Pyrimidine nucleoside peptide agricultural antibiotic and synthesis and application thereof

Abstract

The invention relates to the technical field of organic chemistry and medicinal chemistry, in particular to pyrimidine nucleoside peptide agricultural antibiotics and synthesis and application thereof. The specific technical scheme is as follows: the synthetic method of the pyrimidine nucleoside peptide compound comprises the following steps: stirring a pyrimidine amino saccharide compound and a dipeptide compound for 0-96 hours under the action of an organic solvent, an HATU condensing agent and alkali, and separating and purifying to obtain the fully-protected pyrimidine nucleoside peptide compound, and then carrying out a two-step reaction to remove a sugar ring protecting group and a dipeptide branched chain protecting group so as to obtain the pyrimidine nucleoside peptide compound. The invention solves the problems of limited types of pyrimidine nucleoside peptide compounds obtained by microbial fermentation, low yield, long route, serious pollution and the like when the compounds are obtained by chemical synthesis in the prior art.

Description

Technical Field

[0001] The invention relates to the technical field of organic chemistry and medicinal chemistry, and in particular to a pyrimidine nucleoside peptide agricultural antibiotic and the synthesis and application thereof. Background Art

[0002] Biopesticides have been rapidly developed in recent decades due to their green, environmentally friendly, safe and efficient characteristics (Journal of Pesticide Science, 2023, 25, 769-778). Among them, nucleoside antibiotics have aroused great interest among chemical synthesis workers due to their wide range of biological activities (J. Antibiot. 1988, 41, 1711; Pharm. Therap. 1991, 52, 269; Nat. Prod. Rep. 2010, 27, 279; Trends in Microbiology 2015, 23, 110; Org. Biomol. Chem. 2021, 19, 4285, etc.). Many literatures have reported the good application prospects of some nucleoside antibiotics in crop protection (Chem. Rev. 1992, 92, 1745; Chem. Rev. 1995, 95, 1859; Heterocycles 2005, 65, 667; Pest Manag. Sci. 2007, 63, 524; Progress in Chemistry, 2019, 31, 954, etc.).

[0003] Most of the nucleoside compounds isolated from microbial secondary metabolites have been developed into agricultural antibiotics. They are easy to degrade, have little environmental pollution, and have low toxicity to animals. They are an ideal green biological pesticide and play an important role in the prevention and control of crop diseases and insect pests. As early as 1958, Japanese scientists isolated Blasticidin S from the fermentation broth of Streptomyces griseus and soon developed it into a biological pesticide, which was widely used in Japan to prevent and control rice blast (Tetrahedron Lett. 1965, 6, 1411). Subsequently, Japanese scientists isolated Gougerotin from Streptomyces graminis and Streptomyces graminis. It has a broad spectrum of biological activities such as antibacterial, anthelmintic and acaricidal, and is a very valuable agricultural antibiotic. It is also the most studied nucleoside agricultural antibiotic (J. Antibiot. 1962, 15, 93). In addition, Yunnanmycin, extracted and separated from the fermentation broth of Streptomyces strains, has the effect of inhibiting the growth of rice pathogenic fungi and tobacco pathogenic fungi (Chinese J. Antibiot. 1998, 16, 3). Ningnanmycin is a nucleoside agricultural antibiotic discovered by the Chengdu Institute of Biology, Chinese Academy of Sciences from the fermentation broth of the Xichang variant of Streptomyces noursei (Acta Microbiologica Sinica, 1995, 35, 368-374). Neomycin is another nucleoside agricultural antibiotic isolated from the fermentation broth of Streptomyces noursei by researchers at the Chengdu Institute of Biology (Pesticides, 2013, 52, 293), which has a broad spectrum of biological activities such as inhibiting bacteria, plant fungi and antiviral.

[0004] However, the yield of nucleoside agricultural antibiotics obtained by microbial fermentation is low and the structural diversity is limited, resulting in a single product and limited new product development. Therefore, it is of great significance to obtain high-purity nucleoside agricultural antibiotics by chemical total synthesis. However, the current method for chemically synthesizing such agricultural nucleoside antibiotics has a long synthesis route, low yield, and complex post-processing (Org. Biomol. Chem. 2021, 19, 4285, CN107973830B), which is difficult to meet the needs of the development of green pesticides. Summary of the invention

[0005] In view of the shortcomings of the prior art, the present invention provides a pyrimidine nucleoside peptide agricultural antibiotic and its synthesis and application, which solves the problems in the prior art of limited types of pyrimidine nucleoside peptide compounds obtained from microbial fermentation, low yield, long route and high pollution when obtaining such compounds by chemical synthesis.

[0006] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0007] The present invention discloses a pyrimidine nucleoside peptide agricultural antibiotic, the structural formula of which is any one of the following formulas (I):

[0008]

[0009] In the formula, R 1 is any one of a hydrogen atom, an alkyl group, an aryl group, a heteroaryl group, an alkoxy group, an amino group, a substituted amino group, an ester group, an alkenyl group, an alkynyl group, a carboxyl group, a halogen group, a nitro group, a fluoroalkyl group, a cyano group, a sulfone group, an amide group, and a phosphate group;

[0010] R 2 It is any one of an alkoxy group, a hydroxyl group, an amino group, a substituted amine group, and a cyclic amine group;

[0011] R 3 , R 4 are any one of hydrogen atom, aromatic acyl group, heteroaromatic acyl group, fatty acyl group and sulfonyl group; or, R 3 O.R 4 O is substituted by any one of an azide group, a substituted amino group, a thiol group, an alkyl sulfide, an aryl sulfide, an alkyl group, an alkenyl group, and an alkynyl group;

[0012] R 5 is any one of a hydroxyl group, an ester group, an alkoxy group, an alkyl group, an aryl group, a heteroaryl group, an azido group, an amino group, a substituted amino group, a dialkylamino group, and a cyclic amino group;

[0013] R 6 is any one of a hydrogen atom, a benzyloxycarbonyl group, a tert-butyloxycarbonyl group, an alkyl group, an aromatic acyl group, a fatty acyl group, and a sulfonyl group;

[0014] n is any natural number from 0 to 10.

[0015] Preferably, when the R 1 When R is any one of a hydrogen atom, an alkyl group, an aryl group, a heteroaryl group, an alkoxy group, an amino group, a substituted amino group, an ester group, an alkenyl group, an alkynyl group, a carboxyl group, a halogen group, a nitro group, a fluoroalkyl group, a cyano group, a sulfone group, an amide group, and a phosphate group, 1 Can be represented by one or more independent R X Substituents are substituted;

[0016] When the R 2 When R is any one of an alkoxy group, a hydroxyl group, an amino group, a substituted amine group, and a cyclic amine group; 2 Can be represented by one or more independent R X Substituents are substituted;

[0017] When the R 3 , R 4 When R3 , R 4 Can be represented by one or more independent R X Substituents are substituted;

[0018] Or, when the R 3 O.R 4 When O is independently any one of an azido group, a substituted amino group, a mercapto group, an alkyl sulfide, an aryl sulfide, an alkyl group, an alkenyl group, and an alkynyl group, R 3 O.R 4 O can be represented by one or more independent R X Substituents are substituted;

[0019] When the R 5 When R is any one of a hydroxyl group, an ester group, an alkoxy group, an alkyl group, an aryl group, a heteroaryl group, an azido group, an amino group, a substituted amino group, a dialkylamino group, and a cyclic amino group, 5 Can be represented by one or more independent R X Substituents are substituted;

[0020] When the R 6 When R is any one of a hydrogen atom, a benzyloxycarbonyl group, a tert-butyloxycarbonyl group, an alkyl group, an aromatic acyl group, an aliphatic acyl group, and a sulfonyl group, 6 Can be represented by one or more independent R X Substituents are substituted;

[0021] The R X It is any one of a hydrogen atom, a halogen, an alkyl group, a fluoroalkyl group, an alkenyl group, an alkynyl group, a nitro group, a cyano group, a cycloalkyl group, an aryl group, an amino group, an alkoxy group, a substituted amino group, an amide group, a hydroxyl group, a sulfonamide group, a diazo group, an azido group, an alkyl acyl group, and an aryl acyl group.

[0022] Preferably, when the R X When R is any one of a hydrogen atom, a halogen, an alkyl group, a fluoroalkyl group, an alkenyl group, an alkynyl group, a nitro group, a cyano group, a cycloalkyl group, an aryl group, an amino group, an alkoxy group, a substituted amino group, an amide group, a hydroxyl group, a sulfonamide group, a diazo group, an azido group, an alkyl acyl group, and an aryl acyl group, X It may be substituted by one or more hydrogen atoms, halogen, alkyl, fluoroalkyl, alkenyl, alkynyl, nitro, cyano, cycloalkyl, aryl, amino, alkoxy, substituted amino, amide, hydroxyl, sulfonamide, diazo, azido, alkyl acyl, aryl acyl, respectively.

[0023] Accordingly, a method for synthesizing a pyrimidine nucleoside peptide agricultural antibiotic comprises adding compound 7 or compound 8 and Pd / C to a mixed solvent of EtOH and water, reacting completely at room temperature in the presence of hydrogen, filtering to remove Pd / C, and concentrating to obtain a pyrimidine nucleoside peptide compound;

[0024] The compound 7 or compound 8 is synthesized by compound 6, and the synthesis process is as follows: dissolving compound 6 in CH3OH, adding an aqueous solution of LiOH at 0°C for hydrolysis, the reaction is complete, adjusting the pH to 2 with glacial acetic acid, retaining the aqueous phase, concentrating and filtering the aqueous phase, and separating and purifying the filtrate by HPLC to obtain compound 7 or compound 8; the equivalent ratio of LiOH to compound 6 is 0.01:1 to 50:1; the structural formula of compound 6 is:

[0025]

[0026] Preferably, the synthesis method of compound 6 is: using compound 4 and compound 5 as raw materials, HATU as a condensation agent, 2,6-dimethylpyridine as a base, fully reacting under argon protection conditions, extracting, washing, drying, filtering and concentrating, and separating by silica gel column chromatography to obtain compound 6;

[0027] The equivalent ratio of compound 5 to compound 4 is 0.01:1 to 50:1, the equivalent ratio of HATU to compound 4 is 0.01:1 to 50:1, and the equivalent ratio of 2,6-lutidine to compound 4 is 0.01:1 to 50:1;

[0028] The structural formulas of the compounds 4 and 5 are:

[0029]

[0030] Preferably, the synthesis process of compound 4 is:

[0031] 1) Compound 3 is dissolved in a mixed solvent of CH3CN and H2O, TEMPO is added, and then iodobenzene acetate is added. After the reaction is complete, anhydrous CH3OH is added to quench the reaction, and the crude carboxylic acid compound is concentrated to obtain a crude carboxylic acid compound, which can be subjected to the next step of reaction after pulping; the equivalent ratio of TEMPO to compound 3 is 0.001:1 to 10:1, and the equivalent ratio of iodobenzene acetate to compound 3 is 0.001:1 to 100:1;

[0032] 2) the carboxylic acid compound is dissolved in CH3OH, H2SO4 is added, refluxed, concentrated after the reaction is complete, the reaction solution is extracted with ethyl acetate, the organic phase is retained, dried, filtered, and concentrated under reduced pressure to obtain a crude methyl esterification product; the equivalent ratio of H2SO4 to the carboxylic acid compound is 0.001:1 to 50:1;

[0033] 3) reacting the methyl ester compound with SnCl2·2H2O, p-Toluenethiol, CHCl3 and Et3N until the reaction is complete, concentrating under reduced pressure, adding ethyl acetate to dissolve, filtering to remove the insoluble SnCl2·2H2O, concentrating the filtrate, and separating and purifying by silica gel column chromatography to obtain compound 4;

[0034] The equivalent ratio of SnCl2·2H2O to the methyl ester compound is 0.01:1 to 50:1, the equivalent ratio of p-methylthiophenol to the methyl ester compound is 0.01:1 to 100:1, and the equivalent ratio of Et3N to the methyl ester compound is 0.01:1 to 300:1;

[0035] The structural formula of the compound 3 is:

[0036]

[0037] Preferably, the synthesis process of compound 3 is: using 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative as substrate, adding uracil or uracil with different substituents, adding catalyst, BSA and acetonitrile, and carrying out condensation reaction under heating reflux conditions to obtain nucleoside products;

[0038] The catalyst is any one of BF3·Et2O, TMSOTf, TBSOTf, TESOTf, metal halide, metal sulfonate, perchloric acid and its metal salt, triarylborane, organic phosphoric acid, organic thiourea, and organic carbonyl acid.

