Preparation of 13 sugar and substructure 9 sugar of angelica polysaccharide aps-1ii with anti-leukemia activity

Abstract

The application provides a method for constructing 1,2-cis fucose glycoside bond with high stereoselectivity and application in preparation of 13-sugar repeating units and 9-sugar substructures of angelica polysaccharide APS-1II, and evaluation of anti-leukemia activity of the 13-sugar and 9-sugar, and belongs to the technical field of polysaccharide preparation; the application realizes a promoter composition of 1,2-cis fucose glycosylation reaction with high stereoselectivity, which comprises iodotrimethylsilane (TMSI) and triphenyl phosphine oxide (Ph3PO), and the donor is fucosyl N-phenyl trifluoroacetyl imidate (PTFAI) with acyl substitution at O-3 and / or O-4 positions. The 13-sugar repeating units and 9-sugar substructures of angelica polysaccharide APS-1II provided by the application have significant inhibitory effect on human leukemia K562 cells and mouse leukemia L1210 cells.

Description

Technical Field

[0001] This invention belongs to the field of polysaccharide preparation technology, specifically relating to a method for highly stereoselective 1,2-cis-fucosylation reaction and its application in the preparation of the 13-saccharide repeating unit and substructure 9-saccharide of Angelica sinensis polysaccharide APS-1 II with anti-leukemia activity. Background Technology

[0002] Angelica sinensis is a traditional Chinese medicine with remarkable effects on gynecological diseases in women and has been widely used in the treatment of various ailments. Studies have shown that angelica polysaccharides are the main active substances in angelica, possessing beneficial effects such as enhancing immunity, anti-oxidation, anti-tumor activity, and anti-radiation properties.

[0003] In 2018, Professor Cao Wei of Northwest A&F University isolated and extracted a novel APS-1 II polysaccharide with anti-leukemia activity from the root of Angelica sinensis. Structurally, APS-1 II polysaccharide has the following characteristics: 1) it uses a highly branched 13-saccharide as the repeating unit, while also containing two 3,5-branched arabinofuranose units; 2) it contains a challenging α-glucose-(1→3) glycosidic bond; 3) it contains three 1,2-cis-fucoside bonds, which also make it difficult to achieve high stereoselectivity. Bioactivity studies showed that APS-1 II polysaccharide can inhibit the activity of human leukemia K562 cells and mouse leukemia L1210 cells. Furthermore, it can significantly prolong the lifespan of mice infected with the L1210 leukemia strain.

[0004] However, there are currently no reports on the synthesis of the 13-saccharide repeating unit and its substructure of APS-1 II polysaccharide by methods other than extraction. Summary of the Invention

[0005] In view of this, the present invention aims to provide a method for highly stereoselective 1,2-cis-fucosylation reaction and its application in the preparation of the 13-saccharide repeating unit and substructure 9 sugar of Angelica polysaccharide APS-1 II. The promoter composition and fucose PTFAI donor provided by the present invention can regulate the stereoselectivity of the glycosylation reaction product. The highly stereoselective 1,2-cis-fucosylation reaction method provided by the present invention can achieve highly stereoselective control of the 1,2-cis-fucosylation bond. The 13-saccharide repeating unit and substructure 9 sugar of Angelica polysaccharide APS-1 II provided by the present invention have a significant inhibitory effect on the activity of human leukemia K562 cells and mouse leukemia L1210 cells.

[0006] This invention provides a promoter composition for regulating the stereoselectivity of glycosylation reactions, comprising triiodosilane (TMSI) and triphenylphosphine oxide (Ph3PO); the molar ratio of triiodosilane to triphenylphosphine oxide is 1:(5-10). This invention also provides a fucose PTFAI donor for highly stereoselective 1,2-cis-fucosylation reactions, wherein the O-3 and / or O-4 positions of the fucose PTFAI donor are substituted with acyl groups. The promoter composition and fucose PTFAI donor provided by this invention can regulate the stereoselectivity of the glycosylation reaction product, giving it high stereoselectivity.

[0007] This invention provides a method for highly stereoselective 1,2-cis-fucoside glycosylation, which can achieve highly stereoselective control of 1,2-cis-glycosidic bonds, with a stereoselectivity of α / β of glycosylation product > 20:1 and a yield of 80-99%; at the same time, it can construct three consecutive 1,2-cis-fucoside bonds with high stereoselectivity.

[0008] This invention provides a 13-saccharide repeating unit of Angelica polysaccharide APS-1 II and its substructure 9-saccharide, which has a significant inhibitory effect on the activity of human leukemia K562 cells and mouse leukemia L1210 cells. Detailed Implementation

[0009] The present invention provides a promoter composition for highly stereoselective 1,2-cis-fucosylation reaction, comprising triiodosilane and triphenylphosphine oxide.

[0010] In this invention, the molar ratio of triiodotrimethylsilane to triphenylphosphine oxide is 1:(5-10). In the embodiments of this invention, it can specifically be 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

[0011] This invention provides a fucose PTFAI donor for highly stereoselective 1,2-cis-fucosylation, wherein the O-3 and / or O-4 positions of the fucose PTFAI donor are substituted with an acyl group. In this invention, the acyl group preferably includes -Bz or -Lev.

[0012] This invention provides the application of the promoter composition described above or the fucose PTFAI donor described above in glycosylation reactions. The promoter composition and fucose PTFAI donor provided by this invention can adjust the stereoselectivity of the glycosylation reaction products, giving them high stereoselectivity. Unless otherwise specified, the materials and equipment used in this invention are commercially available.

[0013] This invention provides a method for highly stereoselective 1,2-cis-fucosylation reaction, comprising the following steps:

[0014] A fucose PTFAI donor, an alcohol acceptor, a promoter composition, a dehydrating agent, and a polar organic solvent are mixed and subjected to a glycosylation reaction to obtain a glycosylated product having a 1,2-cis-fucose glycosidic bond; the stereoselectivity of the glycosylated product is α / β > 20:1.

[0015] In this invention, the fucose PTFAI donor preferably includes a monosaccharide PTFAI donor or a polysaccharide PTFAI donor; the monosaccharide PTFAI donor preferably has the structure shown in Formula 21; in Formula 21, R 2 For -Bn, -Me, or -Nap, R 3 For -Bz, -Ac, or -Lev; R 4 The polysaccharide PTFAI donor is either -Bz or -Ac; preferably, the polysaccharide PTFAI donor has the structure shown in Formula 9.

[0016]

[0017] In this invention, the alcohol receptor preferably has the structure shown in Formula 22, Formula 24, Formula 29 or Formula 10; HO(CH2)5NBnCbz Formula 22; H-OR Formula 24; in Formula 24, -OR has the structure shown in Formula 24d, Formula 24m or Formula 24v;

[0018]

[0019] In this invention, the compounds with the structure shown in Formula 22 are referenced to: Castelli, Riccardo; Overkleeft, Herman S.; van der Marel, Gijsbert A.; Codée, Jeroen DC. (2013). 2,2-Dimethyl-4-(4-methoxy-phenoxy)

[0020] butanoate and 2,2-Dimethyl-4-azido Butanoate: Two New Pivaloate-ester-like Protecting Groups. Organic Letters, 15(9), 2270–2273. Preparation.

[0021] Compounds with the structure shown in Formula 24 are referenced in: Penghua Li, Hanyingzi Fan, Qiang Tan, Guozhi Xiao; Highly Stereoselective Assembly of 1,2-cis-Arap Linkages; Organic Letters 2023 25(16), 2788-2792.

[0022] In this invention, when the monosaccharide PTFAI donor has the structure shown in Formula 21, the alcohol acceptor has the structure shown in Formula 22, Formula 24 or Formula 29;

[0023] When the polysaccharide PTFAI donor has the structure shown in Formula 9, the alcohol acceptor has the structure shown in Formula 10.

[0024] In this invention, formula 21 preferably has the structure shown in formula 21d:

[0025]

[0026] The present invention does not have any special requirements for the preparation of compounds with the structure shown in Formula 21d; they can be obtained by means well known in the art.

[0027] In this invention, the accelerator composition is the accelerator composition described in the above-described scheme; the accelerator composition includes trimethylsilimidazole and triphenylphosphine oxide. In this invention, the polar organic solvent preferably includes dichloromethane (DCM); the dehydrating agent preferably includes molecular sieve (MS).

[0028] In this invention, the mixing of the fucose PTFAI donor, alcohol acceptor, promoter composition, dehydrating agent, and polar organic solvent preferably includes: first, a first mixing of the triphenylphosphine oxide in the fucose PTFAI donor, alcohol acceptor, and promoter composition with the polar organic solvent; then, a second mixing is performed by adding the dehydrating agent under inert gas protection; and finally, triiodomethylsilane is added dropwise. In this invention, the addition of the dehydrating agent, the dropwise addition of triiodomethylsilane, and the glycosylation reaction steps all need to be carried out under inert gas protection.

[0029] In this invention, prior to the first mixing, it is preferable to further dehydrate the fucose PTFAI donor and the alcohol acceptor. This invention does not have specific requirements for the method of dehydration; any method well-known in the art can be used. In embodiments of this invention, it is preferable to azeotropically react the fucose PTFAI donor and the alcohol acceptor with toluene; the azeotropic reaction is preferably repeated three times.

[0030] In this invention, prior to the first mixing, it is preferable to further remove water and impurities from the polar organic solvent. Prior to the first mixing, it is also preferable to activate the dehydrating agent. This invention does not have particular requirements regarding the activation method of the dehydrating agent; any method well-known in the art can be used. The inert gas preferably includes argon.

[0031] In this invention, the second mixing time is preferably 10 to 15 minutes, and in specific embodiments of this invention, it can be 10 minutes, 12 minutes, or 15 minutes.

[0032] In this invention, the molar ratio of the fucose PTFAI donor to the alcohol acceptor is preferably (1-1.6):(1-1.6), and in the embodiments of this invention, it can specifically be 1:1, 1:1.5, 1:1.6, 1.45:1, 1.49:1, 1.5:1, or 1.6:1; the molar ratio of iodotrimethylsilane in the fucose PTFAI donor and promoter composition is preferably (1-1.6):1, and in the embodiments of this invention, it can specifically be 1:1, 1.45:1, 1.49:1, 1.5:1, or 1.6:1; the concentration of the fucose PTFAI donor in the polar organic solvent is preferably 0.1-0.2 mol / L, and in the embodiments of this invention, it can specifically be 0.1 mol / L, 0.15 mol / L, or 0.2 mol / L.

[0033] In this invention, the glycosylation reaction time is preferably 24 hours to 5 days. In specific embodiments of this invention, it can be 24 hours, 38 hours, 48 ​​hours, 72 hours, 80 hours, 96 hours, or 4.5 days. The glycosylation reaction time is preferably calculated from the completion of the addition of the trimethyl iodosilane. In this invention, the glycosylation reaction temperature is preferably room temperature.

[0034] After the glycosylation reaction is completed, the present invention preferably performs post-treatment on the resulting reaction solution. The post-treatment preferably includes diluting the resulting reaction solution with ethyl acetate (EA) and quenching the reaction with saturated Na₂S₂O₃ solution. The organic phase is washed with water and saturated brine, dried over anhydrous Na₂SO₄, filtered and concentrated, and finally purified by silica gel column chromatography. The present invention does not have particular requirements for the specific method of the post-treatment; methods well known in the art can be used.

[0035] This invention provides an Angelica sinensis polysaccharide APS-1 II 13-sugar repeating unit and its substructure 9-sugar. The Angelica sinensis polysaccharide APS-1 II 13-sugar repeating unit has the structure shown in Formula 1, and the substructure 9-sugar has the structure shown in Formula 2.

[0036]

[0037] In this invention, the method for preparing the substructure 9 sugar includes the following steps:

[0038] Fucose PTFAI donor 12, fucose acceptor 11, first promoter composition, first polar organic solvent and first dehydrating agent are mixed and subjected to a first glycosylation reaction to obtain the first glycosylation product disaccharide 28; the first glycosylation product disaccharide 28 is deprotected by the Lev protecting group to obtain disaccharide acceptor 29.

[0039] The fucose PTFAI donor 12 has the structure shown in Formula 12; the fucose acceptor 11 has the structure shown in Formula 240; the first glycosylation product disaccharide 28 has the structure shown in Formula 28; the disaccharide acceptor 29 has the structure shown in Formula 29; the first promoter composition is the promoter composition described in the above scheme; the promoter composition includes triiodosilane and triphenylphosphine oxide.

[0040]

[0041] The present invention does not have any special requirements for the preparation of compounds with the structure shown in Formula 12; they can be obtained by means well known in the art.

[0042] In this invention, the conditions for the first glycosylation reaction and the ratio of each raw material are the same as those for the glycosylation reaction described above, and will not be repeated here.

[0043] In this invention, removing the Lev protecting group from the first glycosylated product disaccharide 28 preferably includes the following steps: mixing the first glycosylated product disaccharide 28, a pyridine-acetic acid (pyridine / AcOH) mixed solvent, and hydrazine hydrate (NH2NH2·H2O) to remove the Lev protecting group, thereby obtaining the disaccharide acceptor 29.

[0044] In this invention, the volume ratio of pyridine to acetic acid in the pyridine-acetic acid mixed solvent is preferably 3:2; the molar amount of the first glycosylation product disaccharide 28 and the volume ratio of the pyridine-acetic acid mixed solvent are preferably 0.48 mmol: 4.7 mL; the molar ratio of the first glycosylation product disaccharide 28 to hydrazine hydrate is preferably 1:(2.5-3.5), and in the embodiments of this invention, it can specifically be 1:2.5, 1:2.98, 1:3 or 1:3.5.

[0045] After removing the Lev protecting group, the present invention preferably performs post-treatment on the resulting reaction solution. The post-treatment preferably includes quenching and concentrating the resulting reaction solution with acetone, diluting the mixture with ethyl acetate, washing with 4M HCl, saturated NaHCO3 solution, and saturated brine, drying with anhydrous Na2SO4, filtering, concentrating, and finally purifying by column chromatography. The present invention does not have specific requirements for the specific method of the post-treatment; methods well known in the art can be used.

