Methyl)phenoxy)phosphonium)-L-alanine isopropyl ester (150 mg, 50%, epimer ratio: S _P /R _P = 2.2:1).

_{^{1 H NMR (400MHz, CD 3}} OD): δ 7.63 (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

Preparation of the first step ((4-nitrophenoxy)(4-(trimethylsulfonyl)phenoxy)phosphonium)-L-alanine isopropyl ester

4-Nitrophenyl dichlorophosphate (1.02 g, 4 mmol) was dissolved in CH ₂ Cl ₂ (8 mL) in room temperature, then cooled to -78 ° C, 4-(trimethylmethyl) phenol (750 mg) A solution of 4.5 mmol of 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 under stirring. The above solution was added dropwise to a solution of L-alanine isopropyl ester hydrochloride (690 mg, 4 mmol) in CH ₂ Cl ₂ (8 mL), then EtOAc (EtOAc) reaction system. The reaction was stirred at 0&lt;0&gt;C for 1 h, EtOAc (EtOAc) (EtOAc)EtOAc. The title compound ((4-nitrophenoxy)(4-(trimethylcarbinyl)phenoxy)phosphonium)-L-alanine isopropyl ester (1.2 g, 60%) was obtained.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.13 (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 (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- Preparation of 4-methyltetrahydrofuran-2-yl)methoxy)(4-(trimethylsulfonyl)phenoxy)phosphonium)-L-alanine isopropyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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, ((4-nitrophenoxy) (4- A solution of (trimethylindenyl)phenoxy)phosphonium)-L-alanine isopropyl ester (480 mg, 1.0 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction was concentrated to half take column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound ((((2R, 3R, 4R, 5R) -5- (2 ,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(trimethyl) Phenyloxy)phenoxy)phosphonium)-L-alanine isopropyl ester (38 mg, 13%, epimer ratio: S _P /R _P >10:1).

_{^{1 H NMR (400MHz, CD 3}} OD): δ 7.49 (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), 1.10-1.14 (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

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

4-Nitrophenyl dichlorophosphate (2.0 g, 8.0 mmol) dissolved in CH ₂ Cl ₂ (16 mL), cooled to -78 ° C, 2,3-dihydro-1H-indole-5-ol (1.2 g A solution of 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 the mixture was allowed to warm to room temperature under stirring. The above solution was added dropwise to a solution of L-alanine isopropyl ester hydrochloride (1.35 g, 8 mmol) in CH ₂ Cl ₂ (16 mL), EtOAc (EtOAc) Drop into the reaction system. The reaction was stirred at 0&lt;0&gt;C for 1 h, EtOAc (EtOAc) (EtOAc)EtOAc. The title compound (((2,3-dihydro-1H-indol-5-yl)oxy)(4-nitrophenoxy)phosphonyl)-L-alanine isopropyl ester (1.9 g) , 50%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.15 (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 (((2,3-dihydro-1H-indol-5-yl)oxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4- Preparation of dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) , 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 (((2,3-dihydro-1H-indole). A solution of -5-yl)oxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester (900 mg, 2 mmol) in anhydrous THF (6 mL) was added dropwise over five minutes. The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction was concentrated to half take column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound (((2,3-dihydro-inden-5-yl -1H- )oxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- 4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl ester (40 mg, 7%, epimer ratio: S _P /R _P =3.3:1) .

_{^{1 H NMR (400MHz, CD 3}} OD): δ 7.61 (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 (162MHz, CD 3 OD): δ 3.96,3.90; MS m / z (ESI): 570.2 [m + H] +.

Example fifteen

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

4-Nitrophenyl dichlorophosphate (600 mg, 2.4 mmol) was dissolved in CH ₂ Cl ₂ (5 mL) in room temperature, then cooled to -78 ° C, 5,6,7,8-tetrahydronaphthalene - 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 with stirring. The above solution was added dropwise to a solution of L-alanine isopropyl ester hydrochloride (400 mg, 2.4 mmol) in CH ₂ Cl ₂ (5 mL), EtOAc (EtOAc) Into the reaction system. The reaction was stirred at 0&lt;0&gt;C for 1 h. EtOAc (EtOAc) (EtOAc)EtOAc. The title compound ((4-nitrophenoxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphonium)-L-alanine isopropyl ester (480 mg) , 44%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.16 (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 (((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- Preparation of 4-methyltetrahydrofuran-2-yl)methoxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphonium)-L-alanine isopropyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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 ((4-nitrophenoxy) (5 A solution of 6,6,8-tetrahydronaphthalen-2-yl)oxy)phosphonium)-L-alanine isopropyl ester (480 mg, 1 mmol) in dry THF (3 mL) The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction was concentrated to half take column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound (((2,3-dihydro-inden-5-yl -1H- )oxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- 4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl ester (25 mg, 9%, epimer ratio: S _P /R _P =4.0:1) .

_{^{1 H NMR (400MHz, CD 3}} OD): δ 7.59 (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 (((2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxy)(4-nitrophenoxy)phosphonium)-L-propylamine Preparation of isopropyl acid ester

4-Nitrophenyl dichlorophosphate (600 mg, 2.4 mmol) was dissolved in CH ₂ Cl ₂ (5 mL) in room temperature, then cooled to -78 ° C, 2,3-dihydrobenzo[b][ 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 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) cooled at 0 ° C, then TEA (500 mg, 5 mmol) Into the reaction system. The reaction was stirred at 0&lt;0&gt;C for 1 h. EtOAc (EtOAc) (EtOAc)EtOAc. The title compound (((2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxy)(4-nitrophenoxy)phosphonium)-L- Isopropyl propylamine (580 mg, 53%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.25 (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 (((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)phosphonium)-L- Preparation of isopropyl propylamine

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) , 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 (((2,3-dihydrobenzo[b] [1,4] Dioxin-6-yl)oxy)(4-nitrophenoxy)phosphonium)-L-alanine isopropyl ester (580 mg, 1.2 mmol) in THF (3 mL) The solution was instilled within five minutes. The reaction was stirred at 55 &lt;0&gt;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) and purification afforded the title compound (((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 Isopropylphosphonium)-L-alanine isopropyl ester (20 mg, 6%, epimer ratio: S _P /R _P = 1.9:1).