[0039] Preferably, the equivalent ratio of the catalyst to the 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative is 1:0.01 to 1:100, the equivalent ratio of uracil or substituted uracil to the 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative is 1:0.01 to 1:100, the equivalent ratio of BSA to the 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative is 1:0.01 to 1:100, the equivalent ratio of the nucleoside product to sodium hydroxide is 1:0.01 to 1:100, the reaction concentration is 0.01 to 100 mol / L, and the reaction temperature is 60 to 200°C.

[0040] Correspondingly, an agricultural antibiotic composition comprises the pyrimidine nucleoside peptide agricultural antibiotic or the pyrimidine nucleoside peptide agricultural antibiotic synthesized by the synthesis method, or its stereoisomers, tautomers or salts, or its prodrug molecules and a pesticide acceptable carrier.

[0041] Correspondingly, the use of the pyrimidine nucleoside peptide agricultural antibiotic or the pyrimidine nucleoside peptide agricultural antibiotic synthesized by the synthesis method, or its stereoisomers, tautomers or salts, or its prodrug molecules, or the agricultural antibiotic composition in the prevention and control of plant diseases, wherein the plant diseases include rapeseed sclerotinia rot, rice blast, and wheat fusarium rust.

[0042] The present invention has the following beneficial effects:

[0043] 1. The present invention uses 1-O-acetyl-4-azido-4-deoxy-D-glucose derivatives as starting materials and realizes the chemical total synthesis and activity evaluation of a pyrimidine nucleoside peptide agricultural antibiotic through a series of reactions. The invention has the characteristics of mild reaction conditions, low cost, simple operation and high product purity.

[0044] 2. The present invention synthesizes a series of pyrimidine nucleoside peptide agricultural antibiotics, which provide a material basis for the discovery of new biological pesticide molecules with high efficiency, low toxicity and low residue. At the same time, it exhibits a wide range of anti-plant disease activities. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 is the inhibition rate of compounds W1~W12 on rapeseed sclerotinia. DETAILED DESCRIPTION

[0046] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0047] Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art.

[0048] 1. The present invention discloses a pyrimidine nucleoside peptide agricultural antibiotic, the structural formula of which is any one of the following formula (I):

[0049]

[0050] In the formula, R 1 is any one of a hydrogen atom, an alkyl group, an aryl group, a heteroaryl group, an alkoxy group, an amino group, a substituted amino group, an ester group, an alkenyl group, an alkynyl group, a carboxyl group, a halogen group, a nitro group, a fluoroalkyl group, a cyano group, a sulfone group, an amide group, and a phosphate group;

[0051] R 2 It is any one of an alkoxy group, a hydroxyl group, an amino group, a substituted amine group, and a cyclic amine group;

[0052] R3 , R 4 are any one of hydrogen atom, aromatic acyl group, heteroaromatic acyl group, fatty acyl group and sulfonyl group; or, R 3 O.R 4 O is substituted by any one of an azide group, a substituted amino group, a thiol group, an alkyl sulfide, an aryl sulfide, an alkyl group, an alkenyl group, and an alkynyl group;

[0053] R 5 is any one of a hydroxyl group, an ester group, an alkoxy group, an alkyl group, an aryl group, a heteroaryl group, an azido group, an amino group, a substituted amino group, a dialkylamino group, and a cyclic amino group;

[0054] R 6 is any one of a hydrogen atom, a benzyloxycarbonyl group, a tert-butyloxycarbonyl group, an alkyl group, an aromatic acyl group, a fatty acyl group, and a sulfonyl group;

[0055] n is any natural number from 0 to 10.

[0056] Furthermore, when the R 1 When R is any one of a hydrogen atom, an alkyl group, an aryl group, a heteroaryl group, an alkoxy group, an amino group, a substituted amino group, an ester group, an alkenyl group, an alkynyl group, a carboxyl group, a halogen group, a nitro group, a fluoroalkyl group, a cyano group, a sulfone group, an amide group, and a phosphate group, 1 Can be represented by one or more independent R X Substituents are substituted;

[0057] When the R 2 When R is any one of an alkoxy group, a hydroxyl group, an amino group, a substituted amine group, and a cyclic amine group; 2 Can be represented by one or more independent R X Substituents are substituted;

[0058] When the R 3 , R 4 When R 3 , R 4 Can be represented by one or more independent R X Substituents are substituted;

[0059] Or, when the R 3 O.R 4 When O is independently any one of an azido group, a substituted amino group, a mercapto group, an alkyl sulfide, an aryl sulfide, an alkyl group, an alkenyl group, and an alkynyl group, R 3 O.R 4 O can be represented by one or more independent R X Substituents are substituted;

[0060] When the R 5When R is any one of a hydroxyl group, an ester group, an alkoxy group, an alkyl group, an aryl group, a heteroaryl group, an azido group, an amino group, a substituted amino group, a dialkylamino group, and a cyclic amino group, 5 Can be represented by one or more independent R X Substituents are substituted;

[0061] When the R 6 When R is any one of a hydrogen atom, a benzyloxycarbonyl group, a tert-butyloxycarbonyl group, an alkyl group, an aromatic acyl group, an aliphatic acyl group, and a sulfonyl group, 6 Can be represented by one or more independent R X Substituents are substituted;

[0062] The R X It is any one of a hydrogen atom, a halogen, an alkyl group, a fluoroalkyl group, an alkenyl group, an alkynyl group, a nitro group, a cyano group, a cycloalkyl group, an aryl group, an amino group, an alkoxy group, a substituted amino group, an amide group, a hydroxyl group, a sulfonamide group, a diazo group, an azido group, an alkyl acyl group, and an aryl acyl group.

[0063] Furthermore, when the R X When R is any one of a hydrogen atom, a halogen, an alkyl group, a fluoroalkyl group, an alkenyl group, an alkynyl group, a nitro group, a cyano group, a cycloalkyl group, an aryl group, an amino group, an alkoxy group, a substituted amino group, an amide group, a hydroxyl group, a sulfonamide group, a diazo group, an azido group, an alkyl acyl group, and an aryl acyl group, X It may be substituted by one or more hydrogen atoms, halogen, alkyl, fluoroalkyl, alkenyl, alkynyl, nitro, cyano, cycloalkyl, aryl, amino, alkoxy, substituted amino, amide, hydroxyl, sulfonamide, diazo, azido, alkyl acyl, aryl acyl, respectively.

[0064] 2. The present invention discloses a method for synthesizing a pyrimidine nucleoside peptide agricultural antibiotic, using a 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative as a raw material, and preparing a pyrimidine nucleoside peptide agricultural antibiotic compound through condensation, hydrolysis, oxidation, esterification, reduction and other reactions in an organic solvent and / or water solvent. Specifically, compound 6 is obtained through 6 steps of reaction, and then the pyrimidine nucleoside peptide compound is obtained through 2 steps of sequential deprotection.

[0065] As one specific synthetic route, a 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative is selected as (1-O-acetyl-2,3,6-tri-O-benzoyl-4-azido-4-deoxy-D-glucose (compound 1); compound 6 is selectively deprotected by removing the protecting group R 2 , R 3 , R 4 To prepare compound 7 or compound 8, specifically compound 6 is R 2 =OMe, R 3 =Bz(benzoyl), R4 =Bz (benzoyl) as an example, compound 4 is R 2 =OMe, R 3 =Bz(benzoyl), R 4 =Bz (benzoyl) as an example; Compound 3 is R 3 =Bz(benzoyl), R 4 =Bz (benzoyl) as an example. The specific synthesis route is as follows:

[0066]

[0067] The reaction concentration in all the above reaction steps is 0.001 to 50 mol / L, the reaction temperature is -100 to 200°C, and the reaction time of each step in all the steps is 0.1 to 96 hours. The reaction concentration here refers to the amount of the substance of the reaction substrate in a unit volume of solvent (i.e., the amount of the substance of the reaction substrate in 1L of solvent is 0.001 to 50 mol).

[0068] A catalyst needs to be added to the reaction system for synthesizing compound 3 from compound 1, and the equivalent ratio of the catalyst to compound 1 is 1:0.01 to 1:100. The catalyst is any one of BF3·Et2O, TMSOTf, TBSOTf, TESOTf, metal halide, metal sulfonate, perchloric acid and its metal salt, triarylborone, organic phosphoric acid, organic thiourea, and organic carbonyl acid.

[0069] The organic solvent used in the whole reaction process is any one of 2,2,2-trifluoroethanol, 2,2,2-trichloroethanol, 1,1,1,3,3,3-hexafluoro-2-2-propanol, chloroethanol, 2,2-dichloroethanol, dichloromethane, chloroform, acetonitrile, methanol, ethanol, n-butanol, tert-butanol, isopropanol, benzene, toluene, xylene, chlorobenzene, tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, 1,4-dioxane, 2-methyltetrahydrofuran, ether, and methyl tert-butyl ether, or a mixture of several solvents in any proportion.

[0070] The specific synthesis process is:

[0071] (1) The structural formula of compound 3 is:

[0072]

[0073] The synthesis process of compound 3 is as follows: using 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative as substrate, respectively adding uracil or uracil with different substitutions, adding catalyst, BSA and acetonitrile, and carrying out condensation reaction under heating reflux conditions to obtain nucleoside products; the nucleoside products are selectively hydrolyzed with 6-benzoyl protecting group under the action of NaOH and MeOH to obtain compound 3;

[0074] The catalyst is any one of BF3·Et2O, TMSOTf, TBSOTf, TESOTf, metal halide, metal sulfonate, perchloric acid and its metal salt, triarylborone, organic phosphoric acid, organic thiourea, and organic carbonyl acid;

[0075] The equivalent ratio of the catalyst to the 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative is 1:0.01-1:100, the equivalent ratio of uracil or substituted uracil to the 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative is 1:0.01-1:100, the equivalent ratio of BSA to the 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative is 1:0.01-1:100, the equivalent ratio of the nucleoside product to sodium hydroxide is 1:0.01-1:100, the reaction concentration is 0.01-100 mol / L, and the reaction temperature is 60-200°C.

