A T N Method for preparing antigen

Abstract

The present invention belongs to the field of biomedical technology and specifically relates to a T N The invention uses 2-deoxy-2-azidogalactosyl N-phenylacetimidate (PTFAI) as a donor and alcohol as an acceptor, and adopts fusion reagent regulation and remote participation effect to synthesize T with high stereoselectivity. N Antigens are used, avoiding the use of precious metal activators (AgClO4 or Ni(4‑F‑PhCN)4(OTf)2, etc.) and strong bases (P4‑t‑Bu, KH, etc.). The experimental operation is simple, the reaction system is mild, no other side reactions occur, the reaction is efficient and stereoselective, and the raw material price is low.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology and specifically relates to a T N Methods for preparing antigens. Background Art

[0002] Protein glycosylation can generally be divided into two categories: N-linked and O-linked. In N-linked glycoproteins, the N-acetylglucosamine unit (GlcNAc) is linked to the N of the asparagine amino acid side chain by a β-glycosidic bond. In O-linked glycoproteins, the N-acetylgalactosamine (GalNAc) unit is linked to the hydroxyl group of serine or threonine by an α-glycosidic bond, generating a core structure usually called T N antigen:

[0003]

[0004] The C(3)-OH or C(6)-OH of this core structure can be substituted to form a variety of structural motifs. These antigens are widely distributed in cell surface mucin glycoproteins and participate in cell adhesion events related to cancer metastasis, especially T N antigen, which is highly expressed in most epithelial cancers such as lung cancer, colon cancer, gastric cancer and breast cancer. N Antigens as biomarkers and therapeutic targets for tumor vaccine therapy.

[0005] When developing cancer vaccines, we need T cells with clear structures and high purity. N However, obtaining sufficient quantities of T cells in homogeneous form from natural sources is difficult. N Antigens are challenging. In many cases, high purity T N Antigens can only be obtained through chemical or enzymatic synthesis.

[0006] Chemical synthesis of T N Antigens often involve glycosidic linkages, and regio- and stereo-selectivity issues are the main challenges in glycosidic bond formation. In the past few decades, great progress has been made in the synthesis of complex oligosaccharides. However, the stereo-selective construction of 1,2-cis glycosidic bonds remains one of the most difficult problems in chemical synthesis.

[0007] Therefore, finding a highly stereoselective synthesis method has become a technical problem that needs to be solved urgently in this field. Summary of the invention

[0008] In view of this, the purpose of the present invention is to provide a preparation method with simple experimental operation, mild reaction system, no other side reactions, high reaction efficiency, high stereoselectivity and low raw material price to address the technical problems of low stereoselectivity, high raw material price and harsh reaction conditions in the prior art.

[0009] In order to achieve the above object, the present invention adopts the following technical solution:

[0010] A T N The preparation method of the antigen is to use 2-deoxy-2-azidogalactosyl PTFAI as a donor and alcohol as an acceptor, and dissolve them together with triphenylphosphine oxide in dry dichloromethane under argon protection, and then add freshly dried The molecular sieves were stirred for 15 minutes, and then iodotrimethylsilane (TMSI) was added dropwise to react at room temperature to complete the glycosidation reaction.

[0011] Considering T N The N-acetylgalactosamine (GalNAc) unit in the antigen structure is connected to the hydroxyl group of serine or threonine by a 1,2-cis glycosidic bond. The present invention uses 2-deoxy-2-azidogalactosyl PTFAI as a donor and alcohol as an acceptor, and adopts TMSI and Ph3PO fusion reagent regulation and acyl remote participation effect to synthesize T with high stereoselectivity. N Antigen, avoiding the use of precious metal activators (AgClO4 or Ni(4-F-PhCN)4(OTf)2, etc.) and strong bases (P4-t-Bu, KH, etc.). The experimental operation is simple, the reaction system is mild, no other side reactions occur, the reaction is efficient and stereoselective, and the raw material price is low.

[0012] Furthermore, the preparation method of the glycosylated product is:

[0013]

[0014] Under argon protection, 2-deoxy-2-azidogalactosyl PTFAI donor was dissolved in dry dichloromethane, freshly dried 3A molecular sieves and triphenylphosphine oxide were added in sequence, and after stirring at room temperature for 15 minutes, iodotrimethylsilane was added, and the mixture was stirred at room temperature for 1 hour. HOR was added, and the mixture was moved into a 30°C oil bath for reaction. After the reaction was monitored by thin layer chromatography, saturated sodium thiosulfate was added to quench the reaction, and the mixture was extracted three times with dichloromethane. The obtained organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated with a rotary evaporator, and purified by silica gel column chromatography to obtain a glycosylated product; wherein,

[0015] R 3 For B Z , Lev, Ac and other acyl functional groups;

[0016] R 4 For B Z , Lev, Ac and other acyl functional groups;

[0017] R 6It is an ether or ester group such as Bn, Lev, TBDPS, Ac, Bz, etc.

[0018] Furthermore, the preparation method of the glycosylated product is:

[0019]

[0020] Under argon protection, 2-deoxy-2-azidogalactosyl PTFAI donor 2 was dissolved in dry dichloromethane, and freshly dried 3A molecular sieves and triphenylphosphine oxide were added in sequence. After stirring at room temperature for 15 minutes, iodotrimethylsilane was added, and the mixture was stirred at room temperature for 1 hour. N-benzyloxycarbonyl-L-serine benzyl ester was added, and the mixture was moved into a 30°C oil bath for reaction for 5.5 days. Saturated sodium thiosulfate was added to quench the reaction, and the mixture was extracted three times with dichloromethane. The obtained organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated using a rotary evaporator. The mixture was purified by silica gel column chromatography to obtain a white solid 3.

[0021] Furthermore, the T N The method for preparing the antigen also includes group treatment, and the group treatment method is:

[0022]

[0023] Under argon protection, the white solid 3 was dissolved with pyridine, and then thioacetic acid was added dropwise. The reaction was allowed to react overnight at room temperature. The reaction was complete after monitoring by thin layer chromatography. The reaction solution was concentrated by rotary evaporator and purified by silica gel column chromatography to obtain intermediate 3-a.

[0024] The intermediate 3-a was dissolved in methanol, and then 0.6 mL of water, 0.06 mL of glacial acetic acid, and 10% palladium on carbon were added in sequence. Hydrogen was introduced into the reaction system and reacted at room temperature for 6 h. The reaction was completed after monitoring by thin layer chromatography. The reaction solution was filtered through a 25 mm diameter polyethersulfone microporous filter membrane, and the filtrate was concentrated by a rotary evaporator to obtain a white solid 3-b.

[0025] Under argon protection, the white solid 3-b was dissolved in methanol, and then 1,4-dioxane and sodium hydroxide aqueous solution were added in sequence, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored to be complete by thin layer chromatography. The reaction solution was adjusted to pH=7 with glacial acetic acid, and the reaction solution was concentrated by rotary evaporator and purified by gel column chromatography to obtain a white solid 4, which was T N antigen.

[0026] In the present invention, the T N The method for preparing the antigen also includes the method for preparing the galactosyl PTFAI donor:

[0027]

[0028] Under argon protection, 2-deoxy-2-azido-3,4,6-tri-O-benzoyl-α / β-D-pyranose galactose hemiacetal 1 and 2,2,2-trifluoro-N-phenylacetylimidoyl chloride were dissolved in acetone, and K2CO3 was added. The reaction was allowed to proceed overnight at room temperature, and the reaction was monitored to be complete by thin layer chromatography. The reaction was filtered through diatomaceous earth, and the filtrate was concentrated by rotary evaporator. The white solid donor 2 obtained by silica gel column chromatography was the donor 2-deoxy-2-azidogalactosyl PTFAI.

[0029] In some embodiments, the T disclosed in the present invention N The method for preparing the antigen includes:

[0030]

[0031] Preparation of donor: Under argon protection, 2-deoxy-2-azido-3,4,6-tri-O-benzoyl-α / β-D-pyranose galactose hemiacetal 1 (592.8 mg, 1.15 mmol) and 2,2,2-trifluoro-N-phenylacetyl imide chloride (261.6 mg, 1.26 mmol) were dissolved in 11 mL of acetone, and K2CO3 (238.3 mg, 1.72 mmol) was added. The reaction was allowed to proceed overnight at room temperature, and the reaction was monitored to be complete by thin layer chromatography. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated by rotary evaporator. The white solid 2 (774.5 mg) was obtained by silica gel column chromatography (petroleum ether + 2% triethylamine), with a yield of 98%.

[0032] Glycosidation reaction: Under argon protection, the above white solid 2 (574.9 mg, 0.83 mmol) was dissolved in 8.3 mL of dry dichloromethane, and freshly dried 3A molecular sieves (1 g) and triphenylphosphine oxide (1.39 g, 5.00 mmol) were added in sequence. After stirring at room temperature for 15 min, iodotrimethylsilane (0.12 mL, 0.83 mmol) was added, and the mixture was stirred at room temperature for 1 h. N-benzyloxycarbonyl-L-serine benzyl ester (409.1 mg, 1.25 mmol) was added, and the mixture was moved into a 30°C oil bath for reaction for 5.5 days. Saturated sodium thiosulfate was added to quench the reaction, and the mixture was extracted three times with dichloromethane. The obtained organic phase was washed with saturated sodium chloride in sequence, and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated with a rotary evaporator. Silica gel column chromatography (ethyl acetate / petroleum ether = 1:4) gave a white solid 3 (353.5 mg) with a yield of 51%.

[0033] Group treatment: Under argon protection, the above white solid 3 was dissolved with 7.6 mL pyridine, and then thioacetic acid (5.8 mL, 77.84 mmol) was added dropwise, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored to be complete by thin layer chromatography. The reaction solution was concentrated by rotary evaporator, and the intermediate (293.1 mg) was obtained by silica gel column chromatography (ethyl acetate: petroleum ether = 1:2-1:1), with a yield of 89%. The above intermediate (140 mg, 0.17 mmol) was dissolved with 6 mL methanol, and then 0.6 mL water, 0.06 mL glacial acetic acid, and 10% palladium carbon (352.7 mg, 0.33 mmol) were added in sequence. Hydrogen was introduced into the reaction system, and the reaction was allowed to proceed for 6 h at room temperature. The reaction was monitored to be complete by thin layer chromatography. The reaction solution was filtered with a 25 mm diameter polyethersulfone microporous filter membrane, and the filtrate was concentrated by rotary evaporator to obtain 84.8 mg white solid, with a yield of 83%. Under argon protection, the above white solid was dissolved in 1.5 mL of methanol, and then 1.5 mL of 1,4-dioxane and 2 mL of 1M sodium hydroxide aqueous solution were added in sequence, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored to be complete by thin layer chromatography. The reaction solution was adjusted to pH=7 with 0.11 mL of glacial acetic acid, and the reaction solution was concentrated by rotary evaporator, and gel column chromatography (water: methanol = 1:1) was performed to obtain 34.1 mg of white solid 4, with a yield of 81%.