[0046] After obtaining the disaccharide receptor 29, the present invention mixes the fucose PTFAI donor 21d with the disaccharide receptor 29, the second promoter composition, the second polar organic solvent and the second dehydrating agent to carry out a second glycosylation reaction to obtain the trisaccharide 30;

[0047] The fucose PTFAI donor 21d has the structure shown in Formula 21d; the trisaccharide 30 has the structure shown in Formula 30;

[0048] The second accelerator composition is the accelerator composition described in the above scheme; the accelerator composition includes triiodosilane and triphenylphosphine oxide;

[0049]

[0050] In this invention, the conditions for the second glycosylation reaction and the ratio of each raw material are the same as those for the glycosylation reaction described above, and will not be repeated here.

[0051] After obtaining the trisaccharide 30, the present invention selectively removes the protecting group STol from the trisaccharide 30 to obtain a trisaccharide intermediate.

[0052] In this invention, selectively removing the protecting group STol from the trisaccharide 30 preferably includes the following steps:

[0053] The trisaccharide 30 was dissolved in an aqueous acetone solution, and the resulting mixed solution was cooled. Trichloroisocyanuric acid (TCCA) was added to remove the STol protecting group, yielding the trisaccharide intermediate.

[0054] In this invention, the volume ratio of acetone to water in the acetone aqueous solution is preferably 4:1; the molar amount of the trisaccharide 30 and the volume ratio of the acetone aqueous solution are preferably 0.19 mmol: 4.6 mL; the molar ratio of the trisaccharide 30 and trichloroisocyanuric acid is preferably 1:1; the cooling is preferably to 0°C; the removal time of the STol protecting group is preferably 10 h, and the temperature is preferably room temperature.

[0055] After removing the STol protecting group, the present invention preferably performs post-treatment on the resulting reaction solution; the post-treatment preferably includes: dilution with ethyl acetate, washing with saturated NaHCO3 solution, water and saturated brine, drying with anhydrous Na2SO4, filtration, concentration, and finally purification by column chromatography. The present invention does not have particular requirements on the specific method of the post-treatment; methods well known in the art can be used.

[0056] After obtaining the trisaccharide intermediate, the present invention mixes the trisaccharide intermediate, 2,2,2-trifluoro-N-phenylacetimidoyl chloride, a third polar organic solvent, and a first carbonate to carry out a first substitution reaction to obtain a trisaccharide PTFAI donor 9 with a PTFAI protecting group; the trisaccharide PTFAI donor 9 has the structure shown in Formula 9;

[0057]

[0058] In this invention, the third polar organic solvent preferably includes acetone; the first carbonate preferably includes potassium carbonate. In this invention, the carbonate acts as an acid-binding agent to neutralize the HCl produced in the reaction.

[0059] In this invention, the molar ratio of the trisaccharide intermediate to 2,2,2-trifluoro-N-phenyliminoacetyl chloride is preferably 1:(1.8-2), and in the embodiments of this invention, it can specifically be 1:1.8, 1:1.86, or 1:2; the molar ratio of the trisaccharide intermediate to the volume ratio of the third polar organic solvent is preferably 0.07 mmol:0.7 mL; and the molar ratio of the 2,2,2-trifluoro-N-phenyliminoacetyl chloride to the first carbonate is preferably 1:1.

[0060] In this invention, the temperature of the first substitution reaction is preferably room temperature, and the time is preferably 10 hours.

[0061] After the first substitution reaction is completed, the present invention preferably performs post-treatment on the resulting reaction solution; the post-treatment preferably includes concentration and column chromatography separation and purification. The present invention does not have any particular requirements on the specific method of the post-treatment, and any method well known in the art can be used.

[0062] After obtaining the trisaccharide PTFAI donor 9, the present invention mixes the trisaccharide PTFAI donor 9, the hexasaccharide acceptor 10, the third promoter composition, the fourth polar organic solvent and the third dehydrating agent, and performs a third glycosylation reaction to obtain the nonaconose 31; the hexasaccharide 10 has the structure shown in Formula 10; the nonaconose 31 has the structure shown in Formula 31;

[0063] The third accelerator composition is the accelerator composition described in the above scheme; the accelerator composition includes triiodosilane and triphenylphosphine oxide;

[0064]

[0065] In this invention, the conditions for the third glycosylation reaction and the ratio of each raw material are the same as those for the glycosylation reaction described above, and will not be repeated here.

[0066] In this invention, the preparation of the hexasaccharide receptor 10 preferably includes the following steps:

[0067] The arabinose PTFAI donor 18, glucose PVB acceptor 19, first activator, fourth dehydrating agent, and fifth polar organic solvent are mixed and subjected to a fourth glycosylation reaction to obtain a reaction solution containing α-Araf-(1→6)-Glc disaccharide; the reaction solution containing α-Araf-(1→6)-Glc disaccharide, arabinose acceptor 20, second activator, and N,N-dimethylformamide (DMF) are mixed and subjected to a fifth glycosylation reaction to obtain trisaccharide 26;

[0068] The arabinose PTFAI donor 18 has the structure shown in Formula 18; the glucose PVB receptor 19 has the structure shown in Formula 19; the arabinose receptor 20 has the structure shown in Formula 20; and the trisaccharide 26 has the structure shown in Formula 26.

[0069]

[0070] The present invention does not have any special requirements for the preparation of the compounds with the structures shown in Formulas 18 to 20; they can be obtained by means well known in the art.

[0071] In this invention, the first activator is preferably trimethylsilyl trifluoromethanesulfonate (TMSOTf); the fourth dehydrating agent preferably includes a molecular sieve; and the fifth polar organic solvent preferably includes dichloromethane.

[0072] In this invention, the molar ratio of arabinose PTFAI donor 18 to glucose PVB acceptor 19 is preferably 1.2:1; the molar ratio of arabinose PTFAI donor 18 to the first activator is preferably (3.5-4):1, and in embodiments of this invention, it can specifically be 3.5:1, 3.8:1, or 4:1. In this invention, the mass ratio of arabinose PTFAI donor 18 to the fourth dehydrating agent is preferably 1:(1.5-2.5), and in embodiments of this invention, it can specifically be 1:1.5, 1:2, 1:2.1, or 1:2.5. In this invention, the molar amount of arabinose PTFAI donor 18 and the volume ratio of the fifth polar organic solvent are preferably 0.42 mmol:3.2 mL.

[0073] In this invention, the mixing of arabinose PTFAI donor 18, glucose PVB acceptor 19, first activator, fourth dehydrating agent and fifth polar organic solvent preferably includes: mixing and stirring arabinose PTFAI donor 18, glucose PVB acceptor 19, fourth dehydrating agent and fifth polar organic solvent, then cooling, and then adding the first activator dropwise.

[0074] In this invention, the mixing and stirring time is preferably 15 minutes; the cooling is preferably to -40°C. In this invention, the fourth glycosylation reaction time is preferably 2 hours, and the temperature is preferably -40°C; the time of the fourth glycosylation reaction is preferably calculated from the completion of the addition of the first activator.

[0075] After obtaining the reaction solution containing α-Araf-(1→6)-Glc disaccharide, the present invention preferably does not perform post-processing. Then, the reaction solution containing α-Araf-(1→6)-Glc disaccharide, arabinose acceptor 20, second activator, and N,N-dimethylformamide are mixed and subjected to the fifth glycosylation reaction to obtain trisaccharide 26.

[0076] In this invention, the mixing of the reaction solution containing α-Araf-(1→6)-Glc disaccharide, arabinose acceptor 20, second activator, and N,N-dimethylformamide preferably includes: heating the reaction solution containing α-Araf-(1→6)-Glc disaccharide, then adding N,N-dimethylformamide, performing a first stirring, then adding the second activator, performing a second stirring, then adding the arabinose acceptor 20, heating the resulting reaction solution to room temperature, and performing a fifth glycosylation reaction.

[0077] In this invention, the heating is preferably carried out to 0°C; the temperature of the first stirring is preferably 0°C, and the time is preferably 15 min; the time of the second stirring is preferably 1.5 h. In this invention, the second activator is preferably N-iodosuccinimide (NIS) and trimethyl trifluoromethanesulfonate (TMSOTf); the molar ratio of N-iodosuccinimide and trimethyl trifluoromethanesulfonate is preferably 1:1. In this invention, the reaction of NIS with TMSOTf releases positively charged iodide ions to activate PVB groups.

[0078] In this invention, the arabinose receptor 20 is preferably added in the form of an arabinose receptor 20 solution; in this invention, the arabinose receptor 20 solution is preferably obtained by dissolving the arabinose receptor 20 in dichloromethane; the concentration of the arabinose receptor 20 solution is preferably 0.29 mol / L.

[0079] In this invention, the molar ratio of the arabinose receptor 20 (referring to pure arabinose receptor 20) and the second activator is preferably 1:(2.5-3.5), and in the embodiments of this invention, it can specifically be 1:2.5, 1:3 or 1:3.5; the molar ratio of the arabinose receptor 20 and N,N-dimethylformamide is preferably 1:(15.5-16.5), and in the embodiments of this invention, it can specifically be 1:15.5, 1:16 or 1:16.5.

[0080] The preferred temperature for the fifth glycosylation reaction is room temperature, and the preferred time is 10 hours.

[0081] After the fifth glycosylation reaction is completed, the present invention preferably performs post-treatment on the resulting reaction solution; the post-treatment preferably includes: quenching the reaction with triethylamine (Et3N), filtration, concentration, and column chromatography separation and purification.

[0082] After obtaining the trisaccharide 26, the present invention removes the protecting group MP of the trisaccharide 26 to obtain a hemiacetal intermediate.

[0083] In this invention, the removal of the protecting group MP from the trisaccharide 26 preferably includes the following steps: dissolving the trisaccharide 26 in a toluene-acetonitrile aqueous solution (Toluene / MeCN / H2O), cooling, adding cerium ammonium nitrate (CAN), and removing the protecting group MP.

[0084] In this invention, the preferred volume ratio of toluene, acetonitrile, and water in the toluene-acetonitrile aqueous solution is 3:5:3; the preferred molar ratio of trisaccharide 26 to the toluene-acetonitrile aqueous solution is 1.85 mmol:37 mL; the preferred molar ratio of trisaccharide 26 to cerium ammonium nitrate is 1:(2-2.5), and in embodiments of this invention, it can specifically be 1:2, 1:2.2, 1:2.28, or 1:2.5. In this invention, the preferred removal time for the protecting group MP is 2 hours, and the preferred temperature is 0°C.

[0085] After the removal of the protecting group MP, the reaction mixture is preferably quenched with saturated NaHCO3 solution, extracted three times with DCM, washed with saturated brine, dried with Na2SO4, filtered, concentrated, and finally purified by column chromatography.

[0086] After obtaining the hemiacetal intermediate, the present invention mixes the hemiacetal intermediate, a sixth polar organic solvent, 2,2,2-trifluoro-N-phenyliminoacetyl chloride, and a second carbonate to carry out a second substitution reaction to obtain a trisaccharide PTFAI donor 17; the trisaccharide PTFAI donor 17 has the structure shown in Formula 17.

[0087]

[0088] In this invention, the sixth polar organic solvent preferably includes acetone; the second carbonate preferably includes potassium carbonate.

[0089] In this invention, the molar ratio of the hemiacetal intermediate to 2,2,2-trifluoro-N-phenyliminoacetyl chloride is preferably 1:(1.3-1.5), and in the embodiments of this invention, it can specifically be 1:1.3, 1:1.35, or 1:5; the molar ratio of the hemiacetal intermediate to the volume ratio of the sixth polar organic solvent is preferably 2.04 mmol:15.3 mL; and the molar ratio of the 2,2,2-trifluoro-N-phenyliminoacetyl chloride to the second carbonate is preferably 1:1.

[0090] In this invention, the temperature of the second substitution reaction is preferably room temperature, and the time is preferably 10 hours.

[0091] After the first substitution reaction is completed, the present invention preferably filters and concentrates the reaction mixture, and then separates and purifies it by column chromatography.

[0092] After obtaining the trisaccharide PTFAI donor 17, the present invention mixes the trisaccharide PTFAI donor 17, arabinose ABz acceptor 14, fifth dehydrating agent, seventh polar organic solvent and third activator, and performs a sixth glycosylation reaction to obtain a reaction solution containing a tetrasaccharide ABz intermediate.

[0093] In this invention, the third activator is preferably trimethylsilyl trifluoromethanesulfonate; the fifth dehydrating agent preferably includes a molecular sieve; and the seventh polar organic solvent preferably includes dichloromethane.

[0094] In this invention, the molar ratio of the trisaccharide PTFAI donor 17 to the arabinose ABz acceptor 14 is preferably 1.1:1; the molar ratio of the trisaccharide PTFAI donor 17 to the third activator is preferably (4-5):1, and in embodiments of this invention, it can specifically be 4:1, 4.5:1, or 5:1. In this invention, the mass ratio of the trisaccharide PTFAI donor 17 to the fifth dehydrating agent is preferably 1:(3-3.5), and in embodiments of this invention, it can specifically be 1:3, 1:3.2, or 1:3.5. In this invention, the volume ratio of the trisaccharide PTFAI donor 17 to the seventh polar organic solvent is preferably 0.09 mmol:0.4 mL.

[0095] In this invention, the mixing of the trisaccharide PTFAI donor 17, arabinose ABz acceptor 14, fifth dehydrating agent, seventh polar organic solvent and third activator preferably includes: mixing and stirring the trisaccharide PTFAI donor 17, arabinose ABz acceptor 14, fifth dehydrating agent and seventh polar organic solvent, then cooling, and then adding the third activator dropwise.

[0096] In this invention, the mixing and stirring time is preferably 15 minutes; the cooling is preferably to -40°C. In this invention, the time for the sixth glycosylation reaction is preferably 2 hours, and the temperature is preferably -40°C; the time for the sixth glycosylation reaction is preferably calculated from the completion of the addition of the third activator.

[0097] After obtaining the reaction solution containing the tetrasaccharide ABz intermediate, the present invention preferably does not perform post-processing, and then mixes the reaction solution containing the tetrasaccharide ABz intermediate, arabinose PVB acceptor 15 and catalyst to carry out the seventh glycosylation reaction to obtain the reaction solution containing the pentasaccharide intermediate.

[0098] In this invention, mixing the reaction solution containing the tetrasaccharide ABz intermediate, the arabinose PVB acceptor 15, and the catalyst preferably includes: heating the reaction solution containing the tetrasaccharide ABz intermediate to room temperature, and then adding the arabinose PVB acceptor 15 and the catalyst.