[0076] (2) The structural formula of compound 4 is:

[0077]

[0078] The synthesis process of compound 4 is as follows:

[0079] 1) Compound 3 is dissolved in a mixed solvent of CH3CN and H2O, TEMPO is added, and then iodobenzene acetate is added to oxidize the primary hydroxyl group at position 6 of the sugar ring to a carboxyl group. After the reaction is complete, anhydrous CH3OH is added to quench the reaction, and the crude carboxylic acid compound is concentrated to obtain a crude product, which can be subjected to the next step of reaction after simple beating; the equivalent ratio of TEMPO to compound 3 is 0.001:1 to 10:1, and the equivalent ratio of iodobenzene acetate to compound 3 is 0.001:1 to 100:1;

[0080] 2) The carboxylic acid compound is dissolved in CH3OH, H2SO4 is added, and the mixture is refluxed to convert the 6-position carboxyl group into a methyl ester through an esterification reaction. After the reaction is complete, the mixture is concentrated, and the reaction solution is extracted with ethyl acetate. The organic phase is retained, dried, filtered, and concentrated under reduced pressure to obtain a crude methyl esterification product; the equivalent ratio of H2SO4 to the carboxylic acid compound is 0.001:1 to 50:1; It should be noted that step 1) is to first oxidize the hydroxymethyl group to a carboxylic acid (R 2=OH), step 2) is to form the corresponding methyl ester (R 2 =OMe), in this step, different alcohols or amines can be replaced to obtain different R 2 Substituted compound 4.

[0081] 3) reacting the methyl ester compound with SnCl2·2H2O, p-methylthiophenol, CHCl3 and Et3N, and concentrating under reduced pressure until the reaction is complete, adding ethyl acetate to dissolve, filtering to remove the insoluble SnCl2·2H2O, concentrating the filtrate, and separating and purifying by silica gel column chromatography to obtain compound 4;

[0082] The equivalent ratio of SnCl2·2H2O to the methyl ester compound is 0.01:1-50:1, the equivalent ratio of p-methylthiophenol to the methyl ester compound is 0.01:1-100:1, and the equivalent ratio of Et3N to the methyl ester compound is 0.01:1-300:1.

[0083] (3) The structural formula of compound 6 is:

[0084]

[0085] The synthesis process of compound 6 is: using compound 4 and compound 5 as raw materials, HATU as a condensation agent, 2,6-lutidine as a base, fully reacting under argon protection conditions, extracting, washing, drying, filtering and concentrating, and separating by silica gel column chromatography to obtain compound 6; the synthesis method of compound 5 can be found in the literature: Org.Biomol.Chem.2021,19,4285-4291, or patent: CN107973830B.

[0086] The equivalent ratio of compound 5 to compound 4 is 0.01:1 to 50:1, the equivalent ratio of HATU to compound 4 is 0.01:1 to 50:1, and the equivalent ratio of 2,6-lutidine to compound 4 is 0.01:1 to 50:1;

[0087] The structural formula of the compound 5 is:

[0088]

[0089] (4) The synthesis process of compound 7 or compound 8 is as follows: dissolve compound 6 in CH3OH, add an aqueous solution of LiOH at 0°C for hydrolysis, and after the reaction is complete, adjust the pH to 2 with glacial acetic acid, retain the aqueous phase, concentrate and filter the aqueous phase, and separate and purify the filtrate by HPLC to obtain compound 7 or compound 8; the equivalent ratio of LiOH to compound 6 is 0.01:1 to 50:1.

[0090] (5) The synthesis process of pyrimidine nucleoside peptide antibiotics is as follows: Compound 7 or Compound 8 and 20% Pd / C are added to a mixed solvent of EtOH and water, reacted completely at room temperature under hydrogen conditions, Pd / C was removed by filtration, and concentrated to obtain pyrimidine nucleoside peptide compounds, i.e., pyrimidine nucleoside peptide antibiotics.

[0091] 3. An agricultural antibiotic composition comprising a pyrimidine nucleoside peptide agricultural antibiotic, or a stereoisomer, a tautomer or a salt thereof, or a prodrug molecule thereof and an agrochemically acceptable carrier.

[0092] 4. Use of a pyrimidine nucleoside peptide agricultural antibiotic, or its stereoisomers, tautomers or salts, or its prodrug molecules, or an agricultural antibiotic composition in the prevention and treatment of plant diseases, wherein the plant diseases include rapeseed sclerotinia rot, rice blast, and wheat fusarium rust.

[0093] In order to further illustrate the synthesis of the pyrimidine nucleoside peptide compounds in the present invention, the present invention selectively synthesized the following substances, and illustrated the synthesis process of the following W1 to W12 compounds in conjunction with specific examples.

[0094] Since the present invention involves a series of chemical reactions, in order to enable those skilled in the art to understand the technical solution of the present invention, a detailed embodiment is provided: 3 , R 4 Both are benzoyl (Bz), R 2 is benzoyloxy (OBz) or hydroxy or methoxy, R 6 The benzyloxycarbonyl (Cbz) is taken as an example for detailed description. It should be clear to those skilled in the art that, based on this embodiment, appropriate adjustment of the synthetic raw materials, process methods and parameters to achieve the same purpose is within the scope of protection of the present invention.

[0095]

[0096] Example 1 Total Synthesis of W1 to W4

[0097] 1. Synthesize compound 3-1, the chemical reaction equation is:

[0098]

[0099] The specific synthesis process is:

[0100] Add 5-F substituted uracil (BS1) (1.74 g, 13.41 mmol) and BSA (5.92 mL, 35.76 mmol) to dry CH3CN (90 mL), stir at room temperature for 30 minutes, and the reaction solution changes from turbid to clear. Add compound 1 (5.0 g, 8.94 mmol), followed by TMSOTf (5.84 mL, 32.18 mmol). Under N2 protection, react at 77 ° C, and monitor the reaction by TLC. Concentrate, extract the reaction solution with water and ethyl acetate (3 × 200 mL). Combine the ethyl acetate layers, wash with saturated NaCl solution (2 × 200 mL), dry over anhydrous Na2SO4, filter, and concentrate under reduced pressure to obtain a crude nucleoside compound.

[0101] The crude nucleoside compound (5.63 g, 8.94 mmol) was dissolved in anhydrous CH3OH (40 mL), and NaOH (0.357 g, 8.94 mmol) was added. After reacting at room temperature for 1-3 hours, the reaction was complete as monitored by TLC, and the 6-position protecting group was selectively removed. 1M HCl was added to quench the reaction, and the concentrate was concentrated under reduced pressure. The obtained concentrate was separated by silica gel column chromatography (dichloromethane / methanol, 50:1) to obtain a white solid compound 3-1 (2.66 g, 57%, two steps).

[0102] The obtained compound 3-1 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0103] 1 H NMR (400MHz, CD3OD) δ8.23(dd,J=6.6,1.5Hz,1H),7.99(d,J=7.7Hz,2H),7.87(d,J=7.7Hz,2H),7.61–7.50(m,2H),7.40(dt,J=21.6,7.3Hz,4H ),6.17(d,J=9.1Hz,1H),5.96(td,J=9.8,1.5Hz,1H),5.64(t,J=9.3Hz,1H),4.22(t,J=9.9Hz,1H),3.96(d,J=12.2Hz,1H),3.92–3.82(m,2H). 13 C NMR(101MHz,CD3OD)δ165.51,165.35,157.75,157.49,149.33,141.83,139.50,133.51,133.45,12 9.44,129.33,128.76,128.30,128.22,125.26,124.91,80.70,77.53,73.91,70.96,60.40,59.18. 19F NMR(376MHz,CD3OD)δ-167.65.ESI-HRMS(m / z):calcd for C 24 H 20 FN5O8:[M+Na] + 548.11881; found:548.11811.

[0104] 2. Synthesize compound 4-1, the chemical reaction equation is:

[0105]

[0106] The specific synthesis process is:

[0107] Compound 3-1 (2.66 g, 5.06 mmol) was dissolved in CH3CN (15 mL) and water (12 mL), TEMPO (250 mg, 1.6 mmol) was added, and iodobenzene acetate (4.16 g, 15.63 mmol) was added in batches, and stirred at room temperature until the reaction of compound 3-1 was complete as monitored by TLC. CH3OH (5 mL) was added to quench the reaction, and the mixture was concentrated and briefly slurried with n-hexane to obtain a crude carboxylic acid product, which could be directly used for the next step.

[0108] The crude carboxylic acid product was dissolved in anhydrous CH3OH (40 mL), H2SO4 (1.01 mL) was added, and the reaction was carried out at room temperature. The reaction was completed after TLC monitoring. NaHCO3 was added to quench the reaction and concentrated. The reaction solution was extracted with water (40 mL) and ethyl acetate (3×100 mL), and the combined ethyl acetate solution was washed with saturated NaCl solution (2×50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain a crude product of the 6-position methyl ester of the sugar ring, which can be directly used for the next step reaction.

[0109] SnCl2·2H2O (2.28 g, 10.12 mmol) and p-methylthiophenol (2.51 g, 20.24 mmol) were added to CHCl3 (35 mL), and Et3N (4.22 mL, 30.36 mmol) was added dropwise at room temperature. Stir for 5 minutes, and the above crude methyl ester (2.80 g, 5.06 mmol) was added. The reaction was carried out at room temperature, and the reaction was monitored by TLC. The solvent was dried, ethyl acetate was added to dissolve, the insoluble matter SnCl2·2H2O was filtered out, dried, silica gel was added to mix the sample, and separated by silica gel column chromatography (CH2Cl2 / CH3OH, 40:1) to obtain white solid compound 4-1 (1.86 g, 70%, three steps).

[0110] The intermediate 4-1 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0111] 1 H NMR (400MHz, DMSO-d6) δ8.48(d,J=7.0Hz,1H),7.90(d,J=7.7Hz,2H),7.76(d,J=7.7Hz,2H),7.61(t,J=7.4Hz,3 H),7.46(dt,J=10.0,7.6Hz,5H),6.17(d,J=8.6Hz,1H),5.77–5.66(m,2H),4.50(d,J=10.1Hz,1H),3.73(s,3H). 13 C NMR (101MHz, DMSO-d6) δ168.67,165.97,165.15,157.33,157.06,149.55,141.81,134.36,133. 95,129.76,129.71,129.60,129.29,129.12,128.73,80.66,78.46,76.04,71.31,53.46,52.74. 19 F NMR(376MHz,DMSO-d6)δ-166.36.ESI-HRMS(m / z):calcd for C 25 H 22 FN3O9:[M+H] + 528.14128; found:528.14121.

[0112] 3. Synthesize compound W1, the chemical reaction equation is:

[0113]

[0114] The specific synthesis process is:

[0115] The above compound 5-1 (1.3 g, 4.18 mmol) was dissolved in dry DMF (30 mL), and then HATU (2.89 g, 7.60 mmol) and 2,6-dimethylpyridine (1.84 mL, 15.77 mmol) were added in sequence, and stirred at room temperature for 2 hours. Compound 4-1 (1.0 g, 1.90 mmol) was added, and stirring was continued at room temperature for 14 hours. After TLC monitoring, the reaction of the raw materials was complete. Saturated NaCl solution (50 mL) was added, and the reaction solution was extracted with ethyl acetate (3×100 mL). The combined ethyl acetate solution was washed with saturated NaCl solution (2×30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The obtained concentrate was separated by silica gel column chromatography (CH2Cl2 / CH3OH, 50:1) to obtain a light yellow solid compound W1 (1.04 g, 66%).

[0116] The obtained product W1 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0117] 1 H NMR (400MHz, CDCl3) δ7.87 (m, J=25.9, 8.2Hz, 3H), 7.72 (dd, J=10.0, 5.5Hz, 1H), 7.52 (t,J=7.5Hz,1H),7.43–7.30(m,8H),7.12(dd,J=18.0,10.2Hz,2H),6.33(t,J=10.7Hz ,1H),6.04(t,J=9.8Hz,1H),5.79–5.61(m,1H),5.18(d,J=11.9Hz,2H),4.69(s,1H), 4.03(s,1H),3.80(d,J=11.5Hz,3H),3.04(d,J=5.5Hz,2H),2.90(s,2H),1.81(s,3H). 13 C NMR (151MHz, CDCl3) δ171.04,170.56,167.31,166.31,165.25,156.49,149.34,141.87,136.15,133.79,129.82,128.58,128.46,128.24, 128.17,127.78,80.38,75.43,72.89,71.71,69.95,69.62,67.96,61 .44,54.10,53.29,52.12,51.51,36.11,31.93,31.62,31.44,29.70. 19 F NMR(376MHz, CDCl3)δ-161.76,-161.86.ESI-HRMS(m / z):calcdfor C 39 H 38 F5O 14 :[M+Na] + 842.22915;found:842.23080.[a] D 20 +13.2 (c 0.12, DMF).