[0034] Furthermore, the alcohol acceptor for the stereoselective glycosylation reaction includes:

[0035]

[0036] Compared with the prior art, the present invention uses 2-deoxy-2-azidogalactosyl PTFAI as a donor, alcohol as an acceptor, and adopts the fusion reagent regulation of TMSI and Ph3PO and the remote participation effect of acyl to synthesize T with high stereoselectivity. N Antigen, avoiding the use of precious metal activators (AgClO4 or Ni(4-F-PhCN)4(OTf)2, etc.) and strong bases (P4-t-Bu, KH, etc.). The experimental operation is simple, the reaction system is mild, no other side reactions occur, the reaction is efficient and stereoselective, and the raw material price is low. DETAILED DESCRIPTION

[0037] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0038] The word "embodiment" used here as an "exemplary" does not necessarily mean that any embodiment described is superior to or better than other embodiments. Unless otherwise specified, the performance index tests in the embodiments of this application are performed using conventional test methods in the art. It should be understood that the terms described in this application are only used to describe specific implementation methods and are not used to limit the content disclosed in this application.

[0039] Unless otherwise specified, the technical and scientific terms used in this document have the same meanings as commonly understood by ordinary technicians in the technical field to which this application belongs; other experimental methods and technical means not specifically specified in this application refer to experimental methods and technical means commonly used by ordinary technicians in this field.

[0040] In order to better illustrate the content of the present application, numerous specific details are provided in the specific examples below. It should be understood by those skilled in the art that the present application can also be implemented without certain specific details. In the embodiments, some methods, means, instruments, equipment, etc. well known to those skilled in the art are not described in detail in order to highlight the main purpose of the present application.

[0041] Under the premise of no conflict, the technical features disclosed in the embodiments of the present application can be combined arbitrarily, and the resulting technical solutions belong to the contents disclosed in the embodiments of the present application.

[0042] Example 1

[0043]

[0044] Under argon protection, 2-deoxy-2-azido-3,4,6-tri-O-benzoyl-α / β-D-pyranose galactose hemiacetal 1 (592.8 mg, 1.15 mmol) and 2,2,2-trifluoro-N-phenylacetyl imide chloride (261.6 mg, 1.26 mmol) were dissolved in 11 mL of acetone, and K2CO3 (238.3 mg, 1.72 mmol) was added. The reaction was allowed to proceed overnight at room temperature, and the reaction was monitored to be complete by thin layer chromatography. The product was filtered through diatomaceous earth, and the filtrate was concentrated by rotary evaporator. The product was subjected to silica gel column chromatography (petroleum ether + 2% triethylamine) to obtain a white solid 2 (774.5 mg) with a yield of 98%.

[0045] Under argon protection, the above white solid 2 (574.9 mg, 0.83 mmol) was dissolved in dry 8.3 mL of dichloromethane, and freshly dried 3A molecular sieves (1 g) and triphenylphosphine oxide (1.39 g, 5.00 mmol) were added in sequence. After stirring at room temperature for 15 min, iodotrimethylsilane (0.12 mL, 0.83 mmol) was added, and the mixture was stirred at room temperature for 1 h. N-benzyloxycarbonyl-L-serine benzyl ester (409.1 mg, 1.25 mmol) was added, and the mixture was moved into a 30°C oil bath for reaction for 5.5 days. Saturated sodium thiosulfate was added to quench the reaction, and the mixture was extracted three times with dichloromethane. The obtained organic phase was washed with saturated sodium chloride in sequence, and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated with a rotary evaporator. Silica gel column chromatography (ethyl acetate / petroleum ether = 1:4) gave a white solid 3 (353.5 mg) with a yield of 51%.

[0046] Data for 3: [α] D 22 = +145.3 (c 0.15, CHCl3). 1 HNMR (400MHz, CDCl3) δ7.91(d,J=7.7Hz,4H),7.79(d,J=7.7Hz,2H),7.53(t,J=7.5Hz,1H ),7.48–7.35(m,5H),7.35–7.19(m,13H),5.89–5.83(m,1H),5.81(d,J=3.2Hz,1H),5.58( dd,J=11.3,3.3Hz,1H),5.20–5.11(m,2H),5.00(d,J=5.2Hz,3H),4.59(d,J=8.1Hz,1H),4 .47–4.39(m,1H),4.32(t,J=6.3Hz,1H),4.19–4.00(m,3H),3.77(dd,J=11.1,3.5Hz,1H). 13 C NMR (151MHz, CDCl3) δ169.6,166.1,165.4,165.3,156.1,136.2,135.1,133.8,133.5,133.3,129.92,129.88,129.5,129.1,129.0,128.81, 128.75,128.7,128.62,128.56,128.52,128.48,128.3,128.2,99.4,69.8,68.6,68.5,68.0,67.7,67.3,62.6,58.2,54.6.HRMS(ESI)calcd forC 45 H 40 N4O 12 [M+Na] +851.2535, found 851.2544.

[0047] Under argon protection, the above white solid 3 was dissolved in 7.6 mL of pyridine, and then thioacetic acid (5.8 mL, 77.84 mmol) was added dropwise, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored to be complete by thin layer chromatography. The reaction solution was concentrated by rotary evaporator, and the intermediate (293.1 mg) was obtained by silica gel column chromatography (ethyl acetate: petroleum ether = 1:2-1:1), with a yield of 89%. The above intermediate (140 mg, 0.17 mmol) was dissolved in 6 mL of methanol, and then 0.6 mL of water, 0.06 mL of glacial acetic acid, and 10% palladium carbon (352.7 mg, 0.33 mmol) were added in sequence. Hydrogen was introduced into the reaction system, and the reaction was allowed to proceed for 6 h at room temperature. The reaction was monitored to be complete by thin layer chromatography. The reaction solution was filtered with a 25 mm diameter polyethersulfone microporous filter membrane, and the filtrate was concentrated by rotary evaporator to obtain 84.8 mg of white solid, with a yield of 83%. Under argon protection, the above white solid was dissolved in 1.5 mL of methanol, and then 1.5 mL of 1,4-dioxane and 2 mL of 1M sodium hydroxide aqueous solution were added in sequence, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored to be complete by thin layer chromatography. The reaction solution was adjusted to pH=7 with 0.11 mL of glacial acetic acid, and the reaction solution was concentrated by rotary evaporator, and gel column chromatography (water: methanol = 1:1) was performed to obtain 34.1 mg of white solid 4, with a yield of 81%.

[0048] Data for 4: [α] D 22 = +178.5 (c 0.16, H2O). 1 HNMR(600MHz,D2O)δ4.92(d,J=3.8Hz,1H),4.20(dd,J=11.1,3.8Hz,1H),4.1 1(dd,J=11.0,2.9Hz,1H),4.01–3.87(m,5H),3.82–3.72(m,2H),2.05(s,3H). 13 C NMR(101MHz,D2O)δ174.3,171.3,97.6,71.0,68.1,67.1,66.3,60.9,54.2,49.3,21.7.HRMS(ESI)calcd for C 11 H 20 N2O8[MH] + 307.1147, found 307.1149.

[0049] In order to further demonstrate the beneficial effects of the present invention and to better understand the present invention, the following experiments are further used to illustrate the T disclosed in the present invention. NThe inventiveness of the method for preparing the antigen should not be construed as a limitation of the present invention; the reaction properties obtained by other experiments conducted by technicians in the field based on the above-mentioned invention content and the applications based on the above-mentioned properties are also considered to fall within the protection scope of the present invention.

[0050] It is worth noting that the following experimental examples of the present invention are all carried out according to route 1.

[0051] Route 1: A mixture of PTFAI donor and acceptor N-(benzyl)benzyloxycarbonyl-5-aminopentanol (1.5 equivalents relative to the donor) was co-evaporated with dry toluene three times. Under argon protection, the mixture was dissolved in freshly distilled dichloromethane (0.1 M) together with triphenylphosphine oxide (6.0 equivalents relative to the donor), and freshly dried The molecular sieve was stirred for 15 minutes. Then iodotrimethylsilane (TMSI) (1.0 equivalent relative to the donor) was added dropwise and reacted at room temperature. After the reaction was completed, the reaction was quenched with a saturated sodium thiosulfate solution, the solution was diluted with dichloromethane, and the organic phase was subsequently washed with water and brine and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The product was purified by column chromatography.

[0052] Experimental Example 1. Optimization of reaction conditions:

[0053]

[0054] Preparation of 13a: 13a was prepared from 11a (129 mg, 0.20 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 6:1) to give 13a (138.4 mg, 89%, α / β = 1.7:1) as yellow syrup.

[0055] data: 1 H NMR (400MHz, CDCl3) δ7.42–7.11(m,25H),5.21–5.11(m,2H),4.91–4.83(m,2H),4.74–4.63(m,2H),4.59–4.37(m,5H),4.02(s,1H ),3.93(s,1H),3.87–3.77(m,2H),3.65–3.51(m,2H),3.50–3.31(m,2H),3.30–3.15(m,2H),1.62–1.46(m,4H),1.36–1.29(m,2H). 13C NMR (101MHz, CDCl3) δ156.7,156.1,138.3,137.95,137.89,137.8,137.7,137.6,136.9,128.52,128. 49,128.46,128.43,128.41,128.3,128.2,128.1,127.9,127.87,127.82,127.78,127.7,127.6,127.2 ,102.4,98.2,80.6,77.3,74.8,74.6,73.5,72.5,72.3,72.2,69.6,68.8,68.5,68.1,67.1,63.4,59. 8,50.5,50.2,47.1,46.2,31.9,29.7,29.4,29.2,29.1,27.9,27.5,23.4,23.2,22.7.HRMS(ESI)calcd for C 47 H 52 N4O7[M+NH4] + 802.4174, found 802.4183.

[0056] Preparation of 13b: 13b was prepared from 11b (118 mg, 0.18 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2.5:1) to give 13b (122.4 mg, 85%, α / β = 2.3:1) as a colorless syrup.

[0057] data: 1 H NMR (400MHz, CDCl3) δ7.45–7.14(m,20H),5.24–5.11(m,2H),4.96–4.82(m ,2H),4.80–4.70(m,2H),4.64–4.54(m,1H),4.49(s,2H),4.23–4.08(m,2H ),3.98–3.79(m,3H),3.68–3.55(m,1H),3.52–3.18(m,4H),2.74–2.64(m, 2H),2.54–2.42(m,2H),2.14(s,3H),1.64–1.47(m,4H),1.39–1.28(m,2H). 13C NMR (101MHz, CDCl3) δ206.4,206.3,172.4,172.3,156.7,156.2,138.1,138.03,137.99,137.6,137.5,136.9,128.59, 128.56,128.54,128.47,128.4,128.33,128.29,128.0,127.95,127.92,127.89,127.84,127.83,127.80,127.3,102. 4,98.2,80.6,77.3,74.7,74.5,73.1,72.7,72.4,72.0,71.8,69.8,68.6,68.2,67.2,63.4,63.3,62.9,59.7,53.5,50 .5,50.3,47.2,46.2,37.9,37.8,29.8,29.7,29.4,29.2,29.1,27.82,27.78,27.5,23.4,23.2,22.7.HRMS(ESI)calcd forC 45 H 52 N4O9[M+NH4] + 810.4073, found 810.4074.

[0058] Preparation of 13c: 13c was prepared from 11c (103 mg, 0.13 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1-7:1) to give 13b (114.5 mg, 95%, α / β = 1.5:1) as yellow syrup.