[0099] In this invention, the arabinose PVB receptor 15 is preferably added in the form of an arabinose PVB receptor 15 solution; the arabinose PVB receptor 15 solution is preferably obtained by dissolving the arabinose PVB receptor 15 in dichloromethane; the concentration of the arabinose PVB receptor 15 solution is preferably 0.35 mol / L.

[0100] In this invention, the catalyst is preferably added in the form of a catalyst solution; the catalyst solution is preferably obtained by dissolving the catalyst in dichloromethane; the concentration of the catalyst solution is preferably 0.12 mol / L. In this invention, the molar ratio of arabinose PVB acceptor 15 (referring to pure arabinose PVB acceptor 15) to catalyst (referring to pure catalyst) is preferably 7:1.2.

[0101] In this invention, the temperature of the seventh glycosylation reaction is preferably room temperature, and the time is preferably 2 hours.

[0102] In this invention, the catalyst preferably comprises triphenylphosphine gold trifluoromethane sulfonate (PPh3AuOTf). In this invention, the PPh3AuOTf is preferably purchased or prepared in-house. When prepared in-house, the preparation of the catalyst preferably includes the following steps:

[0103] Under argon protection and in the dark, PPh3AuCl and AgOTf are added sequentially to dichloromethane, and the reaction is carried out for 10 minutes. In this invention, the molar ratio of PPh3AuCl to AgOTf is preferably 1:1. There are no special requirements for the amount of dichloromethane used; it can be adjusted according to the actual reaction needs.

[0104] After obtaining the reaction solution containing the pentasaccharide intermediate, the present invention preferably does not perform post-processing, and then mixes the reaction solution containing the pentasaccharide intermediate, arabinose acceptor 16, and fourth activator to carry out the eighth glycosylation reaction to obtain hexasaccharide 27.

[0105] In this invention, the fourth activator is preferably N-iodosuccinimide and trimethylsilyl trifluoromethanesulfonate; the molar ratio of N-iodosuccinimide and trimethylsilyl trifluoromethanesulfonate is preferably (1.5-1.7):1, and in the embodiments of this invention, it can be 1.5:1, 1.65:1 or 1.7:1.

[0106] In this invention, before mixing the reaction solution containing the pentasaccharide intermediate, arabinose acceptor 16, and the fourth activator, it is preferable to further cool the reaction solution containing the pentasaccharide intermediate. In this invention, the cooling is preferably to -78°C.

[0107] In this invention, the molar ratio of the arabinose receptor 16 to the fourth activator is preferably 1:(9.5-10), and in specific embodiments of this invention it can be 1:9.5, 1:9.86 or 1:10.

[0108] In this invention, the temperature of the eighth glycosylation reaction is preferably -78°C, and the time is preferably 3 hours.

[0109] After completing the eighth glycosylation reaction, the present invention preferably uses Et3N to quench the reaction and then filters, concentrates, and finally separates and purifies by column chromatography.

[0110] After obtaining the hexasaccharide 27, the present invention selectively removes the TIPS protecting group from the hexasaccharide 27 to obtain hexasaccharide 10.

[0111] In this invention, the selective removal of the TIPS protecting group from the hexasaccharide 27 preferably includes the following steps: dissolving the hexasaccharide 27 in a methanol-dichloromethane mixed solvent (MeOH / DCM), cooling, and then adding acetyl chloride (AcCl) dropwise to remove the TIPS protecting group.

[0112] In this invention, the cooling is preferably to -20°C; the removal time of the TIPS protective substrate is preferably 48 hours.

[0113] In this invention, the volume ratio of methanol to dichloromethane in the methanol-dichloromethane mixed solvent is preferably 1:1; the molar amount of hexasaccharide 27 and the volume ratio of the methanol-dichloromethane mixed solvent are preferably 0.07 mmol:0.68 mL; and the concentration of acetyl chloride in the methanol-dichloromethane mixed solvent is preferably 1 mol / L.

[0114] After removing the TIPS protecting group, the present invention preferably quenches the reaction with saturated NaHCO3 solution, then extracts three times with DCM, washes the organic phase with water and saturated brine, dries it with anhydrous Na2SO4, filters, concentrates, and finally separates and purifies it by column chromatography.

[0115] In this invention, the arabinose ABz receptor 14 has the structure shown in Formula 14; the arabinose PVB receptor 15 has the structure shown in Formula 15; and the arabinose receptor 16 has the structure shown in Formula 16.

[0116]

[0117] The present invention does not have any special requirements for the preparation of the compounds with structures shown in Formulas 14 to 17; they can be obtained by means well known in the art.

[0118] After obtaining the nine-saccharide 31, the present invention selectively removes the Bz and Lev protecting groups in the nine-saccharide 31 and performs hydrogenolysis on the Bn and Cbz groups to obtain the substructure of the repeating unit of the Angelica polysaccharide APS-1 II.

[0119] In this invention, the selective removal of the Bz and Lev protecting groups in the nonaglycone 31 preferably includes: dissolving the nonaglycone 31 in a methanol-dichloromethane mixed solvent, then adding sodium methoxide (NaOMe), adjusting the pH to 11, stirring at room temperature, then neutralizing the reaction with 4M HCl until the pH is 7, and then filtering, concentrating, separating and purifying to obtain the intermediate.

[0120] In this invention, the volume ratio of methanol to dichloromethane in the methanol-dichloromethane mixed solvent is preferably 1:1; the molar amount of the nonaglycone 31 and the volume ratio of the methanol-dichloromethane mixed solvent are preferably 0.03 mmol: 3 mL.

[0121] After obtaining the intermediate, the present invention performs hydrogenolysis on the Bn and Cbz groups in the intermediate to obtain the substructure 9 sugar.

[0122] In this invention, the hydrogenolysis preferably includes: dissolving the intermediate, palladium on carbon (Pd / C), in a methanol-acetic acid aqueous solution (MeOH / H2O / HOAc), stirring at room temperature under a hydrogen atmosphere, and then filtering, concentrating, separating and purifying.

[0123] In this invention, the mass ratio of the intermediate to palladium on carbon is preferably 1:(1.4-1.6), and in specific embodiments, it can be 1:1.4, 1:1.54, or 1:1.6. In this invention, the volume ratio of methanol, water, and acetic acid is preferably 4 mL:0.5 mL:0.01 mL. In this invention, the molar amount of the intermediate and the total volume ratio of the methanol-acetic acid aqueous solution is preferably 0.024 mmol:4.51 mL. In this invention, the stirring time at room temperature is preferably 10 h.

[0124] In this invention, the method for preparing the 13-saccharide repeating unit of Angelica polysaccharide APS-1 II includes the following steps:

[0125] Nine-sugar 31 was obtained according to the preparation method described above;

[0126] The Lev protecting group of the nonaglycone 31 was removed to obtain nonaglycone 8.

[0127] In this invention, the removal of the Lev protecting group from the nonaglycone 31 preferably includes: dissolving the nonaglycone 31 in a pyridine-acetic acid mixed solvent (pyridine / AcOH), adding hydrazine hydrate, stirring overnight at room temperature, quenching the reaction with acetone, concentrating, then diluting the solution with ethyl acetate, washing with 4M HCl, saturated NaHCO3 solution and saturated brine, drying with anhydrous Na2SO4, filtering, concentrating, and finally purifying by column chromatography.

[0128] In this invention, the volume ratio of pyridine to acetic acid in the pyridine-acetic acid mixed solvent is preferably 3:2; the molar amount of the nonaconitine 31 and the volume ratio of the pyridine-acetic acid mixed solvent are preferably 0.97 mmol:1 mL; the molar ratio of the nonaconitine 31 to hydrazine hydrate is preferably (3-4):1, and in the embodiments of this invention, it can specifically be 3:1, 3.3:1 or 4:1.

[0129] After obtaining the nine-glucose 8, the present invention mixes glucose PTFAI donor 5, glucose ABZ acceptor 6, dehydrating agent, first polar organic solvent and first activator to carry out the ninth glycosylation reaction to obtain a reaction solution containing disaccharide ABZ intermediate.

[0130] In this invention, the first activator is preferably trifluoromethanesulfonic acid; the first polar organic solvent preferably includes dichloromethane; and the dehydrating agent preferably includes a molecular sieve.

[0131] In this invention, mixing glucose PTFAI donor 5, glucose ABZ acceptor 6, dehydrating agent, first polar organic solvent and first activator preferably includes: mixing and stirring glucose PTFAI donor 5, glucose ABZ acceptor 6, dehydrating agent and first polar organic solvent, then cooling, and then adding the first activator dropwise.

[0132] In this invention, the molar ratio of glucose PTFAI donor 5 to glucose ABZ acceptor 6 is preferably 1:1; the molar amount of glucose PTFAI donor 5 and the volume ratio of the first polar organic solvent are preferably 0.04 mmol:0.4 mL; the mass ratio of glucose PTFAI donor 5 to dehydrating agent is preferably 1:(10-12), and in the embodiments of this invention, it can specifically be 1:10, 1:11 or 1:12; the molar ratio of glucose PTFAI donor 5 to the first activator is preferably 4:1.

[0133] In this invention, the mixing and stirring time is preferably 15 min; the cooling is preferably to -40°C; the temperature of the ninth glycosylation reaction is preferably -40°C, and the time is preferably 2 h.

[0134] After obtaining the reaction solution containing the disaccharide ABz intermediate, the present invention preferably does not perform post-processing, and then mixes the reaction solution containing the disaccharide ABz intermediate, arabinose PVB acceptor 7 and catalyst, and performs the tenth glycosylation reaction to obtain the reaction solution containing the tetrasaccharide PVB intermediate.

[0135] In this invention, mixing the reaction solution containing the disaccharide ABz intermediate, the arabinose PVB acceptor 7, and the catalyst preferably includes: heating the reaction solution containing the disaccharide ABz intermediate, and then adding the arabinose PVB acceptor 7 and the catalyst.

[0136] In this invention, the heating is preferably performed to room temperature. In this invention, the arabinose PVB receptor 7 is preferably added in the form of an arabinose PVB receptor 7 solution; the arabinose PVB receptor 7 solution is preferably obtained by dissolving arabinose PVB receptor 7 in dichloromethane; the concentration of the arabinose PVB receptor 7 solution is preferably 0.2 mol / L.

[0137] In this invention, the catalyst is preferably added in the form of a catalyst solution; the catalyst solution is preferably obtained by dissolving the catalyst in dichloromethane; and the concentration of the catalyst solution is preferably 0.05 mol / L.

[0138] In this invention, the catalyst preferably comprises triphenylphosphine gold trifluoromethane sulfonate. In this invention, the PPh3AuOTf is preferably purchased or prepared in-house. When prepared in-house, the catalyst preparation preferably includes: adding PPh3AuCl and AgOTf sequentially to dichloromethane under argon protection and light-protected conditions, and reacting for 10 minutes. In this invention, the molar ratio of PPh3AuCl to AgOTf is preferably 1:1. This invention does not have specific requirements for the amount of dichloromethane used; it can be adjusted according to the actual reaction needs.

[0139] In this invention, the molar ratio of the arabinose PVB receptor 7 (pure substance) to the catalyst (pure substance) is preferably 8:1; the molar ratio of the arabinose PVB receptor 7 (pure substance) to the glucose PTFAI donor 5 is preferably 1:1.

[0140] In this invention, the temperature of the tenth glycosylation reaction is preferably room temperature, and the time is preferably 2 hours.

[0141] After obtaining the reaction solution containing the tetrasaccharide PVB intermediate, the present invention preferably does not perform post-processing, and then mixes the reaction solution containing the tetrasaccharide PVB intermediate, the nonaconitose 8, and the second activator to carry out the eleventh glycosylation reaction to obtain the tridecansaccharide 35.

[0142] In this invention, the second activator is preferably N-iodosuccinimide and trifluoromethanesulfonic acid; the molar ratio of N-iodosuccinimide to trifluoromethanesulfonic acid is preferably 5:1; the molar ratio of the nonaglycone 8 and the second activator is preferably 5:1; the molar ratio of the nonaglycone 8 and arabinose PVB acceptor 7 (pure substance) is preferably 3:0.4. In this invention, the time for the eleventh glycosylation reaction is preferably 2 hours.

[0143] After completing the eleventh glycosylation reaction, the present invention preferably quenches the resulting reaction solution with triethylamine, and then filters, concentrates and purifies it.

[0144] After obtaining the tridecanedose 35, the present invention removes the protecting group from the tridecanedose 35 to obtain the 13-saccharide repeating unit of the Angelica polysaccharide APS-1 II.

[0145] In this invention, the removal of the protecting group from the tridecanose 35 preferably includes: dissolving the tridecanose 35 in a methanol-dichloromethane mixed solvent, then adding sodium methoxide and adjusting the pH to 11, stirring at room temperature for 8 days, then neutralizing the reaction with 4M HCl until the pH is 7, and then filtering, concentrating, separating and purifying to obtain an intermediate; dissolving the intermediate and palladium on carbon in a methanol-acetic acid aqueous solution, stirring at room temperature for 10 hours under a hydrogen atmosphere, and then filtering, concentrating, separating and purifying.

[0146] In this invention, the volume ratio of methanol to dichloromethane in the methanol-dichloromethane mixed solvent is preferably 1:1; the molar amount of tridecanedioic acid 35 and the volume ratio of methanol-dichloromethane mixed solvent are preferably 0.015 mmol: 3 mL.

[0147] In this invention, the mass ratio of the intermediate to palladium on carbon is preferably 1:(2.5-3.5), and in specific embodiments, it can be 1:2.5, 1:2.9, or 1:3.5. In this invention, the volume ratio of methanol, water, and acetic acid is preferably 4 mL:0.5 mL:0.01 mL. In this invention, the molar amount of the intermediate and the total volume ratio of the methanol-acetic acid aqueous solution is preferably 0.014 mmol:4.51 mL.

[0148] After removing the 35 protecting group of the decadecyl sugar, the present invention preferably filters the resulting reaction solution and then concentrates and purifies it.

