Lycium ruthenicum oligosaccharide derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of chemical synthesis, and particularly relates to a lycium ruthenicum oligosaccharide derivative as well as a preparation method and application thereof, efficient construction of 1, 2-cis-alpha-galacturonic acid glucosidic bonds and other glucosidic bonds in an oligosaccharide structure is realized by utilizing an acyl remote participation effect, a steric hindrance effect, a solvent effect, an end group position effect and the like, and the preparation method and application of the lycium ruthenicum oligosaccharide derivative are suitable for industrial production. The first-time chemical synthesis of the lycium ruthenicum oligosaccharide and the derivative thereof is realized; the lycium ruthenicum oligosaccharide fragment provided by the invention has an inhibiting effect on pancreatic cancer cells, and provides a theoretical basis for defining the active fragment (active structural domain) of lycium ruthenicum polysaccharide, carrying out deeper functional mechanism research and developing a new pancreatic cancer resisting drug based on the active fragment; the compound has a good application prospect in preparation of novel anti-cancer drugs.

Description

Technical Field

[0001] The present invention belongs to the technical field of chemical synthesis, and particularly relates to a black wolfberry oligosaccharide derivative, a preparation method thereof, and an application thereof. Background Art

[0002] Pancreatic cancer is one of the most lethal malignant tumors. Compared with other malignant tumors, pancreatic cancer has a high degree of malignancy, insidious onset, lack of typical symptoms, high invasiveness, may have tissue infiltration and distant metastasis in the early stage, low surgical resection rate, high recurrence rate, and poor prognosis. The five-year survival rate of patients is only 8%. For patients with locally advanced or metastatic pancreatic cancer, the five-year survival rate is only 2%. At present, the treatment methods for pancreatic cancer include surgery, embolization, radiotherapy, chemotherapy, and immunotherapy, etc. Surgery is the only possible radical treatment method, but only 20% of patients have the opportunity for surgery. Even for these patients who have undergone radical surgical treatment, the probability of postoperative recurrence and metastasis is still very high. Chemotherapy is one of the main treatment methods for pancreatic cancer. Its main drug is gemcitabine, or it is administered in combination with other drugs, such as 5-fluorouracil, etc. However, during the treatment process, patients are prone to develop drug resistance to chemotherapy drugs. Therefore, the treatment methods for pancreatic cancer are extremely limited, the mortality rate remains high, and there is an urgent need to develop new treatment regimens and drugs.

[0003] Traditional Chinese medicine polysaccharides have significant anti-tumor activities and low toxicity to the human body, showing great application potential. Research has found that polysaccharides extracted from natural sources such as Lycium ruthenicum polysaccharide, Polygonatum cyrtonema Hua polysaccharide, Carthamus tinctorius polysaccharide, Polygala tenuifolia polysaccharide, Zea mays polysaccharide, Ophiopogon japonicus polysaccharide, Lonicera japonica polysaccharide, and Gastrodia elata polysaccharide have good anti-pancreatic cancer activities and are important lead compounds for the development of new anti-pancreatic cancer drugs. Lycium ruthenicum Murr is a perennial shrub of the genus Lycium in the Solanaceae family, containing various active ingredients such as polysaccharides, flavonoids, anthocyanins, phenolic acids, and essential oils. In previous studies by the research group, two homogeneous polysaccharides, LRP1-S2 and LRP3-S1, with relative molecular weights of 17.0 kDa and 114.8 kDa respectively, were extracted and isolated from Lycium ruthenicum, and the structures of the two polysaccharides were characterized. In vitro activity experiments showed that both LRP1-S2 and LRP3-S1 could inhibit the proliferation of pancreatic cancer cells BxPC-3, AsPC-1, and PANC-1, and had no obvious cytotoxicity to normal pancreatic cells HPDE6-C7 and liver cells LO2. In addition, LRP1-S2 also had a good inhibitory effect on the growth of solid tumors in the BxPC-3 nude mouse xenograft model of pancreatic cancer, and the inhibitory effect was better than that of the positive control group of gemcitabine. Further mechanism studies showed that LRP1-S2 might induce apoptosis of BxPC-3 cells by blocking the p38 MAPK / NF-κB and GSK-3β / β-Catenin signaling pathways; LRP3-S1 could weaken the invasion ability of pancreatic cancer cells BxPC-3, probably due to the polysaccharide blocking the p38 MAPK signaling pathway and the FAK / AKT / GSK-3β signaling pathway.

[0004] The two polysaccharides can block the same signaling pathways, 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). The structures of these two polysaccharides are unique and are important lead compounds for the development of new anti-pancreatic cancer carbohydrate drugs. However, their active fragments (active determinants) are not yet clear. In addition, the discovery of the anti-pancreatic cancer target molecules of the two polysaccharides and the related mechanism studies are not deep enough and systematic enough, and are still in the initial stage. It is very challenging to obtain polysaccharides with clear structures and homogeneity through natural extraction methods. The quality of polysaccharides is affected by many factors such as climate, cultivation conditions, and extraction methods; the molecular weight of polysaccharides is relatively large and the solubility is also relatively poor, which also affects its pharmacological activity in the body; the purification efficiency is low, the structure determination is difficult, and it is difficult to obtain sufficient amounts of oligosaccharide fragments with clear structures, which limits its application in the development of carbohydrate drugs. Therefore, it is crucial to prepare oligosaccharide fragments with clear structures and homogeneity through chemical synthesis methods to explore the active fragments (active determinants) of the two polysaccharides for the research and production of innovative drugs based on Lycium ruthenicum polysaccharide against pancreatic cancer. Summary of the Invention

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

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

[0007] A Lycium ruthenicum oligosaccharide derivative, wherein the derivative is a compound of the structure shown in formula (1) to (8) or a pharmaceutically acceptable salt thereof:

[0008]

[0009] Wherein, PG 1 , PG 5 , PG 10 , PG 17 is any one of hydroxyl, acetoxy, benzoyloxy, pivaloyloxy, chloroacetoxy, acetylpropionyloxy, allyloxycarbonyloxy, methoxy, benzyloxy, 2-naphthalenemethoxy, p-methoxybenzyloxy, allyloxy, p-methoxyphenoxy, trityloxy, monomethoxytrityloxy, dimethoxytrityloxy, tert-butyldimethylsilyloxy, tert-butyldiphenylsilyloxy, triethylsilyloxy, halogen, trichloroacetimidate, N-phenyltrifluoroacetimidate, dibenzylphosphate, o-alkynylbenzoyloxy, o-methoxycarbonylalkynylphenylthio, methylthio, ethylthio, phenylthio, p-tolylthio; PG 2 , PG 6 , PG 11 , PG 14 can be any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, acetylpropionyl, allyloxycarbonyl; PG 3 , PG 4 , PG 7 , PG 8 , PG 9 , PG 12 , PG 15 , PG 16 is any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, acetylpropionyl, allyloxycarbonyl, methyl, benzyl, 2-naphthalenemethyl, p-methoxybenzyl, allyl, p-methoxyphenyl, trityl, monomethoxytrityl, dimethoxytrityl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, triethylsilyl, benzylidene acetal, isopropylidene ketal; PG 13 is any one of hydrogen, methyl, ethyl, tert-butyl, benzyl, allyl, trichloroethyl, trimethylsilylethyl; PG18 is any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, acetylpropionyl, 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 ruthenicum oligose derivative includes the following steps:

[0012] S1. Glycosylation reaction is carried out on glycosyl donor 9 and glycosyl acceptor 10 to obtain disaccharide 1; disaccharide 1 is deacylated under alkaline conditions, and the aromatic protecting group is removed by reduction reaction to complete deprotection, obtaining fully deprotected disaccharide 1a; the synthetic route is as follows:

[0013]

[0014] Among them, LG is the leaving group of the glycosyl donor, which is any one of halogen, trichloroacetimidate, N-phenyltrifluoroacetimidate, dibenzylphosphate, o-alkynylbenzoyloxy, o-methoxycarbonylalkynylphenylthio, methylthio, ethylthio, phenylthio, and p-tolylthio;

[0015] S2. Selectively remove the protecting group PG of disaccharide 1 4 to obtain disaccharide 11, and then carry out glycosylation reaction with glycosyl donor 9 under the catalysis of a promoter to obtain trisaccharide 2; trisaccharide 2 is deacylated under alkaline conditions, and the aromatic protecting group is removed by reduction reaction to complete deprotection, obtaining fully deprotected trisaccharide 2a; the synthetic route is as follows:

[0016]

