Method for stereospecifically synthesizing Tn antigen and TF antigen

Abstract

The invention discloses a method for stereospecifically synthesizing a Tn antigen and a TF antigen, and belongs to the technical field of synthesis of the Tn antigen and the TF antigen, and the specific method comprises the following steps: taking 2-azido-2-deoxy-3-O-tertiary butyl dimethyl silyl-4, 6-di-O-benzoyl-galactosyl phenyl thioester as a donor, taking serine as a receptor, and carrying out a one-pot reaction to obtain the Tn antigen and the TF antigen. Adopting a remote adjacent group to participate in effect stereospecificity to synthesize a protected Tn antigen; selectively removing tert-butyldimethylsilyl of the Tn antigen to dissociate 3-site hydroxyl, and reacting with peracetylated galactosyl trichloroacetimidate to obtain a protected TF antigen; and finally, removing protecting groups for protecting the Tn antigen and the TF antigen to obtain the Tn antigen and the TF antigen. The synthesis method has the advantages of mild conditions, low cost, high yield and the like, and meanwhile, the remote participation effect can be enhanced, and the stereospecificity can be efficiently realized.

Description

Technical Field

[0001] The invention belongs to the technical field of synthesis of Tn antigen and TF antigen, and particularly relates to a method for synthesizing Tn antigen and TF antigen with stereospecificity. Background Art

[0002] Tn antigen is a monosaccharide amino acid molecule that is highly expressed in a variety of cancer cells, such as breast cancer, lung cancer, and colorectal cancer. Therefore, this Tn antigen is used as a biomarker and target for tumor vaccine therapy. Many tumor-associated carbohydrate antigens (TACAs) have the structure of α-GalNAc-Ser, such as TF antigen, T antigen, and Sialyl-T antigen, which are targets for the development of new cancer treatment and diagnostic tools. Since it is difficult to obtain a large amount of these molecules in a uniform form from natural resources, it is particularly meaningful to develop highly selective reaction schemes for these structures. In many cases, high-purity Tn antigen can only be obtained by chemical or enzymatic synthesis. There is a very difficult problem in the chemical synthesis of Tn antigen: the stereospecific construction of the α-GalNAc glycosidic bond. The key to the preparation of TF antigen is the directional connection of galactose to the 3-hydroxyl group of acetylgalactosamine. Nowadays, most of the chemically synthesized Tn antigens connect the 6th position of the donor to -TBDPS, and the 3rd and 4th positions to acyl groups, and high stereoselectivity can be achieved by using the remote participation effect, such as the work done by George Ngoje et al. (J.Org Biomol Chem.2011,9,6825). In actual work, we repeated their work and found that when the milligram-level reaction was performed, the stereoselectivity reached 15:1 as described, but when the gram-level reaction was performed, the selectivity decreased. Moreover, if the group design of the above-mentioned document is used, it is very difficult to free the 3rd hydroxyl group. Therefore, finding a method that can achieve both the stereospecificity of the α-GalNAc glycosidic bond and the selective freeing of the 3rd hydroxyl group of acetylgalactosamine has become a technical problem that needs to be solved in the art. Summary of the invention

[0003] In view of the above-mentioned deficiencies in the prior art, the present invention provides a method for stereospecifically synthesizing Tn antigen and TF antigen, which has the advantages of mild conditions, low cost, high yield, etc., and can also enhance the remote participation effect and efficiently achieve stereospecificity.

[0004] To achieve the above purpose, the technical solution adopted by the present invention to solve the technical problem is:

[0005] A method for stereospecifically synthesizing Tn antigen and TF antigen comprises the following steps:

[0006] S1: Using 3,4,6-tri-O-acetyl-galactenose as the initial substrate, compound 1 is subjected to an addition reaction at the 1,2-position of compound 1 to obtain compound 2.

[0007]

[0008] S2: Replace -ONO in compound 2 with Br 2 The group is then reacted with thiophenol to obtain compound 3.

[0009]

[0010] S3: remove all acetyl groups of compound 3 to obtain compound 4,

[0011]

[0012] S4: Protect the 4- and 6-hydroxyl groups of compound 4 with acetal, and replace the 3-hydroxyl group with -OTBS group to obtain compound 5.

[0013]

[0014] S5: removing the acetal protecting group of compound 5 and replacing the 4,6-hydroxyl groups with -OBz ​​groups to obtain compound 6.

[0015]

[0016] S6: Protect the amino group and carboxyl group of L-serine with -Cbz and -Bn respectively to obtain compound 7.

[0017]

[0018] S7: Compound 6 and compound 7 are subjected to glycosylation reaction to obtain compound 8.

[0019]

[0020] S8: removing the -TBS group of compound 8 to obtain compound 9,

[0021]

[0022] S9: Compound 9 is subjected to a glycosylation reaction with 2,3,4,6-tetra-O-acetyl-α / β-D-galactopyranose trichloroacetimidate to obtain compound 10.

[0023]

[0024] S10: Compound 9 and compound 10 are subjected to group treatment to obtain compound 11 and compound 12

[0025]

[0026] Furthermore, the specific preparation process is as follows:

[0027] S1: Compound 1 was dissolved in dry acetonitrile, and then cerium ammonium nitrate (CAN) was added. The system was stirred at 0°C for 10 minutes, and then trimethylsilyl azide (TMSN) was added dropwise. 3 ), react in an ice bath overnight, add ice water until the solid dissolves, extract with ethyl acetate, dry, concentrate, and purify by column chromatography to obtain compound 2;

[0028] S2: Compound 2 was dissolved in dry acetonitrile, and the mixture was reacted at 0°C for 2 hours. Lithium bromide was then added, and the mixture was heated to room temperature for 4-6 hours. The mixture was extracted with dichloromethane, dried, and concentrated to obtain product 1. Product 1 was dissolved in dry dichloromethane, and the mixture was stirred at -20°C for 10 minutes. Thiophenol was added and stirred to mix well. Boron trifluoride ether solution was slowly added dropwise. 2 The mixture was reacted for 20 minutes under protection, triethylamine was added to adjust the pH to 7-8, and the mixture was extracted with dichloromethane, dried, concentrated, and purified by column chromatography to obtain compound 3;

[0029] S3: Compound 3 was mixed with 0.3 mmol / mL MeONa / MeOH solution, stirred at room temperature for 30 minutes, concentrated, and purified by column chromatography to obtain compound 4;

[0030] S4: Compound 4 was dissolved in dry acetonitrile, and then benzaldehyde dimethyl acetal and p-toluenesulfonic acid were added in sequence, and the mixture was allowed to react at room temperature for 3 hours, and triethylamine was added to adjust the pH to 7-8, and the mixture was concentrated, extracted with dichloromethane, dried, and concentrated to obtain intermediate 1; intermediate 1 was dissolved in dry dichloromethane, stirred at room temperature, and activated imidazole and tert-butyldimethylsilyl chloride were added in sequence, and the mixture was allowed to react at room temperature overnight, extracted with dichloromethane, dried, concentrated, and purified by column chromatography to obtain compound 5;

[0031] S5: Compound 5 was mixed with 80% acetic acid aqueous solution and stirred under N 2 Under protection, the mixture was placed under condensation reflux at 80°C for 20 minutes, concentrated, and purified by column chromatography to obtain product II; product II was dissolved in anhydrous pyridine, stirred at 0°C for 5 minutes, benzoyl chloride was slowly added dropwise, the temperature was raised to 40°C for reaction for 2 hours, methanol was added to quench, dichloromethane was extracted, and the aqueous layer was washed with 1N dilute HCl until it became acidic, dried, concentrated, and purified by column chromatography to obtain compound 6;

[0032] S6: Add L-serine in batches to a sodium bicarbonate aqueous solution precooled at 0°C, cool to 0°C, slowly drop a 50% solution of benzyl chloroformate and toluene, continue stirring at 0°C for 2h, warm to room temperature and stir for 2h, add ethyl acetate, adjust the pH to 2.0 with 1N dilute hydrochloric acid, extract with ethyl acetate, and concentrate until one-third of the liquid remains to obtain a suspension, add cyclohexane and stir for 6h, filter and dry to obtain product three; under nitrogen protection, dissolve product three in dry N,N-dimethylformamide, add benzyl bromide and cesium carbonate in sequence, stir at room temperature overnight, quench with water, extract with dichloromethane, concentrate, and purify by column chromatography to obtain compound 7;

[0033] S7: Compound 6 and Compound 7 were dissolved in dry dichloromethane and added molecular sieves, stirred at 0°C for 5-10 minutes, then N-iodosuccinimide was added in an inert gas atmosphere, stirred at 0°C for 5-10 minutes, then trifluoromethanesulfonic acid was added, the temperature was raised to room temperature and reacted for 2 hours, then triethylamine was added to adjust the pH to 7-8, concentrated, and purified by column chromatography to obtain compound 8;

[0034] S8: Compound 8 was dissolved in dry dichloromethane, and a boron trifluoride ether solution was added dropwise under an ice bath, and the mixture was heated to room temperature and stirred. The reaction was allowed to proceed overnight, and triethylamine was added to adjust the pH to 7-8. The mixture was concentrated and purified by column chromatography to obtain compound 9;

[0035] S9: Compound 9 and 2,3,4,6-tetra-O-acetyl-α / β-D-galactopyranose trichloroacetimidate were dissolved in dry dichloromethane and added molecular sieves, stirred at 0°C for 5-10 minutes, then trimethylsilyl trifluoromethanesulfonate was added in an inert gas atmosphere, and the reaction was continued at 0°C for 2 hours, and then triethylamine was added to adjust the pH to 7-8, concentrated, and purified by column chromatography to obtain compound 10;