[0149] In this invention, the nonaglycone 8 has the structure shown in Formula 8; the glucose PTFAI donor 5 has the structure shown in Formula 5; the glucose ABZ receptor 6 has the structure shown in Formula 6; the arabinose PVB receptor 7 has the structure shown in Formula 7; the tridecans 35 has the structure shown in Formula 35; and the 13-saccharide repeating unit 1 has the structure shown in Formula 1.

[0150]

[0151] The present invention does not have any special requirements for the preparation of compounds with the structures shown in Formulas 5 to 8; they can be obtained by means well known in the art.

[0152] This invention provides the application of the Angelica polysaccharide APS-1 II repeating unit and its substructure described in the above-described scheme, or the Angelica polysaccharide APS-1 II repeating unit and its substructure prepared by the preparation method described in the above-described scheme, in the preparation of drugs that inhibit the proliferation of leukemia cells.

[0153] To further illustrate the present invention, the following detailed descriptions, in conjunction with embodiments, provide a highly stereoselective 1,2-cis-fucosylation reaction promoter composition, a fucose PTFAI donor and its application, a method for a highly stereoselective 1,2-cis-fucosylation reaction, the 13-saccharide repeating unit of Angelica polysaccharide APS-1 II, the substructure 9 sugar and its preparation method and application, but these descriptions should not be construed as limiting the scope of protection of the present invention.

[0154] Example 1

[0155] Preparation of 2-O-benzyl-3,4-O-benzoyl-N-trifluoroacetylimine ester-L-pyranose fucose (21d):

[0156]

[0157] S3 (4.46 g, 7.61 mmol) was dissolved in Py (15 mL), and BzCl (2.64 mL, 3 equivalents) and DMAP (185.7 mg, 1.52 mmol, 0.2 equivalents) were added. The reaction was carried out overnight at room temperature. After the reaction was completed, the mixture was quenched with water, extracted with DCM, dried over anhydrous Na2SO4, filtered, and concentrated. The solution was purified by silica gel column chromatography (PE / EA5 / 1) to obtain 27 (4.32 g, 99% yield). 27 (4.32 g, 7.60 mmol) was dissolved in acetone / water (6 1 / 15.2 mL) and cooled to 0 °C. TCCA (1.79 g, 7.60 mmol) was added, and the mixture was allowed to react overnight at room temperature. Extraction was performed using DCM, followed by washing with saturated NaHCO3, water, and saturated drinking water, drying on anhydrous Na2SO4, filtration, and concentration. The above intermediate (400 mg, 0.86 mmol) and 2,2,2-trifluoro-N-phenylacetyl chloride (180 mg, 0.86 mmol) were dissolved in acetone (3.4 mL), and K2CO3 (119 mg, 0.86 mmol) was added. The mixture was allowed to react overnight at room temperature, filtered, and concentrated. Purification was performed by silica gel column chromatography (PE / EA 10 / 1, containing 0.1% Et3N) to give 21d (531 mg, 97% yield). 1 HNMR(400MHz, CDCl3)δ7.94-7.84(m,2H),7.73-7.69(m,2H),7.55-7.49(m,1H),7.41-7.34(m,3H),7.27-7.02(m,10H),6.80-6.71(m,2H),5.70- 5.66(m,1H),5.50(s,1H),5.33(s,1H),4.74(d,J=12.0Hz,1H),4.61(d,J =12..0Hz,1H),4.66-4.61(m,1H),4.10-3.90(m,1H),1.20-1.12(m,3H); 13C NMR (100MHz, CDCl3) δ165.9,165.86,165.6,165.5,143.7,143.5,137.4,137.3,133.5,1 33.49,133.3,133.2,130.0,129.9,129.8,129.7,129.5,129.48,129.4,128.9,128.8,12 8.7,128.5,128.44,128.4,128.38,128.36,128.03,128.0,124.5,119.4,97.5,75.4,75 .0,73.2,73.1,72.5,71.8,71.2,70.6,70.4,68.0,16.3,16.2.HRMS(ESI),m / zcalcd.for C 35 H 30 F3NO7Na[M+Na] + 656.1872, found: 656.1872.

[0158] Example 2

[0159] The synthesis of (trans-3,7-methyl-2,6-octadienyl)-2-O-benzyl-3,4-O-benzoyl-α-L-pyranofucose (25d) is shown in the following reaction equation:

[0160]

[0161] The glycosylation reaction was carried out for 24 h according to the preparation steps of Example 1, except that the donor 21d (88 mg, 0.14 mmol) and the acceptor 24d (32 mg, 0.21 mmol, 1.5 equivalents), Ph3PO (232 mg, 0.83 mmol, 6 equivalents), DCM (1.4 mL) were prepared. Molecular sieve (120 mg) and TMSI (20 μL, 0.14 mmol, 1 equivalent) were used. Purification by silica gel column chromatography (PE / EA = 10 / 1) yielded 25 d (75 mg, 90% yield, α / β > 20:1) as a syrup. [α] D 26 = -170.11 (c 0.27, CHCl3); 1HNMR(400MHz, CDCl3)δ7.96(d,J=7.3Hz,2H),7.82(d,J=7.4Hz,2H),7.59(t,J=7.4Hz,1H),7.47-7.42(m,3H),7.3 1-7.24(m,7H),5.75(dd,J=10.5,3.3Hz,1H),5.66(d,J=2.6Hz,1H),5.44(t,J=6.5Hz,1H),5.12(d,J=6.5Hz,1H), 5.04(d,J=3.4Hz,1H,1-H),4.67(d,J=12.0Hz,1H),4.64(d,J=12.0Hz,1H),4.36(q,J=6.3Hz,1H),4.26-4.16(m,2 H),4.12(dd,J=10.5,3.5Hz,1H),2.29-2.06(m,2H),1.72(s,1H),1.70(s,2H),1.63(s,1H),1.20(d,J=6.5Hz,2H); 13 C NMR (100MHz, CDCl3) δ165.9,165.5,141.2,138.0,133.2,132.8,131.8,130.0,129.8,129.6,128.4,128.3,128.2,128.0 ,127.8,123.9,119.9,96.0(1-C),73.3,72.6,70.7,64.8,64.2,39.7,26.5,25.7,17.7,16.5,16.2.HRMS(ESI)calcdfor C 37 H 42 O7Na[M+Na] + 621.2823, found 621.2820.

[0162] Example 3

[0163] The synthesis of methyl-3-O-(2-O-benzyl-3,4-O-benzoyl-α-L-pyranofucrosyl)-2,3,6-O-benzyl-α-D-pyranose (25m) is shown in the following reaction equation:

[0164]

[0165] The glycosylation reaction was carried out for 52 hours following the preparation steps of Example 1, with the only difference being the donor 21d (158.6 mg, 0.25 mmol, 1.5 equivalents) and acceptor 24m (77.5 mg, 0.17 mmol), Ph3PO (418 mg, 1.5 mmol, 9 equivalents), and DCM (1.7 mL). Molecular sieve (230 mg) and TMSI (34 μL, 0.25 mmol, 1.5 equivalents) were purified by silica gel column chromatography (PE / EA = 5 / 1) to 25 mg (127 mg, yield 96%, α / β > 20:1) as a white solid. [α] D 25.2 = -128.57 (c 0.35, CHCl3); 1 H NMR (400MHz, CDCl3) δ7.77(d,J=7.4Hz,2H),7.70(d,J=7.5Hz,2H),7.46-6.95(m,2 6H),5.77(s,1H,1-H),5.73(d,J=10.6Hz,1H),5.16(s,1H),4.67(q,J=10.9Hz,2H) ,4.60-4.55(m,5H,1'-H),4.44(d,J=11.9Hz,1H),4.38-4.32(m,2H),4.24(t,J=8. 8Hz,1H),4.98-3.95(m,1H),3.78-3.65(m,5H),3.24(s,3H),0.59(d,J=5.9Hz,3H); 13 C NMR (100MHz, CDCl3) δ166.0,165.6,138.3,138.0,137.8,137.7,133.1,132.9, 130.1,129.9,129.8,129.7,128.6,128.5,128.4,128.3,128.2,128.1,127.9,1 27.84,127.8,127.65,127.6,97.7(1'-C),96.6(1-C),81.7,77.0,75.4,74.1,7 3.7,73.0,72.7,72.5,72.3,70.5,70.4,68.8,64.5,55.2,15.6.HRMS(ESI),m / z calcd.For C 55 H 56 O 12 Na[M+Na] + 931.3664, found 931.3666.

[0166] Example 4

[0167] The synthesis of simvastatin-2-O-benzyl-3,4-O-benzoyl-α-L-pyranofucose (25v) is shown in the following reaction equation:

[0168]

[0169] The glycosylation reaction was carried out for 4.5 days following the preparation steps in Example 1, with the only differences being the donor 21d (124 mg, 0.20 mmol) and acceptor 24v (122 mg, 0.29 mmol, 1.5 equivalents), Ph3PO (333.6 mg, 1.20 mmol, 6 equivalents), and DCM (2.0 mL). Molecular sieve (240 mg) and TMSI (28.4 μL, 0.20 mmol, 1 equivalent). Purified by silica gel column chromatography (PE / EA = 3 / 1) to 25 v (137 mg, yield 80%, α / β > 20:1) as a white solid. [α] D 20.1 = -36.50 (c 0.24, CHCl3); 1 HNMR (400MHz, CDCl3) δ7.89(d,J=7.5Hz,2H),7.73(d,J=7.5Hz,2H),7.53(t,J=7.4Hz,1H),7.42-7.36(m,3H),7.23-7.17(m,7H),5.90(d,J =9.6Hz,1H),5.69(dd,J=9.2,6.3Hz,1H),5.61-5.59(m,2H),5.41(s,1H),5.23(d,J=2.6Hz,1H),5.02(d,J=3.4Hz,1H,1-H),4.64-4.57(m, 3H),4.25(q,J=6.2Hz,1H),4.15(s,1H),4.08-4.05(m,1H),2.78(dd,J=17.3,3.2Hz,1H),2.68(dd,J=17.4,5.1Hz,1H),2.35-2.24(m,2H), 2.14-2.06(m,2H),1.84-1.76(m,4H),1.58-1.35(m,6H),1.13(d,J=6.5Hz,3H),1.05(s,6H),1.00(d,J=7.4Hz,3H),0.76(t,J=7.6Hz,6H); 13C NMR (100MHz, CDCl3) δ178.1,169.9,166.2,165.8,138.3,133.7,133.4,133.3,1 31.9,130.2,130.1,130.0,128.9,128.81,128.8,128.6,128.3,128.1,97.3(1-C ),77.8,73.7,73.3,72.6,70.9,69.9,68.2,66.1,43.4,38.0,37.3,37.0,33.6,3 3.4,33.3,31.0,27.7,25.21,25.2,24.1,23.4,16.5,14.3,9.8.HRMS(ESI)calcd for C 52 H 62 O 11 Na[M+Na] + 885.4184, found 885.4182.

[0170] Example 5: Synthesis of substructure 2. Synthesis of acceptor 10: 1) p-methoxyphenyl (2-O-benzoyl-5-O-triisopropylsilyl-α-L-furanarabinose) (20);

[0171]

[0172] The above-mentioned hemiacetal intermediate S8 (S8 reference: Zhang Y, Chen Z, Huang Y, He S, Yang X, Wu Z, Wang X, Xiao G. Modular Synthesis of Nona-Decasaccharide Motif from Psidiumguajava Polysaccharides: Orthogonal One-Pot Glycosylation Strategy. Angew ChemInt Ed Engl. 2020, 59, 7576-7584.) (3.64 g, 6.60 mmol) and ABzOH (1.85 g, 9.91 mmol) were dissolved in dry DCM (33 mL), and DMAP (807 mg, 6.60 mmol) and EDCI (2.28 g, 11.89 mmol) were added. The mixture was stirred at room temperature for 5 hours. After the reaction was complete, the mixture was concentrated. The mixture was purified by column chromatography (petroleum ether / ethyl acetate, 6 / 1) to give the intermediate (4.50 g, 95%, α / β3:1). The above intermediate (1.98 g, 2.76 mmol) and 4-methoxyphenol (377 mg, 3.04 mmol) were added to a dry DCM (26.6 ml). Molecular sieve (2.53 g). After stirring at room temperature for 15 minutes, freshly prepared PPh3AuOTf was added to CH2Cl2 solution (1 mL, 0.44 M). The reaction was stirred overnight, then quenched with Et3N (1 mL), filtered, and concentrated. Column chromatography was used for purification (petroleum ether / ethyl acetate, 10 / 1) to give an intermediate (1.81 g, quant). The above intermediate (1.81 g, 2.76 mmol) was then dissolved in DCM / H2O (27.5 mL, v / v 10 / 1), and DDQ (689 mg, 3.04 mmol) was added. The reaction mixture was stirred at room temperature for 10 hours, diluted with EtOAc, and washed with saturated Na2S2O3 solution, water, and saturated brine. The mixture was dried over anhydrous Na2SO4, filtered, and concentrated. Column chromatography was used for purification (petroleum ether / ethyl acetate, 5 / 1) to give 20 g of colorless syrup (1.54 g, 77%). [α] D 25 = -27.88 (c 0.16, CHCl3). 1 H NMR (400MHz, CDCl3) δ8.05(d,J=7.0Hz,2H),7.60(t,J=7.5Hz,1H),7.46(t,J=7.8Hz,2H),7.05(d,J=9.0Hz,2H),6.84(d,J=9.2Hz,2H),5.79(s, 1H,H-1),5.40(dd,J=3.0,1.4Hz,1H),4.40(m,1H),4.35(m,1H),3.97(d ,J=4.4Hz,2H),3.78(s,3H),3.34(d,J=4.4Hz,1H),1.19-0.95(m,21H). 13 C NMR (100MHz, CDCl3) δ165.96,154.28,149.55,132.81,129.02,128.23,127.66,117.44,113.73,104. 05,85.84,83.88,76.55,76.23,75.91,75.87,62.05,54.80,17.09,17.08,11.10.HRMS(ESI)calcdfor C 68 H 64 O 12 Na[M+Na] + 1095.4290, found 1095.4299.