[0059] data: 1 HNMR(400MHz, CDCl3)δ7.65–7.57(m,4H,Ar),7.44–7.11(m,26H,Ar),5.21–5.1 2(m,2H),4.90(t,J=10.8Hz,1H),4.82(s,1H),4.76–4.67(m,2H),4.58(t,J=10 .4Hz,1H),4.53–4.43(m,2H),3.97–3.87(m,1H),3.83–3.68(m,4H),3.56–3.44 (m,1H),3.38–3.12(m,4H),1.61–1.44(m,4H),1.34–1.26(m,2H),1.05(s,9H). 13C NMR (101MHz, CDCl3) δ138.54,138.46,138.0,137.9,137.8,136.9,135.62,135.59,133.35,133.25,129.9,1 28.63,128.61,128.58,128.52,128.3,128.2,128.1,128.03,128.96,127.92,127.91,127.85,127.81,127.7 ,127.6,127.3,102.5,98.1,80.6,77.4,75.0,74.9,74.8,73.6,72.7,72.6,72.4,71.2,69.7,67.9,67.3,67 .2,63.5,62.7,62.3,60.0,50.6,50.3,47.2,46.3,29.8,29.3,29.2,27.0,23.5,23.3,19.3.HRMS(ESI)calcd for C 56 H 64 N4O7Si[M+NH4] + 950.4883, found 950.4888.

[0060] Preparation of 13d: 13d was prepared from 11d (101.6 mg, 0.15 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1-5:1) to give 13d (111.2 mg, 90%, α / β = 9:1) as a colorless syrup.

[0061] Data: [α] D 25 = +105.1 (c 0.26, CHCl3). 1 H NMR(400MHz, CDCl3)δ8.04(d,J=8.1Hz,2H),7.58–7.15(m,23H),5.92(s,1H),5.23–5.07(m,2H),4.96(s,1H),4.92–4.76(m,1H) ,4.59–4.35(m,5H),4.19(s,1H),4.12–4.00(m,1H),3.79–3.35(m,5H),3.27–3.20(m,2H),1.68–1.47(m,4H),1.42–1.27(m,2H). 13C NMR (101MHz, CDCl3) δ165.7,156.8,156.3,138.0,137.7,137.2,136.9,133.3,130.1,129.9, 129.8,129.6,128.62,128.60,128.5,128.4,128.3,128.00,127.95,127.91,127.89,127.8, 127.4,102.5,98.2,77.4,74.4,73.8,73.7,72.7,71.8,71.7,68.6,68.4,68.2,67.4,67.2,6 6.2,63.1,59.6,50.6,50.3,47.2,46.2,29.8,29.1,27.9,27.5,23.4,23.2.HRMS(ESI)calcd for C 47 H 50 N4O8[M+NH4] + 816.3967, found 816.3968.

[0062] Preparation of 13e: 13e was prepared from 11e (170 mg, 0.26 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1) to give 13e (172 mg, 85%, α / β=4:1) as a colorless syrup.

[0063] data: 1 HNMR(400MHz, CDCl3)δ8.07(d,J=7.7Hz,2H),7.59(t,J=7.4Hz,1H),7.45(t,J=7.7Hz,2H),7.40–7.14 (m,20H),5.55(dd,J=11.1,2.9Hz,1H),5.17(d,J=11.9Hz,2H),4.98(d,J=8.5Hz,1H),4.65(d,J=11.3 Hz,1H),4.53–4.41(m,5H),4.21(s,1H),4.16–4.08(m,1H),3.93(dd,J=11.1,3.5Hz,1H),3.74–3.63( m,1H),3.62–3.53(m,2H),3.48–3.34(m,1H),3.31–3.16(m,2H),1.64–1.48(m,4H),1.40–1.27(m,2H). 13C NMR (101MHz, CDCl3) δ165.8,156.8,156.2,138.0,137.85,137.81,136.9,133.5,130.0,129.4,128.6, 128.5,128.5,128.4,128.3,128.2,128.04,128.96,127.91,127.87,127.83,127.78,127.3,102.4,98. 3,77.4,75.3,75.2,74.9,74.4,73.6,73.5,73.4,71.8,70.0,69.3,68.5,68.2,67.2,61.8,60.4,58.4 ,50.6,50.3,47.2,46.3,41.4,31.6,29.2,28.0,27.7,27.6,23.4,23.2,22.72,22.70.HRMS(ESI)calcd for C 47 H 50 N4O8[M+Na] + 821.3521, found821.3522.

[0064] Preparation of 13f: 13f was prepared from 11f (165.1 mg, 0.24 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give 13f (188.5 mg, 95%, α / β>20:1) as a colorless syrup.

[0065] Data: [α] D 23 = +137.0 (c 0.25, CHCl3). 1 HNMR(400MHz, CDCl3)δ7.98(d,J=7.8Hz,2H),7.88(d,J=7.7Hz,2H),7.58(t,J=7 .4Hz,1H),7.51–7.40(m,3H),7.39–7.12(m,17H),5.93(s,1H),5.78–5.73(m,1H ),5.23–5.07(m,3H),4.50(d,J=10.9Hz,3H),4.42–4.33(m,2H),3.87–3.70(m,2 H),3.64–3.40(m,3H),3.35–3.16(m,2H),1.73–1.48(m,4H),1.45–1.29(m,2H). 13C NMR (101MHz, CDCl3) δ165.4,156.8,156.3,138.0,137.6,136.9,133.4,133.3,129 .9,129.83,129.80,129.5,129.3,128.6,128.5,128.4,128.3,128.2,127.9,127. 8,127.71,127.67,127.3,102.6,98.4,77.4,73.6,69.1,69.0,68.6,68.31,68.27 ,67.2,58.3,50.6,50.4,47.2,46.2,29.7,29.1,27.9,27.6,23.4.HRMS(ESI)calcd for C 47 H 48 N4O9[M+NH4] + 830.3760, found 830.3763.

[0066] Preparation of 13f under reagent-free control conditions: A mixture of donor 11f (132.5 mg, 0.20 mmol) and acceptor N-(benzyl)benzyloxycarbonyl-5-aminopentanol (96.5 mg, 0.29 mmol) was co-evaporated with dry toluene three times. Under argon protection, the above mixture was dissolved in freshly distilled dichloromethane (2 mL) and freshly dried 1% pyrrolidone (20 mL) was added at room temperature. The molecular sieve was stirred for 15 minutes. The reaction was then placed in an ice bath, trimethylsilyl trifluoromethanesulfonate (TMSOTf) was added dropwise, and the reaction was allowed to react at 0°C for 2.5 hours. After the reaction was completed, the reaction was quenched with triethylamine, filtered through diatomaceous earth, and concentrated in vacuo. 13f (148.4 mg, 93%, α / β=2.8:1) was purified by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain 13f as a colorless syrup.

[0067] The above experiments show that when neither C3 nor C4 of the 2-deoxy-2-azidogalactosyl PTFAI donor is protected by acyl groups, the stereoselectivity is poor (α / β=1.5:1~2.3:1)(entry 1-3). When only C3 is protected by Bz, the stereoselectivity is α / β=4:1(entry 5). When only C4 is protected by Bz, the stereoselectivity is α / β=9:1(entry 4). When both C3 and C4 are protected by Bz acyl groups, the best stereoselectivity is achieved (α / β>20:1)(entry 6). When the reaction conditions TMSI and Ph3PO are replaced by TMSOTf, the stereoselectivity is poor (α / β=2.8:1)(entry 7), indicating that reagent regulation (TMSI, Ph3PO) plays a key role in controlling stereoselectivity.

[0068] Experimental Example 2. Stereoselective glycosylation range of 2-deoxy-2-azidogalactosyl PTFAI donor

[0069]

[0070] Preparation of 13g: 13g was prepared from 11g (195.5mg, 0.29mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain 13g (179.0mg, 76%, α / β>20:1) as a colorless syrup.

[0071] Data: [α] D 25 = +174.6 (c 0.17, CHCl3). 1 H NMR(400MHz, CDCl3) δ7.99(d,J=7.8Hz,2H),7.87(d,J=7.7Hz,2H),7.60(t,J=7.4Hz,1H),7.53–7.42(m, 3H),7.42–7.15(m,12H),5.86(s,1H),5.74(d,J=10.8Hz,1H),5.26–5.05(m,3H),4.57–4.48(m,2H),4.4 5–4.33(m,1H),4.30–4.21(m,1H),4.20–4.08(m,1H),3.89–3.70(m,2H),3.60–3.41(m,1H),3.35–3.16( m,2H),2.68(t,J=6.5Hz,2H),2.52(t,J=6.6Hz,2H),2.13(s,3H),1.75–1.50(m,4H),1.47–1.31(m,2H). 13 C NMR (101MHz, CDCl3) δ206.7,206.2,172.1,165.32,165.27,156.6,156.1,149.7,137.9 ,136.8,135.9,133.5,133.3,129.7,129.1,129.0,128.6,128.5,128.41,128.38,128. 3,127.85,127.77,127.2,123.7,98.3,68.7,68.4,67.1,66.9,62.1,58.0,53.5,50.5, 50.3,47.1,46.2,37.7,30.8,29.6,29.0,27.9,27.8,27.7,27.5,23.3.HRMS(ESI)calcd for C 45 H 48 N4O 11[M+NH4] + 838.3658, found 838.3668.

[0072] Preparation of 13h: 13h was prepared from 11h (204 mg, 0.25 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 6:1) to give 13h (215.5 mg, 90%, α / β = 19:1) as a colorless syrup.

[0073] Data: [α] D 25 = +115.1 (c 0.19, CHCl3). 1 H NMR (400MHz, CDCl3) δ7.97(d,J=7.7Hz,2H),7.89(d,J=7.7Hz,2H),7.65(d,J=6.8Hz,2H),7.59 –7.18(m,22H),7.14(t,J=7.4Hz,2H),6.04(d,J=3.4Hz,1H),5.82(d,J=11.0Hz,1H),5.20(d,J =14.8Hz,2H),5.07(d,J=13.1Hz,1H),4.52(d,J=10.7Hz,2H),4.34–4.20(m,1H),3.86–3.60(m ,4H),3.52–3.36(m,1H),3.35–3.18(m,2H),1.70–1.51(m,4H),1.46–1.30(m,2H),1.01(s,9H). 13 C NMR (101MHz, CDCl3) δ165.4,165.2,156.7,156.2,137.9,136.8,135.5,135.4,133.3,13 3.2,132.9,132.7,129.81,129.78,129.73,129.68,129.6,129.3,128.5,128.4,128.2, 127.9,127.81,127.77,127.74,127.6,127.3,127.2,98.3,77.2,69.5,69.1,68.4,67.2 ,61.7,58.3,50.6,50.3,47.1,46.2,29.1,27.9,27.5,26.7,23.4,19.0.HRMS(ESI)calcd forC 56 H 60 N4O9Si[M+NH4] + 978.4468, found 978.4469.

[0074] Preparation of 13i: 13i was prepared from 11i (202.0 mg, 0.32 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give 13i (193.4 mg, 78%, α / β>20:1) as a colorless syrup.