_{^{1 H NMR (400MHz, 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

Preparation of the first step (2S)-2-((4-nitrophenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester

Phenyl dichlorophosphate (1.0 g, 4.7 mmol) was dissolved in CH ₂ Cl ₂ (10 mL) in EtOAc. EtOAc EtOAc (EtOAc) A solution of 5.4 mmol) in CH ₂ Cl ₂ (10 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)-2-aminopropylthiocarboxylic acid S-isopropyl ester hydrochloride (1.2 g, 4.7 mmol) in CH ₂ Cl ₂ (10 mL). Then, TEA (1.0 g, 10 mmol) was dropped into the reaction system over 5 minutes. The reaction was stirred at 0&lt;0&gt;C for 1 h. EtOAc (EtOAc) (EtOAc)EtOAc. :1) The title compound 2S)-2-((4-nitrophenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl (ester (1.03 g) , 50%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.16 (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 (2S)-2-((((((((((((((((((((((((((())))))) Of -Fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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, (2S)-2-(((4-nitro) A solution of phenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (424 mg, 1.0 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction was concentrated to half take column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound (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)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (66 mg, 24%, epimer ratio: S _P /R _P = 2.2:1).

_{^{1 H NMR (400MHz, 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 = 18.8Hz, 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)-2-((Tertiary-butoxycarbonyl)amino)propylthiocarboxylic acid S-methyl ester

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 ( 4.0 g, 30 mmol) was injected into the above solution. 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 (jjjjjjjjjjjjjj Acid S-methyl ester (3.4 g, 67%).

^{1 H NMR (400MHz, DMSO- d} 6): δ 4.97 (s, 1H), 4.40 (q, J = 7.2Hz, 1H), 2.29 (s, 3H), 1.46 (s, 9H), 1.37 (d, 3H).

Preparation of (S)-2-aminopropylthiocarboxylic acid S-methyl ester hydrochloride

Add HCl (4N in dioxane, 20 mL) to a flask containing (S)-2-((t-butoxycarbonyl)amino)propylthiocarboxylic acid S-methyl ester (3.4 g, 16 mmol). The solution was stirred at room temperature for three hours, and the disappearance of the starting material was confirmed by LC-MS. The solvent was concentrated to give the title compound (S)-2-aminopropyl thio carboxylic acid S-methyl ester hydrochloride (2.5 g, crude yield 100%).

^{1 H NMR (400MHz, DMSO- d} 6): δ 8.59 (s, 3H), 4.29 (q, J = 7.2Hz, 1H), 2.37 (s, 3H), 1.45 (d, J = 7.2Hz, 3H) .

Preparation of (2S)-2-((4-nitrophenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-methyl ester

Phenyl dichlorophosphate (3.15 g, 15 mmol) was dissolved in CH ₂ Cl ₂ (30 mL) in EtOAc. EtOAc (EtOAc) A solution of 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)-2-aminopropylthiocarboxylic acid S-methyl ester hydrochloride (2.3 g, 15 mmol) in CH ₂ Cl ₂ (30 mL). Then, TEA (3.3 g, 33 mmol) was dropped into the reaction system over 5 minutes. The reaction was stirred at 0&lt;0&gt;C for 1 h. EtOAc (EtOAc) (EtOAc)EtOAc. :1) The title compound (2S)-2-((4-nitrophenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-methyl ester (2.7 g, 45%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.23 (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 fourth step (2S)-2-((((((((((((((((((((((())))))) Of -fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-methyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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, (2S)-2-(((4-nitro) A solution of the phenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-methyl ester (400 mg, 1 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (3 mL) The reaction mixture was concentrated after column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound (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)phosphonium)amino)propylthiocarboxylic acid S-methyl 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 (162MHz, CD 3 OD): δ 3.83,3.54; MS m / z (ESI): 518.0 [m + H] +.

Example 19

Preparation of S-(Tertiary-Butyl) Ester of First Step (S)-2-((Tertiary-Butoxycarbonyl)Amino)propylthiocarboxylic Acid

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. Thiol (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). The title compound (S)-2-((tris-butoxycarbonyl)amino)propylsulfide was obtained by column chromatography (eluent: PE:EtOAc: 20:1 to 6:1). S-(T-butyl) carboxylic acid (2.2 g, 17%).

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

Preparation of (S)-2-aminopropylthiocarboxylic acid S-tert-butyl ester hydrochloride

(S)-2-((Terti-butoxycarbonyl)amino)propylthiocarboxylic acid S-(T-butyl) ester (2.2 g, 8.5 mmol) dissolved in 4N hydrochloric acid in dioxane In (20 mL), the reaction was stirred at 30 ° C for two hours. The disappearance of the starting material was observed by LC-MS, and the title compound (S)-2-aminopropylthio carboxylic acid S-t-butyl ester hydrochloride (1.7 g, crude yield 100%).

^{1 H NMR (400MHz, DMSO- d} 6): δ 8.59 (s, 3H), 4.14 (m, 1H), 1.48 (s, 9H), 1.43 (d, J = 7.2Hz, 3H).

Preparation of (2S)-2-((4-nitrophenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-t-butyl ester

Phenyl dichlorophosphate (1.7 g, 8.0 mmol) was dissolved in CH ₂ Cl ₂ (15 mL) in EtOAc. EtOAc (EtOAc) A solution of g, 9.0 mmol) in 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)-2-aminopropylthiocarboxylic acid S-tert-butyl ester hydrochloride (1.7 g, 8.5 mmol) in CH ₂ Cl ₂ (15 mL) at 0 ° C. Then, TEA (1.8 g, 18 mmol) was added dropwise to the reaction system over 5 minutes. The reaction was stirred at 0&lt;0&gt;C for 1 h. EtOAc (EtOAc) (EtOAc)EtOAc. : 4) The title compound (2S)-2-((4-nitrophenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-t-butyl ester (1.9) was obtained. g, 53%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.22 (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 (2S)-2-((((((((((((((((((((((())))))) Of -Fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-tert-butyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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, (2S)-2-(((4-nitro) A solution of phenoxy)(phenoxy)phosphonyl)amino)propylthiocarboxylic acid S-tert-butyl ester (440 mg, 1.0 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated after column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound (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)phosphonium)amino)propylthiocarboxylic acid S-tert-butyl ester (134 mg, 49%, epimer ratio: S _P /R _P = 2.2:1).

_{^{1 H NMR (400MHz, CD 3}} OD): δ 7.64-7.66 (d, J = 8.4Hz, 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} (162MHz, CD 3 OD): δ 3.49; MS m / z (ESI): 560.0 [m + H] +.