[0173] 2) o-(1-styryl)-2,3,4-O-benzyl-β-D-glucopyranose benzoate (19)

[0174]

[0175] A solution of S9 (Shen K, Lowary TL. Synthesis of the Mycobacterium tuberculosis Canetti Lipooligosaccharide II Nonasaccharide. Org Lett. 2022, 24, 6428-6432) (4.30 g, 6.25 mmol) and PVBOH (2.10 g, 9.38 mmol) in dry DCM (31.3 ml) was added to DMAP (764 mg, 6.25 mmol), EDCI (1.75 g, 11.25 mmol), and DIPEA (3.30 ml, 18.75 mmol). The resulting mixture was stirred at room temperature for 10 hours. After completion, it was concentrated. Column chromatography was used for purification (petroleum ether / ethyl acetate, 30:1-20:1) to give colorless syrup 19 (4.11 g, 91%, α / β = 1.5 / 1). The intermediate (1.84 g, 2.06 mmol, β isomer) was then dissolved in dry THF (10 mL), and HF / pyridine (70%, 1.8 mL, 20.6 mmol) was added dropwise. The resulting mixture was stirred at room temperature for 4 hours, then quenched with Et3N, diluted with ethyl acetate, washed with saturated NaHCO3 solution and saturated brine, dried over anhydrous Na2SO4, and concentrated. Column chromatography (petroleum ether / ethyl acetate, 5 / 1) was used to purify the mixture, yielding colorless syrup 19 (1.16 g, 86%). [α] D 25 = -18.00 (c 0.18, CHCl3). 1 H NMR (400MHz, CDCl3) δ7.83(d,J=6.4Hz,1H),7.46(td,J=7.5,1.4Hz,1H),7.35-7.25(m,2H),7.24-7.06(m,20H),5.64(s,1H),5.52( d,J=8.0Hz,1H),5.16-5.06(m,1H),4.81-4.66(m,3H),4.66-4.50(m,3H),3.65-3.55(m,2H),3.52-3.36(m,3H),3.32-3.24(m,1H). 13C NMR (100MHz, CDCl3) δ165.2,149.5,143.70,140.74,138.4,138.1,138.0,132.7,132.0,130.7,129.4,128.6,128.6,128.5,128.30,128.2,128 .13,128.07,128.0,127.9,127.9,127.6,126.7,114.3,94.5,84.7,81.1,77.6,77.2,77.0,76.9,75.8,75.7,75.2,75.0,61.4.HRMS(ESI)calcd for C 42 H 40 O7Na[M+Na] + 679.2666, found 679.2672.

[0176] 3) The synthesis of trisaccharide (17) is shown in the following reaction equation:

[0177]

[0178] PTFAI donor 18 (255 mg, 0.42 mmol), PVB receptor 19 (230 mg, 0.35 mmol), and newly activated PTFAI were used in combination. MS (300mg), MS (223 mg) was stirred in dry DCM (3.2 mL) for 15 min and then cooled to -40 °C. TMSOTf (19 μL, 0.11 mmol) was added dropwise to the mixture. After stirring at -40 °C for another 2 h, the reaction mixture was heated to 0 °C and DMF (362 μL, 4.67 mmol) was added using a known procedure. After stirring at 0 °C for 15 min, NIS (99 mg, 0.44 mmol) and TMSOTf (80 μL, 0.44 mmol) were added. After 1.5 h, receptor 20 (151 mg, 0.29 mmol) was dissolved in DCM (1 mL) and added to the reaction system. The reaction mixture was heated to room temperature. The resulting mixture was stirred at room temperature for 10 h, then quenched with Et3N (1 mL) and filtered, concentrated, and purified by column chromatography (petroleum ether / ethyl acetate = 6 / 1) to give colorless syrup 26 (230 mg, 58% yield). 26 (2.52 g, 1.85 mmol) was dissolved in toluene / acetonitrile / H₂O (total volume 37 mL, v / v / v = 3 / 5 / 3), cooled to 0 °C, and then cerium ammonium nitrate (CAN) (2.31 g, 4.21 mmol) was added to remove the anomeric p-methoxyphenyl (MP) group of 26, generating a hemiacetal. After stirring at 0 °C for 2 hours, the reaction mixture was quenched with saturated NaHCO₃ solution, extracted three times with DCM, the organic phase was washed with saturated brine, dried over Na₂SO₄, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate v / v = 4 / 1) was used to purify the mixture into a pale yellow syrup intermediate (1.92 g, 83%). The above intermediate (1.57 g, 2.04 mmol) was dissolved in acetone (15.3 mL), and then 2,2,2-trifluoro-N-phenylacetimidoylchloride (568 mg, 2.75 mmol) and anhydrous potassium carbonate (378 mg, 2.75 mmol) were added for substitution reaction. The mixture was stirred at room temperature for 10 hours. After the reaction was complete, the reaction mixture was filtered and concentrated. Column chromatography was used for purification (petroleum ether / ethyl acetate = 50 / 1, containing 3% Et3N) to give colorless syrup 17 (2.18 g, quant (quantitative yield, 100%), α / β > 20 / 1). 1HNMR(400MHz, CDCl3) δ7.99(d,J=8.3Hz,2H),7.92(d,J=8.3Hz,2H),7.54(q,J=7.7Hz,2H),7.43-7.14 (m,34H),7.07(t,J=7.5Hz,1H),6.88(d,J=7.7Hz,2H),5.74(s,1H),5.38(s,1H),5.11(s,1H),5.06(s, 1H),4.95(t,J=10.0Hz,2H),4.84(d,J=11.0Hz,1H),4.71(q,J=9.6Hz,5H),4.62-4.40(m,5H),4.26(s ,1H),4.07(s,3H),3.99-3.84(m,4H),3.81-3.67(m,2H),3.56(dd,J=14.9,10.6Hz,4H),1.03(s,21H). 13 C NMR (100MHz, CDCl3) δ164.46,164.10,143.01,138.10,137.93,137.61,137.27,137.02,132.58,132.37,129.04,129.00,128.95, 128.71,128.33,127.84,127.67,127.58,127.55,127.49,127.42,127.15,127.09,127.02,126.86,126.82,126.78,126.74,126.7 2,126.69,126.65,126.53,123.30,118.82,105.58,97.31,85.34,82.28,81.21,80.95,80.64,80.54,79.09,76.69,76.55,76.43, 76.23,75.91,74.72,74.02,72.58,72.07,71.30,70.02,68.03,65.03,61.81,28.91,17.13,17.09,11.09,11.01.HRMS(ESI)calcd for C 82 H 90 NO 16 SiNa[M+Na] + 1452.5900, found 1452.5900.

[0179] 4) Preparation of o-(1-styryl)-2-O-benzoyl-5-O-acetylpropionyl-α-L-furanarabinose benzoate (15):

[0180]

[0181] A solution of S8 (1.07 g, 1.95 mmol) and o-(1-phenylenyl)benzoic acid (484 mg, 2.14 mmol) in dry DCM (9.8 ml) was added to DMAP (238 mg, 1.95 mmol), EDCI (433 mg, 3.50 mmol), and DIPEA (1.02 ml, 5.84 mmol). The resulting mixture was stirred at room temperature for 10 hours. After the reaction was complete, the mixture was concentrated. The intermediate was purified by column chromatography (petroleum ether / ethyl acetate, 30:1-10:1) to give 1.42 g, 96%, α / β 3:1. The intermediate (1.00 g, 1.32 mmol, α isomer) was then dissolved in MeOH / DCM (13.2 mL, v / v 1 / 1), cooled to 0 °C, and AcCl (0.94 mL, 13.2 mmol) was added dropwise. After stirring at 0 °C for 5 hours, the reaction mixture was quenched with saturated NaHCO3 solution, extracted three times with DCM, and the combined organic layers were washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated. The intermediate was purified by column chromatography (petroleum ether / ethyl acetate, 4 / 1) to give 692 mg, 87%. The above intermediate (692 mg, 1.15 mmol) and LevOH (200 mg, 1.72 mmol) were added to a dry DCM container (11.6 mL) along with DMAP (142 mg, 1.15 mmol), EDCI (322 mg, 2.07 mmol), and DIPEA (0.60 mL, 3.45 mmol). The resulting mixture was stirred at room temperature for 10 hours. After the reaction was complete, the mixture was concentrated. The intermediate was purified by column chromatography (petroleum ether / ethyl acetate, 3 / 1) to give 803 mg, quant. The intermediate (1.30 g, 1.86 mmol) was then dissolved in DCM / H₂O (18.7 mL, v / v 10 / 1) and DDQ (465 mg, 2.05 mmol) was added. The reaction mixture was stirred at room temperature for 10 hours, diluted with ethyl acetate, and washed with saturated Na₂S₂O₃ solution, water, and saturated brine. The mixture was dried over anhydrous Na₂SO₄, filtered, and concentrated.Column chromatography separation and purification (petroleum ether / ethyl acetate, 1:1) yielded 15g of colorless syrup (888mg, 385%). [α] D 25 =-43.39(c0.13,CHCl3).1HNMR(400MHz,CDCl3)δ8.02(d,J=7.0Hz,2H),7.95(d,J=7.8Hz,1H),7.62 (t,J=7.5Hz,1H),7.58(t,J=7.5Hz,1H),7.48(t,J=7.3Hz,3H),7.37(d,J=7.6Hz,1H),7.33-7.16(m, 5H),6.30(s,1H,H-1),5.81(s,1H),5.26(s,1H),4.87(d,J=2.0Hz,1H),4.29(dd,J=12.1,3.5Hz,1H ),4.20(dd,J=12.0,4.8Hz,1H),4.07(m,2H),2.72(t,J=6.5Hz,2H),2.63-2.54(m,2H),2.15(s,3H). 13 C NMR (100MHz, CDCl3) δ205.8,171.7,165.9,165.4,148.1,142.0,139.2,133.1,131.5,130.6,129.6,129.1,127.8,1 27.6,127.1,127.0,125.8,113.5,99.1,85.1,81.5,76.6,76.3,76.2,75.9,62.1,37.1,29.0,27.0.HRMS(ESI)calcd for C 32 H 30 O9Na[M+Na] + 581.1782, found 581.1787.

[0182] The synthesis of hexasaccharide (27) is shown in the following reaction equation:

[0183]

[0184] The PTFAI donor 17 (126 mg, 0.09 mmol), ABz receptor 14 (14 mg, 0.09 mmol), and newly activated PTFAI were prepared. MS (200mg) (207 mg) was stirred in dry DCM (0.4 mL) at room temperature for 15 minutes, then cooled to -40 °C. TMSOTf (4.0 μL, 0.02 mmol) was added dropwise to the mixture. After stirring at -40 °C for another 2 hours, the reaction mixture was heated to room temperature, and then PVB receptor 15 (41 mg, 0.07 mmol) was dissolved in dry DCM (0.2 mL), and freshly prepared PPh3AuOTf dissolved in DCM (0.1 mL, 0.12 M) was added. After stirring at room temperature for 2 hours, the reaction mixture was cooled to -78°C, and then receptor 16 (47 mg, 0.07 mmol), NIS (98 mg, 0.43 mmol), and TMSOTf (47 μL, 0.26 mmol) were added sequentially. The resulting mixture was stirred at -78°C for 3 hours, then quenched with Et3N (1 mL), filtered, concentrated, and purified by column chromatography (petroleum ether / ethyl acetate = 2:1, v / v) to give colorless syrup 27 (78 mg, 45% yield). [α] D 25.0=-6.00(c 0.15,CHCl3).1H NMR(400MHz,CDCl3)δ7.95-7.80(m,14H),7.45(q,J=7.4Hz,4H),7.38-6.97(m,52H),5.50(s,2H),5.40(dd,J=12.0,5.5Hz,4H),5.22(d,J=14.4Hz,2H),5.14-4.98(m,3H),4.85(d,J=11.4Hz,3H),4.81-4.70(m,3H),4.65-4.52(m,6H),4.51-4.30(m,12H),4.28-4.19(m,3H),4.12(m,1H),4.04(m,1H),4.00-3.91(m,2H),3.85(d,J=9.9Hz,1H),3.81-3.53(m,6H),3.51-3.23(m,4H),3.10(m,2H),2.52-2.39(m,2H),2.34(m,2H),1.91(s,3H),1.51-1.32(m,4H),1.18(s,2H),0.86(d,J=5.5Hz,21H). 13C NMR (100MHz, CDCl3) δ205.3,171.4,165.2,165.2,164.7,164.5,164.4,164.3,138.2 ,137.9,137.4,137.3,137.2,137.0,132.5,132.44,132.35,132.3,132.0,129.1,129 .05,128.99,128.9,128.9,128.8,128.7,128.6,128.5,127.63,127.59,127.55,127.52,127.48,127.43,127.37,127.02,126.98,126.95,126.92,126.87,126.81,126.77 ,126.74,126.68,126.5,126.37,105.36,105.0,104.5,104.2,103.8,97.2,82.2,82.1,81.9,81.5,81.3,81.2,81.0,80.6,80.4,79.8,79.3,79.2,78.9,78.4,76.6,76.4 3,76.35,76.2,75.9,74.6,74.1,72.6,72.2,71.2,69.8,67.9,66.4,66.2,64.7,62.1 ,62.0,61.5,37.0,28.9,28.8,28.1,26.9,22.4,17.05,16.98,11.0.HRMS(ESI)calcd forC 149 H 159 NO 37 SiNa[M+Na] + 2605.0252, found 2605.0258.