[0036] S10: Under nitrogen protection, compound 9 was dissolved in pyridine, and then thioacetic acid was added dropwise, and the reaction was stirred at room temperature overnight. After the reaction was completed by thin layer chromatography, the reaction solution was concentrated and purified by column chromatography to obtain intermediate 2; intermediate 2 was dissolved in methanol, and then water, glacial acetic acid, and 10% palladium carbon were added in sequence, and hydrogen was introduced into the reaction system. The reaction was reacted at room temperature for 6 hours. After the reaction was completed by thin layer chromatography, the reaction solution was filtered and the filtrate was concentrated to obtain a white solid; under nitrogen protection, the above white solid was dissolved in methanol, and then 1,4-dioxane and 1M sodium hydroxide aqueous solution were added in sequence. The reaction was reacted at room temperature overnight. After the reaction was completed by thin layer chromatography, the pH was adjusted to 7.0 with glacial acetic acid, and the reaction solution was concentrated and purified by Sephadex LH-20 gel column chromatography to obtain compound 11; compound 10 was subjected to the above operation steps to obtain compound 12;

[0037] The above reaction process is as follows:

[0038]

[0039] Further, in S1, the concentration of compound 1 in acetonitrile is 0.1-0.3 mmol / mL, and the molar ratio of compound 1, ammonium cerium nitrate and trimethylsilyl azide is 0.8-1.2:1.5-2.5:1.5-2;

[0040] In S2, the concentration of compound 2 in acetonitrile is 0.1-0.3 mmol / mL, and the molar ratio of compound 2 to lithium bromide is 1:4-6; the concentration of product 1 in dichloromethane is 0.1-0.3 mmol / mL, and the molar ratio of product 1, thiophenol and boron trifluoride ether is 1:1.3-2.2:1.3-2.3;

[0041] The concentration of compound 3 in MeONa / MeOH in S3 is 0.3-0.5 mmol / mL; the reaction time is 20-40 min;

[0042] In S4, the concentration of compound 4 in acetonitrile is 0.1-0.3 mmol / mL, and the molar ratio of compound 4, benzaldehyde dimethyl acetal and p-toluenesulfonic acid is 1:1.3-1.6:0.06-0.1; the concentration of intermediate 1 in dichloromethane is 0.2-0.5 mmol / mL, and the molar ratio of intermediate 1, imidazole and tert-butyldimethylchlorosilane is 1:2.1-2.7:2.1-2.7;

[0043] The molar ratio of compound 5 to acetic acid in S5 is 1:90-110, the concentration of product II in pyridine is 0.4-0.6 mmol / mL, and the molar ratio of product II to benzoyl chloride is 1:7-9;

[0044] The concentration of the sodium bicarbonate aqueous solution in S6 is 1.8-2.0 mmol / mL, the molar ratio of L-serine, sodium bicarbonate and benzyl chloroformate is 1:2.5-3.5:1-1.5, the concentration of the solution prepared by the product three and N,N-dimethylformamide is 0.1-0.3 mmol / mL, and the molar ratio of the product three, benzyl bromide and cesium carbonate is 1:1-1.5:1-1.5;

[0045] In S7, the concentration of compound 7 in dichloromethane is 0.04-0.06 mmol / mL, and the molar ratio of compound 6, compound 7, N-iodosuccinimide and trifluoromethanesulfonic acid is 1.2-1.5:1:1-1.4:0.05-0.15;

[0046] In S8, the concentration of compound 8 in dichloromethane is 0.3-0.5 mmol / mL, and the molar ratio of compound 8 to boron trifluoride ether is 1:2-4;

[0047] In S9, the concentration of compound 9 in dichloromethane is 0.04-0.06 mmol / mL, and the molar ratio of compound 9, 2,3,4,6-tetra-O-acetyl-α / β-D-galactopyranose trichloroacetimidate and trimethylsilyl trifluoromethanesulfonate is 1:1.4-1.6:0.05-0.15;

[0048] The concentration of compound 9 in S10 in the pyridine solution is 0.04-0.06 mmol / mL, and the molar ratio of compound 9 to thioacetic acid is 1:170-190; the molar volume ratio of intermediate 2, methanol, water, and glacial acetic acid is 1 mmol:25-45 mL:2.5-4.5 mL:250-450 μL, and the molar ratio of intermediate 2 to palladium carbon is 1:1.5-2.5; the molar volume ratio of white solid, methanol, 1,4-dioxane, and 1M sodium hydroxide aqueous solution is 1 mmol:8-12 mL:8-12 mL:12-18 mL;

[0049] The concentration of compound 10 in S10 in pyridine solution is 0.04-0.06 mmol / mL, and the molar ratio of compound 10 to thioacetic acid is 1:170-190; the molar volume ratio of intermediate three, methanol, water, and glacial acetic acid is 1 mmol:25-45 mL:2.5-4.5 mL:250-450 μL, and the molar ratio of intermediate three to palladium carbon is 1:1.5-2.5; the molar volume ratio of white solid, methanol, 1,4-dioxane, and 1M sodium hydroxide aqueous solution is 1 mmol:8-12 mL:8-12 mL:12-18 mL.

[0050] The beneficial effects produced by the present invention are:

[0051] In the present invention, a -TBS group is selected at the 3rd position of the donor, and an acyl group is selected at the 4th and 6th positions. -TBS is a strong electron-donating group, which can not only achieve the effect of strengthening the remote participation effect, but also can be easily removed to selectively free the 3rd hydroxyl group, laying the foundation for the stereospecific connection between the TF antigen and the acetylated galactose. Moreover, the glycosylation conditions used in the present invention are mild, the experimental operation is simple, the cost is low, and the yield is relatively high, which can make the glycosylation reaction yield reach 82%-85%. In the case of gram-level reaction, the selectivity can still reach α:β greater than 20:1, the reaction is efficient and its stereospecificity is achieved. DETAILED DESCRIPTION

[0052] In order to make the purpose, technical scheme and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention, that is, the embodiments described are only part of the embodiments of the present invention, rather than all of the embodiments.

[0053] Therefore, the following detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present invention.

[0054] It should be noted that relational terms such as "first" and "second" are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprises" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, an element defined by the sentence "comprises a ..." does not exclude the existence of other identical elements in the process, method, article or device including the element.

[0055] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0056] Example 1

[0057] A method for stereospecifically synthesizing Tn antigen and TF antigen comprises the following steps:

[0058] S1: Compound 2 is obtained by addition reaction at the 1,2-position of compound 1. The reaction equation is as follows:

[0059]

[0060] The specific steps are as follows: Compound 1 (12.0 g, 44.1 mmol) was dissolved in dry acetonitrile (220 mL, 0.2 M), the system was cooled to -15 °C, ammonium cerium nitrate (46.60 g, 85.0 mmol) was added and stirred for 5 min, trimethylsilane (8.7 mL, 66.0 mmol) was added, and N 2 The temperature was raised to 0°C under protection and the reaction was allowed to proceed overnight. After the reaction was complete, ice water was added to the system to quench until the solid was completely dissolved, extracted with ethyl acetate, and concentrated under reduced pressure in vacuo. The crude product was separated and purified by chromatography on a silica gel column (5:1, PE:EA) to obtain compound 2 (6.63 g, 40%).

[0061] S2: -ONO in compound 2 2After the group is replaced by a halogen atom Br, it is glycosylated with thiophenol to obtain compound 3. The reaction equation is as follows:

[0062]

[0063] The specific steps are as follows: Compound 2 (6.63 g, 17.64 mmol) was dissolved in dry acetonitrile (88 mL, 0.2 M), reacted under ice bath for 2 h, and then lithium bromide (7.66 g, 88.20 mmol) was added. 2 The reaction mixture was heated to room temperature under protection for 4-6 hours. After the reaction was complete, the mixture was extracted with dichloromethane and concentrated under vacuum to obtain product 1 (6.90 g, 16.86 mmol). Product 1 was dissolved in dry dichloromethane (84 mL, 0.2 M), the system was cooled to -20 °C, thiophenol (3.5 mL, 34.50 mmol) was added and stirred for 5 minutes, and then boron trifluoride ether solution (2.5 mL, 24.30 mmol) was added. 2 The mixture was reacted at -20°C for 30 min under protection. After the reaction was complete, the pH was adjusted to 7-8 with triethylamine, extracted with dichloromethane, and concentrated under vacuum. The crude product was separated and purified by chromatography on a silica gel column (6:1, PE:EA) to obtain compound 3 (4.20 g, 59%).

[0064] S3: Remove the acetyl group of compound 3 to obtain compound 4. The reaction equation is as follows:

[0065]

[0066] The specific steps are as follows: sodium methoxide is dissolved in methanol to prepare a 0.3M sodium methoxide / methanol solution, compound 3 (4.20 g, 9.92 mmol) and 0.3M sodium methoxide / methanol solution (24.8 mL, 0.4 M) are mixed and stirred for 30 min, and after the reaction is complete, the mixture is concentrated under vacuum, and the crude product is separated and purified by chromatography on a silica gel column (20:1, DCM:MeOH) to obtain compound 4 (2.34 g, 80%);

[0067] S4: The 4- and 6-hydroxyl groups of compound 4 are protected with acetal, and the 3-hydroxyl group thereof is replaced with an -OTBS group to obtain compound 5. The reaction equation is as follows:

[0068]

[0069] The specific steps are as follows: Compound 4 (2.34 g, 7.94 mmol) was dissolved in dry acetonitrile (40 mL, 0.2 M), benzaldehyde dimethyl acetal (1.7 mL, 11.50 mmol) was added, and p-toluenesulfonic acid monohydrate (121 mg, 0.64 mmol) was added. 2The mixture was reacted at room temperature for 3 h under protection. After the reaction was complete, triethylamine was added to adjust the pH to 7-8, and the mixture was extracted with dichloromethane and concentrated under vacuum to obtain intermediate 1 (2.23 g, 5.79 mmol). Intermediate 1 was dissolved in dry dichloromethane (16.5 mL, 0.35 M), activated imidazole (945 mg, 13.90 mmol) was added, and tert-butyldimethylsilyl chloride (2.10 g, 13.90 mmol) was added. 2 The reaction was allowed to proceed overnight at room temperature under protection. After the reaction was complete, the mixture was extracted with dichloromethane and concentrated under vacuum. The crude product was separated and purified by chromatography on a silica gel column (50:1, PE:EA) to obtain compound 5 (2.34 g, 81%).