[0017] S3. Carry out glycosylation reaction on glycosyl donor 9 and glycosyl acceptor 12 to obtain disaccharide 3; disaccharide 3 is deacylated under alkaline conditions, and the aromatic protecting group is removed by reduction reaction to complete deprotection, obtaining fully deprotected trisaccharide 3a; the synthetic route is as follows:

[0018]

[0019] S4. Carry out glycosylation reaction on glycosyl donor 13 and glycosyl acceptor 14 to obtain disaccharide 4; disaccharide 4 is deacylated under alkaline conditions, and the aromatic protecting group is removed by reduction reaction to complete deprotection, obtaining fully deprotected trisaccharide 4a; the synthetic route is as follows:

[0020]

[0021] S5. Glycosylation reaction is carried out on glycosyl donor 13 and glycosyl acceptor 15 to obtain disaccharide 5; the acyl protecting group of disaccharide 5 is removed under alkaline conditions, and the aromatic protecting group is removed through a reduction reaction to complete deprotection, obtaining fully deprotected trisaccharide 5a; the synthetic route is as follows:

[0022]

[0023] S6. Glycosylation reaction is carried out on glycosyl donor 16 and glycosyl acceptor 14 to obtain disaccharide 6; the acyl protecting group of disaccharide 6 is removed under alkaline conditions, and the aromatic protecting group is removed through a reduction reaction to complete deprotection, obtaining fully deprotected trisaccharide 6a; the synthetic route is as follows:

[0024]

[0025] S7. Glycosylation reaction is carried out on glycosyl donor 17 and glycosyl acceptor 18, and the remote participation effect of the acyl group at the C4 position is utilized to complete the construction of the 1,2-cis-glycosidic bond, obtaining disaccharide 7; the acyl protecting group of disaccharide 7 is removed under alkaline conditions, and the aromatic protecting group is removed through a reduction reaction to complete deprotection, obtaining fully deprotected disaccharide 7a; the synthetic route is as follows:

[0026]

[0027] S8. Glycosylation reaction is carried out on glycosyl donor 19 and glycosyl acceptor 14 to obtain trisaccharide 8; the acyl protecting group of trisaccharide 7 is removed under alkaline conditions, and the aromatic protecting group is removed through a reduction reaction to complete deprotection, obtaining fully deprotected trisaccharide 8a; the synthetic route is as follows:

[0028]

[0029] Furthermore, the promoter is any one of boron trifluoride diethyl ether, trifluoromethanesulfonic acid, trimethylsilyl trifluoromethanesulfonate, silver carbonate, trifluoromethanesulfonic acid, N-iodosuccinimide and trifluoromethanesulfonic acid, N-iodosuccinimide and N-iodosuccinimide and trifluoromethanesulfonic acid, triphenylphosphine gold trifluoromethanesulfonate, triphenylphosphine gold bisfluorosulfonimide.

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

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

[0032] A pharmaceutical composition, which comprises the black goji berry oligosaccharide derivative described above, its pharmaceutically acceptable salt, and a pharmaceutically acceptable excipient.

[0033] Use of a pharmaceutical composition in the preparation of a drug for preventing or treating pancreatic cancer.

[0034] The advantages of the present invention are as follows: the present invention utilizes the acyl remote participation effect, steric hindrance effect, solvent effect, and anomeric effect, etc. to achieve the efficient construction of 1,2-cis-α-galacturonic acid glycosidic bonds and other glycosidic bonds in the oligosaccharide structure, and realizes the first chemical synthesis of black goji berry oligosaccharides and their derivatives; the black goji berry oligosaccharide fragment provided by the present invention has an inhibitory effect on pancreatic cancer cells, provides a theoretical basis for clarifying the active fragment (active domain) of black goji berry polysaccharide, for conducting more in-depth functional mechanism research and the development of new anti-pancreatic cancer drugs based on its active fragment, and has good application prospects in the preparation of new anti-cancer drugs. Description of the Drawings

[0035] Figure 1 : Effects of synthesized black goji berry oligosaccharide fragments and gemcitabine on the growth of pancreatic cancer cell line PANC-1.

[0036] Figure 2 : Effects of synthesized black goji berry oligosaccharide fragments and gemcitabine on the growth of pancreatic cancer cell line AsPC-1.

[0037] Figure 3 : Effects of synthesized black goji berry oligosaccharide fragments and gemcitabine on the growth of pancreatic cancer cell line BxPC-3.

[0038] Figure 4 : IC of disaccharide 44 against pancreatic cancer cell line PANC-1 50 Test relationship diagram. Detailed Description of the Invention

[0039] Example

[0040] A Lycium ruthenicum Murr. oligosaccharide derivative, which is a compound having a structure shown in formula (1) to (8) or a pharmaceutically acceptable salt thereof:

[0041]

[0042] wherein, PG 1 , PG 5 , PG 10 , PG 17 is any one of hydroxyl, acetoxy, benzoyloxy, pivaloyloxy, chloroacetoxy, acetylpropionyloxy, allyloxycarbonyloxy, methoxy, benzyloxy, 2-naphthalenemethoxy, p-methoxybenzyloxy, allyloxy, p-methoxyphenoxy, trityloxy, monomethoxytrityloxy, dimethoxytrityloxy, tert-butyldimethylsilyloxy, tert-butyldiphenylsilyloxy, triethylsilyloxy, halogen, trichloroacetimidate, N-phenyltrifluoroacetimidate, dibenzylphosphate, o-alkynylbenzoyloxy, o-methoxycarbonylalkynylphenylthio, methylthio, ethylthio, phenylthio, p-tolylthio; PG 2 , PG 6 , PG 11 , PG 14 can be any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, acetylpropionyl, allyloxycarbonyl; PG 3 , PG 4 , PG 7 , PG 8 , PG 9 , PG 12 , PG 15 , PG 16 is any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, acetylpropionyl, allyloxycarbonyl, methyl, benzyl, 2-naphthalenemethyl, p-methoxybenzyl, allyl, p-methoxyphenyl, trityl, monomethoxytrityl, dimethoxytrityl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, triethylsilyl, benzylidene acetal, isopropylidene ketal; PG 13 is any one of hydrogen, methyl, ethyl, tert-butyl, benzyl, allyl, trichloroethyl, trimethylsilylethyl; PG 18 is any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, acetylpropionyl, 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 preparation method of a black goji berry oligosaccharide derivative, comprising the following steps:

[0045] S1. Glycosylation reaction is carried out on glycosyl donor 9 and glycosyl acceptor 10 to obtain disaccharide 1; disaccharide 1 removes the acyl protecting group under alkaline conditions and removes the aromatic protecting group through a reduction reaction to complete deprotection, obtaining fully deprotected disaccharide 1a; the synthetic route is as follows:

[0046]

[0047] Among them, LG is the leaving group of the glycosyl donor, which is any one of halogen, trichloroacetimidate, N-phenyltrifluoroacetimidate, dibenzyl phosphate, o-alkynylbenzoyl, o-methoxycarbonylalkynylphenylthio, methylthio, ethylthio, phenylthio, p-tolylthio;

[0048] S2. Selectively remove the protecting group PG of disaccharide 1 4 to obtain disaccharide 11, and then carry out glycosylation reaction with glycosyl donor 9 under the catalysis of a promoter to obtain trisaccharide 2; trisaccharide 2 removes the acyl protecting group under alkaline conditions and removes the aromatic protecting group through a reduction reaction to complete deprotection, obtaining fully deprotected trisaccharide 2a; the synthetic route is as follows:

[0049]

[0050] S3. Glycosylation reaction is carried out on glycosyl donor 9 and glycosyl acceptor 12 to obtain disaccharide 3; disaccharide 3 removes the acyl protecting group under alkaline conditions and removes the aromatic protecting group through a reduction reaction to complete deprotection, obtaining fully deprotected trisaccharide 3a; the synthetic route is as follows:

[0051]

[0052] S4. Glycosylation reaction is carried out on glycosyl donor 13 and glycosyl acceptor 14 to obtain disaccharide 4; disaccharide 4 removes the acyl protecting group under alkaline conditions and removes the aromatic protecting group through a reduction reaction to complete deprotection, obtaining fully deprotected trisaccharide 4a; the synthetic route is as follows:

[0053]

[0054] S5. Glycosylation reaction is carried out on glycosyl donor 13 and glycosyl acceptor 15 to obtain disaccharide 5; disaccharide 5 removes the acyl protecting group under alkaline conditions and removes the aromatic protecting group through a reduction reaction to complete deprotection, obtaining fully deprotected trisaccharide 5a; the synthetic route is as follows:

[0055]

[0056] S6. Glycosylation reaction is carried out on the glycosyl donor 16 and the glycosyl acceptor 14 to obtain the disaccharide 6; the acyl protecting group of the disaccharide 6 is removed under alkaline conditions, and the aromatic protecting group is removed through a reduction reaction to complete the deprotection, obtaining the fully deprotected trisaccharide 6a; the synthetic route is as follows:

[0057]

[0058] S7. Glycosylation reaction is carried out on the glycosyl donor 17 and the glycosyl acceptor 18. Utilizing the long-range participation effect of the acyl group at the C4 position, the construction of the 1,2-cis-glycosidic bond is completed to obtain the disaccharide 7; the acyl protecting group of the disaccharide 7 is removed under alkaline conditions, and the aromatic protecting group is removed through a reduction reaction to complete the deprotection, obtaining the fully deprotected disaccharide 7a; the synthetic route is as follows:

[0059]

[0060] S8. Glycosylation reaction is carried out on the glycosyl donor 19 and the glycosyl acceptor 14 to obtain the trisaccharide 8; the acyl protecting group of the trisaccharide 7 is removed under alkaline conditions, and the aromatic protecting group is removed through a reduction reaction to complete the deprotection, obtaining the fully deprotected trisaccharide 8a; the synthetic route is as follows:

[0061]

[0062] Furthermore, the promoter is any one of boron trifluoride diethyl ether, trifluoromethanesulfonic acid, trimethylsilyl trifluoromethanesulfonate, silver carbonate, trifluoromethanesulfonic acid, N-iodosuccinimide and trifluoromethanesulfonic acid, N-iodosuccinimide and N-iodosuccinimide and trifluoromethanesulfonic acid, triphenylphosphine gold trifluoromethanesulfonate, triphenylphosphine gold bisfluorosulfimide.

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

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

[0065] A pharmaceutical composition, which comprises the black goji berry oligosaccharide derivative as described above, its pharmaceutically acceptable salt, and a pharmaceutically acceptable excipient.

[0066] Use of a pharmaceutical composition in the preparation of a drug for preventing or treating pancreatic cancer.

[0067] Experimental examples

[0068] General experimental methods

[0069] The anhydrous solvents used in the experiments were commercial reagents (Energy Chemical). 1 1H NMR, 13 13C NMR, 1 1H- 1 1H- 1 1H- 13 1H COSY, 1H-13C HSQC were all measured by a Bruker AVANCE III 500 nuclear magnetic resonance spectrometer, with TMS as the internal standard, measured at 25 °C, and different peak patterns were represented by singlet (s), doublet (d), triplet (t), quartet (dd), multiplet (m), etc. The chemical shift (δ) was recorded in ppm, and the coupling constant (J) was recorded in Hz. The mass spectrometry was measured by a Thermo Scientific Ultimate 3000 Orbitrap Exploris 240 ultra-high performance liquid chromatography-high resolution mass spectrometry in the positive and negative ion full scan modes. Column chromatography and thin layer chromatography (TLC) used 200-300 mesh column chromatography silica gel and thin layer chromatography silica gel GF254 type silica gel plates produced by Qingdao Marine Chemical Factory, Shandong. The color development method for experimental detection used a 5% (v / v) sulfuric acid-ethanol solution to develop color under a hot air gun and ultraviolet light.

[0070] General glycosylation method A: Combine the glycosyl donor (1.5 equivalents) and the glycosyl acceptor (1.0 equivalent), dissolve in toluene, and evaporate to dryness to remove water twice (temperature 30 °C). Under nitrogen protection, dissolve the above raw materials in anhydrous dichloromethane (reaction concentration 0.1 - 0.5 mol / L), add molecular sieve, stir at -45 °C for 10 min, add TMSOTf (0.3 equivalent), and stir and react at -45 °C for 2 h. After monitoring the reaction by TLC until completion, transfer the reaction solution with dichloromethane, extract with dichloromethane and saturated sodium bicarbonate, dry the organic phase with anhydrous sodium sulfate, evaporate to dryness, and purify by medium-pressure semi-preparative chromatography column to obtain the target oligosaccharide.

[0071] General glycosylation method B: Combine the glycosyl donor (1.5 equivalents) and the glycosyl acceptor (1.0 equivalent), and azeotrope with toluene twice. Under nitrogen protection, dissolve in 5 mL of DCM, add molecular sieve, NIS (2.0 equivalents), add TMSOTf (0.2 equivalent) at 0 °C, and stir and react at 0 °C for 5 h. After monitoring the reaction by TLC until completion, transfer the reaction solution with DCM, filter, extract with DCM / saturated sodium thiosulfate and saturated NaHCO 3 solution extraction, dry the organic phase with anhydrous sodium sulfate, concentrate and evaporate to dryness, and purify by column chromatography to obtain the target oligosaccharide.

[0072] General method for the hydrolysis of 4-methoxyphenyl: Dissolve the compound (1.0 equivalent) in DCM / MeCN / H 2 O (reaction concentration 0.1 - 0.5 mol / L, 2 / 1 / 2, v / v / v), add CAN (4.0 equivalents), and react at room temperature for 5 h. After monitoring the reaction of the raw materials by TLC until complete, transfer the reaction solution with DCM, extract with DCM / 1M hydrochloric acid and saturated sodium bicarbonate solution, dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure, and purify by medium-pressure semi-preparative chromatography to obtain the target compound.

[0073] General method for the removal of silyl ether protecting groups: Dissolve compound 25 (1.0 equivalent) in tetrahydrofuran (reaction concentration 0.1 - 0.5 mol / L), add acetic acid (1.5 equivalents) and TBAF (1.5 equivalents) at 0 °C, and react in an ice bath for 12 h. After detecting the reaction of the raw materials by TLC until complete, add dichloromethane to dilute the reaction solution, extract with dichloromethane and saturated sodium bicarbonate solution and saturated brine, dry the organic layer with anhydrous sodium sulfate, concentrate under reduced pressure, and purify by silica gel column chromatography to obtain the target compound.

[0074] General method for thioglycoside hydrolysis: Dissolve the compound (1.0 equiv) in acetone / water (reaction concentration 0.1 mol / L, 4 / 1, v / v), add trichloroisocyanuric acid (1.7 equiv), react at 0 °C for 5 minutes. After detecting the complete reaction of the starting material by TLC, dilute the reaction solution with dichloromethane, extract with dichloromethane, saturated sodium bicarbonate solution and saturated brine, dry the organic layer over anhydrous sodium sulfate, concentrate under reduced pressure, and purify by semi-preparative chromatography to obtain the compound with a hydroxyl group at the 1-position.

[0075] General method for acyl group removal: Dissolve the compound (1.0 equiv) in a methanol / dichloromethane solution (reaction concentration 0.1 mol / L, 1 / 1, v / v), add sodium methoxide (0.2 equiv / each acyl group), react at room temperature for 2 hours, then add hydrogen ion exchange resin until pH = 7, filter and concentrate under reduced pressure, and purify by silica gel column chromatography to obtain the target compound.

[0076] General method for catalytic hydrogenation: Dissolve the compound (1.0 equiv) in a mixed solution of chromatographic grade methanol / dichloromethane / water (2 mL / 2 mL / 0.2 mL, 10 / 10 / 1, v / v / v), add an excess of palladium on carbon, displace with hydrogen for 20 minutes, and stir under hydrogen at room temperature for 2 days. Transfer the reaction solution with distilled water, filter the reaction mixture, concentrate, and purify by column chromatography on a Sep-Pak cartridge C18 column, collect the aqueous phase to obtain the fully deprotected oligosaccharide.