[0118] 4. Synthesize compound W2, the chemical reaction equation is:

[0119]

[0120] The specific synthesis process is:

[0121] Compound 6-1 (500 mg, 0.61 mmol) was weighed in a bottle, and ammonia methanol solution (7 mol / L) (5 mL) was added under argon protection, and stirred at room temperature until the reaction of the raw material was complete as monitored by TLC. The concentrate was spin-dried and separated by silica gel column chromatography (CH2Cl2 / CH3OH, 40:1) to obtain a light yellow solid compound W2 (55.3 mg, 11%).

[0122] The obtained product W2 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0123] 1 H NMR (400MHz, CD3OD) δ8.29(d,J=6.9Hz,1H),7.90(d,J=7.8Hz,2H),7.84(t,J=7.9 Hz,2H),7.49(q,J=6.8Hz,2H),7.43–7.26(m,8H),6.26(d,J=9.7Hz,1H),6.03(t,J =9.9Hz,1H),5.78(d,J=8.5Hz,1H),5.21(d,J=3.8Hz,1H),4.62(s,2H),4.42(dd, J=33.8,8.0Hz,2H),4.10–3.94(m,2H),3.83–3.69(m,1H),2.97(d,J=19.2Hz,3H). 13 C NMR(101MHz,CD3OD)δ170.10,165.27,157.40,149.35,141.85,139.50,13 6.53,133.59,129.49,129.43,128.23,128.19,128.11,127.72,127.51,12 7.34,125.13,91.83,72.26,70.76,67.45,61.16,55.88,51.81,50.87,48. 27,48.05,47.85,47.84,47.63,47.54,47.41,47.20,46.99,35.11,29.35. 19 F NMR(376MHz,CD3OD)δ-166.64.ESI-HRMS(m / z):calcd for C 38 H 37 F6O 13 :[M+Na] + 827.22948;found:827.22973.[a] D 20 +32.6 (c 0.12, DMF).

[0124] 5. Synthesize compound 7-1 or W3, the chemical reaction equation is:

[0125]

[0126] The specific synthesis process is:

[0127] Compound 6-1 (543 mg, 0.66 mmol) was dissolved in CH3OH (12 mL), and an aqueous solution of LiOH (32 mg, 1.32 mmol) (12 mL) was added dropwise at 0°C. After stirring at 0°C for 3 hours, the reaction of the raw material was complete as monitored by TLC. AcOH was added dropwise to adjust the solution pH to 2. The reaction solution was concentrated and filtered with a micromembrane filter. The filtrate was filtered with C 18 Column, separated by preparative HPLC, chromatographic conditions: 13% CH3CN + 1% AcOH aqueous solution; flow rate: 5mL / min; detection by UV detector, compound 7-1 retention time t = 120min, compound W3 retention time t = 150min, to obtain two compounds 7-1 (108mg, 27%) and W3 (191mg, 48%), both as white solids.

[0128] The obtained product 7-1 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0129] 1 H NMR(400MHz,CH3OD)δ8.09(d,J=6.5Hz,1H),7.44–7.29(m,5H),5.60(s,1H),5.16(d,J=19.1 Hz, 2H), 4.50 (s, 1H), 4.07 (d, J = 11.5Hz, 4H), 3.73 (d, J = 14.0Hz, 4H), 3.03 (d, J = 22.0Hz, 3H). 13 C NMR(151MHz,D2O)δ176.62,174.61,171.76,171.64,159.18,159.01,158.22,157.74,150.36,141.74,135.99,1 28.68,128.28,127.60,127.39,125.63,82.29,75.54,73.06,71.02,67.77,61.60,61.20,55.45,52.72,35.76. 19 F NMR(376MHz,CD3OD)δ-168.18.ESI-HRMS(m / z):calcd for C 24 H 28 F5O 12 :[M+Na] +620.16107; found:620.16102.[a] D 20 +15.6 (c 0.14, DMF).

[0130] The obtained product W3 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0131] 1 H NMR(400MHz,CH3OD)δ8.09(d,J=6.5Hz,1H),7.44–7.29(m,5H),5.60(s,1H),5.20–5.10 (m,2H),4.50(s,1H),4.07(d,J=11.5Hz,4H),3.84–3.66(m,4H),3.03(d,J=22.0Hz,3H). 13 CNMR(151MHz,D2O)δ176.66,174.61,171.76,171.64,159.18,159.01,15 8.22,157.74,150.36,141.74,140.19,135.99,128.68,128.28,127.60, 127.39,82.29,75.54,73.06,71.02,67.77,61.60,61.20,61.08,55.45, 52.72,51.88,51.39,35.76,20.38,20.26,13.11.ESI-HRMS(m / z):calcd for C 24 H 28 F5O 12 :[M+Na] + 620.16107;found:620.16085.[a] D 20 +13.5 (c 0.13, DMF).

[0132] 6. Synthesize compound 9-1, the chemical reaction equation is:

[0133]

[0134] The specific synthesis process is:

[0135] Compound 7-1 (50 mg, 0.0837 mmol) was dissolved in EtOH (2 mL) and water (2 mL), and Pd / C (20 mg) was added. After reacting at room temperature for 30 minutes under hydrogen, the reaction was complete after TLC monitoring. The solvent ethanol was dried and dissolved in a small amount of water, and filtered with a micromembrane filter. The filtrate was filtered with C 18Column, separated by preparative HPLC, chromatographic conditions: 5% CH3CN + 1% AcOH aqueous solution; flow rate: 5mL / min; UV detector detection, compound 9-1 retention time t = 15min, the eluate was concentrated under reduced pressure to obtain pyrimidine nucleoside peptide compound 9-1 (33mg, 86%).

[0136] The obtained product 9-1 was measured on a nuclear magnetic resonance instrument, and the nuclear magnetic resonance data obtained were:

[0137] 1 H NMR (400MHz, D2O) δ8.01(d,J=6.2Hz,1H),5.54(d,J=8.5Hz,1H),4.42(t,J=5.3Hz,1H),3.97–3.84(m,4H),3.76–3.62(m,4H),2.67(s,3H). 13 C NMR(101MHz,D2O)δ178.69,173.91,171.22,166.32,159.30,159.05,150.46,142.16,139. 84,126.01,125.66,82.52,77.68,73.49,71.43,61.43,55.74,53.21,49.37,32.83,21.59. 19 F NMR(376MHz,D2O)δ-165.11.ESI-HRMS(m / z):calcd for C 16 H 22 F5O 10 :[M+Na] + 486.12429; found:486.12409.[a] D 20 +35.4 (c 0.12, DMSO).

[0138] 7. Synthesize compound W4, the chemical reaction equation is:

[0139]

[0140] The specific synthesis process is:

[0141] Compound W3 (50 mg, 0.0837 mmol) was dissolved in EtOH (2 mL) and water (2 mL), and Pd / C (20 mg) was added. After reacting at room temperature for 30 minutes under hydrogen, the reaction was complete after TLC monitoring. The solvent ethanol was dried and dissolved in a small amount of water, and filtered with a micromembrane filter. The filtrate was filtered with C 18Column, separated by preparative HPLC, chromatographic conditions: 5% CH3CN + 1% AcOH aqueous solution; flow rate: 5mL / min; detection by UV detector, compound W4 retention time t = 15min, the eluate was concentrated under reduced pressure to obtain pyrimidine nucleoside peptide compound W4 (34mg, 90%).

[0142] The obtained product W4 was measured on a nuclear magnetic resonance instrument, and the nuclear magnetic resonance data obtained were:

[0143] 1 H NMR(400MHz,D2O)δ8.03(d,J=6.2Hz,1H),5.55(d,J=8.8Hz,1H),4.45(t,J=5.2Hz,1 H),3.97–3.87(m,4H),3.79–3.75(m,2H),3.69(dd,J=13.3,9.0Hz,2H),2.70(s,3H). 13 C NMR(101MHz,D2O)δ178.33,173.83,171.32,166.42,159.35,159.09,150.49,142.20,139. 87,125.99,125.64,82.51,77.65,73.40,71.58,61.28,55.64,53.29,49.47,32.83,21.36. 19 F NMR(376MHz,D2O)δ-165.13.ESI-HRMS(m / z):calcd for C 16 H 22 F5O 10 :[M+H] + 464.14235; found:464.14348.[a] D 20 +24.8 (c 0.11, DMSO).

[0144] Example 2 Total Synthesis of W5-W6

[0145] 1. Synthesize compound 3-2, the chemical reaction equation is:

[0146]

[0147] The specific synthesis process is:

[0148] Add 5-I substituted uracil (BS2) (1.91 g, 8.04 mmol) and BSA (3.55 mL, 21.44 mmol) to dry CH3CN (50 mL), stir at room temperature for 10 minutes, and the reaction solution changes from turbid to clear. Add compound 1 (3.0 g, 5.36 mmol), followed by TMSOTf (3.50 mL, 29.59 mmol). Under N2 protection, react at 77 ° C, and TLC monitors the reaction to be complete. Concentrate, extract the reaction solution with water and ethyl acetate (3×200 mL). Combine the ethyl acetate layers, wash with saturated NaCl solution (2×200 mL), dry over anhydrous Na2SO4, filter, and concentrate under reduced pressure to obtain a crude nucleoside compound.

[0149] The crude compound (3.95 g, 5.36 mmol) was dissolved in anhydrous CH3OH (40 mL), and NaOH (0.21 g, 5.36 mmol) was added. After reacting at room temperature for 1-3 hours, the reaction was complete as monitored by TLC, and the 6-position protecting group was selectively removed. 1M HCl was added to quench the reaction, and the concentrate was concentrated under reduced pressure. The obtained concentrate was separated by silica gel column chromatography (dichloromethane / methanol, 40:1) to obtain a white solid compound 3-2 (1.17 g, 35%, two steps).

[0150] The obtained compound 3-2 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0151] 1 H NMR(400MHz, CDCl3)δ9.51(s,1H),8.03–7.91(m,3H),7.87–7.81(m,2H),7.57–7.43(m,2H),7.38(t,J=7.8Hz,2H),7.34–7.27(m,2H),6.12(d ,J=9.4Hz,1H),5.90(t,J=9.7Hz,1H),5.58(t,J=9.2Hz,1H),4.19–4.02(m,3H),3.95(dd,J=12.5,4.1Hz,1H),3.86–3.77(m,1H),2.07(s,1H). 13 C NMR (151MHz, CDCl3) δ165.50,165.46,159.96,150.10,144.50,133.96,133.75,130.03,129. 85,128.57,128.46,127.76,77.84,73.73,70.45,70.01,61.34,59.47.ESI-HRMS(m / z):calcd for C 24 H 20 IN5O8:[M+Na]+ 656.02488; found 656.02566.