[0070] S5: The acetal protecting group of compound 5 is removed, and the 4- and 6-hydroxyl groups are replaced with -OBz ​​groups to obtain compound 6. The reaction equation is as follows:

[0071]

[0072] The specific steps are as follows: acetic acid (26.7 mL, 468 mmol) and water (6.7 mL) are prepared into an 80% acetic acid aqueous solution, compound 5 (2.34 g, 4.68 mmol) is mixed therewith, N 2 The reaction mixture was stirred at 80°C for 20 min under protection. After the reaction was complete, the mixture was concentrated under vacuum to an oily substance. The crude product was separated and purified by chromatography on a silica gel column (8:1, PE:EA) to obtain product II (1.58 g, 3.84 mmol). Product II was dissolved in anhydrous pyridine (7.7 mL, 0.5 M), cooled to 0°C, benzoyl chloride (3.6 mL, 30 mmol) was added dropwise, the temperature was raised to room temperature, and N 2 The reaction was carried out under protection for 2 h. After the reaction was complete, methanol was added to quench the reaction, and the mixture was extracted with dichloromethane. The mixture was washed with 1N dilute hydrochloric acid, saturated sodium bicarbonate solution, and saturated brine. After vacuum concentration, the crude product was separated and purified by chromatography on a silica gel column (30:1, PE:EA) to obtain compound 6 (2.21 g, 93%).

[0073] S6: The amino group and carboxyl group of L-serine were protected with -Cbz and -Bn, respectively, to obtain compound 7. The reaction equation is as follows:

[0074]

[0075] The specific steps are as follows: sodium bicarbonate (2.4 g, 28.56 mmol) was dissolved in water (15 mL) and precooled at 0°C, L-serine (1 g, 9.52 mmol) was added in batches, stirred for 5 min, benzyl chloroformate (1.6 mL, 11.42 mmol) and toluene (1.6 mL) were prepared into a 50% solution, and the solution was slowly added dropwise into the system, and N2 The mixture was stirred at 0°C for 2 h under protection, then heated to room temperature and stirred for 2 h. After the reaction was complete, ethyl acetate (50 mL) was added, and the pH was adjusted to 2.0 with 1N dilute hydrochloric acid. The mixture was extracted with ethyl acetate and concentrated under vacuum until one third of the liquid remained. Cyclohexane (100 mL) was added and stirred for 6 h. The mixture was filtered and dried to obtain the product III (1.20 g, 5 mmol). The product III was dissolved in N,N-dimethylformamide (25 mL, 0.2 M), and benzyl bromide (0.6 mL, 5 mmol), cesium carbonate (1.63 g, 5 mmol), N 2 The reaction was allowed to proceed overnight at room temperature under protection. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with dichloromethane and concentrated under vacuum. The crude product was separated and purified by chromatography on a silica gel column (2:1, PE:EA) to obtain compound 7 (1.28 g, 78%).

[0076] S7: Compound 6 and compound 7 are subjected to glycosylation reaction to obtain compound 8. The reaction equation is as follows:

[0077]

[0078] The specific steps are as follows: Compound 6 (1.70 g, 2.74 mmol) was dissolved in dichloromethane (45 mL, 0.05 M), and then compound 7 (751 mg, 2.28 mmol) was added. The molecular sieves were stirred at 0°C for 10 min, and then 2 N-iodosuccinimide (740 mg, 3.29 mmol) was added under an aqueous solution of 4,4-dichloromethane (2,4-dichloroethane) and stirred at 0°C for 5 min. Then trifluoromethanesulfonic acid (20 μL, 0.23 mmol) was slowly added. After the addition, the temperature was raised to room temperature and the reaction was continued for 2 h. After the reaction was completed, triethylamine was added to quench the reaction. The mixture was extracted with dichloromethane and concentrated under vacuum. The crude product was separated and purified by chromatography on a silica gel column (20:1, PE:EA) to obtain compound 8 (1.63 g, 85%).

[0079] 1 H NMR (600 MHz, CDCl 3)δ8.04–8.02(m,2H),8.01–7.99(m,2H),7.59(t,J=7.2Hz,1H),7.53(t,J= 7.2Hz,1H),7.46(t,J=7.8Hz,2H),7.40(d,J=7.8Hz,2H),7.36(s,5H),7.31 (dd,J=6.3,3.9Hz,5H),5.83(d,J=8.4Hz,1H),5.56(d,J=3.6Hz,1H),5.22( d,J=2.4Hz,2H),5.05(s,2H),4.97(d,J=3.6Hz,1H),4.63(d,J=8.4Hz,1H), 4.43–4.38 (m, 1H), 4.30–4.26 (m, 2H), 4.13 (dd, J = 10.2, 3.3 Hz, 1H), 4.09 (dd, J = 10.8, 3.3 Hz, 1H), 4.03 (dd, J = 10.8, 3.0 Hz, 1H), 3.64 (dd, J = 10.2, 3.6 Hz, 1H), 0.77 (s, 9H), 0.17 (d, J = 6.6 Hz, 6H). δ4.97 (d, J = 3.6 Hz, 1H) is the hydrogen of the anomeric carbon of the sugar ring, and the coupling constant is 3.6, which belongs to the α-H category. When the reaction is at the hundred milligram level, it is α-only; when the reaction is at the nearly 2 gram level, the ratio of its peak area to the isomer is greater than 20:1.

[0080] 13 C NMR (151 MHz, CDCl 3 )δ169.72,166.26,165.78,156.01,136.19,135.20,133.44,133.28,129.94,129.87,129.72,128.80,128.73,128.69,128.63,128 .56,128.41,128.30,99.26,70.94,69.60,68.26,67.88,67.31,63.37,61.87,54.63,25.65,17.86,-4.44,-5.00.HRMS(ESI)calcd for C 44 H 50 N 4 NaO 11 Si[M+Na] + 861.3143, found 861.3146.

[0081] S8: The -TBS group of compound 8 is removed to obtain compound 9. The reaction equation is as follows:

[0082]

[0083] The specific steps are as follows: Compound 8 (1.63 g, 1.94 mmol) was dissolved in dry dichloromethane (5 mL, 0.4 M), cooled to 0°C, and then a boron trifluoride ether solution (670 μL, 5.82 mmol) was added dropwise, and the mixture was heated to room temperature and stirred. The mixture was reacted overnight, and triethylamine was added to quench the reaction. The mixture was concentrated under reduced pressure in vacuo. The crude product was separated and purified by chromatography on a silica gel column (3:1, PE:EA) to obtain Compound 9 (1.10 g, 78%).

[0084] S9: Compound 9 is subjected to a glycosylation reaction with 2,3,4,6-tetra-O-acetyl-α / β-D-galactopyranose trichloroacetimidate to obtain compound 10. The reaction equation is as follows:

[0085]

[0086] The specific steps are as follows: Compound 9 (500 mg, 0.690 mmol) was dissolved in dichloromethane (14 mL, 0.05 M), and then compound 2,3,4,6-tetra-O-acetyl-α / β-D-pyranose trichloroacetimidate (509 mg, 1.04 mmol) was added. The molecular sieves were stirred at 0°C for 10 min, and then 2 Trimethylsilyl trifluoromethanesulfonate (13 μL, 0.069 mmol) was slowly added under an atmosphere, and the mixture was kept warm (0°C) for 2 h. After the reaction was completed, triethylamine was added to quench the reaction. After vacuum concentration, the crude product was separated and purified by chromatography on a silica gel column (2:1, PE:EA) to obtain compound 10 (597 mg, 82%).

[0087] 1 H NMR (600 MHz, CDCl 3)δ8.02(d,J=7.2Hz,2H),7.98(d,J=6.9Hz,2H),7.58(t,J=7.5Hz,1H),7.54–7.51(m,1H),7. 48–7.45(m,2H),7.38(dd,J=10.8,6.3Hz,7H),7.34–7.29(m,5H),5.77(d,J=8.4Hz,1H),5.7 4(d,J=3.6Hz,1H),5.29(d,J=3.9Hz,1H),5.22(d,J=2.7Hz,2H),5.12–5.06(m,3H),4.97(dd ,J=10.2,3.3Hz,1H),4.93(d,J=3.6Hz,1H),4.72(d,J=7.8Hz,1H),4.65–4.60(m,1H),4.40( dd,J=11.7,4.2Hz,1H),4.33(dd,J=11.7,7.8Hz,1H),4.25(dd,J=8.7,4.2Hz,1H),4.15(dd, J=10.8,3.6Hz,1H),4.10(dd,J=11.4,6.3Hz,1H),4.05(dd,J=10.5,3.3Hz,1H),4.02–3.96( m, 2H), 3.90 (t, J = 6.6 Hz, 1H), 3.72 (dd, J = 10.8, 3.6 Hz, 1H), 2.03 (s, 3H), 2.00 (s, 3H), 1.94 (s, 3H), 1.91 (s, 3H). δ4.72 (d, J = 7.8 Hz, 1H) is the hydrogen of the anomeric carbon of the galactose ring, and the coupling constant is 7.8, which belongs to the category of β-H, so it is determined that the main product is β configuration.

[0088] 13 C NMR (151 MHz, CDCl 3 )δ170.56,170.31,170.24,169.69,169.52,165.69,155.98,136.11,135.16,133.36,1 33.28,130.12,129.85,129.82,129.71,128.84,128.79,128.73,128.63,128.57,128. 55,128.49,128.34,101.62,99.08,70.98,70.93,70.33,69.36,68.89,68.30,67.91,6 7.40,66.82,63.51,61.24,59.44,54.51,20.83,20.77,20.67,20.51.HRMS(ESI)calcd for C 52 H55 N 4 O 20 [M+H] + 1055.3410, found 1055.3422.