[0077] Experimental Example 1

[0078] Synthesis of arabinose building blocks 26 and 29, the synthetic route is as follows:

[0079]

[0080] Specific experimental operations and steps:

[0081] Compound 21: Dissolve compound 20 (1.058 g, 4.12 mmol) in dimethylformamide (20 mL), place the reaction system in a low-temperature reaction zone at -20 °C, add 2,6-dimethylpyridine (1.750 mL, 10.30 mmol) and di-tert-butylbis(trifluoromethanesulfonyloxy)silane (2.20 mL, 4.53 mmol), react at -20 °C for 2.5 hours. After detecting the complete reaction of the starting material by TLC, quench the reaction with methanol, extract the reaction solution with dichloromethane / saturated sodium bicarbonate solution, filter the solution, dry the organic layer over anhydrous sodium sulfate, concentrate under reduced pressure, and purify by silica gel column chromatography (PE / EA = 10 / 1) to obtain compound 21 as a white solid (1.153 g, 2.90 mmol, 71%). 1 H NMR(500MHz,CDCl 3)δ = 7.44 - 7.39 (m, 2H, Ar), 7.12 (d, J = 7.8 Hz, 2H, Ar), 5.24 (d, J = 5.9 Hz, 1H, H-1), 4.35 - 4.30 (m, 1H, H-5), 4.13 (dd, J = 7.5, 5.9 Hz, 1H, H-2), 4.00 (dd, J = 9.2, 7.4 Hz, 1H, H-3), 3.94 (dd, J = 10.3, 8.8 Hz, 1H, H-5), 3.90 - 3.85 (m, 1H, H-4), 2.32 (s, 3H, STol-CH 3 ), 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 carried out overnight. After the reaction of the starting material was complete as detected by TLC, the reaction solution was diluted with dichloromethane 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. It was separated and purified by silica gel column chromatography (PE / EA = 30 / 1) to obtain the target compound 22 (1.38 g, 15.24 mmol, 97%). 1 H NMR (500 MHz, CDCl 3 )δ = 8.09 (dd, J = 8.2, 1.4 Hz, 2H, Ar), 7.65 - 7.56 (m, 1H, Ar), 7.52 - 7.40 (m, 4H, Ar), 7.12 (d, J = 7.9 Hz, 2H, Ar), 5.51 (dd, J = 7.0, 4.7 Hz, 1H, H-2), 5.37 (d, J = 4.7 Hz, 1H, H-1), 4.39 (dd, J = 8.9, 4.8 Hz, 1H, H-5), 4.32 - 4.28 (m, 1H, H-4), 4.07 (td, J = 9.9, 4.8 Hz, 1H, H-5), 4.01 (d, J = 9.0 Hz, 1H, H-3), 2.31 (s, 3H, STol-CH 3 ), 1.01 (d, J = 28.8 Hz, 18H, t-Bu).

[0083] Compound 23: Compound 22 (7.873 g, 15.7 mmol) was subjected to the general method for removing silyl ether protecting groups to obtain compound 23 as a pale yellow syrup (5.65 g, 15.7 mmol, quant.) 1 H NMR (500 MHz, CDCl 3)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-CH 3 ).

[0084] Compound 24: Dissolve compound 23 (490 mg, 1.36 mmol) in dichloromethane (15 mL). Add imidazole (232 mg, 3.4 mmol) and tert-butyldimethylchlorosilane (308 mg, 2.04 mmol) at 0 °C and react at room temperature for 2 hours. After detecting the complete reaction of the raw materials by TLC, dilute the reaction solution with dichloromethane. Extract the reaction solution with dichloromethane, saturated sodium bicarbonate solution and saturated brine. Dry the organic layer with anhydrous sodium sulfate, concentrate under reduced pressure, and purify by silica gel column chromatography (PE / EA = 10 / 1) to obtain the target compound 24 (450 mg, 1.02 mmol, 75%). (450 mg, 1.02 mmol, 75%). 1 H NMR(400MHz, CDCl 3 )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-CH 3 ), 0.87(s, 9H, TBDMS-CH 3 ), 0.06(d, J=2.0Hz, 6H, TBDMS-CH 3 ). 13 C NMR(101MHz, CDCl 3)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), 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 reaction was carried out at room temperature for 30 h. After TLC detection showed that the raw materials had completely reacted, the reaction solution was diluted with dichloromethane, filtered through diatomaceous earth, and then 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 silica gel column chromatography (PE / EA = 30 / 1) to obtain Compound 25 as a transparent syrup (3.407 g, 6.03 mmol, 69%). 1 H NMR (500 MHz, CDCl 3 )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, 6H, Ar), 7.10 (d, J = 7.9 Hz, 2H, Ar), 5.60 (d, J = 1.6 Hz, 1H, H-1), 5.57 (d, J = 1.8 Hz, 1H, H-2), 4.80 (d, J = 12.0 Hz, 1H, Ar-CH 2 ), 4.64 (d, J = 12.0 Hz, 1H, Ar-CH 2 ), 4.44 (q, J = 4.7 Hz, 1H, H-4), 4.14 (dd, J = 5.5, 1.9 Hz, 1H, H-3), 3.80 (d, J = 4.5 Hz, 2H, H-5), 2.31 (s, 3H, STol-CH 3 ), 0.84 (s, 9H, TBS-CH 3 ), 0.00 (d, J = 4.4 Hz, 6H, TBS-CH 3 ). 13 C NMR (101 MHz, CDCl 3)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, 127.8, 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 the general method for the removal of silyl ether protecting groups to give Compound 26 as a transparent syrup (396 mg, 0.88 mmol, 98%). 1 HNMR (500 MHz, CDCl 3 )δ = 8.06 - 7.97 (m, 2H, Ar), 7.58 (t, J = 7.4 Hz, 1H, Ar), 7.44 (dt, J = 7.8, 3.7 Hz, 4H, Ar), 7.32 (ddd, J = 26.8, 18.5, 7.1 Hz, 5H, Ar), 7.12 (d, J = 7.8 Hz, 2H, Ar), 5.61 (s, 1H, H1), 5.56 (d, J = 1.7 Hz, 1H, H-2), 4.84 (d, J = 11.9 Hz, 1H, Ar-CH 2 ), 4.63 (d, J = 11.9 Hz, 1H, Ar-CH 2 ), 4.49 (dt, J = 6.2, 3.4 Hz, 1H, H-4), 4.16 (dd, J = 5.6, 1.8 Hz, 1H, H-3), 3.89 (dd, J = 12.3, 3.0 Hz, 1H, H-5), 3.70 (dd, J = 12.2, 3.9 Hz, 1H, H-5), 2.32 (s, 3H, STol-CH 3 ).

[0087] Compound 27: Compound 25 (840 mg, 1.49 mmol) was subjected to the general method for the hydrolysis of thioglycosides to give Compound 27 as a transparent syrup (594 mg, 1.3 mmol, 87%).

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

[0089] Compound 29: Dissolve compound 28α (54 mg, 0.086 mmol) in a mixed solution of methanol / dichloromethane (1.2 mL / 0.3 mL, 1 / 4, v / v). Add acetyl chloride (60 μL, 0.86 mmol). React at room temperature for 45 min. After detecting the complete reaction of the starting materials by TLC, dilute the reaction solution with dichloromethane, extract with dichloromethane, saturated sodium bicarbonate solution and saturated brine, dry the organic layer over anhydrous sodium sulfate, concentrate under reduced pressure and purify by silica gel column chromatography (PE / EA = 8 / 1) to obtain compound 29 as a transparent syrup (37.6 mg, 73.1 μmol, 85%). 1HNMR(500MHz,CDCl 3 ). δ = 8.07 - 8.03 (m, 2H, Ar), 7.96 (dd, J = 8.0, 1.4 Hz, 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, 1.3 Hz, 1H, Ar), 6.59 (s, 1H, H-1), 5.69 (d, J = 1.6 Hz, 1H, H-2), 4.90 (d, J = 11.7 Hz, 1H, Ar-CH 2 ), 4.67 (d, J = 11.8 Hz, 1H, Ar-CH 2 ), 4.48 (dt, J = 5.0, 3.5 Hz, 1H, H-4), 4.24 (dt, J = 5.3, 1.2 Hz, 1H, H-3), 3.92 (dd, J = 12.4, 3.0 Hz, 1H, H-5), 3.76 - 3.69 (m, 1H, H-5), 1.53 (tt, J = 7.8, 5.3 Hz, 1H, ABz-CH), 0.88 - 0.84 (m, 4H, ABz-CH 2 ).

[0090] Experimental Example 2

[0091] Synthesis of galacturonic acid sugar building block 32, the synthetic route is as follows:

[0092]

[0093] Compound 31: Dissolve compound 30 (3.296 g, 6.67 mmol) in dichloromethane (34 mL), and successively add dicyclohexylcarbodiimide (2.75 g, 13.3 mmol), levulinic acid (1.36 mL, 13.3 mmol), 4-dimethylaminopyridine (1.625 g, 13.3 mmol), and react at room temperature for 24 hours. After monitoring the reaction to completion by TLC, the reaction solution is extracted with dichloromethane, saturated sodium bicarbonate solution, and saturated brine solution. The organic layer is dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by medium-pressure preparative chromatography (PE / EA = 7 / 3) to obtain compound 31 (3.925 g, 6.62 mmol, 99%).

