Three candidate antigen chimeric oligosaccharides of helicobacter pylori and preparation method of three candidate antigen chimeric oligosaccharides

Abstract

The invention discloses three candidate antigen chimeric oligosaccharides of helicobacter pylori and a preparation method of the three candidate antigen chimeric oligosaccharides. A mannoheptalactone compound is subjected to triethyl orthoacetate protection of hydroxyl groups at the 2 and 3 positions, reduction of heterotopic positions, addition of alkyne olefine acid ester at the heterotopic positions, ring opening of triethyl orthoacetate and protection of hydroxyl groups at the 3 positions by acetyl to obtain a mannoheptase alkyne olefine acid ester donor, and the mannoheptalactone compound is subjected to three candidate antigen chimeric oligosaccharides of helicobacter pylori and the three candidate antigen chimeric oligosaccharides of helicobacter pylori. Then carrying out glycosylation reaction with 1-azidopropanol, and further reducing an azide group into an amino group; glucose thioglycoside is subjected to 6-site hydroxyl protection through a photosensitive protecting group, iodinated acrylic acid is added to 1-site, glucose thioglycoside is coupled with 1-hexyne to obtain a glucose alkyne olefine acid ester donor protected by the photosensitive protecting group, alpha-1, 6 connected glucooligosaccharide is prepared through a photosensitive one-pot method, then 6-site hydroxyl is protected through azidopropyl, and an azido group is reduced into amino; glutaric acid is used as a connecting arm to connect the two fragments, and the target chimeric oligosaccharide is finally obtained through three steps of propylidene removal, acetyl removal and hydrogenation deprotection. The preparation method is mild in condition and simple to operate.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to three candidate antigen chimeric oligosaccharides of Helicobacter pylori and their preparation methods. Background Technology

[0002] Helicobacter pylori is a Gram-negative pathogen whose unique urease activity and flagellated motility enable it to colonize the gastric mucosa, inducing chronic inflammation and leading to peptic ulcers, gastric mucosa-associated lymphoid tissue lymphoma, and gastric cancer. Although eradication therapy using proton pump inhibitors (PPIs) combined with antibiotics is widely used, resistance rates to first-line drugs such as clarithromycin and levofloxacin have exceeded 30%, and in some areas even surpass 50%, resulting in a year-on-year increase in treatment failure rates. Therefore, developing new prevention and control measures that can overcome drug resistance limitations, especially vaccines with both preventative and therapeutic potential, has become a key breakthrough in solving this public health challenge.

[0003] Currently, the development of candidate antigens for Helicobacter pylori vaccines mainly focuses on bacterial virulence factors, urease, and CagA protein. For example, Malfertheiner's team conducted a trial combining three virulence factor components—CagA, VacA, and neutrophil activating protein (NAP)—as vaccine antigens. The results confirmed that the vaccine could induce an immune response, generating antigen-specific antibodies and forming durable T-cell immune memory. Based on this positive finding, researchers selected healthy volunteers for follow-up trials to verify the vaccine's preventive effect against CagA-positive Helicobacter pylori and its tolerability in humans. Unfortunately, the trial data showed no significant difference in preventive effect between the vaccine group and the placebo group, failing to achieve the expected goals. Zou Quanming et al., using recombinant DNA technology, successfully developed an oral recombinant Helicobacter pylori vaccine. This vaccine uses a fusion protein of the urease B subunit and the heat-labile enterotoxin B subunit as its core antigen and conducted a randomized, double-blind, placebo-controlled phase III clinical trial in children. The test results further confirmed that urease has the potential to be a candidate antigen for vaccines, but the oral formulation has the problem of excessively high antigen dosage, and its protective efficacy still needs to be tracked and verified over a longer period of time.

[0004] To further improve the efficacy and safety of vaccines, synthetic vaccines are gradually becoming a new research and development direction. In previous studies, we synthesized and screened a series of mannogen oligosaccharide antigens from lipopolysaccharide (LPS). We found that a Helicobacter pylori glycoconjugate vaccine made from mannogen trisaccharide could induce high titers of antigen-specific antibodies and a strong T-cell-dependent protective immune response in mice. The serum produced after immunization could also strongly bind to the surface of Helicobacter pylori NCTC 11637. Our research also found that a Helicobacter pylori glycoconjugate vaccine based on α-1,6-glucan in LPS could generate cross-reactive IgG antibodies in animals. Therefore, mannogen oligosaccharides and α-1,6-glucan may be important antigenic epitopes in Helicobacter pylori LPS.

[0005] In view of this, the present invention prepares three chimeric oligosaccharides based on the surface lipopolysaccharide of Helicobacter pylori, providing antigen candidates for the development of Helicobacter pylori vaccines and drugs. Summary of the Invention

[0006] The purpose of this invention is to provide three candidate antigen chimeric oligosaccharides of Helicobacter pylori and their preparation methods.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: The first aspect of the present invention provides three candidate antigen chimeric oligosaccharides of Helicobacter pylori, which are chimeric oligosaccharides derived from the lipopolysaccharide on the surface of Helicobacter pylori, specifically chimeric oligosaccharide I, chimeric oligosaccharide II, and chimeric oligosaccharide III, and their general chemical structural formulas are shown in formulas (I), (II), and (III) below, respectively: .

[0008] The second aspect of the present invention provides a method for preparing chimeric oligosaccharides of the three candidate antigens of Helicobacter pylori, comprising: (1) the preparation of chimeric oligosaccharide I, comprising the following steps: S1. Under the action of triethyl orthoacetate and p-toluenesulfonic acid monohydrate, the 2,3-hydroxyl groups in compound 1 with the structural formula (1) are protected by propylene oxide. Then, under the action of lithium tritert-butoxy aluminum hydride, the anodic position is reduced to a hydroxyl group. Then, under the action of dicyclohexylcarbodiimide and 4-dimethylaminopyridine, it is condensed with alkynyl ester to obtain D-glycerol-D-mannohepose alkynyl ester donor, namely compound 2 with the structural formula (2).

[0009] S2. The triethyl orthoacetate at the 2,3 position of compound 2 is ring-opened, and then the exposed hydroxyl group is protected by acetyl groups under the action of acetic anhydride and 4-dimethylaminopyridine to obtain compound 3 with the structural formula shown in (3). S3. Under the action of PPh3AuOTf and trifluoromethanesulfonic acid, compound 3 undergoes a glycosylation reaction with 1-azidopropanol to obtain compound 4 with the structural formula shown in (4).

[0010] S4. Compound 4 undergoes an azide reaction under the action of triphenylphosphine to reduce the azide group to an amino group, thereby obtaining compound 5 with the structural formula shown in (5). S5. Compound 6, with the structural formula shown in (6), reacts with o-nitrobenzyl bromide under the action of potassium hydroxide aqueous solution and tetrabutylammonium bromide, and then is protected with a photosensitive group ( o NB) protects the exposed 6-position hydroxyl group, and then, under the action of N-iodosuccinimide and boron trifluoride ether, it undergoes a glycosylation reaction with iodoacrylic acid. The resulting reaction product is then reacted with 1-hexyne under the action of bis(triphenylphosphine)palladium dichloride, cuprous iodide and triethylamine to obtain compound 7 with the structural formula shown in (7). S6. Compound 7 is dissolved in an organic solvent and undergoes a glycosylation reaction with the linker compound 8, which has the general structural formula shown in (8), under the action of SPhosAuNTf2 and N,N-dimethylformamide. After the reaction is complete, the photosensitive protecting group on the 6-position hydroxyl group is removed under light irradiation. o NB), to obtain compound 9 with the general structural formula as shown in (9); S7, compound 9 and compound 10 with the structural formula shown in (10) react under the action of sodium hydrogen to obtain compound 11 with the general structural formula shown in (11); S8. Dissolve compound 11 in a solvent and perform an azide reaction on compound 11 under the action of triphenylphosphine to reduce the azide group to an amino group, thereby obtaining compound 12 with the general structural formula as shown in (12). S9. Compound 12 and glutaric acid undergo a condensation reaction under the action of dicyclohexylcarbodiimide and 4-dimethylaminopyridine to obtain compound 13 with the general structural formula (13). S10. Under the action of 4-dimethylaminopyridine and carbodiimide, compound 5 and compound 13 undergo a condensation reaction to obtain compound 14 with the general structural formula (14). S11. Compound 14 is dissolved in a solvent, the propylene group is removed first, the acetyl group is removed under the action of sodium methoxide, and then the protecting group is removed by hydrogenation under the action of Pd(OH)2 / C to obtain chimeric oligosaccharide I. (2) Preparation of chimeric oligosaccharide II, including the following steps:

[0011] A1. Compound 9 was dissolved in an organic solvent. Under the action of SPhosAuNTf2 and N,N-dimethylformamide, compound 9 and compound 7 underwent a glycosylation reaction. After the reaction was complete, the photosensitive protecting group on the 6-hydroxyl group was removed by light irradiation. o NB), to obtain compound 16 with the general structural formula as shown in (16); A2. Under the action of sodium hydride, compound 16 reacts with compound 10 to obtain compound 17 with the general structural formula (17). A3. Dissolve compound 17 in a solvent and perform an azide reaction on compound 17 under the action of triphenylphosphine to reduce the azide group to an amino group, thereby obtaining compound 18 with the general structural formula (18). A4. Compound 18 undergoes a condensation reaction with glutaric acid under the action of dicyclohexylcarbodiimide and 4-dimethylaminopyridine to obtain compound 19 with the general structural formula (19). A5. Compound 5 and compound 19 undergo a condensation reaction under the action of 4-dimethylaminopyridine and carbodiimide to obtain compound 20 with the general structural formula (20). A6. Compound 20 was first depropylated, then deacetylated in the presence of sodium methoxide, and then dissolved in a solvent. The protecting group was removed by hydrogenation in the presence of Pd(OH)2 / C to obtain chimeric oligosaccharide II. (3) Preparation of chimeric oligosaccharide III, including the following steps:

[0012] B1. Compound 9 undergoes a glycosylation reaction with compound 7 under the action of SPhosAuNTf2 and N,N-dimethylformamide. After the reaction is complete, the photosensitive protecting group on the 6-hydroxyl group is removed by light irradiation. o NB), then compound 7 and SPhosAuNTf2 were added to the system again, and a glycosylation reaction occurred. After the reaction was complete, the system was irradiated with light to remove the photosensitive protecting group on the 6-hydroxyl group. o NB), to obtain compound 22 with the general structural formula as shown in (22); B2. Compound 22 reacts with compound 10 under the action of sodium hydrogen to obtain compound 23 with the general structural formula (23). B3. Compound 23 undergoes an azide reaction under the action of triphenylphosphine, reducing the azide group to an amino group to obtain compound 24 with the general structural formula (24). B4. Compound 24 is dissolved in a solvent, and under the action of dicyclohexylcarbodiimide and 4-dimethylaminopyridine, compound 24 undergoes a condensation reaction with glutaric acid to obtain compound 25 with the general structural formula (25). B5. Compound 5 and compound 25 undergo a condensation reaction under the action of 4-dimethylaminopyridine and carbodiimide to obtain compound 26 with the structural formula shown in (26). B6. Compound 26 was dissolved in a solvent, and the propylene group was removed first. Then, the acetyl group was removed under the action of sodium methoxide. Finally, the protecting group was removed by hydrogenation under the action of Pd(OH)2 / C to obtain the chimeric oligosaccharide III.