Example twenty

Preparation of the first step (2S)-2-((4-nitrophenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-cyclohexyl ester

Phenyl dichlorophosphate (1.7 g, 8.0 mmol) was dissolved in CH ₂ Cl ₂ (15 mL) in EtOAc. EtOAc (EtOAc) A solution of g, 9.0 mmol) in 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)-2-aminopropylthiocarboxylic acid S-cyclohexyl ester hydrochloride (1.8 g, 8.0 mmol) in CH ₂ Cl ₂ (15 mL). TEA (1.8 g, 18 mmol) was then added dropwise to the reaction system over 5 minutes. The reaction was stirred at 0&lt;0&gt;C for 1 h. EtOAc (EtOAc) (EtOAc)EtOAc. : 4) The title compound (2S)-2-((4-nitrophenoxy)(phenoxy)phosphonium)amino)propyl thiocarboxylate S-cyclohexyl ester (2.5 g, 70%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.22 (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 (2S)-2-((((((((((((((((((((((((((())))))) Of -fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-cyclohexyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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, (2S)-2-(((4-nitro) A solution of phenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-cyclohexyl ester (424 mg, 1.0 mmol) in THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated after column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound (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)phosphonium)amino)propylthiocarboxylic acid S-cyclohexyl ester (122 mg, 42%, epimer ratio: S _P /R _P = 2.2:1).

_{^{1 H NMR (400MHz, 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 = 18.8Hz, 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 (2S)-2-((4-nitrophenoxy)(4-cyclopropylphenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester preparation

Dichloro-4-nitrophenyl phosphate (1.75g, 6.75mmol) was dissolved in CH ₂ Cl ₂ (15mL), cooled to -78 deg.] C, 4- cyclopropyl-phenol (1.0g, 7.5mmol) and TEA ( A solution of 0.75 g, 7.5 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 added dropwise to 0 ° C to cool (S). -2-amino-propylthio acid S- isopropyl ester hydrochloride (1.3g, 6.75mmol) in (15mL) solution of CH ₂ Cl _2, followed by TEA (1.5g, 15mmol) was added dropwise to the reaction system, The mixture was stirred at 0&lt;0&gt;C for 1 h. EtOAc was evaporated. Column chromatography (eluent: PE: EtOAc = 9:1 to 1.5:1) afforded the title compound (2S)-2-(((4-nitrophenoxy) (4-cyclopropylphenoxy) Phosphonic acid)amino)propylthiocarboxylic acid S-isopropyl ester (2.1 g, 70%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.20-8.23 (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 (2S)-2-((((((((((((((((((((((((((())))))) -fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-cyclopropylphenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester Preparation

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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, (2S)-2-(((4-nitrobenzene) A solution of (4-cyclopropylphenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (464 mg, 1.0 mmol) in THF (3 mL) was added dropwise over five minutes . The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated after column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound (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) 4-cyclopropylphenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (60 mg, 20%, epimer ratio S _P /R _P =4.7:1 ).

_{^{1 H NMR (400MHz, 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 = 19.2Hz, 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 (2S)-2-(((4-nitrophenoxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphonium)amino)propyl) Preparation of S-isopropyl thiocarboxylate

4-Nitrophenyl dichlorophosphate (1.75 g, 7.0 mmol) was dissolved in CH ₂ Cl ₂ (15 mL) and cooled to -78 ° C, 5,6,7,8-tetrahydro-2-naphthol ( A solution of 1.4 g, 8.0 mmol) and 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 added dropwise. cooled at 0 ℃ to (S) -2- aminopropyl thiocarboxylic acid S- isopropyl ester hydrochloride (1.4g, 8.0mmol) in CH ₂ Cl ₂ (15mL) solution, followed by TEA (1.5g The reaction mixture was added dropwise, and the mixture was stirred at EtOAc EtOAc. Column chromatography (eluent: PE: EtOAc = 9:1 to 1.5:1) afforded the title compound (2S)-2-(((4-nitrophenoxy) ((5,6,7,8,8) -tetrahydronaphthalen-2-yl)oxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (2.2 g, 65%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.20-8.24 (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 (2S)-2-((((((((((((((((((((((((((())))))) -fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphonium)amino)propyl Preparation of S-Isopropyl Carboxylic Acid

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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, (2S)-2-(((4-nitrobenzene) Oxy)((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (480 mg, 1.0 mmol) A solution of THF (3 mL) was added dropwise over five minutes. The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated after column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound (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) (5,6,7,8-tetrahydronaphthalen-2-yl)oxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (148 mg, 50%, epimer The ratio is S _P /R _P =2.7:1).

_{^{1 H NMR (400MHz, 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 = 20.4Hz, 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 (2S)-2-(((4-nitrophenoxy)(4-(propylthio)phenoxy)phosphonium)amino)propyl thiocarboxylic acid S-isopropyl Preparation of base ester

4-Nitrophenyl dichlorophosphate (1.75 g, 7.0 mmol) was dissolved in CH ₂ Cl ₂ (15 mL), cooled to -78 ° C, 4-(propylthio) phenol (1.4 g, 8.0 mmol) 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 added dropwise to 0 ° C to cool. (S) -2- aminopropyl thiocarboxylic acid S- isopropyl ester hydrochloride (1.4g, 8.0mmol) in CH ₂ Cl ₂ (15mL) solution, followed by TEA (1.5g, 15mmol) by The reaction mixture was added dropwise, and the mixture was stirred for 1 hr. Column chromatography (eluent: PE: EtOAc = 9:1 to 1.5:1) gave the title compound (2S)-2-(((4-nitrophenoxy) (4-(propylthio)) Phenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (2.5 g, 70%).

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 8.21-8.24 (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 (2S)-2-((((((((((((((((((((((((((())))))) -fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(4-(propylthio)phenoxy)phosphonium)amino)propylthiocarboxylic acid S-iso Preparation of propyl ester

1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4 (1H) 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, (2S)-2-(((4-nitrobenzene) a solution of (4-(propylthio)phenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (500 mg, 1.0 mmol) in THF (3 mL) Drip inside. The reaction was stirred at 55 &lt;0&gt;C overnight, cooled to rt and EtOAc (EtOAc) The reaction mixture was concentrated after column chromatography _{(eluent: CH 2 Cl 2: MeOH =} 50: 1 ~ 10: 1) to give the title compound (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) (4-(propylthio)phenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (141 mg, 46%, epimer ratio S _P /R _P =1.5:1).