[0094] Compound 32: Compound 31 (711.5 mg, 1.20 mmol) was subjected to the general method of hydrolysis with 4-methoxyphenyl to obtain the hemiacetal intermediate compound (446.1 mg, 0.92 mmol, 76%). The hemiacetal intermediate compound (904 mg, 1.525 mmol) was dissolved in anhydrous dichloromethane (10 mL) after vacuum drying. At 0 °C, 2,2,2-trifluoro-N-phenyliminoacetyl 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. The reaction was carried out at room temperature for 2 hours. After TLC detection showed that the reaction was complete, the reaction system was concentrated under reduced pressure at 30 °C, and then separated and purified by medium-pressure semi-preparative chromatography column (PE / EA = 5 / 1) to obtain Compound 32 (1.044 g, 1.5 mmol, quant.).

[0095] Experimental Example 3

[0096] Synthesis of rhamnose building blocks 35 and 38. The synthetic route is as follows:

[0097]

[0098] Compound 34: Compound 33 (6.63 g, 21.36 mmol) was dissolved in dimethylformamide (107 mL). At 0 °C, sodium hydride (1.71 g, 42.8 mmol) was added, and then 2-bromomethylnaphthalene (9.45 g, 42.8 mmol) was added. The reaction was stirred at room temperature. After TLC detection showed that the reaction was over, the reaction was terminated by adding distilled water in an ice-water bath. The mixture was extracted with distilled water and ethyl acetate, and 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% acetic acid aqueous solution (25 mL), and heated under reflux at 60 °C 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), and dibutyltin oxide (11.65 g, 46.8 mmol) was added. The reaction was heated under reflux at 120 °C for 4 hours. After returning to room temperature, half of the solvent was removed by rotary evaporation. Acetonitrile (100 mL) was added, and then cesium fluoride (7.109 g, 46.8 mmol) and benzyl bromide (8.0 g, 46.8 mmol) were added in sequence. The reaction was heated under reflux at 80 °C for 12 h. After TLC detection showed that the reaction was over, it was transferred with dichloromethane, and insoluble impurities were removed by filtration through diatomaceous earth. The filtrate 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%). 1 H NMR(500MHz,CDCl 3) δ = 7.87 - 7.72 (m, 4H, Ar), 7.52 - 7.41 (m, 4H, Ar), 7.40 - 7.28 (m, 8H, Ar), 7.10 (d, J = 7.8 Hz, 2H, Ar), 5.46 (s, 1H, H-1), 5.04 (d, J = 11.2 Hz, 1H, Ar-CH 2 ), 4.81 (d, J = 11.1 Hz, 1H, Ar-CH 2 ), 4.72 (s, 2H, Ar-CH 2 ), 4.28 - 4.20 (m, 2H, H-2, H-5), 3.90 (dd, J = 9.1, 3.2 Hz, 1H, H-3), 3.58 (t, J = 9.3 Hz, 1H, H-4), 2.31 (s, 3H, STol-CH 3 ), 1.32 (s, 3H, H-6). 13 C NMR (101 MHz, CDCl 3 ) 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 protection, dissolve compound 34 (4.3 g, 8.8 mmol) in pyridine (95 ml), add acetic anhydride (1.65 mL, 17.6 mmol) and 4-dimethylaminopyridine (215 mg, 1.76 mmol) at 0 °C, stir at 0 °C for 0.5 hour, and then react at room temperature for 2 hours. When the reaction is completed as detected by TLC, transfer the reaction system with ethyl acetate, extract it with 1 M hydrochloric acid and saturated sodium bicarbonate respectively, dry the organic phase with anhydrous sodium sulfate to remove water, concentrate and spin-dry it, and purify it by silica gel column chromatography (PE / EA = 15 / 1) to obtain compound 35 (4.64 g, 8.7 mmol, 99%). 1 HNMR (500 MHz, CDCl 3 ) δ = 7.77 - 7.68 (m, 3H), 7.63 (s, 1H, Ar), 7.39 (tt, J = 6.6, 3.7 Hz, 2H, Ar), 7.33 (dd, J = 7.9, 5.8 Hz, 3H, Ar), 7.29 - 7.25 (m, 2H, Ar), 7.25 - 7.16 (m, 4H, Ar), 7.04 (d, J = 7.8 Hz, 2H, Ar), 5.73 (d, J = 3.3 Hz, 1H, H-2), 4.97 (d, J = 11.0 Hz, 1H, Ar-CH 2), 4.76 - 4.67 (m, 3H, H1, Ar-CH 2 ), 4.42 (d, J = 11.1 Hz, 1H, Ar-CH 2 ), 3.57 (dd, J = 9.2, 3.4 Hz, 1H, H-3), 3.43 (t, J = 9.2 Hz, 1H, H-4), 3.38 - 3.29 (m, 1H, H-5), 2.25 (s, 3H, STol-CH 3 ), 2.16 (s, 3H, Ac-CH 3 ), 1.34 (d, J = 6.1 Hz, 3H, H-6).

[0100] Compound 37: Under nitrogen protection, 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, and then 2-bromomethylnaphthalene (1.08 g, 4.88 mmol) was added. The ice-water bath was removed, and the reaction was stirred at room temperature. The reaction was monitored by TLC. After the reaction was completed, the reaction was terminated by adding distilled water under an ice-water bath. The mixture was extracted with distilled water and ethyl acetate, and the organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure to obtain an intermediate compound. The intermediate compound was then dissolved in 80% acetic acid aqueous solution (25 mL), heated under reflux at 60 °C for 5 hours. The reaction system was extracted with dichloromethane and water. The organic phase was filtered and dried over anhydrous sodium sulfate, concentrated to dryness by rotary evaporation, and purified by silica gel column chromatography (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 reaction was heated under reflux at 120 °C for 5 h. After returning to room temperature, half of the solvent was removed by rotary evaporation. Acetonitrile (6 mL) was added, and then benzyl bromide (539 μL, 4.54 mmol) and tetrabutylammonium bromide (1.46 g, 4.54 mmol) were added in sequence. The reaction was heated under reflux at 80 °C for 12 h. The reaction was monitored by TLC. After the reaction was completed, the reaction mixture was transferred with dichloromethane, filtered through diatomaceous earth to remove insoluble impurities, concentrated under reduced pressure, and purified by silica gel column chromatography (PE / EA = 7 / 1) to obtain compound 38 (768 mg, 1.77 mmol, 78%).

[0102] Experimental Example 4

[0103] The synthesis of Lycium ruthenicum Murr. oligosaccharides 41, 44, 47, 51 and 53 is as follows:

[0104]

[0105] Compound 40: Glycosyl donor 39 (241 mg, 0.4 mmol) and acceptor 26 (130 mg, 0.29 mmol) were used in the general glycosylation method A to obtain compound 40 (144 mg, 0.17 mmol, 59%). 1 HNMR (500 MHz, CDCl 3 ) δ = 8.03 (dd, J = 8.3, 1.3 Hz, 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 = 7.9 Hz, 2H, Ar), 5.66 - 5.60 (m, 1H, A - H1), 5.57 (t, J = 1.8 Hz, 1H, A - H2), 5.38 (d, J = 1.5 Hz, 1H, D - H2), 5.27 (s, 1H, D - H1), 4.79 (d, J = 11.9 Hz, 1H, Ar - CH 2 ), 4.65 - 4.61 (m, 2H, Ar - CH 2 ), 4.59 (q, J = 4.3 Hz, 1H, D - H3), 4.53 (d, J = 12.1 Hz, 1H, Ar - CH 2 ), 4.46 (dd, J = 12.2, 4.6 Hz, 2H, Ar - CH 2 ), 4.27 (ddd, J = 5.4, 2.0, 0.9 Hz, 1H, A - H3), 4.20 (td, J = 5.2, 3.7 Hz, 1H, A - H4), 4.01 - 3.90 (m, 2H, D - H4, D - H5), 3.78 (dd, J = 11.4, 4.1 Hz, 1H, D - H5), 3.59 (dd, J = 10.7, 3.7 Hz, 1H, A - H5), 3.52 (dd, J = 10.8, 5.0 Hz, 1H, A - H5), 2.29 (s, 3H, STol - CH 3 ). 13 CNMR (126 MHz, CDCl 3 ) 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 successively subjected to the general method for thioglycoside hydrolysis, the general method for acyl group removal, and the general method for catalytic hydrogenation to give Compound 41 (7 mg, 0.042 mmol, 29%). 1 HNMR(600MHz,D 2 O)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,D 2 O)δ=107.4,96.8,92.6,83.9,80.9,76.6,72.4,72.2,71.9,70.9,70.5,70.2,69.3,68.8,68.6,68.1,66.4,63.0,62.5,61.2.HR-ESI-MS(m / z):calcd for C 10 H 18 O 9 Na + (M+Na) + :305.0843,found:305.0822.