[0013] Preferably, in step S1, the molar ratio of compound 1, triethyl orthoacetate, p-toluenesulfonic acid monohydrate, lithium tri-tert-butoxyaluminum hydride, dicyclohexylcarbodiimide, 4-dimethylaminopyridine, and alkynyl olefinic acid is (1~1.2):(8~12):(0.1~0.3):(2~4):(1~2):(0.1~0.2):(1~2); the temperature for the propylene oxide protection reaction of the 2,3-hydroxy-orthoacetate is room temperature; the reaction temperature for the reduction of the anomeric position by lithium tri-tert-butoxyaluminum hydride is 0 °C; and the temperature for the condensation reaction with alkynyl olefinic acid is room temperature.

[0014] Preferably, in step S2, the molar ratio of compound 2, acetic anhydride, and 4-dimethylaminopyridine is 1:(8~12):(5~7); the ring-opening reaction of triethyl orthoacetate at the 2,3 position is at room temperature, and the acetyl protection reaction is at room temperature.

[0015] Preferably, in step S3, the molar ratio of compound 3, 1-azidopropanol, PPh3AuOTf, and trifluoromethanesulfonic acid is (1~1.2):(1.5~2):(0.2~0.6):(0.1~0.2); and the temperature of the glycosylation reaction is -40 °C.

[0016] Preferably, in step S4, the molar ratio of compound 4 to triphenylphosphine is (1~1.2):(4~6); and the temperature of the azidation reaction is 60 °C. Preferably, in step S5, the molar ratio of compound 6, tetrabutylammonium bromide, o-nitrobenzyl bromide, N-iodosuccinimide, boron trifluoride ether, iodoacrylic acid, palladium dichloride of bis(triphenylphosphine) chloride, cuprous iodide, 1-hexyne, and triethylamine is (1~1.2):(0.5~1):(4~5):(2~3):(2~3):(2~3):(0.05~0.1):(0.05~0.1):(1~2):(1~2); the reaction temperature in each step is room temperature. Preferably, in step S6, the molar ratio of compound 7, linker compound 8, SPhosAuNTf2, and N,N-dimethylformamide is (1~1.2):(1.5~2):(0.2~0.4):(4~6); the glycosylation reaction is carried out at 0 °C to room temperature; the organic solvent is a mixed solution of diethyl ether and dichloromethane, and the volume ratio of diethyl ether to dichloromethane is (3~4):1.

[0017] Preferably, in step S7, the molar ratio of compound 9, compound 10, and sodium hydride is (1~1.2):(1.5~2):(4~5); and the reaction temperature is room temperature to 80 °C. Preferably, in step S8, the molar ratio of compound 11 to triphenylphosphine is (1~1.2):(4~6); the temperature of the azidation reaction is 60 °C; the solvent is a mixed solution of tetrahydrofuran and water, and the volume ratio of tetrahydrofuran to water is (3~5):1. Preferably, in step S9, the molar ratio of compound 12, glutaric acid, dicyclohexylcarbodiimide, and 4-dimethylaminopyridine is (1~1.2):(8~10):(1.5~2):(0.4~0.5); and the temperature of the condensation reaction is room temperature.

[0018] Preferably, in step S10, the molar ratio of compound 5, compound 13, 4-dimethylaminopyridine, and carbodiimide is (1~1.2):(1~1.2):(0.4~0.5):(1.5~2); and the temperature of the condensation reaction is room temperature.

[0019] Preferably, in step S11, the solvent is a mixed solution of tert-butanol, water, and acetic acid, and the volume ratio of tert-butanol, water, and acetic acid is (3~4):1:(0.05~0.08). Preferably, in step A1, the molar ratio of compound 9, compound 7, SPhosAuNTf2 and N,N-dimethylformamide is (1~1.2):(1~1.2):(0.2~0.4):(4~6); the glycosylation reaction temperature is 0 °C to room temperature; the organic solvent is a mixed solution of diethyl ether and dichloromethane, and the volume ratio of diethyl ether to dichloromethane is (3~4):1.

[0020] Preferably, in step A2, the molar ratio of compound 16, compound 10, and sodium hydride is (1~1.2):(1.5~2):(4~5); and the reaction temperature is room temperature to 80 °C.

[0021] Preferably, in step A3, the molar ratio of compound 17 to triphenylphosphine is (1~1.2):(4~6); the temperature of the azidation reaction is 60 °C; the solvent is a mixed solution of tetrahydrofuran and water, and the volume ratio of tetrahydrofuran to water is (3~5):1.

[0022] Preferably, in step A4, the molar ratio of compound 18, glutaric acid, dicyclohexylcarbodiimide, and 4-dimethylaminopyridine is (1~1.2):(8~10):(1.5~2):(0.4~0.5); and the temperature of the condensation reaction is room temperature.

[0023] Preferably, in step A5, the molar ratio of compound 5, compound 19, 4-dimethylaminopyridine, and carbodiimide is (1~1.2):(1~1.2):(0.4~0.5):(1.5~2); and the temperature of the condensation reaction is room temperature.

[0024] Preferably, in step A6, the solvent is a mixed solution of tert-butanol, water, and acetic acid, and the volume ratio of tert-butanol, water, and acetic acid is (3~4):1:(0.05~0.08).

[0025] Preferably, in step B1, the molar ratio of compound 9, compound 7, SPhosAuNTf2 and N,N-dimethylformamide is (1~1.2):(1~1.2):(0.2~0.4):(4~6); the glycosylation reaction temperature is 0 °C to room temperature; the organic solvent is a mixed solution of diethyl ether and dichloromethane, and the volume ratio of diethyl ether to dichloromethane is (3~4):1.

[0026] Preferably, in step B2, the molar ratio of compound 22, compound 10, and sodium hydride is (1~1.2):(1.5~2):(4~5); and the reaction temperature is room temperature to 80 °C.

[0027] Preferably, in step B3, the molar ratio of compound 23 to triphenylphosphine is (1~1.2):(4~6); and the temperature of the azidation reaction is 60 °C.

[0028] Preferably, in step B4, the molar ratio of compound 24, glutaric acid, dicyclohexylcarbodiimide, and 4-dimethylaminopyridine is (1~1.2):(8~10):(1.5~2):(0.4~0.5); the solvent is a mixed solution of tetrahydrofuran and water, and the volume ratio of tetrahydrofuran to water is (3~5):1; the condensation reaction is carried out at room temperature.

[0029] Preferably, in step B5, the molar ratio of compound 5, compound 25, 4-dimethylaminopyridine, and carbodiimide is (1~1.2): (1~1.2): (0.4~0.5): (1.5~2); and the condensation reaction temperature is room temperature.

[0030] Preferably, in step B6, the solvent is a mixed solution of tert-butanol, water, and acetic acid, and the volume ratio of tert-butanol, water, and acetic acid is (3~4):1:(0.05~0.08).

[0031] The fifth aspect of the present invention provides the use of the above-mentioned chimeric oligosaccharide I, chimeric oligosaccharide II, and chimeric oligosaccharide III in the preparation of Helicobacter pylori vaccines or anti-Helicobacter pylori drugs.

[0032] Beneficial effects (1) This invention provides chimeric oligosaccharides of three candidate antigens of Helicobacter pylori and their preparation methods. These chimeric oligosaccharides are all derived from lipopolysaccharides on the surface of Helicobacter pylori. This invention starts with mannoheptose lactone compounds with exposed 2 and 3-hydroxyl groups, benzyl protection at 4, and hydroxypropylidene protection at 6 and 7. The 2 and 3-hydroxyl groups are protected by triethyl orthoacetate, the anolyte is reduced, an acetylacetate is added to the anolyte, the triethyl orthoacetate ring is opened, and the 3-hydroxyl group is protected by acetyl to prepare mannoheptose acetylacetate donors. Then, under the action of PPh3AuOTf and trifluoromethanesulfonic acid, it undergoes a glycosylation reaction with 1-azidopropanol, and the azide group is further reduced to an amino group. Starting from glucosinolates with exposed 6-hydroxyl groups and benzyl protection at 2, 3, and 4, the glycosides are photosensitized and protected by photosensitive protection. The 6-hydroxyl group was protected by a photosensitive protecting group, followed by iodoacrylate and coupling with 1-hexyne to obtain a glucosynyl ester donor protected by a photosensitive protecting group. A one-pot photosensitive process was then performed using SPhosAuNTf2 and N,N-dimethylformamide to prepare an α-1,6-linked oligosaccharide. The 6-hydroxyl group of the oligosaccharide was then protected with an azide group, and the azide group was further reduced to an amino group. Glutaric acid was used as a linker to connect the two fragments, and after three steps of deprotection (propionyl group removal, acetyl group removal, and hydrogenation), three chimeric oligosaccharides derived from Helicobacter pylori lipopolysaccharide were finally obtained. The chimeric oligosaccharides from Helicobacter pylori lipopolysaccharide provided by this invention have the potential to serve as antigen candidates for Helicobacter pylori glycosuriac vaccines and can be used in the preparation of Helicobacter pylori vaccines or anti-Helicobacter pylori drugs.

[0033] (2) The preparation method of the present invention is mild and simple to operate, and has potential application prospects for biocontrol agents. Attached Figure Description

[0034] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0035] Figure 1 The structural diagrams are of chimeric oligosaccharide I, chimeric oligosaccharide II, and chimeric oligosaccharide III; Figure 2 The synthetic route for chimeric oligosaccharide I, specifically compound 15; Figure 3 The synthetic route for chimeric oligosaccharide II, specifically compound 21; Figure 4 The synthetic route for chimeric oligosaccharide III, specifically compound 27. Detailed Implementation

[0036] In the following description, specific details such as particular system architectures and techniques are set forth for illustrative purposes and not for limitation, in order to provide a thorough understanding of the embodiments of the invention. However, those skilled in the art will understand that the invention may be implemented in other embodiments without these specific details.

[0037] Example 1 Reference Figure 2 The preparation of chimeric oligosaccharide I specifically involves compound 15 (m=5, R is -(CH2)5-NBnCbz, L is -(CH2)5-NH2). Specific steps: (1) Compound 1 (67.6 mg, 0.20 mmol) was dissolved in redistilled toluene (2 mL), and triethyl orthoacetate (0.4 mL, 2 mmol, 10.0 equiv) and p-toluenesulfonic acid monohydrate (8 mg, 0.04 mmol, 0.2 equiv) were added under argon atmosphere. After stirring at room temperature for half an hour, TLC monitoring showed that the reaction was complete. After quenching with saturated sodium bicarbonate solution, the upper toluene organic layer of the system was removed, and the lower aqueous phase was extracted three times with dichloromethane. The organic phases were combined and dried with anhydrous sodium sulfate to remove water. The crude product obtained by filtration and concentration was directly used in the next step.

[0038] The crude product from the previous step was dissolved in redistilled tetrahydrofuran (5.7 mL). After cooling the system to 0 °C, lithium aluminum tritert-butoxyhydride (102 mg, 0.4 mmol, 2.0 equiv) was added, and the reaction was maintained at 0 °C under argon atmosphere overnight. After TLC monitoring showed that the reaction was complete, the reaction was quenched with saturated sodium bicarbonate solution, and the system was extracted three times with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate to remove water, filtered, concentrated, and evaporated to dryness. The resulting crude product was used directly in the next step of the reaction.