_{^{1 H NMR (400MHz, 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 = 17.2Hz, 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

Preparation of the first step of 4-cyclopropyl phenyl dichlorophosphate

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-dichloro-phosphoric acid phenyl ester (4.4g, 91%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.18-7.14 (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.

Preparation of the second step ((4-cyclopropylphenoxy)(perfluorophenoxy)phosphonium)-L-alanine isopropyl 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 mmol) was slowly added dropwise under N ₂ protection. After stirring for 15 minutes, a solution of 4-cyclopropylphenyl dichlorophosphate (3.7 g, 14.82 mmol) dissolved in CH ₂ Cl ₂ (5 mL) was slowly added dropwise, stirred for 30 minutes and then slowly warmed to 0 ° C. Stirring was continued for 1 hour at this temperature.

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, mjjjjjjjjjjjjjjjjjj Indyl)-L-isopropyl isopropyl ester (6.03 g, 82%, epimer ratio: S _P /R _P = 1:1).

^{1 H NMR (400MHz, DMSO- d} 6) δ 7.16-7.08 (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 (S)-(4-cyclopropylphenoxy)(perfluorophenoxy)phosphonium)-L-alanine isopropyl ester

The (2-cyclopropylphenoxy)(perfluorophenoxy)phosphonium)-L-alanine isopropyl 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 (EtOAc) Phenoxy)phosphonium)-L-alanine isopropyl ester (2.99 g, 40%, HPLC purity: 97%).

^{1 H NMR (400MHz, DMSO- d} 6) δ 7.09 (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 = 6.8Hz, 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 (S)-(4-cyclopropylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidine-1 (2H) Preparation of isopropyl 4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine

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 ((S)-(4-cyclopropylphenoxy)(perfluorophenoxy)phosphonium)-L- dissolved in THF (3.6 mL) was slowly added dropwise. A solution of isopropyl propylamine (1.0 g, 2.03 mmol) was maintained at this temperature and stirred 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 obtained solid was recrystallized from CH ₂ Cl ₂ to give the title compound (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)phosphonium)-L-alanine isopropyl Base ester (680 mg, 70%, HPLC purity: 98%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 9.25 (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 (162MHz, CDCl 3}} ) δ 3.65; 19 F NMR (376MHz, CDCl 3) δ -161.79; MS m / z (ESI): 570.1 [m + H] +.

((R)-(4-cyclopropylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)- Of 4-(4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl 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 ((4-cyclopropylphenoxy)(perfluorophenoxy)phosphonium)-L-alanine isopropyl 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). The resulting crude product was column chromatographed _{^{(CH 2 Cl 2: i PrOH}} = 20: 1) to give the title compound ((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)phosphonium Base)-L-isopropyl isopropyl ester (118 mg, 24%, HPLC purity: 98%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 8.95 (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 (162MHz, CDCl ₃ ) δ 4.31; ¹⁹ F NMR (376MHz, CDCl ₃ ) δ -162.04; MS m/z (ESI): 570.1 [M +H] ⁺ .

Example twenty five

((S)-((2R,3R,4R,5R)-3-Ethyloxy-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)- 4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)(4-cyclopropylphenoxy)phosphonium)-L-alanine isopropyl ester

(S)-(4-cyclopropylphenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-) 4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)phosphonium)-L-alanine isopropyl ester (57 mg, 0.10 mmol) in THF (7 mL) To the solution were added ethyl hydrazine chloride (0.035 mL, 0.50 mmol), DMAP (61 mg, 0.50 mmol), and 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 ((S) -(((2R,3R,4R,5R)-3-ethoxycarbonyl-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro- 4-methyltetrahydrofuran-2-yl)methoxy)(4-cyclopropylphenoxy)phosphonium)-L-alanine isopropyl ester (45 mg, 74%, HPLC purity: 98%) ¹ H NMR (400 MHz, MeOD) δ 7.55 (d, J = 8.0 Hz, 1H), 7.06 (m, 4H), 6.03 (d, J = 18.4 Hz, 1H), 5.55 (d, J = 8.0 Hz, 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

Preparation of the first step ((S)-((2,3-dihydro-1H-indol-5-yl)oxy)(perfluorophenoxy)phosphonium)-L-alanine isopropyl ester

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) dichlorophosphate.

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. 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 salt had been added, and the solution was added dropwise 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) (EtOAc) (EtOAcjjjjjjj Phenoxy)phosphonium)-L-alanine isopropyl ester (14.5 g, 20%).

^{1 H NMR (400MHz, MeOD)} δ 7.20 (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 (S)-((2,3-dihydro-1H-indol-5-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)phosphonium)-L-alanine isopropyl ester Preparation

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 ((S)-((2,3-dihydro-1H-indol-5-yl)oxy) (perfluorophenoxy) dissolved in THF (50 mL) was slowly added dropwise. A solution of hydrazino)-L-alanine isopropyl ester (5.6 g, 11.36 mmol) was maintained at this temperature and stirred 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. sodium hydrogen sulfate (100 mL) and brine (100 mL). Column chromatography (eluent: CH ₂ Cl ₂ : ⁱ PrOH = 60:1 to 12:1) gave the title compound (S)-((2,3-dihydro-1H-indole-5-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)phosphonium)-L-alanine isopropyl ester (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

((S)-((2R,3R,4R,5R)-3-Ethyloxy-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)- 4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)(4-cyclopropylphenoxy)phosphonium)-L-alanine isopropyl ester

Take (S)-((2,3-dihydro-1H-indol-5-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)phosphonium)-L-alanine isopropyl ester (57mg Ethyl acetate (0.035 mL, 0.50 mmol), DMAP (61 mg, 0.50 mmol), and then stirred at room temperature for 3 hr. 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 ((S) -(((2R,3R,4R,5R)-3-ethoxycarbonyl-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro- 4-Methyltetrahydrofuran-2-yl)methoxy)(4-cyclopropylphenoxy)phosphonium)-L-alanine isopropyl ester (40 mg, 65%).

^{1 H NMR (400MHz, CDCl3)} δ 9.37 (s, 1H), 7.48 (d, J = 8.0Hz, 1H), 7.06 (d, J = 8.0Hz, 1H), 7.01 (s, 1H), 6.88 (d , J = 8.0Hz, 1H), 6.13 (d, J = 18.4Hz, 1H), 5,46 (d, J = 8.0Hz, 1H), 5.12 (dd, J = 20.4,8.0Hz, 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 ((S)-((5,6,7,8-tetrahydronaphthalen-2-yl)oxy)(perfluorophenoxy)phosphonium)-L-alanine isopropyl ester preparation

5,6,7,8-tetrahydronaphthalen-2-ol (23.0 g, 150 mmol) was weighed and dissolved in Et ₂ O (150 mL), and 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) dichlorophosphate.

