Method for Synthesizing O-Antigen Saccharide Chain of Helicobacter pylori serotype O:6
20210309683 · 2021-10-07
Assignee
Inventors
Cpc classification
A61P1/04
HUMAN NECESSITIES
C07H15/04
CHEMISTRY; METALLURGY
C08B37/006
CHEMISTRY; METALLURGY
C07H1/00
CHEMISTRY; METALLURGY
C07H15/18
CHEMISTRY; METALLURGY
A61K39/105
HUMAN NECESSITIES
C07K2/00
CHEMISTRY; METALLURGY
International classification
C07H15/04
CHEMISTRY; METALLURGY
C07H1/00
CHEMISTRY; METALLURGY
Abstract
Disclosed is a method for synthesizing an 0-antigen saccharide chain of Helicobacter pylori serotype O:6 using seven glycosylation building blocks. Inexpensive and easily available D-glucosamine, D-galactose, D-mannose and L-fucose are used as starting materials, and seven glycosylation building blocks are obtained through a series of chemical reactions. The saccharide building blocks are then selectively linked together to produce O-antigen oligosaccharide chains of Helicobacter pylori serotype O:6 with different saccharide configuration through a series of glycosylation reactions. Also disclosed is a method to assemble an amino linker at the reducing end of the O-antigen saccharide chain of Helicobacter pylori serotype O:6, and the synthesized oligosaccharide chain with an amino linker can be coupled to a carrier molecule or immobilized on a matrix.
Claims
1. A process of the preparation of compounds according to the following formulas: ##STR00017## ##STR00018## ##STR00019## wherein PG.sub.1, PG.sub.2, PG.sub.3, PG.sub.4, PG.sub.5, PG.sub.6, PG.sub.7, PG.sub.8, PG.sub.9, PG.sub.10, PG.sub.11, PG.sub.12, PG.sub.13, PG.sub.14, PG.sub.15, PG.sub.17, PG.sub.18, PG.sub.19, PG.sub.21, PG.sub.22, PG.sub.23, PG.sub.25, PG.sub.26, PG.sub.27, PG.sub.28, PG.sub.29 and PG.sub.30 are independently selected from any one of hydrogen, acyl, 2-naphthylmethyl and its derivatives, benzyl and its derivatives, allyl and silyl; PG.sub.16 and PG.sub.24 are independently selected from any one of hydrogen, acyl, alkoxycarbonyl and alkoxycarbonyl(acyl); PG.sub.20 is selected from any one of alkanoyl, diformyl, carbobenzyloxy and its derivatives; the leaving groups LG are independently selected from any one of halogen, iminoester group, thio group and phosphonic acid group; wherein the linker is an amino linker —(CH.sub.2).sub.n—N—Y.sub.1Y.sub.2, wherein n=2-40, Y.sub.1 is H or benzyl (Bn), and Y.sub.2 is H or Cbz; and said process comprising: synthesizing the disaccharide compound 9 by coupling a compound 1 as a glycosyl donor and a compound 8 as a glycosyl receptor in an organic solvent following the synthetic route below: ##STR00020##
2. The process of claim 1, further comprising: synthesizing the trisaccharide compound 11 by a deprotection reaction and a coupling reaction following the synthetic route below: ##STR00021## ##STR00022## wherein the deprotection reaction is performed by removing PG.sub.1 protection group on the compound 9 to make a compound 10; and wherein the coupling reaction is performed by coupling a compound 2 as a glycosyl donor and the compound 10 as a glycosyl receptor under Lewis acid catalysis to make the compound 11.
3. The process of claim 2, further comprising: synthesizing the tetrasaccharide compound 13 by coupling a compound 3 as a glycosyl donor and a compound 12 as a glycosyl receptor in an organic solvent to make the compound 13 following the synthetic route below: ##STR00023##
4. The process of claim 3, further comprising: synthesizing the pentasaccharide compound 16 by a first and a second coupling reaction following the synthetic route below: ##STR00024## wherein the first coupling reaction is performed by coupling compound 3 as a glycosyl doner to compound 14 as a glycosyl receptor in an organic solvent to make a compound 15; and wherein the second coupling reaction is performed by coupling the compound 15 as a glycosyl doner to a compound 12 as a glycosyl receptor in an organic solvent to make the compound 16.
5. The process of claim 4, further comprising: synthesizing the octasaccharide 20 compound by a third and a fourth coupling reaction following the synthetic route below: ##STR00025## ##STR00026## wherein the third coupling reaction is performed by coupling a compound 3 as a glycosyl doner to a compound 17 as a glycosyl receptor in an organic solvent to make a compound 18; and wherein the fourth coupling reaction is performed by coupling the compound 18 as a glycosyl doner to a compound 19 as a glycosyl receptor in an organic solvent to make the compound 20.