[0089] S10: Compound 9 is subjected to group treatment to obtain compound 11 (Tn antigen). The reaction equation is as follows:

[0090]

[0091] The specific steps are as follows: 2 Under a nitrogen atmosphere, compound 9 (300 mg, 0.414 mmol) was dissolved in pyridine (7.4 mL, 0.056 M), and then thioacetic acid (5.3 mL, 75 mmol) was added dropwise. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by thin layer chromatography, the mixture was extracted with ethyl acetate and concentrated to obtain intermediate 2. Intermediate 2 was dissolved in methanol (15 mL), and then water (1.5 mL), glacial acetic acid (150 μL), and 10% palladium carbon (890 mg, 0.83 mmol) were added in sequence. Hydrogen was introduced into the reaction system, and the reaction was allowed to react for 6 h at room temperature. After the reaction was completed as monitored by thin layer chromatography, the reaction solution was filtered, and the filtrate was concentrated to obtain a white solid. Under nitrogen protection, the above white solid was dissolved in methanol (4.1 mL), and then 1,4-dioxane (4.1 mL) and 1M sodium hydroxide aqueous solution (6.2 mL) were added in sequence. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by thin layer chromatography, the pH was adjusted to 7.0 with glacial acetic acid, and the reaction solution was concentrated and Sephadex was used. LH-20 gel column chromatography purification (1:1, H 2 O: MeOH), to obtain compound 11 (Tn antigen) (42 mg, 33%);

[0092] 1 H NMR (600MHz, D 2 O) δ4.92(d,J=3.9Hz,1H),4.21(dd,J=11.1,3.9Hz,1H),4.13(dd,J=11.1,3.0Hz,1H),3.83–3.73(m,2H),2.05(s,3H).

[0093] 13 C NMR (101MHz, D 2 O)δ174.4,171.3,97.5,71.1,68.1,67.3,66.3,60.9,54.2,49.3,21.5.HRMS(ESI)calcd for C 11 H 19 N 2 O 8 [MH]- 307.1147, found 307.1148.

[0094] S11: Compound 10 is subjected to group treatment to obtain compound 12 (TF antigen). The reaction equation is as follows:

[0095]

[0096] The specific steps are as follows: 2 Under an argon atmosphere, compound 10 (280 mg, 0.266 mmol) was dissolved in pyridine (5 mL, 0.056 M), and then thioacetic acid (3.4 mL, 47.7 mmol) was added dropwise. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by thin layer chromatography, the mixture was extracted with ethyl acetate and concentrated to obtain intermediate 3. Intermediate 3 was dissolved in methanol (9.5 mL), and then water (950 μL), glacial acetic acid (100 μL), 10% palladium carbon (570 mg, 0.53 2mmol), hydrogen was introduced into the reaction system, and the reaction was carried out at room temperature for 6h. After the reaction was completed as monitored by thin layer chromatography, the reaction solution was filtered and the filtrate was concentrated to obtain a white solid. Under nitrogen protection, the white solid was dissolved in methanol (2.7mL), and 1,4-dioxane (2.7mL) and 1M sodium hydroxide aqueous solution (4mL) were added in sequence, and the reaction was carried out at room temperature overnight. After the reaction was completed as monitored by thin layer chromatography, glacial acetic acid was used to adjust the pH to 7.0, and the reaction solution was concentrated and purified by Sephadex LH-20 gel column chromatography (1:1, H 2 O: MeOH), to obtain compound 12 (TF antigen) (38 mg, 30%);

[0097] 1 H NMR (600MHz, D 2 O)δ4.92(d,J=3.3Hz,1H),4.47(d,J=7.8Hz,1H),4.35(dd,J=11.1,3.3Hz,1H),4.23(s,1H),4.17–4.02(m,2H) ,4.04–3.95(m,2H),3.96–3.87(m,2H),3.84–3.72(m,4H),3.67–3.60(m,2H),3.55–3.49(m,1H),2.02(s,3H).

[0098] 13 C NMR (101MHz, D 2 O)δ174.7,171.9,104.7,98.5,76.8,75.3,72.6,71.3,70.6,68.7,68.7,61.4,61.1,48.5,22.2.HRMS(ESI)calcd for C17 H 29 N 2 O 13 [MH] - 469.1675, found 469.1678. Example 2

[0099] A method for stereospecifically synthesizing Tn antigen and TF antigen comprises the following steps:

[0100] S1: Compound 2 is obtained by addition reaction at the 1,2-position of compound 1. The reaction equation is as follows:

[0101]

[0102] The specific steps are as follows: Compound 1 (12.0 g, 44.1 mmol) was dissolved in dry acetonitrile (440 mL, 0.1 M), the system was cooled to -15 °C, ammonium cerium nitrate (45.33 g, 82.69 mmol) was added and stirred for 5 min, trimethylsilane (10.6 mL, 82.69 mmol) was added, and N 2 The temperature was raised to 0°C under protection and the reaction was allowed to proceed overnight. After the reaction was complete, ice water was added to the system to quench until the solid was completely dissolved, extracted with ethyl acetate, and concentrated under reduced pressure in vacuo. The crude product was separated and purified by chromatography on a silica gel column (5:1, PE:EA) to obtain compound 2 (6.2 g, 37%).

[0103] S2: -ONO in compound 2 2 After the group is replaced by a halogen atom Br, it is glycosylated with thiophenol to obtain compound 3. The reaction equation is as follows:

[0104]

[0105] The specific steps are as follows: Compound 2 (6.2 g, 16.48 mmol) was dissolved in dry acetonitrile (165 mL, 0.1 M), reacted under ice bath for 2 h, and then lithium bromide (5.73 g, 65.92 mmol) was added. 2 The reaction mixture was heated to room temperature under protection for 4-6 hours. After the reaction was complete, the mixture was extracted with dichloromethane and concentrated under vacuum to obtain product 1 (6.32 g, 16.03 mmol). Product 1 was dissolved in dry dichloromethane (160 mL, 0.1 M), the system was cooled to -20 °C, thiophenol (2.2 mL, 20.84 mmol) was added and stirred for 5 minutes, and then boron trifluoride ether solution (3.3 mL, 25.65 mmol) was added. 2The mixture was reacted at -20°C for 30 min under protection. After the reaction was complete, the pH was adjusted to 7-8 with triethylamine, extracted with dichloromethane, and concentrated under vacuum. The crude product was separated and purified by chromatography on a silica gel column (6:1, PE:EA) to obtain compound 3 (4.28 g, 63%).

[0106] S3: Remove the acetyl group of compound 3 to obtain compound 4. The reaction equation is as follows:

[0107]

[0108] The specific steps are as follows: sodium methoxide is dissolved in methanol to prepare a 0.3M sodium methoxide / methanol solution, compound 3 (4.28 g, 10.10 mmol) is mixed with 0.3M sodium methoxide / methanol solution (34 mL, 0.3M) and stirred for 30 min, and after the reaction is complete, the mixture is concentrated under reduced pressure in vacuo, and the crude product is separated and purified by chromatography on a silica gel column (20:1, DCM:MeOH) to obtain compound 4 (2.37 g, 79%);

[0109] S4: The 4- and 6-hydroxyl groups of compound 4 are protected with acetal, and the 3-hydroxyl group thereof is replaced with an -OTBS group to obtain compound 5. The reaction equation is as follows:

[0110]

[0111] The specific steps are as follows: Compound 4 (2.37 g, 7.98 mmol) was dissolved in dry acetonitrile (80 mL, 0.1 M), benzaldehyde dimethyl acetal (1.6 mL, 10.37 mmol) was added, and p-toluenesulfonic acid monohydrate (91 mg, 0.48 mmol) was added. 2 The mixture was reacted at room temperature for 3 h under protection. After the reaction was complete, triethylamine was added to adjust the pH to 7-8, and the mixture was extracted with dichloromethane and concentrated under vacuum to obtain intermediate 1 (2.16 g, 5.60 mmol). Intermediate 1 was dissolved in dry dichloromethane (28 mL, 0.2 M), activated imidazole (800 mg, 11.76 mmol) was added, and tert-butyldimethylsilyl chloride (1.77 g, 11.76 mmol) was added, and N 2 The reaction was allowed to proceed overnight at room temperature under protection. After the reaction was complete, the mixture was extracted with dichloromethane and concentrated under vacuum. The crude product was separated and purified by chromatography on a silica gel column (50:1, PE:EA) to obtain compound 5 (2.24 g, 80%).

[0112] S5: The acetal protecting group of compound 5 is removed, and the 4- and 6-hydroxyl groups are replaced with -OBz ​​groups to obtain compound 6. The reaction equation is as follows:

[0113]

[0114] The specific steps are as follows: acetic acid (23 mL, 403 mmol) and water (5.8 mL) are prepared into an 80% acetic acid aqueous solution, compound 5 (2.24 g, 4.48 mmol) is mixed therewith, N 2 The reaction mixture was stirred at 80°C for 20 min under protection. After the reaction was complete, the mixture was concentrated under vacuum to an oily substance. The crude product was separated and purified by chromatography on a silica gel column (8:1, PE:EA) to obtain product II (1.49 g, 3.63 mmol). Product II was dissolved in anhydrous pyridine (9 mL, 0.4 M), cooled to 0°C, benzoyl chloride (2.9 mL, 25.41 mmol) was added dropwise, the temperature was raised to room temperature, and N 2 The reaction was carried out under protection for 2 h. After the reaction was complete, methanol was added to quench the reaction, and the mixture was extracted with dichloromethane. The mixture was washed with 1N dilute hydrochloric acid, saturated sodium bicarbonate solution, and saturated brine. After vacuum concentration, the crude product was separated and purified by chromatography on a silica gel column (30:1, PE:EA) to obtain compound 6 (2.09 g, 93%).

[0115] S6: The amino group and carboxyl group of L-serine were protected with -Cbz and -Bn, respectively, to obtain compound 7. The reaction equation is as follows:

[0116]

[0117] The specific steps are as follows: sodium bicarbonate (2 g, 23.8 mmol) was dissolved in water (13 mL) and precooled at 0°C, L-serine (1 g, 9.52 mmol) was added in batches, stirred for 5 min, benzyl chloroformate (1.4 mL, 9.52 mmol) and toluene (1.4 mL) were prepared into a 50% solution, and the solution was slowly added dropwise into the system, and N 2 The mixture was stirred at 0°C for 2 h under protection, then heated to room temperature and stirred for 2 h. After the reaction was complete, ethyl acetate (50 mL) was added, and the pH was adjusted to 2.0 with 1N dilute hydrochloric acid. The mixture was extracted with ethyl acetate and concentrated under vacuum until one third of the liquid remained. Cyclohexane (100 mL) was added and stirred for 6 h. The mixture was filtered and dried to obtain the product III (1.13 g, 4.72 mmol). The product III was dissolved in N,N-dimethylformamide (47 mL, 0.1 M), and benzyl bromide (0.7 mL, 5.66 mmol), cesium carbonate (1.85 g, 5.66 mmol), N 2 The reaction was allowed to proceed overnight at room temperature under protection. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with dichloromethane and concentrated under reduced pressure in vacuo. The crude product was separated and purified by chromatography on a silica gel column (2:1, PE:EA) to obtain compound 7 (1.26 g, 81%).