[0039] The crude product from the previous step was dissolved in dry dichloromethane (4 mL). Under argon protection, 4-dimethylaminopyridine (3.8 mg, 0.03 mmol, 0.15 equiv) and dicyclohexylcarbodiimide (50 mg, 0.24 mmol, 1.2 equiv) were added dropwise to the system after being dissolved in dry dichloromethane (1 mL). The reaction was allowed to proceed at room temperature. After TLC monitoring showed complete reaction, the reaction was quenched with saturated sodium bicarbonate solution, extracted three times with dichloromethane, and the organic phases were combined. The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Separation was achieved by column chromatography (petroleum ether:ethyl acetate = 30:1 → 18:1) to give a pale yellow syrup compound 2 (67 mg, 62% over three steps). The characterization result was: [α]25D = +10.8 ( c 0.33, CHCl3); 1 H NMR (400 MHz, CDCl3) δ 7.37–7.31 (m, 5 H), 6.38(d, J = 4.0 Hz, 1 H), 6.23 (d, J = 11.2 Hz, 1 H), 5.98 (d, J = 11.2 Hz, 1 H),4.86–4.81 (m, 1 H), 4.63 (d, J = 11.2 Hz, 1 H), 4.55 (t, J = 6.4 Hz, 1 H), 4.33(dq, J = 3.6, 7.6 Hz, 2 H), 3.91 (dt, J = 2.4, 5.2 Hz, 2 H), 3.82 (dd, J = 6.8,8.4 Hz, 1 H), 3.60–3.54 (m, 2 H), 3.49 (dd, J= 6.0, 9.2 Hz, 1 H), 2.46–2.41(m, 2 H), 1.58–1.49 (m, 4 H), 1.28 (s, 3 H), 1.25 (s, 6 H), 1.19 (d, J = 7.2Hz, 3 H), 0.88 (d, J = 7.2 Hz, 3 H); 13 C NMR (150 MHz, CDCl3) δ 162.5, 138.0,128.4, 128.2, 126.1, 125.9, 109.9, 106.3, 90.8, 75.9, 75.1, 73.3, 71.3, 70.5,65.7, 58.6, 30.5, 29.8, 26.3, 25.3, 22.1, 21.1, 20.0, 18.6, 13.7; HRMS (ESI) m / z calcd for C 30 H 40 O9Na [M + Na] + 567.2570, found 567.2571. (2) Compound 2 (39 mg, 0.072 mmol) was dissolved in dry tetrahydrofuran (5 mL), and 80% aqueous acetic acid solution (5 mL) was added dropwise to the system. The mixture was stirred at room temperature for 3 hours. After the reaction was complete as indicated by TLC, the reaction was quenched with saturated sodium bicarbonate solution. The mixture was extracted three times with dichloromethane, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and evaporated to dryness. The product was then used directly in the next step. The product from the previous step was dissolved in dry dichloromethane (7 mL), and 4-dimethylaminopyridine (44 mg, 0.36 mmol, 5.0 equiv) was added. Then, acetic anhydride (68 mg, 0.072 mmol) was added dropwise to the system. μ L, 0.72 mmol, 10.0 equiv), reacted at room temperature for one hour under argon protection. After the reaction was complete, saturated sodium bicarbonate solution was added to quench the reaction. The mixture was extracted three times with dichloromethane, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated by column chromatography (petroleum ether:ethyl acetate = 15:1 → 5:1) to give a colorless, transparent oily compound 3 (27 mg, 67% over two steps). The characterization result was: [α]25 D = +92.0 ( c 0.39, CHCl3); 1 H NMR (400 MHz, CDCl3) δ 7.33–7.22 (m, 5 H), 6.24 (dt, J= 2.4, 11.2 Hz, 1 H), 6.15 (d, J = 2.4 Hz, 1 H), 5.97 (d, J = 11.6 Hz, 1 H), 5.41 (dd, J = 3.6, 9.2Hz, 1 H), 5.25 (dd, J = 2.4, 3.2 Hz, 1 H), 4.72–4.59 (m, 2 H), 4.37–4.28 (m, 1H), 4.01–3.93 (m, 2 H), 3.87 (dd, J = 7.2, 8.0 Hz, 1 H), 3.78 (t, J = 9.2 Hz, 1H), 2.39 (td, J = 2.4, 7.2 Hz, 2 H), 2.12 (s, 3 H), 1.97 (s, 3 H), 1.50–1.43(m, 2 H), 1.42 (s, 3 H), 1.30 (d, J = 10.4 Hz, 5 H), 0.82 (t, J = 7.2 Hz, 3 H); 13 C NMR (150 MHz, CDCl3) δ 169.8, 169.8, 162.2, 137.7, 128.6, 128.1, 128.1,126.5, 125.6, 109.9, 106.7, 90.6, 78.0, 75.8, 74.8, 74.2, 73.3, 71.5, 68.9,65.5, 30.4, 26.3, 25.1, 22.1, 20.9, 19.9, 13.7; HRMS (ESI) m / z calcd forC 30 H 38 O 10 Na [M + Na] + 581.2363, found 581.2362. (3) Compound 3 (60 mg, 0.11 mmol) was mixed with 1-azidopropanol (20 mg, 0.11 mmol) μ L, 0.22 mmol, 2.0 equiv) was dissolved in dry toluene (1 ml), and activated 4 Å molecular sieve (60 mg) was added. The mixture was stirred at room temperature for 15 min, then cooled to -40 °C. After holding at this temperature for 15 min, trifluoromethanesulfonic acid (2 L, 0.22 mmol, 2.0 equiv) was added under argon protection. μ L (0.022 mmol, 0.2 equiv) was reacted with freshly prepared gold catalyst PPh3AuOTf (0.1 M in CH2Cl2, 0.44 mL, 0.044 mmol, 0.4 equiv) and stirred overnight at -40 °C to room temperature. TLC monitoring showed complete conversion of the donor. The mixture was then neutralized to neutral by adding triethylamine, concentrated under vacuum, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10 / 1 → 4 / 1) to give a white syrupy product 4 (32.9 mg, 60%, α only). Characterization results showed: [α]25 D = +42.7 ( c 0.63, CHCl3); 1 HNMR (400 MHz, CDCl3) δ 7.40–7.27 (m, 5 H), 5.34 (dd, J = 3.6, 9.2 Hz, 1 H), 5.22 (dd, J = 2.0, 3.2 Hz, 1 H), 4.77 (d, J = 2.0 Hz, 1 H), 4.65 (q, J = 10.8Hz, 2 H), 4.38 (td, J = 2.8, 6.4 Hz, 1 H), 4.03 (dd, J = 6.4, 8.0 Hz, 1 H),3.89–3.75 (m, 3 H), 3.66 (t, J = 9.6 Hz, 1 H), 3.53–3.34 (m, 3 H), 2.13 (s, 3H), 1.99 (s, 3 H), 1.85 (q, J = 6.4 Hz, 2 H), 1.46 (s, 3 H), 1.34 (s, 3 H); 13 CNMR (150 MHz, CDCl3) δ 170.1, 169.9, 137.7, 128.6, 128.1, 109.6, 97.5, 75.5,74.6, 74.2, 72.0, 71.0, 70.1, 64.7, 64.7, 48.3, 28.8, 26.3, 25.0, 21.0, 21.0;HRMS (ESI) m / z calcd for C 24 H 33 N3O9Na [M + Na] +530.2115, found 530.2115. (4) Compound 4 (60 mg, 0.12 mmol) was dissolved in a mixed solvent of tetrahydrofuran and water (V / V = 4:1, 6.5 mL), and triphenylphosphine (0.186 g, 0.71 mmol, 6.0 equiv) was added. The mixture was stirred at 60 °C for 3 h. After the reaction of the starting material was complete as monitored by TLC, the system was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 30 / 1 → 10 / 1) to obtain a colorless syrupy product 5 (40 mg, 70%). The characterization results were as follows: 1 H NMR (400 MHz, CDCl3) δ7.34–7.29 (m, 5 H), 5.36–5.28 (m, 1 H), 5.22 (dd, J = 2.0, 3.6 Hz, 1 H), 4.79(d, J = 2.0 Hz, 1 H), 4.69–4.60 (m, 2 H), 4.41–4.33 (m, 1 H), 4.02 (dd, J =6.4, 8.0 Hz, 1 H), 3.92–3.85 (m, 1 H), 3.79 (tq, J = 3.2, 7.6 Hz, 2 H), 3.69–3.61 (m, 1 H), 3.50 (dt, J = 6.0, 10.0 Hz, 1 H), 2.91 (td, J = 1.6, 7.0 Hz, 2H), 2.12 (s, 3 H), 1.98 (s, 3 H), 1.84 (p, J = 6.4 Hz, 2 H), 1.46 (s, 4 H), 1.34 (s, 3 H). (5) Compound 6 (0.80 g, 1.3 mmol) was dissolved in dichloromethane (6.5 mL), tetrabutylammonium bromide (0.42 g, 1.0 mmol, 0.8 equiv) and 33% potassium hydroxide aqueous solution (3.2 mL) were added, and the mixture was stirred at room temperature for 10 min. o-Nitrobenzyl bromide (0.730 g, 3.4 mmol, 2.6 equiv) was added, and the mixture was stirred at room temperature for 30 min. 33% potassium hydroxide aqueous solution (3.2 mL) and o-nitrobenzyl bromide (0.730 g, 3.4 mmol, 2.6 equiv) were added, and the mixture was stirred at room temperature for 30 min. The reaction was monitored by TLC until it was complete. The reaction was quenched by adding water to the system. The aqueous phase was extracted three times with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under vacuum and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 30 / 1 → 20 / 1) to give a brown syrupy product (0.661 g, 68%).

[0040] The product from the previous step (53 mg, 0.060 mmol) and iodoacrylic acid (36 mg, 0.18 mmol, 3.0 equiv) were dissolved in dichloromethane (1.0 mL), and activated 4 Å molecular sieve (50 mg) was added. The mixture was stirred at room temperature for 15 min, and then N-iodosuccinimide (27 mg, 0.12 mmol, 2.0 equiv) and boron trifluoride diethyl ether (15 mL) were added under argon protection. μ The reaction was carried out at room temperature for 2 h with stirring (L, 0.12 mmol, 2.0 equiv). After the reaction was complete as monitored by TLC, the reaction was quenched by adding saturated sodium thiosulfate solution. The aqueous phase was extracted three times with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 12 / 1 → 8 / 1) to give a yellow slurry product (34 mg, 73%, α:β = 1:0.4).

