A kind of black wolfberry oligosaccharide derivative and its preparation method and application

Abstract

The present invention belongs to the technical field of chemical synthesis, and in particular relates to a ruthenium wolfberry oligosaccharide derivative, a preparation method and application thereof. The invention utilizes the remote participation effect of acyl groups, steric hindrance effect, solvent effect and end group effect to achieve efficient construction of 1,2-cis-α-galacturonic acid glycosidic bonds and other glycosidic bonds in the oligosaccharide structure, thereby realizing the first chemical synthesis of ruthenium wolfberry oligosaccharide and its derivatives. The ruthenium wolfberry oligosaccharide fragment provided by the present invention has an inhibitory effect on pancreatic cancer cells, provides a theoretical basis for clarifying the active fragments (active structural domains) of ruthenium wolfberry polysaccharides, conducting more in-depth functional mechanism research and developing new anti-pancreatic cancer drugs based on the active fragments, and has good application prospects in the preparation of new anti-cancer drugs.

Description

Technical Field

[0001] The present invention belongs to the technical field of chemical synthesis, and in particular relates to a lycium barbarum oligosaccharide derivative and a preparation method and application thereof. Background Art

[0002] Pancreatic cancer is one of the most lethal malignancies. Compared to other malignancies, pancreatic cancer is highly aggressive, with an insidious onset and a lack of typical symptoms. It is also highly invasive, with the potential for tissue invasion and distant metastasis early on. Surgical resection rates are low, recurrence rates are high, and the prognosis is poor. The five-year survival rate is only 8%, and for patients with locally advanced or metastatic pancreatic cancer, the five-year survival rate is as low as 2%. Currently, treatment options for pancreatic cancer include surgery, embolization, radiotherapy, chemotherapy, and immunotherapy. Surgery is the only potentially curative option, but only 20% of patients are eligible. Even for these patients who undergo radical surgery, the risk of postoperative recurrence and metastasis remains high. Chemotherapy is one of the mainstays of pancreatic cancer treatment, using gemcitabine as the primary drug, sometimes in combination with other drugs such as 5-fluorouracil. However, patients can easily develop resistance to these drugs during treatment. Consequently, treatment options for pancreatic cancer are extremely limited, and the mortality rate remains high, necessitating the development of new treatment options and drugs.

[0003] Traditional Chinese medicine polysaccharides have significant anti-tumor activity and minimal toxic side effects, demonstrating their enormous potential for application. Studies have shown that naturally extracted polysaccharides, such as Lycium ruthenicum polysaccharides, Panax notoginseng flower polysaccharides, safflower polysaccharides, Polygala tenuifolia polysaccharides, corn polysaccharides, Ophiopogon japonicus polysaccharides, honeysuckle polysaccharides, and Gastrodia elata polysaccharides, exhibit significant anti-pancreatic cancer activity and are important lead compounds for the development of new anti-pancreatic cancer drugs. Lycium ruthenicum Murr is a perennial shrub of the Solanaceae family, containing a variety of active ingredients, including polysaccharides, flavonoids, anthocyanins, phenolic acids, and essential oils. Previous research by the research team isolated and isolated two homogeneous polysaccharides, LRP1-S2 and LRP3-S1, from Lycium ruthenicum (Lycium barbarum), with relative molecular masses of 17.0 kDa and 114.8 kDa, respectively. The structures of the two polysaccharides were also characterized. In vitro activity experiments demonstrated that both LRP1-S2 and LRP3-S1 inhibited the proliferation of pancreatic cancer cells BxPC-3, AsPC-1, and PANC-1, while exhibiting no significant cytotoxicity against normal pancreatic cells HPDE6-C7 and hepatocytes LO2. Furthermore, LRP1-S2 exhibited a significant inhibitory effect on solid tumor growth in a nude mouse model of pancreatic cancer BxPC-3 xenografts, with an inhibitory effect superior to that achieved by the gemcitabine positive control group. Further mechanistic studies showed that LRP1-S2 may induce apoptosis of BxPC-3 cells by blocking the p38 MAPK / NF-κB and GSK-3β / β-Catenin signaling pathways; LRP3-S1 can weaken the invasive ability of pancreatic cancer cells BxPC-3, which may be due to the fact that this polysaccharide blocks the p38 MAPK signaling pathway and the FAK / AKT / GSK-3β signaling pathway.

[0004] The two polysaccharides can block the same signaling pathway, which may be closely related to their structures. Lycium ruthenicum polysaccharide LRP1-S2 is an arabinogalactan, while LRP3-S1 is a typical type I rhamnogalacturonan (RG-I). These two polysaccharides have unique structures and are important lead compounds for the development of new anti-pancreatic cancer carbohydrate drugs. However, their active fragments (active determinants) are still unclear. Furthermore, the discovery of the anti-pancreatic cancer target molecules of the two polysaccharides and the related mechanistic research are not yet in-depth and systematic, and are still in their infancy. Obtaining structurally well-defined and uniform polysaccharides through natural extraction methods is extremely challenging. The quality of polysaccharides is affected by many factors, including climate, cultivation conditions, and extraction methods. Polysaccharides have a relatively large molecular weight and relatively poor solubility, which also affects their ability to enter the body and produce pharmacological activity. Purification efficiency is low, structure determination is difficult, and obtaining sufficient amounts of well-defined oligosaccharide fragments is difficult, limiting their application in carbohydrate drug development. Therefore, preparing structurally clear and uniform oligosaccharide fragments through chemical synthesis and exploring the active fragments (active determinants) of the two polysaccharides are crucial for the research and development and production of innovative anti-pancreatic cancer drugs based on Lycium barbarum polysaccharides. Summary of the Invention

[0005] The purpose of the present invention is to synthesize oligosaccharide fragments related to black wolfberry polysaccharides LRP1-S2 and LRP3-S1 based on a chemical method, explore the active fragments (active determinants) of the two polysaccharides, and provide a theoretical basis for the research and development and production of anti-pancreatic cancer carbohydrate drugs.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is:

[0007] A lycium barbarum oligosaccharide derivative, wherein the derivative is a compound having a structure represented by formula (1) to (8) or a pharmaceutically acceptable salt thereof:

[0008]

[0009] Among them, PG1, PG5, PG 10 PG 17 is any one of hydroxy, acetoxy, benzoyloxy, pivaloyloxy, chloroacetoxy, levulinyloxy, allyloxycarbonyloxy, methoxy, benzyloxy, 2-naphthylmethoxy, p-methoxybenzyloxy, allyloxy, p-methoxyphenoxy, trityloxy, monomethoxytrityloxy, dimethoxytrityloxy, tert-butyldimethylsilyloxy, tert-butyldiphenylsilyloxy, triethylsilyloxy, halogen, trichloroacetimidyloxy, N-phenyltrifluoroacetimidyloxy, dibenzylphosphoxy, o-alkynylbenzoyloxy, o-methoxycarbonylalkynylphenylthio, methylthio, ethylthio, phenylthio, and p-tolylthio; PG2, PG6, PG 11 PG 14 It can be any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, levulinyl, and allyloxycarbonyl; PG3, PG4, PG7, PG8, PG9, PG 12 PG 15 PG 16 PG is any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, levulinyl, allyloxycarbonyl, methyl, benzyl, 2-naphthylmethyl, p-methoxybenzyl, allyl, p-methoxyphenyl, trityl, monomethoxytrityl, dimethoxytrityl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, triethylsilyl, benzylideneacetal, and isopropylideneketal; 13 Any one of hydrogen, methyl, ethyl, tert-butyl, benzyl, allyl, trichloroethyl, and trimethylsilylethyl; PG 18 It is any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, levulinyl and allyloxycarbonyl.

[0010] Furthermore, the pharmaceutically acceptable salt is any one of sodium salt, potassium salt, calcium salt, magnesium salt, zinc salt and meglumine salt.

[0011] A method for preparing a lycium barbarum oligosaccharide derivative comprises the following steps:

[0012] S1. Glycosylation of glycosyl donor 9 and glycosyl acceptor 10 yields disaccharide 1. Acyl protecting groups of disaccharide 1 are removed under alkaline conditions, followed by reduction to remove aromatic protecting groups, completing deprotection and yielding fully deprotected disaccharide 1a. The synthetic route is as follows:

[0013]

[0014] wherein LG is a leaving group of the glycosyl donor and is any one of halogen, trichloroacetimidyloxy, N-phenyltrifluoroacetimidyloxy, dibenzylphosphoxy, o-alkynylbenzoyloxy, o-methoxycarbonylalkynylphenylthio, methylthio, ethylthio, phenylthio, and p-tolylthio;

[0015] S2. Disaccharide 1 is selectively deprotected by PG4 to yield disaccharide 11, which is then glycosylated with glycosyl donor 9 under the catalytic action of a promoter to yield trisaccharide 2. Trisaccharide 2 is then deprotected by removing the acyl protecting group under alkaline conditions and the aromatic protecting group by reduction, completing deprotection and yielding fully deprotected trisaccharide 2a. The synthetic route is shown below:

[0016]

[0017] S3. Glycosidation of glycosyl donor 9 and glycosyl acceptor 12 yields disaccharide 3. Acyl protecting groups of disaccharide 3 are removed under alkaline conditions, followed by reduction to remove aromatic protecting groups, completing deprotection and yielding fully deprotected trisaccharide 3a. The synthetic route is as follows:

[0018]

[0019] S4. Glycosylation of glycosyl donor 13 and glycosyl acceptor 14 yields disaccharide 4. Acyl protecting groups of disaccharide 4 are removed under alkaline conditions, followed by reduction to remove aromatic protecting groups, completing deprotection and yielding fully deprotected trisaccharide 4a. The synthetic route is as follows:

[0020]

[0021] S5. Glycosylation of glycosyl donor 13 and glycosyl acceptor 15 yields disaccharide 5. Acyl protecting groups of disaccharide 5 are removed under alkaline conditions, followed by reduction to remove aromatic protecting groups, completing deprotection and yielding fully deprotected trisaccharide 5a. The synthetic route is as follows:

[0022]