[0107] Compound 43: The glycosyl donor 39 (140 mg, 0.23 mmol) and the acceptor 42 (59 mg, 0.116 mmol) were reacted according to General Glycosylation Method A to give Compound 87 (85 mg, 0.09 mmol, 78%). 1 HNMR(500MHz,CDCl 3)δ = 8.01 - 7.95 (m, 2H, Ar), 7.60 - 7.53 (m, 2H, Ar), 7.49 (tt, J = 7.5, 1.4 Hz, 1H, Ar), 7.23 - 7.15 (m, 9H, Ar), 5.76 (t, J = 9.8 Hz, 1H, A - H2), 5.37 (s, 1H, D - H2), 5.29 (d, J = 1.3 Hz, 1H, D - H1), 5.08 (d, J = 11.7 Hz, 1H, Ar - CH2), 4.67 (d, J = 11.9 Hz, 1H, Ar - CH2), 4.59 - 4.40 (m, 8H, Ar - CH2, A - H1, D - H3), 4.12 (dd, J = 9.7, 2.8 Hz, 1H, A - H3), 4.00 (ddd, J = 7.8, 3.3, 1.0 Hz, 1H, A - H4), 3.94 (dt, J = 5.0, 1.0 Hz, 1H, D - H5), 3.76 (ddd, J = 6.8, 5.5, 1.1 Hz, 1H, D - H5), 3.67 - 3.54 (m, 4H, D - H4, A - H5, A - H6), 2.80 - 2.64 (m, 2H, SEt - CH 2 ), 1.22 (dt, J = 18.6, 7.5 Hz, 3H, SEt - CH 3 ). 13 C NMR (126 MHz, CDCl 3 )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 successively subjected to the general method for thioglycoside hydrolysis, the general method for acyl group removal, and the general method for catalytic hydrogenation to give Compound 44 (12 mg, 0.0178 mmol, 48%). 1 H NMR (600 MHz, D 2O) δ 5.12 (d, 0.4H), 5.08 (dd, J = 4.6, 1.6 Hz, 1H), 4.48 (d, J = 7.9 Hz, 1H), 4.05 (dt, J = 4.0, 2.0 Hz, 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). 13 C NMR (151 MHz, D 2 O) δ 109.2, 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): calcd for C 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 reacted according to the general glycosylation method B to give compound 46 (161 mg, 0.156 mmol, 50%). 1 H NMR (500 MHz, CDCl 3) δ 8.18 - 8.12 (m, 2H, Ar), 7.47 (t, J = 7.4 Hz, 1H, Ar), 7.40 - 7.25 (m, 17H, Ar), 7.21 (ddd, J = 17.3, 7.4, 3.0 Hz, 11H, Ar), 7.11 - 7.07 (m, 2H, Ar), 6.86 - 6.79 (m, 2H, Ar), 5.68 (dd, J = 10.2, 3.9 Hz, 1H, D - H2), 5.26 (dd, J = 13.6, 3.9 Hz, 1H, D - H1), 4.98 (dd, J = 11.1, 7.4 Hz, 2H, Ar - CH2), 4.85 (dd, J = 11.8, 5.3 Hz, 2H, Ar - CH2), 4.76 (d, J = 7.5 Hz, 1H, A - H1), 4.69 (d, J = 12.0 Hz, 1H, Ar - CH2), 4.63 - 4.51 (m, 5H, Ar - CH2, D - H4, A - H4), 4.24 (d, J = 1.6 Hz, 2H, Ar - CH2), 4.17 - 4.10 (m, 2H, D - H3, A - H5), 3.95 - 3.83 (m, 2H, A - H2, D - H), 3.77 (s, 3H, COOMe - CH 3 ), 3.62 (t, J = 8.5 Hz, 1H, D - H), 3.49 (dd, J = 9.8, 2.9 Hz, 1H, A - H3), 3.41 (dd, J = 8.6, 5.2 Hz, 1H, D - H), 3.25 (s, 3H, MP - CH 3 ). 13 C NMR (126 MHz, CDCl 3 ) δ 167.8, 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 of 4-methoxyphenyl hydrolysis 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 of acyl group 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 of catalytic hydrogenation to obtain Compound 47 (8 mg, 0.022 mmol, 90%). 1 HNMR(600MHz,D 2 O)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,D 2 O)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 subjected to the general glycosylation method A to obtain Compound 48 (843 mg, 0.93 mmol, 73%). 1 HNMR(400MHz,CDCl 3 )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 - CH 2 ),5.18(dd,J=10.3,1.7Hz,1H,All - CH 2), 5.05 - 5.01 (m, 2H, D - H1, Ar - CH 2 ), 4.84 (d, J = 2.0 Hz, 1H, D - H2), 4.80 (dd, J = 5.8, 2.3 Hz, 2H, A - H1, Ar - CH 2 ), 4.72 (s, 1H, Ar - CH 2 ), 4.69 (s, 1H, Ar - CH 2 ), 4.63 (s, 1H, Ar - CH 2 ), 4.60 (s, 1H, Ar - CH 2 ), 4.50 (d, J = 11.1 Hz, 1H, Ar - CH 2 ), 4.18 - 4.12 (m, 1H, All - CH 2 ), 4.09 (t, J = 2.5 Hz, 1H, All - CH 2 ), 4.02 (dd, J = 10.1, 3.4 Hz, 1H, A - H2), 3.97 - 3.90 (m, 2H, Ar - CH 2 , A - H3), 3.85 - 3.73 (m, 3H, D - H5, D - H3, A - H5), 3.54 (s, 3H, Me), 3.50 (d, J = 9.5 Hz, 1H, A - H4), 2.71 - 2.55 (m, 4H, Lev - CH 2 ), 2.13 (s, 2H, Lev - CH 3 ), 1.38 (d, J = 6.2 Hz, 3H, A - H6). 13 C NMR (101 MHz, CDCl 3 ) δ 206.0, 171.5, 168.2, 138.6, 138.2, 137.9, 135.9, 133.8, 133.3, 133.0, 128.3, 128.3, 128.2, 128.2, 128.1, 128.1, 128.0, 127.9, 127.8, 127.7, 127.58, 127.57, 127.5, 127.4, 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: Dissolve compound 48 (228 mg, 0.25 mmol) in pyridine / acetic acid (3.2 mL / 0.8 mL, 4 / 1). Add 80% hydrazine hydrate under an ice bath and stir for 3 h. After the reaction is completed as detected by TLC, transfer the system with EtOAc, and then extract with 1 M HCl and saturated sodium bicarbonate solution respectively. Combine the organic phases, dry over anhydrous sodium sulfate, concentrate, and purify by silica gel column chromatography (EA / EA system, EA% = 30%) to obtain compound 49 (191 mg, 96%). 1 HNMR(500MHz,CDCl 3 )δ7.84 - 7.75(m,4H,Ar),7.45 - 7.25(m,22H,Ar),5.95 - 5.81(m,1H,All-CH),5.25(ddd,J=17.2,10.5,2.0Hz,1H,All-CH 2 ),5.20 - 5.14(m,1H,All-CH 2 ),5.05(dd,J=11.1,2.9Hz,1H,Ar-CH 2 ),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-CH 2 ),4.77(d,J=9.7Hz,1H,D-H2),4.73 - 4.69(m,4H,A-H2,Ar-CH 2 ),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-CH 2 ),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: Dissolve compound 49 (45 mg, 0.056 mmol) in 2 mL of CH 3 COOH, add Pd(PPh 3 ) 4 (19 mg, 0.017 mmol), react at 80 °C for 1 h. After detecting the end of the reaction by TLC, transfer the system with DCM, add solid NaHCO 3 to neutralize acetic acid, then extract successively with distilled water and saturated NaCl solution. The organic phase is dried with anhydrous sodium sulfate, concentrated and the solvent is rotary evaporated, and purified by silica gel column chromatography (DCM / MeOH = 10 / 1) to obtain compound 50 (33 mg, 0.043 mmol, 77%).