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. The solution of dichlorophosphoric acid (5,6,7,8-tetrahydronaphthalen-2-yl) ester cooled at -78 ° C was slowly added dropwise thereto for about 2 hours. The solution slowly rose to zero degrees.

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 salt had been added, and the solution was added dropwise for half an hour, and the solution was stirred at room temperature overnight.

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 stirred in EtOAc (3 mL) EtOAc (EtOAc) (EtOAc) (Perfluorophenoxy)phosphonium)-L-alanine isopropyl ester (22.0 g, 28%).

^{1 H NMR (400MHz, MeOD)} δ 7.06 (d, J = 9.2Hz, 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 (S)-((5,6,7,8-tetrahydronaphthalen-2-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)phosphonium)-L-alanine isopropyl 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 ((S)-((5,6,7,8-tetrahydronaphthalen-2-yl)oxy) (perfluorobenzene) dissolved in THF (50 mL) was slowly added dropwise. A solution of oxy)phosphonium)-L-alanine isopropyl ester (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 (S)-((5,6,7,8-tetrahydronaphthalen-2-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)phosphonium)-L - Isopropyl propylamine (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 (162MHz, MeOD) δ 3.87; ¹⁹ F NMR (376 MHz, MeOD) δ -162.11; MS m/z (ES1): 584.0 [M+H] ⁺ .

Example twenty nine

((S)-((2R,3R,4R,5R)-3-Ethyloxy-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)phosphonium)-L-alanine isopropyl Base ester

Take (S)-((5,6,7,8-tetrahydronaphthalen-2-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)phosphonium)-L-alanine isopropyl ester ( 60 mg, 0.10 mmol) was dissolved in THF (7 mL). EtOAc (EtOAc (EtOAc,EtOAc) hour. Diluted with EtOAc, the organic phase washed successively with water, saturated brine, dried over anhydrous sodium sulfate, concentrated, column chromatography _{^{(eluent: CH 2 Cl 2: i PrOH}} = 60: 1 ~ 12: 1) to give the title Compound ((S)-(((2R,3R,4R,5R)-3-ethoxycarbonyl-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)phosphonium)-L-alanine Propyl ester (29 mg, 40%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 9.14 (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

Preparation of the first step ((S)-(4-(propylthio)phenoxy)(perfluorophenoxy)phosphonium)-L-alanine isopropyl 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 mixture was slowly warmed to room temperature and stirred overnight, and LCMS showed to give (4-(propylthio)phenoxy)dichlorodithioate.

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. The solution of (4-(propylthio)phenoxy)dichlorophosphoric acid dichloride 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.

Weigh pentafluorophenol (17.7g, 96mmol) was dissolved in CH ₂ Cl ₂ (120mL), was placed in an ice bath, was slowly added dropwise with stirring DIPEA (12.4g, 96mmol). The above solution was slowly dropped into a solution at 0 ° C to which L-alanine isopropyl ester salt 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) Propyl ester (10.9 g, 27%).

¹ H NMR (400 MHz, MeOD) δ 7.39 (m, 2H), 7.22 (m, 2H), 4.97 (m, 1H), 4.03 (m, 1H), 2.92 (t, J = 7.2 Hz, 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 (S)-(4-(propylthio)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)phosphonium)-L-alanine isopropyl ester

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)-(4-(propylthio))phenoxy)(perfluorophenoxy)phosphonium) dissolved in THF (25 mL) was slowly added dropwise. A solution of L-alanine isopropyl ester (3.32 g, 6.3 mmol) was maintained at this temperature and stirred 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 (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)phosphonium)-L-alanine isopropyl ester ( 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, MeOH) δ 3.89; ¹⁹ F NMR (376 MHz,MeOD) δ -161.87; MS m/z (ESI): 604.0 [M+H] ⁺ .

Embodiment 31

((S)-((2R,3R,4R,5R)-3-Ethyloxy-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)- 4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)(4-(propylthio)phenoxy)phosphonium)-L-alanine isopropyl ester

(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)phosphonium)-L-alanine isopropyl ester (60 mg, 0.10 mmol) dissolved in THF (7 mL), to the solution were added ethyl chlorobenzene (0.035 mL, 0.50 mmol), DMAP (61 mg, 0.50 mmol), and 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 ((S) -(((2R,3R,4R,5R)-3-ethoxycarbonyl-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro- 4-Methyltetrahydrofuran-2-yl)methoxy)(4-(propylthio)phenoxy)phosphonium)-L-alanine isopropyl ester (50 mg, 77%).

^{1 H NMR (400MHz, MeOD)} δ 7.53 (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 = 7.2Hz, 3H) ; 31 P NMR (162MHz, MeOD) δ 3.85; 19 F NMR (376MHz, MeOD) δ -157.38; MS m / z (ESI): 646.2 [m + H] +.

Example thirty-two

First step (S)-2-(((S)-(perfluorophenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester

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 dichlorophosphate.

(S)-2-Aminopropylthiocarboxylic acid S-isopropyl ester hydrochloride (16.5 g, 90 mmol) was dissolved in CH ₂ Cl ₂ (200 mL), and DIPEA (23.2 g, 180 mmol) was added. Stir well. The solution was slowly dropped into a cooled phenyl dichlorophosphate solution at -78 ° C, and the addition time was 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)-2-aminopropylthiocarboxylic acid S-isopropyl ester hydrochloride was added, and the mixture was dropwise added 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)-2-(((S)-(perfluorophenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (7.1 g) , 15%).

^{1 H NMR (400MHz, MeOD)} δ 7.42 (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 (376MHz, MeOD) δ -154.86 (d, J = 20.0 Hz, 2F), -162.92 (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)-2-(((S)-((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-) 4-fluoro-3-hydroxy-4-methylhydrofuran-2-yl)methoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester Preparation

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)-2-(((S)-(perfluorophenoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S was slowly added dropwise. A solution of isopropyl ester (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, crystals crystals eluted from ethyl ether (ethyl ether) 2,4-Dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methylhydrofuran-2-yl)methoxy)(phenoxy) Phosphonic acid) amino) S-isopropyl propyl sulfate (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, MeOH) δ 3.53; ¹⁹ F NMR (376 MHz, MeOD) δ -161.97; MS m/z (ESI): 546.0 [M +H] ⁺ .