[0118] S7: Compound 6 and compound 7 are subjected to glycosylation reaction to obtain compound 8. The reaction equation is as follows:

[0119]

[0120] The specific steps are as follows: Compound 6 (1.5 g, 2.42 mmol) was dissolved in dichloromethane (45 mL, 0.04 M), and then compound 7 (590 mg, 1.79 mmol) was added. The molecular sieves were stirred at 0°C for 10 min, and then 2 N-iodosuccinimide (545 mg, 2.42 mmol) was added under an aqueous solution of 4% paraformaldehyde (MW 400), and the mixture was stirred at 0°C for 5 min. Then trifluoromethanesulfonic acid (8 μL, 0.09 mmol) was slowly added. After the addition, the temperature was raised to room temperature and the reaction was continued for 2 h. After the reaction was completed, triethylamine was added to quench the reaction. The mixture was extracted with dichloromethane and concentrated under vacuum. The crude product was separated and purified by chromatography on a silica gel column (20:1, PE:EA) to obtain compound 8 (1.14 g, 76%).

[0121] 1 H NMR (600 MHz, CDCl 3 )δ8.04–8.02(m,2H),8.01–7.99(m,2H),7.59(t,J=7.2Hz,1H),7.53(t,J= 7.2Hz,1H),7.46(t,J=7.8Hz,2H),7.40(d,J=7.8Hz,2H),7.36(s,5H),7.31 (dd,J=6.3,3.9Hz,5H),5.83(d,J=8.4Hz,1H),5.56(d,J=3.6Hz,1H),5.22( d,J=2.4Hz,2H),5.05(s,2H),4.97(d,J=3.6Hz,1H),4.63(d,J=8.4Hz,1H), 4.43–4.38 (m, 1H), 4.30–4.26 (m, 2H), 4.13 (dd, J = 10.2, 3.3 Hz, 1H), 4.09 (dd, J = 10.8, 3.3 Hz, 1H), 4.03 (dd, J = 10.8, 3.0 Hz, 1H), 3.64 (dd, J = 10.2, 3.6 Hz, 1H), 0.77 (s, 9H), 0.17 (d, J = 6.6 Hz, 6H). δ4.97 (d, J = 3.6 Hz, 1H) is the hydrogen of the anomeric carbon of the sugar ring, and the coupling constant is 3.6, which belongs to the α-H category. When the reaction is at the hundred milligram level, it is α-only; when the reaction is at the nearly 2 gram level, the ratio of its peak area to the isomer is greater than 20:1.

[0122] 13 C NMR (151 MHz, CDCl 3)δ169.72,166.26,165.78,156.01,136.19,135.20,133.44,133.28,129.94,129.87,129.72,128.80,128.73,128.69,128.63,128 .56,128.41,128.30,99.26,70.94,69.60,68.26,67.88,67.31,63.37,61.87,54.63,25.65,17.86,-4.44,-5.00.HRMS(ESI)calcd for C 44 H 50 N 4 NaO 11 Si[M+Na] + 861.3143, found 861.3146.

[0123] S8: The -TBS group of compound 8 is removed to obtain compound 9. The reaction equation is as follows:

[0124]

[0125] The specific steps are as follows: Compound 8 (1.14 g, 1.36 mmol) was dissolved in dry dichloromethane (4.5 mL, 0.3 M), cooled to 0°C, and then a boron trifluoride ether solution (343 μL, 2.72 mmol) was added dropwise, and the mixture was heated to room temperature and stirred. The mixture was reacted overnight, and triethylamine was added to quench the reaction. The mixture was concentrated under reduced pressure in vacuo. The crude product was separated and purified by chromatography on a silica gel column (3:1, PE:EA) to obtain Compound 9 (788 mg, 80%).

[0126] S9: Compound 9 is subjected to a glycosylation reaction with 2,3,4,6-tetra-O-acetyl-α / β-D-galactopyranose trichloroacetimidate to obtain compound 10. The reaction equation is as follows:

[0127]

[0128] The specific steps are as follows: Compound 9 (400 mg, 0.55 mmol) was dissolved in dichloromethane (14 mL), and then compound 2,3,4,6-tetra-O-acetyl-α / β-D-pyranose trichloroacetimidate (380 mg, 0.77 mmol) was added, and then The molecular sieves were stirred at 0°C for 10 min, and then 2Trimethylsilyl trifluoromethanesulfonate (5 μL, 0.03 mmol) was slowly added under an atmosphere, and the mixture was kept warm (0°C) for 2 h. After the reaction was completed, triethylamine was added to quench the reaction. After vacuum concentration, the crude product was separated and purified by chromatography on a silica gel column (2:1, PE:EA) to obtain compound 10 (465 mg, 80%).

[0129] 1 H NMR (600 MHz, CDCl 3 )δ8.02(d,J=7.2Hz,2H),7.98(d,J=6.9Hz,2H),7.58(t,J=7.5Hz,1H),7.54–7.51(m,1H),7. 48–7.45(m,2H),7.38(dd,J=10.8,6.3Hz,7H),7.34–7.29(m,5H),5.77(d,J=8.4Hz,1H),5.7 4(d,J=3.6Hz,1H),5.29(d,J=3.9Hz,1H),5.22(d,J=2.7Hz,2H),5.12–5.06(m,3H),4.97(dd ,J=10.2,3.3Hz,1H),4.93(d,J=3.6Hz,1H),4.72(d,J=7.8Hz,1H),4.65–4.60(m,1H),4.40( dd,J=11.7,4.2Hz,1H),4.33(dd,J=11.7,7.8Hz,1H),4.25(dd,J=8.7,4.2Hz,1H),4.15(dd, J=10.8,3.6Hz,1H),4.10(dd,J=11.4,6.3Hz,1H),4.05(dd,J=10.5,3.3Hz,1H),4.02–3.96( m, 2H), 3.90 (t, J = 6.6 Hz, 1H), 3.72 (dd, J = 10.8, 3.6 Hz, 1H), 2.03 (s, 3H), 2.00 (s, 3H), 1.94 (s, 3H), 1.91 (s, 3H). δ4.72 (d, J = 7.8 Hz, 1H) is the hydrogen of the anomeric carbon of the galactose ring, and the coupling constant is 7.8, which belongs to the category of β-H, so it is determined that the main product is β configuration.

[0130] 13 C NMR (151 MHz, CDCl 3)δ170.56,170.31,170.24,169.69,169.52,165.69,155.98,136.11,135.16,133.36,1 33.28,130.12,129.85,129.82,129.71,128.84,128.79,128.73,128.63,128.57,128. 55,128.49,128.34,101.62,99.08,70.98,70.93,70.33,69.36,68.89,68.30,67.91,6 7.40,66.82,63.51,61.24,59.44,54.51,20.83,20.77,20.67,20.51.HRMS(ESI)calcd for C 52 H 55 N 4 O 20 [M+H] + 1055.3410, found 1055.3422.

[0131] S10: Compound 9 is subjected to group treatment to obtain compound 11 (Tn antigen). The reaction equation is as follows:

[0132]

[0133] The specific steps are as follows: 2 Under a nitrogen atmosphere, compound 9 (388 mg, 0.535 mmol) was dissolved in pyridine (13.4 mL, 0.04 M), and then thioacetic acid (6.5 mL, 91 mmol) was added dropwise. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by thin layer chromatography, the mixture was extracted with ethyl acetate and concentrated to obtain intermediate 2. Intermediate 2 was dissolved in methanol (13.4 mL), and then water (1.3 mL), glacial acetic acid (140 μL), and 10% palladium carbon (860 mg, 0.8 mmol) were added in sequence. Hydrogen was introduced into the reaction system, and the reaction was allowed to react for 6 h at room temperature. After the reaction was completed as monitored by thin layer chromatography, the reaction solution was filtered, and the filtrate was concentrated to obtain a white solid. Under nitrogen protection, the above white solid was dissolved in methanol (4.3 mL), and then 1,4-dioxane (4.3 mL) and 1M sodium hydroxide aqueous solution (6.4 mL) were added in sequence. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by thin layer chromatography, the pH was adjusted to 7.0 with glacial acetic acid, and the reaction solution was concentrated and Sephadex was used. LH-20 gel column chromatography purification (1:1, H 2 O: MeOH), to obtain compound 11 (Tn antigen) (58 mg, 35%);

[0134] 1 H NMR (600 MHz, D2 O) δ4.92(d,J=3.9Hz,1H),4.21(dd,J=11.1,3.9Hz,1H),4.13(dd,J=11.1,3.0Hz,1H),3.83–3.73(m,2H),2.05(s,3H).

[0135] 13 C NMR (101MHz, D 2 O)δ174.4,171.3,97.5,71.1,68.1,67.3,66.3,60.9,54.2,49.3,21.5.HRMS(ESI)calcd for C 11 H 19 N 2 O 8 [MH] - 307.1147, found 307.1148.