[0185] 5) The synthesis of hexasaccharide (10) is shown in the following reaction equation:

[0186]

[0187] 27 (175 mg, 0.07 mmol) was dissolved in MeOH / DCM (total volume 0.68 mL, v / v = 1 / 1), cooled to -20 °C, and AcCl (48 μL, 0.68 mmol) was added dropwise to selectively remove the TIPS groups from 27 using AcCl and MeOH. The reaction mixture was stirred for 48 hours, quenched with saturated NaHCO3 solution, and then extracted three times with DCM. The organic phase was washed with water and saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated. Column chromatography was used for purification (petroleum ether / ethyl acetate v / v = 1 / 1) to give 10 (143 mg, 87% yield) of colorless syrup. [α] D 24.2 = -7.00 (c 0.08, CHCl3). 1 H NMR (400MHz, CDCl3) δ8.01-7.73 (m, 14H), 7.46 (t, J = 6.3Hz, 4H), 7.42-6.9 6(m,52H),5.52(s,2H),5.45-5.39(m,2H),5.37(d,J=2.3Hz,2H),5.24(s, 1H),5.19(d,J=1.9Hz,1H),5.13-5.01(m,3H),4.85(d,J=9.9Hz,2H),4.76 (dt,J=10.2,3.1Hz,3H),4.72(d,J=3.4Hz,1H),4.67-4.60(m,2H),4.59-4 .51(m,5H),4.51-4.41(m,4H),4.41-4.29(m,7H),4.28-4.12(m,5H),4.05 (dt,J=7.1,4.0Hz,1H),3.98-3.89(m,2H),3.83(d,J=10.0Hz,1H),3.64(m ,6H),3.43(m,3H),3.34-3.23(m,1H),3.17-2.99(m,2H),2.47(dd,J=8.4, 6.4Hz,2H),2.36(q,J=6.5Hz,2H),1.93(s,3H),1.43(s,4H),1.18(s,2H). 13C NMR (100MHz, CDCl3) δ206.4,172.5,166.4,166.2,165.6,165.55,165.48,16 5.43,165.3,139.0,138.7,138.3,138.3,138.1,137.9,133.5,133.45,133. 38,133.3,133.2,133.0,130.05,129.97,129.9,129.8,129.6,129.5,129.4,128.7,128.63,128.58,128.5,128.45,128.38,128.2,128.03,127.97,127. 9,127.8,127.7,127.6,127.6,127.3,106.3,106.0,105.3,105.1,105.0,98.3,83.2,82.9,82.5,82.2,81.9,81.6,81.0,80.8,80.7,80.2,80.0,77.6,7 7.4,77.2,76.9,75.7,75.0,73.7,73.5,72.3,71.0,69.0,67.34,67.2,65.7 ,63.0,62.1,51.0,37.9,29.8,29.8,29.1,28.0,27.8,23.4.HRMS(ESI)calcd for C 140 H 139 NO 37 Na[M+Na] + 2448.8918, found2448.8921.

[0188] Synthesis of substructure 2: 6) 2-O-benzyl-3-O-acetylpropionyl-4-O-benzoyl-N-trifluoroacetylimine ester-D-pyranose fucose (12)

[0189]

[0190] Preparation of 24O (1.18 g, 2.54 mmol) and LevOH (443 mg, 3.82 mmol) was dissolved in dry DCM (25 ml), and DMAP (311 mg, 2.54 mmol), EDCI (711 mg, 4.58 mmol), and DIPEA (1.30 ml, 7.63 mmol) were added. The mixture was stirred at room temperature for 10 hours. After the reaction was complete, the mixture was concentrated. The intermediate was purified by column chromatography (petroleum ether / ethyl acetate, 2 / 1) to give 1.38 g, 97%. The intermediate (1.38 g, 2.46 mmol) was dissolved in acetone / H₂O (61 mL, v / v 4 / 1), cooled to 0 °C, and TCCA (571 mg, 2.46 mmol) was added. The mixture was stirred at room temperature for 4 hours. The solution was diluted with ethyl acetate, washed with saturated NaHCO₃, water, and brine, dried over anhydrous Na₂SO₄, filtered, and concentrated. Column chromatography was used for purification (petroleum ether / ethyl acetate, 1 / 1) to give a hemiacetal intermediate (1.03 mg, 92%). The intermediate (1.03 g, 2.26 mmol) was dissolved in acetone (23 mL), and then 2,2,2-trifluoro-N-phenylacetimidoyl chloride (842 mg, 4.07 mmol) and anhydrous potassium carbonate (570 mg, 4.07 mmol) were added. The mixture was stirred at room temperature for 10 hours. After the reaction was complete, the reaction mixture was filtered and concentrated. Column chromatography separation and purification (petroleum ether / ethyl acetate, 60 / 1, containing 3% Et3N) yielded 12 g of colorless syrup (1.42 g, quant, β / α = 1.7 / 1). 1HNMR (400MHz, CDCl3) δ8.13(d,J=7.7Hz,3H),8.06(d,J=7.8Hz,2H),7.70-7.63(m,3H),7.53(m,6H),7. 41-7.27(m,19H),7.15(q,J=7.1Hz,3H),6.89(d,J=7.8Hz,3H),6.81(d,J=7.7Hz,2H),5.63(s,1H),5.53 (d,J=3.4Hz,1H),5.51-5.46(m,2H),5.16(s,2H),4.86(d,J=11.2Hz,2H),4.75(t,J=5.6Hz,4H),4.44(s ,1H),4.18-4.10(m,1H),3.99(t,J=9.3Hz,2H),2.85-2.37(m,13H),2.12(d,J=8.0Hz,8H),1.27(m,8H). 13 CNMR(100MHz, CDCl3)δ205.3,205.1,171.0,170.9,165.12,165.06,142.7,142.5,136.7,132. 7,132.6,129.0,128.9,128.5,128.4,127.90,127.86,127.8,127.6,127.5,127.3,127.10,12 7.08,126.99,123.5,123.4,118.6,118.4,76.6,76.2,75.9,74.5,74.2,72.4,72.2,71.6,70. 6,70.0,69.5,69.4,66.8,37.0,36.9,28.80,28.76,27.1,27.0,15.24,15.19.HRMS(ESI)calcd for C 33 H 32 F3NO8Na[M+Na]+650.1972, found 650.1974.

[0191] 7) The synthesis of disaccharide (29) is shown in the following reaction equation:

[0192]

[0193] PTFAI donor 12 (456 mg, 0.67 mmol) and acceptor 11 (i.e., formula 24o) (225 mg, 0.45 mmol) were dissolved in dry DCM (1.12 g, 4.03 mmol) with Ph3P=O (1.12 g, 4.03 mmol) at room temperature under argon protection. Molecular sieve (701 mg) was added and stirred for 15 minutes. Then, TMSI (95 μL, 0.67 mmol) was added dropwise. The reaction was stirred at room temperature for 48 h. After the reaction was complete, the solution was diluted with ethyl acetate, and the reaction was quenched with saturated Na₂S₂O₃ (the mixture was directly separated after quenching). The organic phase was washed successively with water and saturated brine, dried over anhydrous Na₂SO₄, filtered, and concentrated. Column chromatography was used for purification (petroleum ether / ethyl acetate = 4 / 1 v / v) to obtain colorless syrup 28 (429 mg, 98% yield). 28 (429 mg, 0.48 mmol) was dissolved in pyridine / AcOH (total volume 4.7 mL, v / v = 3 / 2) and NH₂NH₂·H₂O (69 μL, 1.43 mmol) was slowly added. After stirring overnight at room temperature, the reaction was quenched with acetone, concentrated, and then the mixture was diluted with ethyl acetate and washed successively with 4 M HCl, saturated NaHCO₃ solution, and saturated brine. The mixture was dried over anhydrous Na₂SO₄, filtered, and concentrated. Column chromatography separation and purification (petroleum ether / ethyl acetate, 3 / 1) yielded 29 (377 mg, 99% yield). [α] D 25.1 = -109.50 (c 0.24, CHCl3). 1 HNMR (400MHz, CDCl3) δ8.02(d,J=7.0Hz,2H),7.90(d,J=7.0Hz,2H),7.64(d,J=8.1Hz,2H),7.57(dt,J=21.3,6.8Hz,2H),7.46-7.29( m,9H),7.24-7.16(m,5H),7.12(dd,J=6.7,2.7Hz,2H),5.71(d,J=3.2Hz,1H),5.43(d,J=3.4Hz,1H,1-H),5.07(d,J=10.4Hz,1H),5.03 (d,J=3.3Hz,1H),4.68(d,J=9.5Hz,1H,2-H),4.58(d,J=10.4Hz,1H),4.53(d,J=12.0Hz,1H),4.35(d,J=12.0Hz,1H),4.18(q,J=6.5H z,1H),4.15-4.04(m,2H),3.92-3.81(m,2H),3.75(dd,J=10.1,3.3Hz,1H),2.43(s,3H),1.32(d,J=6.3Hz,3H),0.91(d,J=6.4Hz,3H). 13C NMR (100MHz, CDCl3) δ166.50,166.47,138.2,138.1,137.64,133.62,133.5,13 3.1,130.3,129.97,129.91,129.89,129.4,129.1,128.7,128.6,128.5,128.4, 128.25,128.17,127.94,127.88,92.0,87.3,77.6,77.2,76.9,75.79,75.75,75 .4,75.2,73.6,73.5,72.0,68.7,67.9,65.2,21.4,17.1,16.0.HRMS(ESI)calcd for C 47 H 48 O 10 SNa[M+Na] + 827.2860, found 827.2869.

[0194] 8) The synthesis of trisaccharides (9) is shown in the following reaction equation:

[0195]

[0196] PTFAI donor 21d (445 mg, 0.70 mmol) and acceptor 29 (377 mg, 0.47 mmol) were dissolved in dry DCM (1.17 g, 4.22 mmol) with Ph3P=O (1.17 g, 4.22 mmol) at room temperature under argon protection. Newly activated PTFAI was then added. Molecular sieve (879 mg) was added and stirred for 15 minutes. Then, TMSI (67 μL, 0.47 mmol) was added dropwise. The reaction was stirred at room temperature for 96 h. After the reaction was complete, the solution was diluted with ethyl acetate, and the reaction was quenched with saturated Na₂S₂O₃ solution (the mixture was directly separated after quenching). The organic phase was washed successively with water and saturated brine, dried over anhydrous Na₂SO₄, filtered, and concentrated. Column chromatography was used for purification (petroleum ether / ethyl acetate = 3 / 1) to give white foam 30 (546 mg, 94% yield). 30 (233 mg, 0.19 mmol) was dissolved in acetone / H₂O (total volume 4.6 mL, v / v 4 / 1), cooled to 0 °C, and TCCA (43 mg, 0.19 mmol) was added. The mixture was stirred at room temperature for 10 h. The solution was diluted with ethyl acetate, washed successively with saturated NaHCO₃ solution, water, and saturated brine, dried over anhydrous Na₂SO₄, filtered, and concentrated. The intermediate was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 1 v / v) to give a hemiacetal intermediate (193 mg, 90%). The intermediate (84 mg, 0.07 mmol) was dissolved in acetone (0.7 mL), followed by the addition of 2,2,2-trifluoro-N-phenylacetimidoyl chloride (27 mg, 0.13 mmol) and anhydrous potassium carbonate (18 mg, 0.13 mmol). The mixture was stirred at room temperature for 10 hours. After the reaction was complete, the mixture was concentrated. The purified intermediate was then purified by column chromatography (petroleum ether / ethyl acetate = 60 / 1 v / v, containing 3% Et3N) to give colorless syrup 9 (96 mg, quant (quantitative yield, 100%), β / α = 1.7 / 1). 1HNMR(400MHz,CDCl3)δ8.17(d,J=7.7Hz,4H),8.07(d,J=7.7Hz,2H),8.02-7.97(m,2H),7.94(d,J=7.7Hz,4H),7.87(t,J=7.5Hz,5H),7.72-7.55(m,12H),7.55-7.30(m,40H),7.24(m,18H),7.19-7.12(m,6H),7.12-7.02(m,11H),6.93(d,J=7.9Hz,3H),6.87(d,J=7.7Hz,2H),5.88-5.78(m,2H),5.72(s,2H),5.65(m,3H),5.51(m,2H),5.43(d,J=3.5Hz,2H),5.39(d,J=3.2Hz,2H),5.36(d,J=3.5Hz,2H),5.34(d,J=3.5Hz,1H),5.19(dd,J=7.6,3.5Hz,3H),5.03(d,J=10.4Hz,2H),4.86(m,4H),4.72(t,J=10.3Hz,3H),4.61-4.51(m,4H),4.47-4.31(m,12H),4.29(s,1H),4.24(t,J=6.7Hz,3H),4.17-3.99(m,9H),1.30(m,9H),1.10(d,J=6.4Hz,3H),0.97(d,J=6.4Hz,5H),0.85(d,J=6.4Hz,5H),0.76(d,J=6.4Hz,3H). 13C NMR (100MHz, CDCl3) δ166.70,166.68,166.5,166.3,166.2,166.1,165.41,165.3 7,143.7,138.0,137.9,137.8,137.74,133.66,133.59,133.23,133.19,132.9,13 0.3, 130.2, 130.1, 130.04, 129.95, 129.88, 129.86, 129.83, 129.8, 129.7, 129.5, 129.0, 128.9, 128.82, 128.78, 128.76, 128.6, 128.54, 128.50, 128.48, 128.44, 12 8.39,128.22,128.19,128.17,128.12,128.06,127.8,127.7,127.4,124.5,119.5,93.6,93.1,92.7,92.5,77.6,77.4,77.2,77.1,76.9,76.1,74.3,74.1,73.7,73. 5,73.2,72.7,72.3,72.2,71.85,71.82,70.9,70.5,70.4,70.3,70.2,69.6,69.5, 68.7,68.6,65.3,64.7,64.6,29.9,16.6,16.3,16.1,16.0,15.9.HRMS(ESI)calcd for C 75 H 70 F3NO 17 Na[M+Na] + 1336.4488, found 1336.4495.