[0023] S6. Glycosylation of glycosyl donor 16 and glycosyl acceptor 14 yields disaccharide 6. The acyl protecting group of disaccharide 6 is removed under alkaline conditions, and the aromatic protecting group is removed by reduction to complete deprotection, yielding fully deprotected trisaccharide 6a. The synthetic route is as follows:

[0024]

[0025] S7. Glycosylation of glycosyl donor 17 and glycosyl acceptor 18 is performed, utilizing the remote participation effect of the C4 acyl group to complete the construction of a 1,2-cis-glycosidic bond, yielding disaccharide 7. Disaccharide 7 is then deprotected by removing the acyl protecting group under alkaline conditions, followed by reduction to remove the aromatic protecting group, yielding fully deprotected disaccharide 7a. The synthetic route is as follows:

[0026]

[0027] S8. Glycosylation of glycosyl donor 19 and glycosyl acceptor 14 yields trisaccharide 8. Trisaccharide 7 is deprotected by removing the acyl protecting group under alkaline conditions, followed by reduction to remove the aromatic protecting group, thereby completing deprotection and yielding fully deprotected trisaccharide 8a. The synthetic route is as follows:

[0028]

[0029] Furthermore, the accelerator is any one of boron trifluoride etherate, trifluoromethanesulfonic acid, trimethylsilyl trifluoromethanesulfonate, silver carbonate, trifluoromethanesulfonic acid, N-iodosuccinimide and trifluoromethanesulfonic acid, N-iodosuccinimide and N-iodosuccinimide and trifluoromethanesulfonic acid, triphenylphosphine gold trifluoromethanesulfonate, and triphenylphosphine gold bis(fluorosulfonyl)imide.

[0030] Furthermore, the reduction reaction conditions for removing the aromatic protecting group are: a mixed solvent consisting of dichloromethane or tetrahydrofuran or ethyl acetate and tert-butanol or methanol, water, and acetic acid, a 10% palladium carbon catalyst or palladium hydroxide, and hydrogen, and stirring at room temperature for 48 hours; or sodium, liquid ammonia, tetrahydrofuran, and tert-butanol, and stirring at -78°C for 30 minutes.

[0031] Furthermore, the molar ratio of the glycosyl donor to the glycosyl acceptor in the synthetic route is (1-2):1; the glycosidation reaction conditions are as follows: dissolving the glycosyl donor and the glycosyl acceptor in a dry solvent, adding molecular sieves and a promoter, and stirring the reaction at a set temperature for 3-7 hours to obtain the target oligosaccharide fragment; the dry solvent is at least one of anhydrous dichloromethane, anhydrous ether, anhydrous toluene, anhydrous methanol, anhydrous tetrahydrofuran, anhydrous acetonitrile, anhydrous N,N-dimethylformamide, and anhydrous dioxane; and the stirring at the set temperature is stirring the reaction at room temperature of 25°C, or stirring the reaction in an ice-water mixture at 0°C, or stirring the reaction in a mixture of ice and sodium chloride at -5 to -20°C, or stirring the reaction in a mixture of acetonitrile and dry ice at -40°C, or stirring the reaction in a mixture of acetone and dry ice at -60°C, or stirring the reaction in a mixture of acetone and dry ice at -78°C.

[0032] A pharmaceutical composition comprises the lycium ruthenicum oligosaccharide derivative, a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0033] A use of a pharmaceutical composition in preparing a drug for preventing or treating pancreatic cancer.

[0034] The advantages of the present invention are as follows: the present invention utilizes the remote participation effect of acyl groups, steric hindrance effect, solvent effect and end group effect to realize the efficient construction of 1,2-cis-α-galacturonic acid glycosidic bonds and other glycosidic bonds in the oligosaccharide structure, thereby realizing the first chemical synthesis of black wolfberry oligosaccharides and derivatives thereof; the black wolfberry oligosaccharide fragments provided by the present invention have an inhibitory effect on pancreatic cancer cells, provide a theoretical basis for clarifying the active fragments (active structural domains) of black wolfberry polysaccharides, for conducting more in-depth functional mechanism research and for developing new anti-pancreatic cancer drugs based on the active fragments thereof, and have good application prospects in the preparation of new anti-cancer drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 : Effects of synthetic Lycium barbarum oligosaccharide fragments and gemcitabine on the growth of pancreatic cancer cells PANC-1.

[0036] Figure 2 : Effects of synthetic Lycium barbarum oligosaccharide fragments and gemcitabine on the growth of pancreatic cancer cells AsPC-1.

[0037] Figure 3 : Effects of synthetic Lycium barbarum oligosaccharide fragments and gemcitabine on the growth of pancreatic cancer cells BxPC-3.

[0038] Figure 4 :IC of disaccharide 44 on pancreatic cancer cells PANC-1 50 Test relationship diagram. DETAILED DESCRIPTION

[0039] Example

[0040] A lycium barbarum oligosaccharide derivative, wherein the derivative is a compound having a structure represented by formula (1) to (8) or a pharmaceutically acceptable salt thereof:

[0041]

[0042] Among them, PG1, PG5, PG 10 PG 17 is any one of hydroxy, acetoxy, benzoyloxy, pivaloyloxy, chloroacetoxy, levulinyloxy, allyloxycarbonyloxy, methoxy, benzyloxy, 2-naphthylmethoxy, p-methoxybenzyloxy, allyloxy, p-methoxyphenoxy, trityloxy, monomethoxytrityloxy, dimethoxytrityloxy, tert-butyldimethylsilyloxy, tert-butyldiphenylsilyloxy, triethylsilyloxy, halogen, trichloroacetimidyloxy, N-phenyltrifluoroacetimidyloxy, dibenzylphosphoxy, o-alkynylbenzoyloxy, o-methoxycarbonylalkynylphenylthio, methylthio, ethylthio, phenylthio, and p-tolylthio; PG2, PG6, PG 11 PG 14 It can be any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, levulinyl, and allyloxycarbonyl; PG3, PG4, PG7, PG8, PG9, PG 12 PG 15 PG 16 PG is any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, levulinyl, allyloxycarbonyl, methyl, benzyl, 2-naphthylmethyl, p-methoxybenzyl, allyl, p-methoxyphenyl, trityl, monomethoxytrityl, dimethoxytrityl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, triethylsilyl, benzylideneacetal, and isopropylideneketal; 13 Any one of hydrogen, methyl, ethyl, tert-butyl, benzyl, allyl, trichloroethyl, and trimethylsilylethyl; PG 18 It is any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, levulinyl and allyloxycarbonyl.

[0043] Furthermore, the pharmaceutically acceptable salt is any one of sodium salt, potassium salt, calcium salt, magnesium salt, zinc salt and meglumine salt.

[0044] A method for preparing a lycium barbarum oligosaccharide derivative comprises the following steps:

[0045] S1. Glycosylation of glycosyl donor 9 and glycosyl acceptor 10 yields disaccharide 1. Acyl protecting groups of disaccharide 1 are removed under alkaline conditions, followed by reduction to remove aromatic protecting groups, completing deprotection and yielding fully deprotected disaccharide 1a. The synthetic route is as follows:

[0046]

[0047] wherein LG is a leaving group of the glycosyl donor and is any one of halogen, trichloroacetimidyloxy, N-phenyltrifluoroacetimidyloxy, dibenzylphosphoxy, o-alkynylbenzoyloxy, o-methoxycarbonylalkynylphenylthio, methylthio, ethylthio, phenylthio, and p-tolylthio;

[0048] S2. Disaccharide 1 is selectively deprotected by PG4 to yield disaccharide 11, which is then glycosylated with glycosyl donor 9 under the catalytic action of a promoter to yield trisaccharide 2. Trisaccharide 2 is then deprotected by removing the acyl protecting group under alkaline conditions and the aromatic protecting group by reduction, completing deprotection and yielding fully deprotected trisaccharide 2a. The synthetic route is shown below:

[0049]

[0050] S3. Glycosidation of glycosyl donor 9 and glycosyl acceptor 12 yields disaccharide 3. Acyl protecting groups of disaccharide 3 are removed under alkaline conditions, followed by reduction to remove aromatic protecting groups, completing deprotection and yielding fully deprotected trisaccharide 3a. The synthetic route is as follows:

[0051]

[0052] S4. Glycosylation of glycosyl donor 13 and glycosyl acceptor 14 yields disaccharide 4. Acyl protecting groups of disaccharide 4 are removed under alkaline conditions, followed by reduction to remove aromatic protecting groups, completing deprotection and yielding fully deprotected trisaccharide 4a. The synthetic route is as follows:

[0053]

[0054] S5. Glycosylation of glycosyl donor 13 and glycosyl acceptor 15 yields disaccharide 5. Acyl protecting groups of disaccharide 5 are removed under alkaline conditions, followed by reduction to remove aromatic protecting groups, completing deprotection and yielding fully deprotected trisaccharide 5a. The synthetic route is as follows:

[0055]

[0056] S6. Glycosylation of glycosyl donor 16 and glycosyl acceptor 14 yields disaccharide 6. The acyl protecting group of disaccharide 6 is removed under alkaline conditions, and the aromatic protecting group is removed by reduction to complete deprotection, yielding fully deprotected trisaccharide 6a. The synthetic route is as follows:

[0057]

[0058] S7. Glycosylation of glycosyl donor 17 and glycosyl acceptor 18 is performed, utilizing the remote participation effect of the C4 acyl group to complete the construction of a 1,2-cis-glycosidic bond, yielding disaccharide 7. Disaccharide 7 is then deprotected by removing the acyl protecting group under alkaline conditions, followed by reduction to remove the aromatic protecting group, yielding fully deprotected disaccharide 7a. The synthetic route is as follows:

[0059]

[0060] S8. Glycosylation of glycosyl donor 19 and glycosyl acceptor 14 yields trisaccharide 8. Trisaccharide 7 is deprotected by removing the acyl protecting group under alkaline conditions, followed by reduction to remove the aromatic protecting group, thereby completing deprotection and yielding fully deprotected trisaccharide 8a. The synthetic route is as follows:

[0061]

[0062] Furthermore, the accelerator is any one of boron trifluoride etherate, trifluoromethanesulfonic acid, trimethylsilyl trifluoromethanesulfonate, silver carbonate, trifluoromethanesulfonic acid, N-iodosuccinimide and trifluoromethanesulfonic acid, N-iodosuccinimide and N-iodosuccinimide and trifluoromethanesulfonic acid, triphenylphosphine gold trifluoromethanesulfonate, and triphenylphosphine gold bis(fluorosulfonyl)imide.