[0152] 2. Synthesize compound 4-2, the chemical reaction equation is:

[0153]

[0154] The specific synthesis process is:

[0155] Compound 3-2 (1.17 g, 1.85 mmol) was dissolved in CH3CN (6 mL) and water (4 mL), TEMPO (88 mg, 0.56 mmol) was added, and iodobenzene acetate (1.48 g, 5.55 mmol) was added in batches. After stirring at room temperature for 6 hours, the reaction was complete after TLC monitoring. CH3OH (3 mL) was added to quench the reaction, and the mixture was concentrated and briefly slurried with n-hexane to obtain the carboxylic acid product, which could be directly used for the next step.

[0156] The above crude product was dissolved in anhydrous CH3OH (14mL), H2SO4 (0.37mL) was added, and the reaction was carried out at room temperature. The reaction was completed after TLC monitoring. NaHCO3 was added to quench the reaction and concentrated. The reaction solution was extracted with water (40mL) and ethyl acetate (3×100mL), and the combined ethyl acetate solution was washed with saturated NaCl solution (2×50mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain a crude product of the 6-position methyl ester of the sugar ring, which can be directly used for the next step reaction.

[0157] SnCl2·2H2O (0.83 g, 3.70 mmol) and p-methylthiophenol (0.92 g, 7.40 mmol) were added to CHCl3 (10 mL), and Et3N (1.54 mL, 11.10 mmol) was added dropwise at room temperature. Stir for 5 minutes, and the above crude methyl ester (1.22 g, 1.85 mmol) was added. The reaction was carried out at room temperature, and the reaction was complete when monitored by TLC. The solvent was dried, ethyl acetate was added to dissolve, and the insoluble matter SnCl2·2H2O was removed by filtration. The mixture was dried, and silica gel was added to mix the sample. The mixture was separated by silica gel column chromatography (CH2Cl2 / CH3OH, 40:1) to obtain a white solid compound 4-2 (0.88 g, 75%, three steps).

[0158] The intermediate 4-2 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0159] 1H NMR (400MHz, DMSO-d6) δ8.52(s,1H),7.90(d,J=7.6Hz,2H),7.74(d,J=7.6Hz,2H),7.60(dd,J=7.5,3.2Hz,2H),7.46(dt,J= 12.1,7.7Hz,4H),6.14(d,J=9.2Hz,1H),5.81(t,J=9.2Hz,1H),5.67(t,J=9.5Hz,1H),4.49(d,J=10.1Hz,1H),3.74(s,3H). 13 CNMR(151MHz,DMSO-d6)δ168.69,165.96,165.10,160.56,150.53,145.73,134.37,133.96,129.75,129 .72,129.57,129.29,129.10,128.69,78.38,76.02,71.52,71.38,53.23,52.77.ESI-HRMS(m / z):calcd for C 25 H 22 IN3O9:[M+H] + 636.04735; found:636.04832.

[0160] 3. Synthesize compound 6-2, the chemical reaction equation is:

[0161]

[0162] The specific synthesis process is:

[0163] The above compound 5-1 (860 mg, 2.77 mmol) was dissolved in dry DMF (20 mL), and then HATU (1.92 g, 5.04 mmol) and 2,6-dimethylpyridine (1.22 mL, 10.46 mmol) were added in sequence, and stirred at room temperature for 2 hours. Compound 4-2 (800 mg, 1.26 mmol) was added, and stirring was continued at room temperature for 14 hours. After TLC monitoring, the reaction of the raw materials was complete. Saturated NaCl solution (30 mL) was added, and the reaction solution was extracted with ethyl acetate (3×100 mL). The combined ethyl acetate solution was washed with saturated NaCl solution (2×50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The obtained concentrate was separated by silica gel column chromatography (CH2Cl2 / CH3OH, 50:1) to obtain a light yellow solid compound 6-2 (473 mg, 40%).

[0164] The obtained product 6-2 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0165] 1 H NMR (400MHz, DMSO-d6) δ11.76(s,1H),8.74(s,1H),8.15(s,1H),8.00(dd,J=12.2,7.9Hz,1H),7.7 7(qd,J=5.5,4.8,2.4Hz,4H),7.66–7.54(m,2H),7.51–7.40(m,4H),7.39–7.24(m,8H),6.16(d,J=9 .8Hz,1H),5.95(t,J=9.2Hz,1H),5.82–5.71(m,1H),5.13–4.96(m,3H),4.79(q,J=5.7,5.2Hz,1H), 4.62(d,J=10.2Hz,1H),4.34–4.16(m,1H),3.95–3.88(m,1H),3.64(s,3H),2.66(d,J=14.8Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ170.69,168.41,167.54,165.75,165.58,164.99,160.55,156 .37,156.07,150.54,137.42,134.37,133.89,129.65,129.30,129.04,128.86,128.2 5,128.04,127.85,127.55,75.17,73.27,73.00,71.51,71.24,66.69,61.91,55.93,5 2.84,49.87,40.35,40.21,40.07,39.93,39.79,39.65,39.51.ESI-HRMS(m / z):calcd for C 39 H 38 IN5O 14 :[M+H] + 928.15327; found:928.15637.[a] D 20 -18.5 (c 0.10, DMF).

[0166] 4. Synthesize compound W5 or 8-2, the chemical reaction equation is:

[0167]

[0168] The specific synthesis process is:

[0169] Compound 6-2 (456 mg, 0.49 mmol) was dissolved in CH3OH (15 mL), and an aqueous solution of LiOH (9 mg, 0.39 mmol) (9 mL) was added dropwise at 0°C. After stirring at 0°C for 3 hours, the reaction of the raw material was complete as monitored by TLC. AcOH was added dropwise to adjust the solution pH to 2. The reaction solution was concentrated and filtered with a micromembrane filter. The filtrate was filtered with C 18 Column, separated by preparative HPLC, chromatographic conditions: 14% CH3CN + 1% AcOH aqueous solution; flow rate: 5 mL / min; detection by UV detector, compound W5 retention time t = 250 min, compound 8-2 retention time t = 300 min, to obtain two compounds W5 (62 mg, 18%) and 8-2 (52 mg, 15%), both as white solids.

[0170] The obtained product W5 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0171] 1 H NMR(400MHz,D2O)δ8.16(s,1H),7.32–7.15(m,5H),5.45(d,J=7.4Hz,1H),4.99(dd,J=26.8,7. 9Hz, 2H), 4.33 (d, J = 24.6Hz, 1H), 3.91 (s, 2H), 3.62 (t, J = 22.0Hz, 4H), 2.84 (d, J = 24.5Hz, 3H). 13 C NMR(151MHz,DMSO-d6)δ170.92,170.17,168.72,160.81,150.99,146.00,128.91,128.83,128.30,127.90,1 27.63,126.59,79.58,76.77,70.80,66.58,60.27,55.74,52.61,40.76,21.22,14.54.ESI-HRMS(m / z):calcd for C 24 H 28 IN5O 12 :[M+Na] + 728.06714; found:728.06900.[a] D 20 +5.9 (c 0.12, DMF).

[0172] 5. Synthesize compound W6, the chemical reaction equation is:

[0173]

[0174] The above compound 5-2 (synthesis method see reference: Org.Biomol.Chem.2021,19,4285-4291, or patent: CN107973830B) (370mg, 1.32mmol) was dissolved in dry DMF (210mL), followed by the addition of HATU (0.91g, 2.4mmol) and 2,6-dimethylpyridine (0.58mL, 4.98mmol), and stirred at room temperature for 2 hours. Compound 4-2 (381mg, 0.6mmol) was added, and stirring continued at room temperature for 14 hours. TLC monitored the complete reaction of the raw materials. Saturated NaCl solution (30mL) was added, and the reaction solution was extracted with ethyl acetate (3×50mL). The combined ethyl acetate solution was washed with saturated NaCl solution (2×50mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The obtained concentrate was separated by silica gel column chromatography (CH2Cl2 / CH3OH, 50:1) to obtain a light yellow solid compound;

[0175] The above light yellow solid compound (243 mg, 0.27 mmol) was dissolved in MeOH (2 mL), and an aqueous solution (1 mL) of LiOH (8 mg, 0.32 mmol) was added dropwise at 0°C. After stirring at 0°C for 3 hours, the reaction of the raw material was complete as monitored by TLC. AcOH was added dropwise to adjust the solution pH to 2. The reaction solution was concentrated and filtered with a micromembrane filter. The filtrate was filtered with C 18 Column, separated by preparative HPLC, chromatographic conditions: 20% CH3CN + 1% AcOH aqueous solution; flow rate: 5mL / min; UV detector detection, retention time t = 90min, to obtain a white solid compound W6 (48mg, 26%).

[0176] The obtained product W6 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0177] 1 H NMR(600MHz,DMSO-d6)δ11.76(s,1H),8.25(s,1H),8.09(s,1H),7.39–7.22(m,5H),5.73(s,1H),5 .59–5.49(m,1H),5.38–5.24(m,2H),3.96–3.83(m,4H),3.71–3.60(m,2H),2.85(d,J=25.5Hz,3H). 13C NMR(151MHz,DMSO-d6)δ172.53,169.38,169.12,168.83,160.80,156.47,156.13,150.99,137.46,137.36,12 8.88,128.79,127.90,76.44,73.90,71.07,70.92,66.75,66.55,52.71,42.14,21.53.ESI-HRMS(m / z):calcd for C 23 H 26 IN5O 11 :[M+Na] + 698.05657; found:698.05714.[a] D 20 10.3 (c0.11, DMF).

[0178] Example 3 Total Synthesis of W7-W8

[0179] 1. Synthesize compound 3-3, the chemical reaction equation is:

[0180]

[0181] The specific synthesis process is:

[0182] 5-CH3 substituted uracil (BS3) (1.69 g, 13.41 mmol) and BSA (5.92 mL, 35.76 mmol) were added to dry CH3CN (50 mL), stirred at room temperature for 30 minutes, and the reaction solution changed from turbid to clear. Compound 1 (5.0 g, 8.94 mmol) was added, followed by TMSOTf (5.84 mL, 32.18 mmol). Under N2 protection, the reaction was carried out at 77 °C, and the reaction was complete when monitored by TLC. Concentrate, extract the reaction solution with water and ethyl acetate (3 × 200 mL). Combine the ethyl acetate layers, wash with saturated NaCl solution (2 × 200 mL), dry over anhydrous Na2SO4, filter, and concentrate under reduced pressure to obtain a crude nucleoside compound.

[0183] The crude nucleoside compound (5.59 g, 8.94 mmol) was dissolved in anhydrous CH3OH (50 mL), and NaOH (0.36 g, 8.94 mmol) was added. After reacting at room temperature for 1-3 hours, the reaction was complete as monitored by TLC, and the 6-position protecting group was selectively removed. 1M HCl was added to quench the reaction, and the concentrate was concentrated under reduced pressure. The obtained concentrate was separated by silica gel column chromatography (dichloromethane / methanol, 40:1) to obtain a white solid compound 3-3 (2.61 g, 56%, two steps).

[0184] The obtained compound 3-3 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0185] 1 H NMR(400MHz,CH3OD)δ8.02–7.96(m,2H),7.89–7.79(m,3H),7.61–7.51(m,2H),7.41(dt,J=25.1,7.8Hz,4H),6.17(d,J=9 .2Hz,1H),5.94(t,J=9.7Hz,1H),5.69(t,J=9.4Hz,1H),4.23(t,J=10.0Hz,1H),3.99–3.80(m,3H),1.92(d,J=1.2Hz,3H). 13 C NMR(101MHz,CD3OD)δ165.52,165.21,164.53,150.84,136.62,133.49,133.46,129.40,129.33,128.77, 128.30,128.26,128.22,110.88,80.38,77.53,74.20,70.85,60.42,59.25,10.97.ESI-HRMS(m / z):calcd for C 25 H 23 N5O8:[M+Na] + 544.14388; found:544.14374.