[0075] Data: [α] D 25 = +176.1 (c 0.26, CHCl3). 1 H NMR (400MHz, CDCl3) δ8.00(d,J=7.7Hz,2H),7.87(d,J=7.7Hz,2H),7.58(t,J=7.4Hz,1H),7.51– 7.41(m,3H),7.40–7.17(m,12H),5.89(s,1H),5.75(d,J=10.9Hz,1H),5.24–5.07(m,3H),4.52(d ,J=6.7Hz,2H),4.43–4.34(m,1H),4.26–4.12(m,2H),3.87(dd,J=11.1,3.4Hz,1H),3.81–3.68( m,1H),3.59–3.43(m,1H),3.35–3.19(m,2H),1.99(s,3H),1.75–1.50(m,4H),1.46–1.31(m,2H). 13 CNMR(101MHz, CDCl3)δ170.3,165.4,156.7,156.2,137.9,136.8,133.6,133.3,129.9,129.8,129.2,129.1,128.6,128.53,128.45,128.4,128 .3,127.9,127.8,127.3,98.4,68.8,68.7,68.5,67.2,67.0,62.1,58.1,50.6,50.3,47.1,46.2,29.1,27.9,27.5,23.4,20.6.HRMS(ESI)calcd for C 42 H 44 N4O 10 [M+H] + 765.3130, found 765.3135.

[0076] Preparation of 13j: 13j was prepared from 11j (222.5 mg, 0.32 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=4:1) to give 13j (223.1 mg, 83%, α / β=19:1) as a colorless syrup.

[0077] Data: [α] D 25 = +136.3 (c 0.28, CHCl3). 1 HNMR(400MHz, CDCl3)δ8.01(m,4H),7.88(d,J=7.7Hz,2H),7.60–7.53(m,1H),7.51– 7.40(m,4H),7.40–7.15(m,14H),6.01(s,1H),5.82(d,J=10.4Hz,1H),5.25–5.11(m ,3H),4.62–4.45(m,4H),4.36(d,J=10.6Hz,1H),3.92(d,J=10.8Hz,1H),3.82–3.68 (m,1H),3.58–3.42(m,1H),3.33–3.17(m,2H),1.74–1.45(m,4H),1.41–1.27(m,2H). 13 C NMR (101MHz, CDCl3) δ165.9,165.33,165.27,156.6,156.1,137.9,136.8,133.5 ,133.3,133.2,129.9,129.8,129.73,129.65,129.6,129.4,129.1,129.0,128. 6,128.5,128.4,128.3,127.8,127.2,98.3,68.8,68.7,68.6,67.1,67.1,62.5, 58.1,50.5,50.2,47.0,46.1,29.6,29.0,27.8,27.4,23.3.HRMS(ESI)calcdfor C 47 H 46 N4O 10 [M+NH4] + 844.3552, found 844.3560.

[0078] Preparation of 13k: 13k was prepared from 11k (143.8 mg, 0.26 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give 13k (143.9 mg, 82%, α / β = 16:1) as a colorless syrup.

[0079] Data: [α] D 25 = +98.9 (c 0.17, CHCl3). 1H NMR (400MHz, CDCl3) δ7.32–6.99(m,15H),5.42(d,J=3.3Hz,1H),5.28(dd,J=11.1 ,3.3Hz,1H),5.09(d,J=10.5Hz,2H),4.86(d,J=10.4Hz,1H),4.48–4.28(m,4H),4. 11–4.01(m,1H),3.66–3.54(m,1H),3.49(dd,J=11.1,3.5Hz,1H),3.43–3.25(m,3H ),3.22–3.08(m,2H),1.95(d,J=2.6Hz,6H),1.58–1.36(m,4H),1.32–1.20(m,2H). 13 C NMR (101MHz, CDCl3) δ170.0,169.8,156.7,156.2,138.0,137.6,136.9,128.6,128.5, 128.4,127.92,127.88,127.82,127.80,127.3,102.4,98.1,73.6,73.5,72.1,71.2,7 0.3,68.5,68.4,68.2,67.9,67.8,67.5,67.2,67.0,61.2,57.6,50.6,50.3,47.1,46. 2,29.4,29.2,29.0,27.9,27.5,23.4,23.2,22.7,20.70,20.66,20.6.HRMS(ESI)calcd for C 37 H 44 N4O9[M+NH4] + 706.3447, found 706.3449.

[0080] Preparation of 131: 131 was prepared from 111 (132.7 mg, 0.19 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give 131 (120.8 mg, 76%, α / β = 12:1) as a colorless syrup.

[0081] Data: [α] D 25 = +62.5 (c 0.25, CHCl3). 1H NMR(400MHz, CDCl3) δ7.62(t,J=7.1Hz,4H),7.43–7.15(m,16H),5.59(d,J=3.3Hz,1H),5. 40(dd,J=11.2,3.2Hz,1H),5.18(d,J=12.0Hz,2H),4.90(d,J=11.6Hz,1H),4.50(d,J=8.5H z,2H),4.10–4.02(m,1H),3.73–3.59(m,3H),3.55(dd,J=11.2,3.5Hz,1H),3.42–3.16(m,3 H),2.04(s,3H),2.00(d,J=3.8Hz,3H),1.65–1.47(m,4H),1.39–1.28(m,2H),1.03(s,9H). 13 C NMR (101MHz, CDCl3) δ169.84,169.80,156.7,156.2,137.9,136.8,135.6,135.5,133 .0,132.9,129.9,129.8,128.52,128.50,128.4,127.84,127.81,127.8,127.7,127. 3,102.3,98.0,73.2,69.1,68.4,68.3,67.7,67.1,66.4,61.6,61.2,57.6,53.4,50. 6,50.3,47.1,46.2,29.0,27.9,27.5,26.7,23.3,20.7,20.6,19.0.HRMS(ESI)calcd for C 46 H 56 N4O9Si[M+NH4] + 854.4155, found854.4156.

[0082] Preparation of 13m: 13m was prepared from 11m (201 mg, 0.40 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give 13m (167 mg, 68%, α / β = 11:1) as a colorless syrup.

[0083] Data: [α] D 25 = +95.1 (c 0.46, CHCl3). 1H NMR (400MHz, CDCl3) δ7.33–7.04(m,10H),5.36(s,1H),5.27(d,J=11.5Hz,1H),5.08(d,J=12.9Hz,2H),4.86(s,1H),4.47–4.35(m,2H),4.15–3.9 3(m,3H),3.64–3.46(m,2H),3.43–3.26(m,1H),3.25–3.10(m,2H),2.04 (s,3H),1.95(s,3H),1.92(s,3H),1.59–1.38(m,4H),1.32–1.18(m,2H). 13 C NMR (101MHz, CDCl3) δ170.1,169.92,169.89,169.6,156.6,156.1,137.9,136 .8,128.4,128.3,127.84,127.81,127.7,127.2,102.2,98.0,70.9,70.5,70.2 ,68.5,68.1,67.6,67.0,66.5,66.3,61.6,61.2,60.9,57.3,50.5,50.3,47.0, 46.1,29.6,29.0,28.9,27.8,27.4,23.2,23.0,20.51,20.46.HRMS(ESI)calcd for C 32 H 40 N4O 10 [M+H] + 641.2817, found 641.2810.

[0084] Preparation of 13n: 13n was prepared from 11n (300 mg, 0.45 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give 13n (225 mg, 85%, α / β>20:1) as a colorless syrup.

[0085] Data: [α] D 24 = +107.3 (c 0.24, CHCl3). 1H NMR(400MHz, CDCl3) δ7.95(d,J=7.7Hz,2H),7.54(t,J=7.4Hz,1H),7.39(t,J=7.6Hz,2H),7.33–7.06(m,15H),5.67 (d,J=3.2Hz,1H),5.39(dd,J=11.2,3.2Hz,1H),5.11(d,J=12.0Hz,2H),4.95(d,J=11.4Hz,1H),4.46–4.38(m,3H),4 .29(d,J=11.9Hz,1H),4.24–4.15(m,1H),3.69–3.57(m,2H,H-2),3.49–3.30(m,3H),3.25–3.06(m,2H),2.77–2.66( m,1H),2.65–2.57(m,1H),2.56–2.45(m,1H),2.44–2.34(m,1H),2.05(s,3H),1.61–1.41(m,4H),1.32–1.21(m,2H). 13 C NMR (101MHz, CDCl3) δ206.0,171.7,165.6,156.7,156.2,138.0,137.5,136.9,1 33.5,129.9,129.8,129.4,128.7,128.62,128.56,128.5,128.3,127.94,127.88 ,127.8,127.7,127.6,127.3,98.3,77.4,73.5,68.84,68.76,68.6,68.1,67.2,5 7.9,53.5,50.6,50.4,47.1,46.2,37.9,29.7,29.1,28.0,23.4.HRMS(ESI)calcd for C 45 H 50 N4O 10 [M+NH4] + 824.3865, found824.3872.

[0086] Preparation of 13o: 13o was prepared from 11o (306 mg, 0.5 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) and then by gel column LH-20 (dichloromethane: methanol = 1:1) to obtain 13o (309 mg, 84%, α / β = 20:1) as a colorless syrup.

[0087] Data: [α] D 24 = +92.9 (c 0.61, CHCl3).1 H NMR (400MHz, CDCl3) δ8.04(d,J=7.8Hz,2H),7.59(t,J=7.5Hz,1H),7.45(t,J=7.6Hz,2H),7 .40–7.13(m,15H),5.80–5.73(m,1H),5.52–5.43(m,1H),5.18(d,J=11.8Hz,2H),5.03(d,J= 10.9Hz,1H),4.54–4.44(m,3H),4.36(d,J=12.0Hz,1H),4.32–4.23(m,1H),3.77–3.65(m,2H ),3.59–3.39(m,3H),3.33–3.17(m,2H),2.00(s,3H),1.71–1.45(m,4H),1.41–1.27(m,2H). 13 C NMR (101MHz, CDCl3) δ170.0,165.6,156.8,156.3,138.0,137.6,133.5,129.9,1 29.6,128.64,128.62,128.54,128.51,128.43,128.40,128.0,127.95,127.93, 127.87,127.82,127.75,127.69,127.4,98.3,73.6,68.8,68.7,68.20,68.15,6 7.3,57.9,50.7,50.4,47.2,46.3,29.1,28.0,27.6,23.4,20.8.HRMS(ESI)calcd for C 42 H 46 N4O9[M+H] + 751.3338, found 751.3334.

[0088] Preparation of 13p: 13p was prepared from 11p (240 mg, 0.36 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1) to give 13p (250 mg, 86%, α / β = 17: 1) as a colorless syrup.

[0089] Data: [α] D 24 = +74.8 (c 0.17, CHCl3). 1H NMR (400MHz, CDCl3) δ7.48–6.96(m,15H),5.50(d,J=3.3Hz,1H),5.33(dd,J=11.0,3.2Hz, 1H),5.17(d,J=10.5Hz,2H),4.94(d,J=10.4Hz,1H),4.54–4.40(m,4H),4.19–4.07(m,1H) ,3.73–3.55(m,2H),3.53–3.33(m,3H),3.30–3.15(m,2H),2.84–2.67(m,4H),2.66–2.56( m,2H),2.56–2.45(m,2H),2.17(s,3H),2.16(s,3H),1.66–1.46(m,4H),1.39–1.21(m,2H). 13 C NMR (101MHz, CDCl3) δ206.2,206.1,171.9,171.8,156.7,156.2,138.0,137. 8,136.9,128.6,128.5,128.44,128.40,128.0,127.9,127.84,127.77,127. 3,98.1,73.5,68.7,68.5,68.3,68.0,67.8,67.2,57.7,50.6,50.3,47.1,46 .2,37.9,37.9,37.8,29.85,29.77,29.1,27.9,27.7,23.4.HRMS(ESI)calcd for C 43 H 52 N4O 11 [M+NH4] + 818.3971, found 818.3979.