[0063] Furthermore, the reduction reaction conditions for removing the aromatic protecting group are: a mixed solvent consisting of dichloromethane or tetrahydrofuran or ethyl acetate and tert-butanol or methanol, water, and acetic acid, a 10% palladium carbon catalyst or palladium hydroxide, and hydrogen, and stirring at room temperature for 48 hours; or sodium, liquid ammonia, tetrahydrofuran, and tert-butanol, and stirring at -78°C for 30 minutes.

[0064] Furthermore, the molar ratio of the glycosyl donor to the glycosyl acceptor in the synthetic route is (1-2):1; the glycosidation reaction conditions are as follows: dissolving the glycosyl donor and the glycosyl acceptor in a dry solvent, adding molecular sieves and a promoter, and stirring the reaction at a set temperature for 3-7 hours to obtain the target oligosaccharide fragment; the dry solvent is at least one of anhydrous dichloromethane, anhydrous ether, anhydrous toluene, anhydrous methanol, anhydrous tetrahydrofuran, anhydrous acetonitrile, anhydrous N,N-dimethylformamide, and anhydrous dioxane; and the stirring at the set temperature is stirring the reaction at room temperature of 25°C, or stirring the reaction in an ice-water mixture at 0°C, or stirring the reaction in a mixture of ice and sodium chloride at -5 to -20°C, or stirring the reaction in a mixture of acetonitrile and dry ice at -40°C, or stirring the reaction in a mixture of acetone and dry ice at -60°C, or stirring the reaction in a mixture of acetone and dry ice at -78°C.

[0065] A pharmaceutical composition comprises the lycium ruthenicum oligosaccharide derivative, a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0066] A use of a pharmaceutical composition in preparing a drug for preventing or treating pancreatic cancer.

[0067] Experimental example

[0068] General experimental methods

[0069] The anhydrous solvents used in the experiment were commercial reagents (Annaiji Chemical). 1 H NMR, 13 C NMR, 1 H- 1 H COSY, 1 H- 13 C HSQC measurements were performed on a Bruker AVANCEШ500 NMR spectrometer using TMS as the internal standard at 25°C. Peak types were represented by singlets (s), doublets (d), triplets (t), quartets (dd), and multiplets (m). Chemical shifts (δ) were reported in ppm, and coupling constants (J) were reported in Hz. Mass spectra were determined on a Thermo Scientific Ultimate 3000 Orbitrap Exploris 240 ultra-high performance liquid chromatography-mass spectrometer in positive and negative ion full scan mode. Column chromatography and thin-layer chromatography (TLC) were performed on 200-300 mesh silica gel and GF254 silica gel plates, manufactured by Shandong Qingdao Ocean Chemical Plant. Color development was performed using a 5% (v / v) sulfuric acid-ethanol solution heated with a heat gun and under ultraviolet light.

[0070] Glycosylation general method A: Combine the glycosyl donor (1.5 equivalents) and the glycosyl acceptor (1.0 equivalents), dissolve in toluene, and spin dry to remove water twice (temperature 30°C). Under nitrogen protection, the above raw materials are dissolved in anhydrous dichloromethane (reaction concentration 0.1-0.5 mol / L), add molecular sieves, stir at -45°C for 10 minutes, add TMSOTf (0.3 equivalents), stir and react at -45°C for 2 hours. After the reaction is completed by TLC monitoring, the reaction solution is transferred to dichloromethane and extracted with dichloromethane and saturated sodium bicarbonate. The organic phase is dried over anhydrous sodium sulfate and spin dried. It is separated and purified by medium-pressure semi-preparative chromatography to obtain the target oligosaccharide.

[0071] General method B for glycosylation: The glycosyl donor (1.5 equivalents) and the glycosyl acceptor (1.0 equivalents) were combined and azeotroped with toluene twice. Under nitrogen protection, the mixture was dissolved in 5 mL of DCM, and molecular sieves and NIS (2.0 equivalents) were added. TMSOTf (0.2 equivalents) was added at 0°C, and the reaction was stirred at 0°C for 5 h. After the reaction was completed, the reaction solution was transferred to DCM, filtered, extracted with DCM / saturated sodium thiosulfate, and extracted with saturated NaHCO3 solution. The organic phase was dried over anhydrous sodium sulfate, concentrated and dried, and purified by column chromatography to obtain the target oligosaccharide.

[0072] General procedure for the hydrolysis of 4-methoxyphenyl: Dissolve the compound (1.0 equiv) in DCM / MeCN / H₂O (reaction concentration 0.1-0.5 mol / L, 2 / 1 / 2, v / v / v), add CAN (4.0 equiv), and react at room temperature for 5 h. After completion of the reaction as monitored by TLC, transfer the reaction mixture to DCM and extract with DCM / 1M hydrochloric acid and saturated sodium bicarbonate. The organic phase is dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by medium-pressure semi-preparative chromatography to yield the title compound.

[0073] General method for removal of silyl ether protecting groups: Compound 25 (1.0 equivalent) was dissolved in tetrahydrofuran (reaction concentration 0.1-0.5 mol / L), and acetic acid (1.5 equivalent) and TBAF (1.5 equivalent) were added at 0°C. The mixture was reacted in an ice bath for 12 h. After TLC detection showed that the reaction of the raw materials was complete, dichloromethane was added to dilute the reaction solution, and the mixture was extracted with dichloromethane, saturated sodium bicarbonate solution, and saturated brine. The organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated and purified by silica gel column chromatography to obtain the title compound.

[0074] General method for glucosinolate hydrolysis: The compound (1.0 equivalent) was dissolved in acetone / water (reaction concentration 0.1 mol / L, 4 / 1, v / v), trichloroisocyanuric acid (1.7 equivalent) was added, and the reaction was carried out at 0°C for 5 minutes. After TLC detection of the complete reaction of the raw material, dichloromethane was added to dilute the reaction solution, and the solution was extracted with dichloromethane, saturated sodium bicarbonate solution, and saturated brine. The organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by semi-preparative chromatography to obtain a compound with a hydroxyl group at position 1.

[0075] General method for acyl removal: The compound (1.0 equivalent) was dissolved in methanol / dichloromethane solution (reaction concentration 0.1 mol / L, 1 / 1, v / v) and sodium methoxide (0.2 equivalent / each acyl group). After reacting at room temperature for 2 hours, hydrogen ion exchange resin was added to pH = 7. The mixture was filtered and concentrated under reduced pressure. The target compound was separated and purified by silica gel chromatography to obtain the target compound.

[0076] General catalytic hydrogenation procedure: Dissolve the compound (1.0 equivalent) in a mixture of chromatographic-grade methanol / dichloromethane / water (2 mL / 2 mL / 0.2 mL, 10 / 10 / 1, v / v / v). Add excess palladium on carbon, replace the atmosphere with hydrogen for 20 minutes, and stir under hydrogen at room temperature for 2 days. Transfer the reaction mixture to distilled water, filter, concentrate, and purify using a Sep-Pak C18 cartridge. Collect the aqueous phase to obtain the fully deprotected oligosaccharide.

[0077] Experimental Example 1

[0078] The synthesis of arabinose building blocks 26 and 29 was carried out as follows:

[0079]

[0080] Specific test operations and steps:

[0081] Compound 21: Compound 20 (1.058 g, 4.12 mmol) was dissolved in dimethylformamide (20 mL), and the reaction system was placed in a low-temperature reaction zone at -20°C. 2,6-Lutidine (1.750 mL, 10.30 mmol) and di-tert-butylbis(trifluoromethanesulfonyloxy)silane (2.20 mL, 4.53 mmol) were added, and the reaction was carried out at -20°C for 2.5 hours. After TLC detection showed that the reaction of the raw materials was complete, methanol was added to quench the reaction. The reaction solution was extracted with dichloromethane / saturated sodium bicarbonate solution, the filtrate was dried over anhydrous sodium sulfate, and the organic layer was concentrated under reduced pressure. The mixture was separated and purified by silica gel column chromatography (PE / EA=10 / 1) to obtain compound 21 (1.153 g, 2.90 mmol, 71%) as a white solid. 1 H NMR (500MHz, CDCl3) δ = 7.44-7.39 (m, 2H, Ar), 7.12 (d, J = 7.8Hz, 2H, Ar), 5.24 ( d,J=5.9Hz,1H,H-1),4.35-4.30(m,1H,H-5),4.13(dd,J=7.5,5.9Hz,1H,H-2) ,4.00(dd,J=9.2,7.4Hz,1H,H-3),3.94(dd,J=10.3,8.8Hz,1H,H-5),3.90-3. 85(m,1H,H-4),2.32(s,3H,STol-CH3),1.06(s,9H,t-Bu),0.97(s,9H,t-Bu).

[0082] Compound 22: Compound 21 (1.13 g, 2.85 mmol) was dissolved in pyridine (20 mL), and BzCl (1.0 ml, 8.6 mmol) was added at 0°C. The reaction was allowed to react overnight. After TLC detection showed that the reaction of the raw materials was complete, dichloromethane was added to dilute the reaction solution. The solution was extracted with dichloromethane / saturated sodium bicarbonate solution and saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The product was separated and purified by silica gel column chromatography (PE / EA=30 / 1) to give the target compound 22 (1.38 g, 15.24 mmol, 97%). 1 H NMR (500MHz, CDCl3) δ = 8.09 (dd, J = 8.2, 1.4Hz, 2H, Ar), 7.65-7.56 (m, 1H, Ar), 7.52-7.40 ( m,4H,Ar),7.12(d,J=7.9Hz,2H,Ar),5.51(dd,J=7.0,4.7Hz,1H,H-2),5.37(d,J=4.7Hz,1H ,H-1),4.39(dd,J=8.9,4.8Hz,1H,H-5),4.32-4.28(m,1H,H-4),4.07(td,J=9.9,4.8Hz,1 H,H-5),4.01(d,J=9.0Hz,1H,H-3),2.31(s,3H,STol-CH3),1.01(d,J=28.8Hz,18H,t-Bu).