[0136] S11: Compound 10 is subjected to group treatment to obtain compound 12 (TF antigen). The reaction equation is as follows:

[0137]

[0138] The specific steps are as follows: 2 Under a nitrogen atmosphere, compound 10 (400 mg, 0.379 mmol) was dissolved in pyridine (9.5 mL, 0.04 M), and then thioacetic acid (4.6 mL, 64.4 mmol) was added dropwise, and the reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by thin layer chromatography, the reaction was extracted with ethyl acetate and concentrated to obtain intermediate three; intermediate three was dissolved in methanol (9.5 mL), and then water (950 μL), glacial acetic acid (100 μL), and 10% palladium carbon (310 mg, 0.569 mmol) were added in sequence, and hydrogen was introduced into the reaction system. The reaction was allowed to react for 6 h at room temperature. After the reaction was completed as monitored by thin layer chromatography, the reaction solution was filtered and the filtrate was concentrated to obtain a white solid; under nitrogen protection, the above white solid was dissolved in methanol (3 mL), and then 1,4-dioxane (3 mL) and 1M sodium hydroxide aqueous solution (4.6 mL) were added in sequence. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by thin layer chromatography, the pH was adjusted to 7.0 with glacial acetic acid, and the reaction solution was concentrated and Sephadex was used. LH-20 gel column chromatography purification (1:1, H 2 O: MeOH), to obtain compound 12 (TF antigen) (55 mg, 31%);

[0139] 1 H NMR (600MHz, D 2O)δ4.92(d,J=3.3Hz,1H),4.47(d,J=7.8Hz,1H),4.35(dd,J=11.1,3.3Hz,1H),4.23(s,1H),4.17–4.02(m,2H) ,4.04–3.95(m,2H),3.96–3.87(m,2H),3.84–3.72(m,4H),3.67–3.60(m,2H),3.55–3.49(m,1H),2.02(s,3H).

[0140] 13 C NMR (101MHz, D 2 O)δ174.7,171.9,104.7,98.5,76.8,75.3,72.6,71.3,70.6,68.7,68.7,61.4,61.1,48.5,22.2.HRMS(ESI)calcd for C 17 H 29 N 2 O 13 [MH] - 469.1675, found 469.1678.

[0141] Example 3

[0142] A method for stereospecifically synthesizing Tn antigen and TF antigen comprises the following steps:

[0143] S1: Compound 2 is obtained by addition reaction at the 1,2-position of compound 1. The reaction equation is as follows:

[0144]

[0145] The specific steps are as follows: Compound 1 (12.0 g, 44.1 mmol) was dissolved in dry acetonitrile (147 mL, 0.3 M), the system was cooled to -15 °C, ammonium cerium nitrate (60.44 g, 110.25 mmol) was added and stirred for 5 min, trimethylsilane (11.6 mL, 88.2 mmol) was added, and N 2 The temperature was raised to 0°C under protection and the reaction was allowed to proceed overnight. After the reaction was complete, ice water was added to the system to quench until the solid was completely dissolved, extracted with ethyl acetate, and concentrated under reduced pressure in vacuo. The crude product was separated and purified by chromatography on a silica gel column (5:1, PE:EA) to obtain compound 2 (7.14 g, 43%).

[0146] S2: -ONO in compound 2 2 After the group is replaced by a halogen atom Br, it is glycosylated with thiophenol to obtain compound 3. The reaction equation is as follows:

[0147]

[0148] The specific steps are as follows: Compound 2 (7.14 g, 18.97 mmol) was dissolved in dry acetonitrile (63 mL, 0.3 M), reacted under ice bath for 2 h, and then lithium bromide (9.9 g, 113.82 mmol) was added. 2 The reaction mixture was heated to room temperature under protection for 4-6 hours. After the reaction was complete, the mixture was extracted with dichloromethane and concentrated under vacuum to obtain product 1 (6.87 g, 17.42 mmol). Product 1 was dissolved in dry dichloromethane (58 mL, 0.3 M), the system was cooled to -20 °C, thiophenol (4.3 mL, 41.73 mmol) was added and stirred for 5 minutes, and then boron trifluoride ether solution (5.5 mL, 43.63 mmol) was added. 2 The mixture was reacted at -20°C for 30 min under protection. After the reaction was complete, the pH was adjusted to 7-8 with triethylamine, extracted with dichloromethane, and concentrated under vacuum. The crude product was separated and purified by chromatography on a silica gel column (6:1, PE:EA) to obtain compound 3 (4.26 g, 53%).

[0149] S3: Remove the acetyl group of compound 3 to obtain compound 4. The reaction equation is as follows:

[0150]

[0151] The specific steps are as follows: sodium methoxide is dissolved in methanol to prepare a 0.3M sodium methoxide / methanol solution, compound 3 (4.26 g, 10.06 mmol) is mixed with 0.3M sodium methoxide / methanol solution (20 mL, 0.5 M) and stirred for 30 min, and after the reaction is complete, the mixture is concentrated under vacuum, and the crude product is separated and purified by chromatography on a silica gel column (20:1, DCM:MeOH) to obtain compound 4 (2.48 g, 83%);

[0152] S4: The 4- and 6-hydroxyl groups of compound 4 are protected with acetal, and the 3-hydroxyl group thereof is replaced with an -OTBS group to obtain compound 5. The reaction equation is as follows:

[0153]

[0154] The specific steps are as follows: Compound 4 (2.48 g, 8.34 mmol) was dissolved in dry acetonitrile (28 mL, 0.3 M), benzaldehyde dimethyl acetal (2 mL, 13.34 mmol) was added, and p-toluenesulfonic acid monohydrate (158 mg, 0.83 mmol) was added. 2The mixture was reacted at room temperature for 3 h under protection. After the reaction was complete, triethylamine was added to adjust the pH to 7-8, and the mixture was extracted with dichloromethane and concentrated under vacuum to obtain intermediate 1 (2.15 g, 5.58 mmol). Intermediate 1 was dissolved in dry dichloromethane (11 mL, 0.5 M), activated imidazole (1 g, 15.06 mmol) was added, and tert-butyldimethylsilyl chloride (2.27 g, 15.06 mmol) was added. 2 The reaction was allowed to proceed overnight at room temperature under protection. After the reaction was complete, the mixture was extracted with dichloromethane and concentrated under vacuum. The crude product was separated and purified by chromatography on a silica gel column (50:1, PE:EA) to obtain compound 5 (2.29 g, 82%).

[0155] S5: The acetal protecting group of compound 5 is removed, and the 4- and 6-hydroxyl groups are replaced with -OBz ​​groups to obtain compound 6. The reaction equation is as follows:

[0156]

[0157] The specific steps are as follows: acetic acid (28.8 mL, 503.8 mmol) and water (7.2 mL) are prepared into an 80% acetic acid aqueous solution, compound 5 (2.29 g, 4.58 mmol) is mixed therewith, N 2 The reaction mixture was stirred at 80°C for 20 min under protection. After the reaction was complete, the mixture was concentrated under vacuum to an oily substance. The crude product was separated and purified by chromatography on a silica gel column (8:1, PE:EA) to obtain product II (1.49 g, 3.63 mmol). Product II was dissolved in anhydrous pyridine (6 mL, 0.6 M), cooled to 0°C, benzoyl chloride (3.8 mL, 32.67 mmol) was added dropwise, the temperature was raised to room temperature, and N 2 The reaction was carried out under protection for 2 h. After the reaction was complete, methanol was added to quench the reaction, and the mixture was extracted with dichloromethane. The mixture was washed with 1N dilute hydrochloric acid, saturated sodium bicarbonate solution, and saturated brine. After vacuum concentration, the crude product was separated and purified by chromatography on a silica gel column (30:1, PE:EA) to obtain compound 6 (2.16 g, 96%).

[0158] S6: The amino group and carboxyl group of L-serine were protected with -Cbz and -Bn, respectively, to obtain compound 7. The reaction equation is as follows:

[0159]

[0160] The specific steps are as follows: sodium bicarbonate (2.8 g, 33.32 mmol) was dissolved in water (16.6 mL) and precooled at 0°C, L-serine (1 g, 9.52 mmol) was added in batches, stirred for 5 min, benzyl chloroformate (2 mL, 14.28 mmol) and toluene (2 mL) were prepared into a 50% solution, and the solution was slowly added dropwise into the system, N 2The mixture was stirred at 0°C for 2 h under protection, then heated to room temperature and stirred for 2 h. After the reaction was complete, ethyl acetate (50 mL) was added, and the pH was adjusted to 2.0 with 1N dilute hydrochloric acid. The mixture was extracted with ethyl acetate and concentrated under vacuum until one third of the liquid remained. Cyclohexane (100 mL) was added and stirred for 6 h. The mixture was filtered and dried to obtain the product III (1.16 g, 4.86 mmol). The product III was dissolved in N,N-dimethylformamide (16.2 mL, 0.3 M), and benzyl bromide (0.9 mL, 7.29 mmol), cesium carbonate (2.37 g, 7.29 mmol), N 2 The reaction was allowed to proceed overnight at room temperature under protection. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with dichloromethane and concentrated under vacuum. The crude product was separated and purified by chromatography on a silica gel column (2:1, PE:EA) to obtain compound 7 (1.15 g, 72%).

[0161] S7: Compound 6 and compound 7 are subjected to glycosylation reaction to obtain compound 8. The reaction equation is as follows:

[0162]

[0163] The specific steps are as follows: Compound 6 (2.16 g, 3.48 mmol) was dissolved in dichloromethane (39 mL, 0.06 M), and then compound 7 (765 mg, 2.32 mmol) was added. The molecular sieves were stirred at 0°C for 10 min, and then 2 N-iodosuccinimide (785 mg, 3.48 mmol) was added under an aqueous solution of 4,4-dichloromethane (2,4-dichloroethane) and stirred at 0°C for 5 min. Then trifluoromethanesulfonic acid (30 μL, 0.348 mmol) was slowly added. After the addition, the temperature was raised to room temperature and the reaction was continued for 2 h. After the reaction was completed, triethylamine was added to quench the reaction. The mixture was extracted with dichloromethane and concentrated under vacuum. The crude product was separated and purified by chromatography on a silica gel column (20:1, PE:EA) to obtain compound 8 (1.62 g, 83%).