[0090] The above experiments show that when the C3 and C4 positions of the 2-deoxy-2-azidogalactosyl PTFAI donor are both acyl protected (Ac, Bz, Lev), it often has better stereoselectivity (α / β=11:1~>20:1).

[0091] Experimental Example 3. Alcohol acceptor range for stereoselective glycosylation

[0092]

[0093] Preparation of 15a: 15a was prepared from 11f (153.8 mg, 0.23 mmol) and acceptor 14a (92.5 mg, 0.34 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to give 15a (152.6 mg, 92%, α / β>20:1) as a colorless syrup.

[0094] Data: [α] D 23 = +149.3 (c 0.16, CHCl3). 1 H NMR (400MHz, CDCl3) δ7.98(d,J=7.6Hz,2H),7.88(d,J=7.6Hz,2H),7.57(t,J=7.4Hz,1H),7.51–7.39 (m,3H),7.30(t,J=7.7Hz,2H),7.25–7.13(m,5H),5.94(d,J=3.3Hz,1H),5.77(dd,J=11.1,3.3Hz,1H) ,5.14(d,J=3.4Hz,1H,H-1),4.52(d,J=11.9Hz,1H,CH2-Bn),4.44–4.37(m,2H),3.87–3.77(m,2H),3. 64–3.51(m,3H),1.74–1.65(m,2H),1.41(q,J=7.3Hz,2H),1.30–1.25(m,28H),0.88(t,J=6.7Hz,3H). 13 C NMR (101MHz, CDCl3) δ165.4,137.7,133.4,133.2,129.9,129.8,129.6,129.4,128.6,128.42,128.37,128.3,127.71,127.67,98.5,73. 6,69.2,69.1,69.0,68.32,68.28,58.4,32.0,29.80,29.78,29.75,29.73,29.67,29.53,29.50,29.4,26.3,22.8,14.2.HRMS(ESI)calcd for C 45 H 61 N3O7[M+NH4] + 773.4848, found 773.4857.

[0095] Preparation of 15b: 15b was prepared from 11f (157.7 mg, 0.23 mmol) and acceptor 14b (35.0 mg, 0.35 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to give 15b (117 mg, 86%, α / β>20:1) as a colorless syrup.

[0096] Data: [α] D 21 = +204.5 (c 0.21, CHCl3). 1 H NMR (400MHz, CDCl3) δ7.88(d,J=7.8Hz,2H),7.79(d,J=7.7Hz,2H),7.51(t,J=7.4Hz ,1H),7.44–7.32(m,3H),7.23(t,J=7.7Hz,2H),7.17–7.07(m,5H),5.87(d,J=3.2Hz, 1H),5.66(dd,J=11.1,3.3Hz,1H),5.16(d,J=3.5Hz,1H),4.42(d,J=11.9Hz,1H),4. 35–4.28(m,2H),4.06–3.91(m,2H),3.87(dd,J=11.1,3.6Hz,1H),3.56–3.46(m,2H). 13 C NMR (101MHz, CDCl3) δ165.4,165.3,137.5,133.6,133.4,130.0,129.90,129.85,129.4,129.2,128.7,128.5,128 .4,127.8,127.7,124.9,122.2,99.1,73.6,69.2,68.8,68.6,68.0,65.7,65.3,65.0,64.6,58.0.HRMS(ESI)calcd for C 29 H 26 N3O7F3[M+NH4] + 603.2061, found 603.2071.

[0097] Preparation of 15c: 15c was prepared from 11f (163.2 mg, 0.24 mmol) and acceptor 14c (39.2 mg, 0.36 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1-5:1) to give 15c (135.6 mg, 94%, α / β>20:1) as a colorless syrup.

[0098] Data: [α] D 23= +211.3 (c 0.14, CHCl3). 1 H NMR (400MHz, CDCl3) δ7.98(d,J=7.7Hz,2H),7.89(d,J=7.7Hz,2H),7.58(t,J=7.4Hz,1H),7 .50–7.40(m,5H),7.40–7.28(m,6H),7.26–7.16(m,4H),5.94(d,J=3.3Hz,1H),5.81(dd,J=1 1.0,3.3Hz,1H),5.24(d,J=3.5Hz,1H),4.84(d,J=11.9Hz,1H),4.69(d,J=11.9Hz,1H),4.5 2(d,J=12.0Hz,1H),4.47–4.35(m,2H),3.92(dd,J=11.0,3.5Hz,1H),3.59(d,J=6.4Hz,2H). 13 C NMR (101MHz, CDCl3) δ165.4,137.7,136.5,133.4,133.3,129.85,129.82,129.5,129.3,128.64,128.60,128.5, 128.4,128.35,128.26,128.20,127.72,127.68,97.2,73.6,70.0,69.3,69.0,68.5,68.3,58.4.HRMS(ESI)calcd for C 34 H 31 N3O7[M+NH4] + 611.2500, found 611.2501.

[0099] Preparation of 15d: 15d was prepared from 11f (155.4 mg, 0.23 mmol) and acceptor 14d (42.2 mg, 0.35 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1-5:1) to give 15d (129.1 mg, 92%, α / β>20:1) as a colorless syrup.

[0100] Data: [α] D 23 = +222.4 (c 0.27, CHCl3). 1H NMR (400MHz, CDCl3) δ7.98(d,J=7.7Hz,2H),7.90(d,J=7.7Hz,2H),7.59(t,J=7.4Hz,1H) ,7.50(t,J=7.4Hz,1H),7.44(t,J=7.7Hz,2H),7.37–7.15(m,12H),5.83(d,J=3.2Hz,1H) ,5.74(dd,J=11.0,3.3Hz,1H),5.18(d,J=3.4Hz,1H),4.47(d,J=12.0Hz,1H),4.35(d,J= 11.9Hz,1H),4.07–3.95(m,2H),3.90–3.78(m,2H),3.55–3.45(m,2H),3.08–2.98(m,2H). 13 C NMR (101MHz, CDCl3) δ165.40,165.38,138.4,137.7,133.4,133.3,129.84,129.81,129.5,129.3,129.1,128.5 9,128.57,128.4,127.7,127.62,126.60,98.1,73.4,69.2,69.1,69.0,68.3,68.2,58.4,36.1.HRMS(ESI)calcd for C 35 H 33 N3O7[M+NH4] + 625.2657, found 611.2660.

[0101] Preparation of 15e: 15e was prepared from 11f (156.5 mg, 0.23 mmol) and acceptor 14e (30.0 mg, 0.35 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1-10:1) to give 15e (121.4 mg, 91%, α / β>20:1) as a colorless syrup.

[0102] Data: [α] D 21 = +227.4 (c 0.13, CHCl3). 1H NMR (400MHz, CDCl3) δ7.98(d,J=7.7Hz,2H),7.88(d,J=7.7Hz,2H),7.60(t,J=7.4Hz,1H),7.47(m,7.5H z,3H),7.32(t,J=7.7Hz,2H),7.22(t,J=7.4Hz,5H),5.93(d,J=3.3Hz,1H),5.89–5.80(m,1H),5.77(dd, J=11.1,3.3Hz,1H),5.15(d,J=3.4Hz,1H),5.08(d,J=17.2Hz,1H),5.01(d,J=10.2Hz,1H),4.53(d,J=11 .9Hz,1H),4.44–4.37(m,2H),3.84(m,2H),3.65–3.53(m,3H),2.21(q,J=7.2Hz,2H),1.84–1.75(m,2H). 13 C NMR (101MHz, CDCl3) δ165.5,165.4,137.9,137.7,133.5,133.3,129.90,129.87,129.6,129.4,128.6,128.42 ,128.39,127.8,127.7,115.3,98.5,73.6,69.13,69.05,68.4,68.3,68.2,58.4,30.4,28.7.HRMS(ESI)calcd for C 32 H 33 N3O7[M+NH4] + 589.2657, found 589.2663.

[0103] Preparation of 15f: 15f was prepared from 11f (152 mg, 0.23 mmol) and acceptor 14f (44.7 mg, 0.34 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1-5: 1) to give 15f (129.8 mg, 93%, α / β> 20: 1) as a white solid.

[0104] Data: [α] D 23 = +195.6 (c 0.18, CHCl3). 1H NMR(400MHz, CDCl3)δ8.01–7.95(m,2H),7.91–7.84(m,2H),7.59(t,J=7.4Hz,1H),7.53–7.41(m,3H),7 .32(t,J=7.7Hz,2H),7.26–7.14(m,5H),5.94(d,J=3.2Hz,1H),5.75(dd,J=11.0,3.4Hz,1H),5.24(d,J= 3.5Hz,1H),4.52(d,J=11.9Hz,1H),4.48–4.34(m,3H),4.12(dd,J=8.4,6.3Hz,1H),3.90(dd,J=11.0,3. 5Hz,1H),3.86–3.79(m,2H),3.72(dd,J=10.6,5.3Hz,1H),3.65–3.54(m,2H),1.47(s,3H),1.38(s,3H). 13 C NMR (101MHz, CDCl3) δ165.40,165.38,137.6,133.5,133.3,129.9,129.8,129.5,129.3,128.6,128.39,128.37,1 27.73,127.71,109.7,98.7,74.4,73.6,69.14,69.05,68.9,68.5,68.2,66.8,58.4,26.9,25.5.HRMS(ESI)calcd forC 33 H 35 N3O9[M+NH4] + 635.2712, found 635.2716.

[0105] Preparation of 15g: 15g was prepared from 11f (193.4 mg, 0.29 mmol) and acceptor 14g (88.8 mg, 0.19 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give 15g (184 mg, 99%, α / β>20:1) as a white solid.

[0106] Data: [α] D 23 = +149.1 (c 0.15, CHCl3). 1H NMR (400MHz, CDCl3) δ7.94(d,J=7.7Hz,2H),7.86(d,J=7.7Hz,2H),7.53(t,J=7.5Hz,1H),7.46 –7.22(m,20H),7.22–7.13(m,5H),5.87(s,1H),5.68(d,J=11.0Hz,1H),5.22(s,1H),4.97(dd, J=15.9,11.2Hz,2H),4.85–4.72(m,2H),4.69–4.57(m,3H),4.43(d,J=12.0Hz,1H,CH2-Bn),4. 32(d,J=11.7Hz,2H),4.02(t,J=9.4Hz,1H),3.91–3.73(m,4H),3.62–3.46(m,4H),3.39(s,3H). 13 C NMR (101MHz, CDCl3) δ165.3,138.8,138.3,138.2,137.6,133.4,133.2,1 29.8,129.7,129.5,129.3,128.53,128.46,128.4,128.35,128.29,128.1 ,127.91,127.87,127.8,127.6,127.5,98.5,98.1,82.1,80.1,77.8,75.7 ,75.1,73.42,73.37,70.1,68.9,68.1,66.7,58.5,55.2.HRMS(ESI)calcd for C 55 H 55 N3O 12 [M+NH4] + 967.4124, found 967.4134.

[0107] Preparation of 15h: 15h was prepared from 11f (162.7 mg, 0.24 mmol) and acceptor 14h (89.5 mg, 0.16 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give 15h (151.1 mg, 90%, α / β>20:1) as a white solid.