[0083] Compound 23: Compound 22 (7.873 g, 15.7 mmol) was treated with a general method for removing the silyl ether protecting group to obtain compound 23 (5.65 g, 15.7 mmol, quant.) as a light yellow syrup. 1 H NMR(500MHz, CDCl3)δ=8.06-7.99(m,2H,Ar),7.63-7.56(m,1H,Ar),7.46(t, J=7.5Hz,4H,Ar),7.15(d,J=7.9Hz,2H,Ar),5.68(d,J=3.2Hz,1H,H-2),5.14( t,J=3.5Hz,1H,H-1),4.31(ddd,J=27.8,7.5,3.8Hz,2H,H-5),3.96(dd,J=12. 4,2.9Hz,1H,H-4),3.81(dd,J=12.4,3.8Hz,1H,H-3),2.34(s,3H,STol-CH3).

[0084] Compound 24: Compound 23 (490 mg, 1.36 mmol) was dissolved in dichloromethane (15 mL), and imidazole (232 mg, 3.4 mmol) and tert-butyldimethylsilyl chloride (308 mg, 2.04 mmol) were added at 0°C. The mixture was allowed to react at room temperature for 2 hours. After TLC analysis showed that the reaction of the starting materials was complete, dichloromethane was added to dilute the reaction solution. The reaction solution was extracted with dichloromethane, saturated sodium bicarbonate solution, and saturated brine. The organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated and purified by silica gel column chromatography (PE / EA = 10 / 1) to obtain the target compound 24 (450 mg, 1.02 mmol, 75%). 1 H NMR (400MHz, CDCl3) δ = 7.59 (ddt, J = 7.9, 7.0, 1.4Hz, 1H, Ar), 7.49-7.40 (m, 4H, Ar), 7.16-7.09(m,2H,Ar),5.65(d,J=3.7Hz,1H,H-1),5.14(t,J=3.6Hz,1H,H-2),4.28( dt,J=6.6,3.3Hz,2H,H-3,H-4),3.89(qd,J=11.4,3.5Hz,2H,H-5),3.42(s,1H,H-2) ,2.33(s,3H,STol-CH3),0.87(s,9H,TBDMS-CH3),0.06(d,J=2.0Hz,6H,TBDMS-CH3). 13 C NMR(101MHz, CDCl3)δ=167.2,137.8,133.7,132.6,129.9,129.8,129.7,128.9 ,128.5,89.2,86.9,83.1,77.3,77.2,77.0,76.8,76.7,62.5,25.8,21.1,18.5.

[0085] Compound 25: Compound 24 (4.150 g, 8.74 mmol) was dissolved in dichloromethane (90 mL), and silver oxide (6.076 g, 26.22 mmol) was added. Benzyl bromide (3.6 mL, 30.59 mmol) was added at 0°C, and the mixture was reacted at room temperature for 30 h. After TLC detection showed that the reaction of the raw materials was complete, dichloromethane was added to dilute the reaction solution. The reaction solution was filtered through celite and extracted with dichloromethane, saturated sodium bicarbonate solution, and saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The mixture was separated and purified by silica gel column chromatography (PE / EA=30 / 1) to give compound 25 (3.407 g, 6.03 mmol, 69%) as a transparent syrup. 1H NMR(500MHz, CDCl3)δ=8.03-7.98(m,2H,Ar),7.61-7.54(m,1H,Ar),7.47-7.40(m,4H,Ar),7.38-7.25(m,6 H,Ar),7.10(d,J=7.9Hz,2H,Ar),5.60(d,J=1.6Hz,1H,H-1),5.57(d,J=1.8Hz,1H,H-2),4.80(d,J=12.0Hz ,1H,Ar-CH2),4.64(d,J=12.0Hz,1H,Ar-CH2),4.44(q,J=4.7Hz,1H,H-4),4.14(dd,J=5.5,1.9Hz,1H,H-3) ,3.80(d,J=4.5Hz,2H,H-5),2.31(s,3H,STol-CH3),0.84(s,9H,TBS-CH3),0.00(d,J=4.4Hz,6H,TBS-CH3). 13 C NMR (101MHz, CDCl3) δ=165.4,137.6,137.6,133.4,132.5,130.7,129.8,129.7,129.4,128.5,128.4,127.9,12 7.8,91.5,83.5,82.9,82.5,77.4,77.2,77.0,76.7,72.2,62.3,31.6,29.7,29.7,25.9,21.1,18.3,-5.3,-5.4.

[0086] Compound 26: Compound 25 (510 mg, 0.90 mmol) was subjected to a general method for removing silyl ether protecting groups to obtain compound 26 (396 mg, 0.88 mmol, 98%) as a clear syrup. 1HNMR (500MHz, CDCl3) δ = 8.06-7.97 (m, 2H, Ar), 7.58 (t, J = 7.4Hz, 1H, Ar), 7.44 (dt, J = 7.8, 3.7Hz, 4H, Ar), 7.32 (ddd,J=26.8,18.5,7.1Hz,5H,Ar),7.12(d,J=7.8Hz,2H,Ar),5.61(s,1H,H1),5.56(d,J=1.7Hz,1H,H-2),4.8 4(d,J=11.9Hz,1H,Ar-CH2),4.63(d,J=11.9Hz,1H,Ar-CH2),4.49(dt,J=6.2,3.4Hz,1H,H-4),4.16(dd,J=5.6 ,1.8Hz,1H,H-3),3.89(dd,J=12.3,3.0Hz,1H,H-5),3.70(dd,J=12.2,3.9Hz,1H,H-5),2.32(s,3H,STol-CH3).

[0087] Compound 27: Compound 25 (840 mg, 1.49 mmol) was subjected to the general method for glucosinolate hydrolysis to obtain compound 27 (594 mg, 1.3 mmol, 87%) as a clear syrup.

[0088] Compound 28: Compound 27 (157 mg, 0.34 mmol) and o-cyclopropylethynylbenzoic acid (95 mg, 0.51 mmol) were dissolved in 5 mL of ultra-dry DCM. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (117 mg, 0.612 mmol) and 4-dimethylaminopyridine (62 mg, 0.51 mmol) were added. The mixture was allowed to react at room temperature for 1.5 hours. After TLC analysis, the reaction was complete, and the mixture was concentrated under reduced pressure. The mixture was separated and purified by silica gel column chromatography (PE / EA = 50 / 1) to obtain compound 28 (170 mg, 0.27 mmol, 80%, α / β = 10 / 7) as a clear syrup. 1H NMR (500MHz, CDCl3) δ = 8.06 (d, J = 7.4Hz, 2H, Ar), 7.95 (dd, J = 8.2, 3.4Hz, 1H, Ar), 7.61 (t, J = 7.4Hz, 1H, Ar), 7.51-7.27 (m, 9H, Ar) ),7.22(t,J=7.6Hz,1H,Ar),6.60(d,J=3.6Hz,1H,H-1),5.67(d,J=3.3Hz,1H,H-2),4.87(dd,J=11.9,3.5Hz,1H,Ar-CH2),4.69( dd,J=11.8,3.6Hz,1H,Ar-CH2),4.44(p,J=4.5Hz,1H,H-3),4.24(d,J=4.3Hz,1H,H-4),3.84(dd,J=11.1,4.3Hz,1H,H-5),3.78( dd,J=11.0,5.5Hz,1H,H-5),1.58-1.49(m,1H,ABz-CH),0.90-0.81(m,13H,ABz-CH2,TBS-CH3),0.02(t,J=5.3Hz,6H,TBS-CH3).

[0089] Compound 29: Compound 28α (54 mg, 0.086 mmol) was dissolved in a mixed solution of methanol / dichloromethane (1.2 mL / 0.3 mL, 1 / 4, v / v), and acetyl chloride (60 μL, 0.86 mmol) was added. The reaction was allowed to react at room temperature for 45 minutes. After TLC detection showed that the reaction of the raw materials was complete, dichloromethane was added to dilute the reaction solution. The solution was extracted with dichloromethane, saturated sodium bicarbonate solution, and saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The product was separated and purified by silica gel column chromatography (PE / EA=8 / 1) to give compound 29 (37.6 mg, 73.1 μmol, 85%) as a transparent syrup. 1HNMR(500MHz, CDCl3)δ=8.07-8.03(m,2H,Ar),7.96(dd,J=8.0,1.4Hz,1H,Ar),7.63-7.59(m,1H,Ar),7.50-7.41(m,4H,Ar),7 .37-7.34(m,2H,Ar),7.33-7.27(m,3H,Ar),7.24(dd,J=7.6,1.3Hz,1H,Ar),6.59(s,1H,H-1),5.69(d,J=1.6Hz,1H,H-2),4.9 0(d,J=11.7Hz,1H,Ar-CH2),4.67(d,J=11.8Hz,1H,Ar-CH2),4.48(dt,J=5.0,3.5Hz,1H,H-4),4.24(dt,J=5.3,1.2Hz,1H,H-3 ),3.92(dd,J=12.4,3.0Hz,1H,H-5),3.76-3.69(m,1H,H-5),1.53(tt,J=7.8,5.3Hz,1H,ABz-CH),0.88-0.84(m,4H,ABz-CH2).

[0090] Experimental Example 2

[0091] The synthesis of galacturonic acid sugar building block 32 is as follows:

[0092]

[0093] Compound 31: Compound 30 (3.296 g, 6.67 mmol) was dissolved in dichloromethane (34 mL), and dicyclohexylcarbodiimide (2.75 g, 13.3 mmol), levulinic acid (1.36 mL, 13.3 mmol), and 4-dimethylaminopyridine (1.625 g, 13.3 mmol) were added sequentially. The mixture was allowed to react at room temperature for 24 hours. After completion of the reaction as monitored by TLC, the reaction solution was extracted with dichloromethane, saturated sodium bicarbonate solution, and saturated aqueous sodium chloride solution. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The mixture was separated and purified by preparative chromatography (PE / EA = 7 / 3) to give compound 31 (3.925 g, 6.62 mmol, 99%).