Example thirty three

Acetic acid (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)phosphonyl)oxy)methyl)-4-methyltetrahydrofuran- 3-based ester

(S)-2-(((()))) 4-fluoro-3-hydroxy-4-methylhydrofuran-2-yl)methoxy)(phenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester (54 mg Ethyl acetate (0.035 mL, 0.50 mmol), DMAP (61 mg, 0.50 mmol), and then stirred at room temperature for 3 hr. Diluted with EtOAc, the organic phase washed successively with water, saturated brine, dried over anhydrous sodium sulfate, concentrated, column chromatography _{^{(eluent: CH 2 Cl 2: i PrOH}} = 60: 1 ~ 12: 1) to give the title Compound acetic acid (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)phosphonyl)oxy)methyl)-4-methyltetrahydrofuran 3-yl ester (52 mg, 90%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 8.92 (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

Preparation of the first step (2S)-2-((4-cyclopropylphenoxy)(perfluorophenoxy)phosphonium)amino)propylthiocarboxylic acid S-isopropyl ester

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-cyclopropyl phenyl dichlorophosphate.

Take S- isopropyl (S) -2- amino acid propyl 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 phenylphosphonium dichloride solution cooled at -78 ° C for about 2 hours, and the solution was slowly raised 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)-2-aminopropylthiocarboxylic acid S-isopropyl ester hydrochloride was added, and the mixture was dropwise added for 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) (EtOAcjjjjjj S-isopropyl ester of thiocarboxylic acid (1.07 g, 6%, epimer ratio: S _P /R _P = 2.5:1).

^{1 H NMR (400MHz, MeOD)} δ 7.2-6.4 (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)-2-(((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)phosphonium)amino)propylthiocarboxylic acid S- Preparation of isopropyl ester

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 (2S)-2-((4-cyclopropylphenoxy)(perfluorophenoxy)phosphonium)amino)propylthiocarboxylic acid was slowly added dropwise. THF S- isopropyl ester (1.0g, 1.9mmol) in (10 mL) added and the reaction was stirred overnight at this temperature, washed with 1N HCl solution (4mL) the reaction was quenched, concentrated the residue was added CH ₂ Cl ₂ (50mL) and 1N HCl solution (10 mL) was diluted, the organic phase was separated, and the aqueous phase was extracted with CH ₂ Cl ₂ (10 mL×2), and the organic phase was washed with saturated sodium hydrogen carbonate (10 mL) and brine (10 mL) Drying over sodium sulfate, column chromatography (eluent: CH ₂ Cl ₂ : ⁱ PrOH = 50:1 to 12:1) afforded the title compound (S)-2-(((S)-(4-cyclopropyl) Phenoxy)(((2R,3R,4R,5R)-5-(2,4-dicarbonyl-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy- 4-Methyltetrahydrofuran-2-yl)methoxy)phosphonium)amino)propyl thiocarboxylic acid S-isopropyl ester (0.28 g, 25%). ¹ H NMR (400 MHz, MeOD) δ 7.60 (d, J = 8.4 Hz, 1H), 7.16 (m, 2H), 7.08 (d, J = 8.4 Hz, 2H), 6.14 (d, J = 19.6 Hz, 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 (162MHz, MeOD) δ 3.68; ¹⁹ F NMR (376MHz, MeOD) δ -161.62; 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 diluted with the medium into a series of gradient concentrations during the experiment, which is 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 various concentrations of the test compound and the internal reference compound were added to the cultured cells 24 hours later. After 48 hours, luciferase activity was measured using a microplate reader. Compound half inhibitory concentration IC ₅₀ values can be by a series of different concentrations of the test compound to inhibit the enzymatic activity of luciferase values calculated.

2. Compound toxicity test on cells

The toxicity of the compounds of the invention to cells is determined by MTT cytotoxicity Assayed by the 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 diluted with the medium into a series of gradient concentrations during the experiment, which is 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 various concentrations of the test compound and the 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 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. Test data of the compound PK of the preferred embodiment of the present invention

HCV NS5B is an RNA polymerase responsible for HCV viral replication, and the substrate used for replication is a nucleoside triphosphate. The compound of the present invention is a prodrug of a phosphonium nucleoside nucleotide analog 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 analogs in vivo is directly related to the inhibitory activity against NS5B. Therefore, the inventors performed an in vitro PK test and an in vivo PK test to test the active drug of the preferred embodiment of the present invention. The ability of uridine triphosphate analogues, among which: 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: Prodrugs are metabolized in human hepatocytes to produce active drug triphosphate The pharmacokinetic test of fluorouridine was carried out using human liver cancer Huh-7 cells and human primary hepatocytes. 24 hours after the drug was added to the cell species colonization culture solution, a final concentration of 50 μ M. 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 next day by centrifugation (11,000 rpm, 10 minutes) to obtain a supernatant, of each supernatant sample were 100 μ L, 100 μ L of water was added, vortexed for 2 minutes, centrifugation (3500 rpm, 5 minutes) taken 10 The μL supernatant was subjected to LC/MS/MS measurement to analyze the concentration of uridine triphosphate.

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, and the mobile phase used (A) solution was 10 mM ammonium acetate (pH = 7.8), and (B) solution was Acetonitrile, flow rate 0.56mL/min, elution time 0-12.1 minutes, 0.01% 95% (A) and 5% (B), 4.2 minutes 100% (A), 4.5 minutes 20% (A) and 80% (B), 20% (A) and 80% (B) in 6 minutes, 95% (A) and 5% (B) in 6.1 minutes, 95% (A) and 5% (B) in 12 minutes , terminated in 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.

The compounds of the preferred embodiments 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:

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 triphosphate. Glycoside analogues.

In order to further demonstrate that the compounds of the preferred embodiments of the invention have the ability to metabolize the active drug triphosphate uridine analog, the inventors have also performed in vivo The PK assay was used to test the ability of the compound of the preferred embodiment of the invention to produce the active drug uridine analog. The in vivo PK model was: 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 by 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. The solid phase extraction was carried out using a weak anion exchange column from Waters, Inc., starting at 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. at 37 [deg.] C after nitrogen drying is redissolved in 160 μ L 5mM ammonium acetate, followed by addition of 40 μ L alkaline phosphatase treatment at 37 ℃ 30 minutes to give nucleoside 20 μ L sample after treatment for LC / MS / MS analysis of nuclear The concentration of glycosides.