[0164] 1 H NMR (600 MHz, CDCl 3)δ8.04–8.02(m,2H),8.01–7.99(m,2H),7.59(t,J=7.2Hz,1H),7.53(t,J= 7.2Hz,1H),7.46(t,J=7.8Hz,2H),7.40(d,J=7.8Hz,2H),7.36(s,5H),7.31 (dd,J=6.3,3.9Hz,5H),5.83(d,J=8.4Hz,1H),5.56(d,J=3.6Hz,1H),5.22( d,J=2.4Hz,2H),5.05(s,2H),4.97(d,J=3.6Hz,1H),4.63(d,J=8.4Hz,1H), 4.43–4.38 (m, 1H), 4.30–4.26 (m, 2H), 4.13 (dd, J = 10.2, 3.3 Hz, 1H), 4.09 (dd, J = 10.8, 3.3 Hz, 1H), 4.03 (dd, J = 10.8, 3.0 Hz, 1H), 3.64 (dd, J = 10.2, 3.6 Hz, 1H), 0.77 (s, 9H), 0.17 (d, J = 6.6 Hz, 6H). δ4.97 (d, J = 3.6 Hz, 1H) is the hydrogen of the anomeric carbon of the sugar ring, and the coupling constant is 3.6, which belongs to the α-H category. When the reaction is at the hundred milligram level, it is α-only; when the reaction is at the nearly 2 gram level, the ratio of its peak area to the isomer is greater than 20:1.

[0165] 13 C NMR (151 MHz, CDCl 3 )δ169.72,166.26,165.78,156.01,136.19,135.20,133.44,133.28,129.94,129.87,129.72,128.80,128.73,128.69,128.63,128 .56,128.41,128.30,99.26,70.94,69.60,68.26,67.88,67.31,63.37,61.87,54.63,25.65,17.86,-4.44,-5.00.HRMS(ESI)calcd for C 44 H 50 N 4 NaO 11 Si[M+Na] + 861.3143, found 861.3146.

[0166] S8: The -TBS group of compound 8 is removed to obtain compound 9. The reaction equation is as follows:

[0167]

[0168] The specific steps are as follows: Compound 8 (1.62 g, 1.93 mmol) was dissolved in dry dichloromethane (4 mL, 0.5 M), cooled to 0°C, and then a boron trifluoride ether solution (975 μL, 7.72 mmol) was added dropwise, and the mixture was heated to room temperature and stirred. The mixture was reacted overnight, and triethylamine was added to quench the reaction. The mixture was concentrated under reduced pressure in vacuo. The crude product was separated and purified by chromatography on a silica gel column (3:1, PE:EA) to obtain Compound 9 (980 mg, 70%).

[0169] S9: Compound 9 is subjected to a glycosylation reaction with 2,3,4,6-tetra-O-acetyl-α / β-D-galactopyranose trichloroacetimidate to obtain compound 10. The reaction equation is as follows:

[0170]

[0171] The specific steps are as follows: Compound 9 (500 mg, 0.69 mmol) was dissolved in dichloromethane (11.5 mL, 0.06 M), and then compound 2,3,4,6-tetra-O-acetyl-α / β-D-pyranose trichloroacetimidate (545 mg, 1.1 mmol) was added. The molecular sieves were stirred at 0°C for 10 min, and then 2 Trimethylsilyl trifluoromethanesulfonate (18 μL, 0.1 mmol) was slowly added under an atmosphere, and the mixture was kept warm (0°C) for 2 h. After the reaction was completed, triethylamine was added to quench the reaction. After vacuum concentration, the crude product was separated and purified by chromatography on a silica gel column (2:1, PE:EA) to obtain compound 10 (582 mg, 80%).

[0172] 1 H NMR (600 MHz, CDCl 3)δ8.02(d,J=7.2Hz,2H),7.98(d,J=6.9Hz,2H),7.58(t,J=7.5Hz,1H),7.54–7.51(m,1H),7. 48–7.45(m,2H),7.38(dd,J=10.8,6.3Hz,7H),7.34–7.29(m,5H),5.77(d,J=8.4Hz,1H),5.7 4(d,J=3.6Hz,1H),5.29(d,J=3.9Hz,1H),5.22(d,J=2.7Hz,2H),5.12–5.06(m,3H),4.97(dd ,J=10.2,3.3Hz,1H),4.93(d,J=3.6Hz,1H),4.72(d,J=7.8Hz,1H),4.65–4.60(m,1H),4.40( dd,J=11.7,4.2Hz,1H),4.33(dd,J=11.7,7.8Hz,1H),4.25(dd,J=8.7,4.2Hz,1H),4.15(dd, J=10.8,3.6Hz,1H),4.10(dd,J=11.4,6.3Hz,1H),4.05(dd,J=10.5,3.3Hz,1H),4.02–3.96( m, 2H), 3.90 (t, J = 6.6 Hz, 1H), 3.72 (dd, J = 10.8, 3.6 Hz, 1H), 2.03 (s, 3H), 2.00 (s, 3H), 1.94 (s, 3H), 1.91 (s, 3H). δ4.72 (d, J = 7.8 Hz, 1H) is the hydrogen of the anomeric carbon of the galactose ring, and the coupling constant is 7.8, which belongs to the category of β-H, so it is determined that the main product is β configuration.

[0173] 13 C NMR (151 MHz, CDCl 3 )δ170.56,170.31,170.24,169.69,169.52,165.69,155.98,136.11,135.16,133.36,1 33.28,130.12,129.85,129.82,129.71,128.84,128.79,128.73,128.63,128.57,128. 55,128.49,128.34,101.62,99.08,70.98,70.93,70.33,69.36,68.89,68.30,67.91,6 7.40,66.82,63.51,61.24,59.44,54.51,20.83,20.77,20.67,20.51.HRMS(ESI)calcd for C 52 H55 N 4 O 20 [M+H] + 1055.3410, found 1055.3422.

[0174] S10: Compound 9 is subjected to group treatment to obtain compound 11 (Tn antigen). The reaction equation is as follows:

[0175]

[0176] The specific steps are as follows: 2 Under a nitrogen atmosphere, compound 9 (480 mg, 0.66 mmol) was dissolved in pyridine (11 mL, 0.06 M), and then thioacetic acid (8.9 mL, 125.4 mmol) was added dropwise. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by thin layer chromatography, the mixture was extracted with ethyl acetate and concentrated to obtain intermediate 2. Intermediate 2 was dissolved in methanol (30 mL), and then water (3 mL), glacial acetic acid (300 μL), and 10% palladium carbon (1.77 g, 1.65 mmol) were added in sequence. Hydrogen was introduced into the reaction system, and the reaction was allowed to react for 6 h at room temperature. After the reaction was completed as monitored by thin layer chromatography, the reaction solution was filtered, and the filtrate was concentrated to obtain a white solid. Under nitrogen protection, the above white solid was dissolved in methanol (7.9 mL), and then 1,4-dioxane (7.9 mL) and 1M sodium hydroxide aqueous solution (12 mL) were added in sequence. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by thin layer chromatography, the pH was adjusted to 7.0 with glacial acetic acid, and the reaction solution was concentrated and Sephadex was used. LH-20 gel column chromatography purification (1:1, H 2 O: MeOH), to obtain compound 11 (Tn antigen) (73 mg, 36%);

[0177] 1 H NMR (600MHz, D 2 O) δ4.92(d,J=3.9Hz,1H),4.21(dd,J=11.1,3.9Hz,1H),4.13(dd,J=11.1,3.0Hz,1H),3.83–3.73(m,2H),2.05(s,3H).

[0178] 13 C NMR (101MHz, D 2 O)δ174.4,171.3,97.5,71.1,68.1,67.3,66.3,60.9,54.2,49.3,21.5.HRMS(ESI)calcd for C 11 H 19 N 2 O 8 [MH]- 307.1147, found 307.1148.

[0179] S11: Compound 10 is subjected to group treatment to obtain compound 12 (TF antigen). The reaction equation is as follows:

[0180]

[0181] The specific steps are as follows: 2 Under atmosphere, compound 10 (582 mg, 0.55 mmol) was dissolved in pyridine (9.2 mL, 0.06 M), and then thioacetic acid (7.4 mL, 104.5 mmol) was added dropwise. The reaction was allowed to react overnight at room temperature. After the reaction was completed as monitored by thin layer chromatography, the mixture was extracted with ethyl acetate and concentrated to obtain intermediate 3. Intermediate 3 was dissolved in methanol (25 mL), and then water (2.5 mL), glacial acetic acid (250 μL), 10% palladium carbon (1.47 g, 1.37 5mmol), hydrogen was introduced into the reaction system, and the reaction was carried out at room temperature for 6h. After the reaction was completed as monitored by thin layer chromatography, the reaction solution was filtered and the filtrate was concentrated to obtain a white solid. Under nitrogen protection, the white solid was dissolved in methanol (6.6mL), and 1,4-dioxane (6.6mL) and 1M sodium hydroxide aqueous solution (10mL) were added in sequence, and the reaction was carried out at room temperature overnight. After the reaction was completed as monitored by thin layer chromatography, glacial acetic acid was used to adjust the pH to 7.0, and the reaction solution was concentrated and purified by Sephadex LH-20 gel column chromatography (1:1, H 2 O: MeOH), to obtain compound 12 (TF antigen) (80 mg, 31%);

[0182] 1 H NMR (600MHz, D 2 O)δ4.92(d,J=3.3Hz,1H),4.47(d,J=7.8Hz,1H),4.35(dd,J=11.1,3.3Hz,1H),4.23(s,1H),4.17–4.02(m,2H) ,4.04–3.95(m,2H),3.96–3.87(m,2H),3.84–3.72(m,4H),3.67–3.60(m,2H),3.55–3.49(m,1H),2.02(s,3H).

[0183] 13 C NMR (101MHz, D 2 O)δ174.7,171.9,104.7,98.5,76.8,75.3,72.6,71.3,70.6,68.7,68.7,61.4,61.1,48.5,22.2.HRMS(ESI)calcd for C17 H 29 N 2 O 13 [M-H] - 469.1675,found469.1678.

Claims

1. A method for stereospecifically synthesizing Tn antigen and TF antigen, characterized in that: The following steps are involved: S1: Using 3,4,6-tri-O-acetyl-galactenose as the initial substrate, compound 1 is subjected to an addition reaction at the 1,2-position of compound 1 to obtain compound 2. S2: Replace the -ONO2 group in compound 2 with Br, and then react with thiophenol to obtain compound 3. S3: remove all acetyl groups of compound 3 to obtain compound 4, S4: Protect the 4- and 6-hydroxyl groups of compound 4 with acetal, and replace the 3-hydroxyl group with -OTBS group to obtain compound 5. S5: removing the acetal protecting group of compound 5 and replacing the 4,6-hydroxyl groups with -OBz ​​groups to obtain compound 6. S6: Protect the amino group and carboxyl group of L-serine with -Cbz and -Bn respectively to obtain compound 7. S7: Compound 6 and compound 7 are subjected to glycosylation reaction to obtain compound 8. S8: removing the -TBS group of compound 8 to obtain compound 9, S9: Compound 9 is subjected to a glycosylation reaction with 2,3,4,6-tetra-O-acetyl-α / β-D-galactopyranose trichloroacetimidate to obtain compound 10. S10: Compound 9 and compound 10 are subjected to group treatment to obtain compound 11 and compound 12.