[0094] Compound 32: Compound 31 (711.5 mg, 1.20 mmol) was hydrolyzed using the general method for 4-methoxyphenyl to obtain a hemiacetal intermediate (446.1 mg, 0.92 mmol, 76%). The hemiacetal intermediate (904 mg, 1.525 mmol) was dried under vacuum and dissolved in anhydrous dichloromethane (10 mL). 2,2,2-Trifluoro-N-phenylacetimidoacetyl chloride (1.2 mL, 7.6 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (0.68 mL, 4.575 mmol) were added at 0°C and allowed to react at room temperature for 2 hours. After completion of the reaction by TLC, the reaction system was concentrated under reduced pressure at 30°C and purified by medium-pressure semi-preparative chromatography (PE / EA = 5 / 1) to obtain compound 32 (1.044 g, 1.5 mmol, quant.).

[0095] Experimental Example 3

[0096] The synthesis of rhamnose building blocks 35 and 38 is as follows:

[0097]

[0098] Compound 34: Compound 33 (6.63 g, 21.36 mmol) was dissolved in dimethylformamide (107 mL). Sodium hydride (1.71 g, 42.8 mmol) was added at 0°C, followed by 2-bromomethylnaphthalene (9.45 g, 42.8 mmol). The mixture was stirred at room temperature. The reaction was completed by TLC. Distilled water was added to terminate the reaction in an ice-water bath. The mixture was extracted with distilled water and ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the intermediate compound. The intermediate compound was dissolved in 80% aqueous acetic acid (25 mL), heated under reflux at 60°C, and reacted for 5 hours. The reaction solution was concentrated under reduced pressure to obtain the intermediate compound. The intermediate compound was dissolved in toluene (200 mL), dibutyltin oxide (11.65 g, 46.8 mmol) was added, and the mixture was heated under reflux at 120°C for 4 hours. The mixture was returned to room temperature, half of the solvent was removed by vortexing, and acetonitrile (100 mL) was added, followed by cesium fluoride (7.109 g, 46.8 mmol) and benzyl bromide (8.0 g, 46.8 mmol). The mixture was heated under reflux at 80°C for 12 h. The reaction was confirmed by TLC, and the mixture was transferred to dichloromethane and filtered through Celite to remove insoluble impurities. The mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA = 8 / 1) to obtain compound 34 (6.35 g, 13.05 mmol, 61%). 1H NMR(500MHz, CDCl3)δ=7.87-7.72(m,4H,Ar),7.52-7.41(m,4H,Ar),7.40-7.28(m,8H,A r),7.10(d,J=7.8Hz,2H,Ar),5.46(s,1H,H-1),5.04(d,J=11.2Hz,1H,Ar-CH2),4.81(d, J=11.1Hz,1H,Ar-CH2),4.72(s,2H,Ar-CH2),4.28-4.20(m,2H,H-2,H-5),3.90(dd,J=9. 1,3.2Hz,1H,H-3),3.58(t,J=9.3Hz,1H,H-4),2.31(s,3H,STol-CH3),1.32(s,3H,H-6). 13 C NMR(101MHz, CDCl3)δ=137.6,132.1,129.9,128.7,128.2,128.1,128.0,127.9,127.7, 126.6,126.1,126.0,125.9,87.4,80.2,80.1,77.3,75.5,72.2,70.1,68.7,21.1,17.9.

[0099] Compound 35: Under nitrogen, compound 34 (4.3 g, 8.8 mmol) was dissolved in pyridine (95 ml), and acetic anhydride (1.65 mL, 17.6 mmol) and 4-dimethylaminopyridine (215 mg, 1.76 mmol) were added at 0°C. The mixture was stirred at 0°C for 0.5 hours and then allowed to react at room temperature for 2 hours. The reaction was confirmed to be complete by TLC. The reaction system was transferred to ethyl acetate and extracted with 1 M hydrochloric acid and saturated sodium bicarbonate, respectively. The organic phase was dried over anhydrous sodium sulfate, concentrated and dried, and purified by diatom column chromatography (PE / EA = 15 / 1) to obtain compound 35 (4.64 g, 8.7 mmol, 99%). 1HNMR (500MHz, CDCl3) δ=7.77-7.68(m,3H),7.63(s,1H,Ar),7.39(tt,J=6.6,3.7Hz,2H,Ar),7.33(dd,J=7.9,5.8Hz,3H ,Ar),7.29-7.25(m,2H,Ar),7.25-7.16(m,4H,Ar),7.04(d,J=7.8Hz,2H,Ar),5.73(d,J=3.3Hz,1H,H-2),4.97(d,J=11. 0Hz,1H,Ar-CH2),4.76-4.67(m,3H,H1,Ar-CH2),4.42(d,J=11.1Hz,1H,Ar-CH2),3.57(dd,J=9.2,3.4Hz,1H,H-3),3.43 (t,J=9.2Hz,1H,H-4),3.38-3.29(m,1H,H-5),2.25(s,3H,STol-CH3),2.16(s,3H,Ac-CH3),1.34(d,J=6.1Hz,3H,H-6).

[0100] Compound 37: Under nitrogen, compound 36 (595.9 mg, 2.44 mmol) was dissolved in DMF (12 mL). Sodium hydride (195.2 mg, 4.88 mmol) was added under an ice-water bath, followed by 2-bromomethylnaphthalene (1.08 g, 4.88 mmol). The ice-water bath was removed and the reaction was stirred at room temperature. The reaction was completed by TLC. Distilled water was added to terminate the reaction under an ice-water bath. The mixture was extracted with distilled water and ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the intermediate compound. The mixture was then dissolved in 80% aqueous acetic acid (25 mL), heated under reflux at 60°C, and reacted for 5 hours. The reaction system was extracted with dichloromethane and water. The organic phase was filtered over anhydrous sodium sulfate, concentrated, and purified on a silica gel column (PE / EA = 4 / 1) to obtain compound 37 (782.4 mg, 2.27 mmol, 93%).

[0101] Compound 38: Compound 37 (782.4 mg, 2.27 mmol) was dissolved in toluene (12 ml), and dibutyltin oxide (1.13 g, 4.54 mmol) was added. The mixture was heated under reflux at 120°C for 5 h. After returning to room temperature, half of the solvent was removed by vortexing, and acetonitrile (6 mL) was added. Benzyl bromide (539 μL, 4.54 mmol) and tetrabutylammonium bromide (1.46 g, 4.54 mmol) were then added in that order. The mixture was heated under reflux at 80°C for 12 h. The reaction was confirmed by TLC, and the mixture was transferred to dichloromethane. Insoluble impurities were removed by filtration through celite, and the mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA = 7 / 1) to afford compound 38 (768 mg, 1.77 mmol, 78%).

[0102] Experimental Example 4

[0103] The synthesis routes of Lycium barbarum oligosaccharides 41, 44, 47, 51 and 53 are as follows:

[0104]

[0105] Compound 40: Glycosyl donor 39 (241 mg, 0.4 mmol) and acceptor 26 (130 mg, 0.29 mmol) were glycosylated using General Glycosylation Method A to afford compound 40 (144 mg, 0.17 mmol, 59%). 1 HNMR (500MHz, CDCl3) δ = 8.03 (dd, J = 8.3, 1.3Hz, 2H, Ar), 7.99-7.92 (m, 2H, Ar), 7.60-7 .52(m,2H,Ar),7.47-7.33(m,8H,Ar),7.30-7.27(m,2H,Ar),7.25-7.18(m,10H,Ar),7. 08(d,J=7.9Hz,2H,Ar),5.66-5.60(m,1H,A-H1),5.57(t,J=1.8Hz,1H,A-H2),5.38(d,J =1.5Hz,1H,D-H2),5.27(s,1H,D-H1),4.79(d,J=11.9Hz,1H,Ar-CH2),4.65-4.61(m,2H ,Ar-CH2),4.59(q,J=4.3Hz,1H,D-H3),4.53(d,J=12.1Hz,1H,Ar-CH2),4.46(dd,J=12 .2,4.6Hz,2H,Ar-CH2),4.27(ddd,J=5.4,2.0,0.9Hz,1H,A-H3),4.20(td,J=5.2,3.7Hz ,1H,A-H4),4.01-3.90(m,2H,D-H4,D-H5),3.78(dd,J=11.4,4.1Hz,1H,D-H5),3.59(dd ,J=10.7,3.7Hz,1H,A-H5),3.52(dd,J=10.8,5.0Hz,1H,A-H5),2.29(s,3H,STol-CH3). 13 CNMR(126MHz, CDCl3)δ=137.9,137.6,133.3,132.4,130.0,129.6,129.4,128.6,128 .1,128.0,127.4,106.2,91.4,83.2,82.3,81.6,77.2,73.4,72.4,72.1,69.3,65.4.

[0106] Compound 41: Compound 40 (128 mg, 0.15 mmol) was subjected to the general method for glucosinolate hydrolysis, the general method for acyl removal and the general method for catalytic hydrogenation to obtain compound 41 (7 mg, 0.042 mmol, 29%). 1 HNMR(600MHz,D2O)δ=5.07(d,J=3.6Hz,0.6H),4.90(s,2H),4.81(s,0.4H),4.35(d,J=7.8Hz,1H),3.98-3.96(m,2H),3.93-3.91 (m,2H),3.78(s,2H),3.72(s,2H),3.66(s,3H),3.57(d,J=4.4Hz,2H),3.54(s,2H),3.41(d,J=8.3Hz,2H),3.35(d,J=8.7Hz,1H). 13 C NMR(151MHz,D2O)δ=107.4,96.8,92.6,83.9,80.9,76.6,72.4,72.2,71.9,70.9,7 0.5,70.2,69.3,68.8,68.6,68.1,66.4,63.0,62.5,61.2.HR-ESI-MS(m / z):calcd for C 10 H 18 O9Na + (M+Na) + :305.0843,found:305.0822.