[0197] 9) The synthesis of nonaconitose (31) is shown in the following reaction equation:

[0198]

[0199] PTFAI donor 9 (206 mg, 0.16 mmol) and acceptor 10 (254 mg, 0.11 mmol) were dissolved in Ph3P=O (263 mg, 0.95 mmol) in dry DCM (1.6 mL), and freshly activated PTFAI was added under argon protection at room temperature. MS (500 mg), stirred for 15 minutes. Then, TMSI (22 μL, 0.16 mmol) was added dropwise. The reaction was stirred at room temperature for 38 h. After the reaction was complete, the solution was diluted with ethyl acetate, and the reaction was quenched with saturated Na₂S₂O₃ (the phase was separated directly after quenching). The organic phase was washed successively with water and saturated brine, dried over anhydrous Na₂SO₄, filtered, and concentrated. Column chromatography was used for purification (petroleum ether / ethyl acetate v / v = 1.2 / 1) to give white foam 31 (353 mg, 95% yield). [α] D 25.0 = -87.34 (c 0.09, CHCl3). 1 HNMR(400MHz, CDCl3)δ8.10-7.80(m,22H),7.71-7.63(m,2H),7.61-7.50(m,6H),7.50-7.11(m,70 H),7.10-7.00(m,5H),5.62(dt,J=12.4,3.7Hz,5H),5.56(d,J=3.1Hz,1H),5.50(s,1H),5.48(d,J =1.8Hz,1H),5.38(d,J=3.3Hz,1H),5.35(s,1H),5.33(d,J=3.6Hz,1H,1-H),5.29(s,1H),5.27(d, J=3.6Hz,1H,2-H),5.16(q,J=7.2Hz,3H),5.07(d,J=3.5Hz,1H,3-H),4.97(dd,J=7.2,3.7Hz,1H), 4.94(s,1H),4.92-4.81(m,3H),4.78-4.57(m,12H),4.57-4.50(m,3H),4.50-4.27(m,20H),4.06( q,J=3.8Hz,3H),4.04-3.95(m,5H),3.82(d,J=10.6Hz,3H),3.78-3.70(m,1H),3.58(dd,J=9.7,3. 5Hz,1H),3.48-3.35(m,3H),3.21(m,2H),2.62-2.50(m,2H),2.44(q,J=7.0Hz,2H),2.01(s,3H),1 .64-1.45(m,4H),1.30(m,2H),1.07(d,J=6.4Hz,3H),0.92(d,J=6.4Hz,3H),0.69(d,J=6.4Hz,3H). 13C NMR (100MHz, CDCl3) δ206.2,172.3,166.5,166.2,166.13,166.09,166.0,165.6,165.4,165.31,165.28,165. 2,139.0,138.7,138.5,138.2,138.1,138.0,137.92,137.88,136.9,133.3,133.2,133.1,133.0,132.9,132.7 ,130.03,129.97,129.94,129.90,129.87,129.85,129.80,129.7,129.6,129.6,129.5,129.4,128.5,128.5,128.42,128.39,128.34,128.32,128.29,128.25,128.23,128.1,128.03,128.00,127.95,127.90,127.86,127. 83,127.75,127.63,127.61,127.5,127.43,127.39,127.3,106.3,105.9,105.4,105.0,98.8,98.1,93.2,92.4,82.82,82.76,82.6,82.1,81.8,81.5,80.8,80.6,80.3,80.1,79.9,77.4,77.3,77.1,76.8,75.5,75.0,74.2 ,73.9,73.4,72.9,72.83,72.80,72.7,72.2,72.1,71.6,71.0,70.8,70.3,70.2,69.6,68.6,67.2,67.1,65.9, 65.8,65.3,65.0,64.4,63.0,62.8,37.8,29.7,29.7,28.9,27.7,23.3,16.4,16.2,15.7.MS(MALDI-TOF)calcd forC 207 H 203 NO 53 Na[M+Na] + 3473.3113, found 3473.3112.

[0200] 10) The synthesis of nonaconitose (2) is shown in the following reaction equation:

[0201]

[0202] 31 (104 mg, 0.030 mmol) was dissolved in MeOH / DCM (total volume 3 mL, v / v = 1:1), and NaOMe was added to adjust the pH to 11. After stirring at room temperature for 7 days, the reaction was neutralized with 4M HCl to pH = 7, then filtered and concentrated. The intermediate (83 mg, 80%) was purified using Sephadex™ LH2O (V / V ratio MeOH / DCM = 1:1). The intermediate (83 mg, 0.024 mmol) and Pd / C (128 mg, 10%) were dissolved in MeOH / H2O / HOAc (4 mL / 0.5 mL / 0.01 mL), stirred at room temperature under H2 atmosphere for 10 h, then filtered and concentrated. The mixture was purified using Sephadex™ LH-20 (V / V ratio MeOH / H2O = 1 / 1) to give white solid 2 (27.3 mg, yield 82%). [α] D 25.1 = -2.92 (c 0.41, H2O). 1 H NMR(600MHz,D2O)δ5.12(s,1H),5.10(s,1H),5.09(s,1H),5.02(d,J=3.8Hz,1H),4.98(d,J=4.0Hz,1H),4.96(d d,J=4.2,1.7Hz,2H),4.93-4.89(m,2H),4.27(dd,J=11.3,3.8Hz,3H),4.23(s,1H),4.22-4.19(m,1H),4.18(dd ,J=2.7,1.3Hz,1H),4.14(t,J=2.2Hz,1H),4.08-3.93(m,12H),3.93-3.81(m,8H),3.81-3.61(m,12H),3.60-3. 46(m,4H),3.42(t,J=9.5Hz,1H),2.92(t,J=7.6Hz,2H),1.60(m,4H),1.38(q,J=8.1Hz,2H),1.20-1.11(m,9H). 13C NMR(150MHz,D2O)δ107.9,107.2,107.0,106.9,99.3,98.6,95.6,95.2,84.5,83.5,82.62,8 2.56,82.41,81.40,81.39,81.0,79.9,79.5,79.2,79.18,76.24,74.8,74.4,72.8,71.8,71 .13,71.10,69.4,69.3,68.4,68.2,68.0,67.8,67.7,66.9,66.8,66.6,66.54,66.50,66.4, 66.2,60.9,60.88,60.75,39.3,28.0,26.4,23.2,22.2,15.43,15.38,15.2.HRMS(ESI)calcd forC 54 H 93 NO 38 H[M+H] + 1386.5268, found 1386.5264.

[0203] Example 6: Synthesis of 13 sugars 1) Disaccharides (7)

[0204]

[0205] S10 (1.99 g, 2.13 mmol) was dissolved in Toluene / MeCN / H2O (42 ml, v / v / v 3 / 5 / 3) and cooled to 0 °C. Then, CAN (2.67 g, 4.86 mmol) was added. After stirring at 0 °C for 2 hours, the solution was quenched with saturated NaHCO3 solution, extracted three times with DCM, washed with saturated brine, dried over Na2SO4, filtered, and concentrated. The intermediate was purified by column chromatography (petroleum ether / ethyl acetate, 5 / 1) to give a pale yellow syrup intermediate (1.64 g). The intermediate (1.64 g) and PVBOH (666 mg, 2.98 mmol) were dissolved in dry DCM (9.9 ml), and DMAP (242 mg, 1.98 mmol), EDCI (554 mg, 3.57 mmol), and DIPEA (1.04 ml, 5.95 mmol) were added. The resulting mixture was stirred at room temperature for 10 h. After stirring, the solvent was evaporated under vacuum. The intermediate was purified by column chromatography (petroleum ether / ethyl acetate, 15 / 1) to give an intermediate (1.84 g, 83%, 2-step, α / β 1.5:1). The intermediate (324 mg, 0.31 mmol, α isomer) was then dissolved in MeOH / DCM (3.2 ml, v / v 3 / 1), cooled to 0 °C, and AcCl (0.22 ml, 3.14 mmol) was added dropwise. After stirring at 0℃ for 9 h, the reaction was quenched with saturated NaHCO3 solution, extracted three times with CH2Cl2, the organic phase was washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated. Column chromatography (petroleum ether / ethyl acetate, 2 / 1) was used to purify the solution, yielding 7 (233 mg, 85%). [α] D 25 = -91.69 (c 0.52, CHCl3). 1HNMR(400MHz, CDCl3)δ8.14-8.04(m,4H),8.03-7.98(m,1H),7.65-7.56(m,3H),7.51-7.22(m,22H),6.34(s,1H), 5.83(s,1H),5.52-5.47(m,1H),5.42(d,J=1.4Hz,1H),5.32(s,1H),5.29(s,1H),4.86(d,J=11.9Hz,1H),4.68(d,J =12.0Hz,2H),4.52(d,J=12.1Hz,1H),4.39(q,J=4.5Hz,1H),4.28(d,J=4.9Hz,1H),4.20-4.14(m,1H),4.05(d,J= 5.5Hz,1H),3.94(dd,J=11.4,4.2Hz,1H),3.84(dt,J=12.2,3.5Hz,1H),3.75(dd,J=11.4,4.5Hz,1H),3.64(m,1H). 13 C NMR (100MHz, CDCl3) δ166.0,165.3,165.2,149.0,143.2,140.4,137.8,137.6,133.54,1 33.50,132.0,131.3,130.5,130.3,129.9,129.8,129.4,129.2,128.62,128.56,128.4,1 28.2,127.82,127.80,127.77,127.75,127.66,127.63,126.9,114.5,106.4,100.5,84.1 ,83.5,83.0,82.9,81.8,81.0,77.5,77.2,76.9,72.3,72.2,65.7,61.9.HRMS(ESI)calcd for C 53 H 48 O 12 Na[M+Na] + 899.3038, found 899.3039.

[0206] 2) The synthesis of nonaconitose (8) is shown in the following reaction equation:

[0207]

[0208] Dissolve 31 (344 mg, 0.97 mmol) in pyridine / AcOH (total volume 1.0 mL, v / v 3 / 2), and slowly add NH₂NH₂·H₂O (14 μL, 0.29 mmol) to selectively deprotect the Lev group in 31 with NH₂NH₂-H₂O. Stir overnight at room temperature, quench with acetone, concentrate, dilute with ethyl acetate, and wash successively with 4 M HCl, saturated NaHCO₃ solution, and saturated brine. Dry in anhydrous Na₂SO₄, filter, and concentrate. Purify by column chromatography (petroleum ether / ethyl acetate, 1 / 1) to give 8 (333 mg, 97% yield). [α] D 25.3 = -34.43 (c 0.14, CHCl3). 1 H NMR (400MHz, CDCl3) δ8.13-7.77(m,22H),7.66(d,J=7.8Hz,2H),7.60-7.02(m,81H),5 .66-5.54(m,6H),5.52-5.44(m,2H),5.38(d,J=3.3Hz,1H),5.36-5.31(m,2H),5.29(d, J=1.7Hz,1H),5.26(d,J=3.6Hz,1H),5.22-5.11(m,4H),5.07(d,J=3.5Hz,1H),4.96(d ,J=9.5Hz,2H),4.93(d,J=3.4Hz,1H),4.89(d,J=11.2Hz,1H),4.84(d,J=10.8Hz,1H),4 .79(dd,J=12.6,2.6Hz,1H),4.70(q,J=6.3Hz,7H),4.66-4.58(m,4H),4.53(dd,J=6.5 ,3.4Hz,3H),4.49-4.25(m,16H),4.12(q,J=4.4Hz,1H),4.08-3.95(m,7H),3.91-3.68( m,8H),3.57(dd,J=9.6,3.6Hz,1H),3.45(dq,J=11.2,5.8Hz,3H),3.21(m,2H),1.54(s ,4H),1.29(s,2H),1.07(d,J=6.5Hz,3H),0.91(d,J=6.4Hz,3H),0.68(d,J=6.4Hz,3H). 13C NMR (101MHz, CDCl3) δ166.6,166.3,166.3,166.1,165.7,165.54,165.51,165.46,165.4,165.3,13 9.1,138.7,138.5,138.3,138.1,138.1,138.05,138.00,133.4,133.3,133.2,133.0,132.8,130.2 ,130.11,130.08,130.04,130.01,129.98,129.95,129.91,129.88,129.8,129.7,129.6,129.5,128.7,128.7,128.6,128.6,128.53,128.47,128.44,128.41,128.21,128.17,128.15,128.1,128.0, 127.85,127.79,127.76,127.7,127.4,106.5,106.0,105.4,105.0,104.8,98.6,98.2,93.3,92.5,83.2,83.0,82.9,82.6,82.5,82.3,82.2,81.9,81.7,81.3,80.8,80.2,77.6,77.4,77.2,76.9,75. 6,75.23,74.21,74.0,73.5,73.1,73.0,72.7,72.2,71.8,71.0,70.5,70.3,69.8,68.8,67.4,67.3 ,65.5,65.2,64.5,63.4,63.1,61.8,29.9,29.1,28.0,23.4,16.5,16.3,15.8.MS(MALDI-TOF)calcd for C 202 H 197 NO 51 Na[M+Na] + 3475.2745, found 3475.2743.

[0209] 3) The synthesis of tridecanedioic acid (35) is shown in the following reaction equation:

[0210]

[0211] The PTFAI donor 5 (29 mg, 0.04 mmol), ABz receptor 6 (24 mg, 0.04 mmol), and activated... (Reference: B. Yu and H. Tao, Glycosyl Trifluoroacetimidates. 2. Synthesis of Dioscin and Xiebai Saponin IJOrg. Chem. 2002, 67, 9099-9102.) were prepared. MS (151mg) and MS (173 mg) was stirred at room temperature for 15 minutes in dry DCM (0.4 mL), then cooled to -40 °C. TfOH (1.0 μL, 0.01 mmol) was added dropwise to the mixture. After stirring at -40 °C for another 2 hours, the reaction mixture was heated to room temperature, and then PVB receptor 7 (33 mg, 0.04 mmol) dissolved in dry DCM (0.2 mL) and freshly prepared PPh3AuOTf (0.1 mL, 0.05 M in CH2Cl2) were added sequentially. After stirring at room temperature for 2 hours, the reaction mixture was cooled to 0 °C, and then receptor 8 (104 mg, 0.3 mmol), NIS (10 mg, 0.05 mmol), and TfOH (1.0 μL, 0.01 mmol) were added sequentially. The resulting mixture was stirred for 2 hours, then quenched with Et3N (1 mL), filtered, and concentrated. Purification (petroleum ether / ethyl acetate = 2 / 1, v / v) yielded 35g (118mg, 74% yield) of colorless syrup. [α] D 25.3 = -72.75 (c 0.08, CHCl3). 1HNMR(400MHz,CDCl3)δ7.94(m,36H),7.69(d,J=7.8Hz,2H),7.64-6.93(m,122H),5.75(t,J=9.6Hz,1H),5.68-5.52(m,9H),5.49(s,1H),5.37(dd,J=11.7,3.6Hz,4H),5.30(d,J=5.2Hz,2H),5.25(d,J=7.5Hz,2H),5.19(dd,J=10.3,6.7Hz,4H),5.11(d,J=3.6Hz,1H),5.02(d,J=3.5Hz,1H),4.91(m,8H),4.79-4.58(m,14H),4.56-4.16(m,28H),4.13-3.68(m,24H),3.63-3.56(m,2H),3.52(t,J=8.4Hz,2H),3.42(d,J=10.6Hz,4H),3.21(m,3H),1.65-1.44(m,4H),1.31(s,2H),1.08(d,J=6.3Hz,3H),0.93(d,J=6.3Hz,3H),0.73(d,J=6.4Hz,3H). 13C NMR (100MHz, CDCl3) δ166.6,166.4,166.23,166.18,166.1,165.9,165.7,165.6,165.41,165.39,165.34,165.29,165.2,165. 1,165.0,139.11,138.9,138.7,138.4,138.2,138.1,138.1,138.05,138.03,137.97,137.0,133.6,133.4,133.3,133.2,133. 0,132.8,130.2,130.1,130.02,129.98,129.92,129.86,129.83,129.74,129.69,129.66,129.63,129.58,129.1,129.0,128.9,128.7,128.6,128.6,128.53,128.48,128.4,128.32,128.26,128.21,128.15,128.13,128.06,128.02,127.97,127.94,127. 87,127.8,127.74,127.68,127.62,127.58,127.5,127.44,127.39,127.35,106.38,106.0,105.1,105.0,101.4,100.6,99.0,98.2,93.3,92.5,83.4,83.0,82.9,82.6,82.2,82.0,81.8,81.6,81.3,80.7,80.5,80.2,79.62,77.55,77.4,77.2,76.9,75.6, 75.0,74.9,74.6,74.4,74.1,73.1,73.5,73.2,72.9,72.8,72.7,72.4,72.3,72.2,71.9,71.7,71.0,70.7,70.4,70.3,69.9,6 9.8,68.7,67.9,67.6,67.3,65.4,65.1,64.7,64.5,63.0,32.1,29.9,29.0,28.0,23.4,16.5,16.3,15.8.MS(MALDI-TOF)calcd forC 301 H 285 NO 76 Na[M+Na] + 5151.8359, found 5151.8358.