The instrument used for LC/MS/MS analysis of nucleosides was AB Sciex API 5500 Qtrap. The conditions for liquid chromatography were Shimadzu LC-20AD pump, Luna 5 μm C18 30 x 2.0 mm column, (A) solution for mobile phase was 5 mM ammonium acetate, and (B) solution was 5 mM ammonium acetate / 95% acetonitrile / 5%. Water, flow rate 0.5 mL/min, elution time 0-4.5 minutes, 0.01% for 100% (A), 0.5 minutes for 100% (A), 1.2 minutes for 5% (A) and 95% (B), 2.4 minutes is 5% (A) and 95% (B), 2.5 minutes is 100% (A), and 4.5 minutes is 100% (A). The mass spectrometer was set up in the negative ion electrospray ionization (ESI) mode, the multi-reaction monitoring of the analyte fluorouridine was 259.20/239.20, and the multi-reaction monitoring of the internal reference AZT was 266.10/223.10. The main parameters of pharmacokinetics were calculated using WinNonlin 6.1, and the measurement data of the analyte fluorouridine was standardized 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. liver The sampling point of the 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. The solid phase extraction was carried out using a weak anion exchange column from Waters, Inc., starting at 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. redissolved with nitrogen at 37 [deg.] C with 160μL 5mM ammonium acetate and dried, followed by addition of 40 μ L alkaline phosphatase treatment at 37 ℃ 30 minutes to give nucleoside 20 μ L sample after treatment for LC / MS / MS analysis nucleosides concentration.

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

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

As can be seen from the data in Table 4, the compounds of the preferred embodiments of the present invention, such as the twenty-four, twenty-eight, thirty-two compounds, are also effectively metabolized in vivo to produce the active drug uridine uridine analog, similar to Sofosbuvir. In particular, the ability of the compound of the thirty-two compound to produce uridine triphosphate analogues in rat liver and canine liver was improved by more than 20% compared with the positive drug Sofosbuvir, and the twenty-four compounds in the example were in dogs. The ability to produce uridine triphosphate analogs in the liver is up to 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 solution is modified or equivalently substituted. For example, the first to the twenty-third embodiments 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 not to be construed as falling within the spirit and scope of the present invention.

Claims (33)