[0107] Compound 43: Glycosyl donor 39 (140 mg, 0.23 mmol) and acceptor 42 (59 mg, 0.116 mmol) were glycosylated according to general method A to give compound 87 (85 mg, 0.09 mmol, 78%). 1HNMR(500MHz, CDCl3)δ=8.01-7.95(m,2H,Ar),7.60-7.53(m,2H,Ar),7.49(tt,J=7.5,1.4Hz,1H,Ar),7.23-7.15(m,9H,Ar),5.76(t,J=9.8H z,1H,A-H2),5.37(s,1H,D-H2),5.29(d,J=1.3Hz,1H,D-H1),5.08(d,J=11.7Hz,1H,Ar-CH2),4.67(d,J=11.9Hz,1H,Ar-CH2),4.59-4.40(m, 8H, Ar-CH2, A-H1, D-H3), 4.12 (dd, J=9.7, 2.8Hz, 1H, A-H3), 4.00 (ddd, J=7.8, 3.3, 1.0Hz, 1H, A-H4), 3.94 (dt, J=5.0, 1.0Hz, 1H, D-H5), 3.76 (ddd,J=6.8,5.5,1.1Hz,1H,D-H5),3.67-3.54(m,4H,D-H4,A-H5,A-H6),2.80-2.64(m,2H,SEt-CH2),1.22(dt,J=18.6,7.5Hz,3H,SEt-CH3). 13 CNMR(126MHz, CDCl3)δ=164.9,137.9,133.1,132.7,129.9,129.7,128.7,127.8,127.7,127.3, 107.7,83.8,83.4,81.3,79.5,77.9,77.3,76.2,74.8,73.5,72.3,71.0,69.5,68.8,23.8,14.9.

[0108] Compound 44: Compound 43 (34 mg, 0.037 mmol) was sequentially subjected to the general method for glucosinolate hydrolysis, the general method for acyl removal and the general method for catalytic hydrogenation to obtain compound 44 (12 mg, 0.0178 mmol, 48%). 1 HNMR(600MHz,D2O)δ5.12(d,0.4H),5.08(dd,J=4.6,1.6Hz,1H),4.48(d,J=7.9Hz,1H),4.05(dt,J=4.0,2.0Hz,1H),4.01 -3.93(m,2.8H),3.81-3.77(m,1.2H),3.77-3.74(m,0.8H),3.69-3.66(m,1H),3.62-3.52(m,3.6H),3.46-3.42(m,0.6H). 13C NMR(151MHz,D2O)δ109.2,96.3,92.2,83.81,83.79,81.3,80.4,77.1,76.58,76.55 ,75.0,71.0,70.3,69.2,68.6,67.4,61.2,61.1,60.9.HR-ESI-MS(m / z):calcdforC 11 H 20 O 10 Na + (M+Na) + :335.0949,found:335.0940.

[0109] Compound 46: Glycosyl donor 45 (188 mg, 0.31 mmol) and acceptor 30 (192 mg, 0.39 mmol) were glycosylated according to general method B to afford compound 46 (161 mg, 0.156 mmol, 50%). 1 H NMR (500MHz, CDCl3) δ8.18-8.12(m,2H,Ar),7.47(t,J=7.4Hz,1H,Ar),7.40-7.25( m,17H,Ar),7.21(ddd,J=17.3,7.4,3.0Hz,11H,Ar),7.11-7.07(m,2H,Ar),6.86-6 .79(m,2H,Ar),5.68(dd,J=10.2,3.9Hz,1H,D-H2),5.26(dd,J=13.6,3.9Hz,1H,D- H1),4.98(dd,J=11.1,7.4Hz,2H,Ar-CH2),4.85(dd,J=11.8,5.3Hz,2H,Ar-CH2),4 .76(d,J=7.5Hz,1H,A-H1),4.69(d,J=12.0Hz,1H,Ar-CH2),4.63-4.51(m,5H,Ar-C H2,D-H4,A-H4),4.24(d,J=1.6Hz,2H,Ar-CH2),4.17-4.10(m,2H,D-H3,A-H5),3.9 5-3.83(m,2H,A-H2,DH),3.77(s,3H,COOMe-CH3),3.62(t,J=8.5Hz,1H,DH),3.49( dd,J=9.8,2.9Hz,1H,A-H3),3.41(dd,J=8.6,5.2Hz,1H,DH),3.25(s,3H,MP-CH3). 13C NMR (126MHz, CDCl3) δ167.8,166.3,155.6,151.7,138.7,138.5,138.1,138.0,132.7,130.4,128.5,128.2,128.1,128.0,127.8,127.4,119. 4,114.5,103.4,97.1,79.7,78.4,77.3,76.2,75.4,74.8,74.2,73.4,72.9,72.7,72.2,71.9,70.6,68.3,55.7,52.2.HR-ESI-MS(m / z):calcd for C 62 H 62 O 13 Na + (M+Na) + :1053.4032,found:1053.42.

[0110] Compound 47: Compound 46 (150 mg, 0.15 mmol) was subjected to the general method for hydrolysis of 4-methoxyphenyl to obtain the intermediate compound (81 mg, 0.10 mmol, 67%). The intermediate compound (81 mg, 0.10 mmol) was subjected to the general method for acyl removal to obtain the intermediate compound (40 mg, 0.05 mmol, 50%). The intermediate compound (20 mg, 0.025 mmol) was subjected to the general method for catalytic hydrogenation to obtain compound 47 (8 mg, 0.022 mmol, 90%). 1 HNMR(600MHz,D2O)δ5.08(s,1H),4.79(s,1H),4.42(dd,J=21.3,7.6Hz,1H),4.20-4.15(m,2H),3.85(d, J=6.3Hz,1H),3.73(d,J=3.1Hz,1H),3.33-3.27(m,2H),3.08-3.01(m,1H),1.10(dd,J=15.0,6.3Hz,3H). 13 CNMR(151MHz,D2O)δ175.8,99.5,99.4,98.3,96.2,82.2,78.9,78.8,78.1,73.3,73.1,71.8,70.6,69.3, 69.3,69.2,69.1,68.9,68.7,68.6,68.6,68.0,62.1,61.1,60.2,57.7,57.5.HR-ESI-MS(m / z):calcdforC 12 H 20 O 12 Na +(M+Na) + :379.0847,found:379.2656.

[0111] Compound 48: Compound 38 (557 mg, 1.28 mmol) was glycosylated using General Method A to give Compound 48 (843 mg, 0.93 mmol, 73%). 1 HNMR(400MHz, CDCl3)δ7.89-7.70(m,5H,Ar),7.51-7.19(m,21H,Ar),5.92-5.84(m,1H, All-CH),5.84-5.81(m,1H,D-H4),5.29-5.22(m,1H,All-CH2),5.18(dd,J=10.3,1.7Hz, 1H,All-CH2),5.05-5.01(m,2H,D-H1,Ar-CH2),4.84(d,J=2.0Hz,1H,D-H2),4.80(dd,J= 5.8,2.3Hz,2H,A-H1,Ar-CH2),4.72(s,1H,Ar-CH2),4.69(s,1H,Ar-CH2),4.63(s,1H,Ar -CH2),4.60(s,1H,Ar-CH2),4.50(d,J=11.1Hz,1H,Ar-CH2),4.18-4.12(m,1H,All-CH2) ,4.09(t,J=2.5Hz,1H,All-CH2),4.02(dd,J=10.1,3.4Hz,1H,A-H2),3.97-3.90(m,2H,A r-CH2,A-H3),3.85-3.73(m,3H,D-H5,D-H3,A-H5),3.54(s,3H,Me),3.50(d,J=9.5Hz,1H ,A-H4),2.71-2.55(m,4H,Lev-CH2),2.13(s,2H,Lev-CH3),1.38(d,J=6.2Hz,3H,A-H6). 13CNMR(101MHz,CDCl3)δ206.0,171.5,168.2,138.6,138.2,137.9,135.9,133.8,133.3,133.0,1 28.3,128.3,128.2,128.2,128.1,128.1,128.0,127.9,127.8,127.7,127.58,127.57,127.5,1 27.4,127.2,126.7,126.1,126.1,125.9,117.3,97.6,96.4,79.9,79.2,77.4,77.3,77.1,76.7,75.2,74.9,74.4,72.7,72.2,72.1,71.8,69.4,69.0,68.3,67.8,52.4,38.0,29.7,28.0,18.1.

[0112] Compound 49: Compound 48 (228 mg, 0.25 mmol) was dissolved in pyridine / acetic acid (3.2 mL / 0.8 mL, 4 / 1), and 80% hydrazine hydrate was added under ice-bath, followed by stirring for 3 h. After completion of the reaction, the mixture was transferred to EtOAc and extracted with 1 M HCl and saturated sodium bicarbonate solution, respectively. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (EA / EA system, EA% = 30%) to give compound 49 (191 mg, 96%). 1HNMR (500MHz, CDCl3) δ7.84-7.75(m,4H,Ar),7.45-7.25(m,22H,Ar),5.95-5.81(m,1H,Al l-CH),5.25(ddd,J=17.2,10.5,2.0Hz,1H,All-CH2),5.20-5.14(m,1H,All-CH2),5.05(dd ,J=11.1,2.9Hz,1H,Ar-CH2),5.00(t,J=4.4Hz,1H,D-H1),4.91(d,J=1.7Hz,1H,A-H1),4. 81(dd,J=11.1,1.8Hz,2H,Ar-CH2),4.77(d,J=9.7Hz,1H,D-H2),4.73-4.69(m,4H,A-H2,Ar -CH2),4.65-4.58(m,3H,Ar-CH2),4.39(dd,J=3.4,1.8Hz,1H,D-H4),4.14(ddt,J=13.2,5 .1,1.7Hz,1H,All-CH2),4.11-4.04(m,1H,All-CH2),3.94(dtdd,J=28.2,9.7,6.6,3.1Hz, 5H,D-H5,D-H6,A-H3,D-H3),3.73(ddd,J=26.3,9.3,7.0Hz,1H,A-H5),3.59(s,3H,Me),3. 53(td,J=9.7,2.1Hz,1H,A-H4),1.38(dd,J=6.2,1.6Hz,3H,A-H6).HR-ESI-MS(m / z):calcd for C 48 H 52 O 11 K + (M+K) + :843.3140,found:843.3715.