2. The method for stereospecific synthesis of Tn antigen and TF antigen according to claim 1, characterized in that: The specific preparation process is as follows: S1: Compound 1 was dissolved in dry acetonitrile, the system was placed below 0°C with stirring and precooled, ammonium cerium nitrate was added, and trimethylsilyl azide was added dropwise, the reaction was carried out in an ice bath under N2 protection overnight, ice water was added until the solid was dissolved, and the mixture was extracted with ethyl acetate, dried, concentrated, and purified to obtain compound 2; S2: Compound 2 is dissolved in dry acetonitrile, precooled at 0°C, and lithium bromide is added. The mixture is heated to room temperature and reacted for 4-6 hours. The mixture is extracted with dichloromethane, dried, and concentrated to obtain product 1. Product 1 is dissolved in dry dichloromethane, and the mixture is placed in an environment below -20°C, thiophenol is added, stirred, and mixed. Boron trifluoride ether solution is added dropwise, and the mixture is reacted under N2 protection. Triethylamine is added to adjust the pH to 7-8. The mixture is extracted with dichloromethane, dried, concentrated, and purified to obtain compound 3. S3: Compound 3 was mixed with MeONa / MeOH solution, stirred at room temperature for sufficient reaction, concentrated and purified to obtain compound 4; S4: Compound 4 is dissolved in dry acetonitrile, and then benzaldehyde dimethyl acetal and p-toluenesulfonic acid are added in sequence, and the mixture is allowed to react fully at room temperature, and triethylamine is added to adjust the pH to 7-8, and the mixture is concentrated, extracted with dichloromethane, dried, and concentrated to obtain intermediate 1; intermediate 1 is dissolved in dry dichloromethane, stirred at room temperature, and activated imidazole and tert-butyldimethylsilyl chloride are added in sequence, and the mixture is allowed to react overnight at room temperature, extracted with dichloromethane, dried, concentrated, and purified to obtain compound 5; S5: Compound 5 is mixed with acetic acid aqueous solution and stirred, heated under N2 protection, condensed and refluxed, concentrated, and purified to obtain product 2; product 2 is dissolved in anhydrous pyridine, benzoyl chloride is slowly added dropwise at 0°C, the temperature is raised to 35-45°C for sufficient reaction, methanol is added for quenching, dichloromethane is extracted, HCl is added for washing until the water layer becomes acidic, dried, concentrated, and purified to obtain compound 6; S6: Add L-serine in batches to a 0°C precooled sodium bicarbonate aqueous solution, cool to 0°C, slowly drop a mixed solution of benzyl chloroformate and toluene, continue stirring at 0°C for 1-3h, warm to room temperature and stir for 1-3h, add ethyl acetate, adjust the pH to 2.0 with dilute hydrochloric acid, extract with ethyl acetate, concentrate to obtain a suspension, add cyclohexane and stir to react fully, filter and dry to obtain product 3; under nitrogen protection, dissolve product 3 in dry N,N-dimethylformamide, add benzyl bromide and cesium carbonate in sequence, stir at room temperature overnight, quench with water, extract with dichloromethane, concentrate and purify to obtain compound 7; S7: Compound 6 and Compound 7 were dissolved in dry dichloromethane and added molecular sieves, mix thoroughly below 0°C, then add N-iodosuccinimide in an inert gas atmosphere, stir and mix at 0°C, then add trifluoromethanesulfonic acid, warm to room temperature and react for 1-3 hours, then add triethylamine to adjust the pH to 7-8, concentrate, and purify to obtain compound 8; S8: Compound 8 was dissolved in dry dichloromethane, and a boron trifluoride ether solution was added dropwise under an ice bath, and the mixture was heated to room temperature and stirred to react overnight, and triethylamine was added to adjust the pH to 7-8, and the mixture was concentrated and purified to obtain compound 9; S9: Compound 9 and 2,3,4,6-tetra-O-acetyl-α / β-D-galactopyranose trichloroacetimidate were dissolved in dry dichloromethane and added molecular sieves, stirring and mixing at 0°C, then adding trimethylsilyl trifluoromethanesulfonate in an inert gas atmosphere, continuing the reaction at 0°C for 1-3 hours, then adding triethylamine to adjust the pH to 7-8, concentrating, and purifying to obtain compound 10; S10: Under nitrogen protection, compound 9 was dissolved in pyridine, and then thioacetic acid was added dropwise. After stirring at room temperature for overnight reaction, the mixture was concentrated and purified to obtain intermediate 2. Intermediate 2 was dissolved in methanol, and water, glacial acetic acid, and 10% palladium carbon were added in sequence. Hydrogen was introduced into the reaction system. After the reaction was complete at room temperature, the mixture was filtered and concentrated to obtain a white solid. Under nitrogen protection, the white solid was dissolved in methanol, and 1,4-dioxane and sodium hydroxide aqueous solution were added in sequence. The mixture was reacted at room temperature overnight. The pH was adjusted to neutral with glacial acetic acid. The reaction solution was concentrated and purified to obtain compound 11. Compound 10 was subjected to the above-mentioned operation steps to obtain compound 12.

3. The method for stereospecific synthesis of Tn antigen and TF antigen according to claim 2, characterized in that: In S1, the concentration of compound 1 in acetonitrile is 0.1-0.3 mmol / mL, and the molar ratio of compound 1, ammonium cerium nitrate and trimethylsilyl azide is 0.8-1.2:1.5-2.5:1.5-2; In S2, the concentration of compound 2 in acetonitrile is 0.1-0.3 mmol / mL, and the molar ratio of compound 2 to lithium bromide is 1:4-6; the concentration of product 1 in dichloromethane is 0.1-0.3 mmol / mL, and the molar ratio of product 1, thiophenol and boron trifluoride ether is 1:1.3-2.2:1.3-2.

3.

4. The method for stereospecific synthesis of Tn antigen and TF antigen according to claim 2, characterized in that: The concentration of compound 3 in MeONa / MeOH in S3 is 0.3-0.5 mmol / mL; the reaction time is 20-40 min; In S4, the concentration of compound 4 in acetonitrile is 0.1-0.3 mmol / mL, and the molar ratio of compound 4, benzaldehyde dimethyl acetal and p-toluenesulfonic acid is 1:1.3-1.6:0.06-0.1; the concentration of intermediate 1 in dichloromethane is 0.2-0.5 mmol / mL, and the molar ratio of intermediate 1, imidazole and tert-butyldimethylchlorosilane is 1:2.1-2.7:2.1-2.

7.

5. The method for stereospecific synthesis of Tn antigen and TF antigen according to claim 2, characterized in that: The molar ratio of compound 5 to acetic acid in S5 is 1:90-110, the concentration of product II in pyridine is 0.4-0.6 mmol / mL, and the molar ratio of product II to benzoyl chloride is 1:7-9; The concentration of the sodium bicarbonate aqueous solution in S6 is 1.8-2.0 mmol / mL, the molar ratio of L-serine, sodium bicarbonate and benzyl chloroformate is 1:2.5-3.5:1-1.5, the concentration of product three in N,N-dimethylformamide is 0.1-0.3 mmol / mL, and the molar ratio of product three, benzyl bromide and cesium carbonate is 1:1-1.5:1-1.

5.

6. The method for stereospecific synthesis of Tn antigen and TF antigen according to claim 2, characterized in that: The concentration of compound 7 in dichloromethane in S7 is 0.04-0.06 mmol / mL, and the molar ratio of compound 6, compound 7, N-iodosuccinimide and trifluoromethanesulfonic acid is 1.2-1.5:1:1-1.4:0.05-0.

15.

7. The method for stereospecific synthesis of Tn antigen and TF antigen according to claim 2, characterized in that: In S8, the concentration of compound 8 in dichloromethane is 0.3-0.5 mmol / mL, and the molar ratio of compound 8 to boron trifluoride ether is 1:2-4; The concentration of compound 9 in dichloromethane in S9 is 0.04-0.06 mmol / mL, and the molar ratio of compound 9, 2,3,4,6-tetra-O-acetyl-α / β-D-pyranose trichloroacetimidate and trimethylsilyl trifluoromethanesulfonate is 1:1.4-1.6:0.05-0.

15.

8. The method for stereospecific synthesis of Tn antigen and TF antigen according to claim 2, characterized in that: The concentration of compound 9 in S10 in pyridine solution is 0.04-0.06 mmol / mL, and the molar ratio of compound 9 to thioacetic acid is 1:170-190; the molar volume ratio of intermediate 2, methanol, water, and glacial acetic acid is 1 mmol:25-45 mL:2.5-4.5 mL:250-450 μL, and the molar ratio of intermediate 2 to palladium carbon is 1:1.5-2.5; the molar volume ratio of white solid, methanol, 1,4-dioxane, and 1M sodium hydroxide aqueous solution is 1 mmol:8-12 mL:8-12 mL:12-18 mL.

9. The method for stereospecific synthesis of Tn antigen and TF antigen according to claim 2, characterized in that: The concentration of compound 10 in S10 in pyridine solution is 0.04-0.06 mmol / mL, and the molar ratio of compound 10 to thioacetic acid is 1:170-190; the molar volume ratio of intermediate three, methanol, water, and glacial acetic acid is 1 mmol:25-45 mL:2.5-4.5 mL:250-450 μL, and the molar ratio of intermediate three to palladium carbon is 1:1.5-2.5; the molar volume ratio of white solid, methanol, 1,4-dioxane, and 1M sodium hydroxide aqueous solution is 1 mmol:8-12 mL:8-12 mL:12-18 mL.

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