[0212] 4) The synthesis of tridecanedioic acid (1) is shown in the following reaction equation:

[0213]

[0214] 35 (75 mg, 0.015 mmol) was dissolved in MeOH / DCM (total volume 3 mL, v / v = 1:1), and NaOMe was added to adjust the pH to 11. After stirring at room temperature for 8 days, the reaction was neutralized with 4M HCl to pH = 7, then filtered and concentrated. The intermediate (43 mg, 95%) was purified by Sephadex™ LH2O (V / V ratio MeOH / DCM = 1:1) to give the intermediate. The above intermediate (43 mg, 0.014 mmol) and Pd / C (128 mg, 10%) were dissolved in MeOH / H2O / HOAc (4 mL / 0.5 mL / 0.01 mL), stirred at room temperature under H2 atmosphere for 10 hours, then the reaction mixture was filtered and concentrated. 1 (22.5 mg, yield 87%) was purified by Sephadex™ LH-20 (V / V ratio MeOH / H2O = 1 / 1) to give the intermediate. [α] D 24.0 = -587.00 (c 0.12, H2O). 1 H NMR(600MHz,D2O)δ5.13(s,1H),5.11(s,1H),5.07(s,1H),5.04-5.00(m,3H),4.98(d,J=4.1Hz,1H), 4.96(m,2H),4.91(m,2H),4.46(d,J=8.0Hz,1H),4.42(d,J=8.1Hz,1H),4.29-4.11(m,11H),4.07-3. 91(m,14H),3.90-3.80(m,10H),3.80-3.60(m,12H),3.59-3.48(m,5H),3.48-3.37(m,3H),3.34(d,J =9.2Hz,1H),3.31-3.19(m,3H),2.92(t,J=7.6Hz,2H),1.60(m,4H),1.37(m,2H),1.21-1.09(m,9H). 13C NMR(150MHz,D2O)δ107.9,107.5,107.4,107.2,107.2,106.93,106.88,102.5,102.3,99.3,98.6,95.6,95.2,84.3,83.5,82 .6,82.5,82.4,82.2,82.0,81.5,81.39,81.35,81.2,81.0,80.8,80.7,78.0,79.4,79.25,79.16,78.6,76.7,76.6,76.2,75. 9,75.4,74.9,74.7,74.4,74.1,73.1,72.82,72.79,71.9,71.13,71.08,69.4,69.3,68.9,68.5,68.2,68.0,67.8,66.9,66.8 ,66.7,66.5,66.5,66.4,66.3,60.99,60.95,60.7,60.5,59.9,39.3,28.0,26.4,22.2,15.5,15.4,15.2.HRMS(ESI)calcdfor C 76 H 129 NO 56 H[M+H] + 1952.7350, found 1952.7340.

[0215] Activity test

[0216] The effects of compounds 1-2 (i.e., compounds with structures shown in formulas 1-2 of this invention) on the proliferation of different leukemia cell lines were determined using the Cell Counting Kit-8 (CCK-8) method. The method was as follows: Cell culture and administration: Human chronic myeloid leukemia cells K562, mouse leukemia cells L1210, and human monocytes THP-1 were cultured in freshly prepared RPMI 1640 medium (containing 10% FBS and 1% penicillin-streptomycin) at 37°C with 5% CO2. Cells in good condition were collected, centrifuged, and resuspended in fresh medium after discarding the supernatant. The cells were then cultured at 3 × 10⁻⁶ cells / year. 4 Cells were seeded at a density of 100 μL / mL in 96-well plates; blank wells contained no cells but cell culture medium. After 12 h of culture, the drugs were added. The control group received 100 μL / well of culture medium; the compound 1-2 treatment groups received 100 μL / well of drug-containing culture medium, resulting in final drug concentrations of 0.003 mg / L, 0.01 mg / L, 0.03 mg / L, 0.1 mg / L, 0.3 mg / L, 1 mg / L, 3 mg / L, 10 mg / L, and 30 mg / L, respectively. Each group was divided into 6 replicates, and the cells were cultured for another 48 h.

[0217] Absorbance measurement: After 48 hours of drug treatment, 10 μL of CCK-8 was added to each well, and the absorbance at 450 nm was measured using a microplate reader 2 hours later. Data processing: The effect of different concentrations of drug-containing culture medium on the proliferation rate of leukemia cells was calculated. Proliferation rate (%) = (OD value of drug-treated wells - OD value of blank wells) / (OD value of control group - OD value of blank wells) × 100. Statistical analysis: Experimental data are expressed as mean ± standard deviation. Analysis of variance was used to compare different groups. P < 0.05 indicated a statistically significant difference. Specific data are shown in Tables 1 and 2.

[0218] Table 1. Effects of compounds 1-4 on K562 cell proliferation (mean ± standard deviation, n = 6)

[0219]

[0220] Note: *There was a significant difference compared with the control group (P<0.05).

[0221] As shown in Table 1, the inhibitory effects of compounds 1 and 2 on the proliferation of K562 cells were significantly enhanced with increasing drug concentration. When the drug concentration was 30 mg / L, the proliferation rates of K562 cells treated with compounds 1 and 2 were 57.6 ± 1.5% and 50.5 ± 1.2%, respectively (P < 0.05).

[0222] Table 2. Effects of compounds 1 and 2 on the proliferation of L1210 and THP-1 cells (mean ± standard deviation, n = 6)

[0223]

[0224] Note: *There was a significant difference compared with the control group (P<0.05).

[0225] Table 2 shows that the proliferation rates of L1210 cells and THP-1 cells decreased significantly with increasing concentration of compound 2. At a concentration of 30 mg / L, the cell proliferation rates were 53.6 ± 1.4% and 70.0 ± 0.4%, respectively (P < 0.05). Compound 1 also had a significant inhibitory effect on the activity of L1210 cells. At a concentration of 30 mg / L, the cell proliferation rate was 61.2 ± 1.6% (P < 0.05).

[0226] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for preparing a substructure 9 sugar of Angelica polysaccharide APS-1 II, wherein the substructure 9 sugar has the structure shown in Formula 2; Formula 2; The preparation method includes the following steps: Fucose PTFAI donor 12, fucose acceptor 11, first promoter composition, first polar organic solvent and first dehydrating agent are mixed and subjected to a first glycosylation reaction to obtain the first glycosylation product disaccharide 28. The first glycosylation product, disaccharide 28, was deprotected by the Lev protecting group to obtain disaccharide acceptor 29; Fucose PTFAI donor 21d, disaccharide acceptor 29, second promoter composition, second polar organic solvent, and second dehydrating agent are mixed and subjected to a second glycosylation reaction to obtain trisaccharide 30. The protecting group STol was selectively removed from the trisaccharide 30 to obtain a trisaccharide intermediate; the trisaccharide intermediate, 2,2,2-trifluoro-N-phenyliminoacetyl chloride, a third polar organic solvent and a first carbonate were mixed and subjected to a first substitution reaction to obtain a trisaccharide PTFAI donor 9 with a PTFAI protecting group; The trisaccharide PTFAI donor 9, the hexasaccharide acceptor 10, the third promoter composition, the fourth polar organic solvent and the third dehydrating agent are mixed and subjected to a third glycosylation reaction to obtain the nonasugar 31. The Bz and Lev protecting groups in the nine-saccharide 31 are selectively removed, and the Bn and Cbz groups are hydrogenolyzed to obtain the nine-saccharide; the first accelerator composition, the second accelerator composition, and the third accelerator composition comprise triiodosilane and triphenylphosphine oxide; the molar ratio of triiodosilane to triphenylphosphine oxide is 1:(5~10); the stereoselectivity of the glycosylation product is α / β>20:1; The fucose PTFAI donor 12 has the structure shown in Formula 12; the fucose acceptor 11 has the structure shown in Formula 240; the first glycosylation product disaccharide 28 has the structure shown in Formula 28; the disaccharide acceptor 29 has the structure shown in Formula 29; the fucose PTFAI donor 21d has the structure shown in Formula 21d; the trisaccharide 30 has the structure shown in Formula 30; the trisaccharide PTFAI donor 9 has the structure shown in Formula 9; the hexasaccharide 10 has the structure shown in Formula 10; and the nonasugar 31 has the structure shown in Formula 31. Equation 12; Formula 24o; Equation 28; Equation 29; Equation 21d; Formula 30; Equation 9; Formula 10; Equation 31; The arabinose ABz receptor 14 has the structure shown in Formula 14; the arabinose PVB receptor 15 has the structure shown in Formula 15; the arabinose receptor 16 has the structure shown in Formula 16. Formula 14; Formula 15; Formula 16.

2. The production method according to claim 1, characterized by, The preparation of the hexasaccharide receptor 10 includes the following steps: The arabinose PTFAI donor 18, glucose PVB acceptor 19, first activator, fourth dehydrating agent and fifth polar organic solvent were mixed and subjected to the fourth glycosylation reaction to obtain a reaction solution containing α-Araf-(1→6)-Glc disaccharide. The reaction solution containing α-Araf-(1→6)-Glc disaccharide, arabinose acceptor 20, second activator, and N,N-dimethylformamide were mixed and subjected to a fifth glycosylation reaction to obtain trisaccharide 26; the protecting group MP of the trisaccharide 26 was removed to obtain a hemiacetal intermediate; the hemiacetal intermediate, a sixth polar organic solvent, 2,2,2-trifluoro-N-phenyliminoacetyl chloride, and a second carbonate were mixed and subjected to a second substitution reaction to obtain trisaccharide PTFAI donor 17; The trisaccharide PTFAI donor 17, arabinose ABz acceptor 14, fifth dehydrating agent, seventh polar organic solvent and third activator are mixed and subjected to a sixth glycosylation reaction to obtain a reaction solution containing a tetrasaccharide ABz intermediate; the reaction solution containing the tetrasaccharide ABz intermediate, arabinose PVB acceptor 15 and catalyst are mixed and subjected to a seventh glycosylation reaction to obtain a reaction solution containing a pentasaccharide intermediate; The reaction solution containing the pentasaccharide intermediate, arabinose acceptor 16, and fourth activator were mixed and subjected to an eighth glycosylation reaction to obtain hexasaccharide 27; the hexasaccharide 27 was then selectively deprotected by the TIPS protecting group to obtain hexasaccharide 10. The arabinose PTFAI donor 18 has the structure shown in Formula 18; the glucose PVB receptor 19 has the structure shown in Formula 19; the arabinose receptor 20 has the structure shown in Formula 20; the trisaccharide 26 has the structure shown in Formula 26; and the trisaccharide PTFAI donor 17 has the structure shown in Formula 17. Formula 18; Formula 20; Formula 26; Formula 17.

3. A method for preparing a 13-saccharide repeating unit of Angelica polysaccharide APS-1 II, wherein the 13-saccharide repeating unit has the structure shown in Formula 1; Formula 1; The preparation method Includes the following steps: The preparation method according to claim 1 or 2 yields a nonaglycone 31; the Lev protecting group of the nonaglycone 31 is removed to obtain a nonaglycone 8; a glucose PTFAI donor 5, a glucose ABZ acceptor 6, a dehydrating agent, a first polar organic solvent, and a first activator are mixed to perform a ninth glycosylation reaction to obtain a reaction solution containing a disaccharide ABZ intermediate; the reaction solution containing the disaccharide ABZ intermediate, an arabinose PVB acceptor 7, and a catalyst are mixed to perform a tenth glycosylation reaction to obtain a reaction solution containing a tetrasaccharide PVB intermediate; the reaction solution containing the tetrasaccharide PVB intermediate, the nonaglycone 8, and a second activator are mixed to perform an eleventh glycosylation reaction to obtain a decaglycone 35; The protecting group of the decadecose 35 was removed to obtain the 13-saccharide repeating unit of the Angelica polysaccharide APS-1 II; The nonaglycone 8 has the structure shown in Formula 8; the glucose PTFAI donor 5 has the structure shown in Formula 5; the glucose ABZ receptor 6 has the structure shown in Formula 6; the arabinose PVB receptor 7 has the structure shown in Formula 7; and the tridecanose 35 has the structure shown in Formula 35. Formula 8; Formula 35; Formula 5; Formula 6; Formula 7.

4. Angelica sinensis polysaccharide APS-1 II, characterized in that, The Angelica polysaccharide APS-1 II has a substructure 9 sugar prepared by the preparation method of claim 1 or 2 and a 13 sugar repeating unit prepared by the preparation method of claim 3; the 13 sugar repeating unit has the structure shown in Formula 1, and the substructure 9 sugar has the structure shown in Formula 2. Formula 1 ; Formula 2.

5. The use of Angelica polysaccharide APS-1 II as described in claim 4 in the preparation of a drug for inhibiting the proliferation of leukemia cells.

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