[0114] Compound 51: Using the general method of catalytic hydrogenation for compound 50 (34 mg, 0.044 mmol), compound 51 (12 mg, 0.034 mmol, 77%) is obtained. 1 HNMR(600MHz,D 2 O)δ5.06(dd,J=4.3,2.7Hz,1H),4.95(d,J=3.9Hz,0.7H,A-H-1β),4.91(dd,J=5.9,1.6Hz,1H),4.79(s,0.3H,A-H-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,D 2 O)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: Dissolve compound 50 (50 mg, 0.065 mmol) in a mixed solvent of MeOH / H 2 O (1 mL / 1 mL, 1 / 1, v / v). Add LiOH-hydrate (0.1 M / L, 27 mg, 0.65 mmol), and react at room temperature for 5 h. After the reaction is completed as detected by TLC, adjust the pH to slightly acidic with hydrogen ion exchange resin, filter, concentrate and evaporate the solvent to dryness, and purify 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) is obtained by the general method of catalytic hydrogenation to give compound 53 (12 mg, 0.035 mmol, quant.). 1 HNMR (600 MHz, D 2 O) δ 5.08 (s, 2H), 4.79 (s, 1H), 4.42 (dd, J = 21.3, 7.6 Hz, 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, 7.1 Hz, 11H), 3.61 - 3.45 (m, 3H), 3.37 - 3.23 (m, 5H), 3.09 - 3.00 (m, 2H), 1.13 (d, J = 6.3 Hz, 9H). 13 C NMR (151 MHz, D 2 O) δ 101.2, 93.9, 92.8, 91.5, 81.5, 80.9, 79.9, 76.5, 75.0, 72.6, 72.5, 71.2, 70.7, 70.4, 68.9, 68.5,, 68.3, 66.9, 16.7. HR-ESI-MS (m / z): calcd for C 12 H 20 O 11 Na + (M + Na) + : 363.0898, found: 363.2270.

[0116] Experimental Example 5

[0117] Synthesis of Lycium ruthenicum oligoglycosides 55, 57, 59 and 61. The synthetic route is as follows:

[0118]

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

[0120] (mmol) According to the general glycosylation method A, compound 54 (62 mg, 0.067 mmol, 40%) was obtained. 1 H NMR (500 MHz, CDCl 3 ) δ 8.11 - 8.03 (m, 2H, Ar), 8.00 - 7.92 (m, 3H, Ar), 7.57 (td, J = 7.7, 3.0 Hz, 2H, 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 - H1), 5.69 (d, J = 1.4 Hz, 1H, A - H2), 5.36 (d, J = 1.4 Hz, 1H, D - H2), 5.28 (s, 1H, D - H1), 4.86 (d, J = 11.7 Hz, 1H, Ar - CH 2 ), 4.68 (d, J = 11.8 Hz, 1H, Ar - CH 2 ), 4.64 - 4.58 (m, 2H, D - H3, Ar - CH 2 ), 4.52 (d, J = 12.1 Hz, 1H, Ar - CH 2 ), 4.45 (dd, J = 15.4, 12.1 Hz, 2H, Ar - CH 2 ), 4.33 (d, J = 4.8 Hz, 1H, A - H3), 4.23 (q, J = 4.8 Hz, 1H, D - H4), 4.00 (dd, J = 11.3, 4.2 Hz, 1H, D - H5), 3.94 (d, J = 5.4 Hz, 1H, A - H4), 3.77 (dd, J = 11.3, 4.9 Hz, 1H, D - H5), 3.59 (dd, J = 10.8, 3.7 Hz, 1H, A - H5), 3.53 (dd, J = 10.8, 5.0 Hz, 1H, A - H5), 1.51 (tt, J = 8.0, 5.2 Hz, 1H, ABz - CH), 0.85 (tt, J = 8.2, 2.8 Hz, 4H, ABz - CH 2 ).

[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, added MS, and then the catalyst PPh 3 AuOTf (9 mg, 0.015 mmol) was added and stirred at room temperature for 16 h. After the reaction was monitored by TLC and completed, triethylamine was added to quench the reaction, and DCM / saturated NaHCO 3The solution was extracted, and the organic phase was dried with anhydrous sodium sulfate, concentrated by rotary evaporation, and purified by column chromatography (PE / EA = 5 / 1) to obtain compound 55 (9 mg, 0.075 mmol, 30%). 1 H NMR (500 MHz, CDCl 3 ) δ 8.02 (t, J = 7.6 Hz, 4H), 7.95 (d, J = 7.7 Hz, 2H), 7.55 (dt, J = 14.1, 7.5 Hz, 3H), 7.45 - 7.32 (m, 10H), 7.22 (ddt, J = 28.5, 20.9, 6.3 Hz, 20H), 7.07 (d, J = 7.9 Hz, 2H), 5.62 (s, 1H), 5.57 (s, 1H), 5.38 (d, J = 24.3 Hz, 2H), 5.25 (s, 2H), 4.78 (d, J = 11.9 Hz, 1H), 4.72 - 4.35 (m, 7H), 4.27 (d, J = 5.6 Hz, 1H), 4.20 (q, J = 4.4 Hz, 1H), 4.15 (q, J = 4.8 Hz, 1H), 4.11 (d, J = 5.6 Hz, 1H), 3.96 (dd, J = 11.5, 4.1 Hz, 1H), 3.91 (d, J = 5.5 Hz, 1H), 3.86 (dd, J = 11.3, 4.0 Hz, 1H), 3.76 (dd, J = 11.4, 4.1 Hz, 1H), 3.68 (dd, J = 11.4, 3.8 Hz, 1H), 3.56 (dd, J = 10.8, 3.8 Hz, 1H), 3.49 (dd, J = 10.7, 5.0 Hz, 1H), 2.28 (s, 2H).

[0122] Compound 57: Compound 45 (27 mg, 0.042 mmol) was dissolved in DCM (1 mL), 4A molecular sieve was added, and then BSP (10.55 mg, 0.048 mmol) and TTBP (15.65 mg, 0.063 mmol) were added. It was placed in a low-temperature reaction zone at -60 °C, and Tf 2 O (9 μL, 0.052 mmol) was added and the reaction was carried out at -60 °C for 30 min. After TLC detection showed that the raw materials had completely reacted, 1-octene (8.5 μL, 0.052 mmol) was added, and the reaction was carried out at -60 °C for 15 min. Then the temperature was lowered to -78 °C, and compound 56 (27 mg, 0.053 mmol) dissolved in 1 mL of DCM was added, and the reaction was carried out at -78 °C for 3 h. After TLC detection showed that the raw materials had completely reacted, triethyl phosphite (22 μL, 0.126 mmol) was added, and the reaction was carried out at -78 °C for 1 h, and then the temperature was restored to room temperature. The reaction solution was extracted with DCM / saturated NaHCO3 solution and saturated NaCl solution, and the organic layer was washed with Na 2 SO 4Dried, concentrated under reduced pressure, and purified by silica gel column chromatography (PE / EA = 5 / 1, v / v) to obtain Compound 13 (17 mg, 0.015 mmol, 37%). 1 HNMR(400MHz,CDCl 3 )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 - 3.57(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 subjected to the general glycosylation method B to obtain Compound 59 (85 mg, 0.088 mmol, 49%). 1 HNMR(500MHz,CDCl 3 )δ7.86 - 7.52(m,3H,Ar),7.45 - 7.05(m,34H,Ar),5.31(t,J=2.5Hz,1H,D - H2),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 - CH 3 ),1.19(d,J=5.9Hz,3H,A - H6).ESI - MS(m / z):calcd for C60 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 used in the general glycosylation method A to obtain compound 60 (85 mg, 0.09 mmol, 67%, α / β = 2.3 / 1). 1 H NMR δ1α (400 MHz, CDCl 3 ) δ7.87 - 7.77 (m, 4H, Ar), 7.48 (ddd, J = 9.6, 6.9, 2.1 Hz, 3H, Ar), 7.36 - 7.28 (m, 13H, Ar), 7.22 - 7.09 (m, 5H, Ar), 6.90 (d, J = 8.1 Hz, 1H, Ar), 5.83 (dd, J = 3.5, 1.7 Hz, 1H, D - H4), 5.35 (d, J = 1.7 Hz, 1H, A - H2), 5.07 (d, J = 11.2 Hz, 1H, Ar - CH 2 ), 4.98 (dd, J = 17.3, 2.6 Hz, 2H, D - H5, Ar - CH 2 ), 4.83 (dd, J = 16.0, 11.5 Hz, 2H, Ar - CH 2 ), 4.71 (d, J = 11.8 Hz, 2H, Ar - CH 2 ), 4.64 (d, J = 12.0 Hz, 1H, Ar - CH 2 ), 4.52 (dd, J = 15.7, 11.5 Hz, 2H, Ar - CH2), 4.32 - 4.29 (m, 1H, A - H1), 4.21 (dd, J = 9.4, 6.1 Hz, 1H, A - H5), 4.00 (dd, J = 10.0, 3.4 Hz, 1H, D - H3), 3.88 (dd, J = 9.5, 3.0 Hz, 1H, A - H3), 3.79 (dd, J = 10.1, 3.5 Hz, 1H, D - H2), 3.59 (s, 3H, Me), 3.51 (s, 1H, A - H4), 2.70 - 2.53 (m, 4H, Lev - CH 2 ), 2.36 (s, 3H, Lev - CH 3 ), 2.14 (s, 3H, STol - CH 3 ), 1.41 (d, J = 6.2 Hz, 3H, A - H6). 13 C NMR (101 MHz, CDCl3 ) δ 206.0, 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 reacted according to the general glycosylation method B to obtain compound 16 (8 mg, 0.088 mmol, 24%). 1 1H NMR (500 MHz, CDCl 3 ) δ 7.74 (dt, J = 11.1, 5.7 Hz, 3H), 7.46 - 7.06 (m, 39H), 5.73 (dt, J = 15.4, 3.0 Hz, 1H), 5.57 (d, J = 3.7 Hz, 1H), 5.05 (d, J = 1.7 Hz, 1H), 4.91 (dd, J = 19.5, 11.6 Hz, 2H), 4.81 (d, J = 11.5 Hz, 1H), 4.76 - 4.36 (m, 13H), 4.23 (d, J = 12.8 Hz, 1H), 4.04 (d, J = 12.7 Hz, 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 = 6.1 Hz, 3H).