6. The process of claim 5, further comprising: synthesizing the trisaccharide 24 compound by a fifth and a sixth coupling reaction following the synthetic route below: ##STR00027## wherein the fifth coupling reaction is performed by coupling a compound 6 as a glycosyl doner to a compound 21 as a glycosyl receptor in an organic solvent to make a compound 22; and wherein the sixth coupling reaction is performed by coupling the compound 22 as a glycosyl doner to a compound 23 as a glycosyl receptor in an organic solvent to make the compound 24.
7. The process of claim 6, further comprising synthesizing the tridecasaccharide compound 28 following the synthetic route below: wherein the compound 24 as a glycosyl donor is coupled to a compound 25 as a glycosyl receptor under Lewis acid catalysis to obtain the compound 26; wherein protection groups PG.sub.21 and PG.sub.24 are selectively removed from the compound 26 to obtain a compound 27; and wherein a compound 7 as a glycosyl donor is couple to the compound 27 as a glycosyl receptor under Lewis acid catalysis to make the compound 28.
8. The method of claim 1, wherein the LG is selected from any one of the group consisting of halogen, trichloroacetimidate, N-phenyl trifluoroacetimidate glycoside, ethylthio, phenylthio, p-tolylthio, and dibutylphosphonic acid group.
9. The method of claim 1, wherein the PG.sub.2o is selected from any one of trichloroacetyl, trichloroethoxycarbonyl, phthaloyl, and carbobenzyloxy.
10. The method of claim 1, wherein the PG.sub.16 and PG.sub.24 are selected from any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, levulinyl, 9-fluorenylmethoxycarbonyl, and allyloxycarbonyl.
11. The method of claim 1, wherein the PG.sub.1, PG.sub.9, PG.sub.12, PG.sub.17, PG.sub.21, PG.sub.22 and PG.sub.24 are independently selected from any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl (ClAc), levulinyl, 9-fluorenylmethoxycarbonyl, allyloxycarbonyl, 2-naphthylmethyl, p-methoxybenzyl and allyl.
12. The method of claim 9, wherein the PGI, PG.sub.9, PG.sub.12, PG.sub.17, PG.sub.21, PG.sub.22 and PG.sub.24 are independently selected from any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl (ClAc), levulinyl, 9-fluorenylmethoxycarbonyl, allyloxycarbonyl, 2-naphthylmethyl, p-methoxybenzyl and allyl.
13. The method according to claim 1, wherein the PG.sub.2, PG.sub.3, PG.sub.4, PG.sub.6, PG.sub.7, PG.sub.8, PG.sub.11, PG.sub.13, PG.sub.14, PG.sub.18, PG.sub.25, PG.sub.26, PG.sub.29 and PG.sub.30 are independently selected from any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, allyloxycarbonyl, benzyl, 2-naphthylmethyl, p-methoxybenzyl and allyl.
14. The method of claim 1, wherein the PG.sub.5, PG.sub.10, PG.sub.15, PG.sub.19, PG.sub.23, PG.sub.27 and PG.sub.28 are selected from any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, allyloxycarbonyl, benzyl, 2-naphthylmethyl, p-methoxybenzyl, allyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl and triethylsilyl.
15. The method of claim 13, wherein the PG.sub.5, PG.sub.10, PG.sub.15, PG.sub.19, PG.sub.23, PG.sub.27 and PG.sub.28 are selected from any one of hydrogen, acetyl, benzoyl, pivaloyl, chloroacetyl, allyloxycarbonyl, benzyl, 2-naphthylmethyl, p-methoxybenzyl, allyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl and triethylsilyl.
16. A method for preparing a saccharide-protein conjugate, comprising: reacting the compound 28 of claim 1 with a protein to produce the saccharide-protein conjugate.
Description
BRIEF DESCRIPTION OF THE FIGURES
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DETAILED DESCRIPTION
[0073] Specific embodiments of the invented method are further described in detail with regard to the drawings and the Examples. The specific embodiments are presented here for illustrative purposes only, and are not meant to limit the scope of the invention which is defined by the claims as presented. Various changes and modifications to the disclosed methods known to those skilled in the art shall be covered within the scope of protection.
[0074] If no specific conditions are specified in the examples, they are carried out according to the general conditions or the conditions recommended by the manufacturer. Any reagents or instruments of which no manufacturers are indicated are commercially available conventional products.
[0075] All reagents are analytically pure unless otherwise specified, and have not been further purified unless otherwise specified. All solvents are dried and redistilled by standard methods before use. Unless otherwise noted, all reactions are carried out under the protection of inert gas in dried glassware under magnetic stirring. A silica gel thin plate used for thin layer chromatography (TLC) is of model GF254, produced by Qingdao Haiyang Chemical Co., Ltd. The TLC plate is dyed by ultraviolet light (UV) and a Hanessian solution (cerium sulfate and ammonium molybdate dissolved in a sulfuric acid solution) or a 5% sulfuric acid-ethanol solution, and can be visually detected. Column chromatography silica gel is produced by Qingdao Haiyang Chemical Co., Ltd. (Qingdao, China), and the column chromatography silica gel is of 300-400 meshes. The 1H NMR, 13C NMR, 1H-13C HSQC and 1H-1H COSY spectrograms are measured by NVANCE III 400-MHz, 600-MHz and 700-MHz NMR spectrometers. Unless otherwise specified, CDCl.sub.3 is used as the solvent, TMS (Tetramethylsilane) is used as the internal standard, and the measurement is performed at ambient temperature. Peak type expression methods include: singlet (s), broad singlet (br s), doublet (d), quartet (dd), triplet (t), and multiplet (m). All NMR chemical shifts (6) are recorded in ppm, and the coupling constant (J) is recorded in Hz. A mass spectrum is measured by a Thermo Scientific TSQ Quantum Ultra instrument (Waltham, Mass., USA), and a high resolution mass spectrum is measured by an IonSpec Ultra instrument (Varian, Palo Alto, Calif., U.S).