[0108] Data: [α] D 23 = +122.8 (c 0.24, CHCl3). 1H NMR(400MHz, CDCl3)δ8.02–7.96(m,2H),7.92–7.87(m,2H),7.59(t,J=7.5Hz,1H),7.53–7.38(m,8H) ,7.37–7.25(m,15H),7.24–7.14(m,6H),5.78(d,J=3.3Hz,1H),5.65(dd,J=11.0,3.3Hz,1H),5.27(d, J=3.5Hz,1H),4.96–4.84(m,4H),4.76–4.66(m,3H),4.49–4.37(m,2H),4.28(t,J=6.3Hz,1H),3.88( dd,J=11.1,3.4Hz,3H),3.74(t,J=8.8Hz,1H),3.64(t,J=9.3Hz,1H),3.60–3.45(m,4H),2.29(s,3H). 13 CNMR (101MHz, CDCl3) δ165.5,165.3,138.5,138.23,138.19,137.85,137.83,133.5,133.3,13 2.1,130.0,129.91,129.88,129.84,129.7,129.4,128.65,128.63,128.53,128.50,128.43,12 8.38,128.3,128.02,127.98,127.92,127.89,127.78,127.69,127.66,98.6,87.5,87.0,81.0 ,78.8,77.7,75.9,75.5,75.2,73.4,69.2,69.0,68.3,68.1,66.8,58.7,21.2.HRMS(ESI)calcd for C 61 H 59 N3O 11 S[M+NH4] + 1059.4209, found 1059.4205.

[0109] Preparation of 15i: 15i was prepared from 11f (147.4 mg, 0.22 mmol) and acceptor 14i (87.2 mg, 0.15 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give 15i (135.6 mg, 86%, α / β>20:1) as a white solid.

[0110] Data: [α] D 25 = +182.7 (c 0.20, CHCl3).1 H NMR(400MHz,CDCl3)δ7.98(dd,J=11.0,7.8Hz,4H),7.90(dd,J=10.9,7.9Hz,4H),7.76(d,J=7.8Hz,2H),7.58(t,J=7.5Hz,2H),7.50(t,J=8.0Hz,4H),7.46–7.36(m,7H),7.32(t,J=7.7Hz,2H),7.26–7.15(m,9H),5.98(d,J=3.2Hz,1H),5.85(d,J=3.2Hz,1H),5.79(t,J=9.9Hz,1H),5.71(dd,J=11.0,3.2Hz,1H),5.62(dd,J=10.0,3.2Hz,1H),5.12–5.06(m,2H),4.49(dd,J=12.2,5.7Hz,2H),4.41(d,J=12.1Hz,1H),4.36–4.31(m,1H),4.07(dd,J=10.5,7.0Hz,1H),3.89(dd,J=11.0,3.6Hz,1H),3.79(dd,J=10.5,4.4Hz,1H),3.64–3.51(m,2H),2.37(s,3H). 13 C NMR(101MHz,CDCl3)δ165.6,165.5,165.42,165.35,165.26,138.6,137.8,133.6,133.48,133.46,133.39,133.31,133.26,130.1,130.0,129.94,129.90,129.86,129.6,129.5,129.4,129.1,129.0,128.6,128.5,128.4,128.3,127.9,127.72,127.68,98.7,85.9,76.6,73.5,73.2,69.2,69.0,68.5,68.3,68.2,68.0,58.5,21.3.HRMS(ESI)calcd forC 61 H 53 N3O 14 S[M+NH4] + 1101.3586,found 1101.3589.

[0111] Preparation of 15j: 15j was prepared from 11f (227.8 mg, 0.33 mmol) and acceptor 14j (57.2 mg, 0.22 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give 15j (162 mg, 99%, α / β>20:1) as a yellow solid.

[0112] Data: [α] D 22 = +115.1 (c 0.15, CHCl3). 1 H NMR (400MHz, CDCl3) δ7.98(d,J=7.7Hz,2H),7.88(d,J=7.7Hz,2H),7.58(t,J=7.4Hz,1H),7.52–7.39(m,3H),7.31 (t,J=7.7Hz,2H),7.25–7.13(m,5H),5.95(d,J=3.3Hz,1H),5.78(dd,J=11.1,3.3Hz,1H),5.54(d,J=5.0Hz,1H),5 .22(d,J=3.4Hz,1H),4.64(dd,J=7.8,2.4Hz,1H),4.55–4.48(m,2H),4.43–4.31(m,3H),4.10(t,J=6.5Hz,1H),3. 95(dd,J=10.1,6.3Hz,1H),3.89–3.80(m,2H),3.60(m,J=9.7,6.3Hz,2H),1.59(s,3H),1.45(s,3H),1.34(s,6H). 13 C NMR (101MHz, CDCl3) δ165.4,137.7,133.4,133.2,129.9,129.8,129.6,129.4,128.6,128.4,128.3,127.7,127.6,109.4 ,108.8,98.7,96.4,73.4,71.0,70.8,69.0,68.9,68.1,68.0,67.4,66.5,58.5,26.2,26.1,25.1,24.5.HRMS(ESI)calcd for C 39 H 43 N3O 12 [M+NH4] + 763.3185, found 763.3192.

[0113] Preparation of 15k: 15k was prepared from 11f (153.3 mg, 0.23 mmol) and acceptor 14k (70.3 mg, 0.15 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1-3:1) to give 15k (132.3 mg, 92%, α / β>20:1) as a yellow solid.

[0114] Data: [α] D 23 = +166.9 (c 0.17, CHCl3). 1 H NMR (400MHz, CDCl3) δ7.97(d,J=7.7Hz,2H),7.87(d,J=7.7Hz,2H),7.58(t,J=7.4Hz,1H),7.51–7.41(m,3H),7.3 9–7.25(m,18H),7.23–7.18(m,3H),7.17–7.12(m,1H),5.93(d,J=3.2Hz,1H),5.76(dd,J=11.0,3.3Hz,1H),5.29 (d,J=3.4Hz,1H),4.99(d,J=11.2Hz,1H),4.77–4.67(m,3H),4.67–4.60(m,3H),4.52–4.46(m,2H),4.38(d,J=11 .9Hz,1H,CH2-Bn),3.96–3.89(m,4H),3.88–3.83(m,2H),3.81(d,J=2.0Hz,1H),3.64–3.51(m,2H),3.37(s,3H). 13 C NMR (101MHz, CDCl3) δ165.4,138.7,138.6,138.5,137.7,133.4,133.3,129.90, 129.88,129.7,129.5,128.6,128.49,128.46,128.44,128.40,128.38,128.0,1 27.90,127.85,127.80,127.73,127.70,127.65,99.1,98.2,80.5,75.2,74.9,7 3.5,72.9,72.2,71.5,69.1,69.0,68.1,68.0,67.2,58.5,54.9.HRMS(ESI)calcd for C 55 H 55 N3O 12 [M+NH4] + 967.4124, found 967.4130.

[0115] Preparation of 15l: 15l was prepared from 11f (159.9 mg, 0.24 mmol) and acceptor 14l (94.2 mg, 0.16 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1-3.5:1) to give 15l (158.9 mg, 93%, α / β>20:1) as a white solid.

[0116] Data: [α] D 23 = +42.8 (c 0.13, CHCl3). 1 H NMR (400MHz, CDCl3) δ8.17–8.11(m,2H),8.05–8.00(m,2H),7.97–7.92(m,2H),7.92–7.84(m,4H),7.60–7.40(m,10H),7.35(q,J=7.7Hz,4H), 7.31–7.24(m,2H),7.21–7.15(m,2H),7.14–7.06(m,4H),6.98–6.93(m ,2H),6.13(dd,J=10.1,3.3Hz,1H),6.07(t,J=10.0Hz,1H),5.91–5.88( m,2H),5.76(dd,J=11.0,3.3Hz,1H),5.69(d,J=1.8Hz,1H),5.16(d,J=3.5Hz,1H),4.62–4.55(m,1H),4.37(t,J=6.5Hz,1H),4.33(d,J=12.1H z,1H),4.20(d,J=12.1Hz,1H),4.09(dd,J=11.0,6.0Hz,1H),3.91(dd,J=11.0,3.4Hz,1H),3.80–3.79(m,1H),3.78(s,3H),3.49–3.37(m,2H). 13C NMR (101MHz, CDCl3) δ165.8,165.71,165.67,165.4,165.2,155.9,150.1,137.6,133.62 ,133.60,133.4,133.3,130.1,130.0,129.92,129.89,129.87,129.6,129.5,129.3,129. 2,129.1,128.7,128.63,128.61,128.4,128.3,127.63,127.60,118.5,115.0,98.3,97.4 ,73.4,70.7,70.23,70.18,69.5,69.0,68.2,68.1,67.2,58.6,55.8.HRMS(ESI)calcdfor C 61 H 53 N3O 16 [M+NH4] + 1101.3764, found 1101.3766.

[0117] Preparation of 15m: 15m was prepared from 11f (164.1 mg, 0.24 mmol) and acceptor 14m (73.1 mg, 0.16 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give 15m (149.2 mg, 98%, α / β>20:1) as a white solid.

[0118] Data: [α] D 22 = +199.5 (c 0.20, CHCl3). 1H NMR (400MHz, CDCl3) δ8.05(d,J=7.7Hz,2H),7.96(d,J=7.7Hz,2H),7.89(d,J=7.8Hz,2H),7.59(t,J=7.4Hz,1H),7.54– 7.36(m,11H),7.33(m,4H),7.29–7.22(m,1H),7.20–7.09(m,6H),5.81(d,J=3.3Hz,1H),5.63(dd,J=11.1,3.3Hz,1H),5 .59(d,J=3.0Hz,2H),5.21(d,J=3.5Hz,1H),4.83(d,J=11.8Hz,1H),4.65–4.57(m,2H),4.44(d,J=12.0Hz,1H),4.34(d ,J=12.0Hz,1H),4.27(dd,J=6.6,2.3Hz,1H),4.20(t,J=6.3Hz,1H),3.94–3.83(m,3H),3.56–3.44(m,2H),2.32(s,3H). 13 C NMR (101MHz, CDCl3) δ165.6,165.4,165.2,138.1,137.7,137.3,133.53,133.4 9,133.3,133.2,130.1,130.0,129.90,129.86,129.6,129.4,128.7,128.65,12 8.60,128.5,128.43,128.41,128.2,127.73,127.71,98.5,91.4,82.64,82.58 ,80.8,73.5,72.8,69.2,69.0,68.33,68.27,66.5,58.6,21.2.HRMS(ESI)calcd for C 53 H 49 N3O 11 S[M+NH4] + 953.3426, found 953.3418.

[0119] Preparation of 15n: 15n was prepared from 11f (127.3 mg, 0.19 mmol) and acceptor 14n (93.2 mg, 0.28 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give 15n (112.3 mg, 73%, α / β>20:1)) as a yellow solid.