[0041] The resulting yellow paste (1.43 g, 1.9 mmol) was dissolved in acetonitrile (37 mL), and palladium dichloride bis(triphenylphosphine) (65 mg, 0.093 mmol, 0.050 equiv) and cuprous iodide (35 mg, 0.19 mmol, 0.10 equiv) were added. The system was purged with argon five times, and under argon protection, triethylamine (0.34 mL, 3.7 mmol, 2.0 equiv) and 1-hexyne (0.32 mL, 2.8 mmol, 1.5 equiv) were added, and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until complete. A saturated ammonium chloride solution was added to quench the reaction. The aqueous phase was extracted three times with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under vacuum and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 12 / 1 → 8 / 1) to give a brown slurry product, compound 7 (1.20 g, 88%). Characterization results are as follows: 1 H NMR (400 MHz, CDCl3) δ8.07 (d, J = 8.0 Hz, 1 H), 7.81 (dd, J = 8.0, 16.4 Hz, 1 H), 7.62 (t, J = 7.6Hz, 1 H), 7.42 (t, J = 8.0 Hz, 1 H), 7.37–7.26 (m, 12 H), 7.23 (t, J = 2.8 Hz, 3 H), 6.54 (d, J = 3.6 Hz, 1 H), 6.26 (td, J = 3.6, 11.6 Hz, 1 H), 6.12–5.92(m, 1 H), 5.01–4.59 (m, 8 H), 4.31 (t, J = 6.8 Hz, 1 H), 4.11–3.96 (m, 1 H), 3.92–3.70 (m, 4 H), 2.45 (td, J = 2.8, 7.2 Hz, 2 H), 1.49 (dt, J = 7.2, 22.0Hz, 2 H), 1.39–1.24 (m, 2 H), 0.95–0.82 (m, 3 H); 13C NMR (150 MHz, CDCl3) δ163.2, 147.1, 138.7, 138.2, 137.8, 135.2, 133.9, 129.0, 128.8, 128.7, 128.6,128.6, 128.6, 128.5, 128.3, 128.2, 128.1, 128.1, 128.1, 128.0, 128.0, 128.0,127.9, 127.9, 127.9, 126.6, 125.6, 124.8, 105.9, 94.2, 90.2, 81.9, 79.2,75.9, 75.5, 73.2, 73.1, 70.0, 69.4, 65.7, 30.6, 22.1, 20.0, 13.7, 13.7; HRMS(ESI) m / z calcd forC 43 H 45 NO9Na [M + Na] + 742.2992 was found as 742.2991. (6) Transfer 6 digits o NB-protected monosaccharide donor 7 (40 mg, 0.054 mmol) and five-carbon linker 8 (35 mg, 0.11 mmol, 2.0 equiv) were dissolved in a mixed solvent of diethyl ether and dichloromethane (V / V = 3:1, 1.0 mL), and activated 4 Å molecular sieve (50 mg) and N,N-dimethylformamide (24 mmol) were added. μ L, 0.32 mmol, 6.0 equiv), stirred at room temperature for 15 min, then cooled to 0 ℃. After holding at this temperature for 15 min, the freshly prepared gold catalyst SPhosAuNTf2 (0.27 M in CH2Cl2, 0.10 mL, 0.027 mmol, 0.50 equiv) was added under argon protection, and the mixture was stirred at 0 ℃ to room temperature for 2 h. TLC monitoring showed that the reaction was complete. The system was then transferred to a parallel light reactor and stirred at room temperature for 5 min under 365 nm UV irradiation. TLC monitoring showed that the starting material reacted completely. The reaction solution was diluted and filtered, concentrated under vacuum, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 6 / 1 → 2 / 1) to obtain a colorless slurry product, compound 9 (23 mg, 55% over two steps, α only). Angew. Chem. Int. Ed. 2022, 61 The characterization of (e202202554) is performed, and the characterization results are as follows: 1H NMR (400 MHz, CDCl3) δ 7.44–7.12 (m, 26 H), 5.21–5.12 (m, 2 H), 4.98 (d, J = 10.8 Hz, 1 H), 4.91–4.72 (m, 3 H), 4.64 (h, J = 4.8 Hz, 3 H), 4.55–4.43 (m,2 H), 3.99 (t, J = 9.2 Hz, 1 H), 3.81–3.70 (m, 1 H), 3.70–3.46 (m, 5 H), 3.42–3.13 (m, 3 H), 1.61–1.47 (m, 4 H), 1.37 (s, 2 H). (7) Compound 9 (0.40 g, 0.53 mmol) was dissolved in dry tetrahydrofuran (5.0 mL), and 60% sodium hydride (0.11 g, 2.6 mmol, 5.0 equiv) was slowly added. After stirring at room temperature for 1.5 h, the temperature was raised to 80 °C and stirred for another 1.5 h. Then the temperature was lowered to 50 °C, and compound 10 (0.27 mg, 1.1 mmol, 2.0 equiv) was added under argon protection. The reaction was maintained at 50 °C and stirred for 12 h. After TLC monitoring showed that the starting material was completely converted, water was added to the system under ice bath to quench the reaction. The aqueous phase was extracted three times with dichloromethane, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and collected, concentrated under vacuum, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8 / 1 → 4 / 1) to obtain a colorless slurry product, namely compound 11 (0.33 g, 73%). The characterization result is: [α]25 D = +16.6 ( c 0.99, CHCl3); 1 H NMR (400MHz, CDCl3) δ 7.41–7.26 (m, 14 H), 7.26–7.08 (m, 11 H), 5.19–5.08 (d, J = 12.4Hz, 2 H), 4.95 (d, J = 10.8 Hz, 1 H), 4.86 (d, J = 10.8 Hz, 1 H), 4.80–4.70 (m,2 H), 4.67 (d, J = 5.2 Hz, 1 H), 4.62–4.52 (m, 2 H), 4.46 (d, J= 7.6 Hz, 2 H), 3.93 (t, J = 9.2 Hz, 1 H), 3.71–3.58 (m, 2 H), 3.57–3.45 (m, 5 H), 3.44–3.36 (m, 1 H), 3.36–3.26 (m, 3 H), 3.25–3.09 (m, 2 H), 1.87–1.72 (m, 2 H), 1.50(dd, J = 6.0, 10.8 Hz, 3 H), 1.28 (d, J = 16.0 Hz, 2 H); 13 C NMR (150 MHz, CDCl3)δ 138.9, 138.5, 138.4, 128.9, 128.9, 128.9, 128.8, 128.8, 128.7, 128.7,128.6, 128.5, 128.5, 128.5, 128.2, 128.2, 128.1, 128.1, 128.0, 128.0, 127.9,127.9, 127.8, 127.7, 127.7, 127.6, 127.4, 127.3, 127.3, 127.2, 127.2, 97.0,82.2, 80.2, 75.8, 73.3, 70.2, 69.7, 68.3, 67.3, 48.5, 29.3, 29.2, 29.2, 23.6;HRMS (ESI) m / z calcd for C 50 H 58 N4O8Na [M + Na] + 865.4152, found 865.4149. (8) Compound 11 (0.32 g, 0.38 mmol) was dissolved in a mixed solvent of tetrahydrofuran and water (V / V = 4:1, 13 mL), and triphenylphosphine (0.593 g, 2.3 mmol, 6.0 equiv) was added. The mixture was stirred at 60 °C for 3 h. After the reaction of the starting material was complete as monitored by TLC, the system was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 40 / 1 → 15 / 1) to obtain a colorless slurry product, namely compound 12 (0.24 mg, 77%). The characterization results are as follows: 1 H NMR (400 MHz, CDCl3) δ 7.35–7.02 (m, 26 H), 5.15–5.03 (m, 2 H), 4.88 (d,J = 10.8 Hz, 1 H), 4.78 (dd, J = 7.2, 10.8 Hz, 2 H), 4.70 (d, J = 11.2, 1 H), 4.65 (d, J = 12.0, 1H), 4.57 (d, J = 12.0 Hz, 1 H), 4.49 (d, J = 11.2 Hz, 1 H), 4.41 (d, J = 9.2 z,2 H), 3.87 (t, J = 9.2 Hz, 1 H), 3.64 (s, 1 H), 3.57–3.29 (m, 8 H), 3.22–3.05 (m, 2 H), 2.99 (q, J = 6.0 Hz, 2 H), 1.91–1.76 (m, 2 H), 1.60–1.40 (m, 4 H), 1.28–1.18 (m, 4 H). (9) Compound 12 (0.29 g, 0.35 mmol) and glutaric acid (0.457 g, 3.5 mmol, 10 equiv) were dissolved in acetonitrile (15 mL), and 4-dimethylaminopyridine (21 mg, 0.17 mmol, 0.50 equiv) and dicyclohexylcarbodiimide (0.143 g, 0.69 mmol, 2.0 equiv) were added. The mixture was stirred at room temperature for 2 h. After the reaction of the starting materials was complete by TLC monitoring, saturated sodium bicarbonate solution was added to the system to quench the reaction. The aqueous phase was extracted three times with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, and the filtrate was collected by filtration. The filtrate was concentrated under vacuum and purified by silica gel column chromatography (dichloromethane:methanol = 20 / 1 → 10 / 1) to obtain a colorless slurry product, namely compound 13 (0.28 g, 86%). The characterization results are as follows: 1 H NMR (400MHz, CDCl3) δ 7.47–7.26 (m, 21 H), 7.26–7.10 (m, 4 H), 6.63–6.43 (m, 1 H), 5.17 (d, J = 13.2 Hz, 2 H), 5.00 (d, J = 10.8 Hz, 1 H), 4.91–4.74 (m, 3 H), 4.69 (m, 2 H), 4.57 (d, J= 11.2 Hz, 1 H), 4.49 (d, J = 6.0 Hz, 2 H), 3.99 (t, J = 9.2 Hz, 1 H), 3.70 (s, 1 H), 3.66–3.58 (m, 2 H), 3.56–3.44 (m, 5 H), 3.39(q, J = 5.6, 6.6 Hz, 2 H), 3.25 (s, 3 H), 2.33 (d, J = 7.2 Hz, 2 H), 2.18–2.04(m, 2 H), 1.90 (q, J = 6.8, 8.0 Hz, 2 H), 1.72 (q, J = 4.4, 4.8 Hz, 2 H), 1.63–1.48 (m, 4 H), 1.36–1.29 (m, 2 H). (10) Compound 5 (29 mg, 0.060 mmol) and compound 13 (61 mg, 0.066 mmol, 1.1 equiv) were dissolved in dichloromethane (1.0 mL), and 4-dimethylaminopyridine (3.7 mg, 0.030 mmol, 0.5 equiv) and carbodiimide (23 mg, 0.12 mmol, 2.0 equiv) were added. The mixture was stirred at room temperature for 3 h. After the reaction was complete as monitored by TLC, saturated sodium bicarbonate solution was added to quench the reaction. The aqueous phase was extracted three times with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The filtrate was purified by silica gel column chromatography (dichloromethane:methanol = 30 / 1 → 10 / 1) to obtain a colorless slurry product, namely compound 14 (65 mg, 77%). The characterization result was: [α]25 D = +20.6 ( c 0.32, CHCl3); 1 H NMR (400 MHz, CDCl3) δ 7.32–7.04 (m, 30 H), 6.21 (d, J =37.6 Hz, 2 H), 5.23 (dd, J = 3.6, 9.6 Hz, 1 H), 5.19–5.13 (m, 1 H), 5.09 (d, J = 12.0 Hz, 2 H), 4.90 (d, J = 10.8 Hz, 1 H), 4.80 (d, J= 10.8 Hz, 1 H), 4.73(d, J = 10.8 Hz, 1 H), 4.70–4.59 (m, 3 H), 4.59–4.46 (m, 4 H), 4.41 (d, J = 5.6Hz, 2 H), 4.30 (td, J = 2.8, 6.4 Hz, 1H), 3.98–3.84 (m, 2 H), 3.80–3.76 (dd, J = 2.8, 9.6 Hz, 1 H), 3.74–3.59 (m, 3 H), 3.58–3.52 (m, 2 H), 3.52–3.31 (m, 8H), 3.21 (m, 6 H), 2.11 (q, J = 7.6 Hz, 3 H), 2.05 (d, J = 3.6 Hz, 3 H), 2.01(s, 1 H), 1.91 (s, 3 H), 1.80 (q, J = 6.8 Hz, 3 H), 1.74–1.59 (m, 5 H), 1.57–1.43 (m, 4 H), 1.36 (d, J = 12.0 Hz, 3 H), 1.26 (s, 3 H); 13 C NMR (150 MHz,CDCl3) δ 172.8, 172.6, 170.1, 169.9, 138.3, 137.6, 128.6, 128.6, 128.6,128.6, 128.5, 128.5, 128.4, 128.4, 128.1, 128.1, 128.0, 128.0, 127.9, 127.9,127.9, 127.8, 127.6, 109.5, 97.3, 97.0, 77.4, 76.9, 76.9, 75.7, 75.3, 74.6,73.9, 73.1, 72.1, 70.8, 70.1, 70.0, 66.1, 64.4, 35.5, 35.4, 34.0, 29.2, 29.1,26.2, 25.7, 25.0, 25.0, 22.1, 21.0; HRMS (ESI) m / z calcd for C 79 H 99 N3O 19 Na [M +Na]+ 1416.6770, found 1416.6766. (11) Compound 14 (50 mg, 0.036 mmol) was dissolved in 80% aqueous acetic acid solution (4.5 mL) and stirred at 70 °C for 2 h. TLC monitoring showed that the reaction was complete, and the crude product was obtained by concentrating the system and used directly in subsequent steps.