[0186] 2. Synthesize compound 4-3, the chemical reaction equation is:

[0187]

[0188] The specific synthesis process is:

[0189] Compound 3-3 (2.61 g, 5.0 mmol) was dissolved in CH3CN (13 mL) and water (9 mL), TEMPO (234 mg, 1.5 mmol) was added, and iodobenzene acetate (4.0 g, 15 mmol) was added in batches. After stirring at room temperature for 6 hours, the reaction was complete after TLC monitoring. CH3OH (5 mL) was added to quench the reaction, and the mixture was concentrated and briefly slurried with n-hexane to obtain the carboxylic acid product, which could be directly used for the next step.

[0190] The crude carboxylic acid product was dissolved in anhydrous CH3OH (40 mL), H2SO4 (1.0 mL) was added, and the reaction was carried out at room temperature. The reaction was completed after TLC monitoring. NaHCO3 was added to quench the reaction and concentrated. The reaction solution was extracted with water (40 mL) and ethyl acetate (3×100 mL), and the combined ethyl acetate solution was washed with saturated NaCl solution (2×50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain a crude product of the 6-position methyl ester of the sugar ring, which can be directly used for the next step reaction.

[0191] SnCl2·2H2O (2.26 g, 10.0 mmol) and p-methylthiophenol (2.48 g, 20.0 mmol) were added to CHCl3 (35 mL), and Et3N (4.17 mL, 30.0 mmol) was added dropwise at room temperature. Stir for 5 minutes, and the above crude methyl ester (2.75 g, 5.0 mmol) was added. The reaction was carried out at room temperature, and the reaction was complete when monitored by TLC. The solvent was dried by spin drying, ethyl acetate was added to dissolve, and the insoluble matter SnCl2·2H2O was filtered out, dried by spin drying, and silica gel was added to mix the sample. The white solid compound 4-3 (2.07 g, 79%) was obtained by silica gel column chromatography (CH2Cl2 / CH3OH, 40:1).

[0192] The obtained intermediate 4-3 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0193] 1 H NMR (400MHz, DMSO-d6) δ11.36(s,1H),7.94–7.84(m,3H),7.77–7.71(m,2H),7.61(td,J=7.4,3.6Hz,2H),7.46( dt,J=12.8,7.7Hz,4H),6.21–6.12(m,1H),5.74–5.63(m,2H),4.49(d,J=10.1Hz,1H),3.74(s,3H),1.81(s,3H). 13 C NMR (101MHz, DMSO) δ168.76,165.96,164.98,163.83,150.92,134.33,133.93,129.78,129.74,129.5 5,129.29,129.10,128.78,110.71,78.54,76.30,71.32,53.73,52.74,12.52.ESI-HRMS(m / z):calcd forC 26 H 25 N3O9:[M+Na] + 546.14830; found:546.14809.

[0194] 3. Synthesize compound 6-3, the chemical reaction equation is:

[0195]

[0196] The specific synthesis process is:

[0197] The above compound 5-1 (1.31 g, 4.2 mmol) was dissolved in dry DMF (20 mL), and then HATU (2.91 g, 7.64 mmol) and 2,6-dimethylpyridine (1.85 mL, 15.85 mmol) were added in sequence, and stirred at room temperature for 2 hours. Compound 4-3 (1.0 g, 1.91 mmol) was added, and stirring was continued at room temperature for 14 hours. After TLC monitoring, the reaction of the raw materials was complete. Saturated NaCl solution (50 mL) was added, and the reaction solution was extracted with ethyl acetate (3×100 mL). The combined ethyl acetate solution was washed with saturated NaCl solution (2×50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The obtained concentrate was separated by silica gel column chromatography (CH2Cl2 / CH3OH, 50:1) to obtain a light yellow solid compound 6-3 (402 mg, 26%).

[0198] The obtained product 6-3 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0199] 1 H NMR (400MHz, CDCl3) δ7.98–7.75(m,5H),7.50(d,J=7.6Hz,1H),7.36(q,J=14.1, 8.6Hz,9H),7.16(dd,J=14.7,7.6Hz,2H),6.35(t,J=8.9Hz,1H),5.81(m,J=38.1 ,9.5Hz,1H),5.18(d,J=26.6Hz,2H),4.67(d,J=10.1Hz,1H),4.51–4.32(m,1H), 4.01(s,2H),3.80(d,J=12.0Hz,4H),2.98(d,J=30.2Hz,3H),2.08–1.65(m,6H). 13C NMR (151MHz, CDCl3) δ171.45,171.14,170.18,169.64,168.15,167.73,166. 49,166.34,165.17,163.24,157.29,150.77,150.62,136.23,134.71,133.7 1,129.82,128.55,128.40,127.82,112.51,80.00,75.47,69.71,67.83,62. 47,61.56,54.28,53.35,36.10,29.70,12.41,12.20.ESI-HRMS(m / z):calcd for C 40 H 41 N5O 14 :[M+Na] + 838.25422; found:838.25606.[a] D 20 +7.1(c0.11,DMF).

[0200] 4. Synthesize compound W7 or W8, the chemical reaction equation is:

[0201]

[0202] The specific synthesis process is:

[0203] Compound 6-3 (249 mg, 0.31 mmol) was dissolved in CH3OH (15 mL), and an aqueous solution (10 mL) of LiOH (19 mg, 0.78 mmol) was added dropwise at 0°C. After stirring at 0°C for 3 hours, the reaction of the raw material was complete as monitored by TLC. AcOH was added dropwise to adjust the solution pH to 2. The reaction solution was concentrated and filtered with a micromembrane filter. The filtrate was filtered with C 18 Column, separated by preparative HPLC, chromatographic conditions: 12% CH3CN + 1% AcOH aqueous solution; flow rate: 5 mL / min; detection by UV detector, compound W7 retention time t = 230 min, compound W8 retention time t = 280 min, to obtain two compounds W7 (8.3 mg, 5%) and W8 (13.2 mg, 7%), both as white solids.

[0204] The obtained product W7 was identified by nuclear magnetic resonance hydrogen spectrum, high-resolution mass spectrometer and polarimeter, and the obtained data were:

[0205] 1H NMR(400MHz,D2O)δ7.69(s,1H),7.46–7.31(m,5H),5.64(d,J=7.4Hz,1H),5.19–5.06(m,2H),4.57–4.41(m,1H ),4.09(d,J=18.5Hz,4H),3.87–3.68(m,4H),2.99(dd,J=22.7,3.3Hz,3H),1.89(s,3H).ESI-HRMS(m / z):calcd for C 25 H 31 N5O 12 :[M+Na] + 616.18614; found:616.18658.[a] D 20 +13.7 (c 0.10, DMF).

[0206] The obtained product W8 was identified by a high-resolution mass spectrometer and a polarimeter, and the obtained high-resolution mass spectrum and specific rotation data were:

[0207] ESI-HRMS(m / z):calcd for C 25 H 31 N5O 12 :[M+Na] + 616.18614; found:616.18566.[a] D 20 +16.3(c0.10,DMF).

[0208] 5. Synthesize compound 9-3, the chemical reaction equation is:

[0209]

[0210] The specific synthesis process is:

[0211] Compound W7 (39 mg, 0.0657 mmol) was dissolved in EtOH (4 mL) and water (4 mL), and Pd / C (16 mg) was added. The mixture was reacted at room temperature under hydrogen for 30 minutes. The reaction was complete after TLC monitoring. The filtrate was filtered and the filtrate was washed with C 18 Column, separated by preparative HPLC, chromatographic conditions: 6% CH3CN + 1% AcOH aqueous solution; flow rate: 5mL / min; UV detector detection, compound 9-3 retention time t = 15min, the eluate was concentrated under reduced pressure to obtain pyrimidine nucleoside peptide compound 9-3 (26mg, 86%).

[0212] The obtained product 9-3 was measured on a nuclear magnetic resonance instrument, and the nuclear magnetic resonance data obtained were:

[0213] 1 H NMR(400MHz,D2O)δ7.57(s,1H),5.46(dd,J=8.6,3.5Hz,1H),4.22(q,J=5.1 Hz,1H),3.85(d,J=6.5Hz,2H),3.76–3.59(m,6H),2.79(s,3H),1.74(s,3H). 13 C NMR (101MHz, D2O) δ173.15,166.17,151.98,137.54,112.00,82.21,77.66,73.67,71.21,62.67,61.97,56.63,53.37,35.16,11.42.[a] D 20 +4.3 (c 0.14, DMSO).

[0214] 6. Synthesize compound 10-3, the chemical reaction equation is:

[0215]

[0216] The specific synthesis process is:

[0217] Compound W8 (39 mg, 0.0657 mmol) was dissolved in EtOH (4 mL) and water (4 mL), and Pd / C (16 mg) was added. The mixture was reacted at room temperature under hydrogen for 30 minutes. The reaction was complete after TLC monitoring. The filtrate was filtered and the filtrate was washed with C 18 Column, separated by preparative HPLC, chromatographic conditions: 6% CH3CN + 1% AcOH aqueous solution; flow rate: 5mL / min; UV detector detection, compound 10-3 retention time t = 15min, the eluate was concentrated under pressure to obtain pyrimidine nucleoside peptide compound 10-3 (15.2mg, 50%).

[0218] The obtained product 10-3 was measured on a nuclear magnetic resonance instrument, and the nuclear magnetic resonance data obtained were:

[0219] 1 H NMR(400MHz,D2O)δ7.65(s,1H),5.56–5.49(m,1H),4.45(t,J=5.3Hz,1H),3.97 –3.83(m,5H),3.78–3.73(m,2H),3.73–3.68(m,2H),2.69(s,3H),1.82(s,3H). 13C NMR(101MHz,D2O)δ179.48,171.95,166.41,166.24,152.04,137.46,112.09,82.22,7 9.19,73.52,71.51,61.27,55.63,53.38,49.46,32.80,11.44.ESI-HRMS(m / z):calcd forC 17 H 25 N5O 10 :[M+H] + 460.16742;found:460.16845.[a] D 20 +8.1 (c 0.10, DMSO).

[0220] Example 4 Total Synthesis of W9-W12

[0221] 1. Synthesize compound 3-4, the chemical reaction equation is:

[0222]

[0223] The specific synthesis process is:

[0224] 5-CF3 substituted uracil (BS4) (2.22 g, 12.33 mmol) and BSA (5.45 mL, 32.88 mmol) were added to dry CH3CN (80 mL), stirred at room temperature for 10 minutes, and the reaction solution changed from turbid to clear. Compound 1 (4.60 g, 8.22 mmol) was added, followed by TMSOTf (5.37 mL, 29.59 mmol). Under N2 protection, the reaction was carried out at 77 °C, and the reaction was monitored by TLC to be complete. Concentrate, extract the reaction solution with water and ethyl acetate (3 × 200 mL). Combine the ethyl acetate layers, wash with saturated NaCl solution (2 × 200 mL), dry over anhydrous Na2SO4, filter, and concentrate under reduced pressure to obtain a crude nucleoside compound.

[0225] The crude nucleoside compound (5.59 g, 8.22 mmol) was dissolved in anhydrous CH3OH (30 mL), and NaOH (0.33 g, 8.22 mmol) was added. After reacting at room temperature for 1-3 hours, the reaction was complete as monitored by TLC, and the 6-position protecting group was selectively removed. 1M HCl was added to quench the reaction, and the concentrate was concentrated under reduced pressure. The obtained concentrate was separated by silica gel column chromatography (dichloromethane / methanol, 40:1) to obtain a white solid compound 3-4 (1.84 g, 39%, two steps).