[0126] Application Example 1 Anti - pancreatic cancer activity test of Lycium ruthenicum oligose and its derivatives

[0127] Specific test operations and steps:

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

[0129] Take pancreatic cancer cells PANC - 1, BxPC - 3, and AsPC - 1 in good growth state (cell density is ~1×10 4The suspension was inoculated into a 96-well plate. 100 μL was added to each well, with 3 replicates set, and a blank group (equal volume of culture medium) and a control group (cells incubated with an equal volume of culture medium) were also set, and cultured overnight. Aqueous solutions of different concentrations of black wolfberry oligosaccharides and their derivatives 41, 44, 47, 51, and 53 were respectively added to the 96-well plate, and together with the blank group and the control group, cultured at 37 °C and 5% CO 2 under the conditions for 72 hours. 10 μL of thiazolyl blue (MTT) solution (5 mg / mL) was added, and the culture was continued in the incubator for 4 hours. The culture medium was discarded, 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 measured with an enzyme-linked immunosorbent assay (ELISA) reader. The cell survival rate was calculated according to the following formula: Cell survival rate = (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] Using 5 synthesized oligosaccharide fragments and the positive control drug gemcitabine, pancreatic cancer cells were treated at different concentrations, and the results were as Figure 1 、 2 、shown in Figure 3. Compounds 44 and 53 showed certain inhibitory effects on pancreatic cancer cells PANC-1, AsPC-1, and BxPC-3. Among them, the inhibition rates of compounds 44 and 53 on the gemcitabine-resistant cell PANC-1 reached 78.3% and 57.84% respectively, and the highest inhibition rates on cells AsPC-1 and BxPC-3 also reached 45.4% and 47.5% respectively. The most active compound 44 was selected for IC 50 testing, and the results were as Figure 4 shown in Figure. The IC 50 value of compound 44 for pancreatic cancer cell PANC-1 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 domains of black wolfberry polysaccharides, and larger oligosaccharides containing this structural fragment can be continuously synthesized in the future.

[0131] In summary, as can be seen from the examples, black wolfberry oligosaccharides can become potential carbohydrate drugs for the treatment of pancreatic cancer.

Claims

1. A black wolfberry oligosaccharide derivative, characterized in that: The derivative is a compound having a structure represented by formula (1) to (8) or a pharmaceutically acceptable salt thereof: 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, benzylidene acetal, and isopropylidene ketal; 13 Any one of hydrogen, methyl, ethyl, tert-butyl, benzyl, allyl, trichloroethyl, trimethylsilylethyl; PG 18 It is any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, levulinyl and allyloxycarbonyl.

2. The black wolfberry 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 black wolfberry oligosaccharide derivative according to claim 1, characterized in that: The steps include: S1. The glycosyl donor 9 and the glycosyl acceptor 10 are subjected to glycosylation reaction to obtain disaccharide 1; the acyl protecting group of disaccharide 1 is removed under alkaline conditions, and the aromatic protecting group is removed by reduction reaction to complete the deprotection to obtain a fully deprotected disaccharide 1a; the synthetic route is as follows: 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; S2. Disaccharide 1 selectively removes the protecting group PG4 to obtain disaccharide 11, which is then subjected to glycosylation reaction with glycosyl donor 9 under the catalytic action of a promoter to obtain trisaccharide 2; trisaccharide 2 removes the acyl protecting group under alkaline conditions, and removes the aromatic protecting group by reduction reaction to complete the deprotection and obtain fully deprotected trisaccharide 2a; the synthetic route is as follows: S3. The glycosyl donor 9 and the glycosyl acceptor 12 are subjected to glycosylation reaction to obtain disaccharide 3; the acyl protecting group of disaccharide 3 is removed under alkaline conditions, and the aromatic protecting group is removed by reduction reaction to complete the deprotection to obtain a fully deprotected trisaccharide 3a; the synthetic route is as follows: S4. The glycosyl donor 13 and the glycosyl acceptor 14 are subjected to glycosylation reaction to obtain disaccharide 4; the acyl protecting group of disaccharide 4 is removed under alkaline conditions, and the aromatic protecting group is removed by reduction reaction to complete the deprotection to obtain a fully deprotected trisaccharide 4a; the synthetic route is as follows: S5. The glycosyl donor 13 and the glycosyl acceptor 15 are subjected to glycosylation reaction to obtain disaccharide 5; the acyl protecting group of disaccharide 5 is removed under alkaline conditions, and the aromatic protecting group is removed by reduction reaction to complete the deprotection to obtain a fully deprotected trisaccharide 5a; the synthetic route is as follows: S6. The glycosyl donor 16 and the glycosyl acceptor 14 are subjected to glycosylation reaction to obtain disaccharide 6; the acyl protecting group of disaccharide 6 is removed under alkaline conditions, and the aromatic protecting group is removed by reduction reaction to complete deprotection to obtain fully deprotected trisaccharide 6a; the synthetic route is as follows: S7. The glycosyl donor 17 and the glycosyl acceptor 18 are subjected to glycosylation reaction, and the remote participation effect of the C4 acyl group is used to complete the construction of the 1,2-cis-glycosidic bond to obtain disaccharide 7; the acyl protecting group of disaccharide 7 is removed under alkaline conditions, and the aromatic protecting group is removed by reduction reaction to complete the deprotection and obtain the fully deprotected disaccharide 7a; the synthetic route is as follows: S8. The glycosyl donor 19 and the glycosyl acceptor 14 are subjected to glycosylation reaction to obtain trisaccharide 8; the acyl protecting group of trisaccharide 7 is removed under alkaline conditions, and the aromatic protecting group is removed by reduction reaction to complete the deprotection to obtain fully deprotected trisaccharide 8a; the synthetic route is as follows:

4. The method for preparing the black wolfberry oligosaccharide derivative according to claim 3, characterized in that: 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.

5. The method for preparing the black wolfberry oligosaccharide derivative according to claim 3, characterized in that: 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 black wolfberry oligosaccharide derivative according to claim 3, characterized in that: The molar ratio of the glycosyl donor and the glycosyl acceptor in the synthetic route is (1-2):1; the glycosylation reaction conditions are: dissolving the glycosyl donor and the glycosyl acceptor in a dry solvent, adding a molecular sieve and a promoter, stirring and reacting for 3-7 hours at a set temperature to obtain a 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; the stirring at the set temperature is stirring and reacting at room temperature of 25°C, or stirring and reacting at a mixture of ice and water at 0°C, or stirring and reacting at a mixture of ice and sodium chloride at -5 to -20°C, or stirring and reacting at a mixture of acetonitrile and dry ice at -40°C, or stirring and reacting at a mixture of acetone and dry ice at -60°C, or stirring and reacting at a mixture of acetone and dry ice at -78°C.

7. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the black wolfberry 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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