[0042] The crude product obtained in the previous step was dissolved in methanol (1.0 mL), and sodium methoxide (0.78 mg, 0.014 mmol, 0.4 equiv) was added. The mixture was stirred at room temperature for 30 min. TLC monitoring showed that the reaction of the starting material was complete. Hydrogen ion exchange resin was added to the system to neutralize it. The filtrate was collected by filtration, concentrated under vacuum, and purified by silica gel column chromatography (dichloromethane:methanol = 12 / 1 → 10 / 1) to obtain a white slurry product.

[0043] The white paste-like product obtained in the previous step was dissolved in a mixed solvent of tert-butanol and water (V / V = 4:1, 3.3 mL), and 10% Pd(OH)2 / C (90 mg) and acetic acid (52 mg) were added. μ The system was replaced with hydrogen five times, and the reaction was stirred at room temperature for 12 h. TLC monitoring showed that the reaction was complete. After filtration to remove the catalyst, the filtrate was concentrated under vacuum and purified by LH-20 dextran gel column chromatography (MeOH) to obtain a colorless slurry product, compound 15 (13 mg, 52% over three steps). Characterization results showed: [α]25 D = +5.7 ( c 0.32, MeOH); 1 H NMR (400 MHz, D2O) δ 4.91 (d, J = 3.6 Hz, 1H), 4.83 (d, J = 1.6 Hz, 1 H), 4.04 (dt, J = 3.2, 7.6 Hz, 1 H), 3.93 (d, J = 2.4Hz, 1 H), 3.86–3.66 (m, 9 H), 3.63–3.51 (m, 4 H), 3.50–3.33 (m, 3 H), 3.32–3.23 (m, 3 H), 3.02 (t, J = 7.6 Hz, 1 H), 2.27 (t, J = 7.6 Hz, 3 H), 2.12 (dd, J= 10.8, 20.0 Hz, 1 H), 1.92–1.75 (m, 6 H), 1.74–1.65 (m, 3 H), 1.53–1.38 (m, 3 H), 1.34–1.20 (m, 3 H); 13 C NMR (150 MHz, D2O) δ 175.7, 175.7, 99.8, 98.0,73.1, 73.0, 71.6, 71.2, 70.9, 70.5, 69.9, 69.7, 69.2, 68.7, 67.8, 67.4, 65.1,61.7, 39.3, 36.4, 36.3, 34.9, 34.9, 28.2, 28.1, 28.0, 26.5, 22.8, 22.5, 21.8;HRMS (ESI) m / z calcd for C 29 H 55 N3O 15 H [M + H] + 686.3711, found 686.3712. Example 2 Reference Figure 3 The preparation of chimeric oligosaccharide II specifically involves compound 21 (m=5, R is -(CH2)5-NBnCbz, L is -(CH2)5-NH2). Specific steps: (1) Convert 6 bits o NB-protected monosaccharide donor 7 (68 mg, 0.095 mmol) and monosaccharide acceptor 9 (60 mg, 0.079 mmol, 0.4 equiv) were dissolved in a mixed solvent of diethyl ether and dichloromethane (V / V = 3:1, 1.0 mL), and activated 4 Å molecular sieve (50 mg) and N,N-dimethylformamide (37 mg) were added. μL, 0.047 mmol, 6.0 equiv), stirred at room temperature for 15 min, then cooled to 0 °C. After holding at this temperature for 15 min, freshly prepared gold catalyst SPhosAuNTf2 (0.40 M in CH2Cl2, 0.10 mL, 0.040 mmol, 0.5 equiv) was added under argon protection, and the mixture was stirred at 0 °C to room temperature for 2 h. After TLC monitoring showed complete acceptor conversion, the system was transferred to a parallel light reactor and stirred at room temperature for 10 min under 365 nm UV irradiation. TLC monitoring showed complete reaction of the starting material. The reaction solution was diluted and filtered, concentrated under vacuum, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 6 / 1 → 2 / 1) to obtain a colorless slurry product, compound 16 (50 mg, 53% over two steps, α only). Angew. Chem. Int. Ed. 2022, 61 The characterization of (e202202554) is performed, and the characterization results are as follows: 1 H NMR (400 MHz, CDCl3)δ 7.38–7.25 (m, 29 H), 7.25–7.10 (m,11 H), 5.16 (d, J = 8.8 Hz, 2 H), 4.95 (dd, J = 4.4, 11.6 Hz, 3 H), 4.92–4.85(m, 2 H), 4.79 (dd, J = 9.2, 10.8 Hz, 2 H), 4.68 (d, J = 4.8 Hz, 1 H), 4.67–4.62 (m, 4 H), 4.60 (d, J = 3.6 Hz, 1 H), 4.49 (m, 3 H), 3.97 (t, J = 9.2 Hz, 2H), 3.83 (dd, J = 3.6, 11.6 Hz, 1 H), 3.78–3.56 (m, 7 H), 3.53–3.45 (m, 2 H), 3.41 (dd, J = 3.6, 9.6 Hz, 1 H), 3.35–3.13 (m, 3 H), 1.48 (m, 4 H), 1.36–1.26 (m, 2 H). (2) Compound 16 (0.25 g, 0.21 mmol) was dissolved in dry tetrahydrofuran (3.0 mL), and 60% sodium hydride (42 mg, 1.1 mmol, 5.0 equiv) was slowly added. After stirring at room temperature for 1.5 h, the temperature was raised to 80 °C and stirred for another 1.5 h. Then the temperature was lowered to 50 °C, and compound 10 (0.11 g, 0.42 mmol, 2.0 equiv) was added under argon protection. The reaction was maintained at 50 °C and stirred for 12 h. After TLC monitoring showed that the starting material was completely converted, water was added to the system under ice bath to quench the reaction. The aqueous phase was extracted three times with dichloromethane, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and collected, concentrated under vacuum, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8 / 1 → 3 / 1) to obtain a white slurry product, namely compound 17 (0.18 g, 68%). The characterization result is: [α]25 D = +52.7 ( c 0.42, CHCl3); 1 H NMR (400MHz, CDCl3) δ 7.39–7.21 (m, 17 H), 7.21–7.01 (m, 23 H), 5.08 (d, J = 9.2 Hz, 2H), 4.99–4.79 (m, 5 H), 4.79–4.61 (m, 3 H), 4.60 (s, 1 H), 4.58–4.52 (m, 3H), 4.49 (d, J = 11.2 Hz, 1 H), 4.45 (d, J = 11.6 Hz, 1 H), 4.40 (d, J = 7.6Hz, 2 H), 3.89 (td, J = 6.0, 9.2 Hz, 2 H), 3.77 (dd, J = 3.6, 12.0 Hz, 1 H), 3.71–3.55 (m, 4 H), 3.50 (tt, J = 4.8, 9.6 Hz, 3 H), 3.43 (dt, J = 3.2, 9.6 Hz,3 H), 3.33 (m, 2 H), 3.28–3.19 (m, 3 H), 3.15 (t, J = 7.2 Hz, 1 H), 3.10 (t, J = 7.6 Hz, 1 H), 1.72 (tt, J= 6.8, 13.2 Hz, 2 H), 1.55–1.37 (m, 4 H), 1.21(dt, J = 7.0, 16.0 Hz, 2 H); 13 C NMR (150 MHz, CDCl3) δ 139.0, 138.8, 138.7,138.6, 138.5, 128.6, 128.5, 128.5, 128.5, 128.2, 128.0, 128.0, 127.9, 127.9,127.8, 127.7, 127.7, 97.3, 96.8, 82.2, 81.8, 80.5, 80.1, 77.8, 77.7, 75.7,75.7, 75.0, 73.2, 72.3, 70.7, 70.3, 69.6, 68.2, 67.2, 65.9, 48.5, 29.2, 23.6; HRMS (ESI) m / z calcd for C 77 H 86 N4O 13 Na [M + Na] + 1297.6089, found 1297.6091. (3) Compound 17 (137 mg, 0.11 mmol) was dissolved in a mixed solvent of tetrahydrofuran and water (V / V = 4:1, 3.2 mL), and triphenylphosphine (0.169 g, 0.64 mmol, 6.0 equiv) was added. The mixture was stirred at 60 °C for 3 h. After the reaction of the starting material was complete as monitored by TLC, the system was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 40 / 1 → 8 / 1) to obtain a colorless slurry product, namely compound 18 (114 mg, 85%). The characterization results are as follows: 1 H NMR (400 MHz, CDCl3) δ 7.44–7.03 (m, 40 H), 5.12 (d, J = 10.8 Hz, 2 H), 5.01–4.81 (m, 5 H), 4.79–4.58 (m, 7 H), 4.52 (dd, J = 6.8, 11.6 Hz, 2 H), 4.47–4.37 (m, 2 H), 3.91(dt, J1.50 (s, 4 H), 1.30–1.18 (m, 2 H). (4) Compound 18 (103 mg, 0.083 mmol) and glutaric acid (0.11 g, 0.83 mmol, 10.0 equiv) were dissolved in acetonitrile (3.0 mL), and 4-dimethylaminopyridine (5.0 mg, 0.041 mmol, 0.5 equiv) and carbodiimide (33 mg, 0.17 mmol, 2.0 equiv) were added. The mixture was stirred at room temperature for 2 h. After the reaction of the starting materials was complete by TLC monitoring, saturated sodium bicarbonate solution was added to the system to quench the reaction. The aqueous phase was extracted three times with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, and the filtrate was collected by filtration. The filtrate was concentrated under vacuum and purified by silica gel column chromatography (dichloromethane:methanol = 20 / 1 → 12 / 1) to obtain a colorless slurry product, namely compound 19 (83 mg, 74%). The characterization results are as follows: 1 H NMR (400MHz, CDCl3) δ 7.37–7.26 (m, 25 H), 7.26–7.11 (m, 15 H), 6.51 (s, 1 H), 5.15(s, 2 H), 5.03–4.77 (m, 7 H), 4.75–4.58 (m, 5 H), 4.58–4.42 (m, 4 H), 3.97(t, J = 8.8 Hz, 2 H), 3.85 (d, J = 11.6 Hz, 1 H), 3.76 (s, 2 H), 3.70 (q, J =7.2, 11.6 Hz, 2 H), 3.63–3.40 (m, 9 H), 3.31 (s, 1 H), 3.30–3.08 (m, 4 H), 2.32 (t, J = 6.8 Hz, 2 H), 2.08 (dq, J = 7.2, 13.6 Hz, 2 H), 1.93–1.85 (m, 2H), 1.70 (s, 2 H), 1.54 (s, 4 H), 1.33 (d, J = 6.6 Hz, 2 H). (5) Compound 5 (22 mg, 0.045 mmol) and compound 19 (68 mg, 0.050 mmol, 1.1 equiv) were dissolved in dichloromethane (1.0 mL), and 4-dimethylaminopyridine (2.8 mg, 0.023 mmol, 0.50 equiv) and carbodiimide (17 mg, 0.090 mmol, 2.0 equiv) were added. The mixture was stirred at room temperature for 3 h. After TLC monitoring showed that the reaction was complete, saturated sodium bicarbonate solution was added to quench the reaction. The aqueous phase was extracted three times with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum and purified by silica gel column chromatography (dichloromethane:methanol = 50 / 1 → 30 / 1) to obtain a colorless slurry product, namely compound 20 (60 mg, 72%). The characterization result was: [α]25 D = +93.9 ( c 0.13, CHCl3); 1 H NMR (400 MHz, CDCl3) δ 7.24 (dt, J = 5.6, 10.4 Hz, 20H), 7.20–7.03 (m, 25 H), 6.07 (dt, J = 6.0, 17.2 Hz, 2 H), 5.24 (dd, J = 3.2, 9.2 Hz, 1 H), 5.15 (t, J = 2.8 Hz, 1 H), 5.08 (d, J = 9.6 Hz, 2 H), 4.92–4.78(m, 5 H), 4.74–4.67 (m, 3 H), 4.63–4.55 (m, 6 H), 4.53–4.46 (m, 2 H), 4.43–4.37 (m, 2 H), 4.30 (td, J = 2.8, 6.8 Hz, 1 H), 3.99–3.84 (m, 3 H), 3.81–3.63(m, 7 H), 3.62–3.46 (m, 6 H), 3.43–3.29 (m, 7 H), 3.20 (m, 6 H), 2.11 (q, J =7.2 Hz, 3 H), 2.05 (s, 3 H), 1.91 (s, 3 H), 1.80 (q, J= 7.2 Hz, 2 H), 1.72–1.60 (m, 5 H), 1.44 (m, 4 H), 1.38 (s, 3 H), 1.27 (s, 3 H), 1.21 (s, 2 H); 13 CNMR (100 MHz, CDCl3) δ 172.8, 172.6, 170.1, 169.9, 139.1, 138.9, 138.6,138.6, 138.5, 138.1, 137.7, 128.7, 128.5, 128.5, 128.5, 128.2, 128.2, 128.1,128.1, 128.1, 128.0, 128.0, 127.9, 127.9, 127.8, 127.8, 127.7, 127.7, 127.4,109.6, 97.4, 96.8, 82.2, 81.8, 80.5, 80.2, 77.8, 77.6, 77.5, 75.7, 75.6,75.4, 75.0, 74.7, 74.0, 73.2, 72.3, 72.2, 70.9, 70.7, 70.2, 70.1, 70.0, 69.4,67.3, 66.2, 66.0, 64.5, 37.7, 37.2, 35.6, 35.4, 29.3, 29.2, 26.3, 25.1, 23.6,22.2, 21.0; HRMS (ESI) m / z calcd for C 106 H 127 N3O 24 Na [M + Na] + 1849.8741, found 1849.8732. (6) Compound 20 (22 mg, 0.012 mmol) was dissolved in 2.1 mL of 80% acetic acid aqueous solution and stirred at 70 °C for 2 h. TLC monitoring showed that the reaction was complete. The crude product was obtained by concentrating the system and used directly in subsequent steps.