[0226] The obtained compound 3-4 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0227] 1 H NMR (400MHz, DMSO-d6) δ11.99(br,1H),8.61(s,1H),7.94(dd,J=8.1,1.5Hz,2H),7.81–7. 74(m,2H),7.63(dt,J=10.4,7.5Hz,2H),7.48(dt,J=17.9,7.7Hz,4H),6.35(d,J=9.0Hz,1H ),6.04(t,J=9.6Hz,1H),5.79(t,J=9.2Hz,1H),5.14–5.07(m,1H),4.24(t,J=10.0Hz,1H), 4.07(ddd,J=10.2,4.8,1.9Hz,1H), 3.77(ddd,J=12.4,5.1,1.8Hz,1H), 3.70–3.61(m,1H). 13 C NMR(101MHz,DMSO-d6)δ165.50,165.39,158.86,149.84,134.49,134.44,129.73,129.69,129 .39,129.30,129.03,128.98,128.50,77.05,73.82,71.94,60.82,60.23,59.36,21.22,14.53. 19 F NMR(376MHz,DMSO-d6)δ-61.58.ESI-HRMS(m / z):calcd for C 26 H 20 F3N5O9:[M+Na] + 598.11562; found 598.11547.

[0228] 2. Synthesize compound 4-4, the chemical reaction equation is:

[0229]

[0230] The specific synthesis process is:

[0231] Compound 3-4 (1.36 g, 2.37 mmol) was dissolved in CH3CN (15 mL) and water (12 mL), TEMPO (111 mg, 0.71 mmol) was added, and iodobenzene acetate (1.89 g, 7.11 mmol) was added in batches. After stirring at room temperature for 6 hours, the reaction was complete after TLC monitoring. CH3OH (5 mL) was added to quench the reaction, and the mixture was concentrated and slurried with n-hexane to obtain the carboxylic acid product, which could be directly used for the next step.

[0232] The above crude product was dissolved in anhydrous CH3OH (20mL), H2SO4 (0.48mL) was added, and the reaction was carried out at room temperature. The reaction was completed by TLC monitoring. NaHCO3 was added to quench the reaction and concentrated. The reaction solution was extracted with water (40mL) and ethyl acetate (3×50mL), and the combined ethyl acetate solution was washed with saturated NaCl solution (2×50mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain a crude product of the 6-position methyl ester of the sugar ring, which can be directly used for the next step reaction.

[0233] SnCl2·2H2O (1.07 g, 4.74 mmol) and p-methylthiophenol (1.18 g, 9.48 mmol) were added to CHCl3 (25 mL), and Et3N (1.32 mL, 9.48 mmol) was added dropwise at room temperature. Stir for 5 minutes, and the above crude methyl ester (1.43 g, 2.37 mmol) was added. The reaction was carried out at room temperature, and the reaction was monitored by TLC. The solvent was dried, ethyl acetate was added to dissolve, the insoluble matter SnCl2·2H2O was filtered out, dried, silica gel was added to mix the sample, and separated by silica gel column chromatography (CH2Cl2 / CH3OH, 40:1) to obtain white solid compound 4-4 (1.11 g, 81%, three steps).

[0234] The obtained intermediate 4-4 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0235] 1 H NMR (400MHz, DMSO-d6) δ8.67(s,1H),7.90(d,J=7.5Hz,2H),7.77–7.72(m,2H),7.60(d,J=7.5Hz,2H),7.47(t,J= 8.0Hz, 4H), 6.22 (d, J = 9.2Hz, 1H), 5.89 (t, J = 9.2Hz, 1H), 5.78–5.67 (m, 1H), 4.55 (d, J = 10.1Hz, 1H), 3.74 (s, 3H). 13 C NMR(151MHz,DMSO-d6)δ168.55,165.95,165.27,158.82,149.85,143.52,134.39,133.9 6,129.73,129.59,129.29,129.10,128.63,121.90,78.32,75.70,71.61,53.05,52.80. 19 F NMR(376MHz,DMSO-d6)δ-61.54.ESI-HRMS(m / z):calcd for C 26 H 22 F3N3O9:[M+H]+ 578.13809; found:578.13825.

[0236] 3. Synthesize compound 6-4, the chemical reaction equation is:

[0237]

[0238] The specific synthesis process is:

[0239] The above compound 5-1 (200 mg, 0.64 mmol) was dissolved in dry DMF (5 mL), and then HATU (395.45 mg, 1.04 mmol) and 2,6-dimethylpyridine (0.25 mL, 2.16 mmol) were added in sequence, and stirred at room temperature for 2 hours. Compound 4-4 (150.14 mg, 0.26 mmol) was added, and stirring was continued at room temperature for 14 hours. After TLC monitoring, the reaction of the raw materials was complete. Saturated NaCl solution (30 mL) was added, and the reaction solution was extracted with ethyl acetate (3×50 mL). The combined ethyl acetate solution was washed with saturated NaCl solution (2×30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The obtained concentrate was separated by silica gel column chromatography (CH2Cl2 / CH3OH, 50:1) to obtain a light yellow solid compound 6-4 (120 mg, 53%).

[0240] The obtained product 6-4 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0241] 1 H NMR (400MHz, CDCl3) δ10.22(s,1H),8.18(d,J=27.9Hz,1H),7.94–7.74(m,4H),7.48(t,J=7.4Hz,1H),7.40–7.30(m,8H),7.14(t,J=7.8Hz,2H),6.3 8(d,J=9.5Hz,1H),6.08(t,J=9.8Hz,1H),5.16(d,J=11.7Hz,3H),4.75(d ,J=7.7Hz,1H),4.06–3.90(m,2H),3.82(s,3H),2.92(s,2H),2.09(s,3H). 13C NMR (151MHz, CDCl3) δ171.24,170.43,167.44,166.21,165.35,157.82,157.34,14 9.74,136.16,133.83,129.84,129.76,128.57,128.53,128.48,128.20,128.14,1 27.83,127.74,127.69,122.36,120.56,107.08,106.86,80.37,77.25,77.04,76.82,75.41,72.57,69.91,67.91,61.52,54.26,53.40,53.04,51.54,36.04,29.71. 19 F NMR(376MHz,CDCl3)δ-63.57.ESI-HRMS(m / z):calcd for C 40 H 38 F3N5O 14 :[M+Na] + 892.22596;found:892.22803.[a] D 20 +8.4 (c 0.10, DMF).

[0242] 4. Synthesize compound W9 or W10, the chemical reaction equation is:

[0243]

[0244] The specific synthesis process is:

[0245] Compound 6-4 (200 mg, 0.23 mmol) was dissolved in CH3OH (5 mL), and an aqueous solution (3 mL) of LiOH (17 mg, 0.69 mmol) was added dropwise at 0°C. After stirring at 0°C for 3 hours, the reaction of the raw material was complete as monitored by TLC. AcOH was added dropwise to adjust the solution pH to 2. The reaction solution was concentrated and filtered with a micromembrane filter. The filtrate was filtered with C 18 Column, separated by preparative HPLC, chromatographic conditions: 15% CH3CN + 1% AcOH aqueous solution; flow rate: 5 mL / min; detection by UV detector, compound W9 retention time t = 250 min, compound W10 retention time t = 300 min, to obtain two compounds W9 (16.5 mg, 11%) and W10 (43.3 mg, 29%), both as white solids.

[0246] The obtained product W9 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0247] 1H NMR (600MHz, DMSO-d6) δ12.01(s,1H),8.35(s,1H),7.98(d,J=7.9Hz,1H),7.37–7.23(m,5H),5.73(s,2H),5.61(d,J=5.1Hz,1H),5.39(d,J= 9.3Hz,1H),5.23(s,1H),5.07–4.96(m,2H),4.35(s,1H),3.90(d,J=9.8Hz,2H),3.72(s,1H),3.51(d,J=9.9Hz,2H),2.83(d,J=27.4Hz,3H). 13 C NMR (151MHz, DMSO-d6) δ170.14,169.51,168.56,159.08,156.48,156.20,150.24,137.49,137.40,128.88,128.25,128. 05,127.88,127.60,104.69,76.87,73.55,71.22,66.71,66.52,62.76,60.23,55.66,55.37,52.43,51.33,21.34,14.53. 19 F NMR(376MHz,DMSO-d6)δ-61.53.ESI-HRMS(m / z):calcd for C 25 H 28 F3N5O 12 :[M+Na] + 670.15788;found:670.15770.[a] D 20 +13.7 (c 0.10, DMF).

[0248] The obtained product W10 was measured on a nuclear magnetic resonance instrument, and the obtained nuclear magnetic resonance data were:

[0249] 1H NMR (600MHz, DMSO-d6) δ12.01(s,1H),8.35(s,1H),7.99(d,J=8.1Hz,1H),7. 37–7.23(m,5H),5.73(s,1H),5.59(d,J=5.0Hz,1H),5.39(d,J=9.3Hz,1H),5. 11(d,J=4.3Hz,1H),5.06–4.96(m,2H),4.29(s,1H),4.11(s,1H),3.94–3.85 (m,2H),3.73(s,1H),3.56(s,1H),3.53–3.47(m,2H),2.83(d,J=28.6Hz,3H). 13 C NMR (151MHz, DMSO-d6) δ170.45,169.56,168.65,159.08,156.52,156.21,150.28,137.51,137.39,128.88,128.78,128.26,128. 07,127.90,127.60,122.03,76.64,73.83,71.07,66.73,66.51,62.57,60.22,55.83,55.37,53.14,51.68,51.31,21.54,14.53. 19 F NMR(376MHz,CD3OD)δ-64.70.ESI-HRMS(m / z):calcd for C 25 H 28 F3N5O 12 :[M+Na] + 670.15788;found:670.15763.[a] D 20 +16.3(c0.10,DMF).

[0250] 5. Synthesize compound W11, the chemical reaction equation is:

[0251]

[0252] The specific synthesis process is:

[0253] Compound W9 (27.9 mg, 0.0431 mmol) was dissolved in EtOH (3 mL) and water (3 mL), and Pd / C (5.6 mg) was added. After reacting at room temperature for 30 minutes under hydrogen, the reaction was complete after TLC monitoring. Filter, concentrate, and use C 18The column was used for HPLC preparation. The chromatographic conditions were: 5% CH3CN + 1% AcOH aqueous solution; flow rate: 5 mL / min; detection by UV detector, retention time of compound W11 was t=15 min. After the eluate was concentrated under reduced pressure, pyrimidine nucleoside peptide compound W11 (11.2 mg, 86%) was obtained.

[0254] The obtained product W11 was measured on a nuclear magnetic resonance instrument, and the nuclear magnetic resonance data obtained were:

[0255] 1 H NMR(400MHz,D2O)δ8.32(d,J=8.7Hz,1H),5.74–5.60(m,1H),4.19(s,1H),4 .07(m,J=17.8,7.4Hz,4H),3.80(dd,J=9.3,4.4Hz,3H),3.00–2.86(m,3H). 13 C NMR(101MHz,D2O)δ176.66,172.95,172.79,169.34,160.96,157.18,150.84,143.06,82. 57,75.49,73.15,71.40,71.35,58.21,58.13,56.05,55.88,52.95,51.66,29.19,20.38. 19 F NMR(376MHz,D2O)δ-63.44.ESI-HRMS(m / z):calcd for C 17 H 22 F3N5O 10 :[M+K] + 552.09503; found:552.09502.[a] D 20 +38.2 (c 0.11, DMSO).