[0120] Data: [α] D 25= +168.8 (c 0.17, CHCl3). 1 H NMR (400MHz, CDCl3) δ7.94(d,J=7.8Hz,2H),7.87(d,J=7.7Hz,2H),7.59(t,J=7.4Hz,1H),7.50(t,J=7.5Hz,1H), 7.44(t,J=7.7Hz,2H),7.40–7.28(m,12H),7.22–7.14(m,5H),6.10(d,J=8.5Hz,1H),5.81(d,J=3.3Hz,1H),5.60 (dd,J=11.1,3.3Hz,1H),5.24(s,2H),5.16–5.07(m,2H),5.05(d,J=3.6Hz,1H),4.67(d,J=8.2Hz,1H),4.47(d,J =12.2Hz,1H),4.36–4.23(m,3H),4.06(dd,J=10.9,3.0Hz,1H),3.80(dd,J=11.1,3.6Hz,1H),3.57–3.44(m,2H). 13 C NMR (101MHz, CDCl3) δ169.6,165.4,165.3,156.2,137.5,136.3,135.2,133.5,133.4,129.9,129.8,129.4,129.2,128.8,128.69,128. 65,128.60,128.43,128.40,128.36,128.3,127.7,99.9,73.5,70.5,68.8,68.75,68.71,68.0,67.9,67.3,58.3,54.8.HRMS(ESI)calcd forC 45 H 42 N4O 11 [M+NH4] + 832.3188, found 832.3192.

[0121] Preparation of 15o: 15o was prepared from 11f (148.6 mg, 0.22 mmol) and acceptor 14o (68.2 mg, 0.15 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1-3:1) to give 15o (126.3 mg, 91%, α / β>20:1)) as a yellow solid.

[0122] Data: [α] D 23 = +156.6 (c 0.14, CHCl3). 1H NMR (400MHz, CDCl3) δ7.92(d,J=7.7Hz,2H),7.88(d,J=7.7Hz,2H),7.56(t,J=7.5Hz,1H),7.49(t,J=7.4Hz,1H),7.44–7. 25(m,20H),7.14–7.06(m,4H),5.94(d,J=3.8Hz,1H),5.87(d,J=3.2Hz,1H),5.66(dd,J=11.2,3.2Hz,1H),5.15(d,J=11.0 Hz,1H),4.89(d,J=11.0Hz,1H),4.73(d,J=12.0Hz,1H),4.67–4.60(m,3H),4.54(d,J=12.3Hz,1H),4.31–4.21(m,2H),4.1 8–4.06(m,2H),4.01(t,J=9.1Hz,1H),3.89–3.69(m,4H),3.62(dd,J=9.5,3.5Hz,1H),3.40(s,3H),3.36(d,J=6.5Hz,2H). 13 CNMR (101MHz, CDCl3) δ165.4,139.0,138.4,138.0,137.6,133.4,133.3,129. 9,129.8,129.6,129.4,128.6,128.5,128.43,128.41,128.37,128.26,128.2 2,128.1,127.7,127.6,127.5,127.4,98.2,97.7,82.0,80.7,74.9,73.8,73. 5,73.3,73.2,69.5,69.4,69.0,68.7,68.6,67.8,58.4,55.4.HRMS(ESI)calcd for C 55 H 55 N3O 12 [M+NH4] + 967.4124, found 967.4133.

[0123] Preparation of 15p: 15p was prepared from 11f (144.6 mg, 0.21 mmol) and acceptor 14p (66.4 mg, 0.14 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give 15p (134.4 mg, 99%, α / β>20:1)) as a white solid.

[0124] Data: [α] D 21= +165.2 (c 0.31, CHCl3). 1 HNMR (400MHz, CDCl3) δ7.93(t,J=8.5Hz,4H),7.54(t,J=7.5Hz,1H),7.49(t,J=7.5Hz,1H),7.46–7.37(m,6H),7.37– 7.24(m,12H),7.22–7.18(m,2H),7.16–7.08(m,4H),5.88–5.81(m,2H),5.72(d,J=3.5Hz,1H),4.93–4.84(m,2H),4. 78–4.71(m,2H),4.65(d,J=11.5Hz,2H),4.56(d,J=10.9Hz,1H),4.51(d,J=12.1Hz,1H),4.28(t,J=9.4Hz,1H),4.22 (s,2H),3.91–3.79(m,2H),3.76(d,J=9.5Hz,2H),3.70–3.59(m,2H),3.49(dd,J=9.9,5.9Hz,1H),3.37–3.28(m,4H). 13 C NMR (101MHz, CDCl3) δ165.5,165.4,138.2,138.1,137.78,137.76,133.2,129.9,129.81 ,129.76,129.5,128.74,128.66,128.58,128.50,128.47,128.42,128.37,128.31,128.2 ,128.1,128.0,127.9,127.83,127.77,127.6,127.4,127.3,98.1,97.6,79.4,78.6,75.8 ,74.0,73.7,73.3,73.0,70.1,69.3,69.2,68.4,67.9,67.4,58.7,55.1.HRMS(ESI)calcd forC 55 H 55 N3O 12 [M+NH4] + 967.4124, found 967.4129.

[0125] Preparation of 15q: 15q was prepared from 11f (160.7 mg, 0.24 mmol) and acceptor 14q (73.9 mg, 0.16 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1-3:1) to give 15q (135.6 mg, 90%, α / β>20:1) as a white solid.

[0126] Data: [α] D 23 = +192.7 (c 0.33, CHCl3). 1 H NMR (400MHz, CDCl3) δ7.95(d,J=7.9Hz,2H),7.90(d,J=7.9Hz,2H),7.58–7.45(m,4H),7.44–7.22(m ,15H),7.22–7.10(m,7H),5.83(d,J=11.2Hz,1H),5.67(s,1H),5.28(s,1H),5.01(d,J=10.7Hz,2H), 4.88(d,J=10.8Hz,2H),4.69–4.46(m,4H),4.18(s,2H),4.11(t,J=9.5Hz,1H),3.96(d,J=10.5Hz,2 H),3.81(dd,J=22.4,10.5Hz,2H),3.69(d,J=10.3Hz,2H),3.52–3.41(m,4H),3.27(t,J=7.9Hz,1H). 13 CNMR(101MHz,CDCl3)δ165.3,165.2,138.3,138.2,138.0,137.9,133.3,133.2,12 9.8,129.7,129.6,129.4,128.6,128.5,128.4,128.3,128.25,128.17,127.94,127 .90,127.8,127.7,127.6,127.5,127.4,96.5,95.1,80.8,78.4,76.4,75.03,74.96 ,73.6,72.8,70.4,69.1,69.0,68.6,68.1,67.7,60.3,58.2,55.4.HRMS(ESI)calcd forC 55 H 55 N3O 12 [M+NH4] + 967.4124, found 967.4120.

[0127] Preparation of 15r: 15r was prepared from 11f (193.2 mg, 0.29 mmol) and acceptor 14r (19.5 mg, 0.19 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give 15r (98.2 mg, 88%, α / β>20:1) as a white solid.

[0128] Data: [α] D25 = +193.1 (c 0.14, CHCl3). 1 H NMR(400MHz, CDCl3) δ7.98(d,J=7.7Hz,2H),7.88(d,J=7.7Hz,2H),7.61(t,J=7.5Hz,1H), 7.54–7.42(m,3H),7.33(t,J=7.6Hz,2H),7.22(s,5H),5.93(s,1H),5.79(d,J=11.2Hz,1H) ,5.32(s,1H),4.54–4.46(m,2H),4.41(d,J=12.1Hz,1H),4.02–3.90(m,3H),3.83(d,J=11 .2Hz,1H),3.59(d,J=6.4Hz,2H),3.54–3.42(m,2H),2.05–1.93(m,2H),1.85–1.70(m,2H). 13 C NMR (101MHz, CDCl3) δ165.50,165.46,137.6,133.5,133.4,129.92,129.88,129.5,129.3,128.7,128.4 ,127.8,127.6,97.1,74.0,73.6,69.0,68.9,68.7,68.4,65.6,65.5,58.1,33.5,31.8.HRMS(ESI)calcd for C 32 H 33 N3O8[M+Na] + 610.2165, found 610.2164.

[0129] Preparation of 15s: 15s was prepared from 11f (155.4 mg, 0.23 mmol) and acceptor 14s (20.6 mg, 0.15 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1-10:1) to give 15s (87.6 mg, 92%, α / β>20:1) as a white solid.

[0130] Data: [α] D 25 = +230.0 (c 0.15, CHCl3). 1H NMR (400MHz, CDCl3) δ7.98(d,J=7.6Hz,2H),7.86(d,J=7.6Hz,2H),7.59(t,J=7.5Hz ,1H),7.51–7.40(m,3H),7.34–7.13(m,11H),5.90(s,1H),5.73(d,J=11.1Hz,1H),5. 33(s,1H),4.71(t,J=6.1Hz,1H),4.52(d,J=11.9Hz,1H),4.48–4.37(m,2H),3.82(d ,J=11.0Hz,1H),3.68–3.54(m,2H),3.31–3.19(m,3H),3.13(dd,J=16.2,4.8Hz,1H). 13 C NMR (101MHz, CDCl3) δ165.4,140.5,140.2,137.6,133.5,133.3,129.88,129.85,129.5,129.3,128.6,128.5,128.42,12 8.37,127.8,127.7,126.8,124.74,124.71,98.3,80.6,73.6,69.0,68.9,68.7,68.4,58.1,39.9,39.3.HRMS(ESI)calcd for C 36 H 33 N3O7[M+Na] + 642.2216, found 642.2216.

[0131] Preparation of 15t: 15t was prepared from 11f (153.3 mg, 0.23 mmol) and acceptor 14t (27.9 mg, 0.15 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 25:1) to give 15t (93.6 mg, 92%, α / β>20:1) as a white solid.

[0132] Data: [α] D 25 = +261.6 (c 0.11, CHCl3). 1H NMR (400MHz, CDCl3) δ7.98(d,J=7.8Hz,2H),7.88(d,J=7.6Hz,2H),7.57(d, J=8.8Hz,1H),7.53–7.40(m,3H),7.36–7.28(m,2H),7.27–7.13(m,5H),5.9 4(s,1H),5.76(d,J=11.1Hz,1H),5.27(s,1H),4.58–4.34(m,3H),3.96–3.7 7(m,2H),3.59(s,2H),1.83–1.70(m,2H),1.63(s,2H),1.52–1.24(m,18H). 13 C NMR (101MHz, CDCl3) δ165.45,165.41,137.6,133.4,133.3,129.9,129.8,129.5,129.3,128.6,128.3,127.68,127.66,97.0, 76.9,76.8,73.5,69.1,69.0,68.3,58.3,30.0,28.6,24.64,24.55,24.1,23.6,23.5,23.1,22.9,21.3,20.6.HRMS(ESI)calcd forC 39 H 47 N3O7[M+Na] + 692.3312, found 692.3311.

[0133] Preparation of 15u: 15u was prepared from 11f (181 mg, 0.27 mmol) and acceptor 14u (28.0 mg, 0.18 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1-10:1) to give 15u (105.4 mg, 92%, α / β>20:1) as a white solid.