[0044] The crude product obtained in the previous step was dissolved in methanol (1.0 mL), and sodium methoxide (0.26 mg, 0.0048 mmol, 0.4 equiv) was added. The mixture was stirred at room temperature for 30 min. TLC monitoring showed that the reaction of the starting material was complete. Hydrogen ion exchange resin was added to the system to neutralize it. The filtrate was collected by filtration, concentrated under vacuum, and purified by silica gel column chromatography (dichloromethane:methanol = 14 / 1 → 10 / 1) to obtain a yellow slurry product.

[0045] The product obtained in the previous step was dissolved in a mixed solvent of tert-butanol and water (V / V = 4:1, 3.4 mL), and 10% Pd(OH)2 / C (60 mg) and acetic acid (52 mg) were added. μ The system was replaced with hydrogen five times and stirred at room temperature for 24 h. TLC monitoring showed that the reaction was complete. After filtration to remove the catalyst, the filtrate was concentrated under vacuum and purified by LH-20 dextran gel column chromatography (MeOH) to obtain a colorless slurry product, compound 21 (5.0 mg, 49% over three steps). Characterization results showed: [α]25 D = +25.4 ( c 0.20, MeOH); 1 H NMR (400 MHz, D2O) δ 4.85 (t, J = 3.2 Hz, 3 H), 3.95 (dd, J = 3.6, 7.2 Hz, 1 H), 3.92–3.83 (m, 3 H), 3.79–3.74 (m, 2 H), 3.73 (d, J = 3.6 Hz, 1 H), 3.70 (d, J = 6.4 Hz, 2 H), 3.66 (d, J = 3.2 Hz, 2 H), 3.65–3.60(m, 3 H), 3.60–3.52 (m, 3 H), 3.52–3.41 (m, 6 H), 3.40–3.34 (m, 2 H), 3.20(p, J = 6.8 Hz, 4 H), 2.97–2.91 (m, 2 H), 2.19 (t, J = 7.6 Hz, 4 H), 1.82–1.69(m, 6 H), 1.63 (dt, J = 7.6, 15.2 Hz, 4H), 1.44–1.37 (m, 2H); 13C NMR (150 MHz, D2O) δ 175.7, 99.8, 98.1, 97.8, 73.4, 73.0, 71.6, 71.6, 71.2, 70.9, 70.4,70.2, 69.9, 69.7, 69.5, 69.1, 68.6, 67.9, 67.4, 67.2, 65.1, 61.7, 39.3, 36.4,36.3, 34.9, 28.2, 28.1, 28.0, 26.5, 23.2, 22.4, 21.8; HRMS (ESI) m / z calcd forC 35 H 65 N3O 20 H [M + H] + 848.4240, found 848.4238. Example 3 Reference Figure 4 The preparation of chimeric oligosaccharide III specifically involves compound 27 (m=5, R is -(CH2)5-NBnCbz, L is -(CH2)5-NH2). Specific steps: (1) Convert 6 bits o NB-protected monosaccharide donor 7 (36 mg, 0.051 mmol, 1.2 equiv) and monosaccharide acceptor 9 (32 mg, 0.042 mmol) were dissolved in a mixed solvent of diethyl ether and dichloromethane (V / V = 3:1, 1.0 mL), and activated 4 Å molecular sieve (50 mg) and N,N-dimethylformamide (19 mg) were added. μ L, 0.25 mmol, 6.0 equiv), stirred at room temperature for 15 min, then cooled to 0 °C. After holding at this temperature for 15 min, the freshly prepared gold catalyst SPhosAuNTf2 (0.21 M in CH2Cl2, 0.10 mL, 0.021 mmol, 0.5 equiv) was added under argon protection, and the mixture was stirred at 0 °C to room temperature for 2 h. After TLC monitoring showed that the reaction was complete, the system was transferred to a parallel light reactor and stirred at room temperature for 5 min under 365 nm UV irradiation. After TLC monitoring showed that the starting material reaction was complete, monosaccharide donor 7 (72 mg, 0.10 mmol, 2.4 equiv) and N,N-dimethylformamide (19 μL, 0.25 mmol, 6.0 equiv), under argon protection at 0 ℃, added freshly prepared gold catalyst SPhosAuNTf2 (0.21 M in CH2Cl2, 0.10 mL, 0.021 mmol, 0.5 equiv), and stirred at 0 ℃ to room temperature for 2 h. After TLC monitoring showed complete acceptor reaction, the system was transferred to a parallel light reactor and stirred at room temperature for 10 min under 365 nm UV irradiation. TLC monitoring showed complete reaction of the starting material, the reaction solution was diluted and filtered, concentrated under vacuum, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 8 / 1 → 2 / 1) to obtain a white slurry product, compound 22 (20 mg, 30% over four steps, α only). Angew. Chem. Int. Ed. 2022, 61 The characterization of (e202202554) is performed, and the characterization results are as follows: 1 H NMR (400 MHz, CDCl3) δ 7.39–7.26 (m, 32 H),7.26–7.09 (m, 23 H), 5.15 (d, J = 9.2 Hz, 2 H), 4.99 (d, J = 3.6 Hz, 1 H),4.98–4.84 (m, 7 H), 4.81–4.73 (m, 3 H), 4.63 (m, 7 H), 4.57–4.43 (m, 5 H), 4.04–3.67 (m, 11 H), 3.66–3.53 (m, 5 H), 3.51–3.43 (m, 2 H), 3.40 (dt, J =2.8, 9.6 Hz, 2 H), 3.33–3.10 (m, 3 H), 1.44 (d, J = 10.0 Hz, 4 H), 1.33–1.29(m, 2 H). (2) Compound 22 (176 mg, 0.11 mmol) was dissolved in dry tetrahydrofuran (3.0 mL), and 60% sodium hydride (21 mg, 0.53 mmol, 5.0 equiv) was slowly added. After stirring at room temperature for 1.5 h, the temperature was raised to 80 °C and stirred for another 1.5 h. Then the temperature was lowered to 50 °C, and compound 10 (54 mg, 0.21 mmol, 2.0 equiv) was added under argon protection. The reaction was stirred at 50 °C for 12 h. After TLC monitoring showed that the reaction was complete, water was added to the system under ice bath to quench the reaction. The aqueous phase was extracted three times with dichloromethane, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected. The filtrate was concentrated under vacuum and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8 / 1 → 4 / 1) to obtain a white slurry product, namely compound 23 (122 mg, 66%). The characterization result is: [α]25 D = +54.5 ( c 0.17, CHCl3); 1 H NMR (400 MHz, CDCl3) δ 7.38–7.26 (m, 30 H), 7.26–7.09 (m, 25 H), 5.16 (d, J = 8.0 Hz, 2 H), 5.04 (d, J = 3.6 Hz, 1 H), 4.98–4.85 (m, 7 H), 4.81–4.73 (m, 3 H), 4.68–4.44 (m, 12 H), 4.01–3.89 (m, 3 H), 3.86–3.63 (m, 9 H), 3.62–3.44 (m, 7 H), 3.43–3.26 (m, 6 H), 3.26–3.13 (m, 2 H), 1.86–1.72 (m, 2 H), 1.49 (s, 2 H), 1.34–1.22 (m, 4 H); 13C NMR (150 MHz, CDCl3) δ 139.1, 138.8, 138.7, 128.7, 128.6,128.6, 128.5, 128.5, 128.4, 128.2, 128.2, 128.1, 128.0, 128.0, 127.8, 127.7,127.7, 127.6, 97.4, 96.9, 82.2, 81.8, 81.7, 80.5, 80.2, 80.2, 77.7, 75.7,75.6, 75.1, 73.3, 72.3, 72.2, 70.7, 70.3, 69.6, 68.2, 67.3, 65.9, 65.8, 48.5,31.6, 30.3, 29.8, 29.3, 29.2, 23.6; HRMS (ESI) m / z calcd for C 104 H 114 N4O 18 Na [M +Na] + 1730.8059, found 1730.8076. (3) Compound 23 (74 mg, 0.043 mmol) was dissolved in a mixed solvent of tetrahydrofuran and water (V / V = 4:1, 1.9 mL), and triphenylphosphine (68 mg, 0.26 mmol, 6.0 equiv) was added. The mixture was stirred at 60 °C for 3 h. After the reaction of the starting material was complete as monitored by TLC, the system was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 25 / 1 → 15 / 1) to obtain a white slurry product, namely compound 24 (58 mg, 80%). The characterization results are as follows: 1 H NMR (400 MHz, CDCl3) δ 7.29 (dt, J = 9.9, 3.3 Hz, 22 H), 7.26–7.11 (m, 33 H), 5.14 (d, J =7.6 Hz, 2 H), 5.06–4.95 (m, 2 H), 4.94–4.86 (m, 5 H), 4.85–4.77 (m, 2 H), 4.75–4.68 (m, 3 H), 4.65 (dd, J = 4.0, 12.8 Hz, 4 H), 4.61–4.53 (m, 3 H), 4.45(s, 4 H), 3.92 (p, J= 9.2, 10.2 Hz, 4 H), 3.79 (t, J = 10.4 Hz, 3 H), 3.73 (d, J = 10.4 Hz, 3 H), 3.67 (d, J = 12.0 Hz, 2 H), 3.61–3.49 (m, 3 H), 3.47–3.34(m, 7 H), 3.31–3.04 (m, 4 H), 2.95–2.86 (m, 1 H), 2.78 (s, 1 H), 1.86–1.63(m, 4 H), 1.34–1.20 (m, 4H). (4) Compound 24 (58 mg, 0.034 mmol) and glutaric acid (45 mg, 0.34 mmol, 10.0 equiv) were dissolved in acetonitrile (2.0 mL), and 4-dimethylaminopyridine (2.1 mg, 0.017 mmol, 0.5 equiv) and carbodiimide (13 mg, 0.068 mmol, 2.0 equiv) were added. The mixture was stirred at room temperature for 2 h. After the reaction of the starting materials was complete by TLC monitoring, saturated sodium bicarbonate solution was added to quench the reaction. The aqueous phase was extracted three times with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The filtrate was then purified by silica gel column chromatography (dichloromethane:methanol = 20 / 1 → 10 / 1) to obtain a colorless slurry product, namely compound 25 (54 mg, 87%). The characterization results were as follows: 1 H NMR (400MHz, CDCl3) δ 7.40–7.26 (m, 32 H), 7.26–7.11 (m, 23 H), 6.45 (t, J = 5.2 Hz, 1H), 5.16 (d, J = 8.4 Hz, 2 H), 5.03 (d, J = 3.6 Hz, 1 H), 4.99–4.84 (m, 7 H), 4.83–4.70 (m, 4 H), 4.69–4.51 (m, 9 H), 4.50–4.41 (m, 3 H), 3.97 (td, J = 6.0,9.2 Hz, 3 H), 3.90–3.69 (m, 7 H), 3.68–3.52 (m, 6 H), 3.51–3.38 (m, 7 H), 3.26–3.13 (m, 3 H), 2.30 (dd, J= 6.0, 7.6 Hz, 2 H), 2.14–2.01 (m, 2 H), 1.89(q, J = 6.8 Hz, 2 H), 1.69 (h, J = 4.4, 4.8 Hz, 2 H), 1.54 (s, 3 H), 1.34–1.23 (m, 3 H). (5) Compound 5 (16 mg, 0.032 mmol) and compound 25 (64 mg, 0.035 mmol, 1.1 equiv) were dissolved in dichloromethane (1.0 mL), and 4-dimethylaminopyridine (2.0 mg, 0.016 mmol, 0.5 equiv) and carbodiimide (14 mg, 0.064 mmol, 2.0 equiv) were added. The mixture was stirred at room temperature for 3 h. After TLC monitoring showed that the reaction was complete, saturated sodium bicarbonate solution was added to quench the reaction. The aqueous phase was extracted three times with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum and purified by silica gel column chromatography (dichloromethane:methanol = 50 / 1 → 25 / 1) to obtain a colorless slurry product, namely compound 26 (56 mg, 75%). The characterization result was: [α]25 D = +20.6 ( c 0.40, CHCl3); 1 H NMR (400 MHz, CDCl3) δ 7.38–7.26 (m, 35 H), 7.26–7.11 (m, 25 H), 6.15 (t, J = 5.6 Hz, 2 H), 5.31 (dd, J = 3.2, 9.2 Hz, 1 H), 5.22(dd, J = 2.0, 3.2 Hz, 1 H), 5.16 (d, J = 8.4 Hz, 2 H), 5.01 (d, J = 3.6 Hz, 1H), 4.97–4.92 (m, 4 H), 4.91–4.84 (m, 2 H), 4.82–4.72 (m, 4 H), 4.65 (qd, J =6.0, 11.0 Hz, 9 H), 4.57–4.50 (m, 3 H), 4.46 (d, J = 8.4 Hz, 2 H), 4.38 (td, J = 3.2, 6.8 Hz, 1 H), 4.02 (dd,J = 6.0, 8.0 Hz, 1 H), 3.95 (m, 3 H), 3.88–3.83(m, 2 H), 3.76 (dq, J = 7.6, 12.0 Hz, 6 H), 3.65 (qd, J = 4.8, 12.0 Hz, 5 H),3.59–3.51 (m, 3 H), 3.51–3.33 (m, 9 H), 3.32–3.19 (m, 5 H), 3.15 (s, 1 H),2.22–2.16 (m, 2 H), 2.14 (d, J = 5.2 Hz, 4 H), 2.10 (d, J = 7.4 Hz, 1 H), 1.99(s, 3 H), 1.86 (p, J = 7.2 Hz, 2 H), 1.78 (d, J = 2.4 Hz, 1 H), 1.73 (s, 5 H),1.71–1.67 (m, 2 H), 1.46 (s, 3 H), 1.35 (d, J = 4.4 Hz, 3 H), 1.28 (d, J = 11.6Hz, 2 H); 13C NMR (150 MHz, CDCl3) δ 172.8, 172.7, 170.1, 170.0, 139.1, 139.0,138.8, 138.6, 138.5, 137.7, 128.7, 128.5, 128.5, 128.5, 128.4, 128.4, 128.2,128.2, 128.1, 128.0, 128.0, 127.9, 127.9, 127.8, 127.8, 127.7, 127.6, 127.5,109.6, 97.4, 97.3, 96.8, 82.2, 81.8, 81.7, 80.5, 80.5, 80.2, 77.8, 77.6, 77.5, 75.7, 75.6, 75.6, 75.4, 75.0, 75.0, 74.7, 74.0, 73.2, 72.4, 72.2, 72.2, 70.9, 70.8, 70.2, 70.0, 70.0, 69.3, 68.0, 67.2, 66.2, 65.9, 64.5, 37.6, 37.1, 35.5, 35.4, 29.3, 29.2, 29.2, 26.3, 25.1, 23.6, 22.2, 21.0; HRMS (ESI) m / z calcd for C 133 H 155 N3O 29 H [M + H] + 2260.0858, found 2260.0588. (6) Compound 26 (33 mg, 0.015 mmol) was dissolved in 3.2 mL of 80% acetic acid aqueous solution and stirred at 70 °C for 2 h. TLC monitoring showed that the reaction was complete. The crude product was obtained by concentrating the system and used directly in subsequent steps.