[0113] Compound 50: Compound 49 (45 mg, 0.056 mmol) was dissolved in 2 mL of CH3COOH, and Pd(PPh3)4 (19 mg, 0.017 mmol) was added. The mixture was reacted at 80°C for 1 h. After the reaction was completed, the system was transferred to DCM, solid NaHCO3 was added to neutralize the acetic acid, and the mixture was extracted with distilled water and saturated NaCl solution in sequence. The organic phase was dried over anhydrous sodium sulfate, concentrated and the solvent was dried, and purified by silica gel column chromatography (DCM / MeOH = 10 / 1) to obtain compound 50 (33 mg, 0.043 mmol, 77%).

[0114] Compound 51: Compound 50 (34 mg, 0.044 mmol) was subjected to a general method of catalytic hydrogenation to obtain compound 51 (12 mg, 0.034 mmol, 77%). 1 HNMR (600MHz, D2O) δ5.06 (dd, J=4.3, 2.7Hz, 1H), 4.95 (d, J=3.9Hz, 0.7H, AH-1β), 4.91 (dd,J=5.9,1.6Hz,1H),4.79(s,0.3H,AH-1α),4.20(td,J=4.1,3.4,1.5Hz,1H),3.90-3 .82(m,1.3H),3.78(dd,J=3.6,1.7Hz,0.7H),3.76-3.67(m,2.3H),3.65(s,3H),3.56-3 .47(m,0.3H),3.33-3.24(m,1H),3.19(d,J=9.6Hz,0.3H),1.14(dd,J=8.8,6.3Hz,3H). 13 C NMR(151MHz,D2O)δ171.4,101.1,98.1,93.8,91.6,80.2,77.3,72.3,71.9,71.7, 71.1,70.1,69.1,68.7,68.6,68.2,67.7,52.8,16.7.HR-ESI-MS(m / z):calcdforC 13 H 21 O 11 Na + (M+Na) + :377.1054,found:377.1034.

[0115] Compound 53: Compound 50 (50 mg, 0.065 mmol) was dissolved in a mixed solvent of MeOH / H2O (1 mL / 1 mL, 1 / 1, v / v), and LiOH monohydrate (0.1 M / L, 27 mg, 0.65 mmol) was added. The mixture was allowed to react at room temperature for 5 h. After completion of the reaction, the pH was adjusted to slightly acidic using a hydrogen ion exchange resin. The mixture was filtered, the solvent was concentrated and dried, and the mixture was purified by silica gel column chromatography (DCM / MeOH = 10 / 1) to obtain Compound 52 (26 mg, 0.035 mmol, 54%). Compound 52 (26 mg, 0.035 mmol) was subjected to a general catalytic hydrogenation method to obtain Compound 53 (12 mg, 0.035 mmol, quant.). 1HNMR(600MHz,D2O)δ5.08(s,2H),4.79(s,1H),4.42(dd,J=21.3,7.6Hz,1H),4.22-4.12(m,5H),4.08-3.97(m,2H),3.89-3.79 (m,6H),3.72(ddt,J=21.8,15.6,7.1Hz,11H),3.61-3.45(m,3H),3.37-3.23(m,5H),3.09-3.00(m,2H),1.13(d,J=6.3Hz,9H). 13 C NMR(151MHz,D2O)δ101.2,93.9,92.8,91.5,81.5,80.9,79.9,76.5,75.0,72.6,7 2.5,71.2,70.7,70.4,68.9,68.5,,68.3,66.9,16.7.HR-ESI-MS(m / z):calcdforC 12 H 20 O 11 Na + (M+Na) + :363.0898,found:363.2270.

[0116] Experimental Example 5

[0117] The synthesis routes of Lycium barbarum oligosaccharides 55, 57, 59 and 61 are as follows:

[0118]

[0119] Compound 54: Glycosyl donor 39 (478.2 mg, 0.79 mmol) and acceptor 29 (85 mg, 0.166

[0120] mmol) were glycosylated according to the general method A to give compound 54 (62 mg, 0.067 mmol, 40%). 1H NMR (500MHz, CDCl3) δ8.11-8.03(m,2H,Ar),8.00-7.92(m,3H,Ar),7.57(td,J=7.7,3.0Hz,2 H,Ar),7.49-7.34(m,8H,Ar),7.30-7.25(m,2H,Ar),7.25-7.15(m,12H,Ar),6.61(s,1H,A-H 1),5.69(d,J=1.4Hz,1H,A-H2),5.36(d,J=1.4Hz,1H,D-H2),5.28(s,1H,D-H1),4.86(d,J=1 1.7Hz,1H,Ar-CH2),4.68(d,J=11.8Hz,1H,Ar-CH2),4.64-4.58(m,2H,D-H3,Ar-CH2),4.52(d ,J=12.1Hz,1H,Ar-CH2),4.45(dd,J=15.4,12.1Hz,2H,Ar-CH2),4.33(d,J=4.8Hz,1H,A-H3) ,4.23(q,J=4.8Hz,1H,D-H4),4.00(dd,J=11.3,4.2Hz,1H,D-H5),3.94(d,J=5.4Hz,1H,A-H4 ),3.77(dd,J=11.3,4.9Hz,1H,D-H5),3.59(dd,J=10.8,3.7Hz,1H,A-H5),3.53(dd,J=10.8, 5.0Hz,1H,A-H5),1.51(tt,J=8.0,5.2Hz,1H,ABz-CH),0.85(tt,J=8.2,2.8Hz,4H,ABz-CH2).

[0121] Compound 55: Compound 54 (23 mg, 0.025 mmol) and compound 36 (13.5 mg, 0.03 mmol) were azeotroped twice with toluene, dissolved in dry DCM, and added MS, and then catalyst PPh3AuOTf (9 mg, 0.015 mmol) was added and stirred at room temperature for 16 h. After the reaction was completed by TLC, triethylamine was added to quench the reaction, and the reaction was extracted with DCM / saturated NaHCO3 solution. The organic phase was dried over anhydrous sodium sulfate, concentrated and dried, and purified by column chromatography (PE / EA=5 / 1) to give compound 55 (9 mg, 0.075 mmol, 30%). 1H NMR (500MHz, CDCl3) δ8.02(t,J=7.6Hz,4H),7.95(d,J=7.7Hz,2H),7.55(dt,J=14.1,7.5Hz,3H),7.45-7.32(m,10H),7.22(ddt,J=28.5,20.9,6.3H z,20H),7.07(d,J=7.9Hz,2H),5.62(s,1H),5.57(s,1H),5.38(d,J=24.3H z,2H),5.25(s,2H),4.78(d,J=11.9Hz,1H),4.72-4.35(m,7H),4.27(d,J= 5.6Hz,1H),4.20(q,J=4.4Hz,1H),4.15(q,J=4.8Hz,1H),4.11(d,J=5.6Hz,1H),3.96(dd,J=11.5,4.1Hz,1H),3.91(d,J=5.5Hz,1H),3.86(dd,J=11 .3,4.0Hz,1H),3.76(dd,J=11.4,4.1Hz,1H),3.68(dd,J=11.4,3.8Hz,1H),3.56(dd,J=10.8,3.8Hz,1H),3.49(dd,J=10.7,5.0Hz,1H),2.28(s,2H).

[0122] Compound 57: Compound 45 (27 mg, 0.042 mmol) was dissolved in DCM (1 mL). 4AMS molecular sieves were added, followed by BSP (10.55 mg, 0.048 mmol) and TTBP (15.65 mg, 0.063 mmol). The mixture was placed in a -60°C reaction vessel. Tf2O (9 μL, 0.052 mmol) was added, and the mixture was allowed to react at -60°C for 30 min. After TLC analysis of the complete reaction, 1-octene (8.5 μL, 0.052 mmol) was added, and the reaction was continued at -60°C for 15 min. The temperature was then lowered to -78°C, and compound 56 (27 mg, 0.053 mmol) dissolved in 1 mL of DCM was added. The reaction was continued at -78°C for 3 h. After TLC analysis of the complete reaction, triethyl phosphite (22 μL, 0.126 mmol) was added, and the reaction was continued at -78°C for 1 h before returning to room temperature. The reaction solution was extracted with DCM / saturated NaHCO3 solution and saturated NaCl solution. The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (PE / EA=5 / 1, v / v) to give compound 13 (17 mg, 0.015 mmol, 37%). 1HNMR (400MHz, CDCl3) δ7.97 (dd, J=7.6, 5.0Hz, 3H, Ar), 7.74 (d, J=7.6Hz, 1H, Ar), 7.64-7.51 (m, 4H, Ar), 7.47-7.05 (m, 22H, Ar), 5.70-5 .61(m,1H,D-H1),5.56(t,J=9.7Hz,1H,A-H2),5.04(d,J=11.7Hz,1H,Ar-CH2),4.85(dd,J=29.1,12.1Hz,2H),4.73-4.57(m,3H),4.52( d,J=7.9Hz,1H),4.43(d,J=6.8Hz,2H),4.29(t,J=10.6Hz,2H),4.10(s,1H),3.92(dt,J=15.3,4.5Hz,2H),3.73-3.64(m,2H),3.64-3.5 7(m,1H),3.55(dd,J=8.2,4.6Hz,1H),3.51-3.42(m,2H),2.48(ddq,J=27.2,14.0,7.0Hz,2H,SEt-CH2),1.01(t,J=7.5Hz,3H,SEt-CH3).

[0123] Compound 59: Compound 35 (106 mg, 0.20 mmol) was glycosylated using General Method B to afford compound 59 (85 mg, 0.088 mmol, 49%). 1 HNMR (500MHz, CDCl3) δ7.86-7.52(m,3H,Ar),7.45-7.05(m,34H,Ar),5.31(t,J=2.5Hz,1H,D-H 2),4.99(d,J=11.1Hz,1H),4.90(d,J=11.4Hz,1H),4.85-4.79(m,3H),4.72-4.57(m,7H),4.53- 4.38(m,5H),3.98-3.89(m,2H),3.85-3.78(m,2H),3.75(s,1H),3.57(dt,J=12.5,6.3Hz,1H), 3.47-3.35(m,3H,Ac),2.09(s,3H,STol-CH3),1.19(d,J=5.9Hz,3H,A-H6).ESI-MS(m / z):calcd for C 60 H 62 O 11 Na + (M+Na) + :981.4184,found:981.42.