[0134] Data: [α] D 23 = +192.2 (c 0.11, CHCl3). 1H NMR (400MHz, CDCl3) δ7.97(d,J=7.7Hz,2H),7.89(d,J=7.8Hz,2H),7.59(t,J=7.5Hz,1H),7.52–7.40(m,3H),7.32(t,J=7.7Hz,2H),7. 26–7.14(m,5H),5.92(d,J=3.3Hz,1H),5.71(dd,J=11.2,3.3Hz,1H),5.21(d,J=3.6Hz,1H),4.57(t,J=6.3Hz,1H),4.52(d,J=12.0Hz,1 H),4.40(d,J=12.0Hz,1H),3.97(dd,J=11.1,3.6Hz,1H),3.65–3.53(m,2H),3.51–3.43(m,1H),2.42–2.32(m,1H),2.23(d,J=12.3Hz,1 H),1.70–1.59(m,2H),1.37(t,J=11.3Hz,2H),1.14(q,J=11.8Hz,1H),1.05–0.91(m,4H),0.87(d,J=6.5Hz,3H),0.81(d,J=7.0Hz,3H). 13 C NMR (101MHz, CDCl3) δ165.51,165.45,137.7,133.4,133.3,129.89,129.87,129.7,129.5,128.6,128.4,127.7,127.6 ,99.7,81.8,73.6,69.6,69.2,68.6,68.4,59.3,48.8,42.8,34.3,31.9,25.0,22.9,22.3,21.4,16.0.HRMS(ESI)calcd for C 37 H 43 N3O7[M+NH4] + 659.3439, found 659.3446.

[0135] Preparation of 15v: 15v was prepared from 11f (234.7 mg, 0.35 mmol) and acceptor 14v (35.3 mg, 0.23 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to give 15v (136.9 mg, 93%, α / β>20:1) as a white solid.

[0136] Data: [α] D 23 = +183.9 (c 0.33, CHCl3). 1H NMR (400MHz, CDCl3) δ7.98(d,J=7.7Hz,2H),7.89(d,J=7.7Hz,2H),7.58(t,J=7.4Hz,1H),7.51–7.39(m, 3H),7.31(t,J=7.7Hz,2H),7.25–7.13(m,5H),5.94(d,J=3.2Hz,1H),5.83(dd,J=11.2,3.2Hz,1H),5.57( d,J=3.5Hz,1H),4.63(t,J=6.5Hz,1H),4.51(d,J=11.8Hz,1H),4.40(d,J=11.8Hz,1H),3.72(dd,J=11.2 ,3.5Hz,1H),3.59(d,J=6.5Hz,2H),2.17(t,J=3.5Hz,3H),1.93(q,J=11.6Hz,6H),1.65(d,J=3.4Hz,6H). 13 C NMR (101MHz, CDCl3) δ165.49,165.45,137.7,133.4,133.2,129.9,129.8,129.6,129.4,128.6,128. 3,127.61,127.56,91.9,76.1,73.5,69.2,68.9,68.4,67.8,58.1,42.4,36.2,30.7.HRMS(ESI)calcd for C 37 H 39 N3O7[M+NH4] + 655.3126, found 655.3128.

[0137] Preparation of 15w: 15w was prepared from 11f (142.7 mg, 0.21 mmol) and acceptor 14w (53.1 mg, 0.14 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 7:1) to give 15w (121.1 mg, 99%, α / β>20:1) as a white solid.

[0138] Data: [α] D 25 = +161.8 (c 0.10, CHCl3). 1H NMR (400MHz, CDCl3) δ8.20(d,J=7.7Hz,2H),8.00(d,J=7.8Hz,2H),7.89(d,J=7.8Hz,2H),7.60(q,J=6.9Hz,2H),7.53–7.41(m,5H),7. 33(q,J=7.8Hz,3H),7.28–7.14(m,5H),7.00(dd,J=8.5,2.4Hz,1H),6.94(d,J=2.5Hz,1H),5.97(d,J=3.2Hz,1H),5.79(dd,J=11.1,3. 1Hz,1H),5.21(d,J=3.4Hz,1H),4.59–4.50(m,2H),4.44(d,J=11.8Hz,1H),3.89–3.75(m,2H),3.63(q,J=9.8,8.3Hz,2H),2.94–2.83( m,2H),2.35–2.07(m,4H),1.95–1.85(m,1H),1.80–1.66(m,2H),1.63–1.43(m,3H),1.43–1.32(m,2H),1.25–1.17(m,1H),0.93(s,3H). 13 C NMR (101MHz, CDCl3) δ165.49,165.45,148.8,138.3,138.0,137.7,133.53,133.47,13 3.3,130.2,129.9,129.84,129.77,129.5,129.3,128.61,128.58,128.5,128.4,127.9 ,127.8,127.7,127.6,126.6,121.7,118.8,97.3,86.6,73.5,69.2,68.7,68.5,68.4,5 8.3,50.1,44.2,43.0,38.2,37.3,29.6,27.1,26.9,26.1,23.2,12.2.HRMS(ESI)calcd for C 52 H 51 N3O9[M+NH4] + 879.3964, found 879.3972.

[0139] Preparation of 15x: 15x was prepared from 11f (169.1 mg, 0.25 mmol) and acceptor 14x (64.6 mg, 0.17 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to give 15x (136.7 mg, 94%, α / β>20:1) as a yellow solid.

[0140] Data: [α] D 21 = +143.3 (c 0.15, CHCl3). 1 1H NMR (400 MHz, CDCl3) δ 7.98 (d, J = 7.7 Hz, 2H), 7.88 (d, J = 7.8 Hz, 2H), 7.60 (t, J = 7.5 Hz, 1H), 7.53–7.41 (m, 3H), 7.32 (t, J = 7.7 Hz, 2H), 7.26–7.15 (m, 5H), 5.92 (d, J = 3.2 Hz, 1H), 5.78 (dd, J = 11.1, 3.3 Hz, 1H), 5.34–5.28 (m, 2H), 4.56–4.48 (m, 2H), 4.41 (d, J = 11.9 Hz, 1H), 3.79 (dd, J = 11.1, 3.5 Hz, 1H), 3.66–3.54 (m, 3H), 2.46 (d, J = 8.1 Hz, 2H), 2.06–1.93 (m, 3H), 1.92–1.79 (m, 2H), 1.70–1.44 (m, 8H), 1.42–1.28 (m, 4H), 1.21–1.08 (m, 5H), 1.08–0.98 (m, 6H), 0.92 (d, J = 6.4 Hz, 4H), 0.87 (d, J = 6.6 Hz, 6H), 0.69 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 165.5, 140.5, 137.7, 133.5, 133.3, 130.0, 129.9, 129.7, 129.5, 128.7, 128.43, 128.40, 127.8, 127.7, 122.4, 97.4, 79.5, 73.6, 69.3, 69.1, 68.52, 68.47, 58.3, 57.0, 56.4, 50.4, 42.5, 40.2, 40.0, 39.7, 37.2, 36.9, 36.4, 35.9, 32.13, 32.07, 28.4, 28.2, 24.5, 24.0, 23.0, 22.7, 21.2, 19.5, 18.9, 12.0. HRMS (ESI) calcd for C 54 H 69 N3O7 [M+NH4] + 889.5474, found 889.5470.

[0141] Preparation of 15y: 15y was prepared from 11f (155.4 mg, 0.23 mmol) and acceptor 14y (44.3 mg, 0.15 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give 15y (111.7 mg, 94%, α / β>20:1) as a white solid.

[0142] Data: [α] D 22 = +186.7 (c 0.27, CHCl3). 1 H NMR (400MHz, CDCl3) δ7.98(d,J=7.7Hz,2H),7.88(d,J=7.7Hz,2H),7.60(t,J=7.4Hz,1H),7.53–7.42(m,3H),7.32(t, J=7.7Hz,2H),7.28–7.15(m,5H),5.92(d,J=3.3Hz,1H),5.78(dd,J=11.1,3.3Hz,1H),5.32(dd,J=11.1,4.3Hz,2H),4. 57–4.48(m,2H),4.42(d,J=11.8Hz,1H),3.80(dd,J=11.1,3.5Hz,1H),3.67–3.55(m,3H),2.54–2.41(m,3H),2.16–1.9 9(m,3H),1.98–1.82(m,3H),1.73–1.61(m,4H),1.58–1.44(m,2H),1.35–1.28(m,2H),1.14–0.97(m,5H),0.90(s,3H). 13 CNMR(101MHz,CDCl3)δ221.0,165.51,165.48,140.7,137.7,133.5,133.3,12 9.93,129.89,129.6,129.4,128.7,128.5,128.4,127.75,127.72,121.6,97. 3,79.1,73.6,69.2,69.1,68.55,68.52,58.2,51.9,50.4,47.7,40.1,37.1,3 7.0,36.0,31.64,31.60,31.0,28.1,22.0,20.5,19.5,13.7.HRMS(ESI)calcd for C 46 H 51 N3O8[M+NH4] + 791.4014, found 791.4013.

[0143] Preparation of 15z: 15z was prepared from 11f (149.6 mg, 0.22 mmol) and acceptor 14z (69 mg, 0.15 mmol) according to route 1. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to give 15z (121.4 mg, 86%, α / β>20:1) as a white solid.

[0144] Data: [α] D 21 = +141.4 (c 0.28, CHCl3). 1 H NMR(400MHz, CDCl3) δ7.98(d,J=7.7Hz,2H),7.88(d,J=7.7Hz,2H),7.58(t, J=7.5Hz,1H),7.51–7.40(m,3H),7.37–7.28(m,7H),7.23–7.13(m,5H),5.9 3(d,J=3.3Hz,1H),5.79(dd,J=11.0,3.3Hz,1H),5.30(d,J=3.5Hz,1H),5.1 1(d,J=2.9Hz,2H),4.57–4.46(m,2H),4.40(d,J=11.8Hz,1H),3.84(dd,J=11 .1,3.5Hz,1H),3.74–3.65(m,1H),3.59(q,J=9.6,7.9Hz,2H),2.45–2.35(m ,1H),2.33–2.21(m,1H),1.98–1.90(m,2H),1.89–1.76(m,4H),1.66(d,J=12 .5Hz,1H),1.61–1.49(m,2H),1.48–1.32(m,8H),1.30–1.18(m,4H),1.11(q ,J=10.4,9.6Hz,3H),1.04–0.96(m,2H),0.92(d,J=6.6Hz,6H),0.63(s,3H). 13C NMR (101MHz, CDCl3) δ174.1,165.41,165.39,137.6,136.2,133.4,133.2,129.84,129. 81,129.5,129.3,128.6,128.3,128.25,128.18,127.6,127.5,97.2,79.5,73.5,69.2,6 9.1,68.4,68.3,66.1,58.3,56.3,55.9,42.8,42.2,40.4,40.1,35.9,35.4,35.3,34.7, 32.7,31.3,31.0,28.5,28.2,27.3,26.4,24.2,23.5,20.9,18.3,12.1.HRMS(ESI)calcd for C 58 H 69 N3O9[M+NH4] + 969.5372, found 969.5373.

[0145] The above experiments show that when 2-deoxy-2-azidogalactosyl N-phenylacetimidate PTFAI is the donor 11f, the stereoselective alcohol acceptor has a very broad substrate range, including: highly active primary alcohols, secondary alcohols and tertiary alcohols.

[0146] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A preparation method of formula 3, characterized in that:

2. The preparation method according to claim 1, characterized in that: It also includes the preparation method of formula 2, 3. The preparation method according to claim 1 or 2, characterized in that: It also includes the preparation method of formula 4, 4. A method for preparing a compound, characterized in that: Wherein, Lev represents 4-oxopentanoyl, PTFAI represents 5. A method for preparing a compound, characterized in that: OR is selected from the following structures: Among them, STol represents p-tolylthio, MP represents p-methoxyphenyl, and PTFAI represents

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