[0046] The crude product obtained in the previous step was dissolved in methanol (1.0 mL), and sodium methoxide (0.32 mg, 0.0060 mmol, 0.4 equiv) was added. The mixture was stirred at room temperature for 30 min. TLC monitoring showed that the reaction of the starting material was complete. Hydrogen ion exchange resin was added to the system to neutralize it. The filtrate was collected by filtration, concentrated under vacuum, and purified by silica gel column chromatography (dichloromethane:methanol = 12 / 1 → 8 / 1) to obtain a white slurry product.

[0047] The product obtained in the previous step was dissolved in a mixed solvent of tert-butanol and water (V / V = 4:1, 1.5 mL), and 10% Pd(OH)2 / C (50 mg) and acetic acid (25 mL) were added. μ The system was purged with hydrogen five times and stirred at room temperature for 36 h. TLC monitoring showed that the reaction was complete. The catalyst was removed by filtration, and the filtrate was concentrated under vacuum and purified by LH-20 dextran gel column chromatography (MeOH) to obtain a colorless slurry product, compound 27 (8.0 mg, 54% over three steps). Characterization results showed: [α]25 D = +38.0 ( c 0.22, MeOH); 1 H NMR (400 MHz, D2O) δ 4.95 (dd, J = 4.0, 7.6 Hz, 3 H), 4.83 (s, 1 H), 4.07–3.95 (m, 3 H), 3.93 (s, 2 H), 3.88–3.83 (m, 2 H), 3.81(d, J = 3.6 Hz, 1 H), 3.79–3.66 (m, 13 H), 3.65–3.56 (m, 5 H), 3.53 (d, J =10.0 Hz, 4 H), 3.44 (t, J = 10.0 Hz, 1 H), 3.36–3.25 (m, 4 H), 2.95 (t, J = 7.6Hz, 2 H), 2.27 (t, J = 7.6 Hz, 4 H), 1.93–1.79 (m, 6 H), 1.66 (q, J = 7.2 Hz, 4H), 1.47 (dt, J = 7.2, 14.8 Hz, 2 H); 13C NMR (150 MHz, D2O) δ 175.7, 162.9,99.8, 98.1, 97.8, 97.7, 73.4, 73.4, 73.0, 73.0, 71.6, 71.4, 71.3, 71.2, 70.9,70.4, 70.2, 70.1, 69.9, 69.7, 69.5, 69.5, 69.1, 68.6, 68.0, 67.4, 65.6, 65.1,61.7, 39.5, 36.4, 36.3, 28.2, 28.1, 27.3, 22.5, 21.8; HRMS (ESI) m / z calcd forC 41 H 75 N3O 25 Na [M + Na] + 1322.4587, found 1322.4590. This invention is not limited to the specific embodiments described above. Any modifications made by those skilled in the art based on the above concept without creative effort are within the scope of protection of this invention.