[0124] Compound 60: Glycosyl donor 32 (212 mg, 0.32 mmol) and acceptor 34 (67 mg, 0.134 mmol) were glycosylated using General Method A to give compound 60 (85 mg, 0.09 mmol, 67%, α / β = 2.3 / 1). 1 H NMR 81α(400MHz, CDCl3)δ7.87-7.77(m,4H,Ar),7.48(ddd,J=9.6,6.9,2.1Hz,3H,Ar),7.36-7.2 8(m,13H,Ar),7.22-7.09(m,5H,Ar),6.90(d,J=8.1Hz,1H,Ar),5.83(dd,J=3.5,1.7Hz,1H,D- H4),5.35(d,J=1.7Hz,1H,A-H2),5.07(d,J=11.2Hz,1H,Ar-CH2),4.98(dd,J=17.3,2.6Hz,2H ,D-H5,Ar-CH2),4.83(dd,J=16.0,11.5Hz,2H,Ar-CH2),4.71(d,J=11.8Hz,2H,Ar-CH2),4.64 (d,J=12.0Hz,1H,Ar-CH2),4.52(dd,J=15.7,11.5Hz,2H,Ar-CH2),4.32-4.29(m,1H,A-H1),4 .21(dd,J=9.4,6.1Hz,1H,A-H5),4.00(dd,J=10.0,3.4Hz,1H,D-H3),3.88(dd,J=9.5,3.0Hz, 1H,A-H3),3.79(dd,J=10.1,3.5Hz,1H,D-H2),3.59(s,3H,Me),3.51(s,1H,A-H4),2.70-2.53 (m,4H,Lev-CH2),2.36(s,3H,Lev-CH3),2.14(s,3H,STol-CH3),1.41(d,J=6.2Hz,3H,A-H6). 13 C NMR (101MHz, CDCl3) δ206.0,171.5,168.2,138.0,137.5,132.1,129.8,128.5,127.3,126.7,126.3,125.8,96.9,85 .7,80.0,79.3,77.3,76.1,75.2,74.8,74.5,72.9,72.1,71.8,69.4,68.9,52.5,38.0,29.8,28.0,23.4,21.1,18.0.

[0125] Compound 61: Glycosyl donor 60 (23 mg, 0.024 mmol) and acceptor 58 (14 mg, 0.0264 mmol) were glycosylated according to general method B to give compound 16 (8 mg, 0.088 mmol, 24%). 1 H NMR(500MHz, CDCl3)δ7.74(dt,J=11.1,5.7Hz,3H),7.46-7.06(m,39H),5.73(dt,J=15.4,3.0Hz,1H),5.5 7(d,J=3.7Hz,1H),5.05(d,J=1.7Hz,1H),4.91(dd,J=19.5,11.6Hz,2H),4.81(d,J=11.5Hz,1H),4.76-4. 36(m,13H),4.23(d,J=12.8Hz,1H),4.04(d,J=12.7Hz,1H),3.96-3.84(m,4H),3.79-3.64(m,5H),3.50(s ,2H),3.42-3.24(m,3H),3.20-3.10(m,1H),2.64-2.44(m,4H),2.09-2.01(m,3H),1.33(d,J=6.1Hz,3H).

[0126] Application Example 1 Anti-pancreatic cancer activity test of black wolfberry oligosaccharide and its derivatives

[0127] Specific test operations and steps:

[0128] (1) MTT assay to detect the effects of Lycium barbarum oligosaccharides and their derivatives on the proliferation of pancreatic cancer cells PANC-1, BxPC-3, and AsPC-1

[0129] Pancreatic cancer cells PANC-1, BxPC-3, and AsPC-1 (cell density of 1×10 4The suspension was inoculated into a 96-well plate. 100 μL was added to each well, and three replicates were set up. A blank group (equal volume of culture medium) and a control group (cells incubated with equal volume of culture medium) were also set up and cultured overnight. Different concentrations of aqueous solutions of black wolfberry oligosaccharides and their derivatives 41, 44, 47, 51 and 53 were added to the 96-well plate, respectively, and cultured together with the blank group and the control group at 37°C and 5% CO2 for 72 hours. 10 μL of thiazolyl blue (MTT) solution (5 mg / mL) was added and cultured in an incubator for another 4 hours. The culture medium was discarded, and 150 μL of dimethyl sulfoxide was added to dissolve the formazan in the cells. The 96-well plate was shaken for 15-30 minutes, and the absorbance at 490 nm was detected using a microplate reader. The cell viability was calculated according to the following formula: Cell viability = (OD value of the experimental group - OD value of the blank group) / (OD value of the control group - OD value of the blank group) × 100%.

[0130] Five synthetic oligosaccharide fragments and the positive control drug gemcitabine were used to treat pancreatic cancer cells at different concentrations. The results are as follows Figure 1 、 2 As shown in Figures 3 and 4, compounds 44 and 53 showed a certain inhibitory effect on pancreatic cancer cells PANC-1, AsPC-1, and BxPC-3. Among them, the inhibition rates of compounds 44 and 53 on gemcitabine-resistant PANC-1 cells reached 78.3% and 57.84%, and the highest inhibition rates on AsPC-1 and BxPC-3 cells reached 45.4% and 47.5%, respectively. The most active compound 44 was selected for IC 50 The test results are as follows Figure 4 As shown, the IC of compound 44 against pancreatic cancer cells PANC-1 50 The value was 263.5 μM. Therefore, α-L-arabinose-(1→3)-L-galactose and α-L-rhamnose-(1→4)-α-D-galacturonic acid may be the active structural domains of Lycium ruthenicum polysaccharide, and subsequent synthesis of larger oligosaccharides containing this structural fragment can be continued.

[0131] In summary, it can be seen from the examples that Lycium barbarum oligosaccharides can become a potential carbohydrate drug for the treatment of pancreatic cancer.

Claims

1. A lycium barbarum oligosaccharide derivative, characterized by: The derivative is a compound represented by formula (3a) or a pharmaceutically acceptable salt thereof:

2. The lycium ruthenicum oligosaccharide derivative according to claim 1, characterized in that: The pharmaceutically acceptable salt is any one of sodium salt, potassium salt, calcium salt, magnesium salt, zinc salt and meglumine salt.

3. The method for preparing the ruthenic wolfberry oligosaccharide derivative according to claim 1, wherein: The method comprises the following steps: glycosidation reaction of glycosyl donor 9 and glycosyl acceptor 12 to obtain disaccharide 3; removal of acyl protecting group of disaccharide 3 under alkaline conditions, and removal of aromatic protecting group by reduction reaction to complete deprotection and obtain fully deprotected disaccharide 3a. The synthetic route is shown below: Wherein, LG is a leaving group of the glycosyl donor and is any one of halogen, trichloroacetimidyloxy, N-phenyltrifluoroacetimidyloxy, dibenzylphosphoxy, o-alkynylbenzoyloxy, o-methoxycarbonylalkynylphenylthio, methylthio, ethylthio, phenylthio, and p-tolylthio; PG5 is acetoxy, benzoyloxy, pivaloyloxy, chloroacetoxy, levulinyloxy, allyloxycarbonyloxy, methoxy, benzyloxy, 2-naphthylmethoxy, p-methoxybenzyloxy, allyloxy, p-methoxyphenoxy, trityloxy, monomethoxytrityloxy, dimethoxytrityloxy, tert-butyldimethylsilyl PG2 and PG6 are any one of acetyl, benzoyl, pivaloyl, chloroacetyl, levulinyl and allyloxycarbonyl; PG3, PG4, PG7 and PG8 are any one of acetyl, benzoyl, pivaloyl, chloroacetyl, levulinyl, allyloxycarbonyl, methyl, benzyl, 2-naphthylmethyl, p-methoxybenzyl, allyl, p-methoxyphenyl, trityl, monomethoxytrityl, dimethoxytrityl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl and triethylsilyl.

4. The method for preparing the ruthenic wolfberry oligosaccharide derivative according to claim 3, wherein: The accelerator is any one of boron trifluoride etherate, trifluoromethanesulfonic acid, trimethylsilyl trifluoromethanesulfonate, silver carbonate, N-iodosuccinimide, trifluoromethanesulfonic acid, triphenylphosphine gold trifluoromethanesulfonate, and triphenylphosphine gold bis(fluorosulfonyl)imide.

5. The method for preparing the ruthenic wolfberry oligosaccharide derivative according to claim 3, wherein: The conditions for the reduction reaction to remove the aromatic protecting group are: a mixed solvent consisting of dichloromethane or tetrahydrofuran or ethyl acetate and tert-butanol or methanol, water, and acetic acid, a 10% palladium carbon catalyst or palladium hydroxide, and hydrogen, and stirring at room temperature for 48 hours; or sodium, liquid ammonia, tetrahydrofuran, and tert-butanol, and stirring at -78°C for 30 minutes.

6. The method for preparing the ruthenic wolfberry oligosaccharide derivative according to claim 3, wherein: The molar ratio of the glycosyl donor to the glycosyl acceptor in the synthetic route is (1-2):1; the glycosidation reaction conditions are as follows: dissolving the glycosyl donor and the glycosyl acceptor in a dry solvent, adding molecular sieves and a promoter, and stirring the reaction at a set temperature for 3-7 hours to obtain the target oligosaccharide fragment; the dry solvent is at least one of anhydrous dichloromethane, anhydrous ether, anhydrous toluene, anhydrous methanol, anhydrous tetrahydrofuran, anhydrous acetonitrile, anhydrous N,N-dimethylformamide, and anhydrous dioxane; and the stirring at the set temperature is stirring the reaction at room temperature of 25°C, stirring the reaction in an ice-water mixture of 0°C, stirring the reaction in a mixture of ice and sodium chloride at -5 to -20°C, stirring the reaction in a mixture of acetonitrile and dry ice at -40°C, stirring the reaction in a mixture of acetone and dry ice at -60°C, or stirring the reaction in a mixture of acetone and dry ice at -78°C.

7. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the lycium ruthenicum oligosaccharide derivative according to any one of claims 1 to 2, a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

8. Use of the pharmaceutical composition according to claim 7 in the preparation of a drug for preventing or treating pancreatic cancer.

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