[0256] 6. Synthesize compound W12, the chemical reaction equation is:

[0257]

[0258] The specific synthesis process is:

[0259] Compound W10 (27.9 mg, 0.0431 mmol) was dissolved in EtOH (3 mL) and water (3 mL), and Pd / C (5.6 mg) was added. After reacting at room temperature for 30 minutes under hydrogen, the reaction was complete after TLC monitoring. Filter, concentrate, and use C 18The column was used for HPLC preparation. The chromatographic conditions were: 5% CH3CN + 1% AcOH aqueous solution; flow rate: 5 mL / min; detection by UV detector, retention time of compound W12 was t=15 min. After the eluate was concentrated under reduced pressure, pyrimidine nucleoside peptide compound W12 (30.90 mg, 90%) was obtained.

[0260] The obtained product W12 was measured on a nuclear magnetic resonance instrument, and the nuclear magnetic resonance data obtained were:

[0261] 1 H NMR (400MHz, D2O) δ8.37 (s, 1H), 5.64 (d, J = 4.1Hz, 1H), 4.45 (d, J = 5.0Hz, 1H), 4.03–3.85 (m, 4H), 3.81–3.68 (m, 4H), 2.70 (d, J = 4.0Hz, 3H). 13 C NMR(101MHz,D2O)δ181.42,173.86,171.34,171.23,166.50,166.39,161.52,151.24,143.27,123.76,120.84,105.85,8 2.62,77.86,73.50,73.39,71.95,71.82,61.45,61.29,55.77,55.68,53.32,53.26,49.52,49.41,32.86,32.84,23.23. 19 F NMR(376MHz,D2O)δ-63.36.ESI-HRMS(m / z):calcd for C 17 H 22 F3N5O 10 :[M+Na] + 536.12110; found:536.12119.[a] D 20 +55.6 (c 0.10, DMF).

[0262] Example 5 Activity Test

[0263] Compounds W1 to W12 were tested for their anti-plant pathogen activity, and the pathogens used included major plant disease pathogens such as Sclerotinia sclerotiorum, Gibberella fusca, and Pyricularia oryzae. Figure 1 As shown, it was found that this type of compound had a certain degree of inhibition rate against all pathogens used, especially the inhibition rate against Sclerotinia sclerotiorum was generally higher than that against wheat fusarium head blight and rice blast, indicating that this type of structural compound has good selectivity against Sclerotinia sclerotiorum. Among them, compound W8 showed good antibacterial activity, with an EC50 of 34.93 mg / L.

[0264] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A pyrimidine nucleoside peptide agricultural antibiotic, characterized in that: Its structural formula is any one of the following formula (I): In the formula, R 1 is any one of a hydrogen atom, an alkyl group, an aryl group, a heteroaryl group, an alkoxy group, an amino group, a substituted amino group, an ester group, an alkenyl group, an alkynyl group, a carboxyl group, a halogen group, a nitro group, a fluoroalkyl group, a cyano group, a sulfone group, an amide group, and a phosphate group; R 2 It is any one of an alkoxy group, a hydroxyl group, an amino group, a substituted amine group, and a cyclic amine group; R 3 , R 4 are any one of hydrogen atom, aromatic acyl group, heteroaromatic acyl group, fatty acyl group and sulfonyl group; or, R 3 O.R 4 O is substituted by any one of an azide group, a substituted amino group, a thiol group, an alkyl sulfide, an aryl sulfide, an alkyl group, an alkenyl group, and an alkynyl group; R 5 is any one of a hydroxyl group, an ester group, an alkoxy group, an alkyl group, an aryl group, a heteroaryl group, an azido group, an amino group, a substituted amino group, a dialkylamino group, and a cyclic amino group; R 6 is any one of a hydrogen atom, a benzyloxycarbonyl group, a tert-butyloxycarbonyl group, an alkyl group, an aromatic acyl group, a fatty acyl group, and a sulfonyl group; n is any natural number from 0 to 10.

2. A pyrimidine nucleoside peptide agricultural antibiotic according to claim 1, characterized in that: When the R 1 When R is any one of a hydrogen atom, an alkyl group, an aryl group, a heteroaryl group, an alkoxy group, an amino group, a substituted amino group, an ester group, an alkenyl group, an alkynyl group, a carboxyl group, a halogen group, a nitro group, a fluoroalkyl group, a cyano group, a sulfone group, an amide group, and a phosphate group, 1 Can be represented by one or more independent R X Substituents are substituted; When the R 2 When R is any one of an alkoxy group, a hydroxyl group, an amino group, a substituted amine group, and a cyclic amine group; 2 Can be represented by one or more independent R X Substituents are substituted; When the R 3 , R 4 When R 3 , R 4 Can be represented by one or more independent R X Substituents are substituted; Or, when the R 3 O.R 4 When O is independently any one of an azido group, a substituted amino group, a mercapto group, an alkyl sulfide, an aryl sulfide, an alkyl group, an alkenyl group, and an alkynyl group, R 3 O.R 4 O can be represented by one or more independent R X Substituents are substituted; When the R 5 When R is any one of a hydroxyl group, an ester group, an alkoxy group, an alkyl group, an aryl group, a heteroaryl group, an azido group, an amino group, a substituted amino group, a dialkylamino group, and a cyclic amino group, 5 Can be represented by one or more independent R X Substituents are substituted; When the R 6 When R is any one of a hydrogen atom, a benzyloxycarbonyl group, a tert-butyloxycarbonyl group, an alkyl group, an aromatic acyl group, an aliphatic acyl group, and a sulfonyl group, 6 Can be represented by one or more independent R X Substituents are substituted; The R X It is any one of a hydrogen atom, a halogen, an alkyl group, a fluoroalkyl group, an alkenyl group, an alkynyl group, a nitro group, a cyano group, a cycloalkyl group, an aryl group, an amino group, an alkoxy group, a substituted amino group, an amide group, a hydroxyl group, a sulfonamide group, a diazo group, an azido group, an alkyl acyl group, and an aryl acyl group.

3. A pyrimidine nucleoside peptide agricultural antibiotic according to claim 2, characterized in that: When the R X When R is any one of a hydrogen atom, a halogen, an alkyl group, a fluoroalkyl group, an alkenyl group, an alkynyl group, a nitro group, a cyano group, a cycloalkyl group, an aryl group, an amino group, an alkoxy group, a substituted amino group, an amide group, a hydroxyl group, a sulfonamide group, a diazo group, an azido group, an alkyl acyl group, and an aryl acyl group, X It may be substituted by one or more hydrogen atoms, halogen, alkyl, fluoroalkyl, alkenyl, alkynyl, nitro, cyano, cycloalkyl, aryl, amino, alkoxy, substituted amino, amide, hydroxyl, sulfonamide, diazo, azido, alkyl acyl, aryl acyl, respectively.

4. A method for synthesizing the pyrimidine nucleoside peptide agricultural antibiotic according to any one of claims 1 to 3, characterized in that: Compound 7 or compound 8 and Pd / C are added to a mixed solvent of EtOH and water, and the reaction is completed at room temperature in the presence of hydrogen, and Pd / C is removed by filtration and concentrated to obtain a pyrimidine nucleoside peptide compound; The compound 7 or compound 8 is synthesized by compound 6, and the synthesis process is as follows: dissolving compound 6 in CH3OH, adding an aqueous solution of LiOH at 0°C for hydrolysis, the reaction is complete, adjusting the pH to 2 with glacial acetic acid, retaining the aqueous phase, concentrating and filtering the aqueous phase, and separating and purifying the filtrate by HPLC to obtain compound 7 or compound 8; the equivalent ratio of LiOH to compound 6 is 0.01:1 to 50:1; the structural formula of compound 6 is:

5. The synthesis method according to claim 4, characterized in that: The synthesis method of compound 6 is as follows: using compound 4 and compound 5 as raw materials, HATU as a condensation agent, and 2,6-dimethylpyridine as a base, reacting fully under argon protection, extracting, washing, drying, filtering and concentrating, and separating by silica gel column chromatography to obtain compound 6; The equivalent ratio of compound 5 to compound 4 is 0.01:1 to 50:1, the equivalent ratio of HATU to compound 4 is 0.01:1 to 50:1, and the equivalent ratio of 2,6-lutidine to compound 4 is 0.01:1 to 50:1; The structural formulas of the compounds 4 and 5 are:

6. The synthesis method according to claim 5, characterized in that: The synthesis process of compound 4 is as follows: 1) Compound 3 is dissolved in a mixed solvent of CH3CN and H2O, TEMPO is added, and then iodobenzene acetate is added. After the reaction is complete, anhydrous CH3OH is added to quench the reaction, and the crude carboxylic acid compound is concentrated to obtain a crude carboxylic acid compound, which can be subjected to the next step of reaction after pulping; the equivalent ratio of TEMPO to compound 3 is 0.001:1 to 10:1, and the equivalent ratio of iodobenzene acetate to compound 3 is 0.001:1 to 100:1; 2) the carboxylic acid compound is dissolved in CH3OH, H2SO4 is added, refluxed, concentrated after the reaction is complete, the reaction solution is extracted with ethyl acetate, the organic phase is retained, dried, filtered, and concentrated under reduced pressure to obtain a crude methyl esterification product; the equivalent ratio of H2SO4 to the carboxylic acid compound is 0.001:1 to 50:1; 3) reacting the methyl ester compound with SnCl2·2H2O, p-methylthiophenol, CHCl3 and Et3N, and concentrating under reduced pressure until the reaction is complete, adding ethyl acetate to dissolve, filtering to remove the insoluble SnCl2·2H2O, concentrating the filtrate, and separating and purifying by silica gel column chromatography to obtain compound 4; The equivalent ratio of SnCl2·2H2O to the methyl ester compound is 0.01:1 to 50:1, the equivalent ratio of p-methylthiophenol to the methyl ester compound is 0.01:1 to 100:1, and the equivalent ratio of Et3N to the methyl ester compound is 0.01:1 to 300:1; The structural formula of the compound 3 is:

7. The synthesis method according to claim 6, characterized in that: The synthesis process of the compound 3 is as follows: using 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative as substrate, adding uracil or uracil with different substituents, adding catalyst, BSA and acetonitrile, and performing condensation reaction under heating reflux conditions to obtain nucleoside products; The catalyst is any one of BF3·Et2O, TMSOTf, TBSOTf, TESOTf, metal halide, metal sulfonate, perchloric acid and its metal salt, triarylborane, organic phosphoric acid, organic thiourea, and organic carbonyl acid.

8. The synthesis method according to claim 7, characterized in that: The equivalent ratio of the catalyst to the 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative is 1:0.01-1:100, the equivalent ratio of uracil or substituted uracil to the 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative is 1:0.01-1:100, the equivalent ratio of BSA to the 1-O-acetyl-4-azido-4-deoxy-D-glucose derivative is 1:0.01-1:100, the equivalent ratio of the nucleoside product to sodium hydroxide is 1:0.01-1:100, the reaction concentration is 0.01-100 mol / L, and the reaction temperature is 60-200°C.

9. An agricultural antibiotic composition, characterized in that: The invention comprises the pyrimidine nucleoside peptide agricultural antibiotic as described in any one of claims 1 to 3 or the pyrimidine nucleoside peptide agricultural antibiotic synthesized by the synthesis method as described in any one of claims 4 to 8, or its stereoisomers, tautomers or salts thereof, or its prodrug molecules and a pesticide acceptable carrier.

10. Use of a pyrimidine nucleoside peptide agricultural antibiotic as described in any one of claims 1 to 3 or a pyrimidine nucleoside peptide agricultural antibiotic synthesized by the synthesis method as described in any one of claims 4 to 8, or its stereoisomers, tautomers or salts, or its prodrug molecules, or the agricultural antibiotic composition as described in claim 9 in the prevention and control of plant diseases, wherein the plant diseases include rapeseed sclerotinia rot, rice blast, and wheat fusarium head blight.

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