A uracil nucleotide analog of the compound of the following 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 ethoxylated; R _{1 is} selected from the group consisting of hydrogen, halogen, hydroxy, thiol, cyano, nitro, azide, C _1-8 alkyl, halogen 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 a carbon atom adjacent to the benzene ring of R ₁ forms 5 a -7 membered carbocyclic ring, a 5-7 membered heterocyclic ring wherein the C _1-8 alkyl group, C _2-8 alkenyl group, C _2-8 alkynyl group, C _3-8 cycloalkyl group, 3-8 member a heterocyclic group, a C _5-10 aryl group, a 5-10 membered heteroaryl group, a 5-7 membered carbocyclic ring or a 5-7 membered heterocyclic ring, further optionally one or more selected from the group consisting of halogen, hydroxy, thiol, cyano, Nitro, azido, 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 Substituted; 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} -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 )-C(O)OR ₅ the substituents; wherein the C _1-8 alkyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, C _5-10 aryl or 5-10 Heteroaryl optionally further substituted by one or more substituents selected from halo, hydroxy, mercapto, cyano, nitro, azido, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl group, 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 aryl oxygen , 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 Substituted by a substituent of OR ₅ ; R _{3 is} selected from hydrogen, C _1-8 alkyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, C _5-10 aryl or 5-10 member An aryl group, further optionally 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 group, 3-8 membered heterocyclic oxy group, 3-8 membered heterocyclic thio group, C _5-10 aryl group, C _5-10 aryl oxygen group, a C _5-10 aryl group, a 5-10 membered heteroaryl, 5-10 membered heteroaryl group, 5-10-membered heteroaryl _{Group, -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) Substituted by a substituent of R ₅ or -N(R ₅ )-C(O)OR ₅ ; R _{4 is} selected from the group consisting of hydrogen, halogen, hydroxy, thiol, cyano, nitro, azide, C _1-8 alkyl , halogen substituted C _1-8 alkyl, C _3-8 cycloalkyl, C _3-8 cycloalkylmethyl, halogen substituted C _1-8 alkoxy, halogen substituted C _1-8 alkylthio, 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 aryl thio 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 , -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 , R _{7 is} selected from the group consisting of hydrogen, C _1-4 alkyl, C _3-8 cycloalkyl; m is 0, 1, 2, 3, 4; r is 0, 1, 2. The uracil nucleotide analog, the stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 1, 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 the first item of the patent application. A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 2, wherein R _{1 is} selected from C _1-8 alkyl, halogen substituted C _1-8 Alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 membered heterocyclic, 3-8 membered heterocyclooxy, 3-8 member Cyclothio, C _5-10 aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy, 5- 10 member 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 , wherein the C _1-8 alkyl group, C _2-8 alkene a group, a C _2-8 alkynyl group, a C _3-8 cycloalkyl group, a 3-8 membered heterocyclic group, a C _5-10 aryl group or a 5-10 membered heteroaryl group, further optionally selected from one or more Halogen, hydroxy, decyl, cyano, nitro, azide, C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _3-8 cycloalkyl, 3-8 Heterocyclyl, 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 )- The substituent of C(O)OR ₅ is substituted; Y, R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , m, r are as defined in the first item of the patent application. The uracil nucleotide analog, the stereoisomer thereof or a pharmaceutically acceptable salt thereof, according to claim 3, wherein R _{1 is} selected from C _1-8 alkyl, halogen substituted C _1-8 An alkyl group, 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 chain The alkynyl group is further selected from 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 Substitution of aryl, C _5-10 aryloxy, C _5-10 arylthio, 5-10 membered heteroaryl, 5-10 membered heteroaryloxy or 5-10 membered heteroarylthio Substituted; Y, R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , m, r are as defined in the first item of the patent application. A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 4, which is selected from the group consisting of: A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 5, which is selected from the group consisting of: The uracil nucleotide analog, the stereoisomer thereof or a pharmaceutically acceptable salt thereof, according to the third aspect of the invention, wherein R _{1 is} selected from the group consisting of C _3-8 cycloalkyl, 3-8 Cyclo, 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, 3-8 membered heterocyclyl, C _5-10 aryl 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 group, 3-8 membered heterocyclic oxy group, 3-8 membered heterocyclic thio group, C _5-10 aryl group, 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 ₃ , R ₄ , R ₅ , R ₆ , R ₇ , m, r are as defined in item 1 of the scope of application. A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 7, which is selected from the group consisting of: A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 8, which is selected from the group consisting of: The uracil nucleotide analog, the stereoisomer thereof or a pharmaceutically acceptable salt thereof, according to claim 3, wherein 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 in the first item of the patent application. A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 10, which is selected from the group consisting of: A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 11, which is selected from the group consisting of: The uracil nucleotide analog, the stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 2, wherein the carbon atom adjacent to the benzene ring of R ₁ forms a 5-7 member carbon ring. , 5-7 membered heterocyclic ring, wherein the 5-7 membered carbocyclic or 5-7 membered heteroaryl surveying requires further by 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 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( Substituted by a substituent of R ₅ )-C(O)R ₅ or -N(R ₅ )-C(O)OR ₅ ; Y, R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , m, r As defined in item 1 of the patent application. The uracil nucleotide analog, the stereoisomer thereof or a pharmaceutically acceptable salt thereof, according to claim 13, wherein the carbon atom adjacent to the benzene ring of R ₁ forms a 5-7 member carbon ring. The 5-7 member heterocyclic ring is selected from the following structures: Wherein the 5-7 membered carbocyclic or 5-7 membered heteroaryl surveying requires further by 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 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 heteroarylsulfide Base, -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) Substituting for 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 set forth in claim 1 definition. The patentable scope of the application as item 14 uracil nucleotide analogues, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R ₁ to form a 5-7 membered carbocyclic ring carbon atom adjacent the benzene ring and The 5-7 member heterocyclic ring is selected from the following structures: The uracil nucleotide analog, the stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 1, 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, r are as defined in the first item of the patent application. The uracil nucleotide analog, the stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 16, wherein 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 , R ₃ , R ₅ , R ₆ , R ₇ , m, r are as defined in the first item of the patent application. The uracil nucleotide analog, the stereoisomer thereof or a pharmaceutically acceptable salt thereof, according to claim 17, wherein R _{3 is} selected from the group consisting of hydrogen, C _1-8 alkyl, C _3-8 a cycloalkyl group, a 3-8 membered heterocyclic group, a C _5-10 aryl group or a 5-10 membered heteroaryl group, further optionally one or more selected from the group consisting of halogen, hydroxy, thiol, 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 ,- _{Substituents for} C _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 Substituted; 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 1 item is defined. The uracil nucleotide analog, the stereoisomer thereof or a pharmaceutically acceptable salt thereof, according to claim 18, wherein R _{3 is} selected from the group consisting of hydrogen, C _1-4 alkyl, cyclopropyl, Cyclohexyl or phenyl, further optionally 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 - Substituted by a substituent of C(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 the first item of the patent application. The uracil nucleotide analog, the stereoisomer thereof or a pharmaceutically acceptable salt thereof, according to claim 19, wherein R _{1 is} selected from the group consisting of hydrogen, C _1-8 alkyl, halogen substituted C _{1 -8} alkyl, C _2-8 alkenyl, C _2-8 alkynyl, wherein the C _1-8 alkyl, halogen 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 heteroarylthio Substituted by a substituent; Y, R ₂ , R ₅ , R ₆ , R ₇ , m, r are as defined in the first item of the patent application. A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 20, which is selected from the group consisting of: The uracil nucleotide analog, the stereoisomer thereof or a pharmaceutically acceptable salt thereof, according to claim 19, wherein R _{1 is} selected from the group consisting of C _3-8 cycloalkyl and 3-8 Cyclo, 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, 3-8 membered heterocyclyl, C _5-10 aryl 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 group, 3-8 membered heterocyclic oxy group, 3-8 membered heterocyclic thio group, C _5-10 aryl group, 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 as defined in the first paragraph of the patent application scope. A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 22, which is selected from the group consisting of: The uracil nucleotide analog, the stereoisomer thereof or a pharmaceutically acceptable salt thereof, according to claim 19, 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 the first item of the patent application. A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 24, which is selected from the group consisting of: The patentable scope of the application as item 19 uracil nucleotide analogues, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R ₁ to form a 5-7 membered carbocyclic ring carbon atom adjacent the benzene ring and a 5-7 membered heterocyclic ring, wherein said 5-7 membered carbocyclic or 5-7 membered heterocyclic ring is further optionally 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 Base, 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 by a substituent of N(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 The scope of patent application is defined in item 1. The uracil nucleotide analog, the stereoisomer thereof or a pharmaceutically acceptable salt thereof, according to claim 26, wherein the carbon atom adjacent to the benzene ring of R ₁ forms a 5-7 member carbon ring. The 5-7 member heterocyclic ring is selected from the following structures: Wherein the 5-7 membered carbocyclic or 5-7 membered heteroaryl surveying requires further by 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 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 heteroarylsulfide Base, -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) Substituents of R ₅ or -N(R ₅ )-C(O)OR ₅ are substituted; Y, R ₂ , R ₅ , R ₆ , R ₇ , m, r are as defined for the compound of formula (I). A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 27, which is selected from the group consisting of: The uracil nucleotide analog, the stereoisomer thereof or a pharmaceutically acceptable salt thereof, according to any one of claims 16 to 28, wherein the stereoisomer is in the S configuration, the structure as follows: A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to claim 29, which is selected from the group consisting of: A method for preparing a uracil nucleotide analog, a stereoisomer thereof or a pharmaceutically acceptable salt thereof according to claim 1, wherein the method comprises the following steps: When Y is selected from the group consisting of ethyl hydrazino, the reaction further includes the following reaction as needed: Further, if necessary, column chromatography is carried out to obtain a stereoisomer thereof, or a stereoisomer thereof is obtained by the following steps: When Y is selected from the group consisting of ethyl hydrazino, the reaction further includes the following reaction as needed: Wherein: Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , m, r are as defined in the first item of the patent application. A pharmaceutical composition comprising a therapeutically effective amount of a uracil nucleotide analog according to item 1 of the patent application, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. A uracil nucleotide analog, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as described in claim 1, or a pharmaceutical composition according to claim 32, for use in preparation For the treatment of diseases caused by hepatitis C virus, hepatitis A virus, Sinan virus, yellow fever virus, dengue virus, rhinovirus, poliovirus, bovine viral diarrhea virus or Japanese encephalitis virus infection Applications.