Claims

1. A chimeric oligosaccharide containing three candidate antigens of Helicobacter pylori, characterized in that, The candidate antigen chimeric oligosaccharides are chimeric oligosaccharide I, chimeric oligosaccharide II, and chimeric oligosaccharide III, with the following general chemical structural formulas (I), (II), and (III), respectively: 。 2. The method for preparing chimeric oligosaccharides containing three candidate antigens of Helicobacter pylori as described in claim 1, characterized in that: include: (1) Preparation of chimeric oligosaccharide I, including the following steps: S1. Under the action of triethyl orthoacetate and p-toluenesulfonic acid monohydrate, the 2,3-hydroxyl groups in compound 1 with the structural formula (1) are protected by propylene oxide. Then, under the action of lithium tritert-butoxy aluminum hydride, the anodic position is reduced to a hydroxyl group. Then, under the action of dicyclohexylcarbodiimide and 4-dimethylaminopyridine, it is condensed with alkynyl ester to obtain D-glycerol-D-mannohepose alkynyl ester donor, namely compound 2 with the structural formula (2). S2. The triethyl orthoacetate at the 2,3 position of compound 2 is ring-opened, and then the exposed hydroxyl group is protected by acetyl groups under the action of acetic anhydride and 4-dimethylaminopyridine to obtain compound 3 with the structural formula shown in (3). S3. Under the action of PPh3AuOTf and trifluoromethanesulfonic acid, compound 3 undergoes a glycosylation reaction with 1-azidopropanol to obtain compound 4 with the structural formula shown in (4). S4. Compound 4 undergoes an azide reaction under the action of triphenylphosphine to reduce the azide group to an amino group, thereby obtaining compound 5 with the structural formula shown in (5). S5. Compound 6, with the structural formula shown in (6), reacts with o-nitrobenzyl bromide under the action of potassium hydroxide aqueous solution and tetrabutylammonium bromide, and then is protected with a photosensitive group ( o NB) protects the exposed 6-position hydroxyl group, and then, under the action of N-iodosuccinimide and boron trifluoride ether, it undergoes a glycosylation reaction with iodoacrylic acid. The resulting reaction product is then reacted with 1-hexyne under the action of bis(triphenylphosphine)palladium dichloride, cuprous iodide and triethylamine to obtain compound 7 with the structural formula shown in (7). S6. Compound 7 is dissolved in an organic solvent and undergoes a glycosylation reaction with the linker compound 8, which has the general structural formula shown in (8), under the action of SPhosAuNTf2 and N,N-dimethylformamide. After the reaction is complete, the photosensitive protecting group on the 6-position hydroxyl group is removed under light irradiation. o NB), to obtain compound 9 with the general structural formula as shown in (9); S7, compound 9 and compound 10 with the structural formula shown in (10) react under the action of sodium hydrogen to obtain compound 11 with the general structural formula shown in (11); S8. Dissolve compound 11 in a solvent and perform an azide reaction on compound 11 under the action of triphenylphosphine to reduce the azide group to an amino group, thereby obtaining compound 12 with the general structural formula as shown in (12). S9. Compound 12 and glutaric acid undergo a condensation reaction under the action of dicyclohexylcarbodiimide and 4-dimethylaminopyridine to obtain compound 13 with the general structural formula (13). S10. Under the action of 4-dimethylaminopyridine and carbodiimide, compound 5 and compound 13 undergo a condensation reaction to obtain compound 14 with the general structural formula (14). S11. Compound 14 is dissolved in a solvent, the propylene group is removed first, the acetyl group is removed under the action of sodium methoxide, and then the protecting group is removed by hydrogenation under the action of Pd(OH)2 / C to obtain chimeric oligosaccharide I. (2) Preparation of chimeric oligosaccharide II, including the following steps: A1. Compound 9 was dissolved in an organic solvent. Under the action of SPhosAuNTf2 and N,N-dimethylformamide, compound 9 and compound 7 underwent a glycosylation reaction. After the reaction was complete, the photosensitive protecting group on the 6-hydroxyl group was removed by light irradiation. o NB), to obtain compound 16 with the general structural formula as shown in (16); A2. Under the action of sodium hydride, compound 16 reacts with compound 10 to obtain compound 17 with the general structural formula (17). A3. Dissolve compound 17 in a solvent and perform an azide reaction under the action of triphenylphosphine to reduce the azide group to an amino group, thereby obtaining compound 18 with the general structural formula (18). A4. Compound 18 undergoes a condensation reaction with glutaric acid under the action of dicyclohexylcarbodiimide and 4-dimethylaminopyridine to obtain compound 19 with the general structural formula (19). A5. Compound 5 and compound 19 undergo a condensation reaction under the action of 4-dimethylaminopyridine and carbodiimide to obtain compound 20 with the general structural formula (20). A6. Compound 20 was first depropylated, then deacetylated in the presence of sodium methoxide, and then dissolved in a solvent. The protecting group was removed by hydrogenation in the presence of Pd(OH)2 / C to obtain chimeric oligosaccharide II. (3) The preparation of the chimeric oligosaccharide III includes the following steps: B1. Compound 9 undergoes a glycosylation reaction with compound 7 under the action of SPhosAuNTf2 and N,N-dimethylformamide. After the reaction is complete, the photosensitive protecting group on the 6-hydroxyl group is removed by light irradiation. o NB), then compound 7 and SPhosAuNTf2 were added to the system again, and a glycosylation reaction occurred. After the reaction was complete, the system was irradiated with light to remove the photosensitive protecting group on the 6-hydroxyl group. o NB), to obtain compound 22 with the general structural formula as shown in (22); B2. Compound 22 reacts with compound 10 under the action of sodium hydrogen to obtain compound 23 with the general structural formula (23). B3. Compound 23 undergoes an azide reaction under the action of triphenylphosphine, reducing the azide group to an amino group to obtain compound 24 with the general structural formula (24). B4. Compound 24 is dissolved in a solvent, and under the action of dicyclohexylcarbodiimide and 4-dimethylaminopyridine, compound 24 undergoes a condensation reaction with glutaric acid to obtain compound 25 with the general structural formula (25). B5. Compound 5 and compound 25 undergo a condensation reaction under the action of 4-dimethylaminopyridine and carbodiimide to obtain compound 26 with the structural formula shown in (26). B6. Compound 26 was dissolved in a solvent, and the propylene group was removed first. Then, the acetyl group was removed under the action of sodium methoxide. Finally, the protecting group was removed by hydrogenation under the action of Pd(OH)2 / C to obtain the chimeric oligosaccharide III.

3. The method for preparing chimeric oligosaccharides containing three candidate antigens of Helicobacter pylori according to claim 2, characterized in that, In step S1, the molar ratio of compound 1, triethyl orthoacetate, p-toluenesulfonic acid monohydrate, lithium tri-tert-butoxyaluminum hydride, dicyclohexylcarbodiimide, 4-dimethylaminopyridine, and alkynyl alkenoic acid is (1~1.2):(8~12):(0.1~0.3):(2~4):(1~2):(0.1~0.2):(1~2); the temperature for the propylene oxide protection reaction of the 2,3-hydroxy-orthoacetate is room temperature; the reaction temperature for the reduction of the anomeric position by lithium tri-tert-butoxyaluminum hydride is 0 °C; and the temperature for the condensation reaction with alkynyl alkenoic acid is room temperature.

4. The method for preparing chimeric oligosaccharides containing three candidate antigens of Helicobacter pylori according to claim 2, characterized in that: In step S2, the molar ratio of compound 2, acetic anhydride, and 4-dimethylaminopyridine is 1:(8~12):(5~7); the ring-opening reaction of triethyl orthoacetate at the 2,3 position is at room temperature, and the acetyl protection reaction is at room temperature. In step S3, the molar ratio of compound 3, 1-azidopropanol, PPh3AuOTf, and trifluoromethanesulfonic acid is (1~1.2):(1.5~2):(0.2~0.6):(0.1~0.2); the glycosylation reaction is carried out at a temperature of -40 °C. In step S4, the molar ratio of compound 4 to triphenylphosphine is (1~1.2):(4~6); the reaction temperature for reducing the azide group to an amino group is 60 °C. The reaction temperature in each step S5 is room temperature. The molar ratio of compound 6, tetrabutylammonium bromide, o-nitrobenzyl bromide, N-iodosuccinimide, boron trifluoride ether, iodoacrylic acid, palladium dichloride of bis(triphenylphosphine), cuprous iodide, 1-hexyne and triethylamine is (1~1.2):(0.5~1):(4~5):(2~3):(2~3):(2~3):(0.05~0.1):(0.05~0.1):(1~2):(1~2).

5. The method for preparing chimeric oligosaccharides containing three candidate antigens of Helicobacter pylori according to claim 2, characterized in that: In step S6, the molar ratio of compound 7, linker compound 8, SPhosAuNTf2, and N,N-dimethylformamide is (1~1.2):(1.5~2):(0.2~0.4):(4~6); the glycosylation reaction is carried out at 0 °C to room temperature; the organic solvent is a mixed solution of diethyl ether and dichloromethane, and the volume ratio of diethyl ether to dichloromethane is (3~4):

1. In step S7, the molar ratio of compound 9, compound 10, and sodium hydride is (1~1.2):(1.5~2):(4~5); the reaction temperature is from room temperature to 80 °C. In step S8, the molar ratio of compound 11 to triphenylphosphine is (1~1.2):(4~6); the reaction temperature for reducing the azide group to amino is 60 °C; the solvent is a mixed solution of tetrahydrofuran and water, and the volume ratio of tetrahydrofuran to water is (3~5):

1. In step S9, the molar ratio of compound 12, glutaric acid, dicyclohexylcarbodiimide, and 4-dimethylaminopyridine is (1~1.2):(8~10):(1.5~2):(0.4~0.5); the condensation reaction is carried out at room temperature. In step S10, the molar ratio of compound 5, compound 13, 4-dimethylaminopyridine, and carbodiimide is (1~1.2):(1~1.2):(0.4~0.5):(1.5~2); the condensation reaction is carried out at room temperature. In step S11, the solvent is a mixed solution of tert-butanol, water, and acetic acid, and the volume ratio of tert-butanol, water, and acetic acid is (3~4):1:(0.05~0.08).

6. The method for preparing chimeric oligosaccharides containing three candidate antigens of Helicobacter pylori according to claim 2, characterized in that: In step A1, the molar ratio of compound 9, compound 7, SPhosAuNTf2, and N,N-dimethylformamide is (1~1.2):(1~1.2):(0.2~0.4):(4~6); the glycosylation reaction is carried out at 0 °C to room temperature; the concentration of the solution obtained by dissolving compound 9 in an organic solvent is 0.05~0.1 mmol / mL; the organic solvent is a mixed solution of diethyl ether and dichloromethane, and the volume ratio of diethyl ether to dichloromethane is (3~4):1; In step A2, the molar ratio of compound 16, compound 10, and sodium hydride is (1~1.2):(1.5~2):(4~5); the reaction temperature is from room temperature to 80 °C. In step A3, the molar ratio of compound 17 to triphenylphosphine is (1~1.2):(4~6); the temperature of the azidation reaction is 60 °C; the solvent is a mixed solution of tetrahydrofuran and water, and the volume ratio of tetrahydrofuran to water is (3~5):

1.

7. The method for preparing chimeric oligosaccharides of three candidate antigens of Helicobacter pylori according to claim 2, characterized in that: In step A4, the molar ratio of compound 18, glutaric acid, dicyclohexylcarbodiimide, and 4-dimethylaminopyridine is (1~1.2):(8~10):(1.5~2):(0.4~0.5); the condensation reaction temperature is room temperature. In step A5, the molar ratio of compound 5, compound 19, 4-dimethylaminopyridine, and carbodiimide is (1~1.2):(1~1.2):(0.4~0.5):(1.5~2); the condensation reaction temperature is room temperature. In step A6, the solvent is a mixed solution of tert-butanol, water, and acetic acid, and the volume ratio of tert-butanol, water, and acetic acid is (3~4):1:(0.05~0.08).

8. The method for preparing chimeric oligosaccharides of three candidate antigens of Helicobacter pylori according to claim 2, characterized in that: In step B1, the molar ratio of compound 9, compound 7, SPhosAuNTf2, and N,N-dimethylformamide is (1~1.2):(1~1.2):(0.2~0.4):(4~6); the glycosylation reaction temperature is 0 °C to room temperature; In step B2, the molar ratio of compound 22, compound 10, and sodium hydride is (1~1.2):(1.5~2):(4~5); the reaction temperature is room temperature to 80 °C.

9. The method for preparing chimeric oligosaccharides of three candidate antigens of Helicobacter pylori according to claim 2, characterized in that: In step B3, the molar ratio of compound 23 to triphenylphosphine is (1~1.2):(4~6); the temperature of the azidation reaction is 60 °C. In step B4, the molar ratio of compound 24, glutaric acid, dicyclohexylcarbodiimide, and 4-dimethylaminopyridine is (1~1.2):(8~10):(1.5~2):(0.4~0.5); the solvent is a mixed solution of tetrahydrofuran and water, and the volume ratio of tetrahydrofuran to water is (3~5):1; the condensation reaction is carried out at room temperature. In step B5, the molar ratio of compound 5, compound 25, 4-dimethylaminopyridine, and carbodiimide is (1~1.2):(1~1.2):(0.4~0.5):(1.5~2); the condensation reaction temperature is room temperature. In step B6, the solvent is a mixed solution of tert-butanol, water, and acetic acid, and the volume ratio of tert-butanol, water, and acetic acid is (3~4):1:(0.05~0.08).

10. The use of chimeric oligosaccharides of the three candidate antigens of Helicobacter pylori as described in claim 1, specifically chimeric oligosaccharide I, chimeric oligosaccharide II, and chimeric oligosaccharide III, in the preparation of Helicobacter pylori vaccines or anti-Helicobacter pylori drugs.

Images