EXAMPLE 1
Synthesis of saccharide building block 8*
[0076] The synthetic route is shown in
[0077] 2,3-O-propylidene-4-O-benzylmannosethioglycoside was used as a starting material, and after Swern oxidation, the 6-position hydroxyl group was oxidized to aldehyde to obtain compound 1*. Then the carbon chain at position 6 of compound 1* was extended by a Wittig reaction to obtain an olefin compound 2* deoxygenated at position 6. The olefin compound was dihydroxylated under the combined action of potassium osmate (K.sub.2OsO.sub.4), potassium ferricyanate (K.sub.3Fe(CN).sub.6) and potassium carbonate (K.sub.2CO.sub.3) to obtain a 6,7-di-hydroxy compound 3*. The 6,7-di-hydroxyl group was protected by Bn under the action of sodium hydride (NaH) to obtain a compound 4*. After the propylidene group was removed under the action of 80% acetic acid, a compound 5* was obtained, and then under the action of D(+)-10-camphorsulfonic acid (CSA), the 2,3-position hydroxyl group was ring-protected. Ring-opening was performed under a weak acid condition to obtain a 2-OBz protected compound 6*. The 2-OH was protected by Lev to obtain a compound 7*, and then the terminal-position ethylthio group was hydrolyzed by N-iodosuccinimide (NIS) and trifluoromethanesulfonic acid (TfOH). Finally, a reaction was performed with trichloroacetonitrile to obtain a trichloroacetimidate glycosyl donor 8*.
[0078] Specific test operation and steps are as follows:
[0079] Compound 2*: Oxalyl chloride (3.6 mL, 42.3 mmol) was dissolved in dichloromethane (22 mL), and at −78° C., DMSO (6.0 mL, 84.6 mmol) in a dichloromethane solution was added dropwise. After stirring for 15 min, the compound 2,3-O-propylidene-4-O-benzylmannosethioglycoside (10.0 g, 28.2 mmol) in a dichloromethane (115 mL) solution was added to the above reaction solution by a constant pressure dropping funnel. After reaction at −78° C. for 1 h, Et.sub.3N (15.7 mL, 112.8 mmol) was added to the above solution, the reaction temperature was raised to room temperature, and then the reaction was performed at room temperature for 4 h. After it was shown by TLC that the reaction was complete, water was added to quench the reaction. The reaction solution was extracted with dichloromethane, the organic phase was washed with water and a saturated saline solution successively, and then the organic phase was dried with anhydrous Na.sub.2SO.sub.4. The organic phase was concentrated and dried under vacuum to obtain crude aldehyde, and the crude aldehyde was directly used in the next reaction without purification. Methyltriphenylphosphonium bromide (24.2 g, 67.7 mmol) was dissolved in THF (90 mL) at 0° C., and then n-BuLi (23.5 mL, 56.4 mmol, 2.5 M in hexane) was added. The reaction was performed under stirring for 1 h, then the reaction temperature was lowered to −78° C., and THF (28 mL) in which the crude aldehyde product was dissolved was added dropwise. The reaction temperature was raised to room temperature, and the reaction was continued for 12 h. After it was detected by TLC that the reaction was complete, saturated NH.sub.4Cl was added to quench the reaction, and the reaction solution was extracted with ethyl acetate (5×100 mL). The organic phase was dried with anhydrous Na.sub.2SO.sub.4, concentrated, and purified by column chromatography (petroleum ether/ethyl acetate: 100/1.fwdarw.50/1) to obtain the compound 2* (5.5 g, 56%). R.sub.f=0.32, petroleum ether/EtOAc=15:1. [α].sup.25.sub.D=+129.3 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.40-7.23 (m, 5H, arom. H), 5.99 (ddd, J=16.7, 10.6, 5.5 Hz, 1H, 6-H), 5.58 (s, 1H, 1-H), 5.41 (dt, J=17.3, 1.7 Hz, 1H, 7-H), 5.25 (dt, J=10.7, 1.6 Hz, 1H, 7-H′), 4.85 (d, J=11.5 Hz, 1H, Ph—CH.sub.2), 4.63 (d, J=11.5 Hz, 1H, Ph—CH.sub.2), 4.42 (dd, J=10.0, 5.5 Hz, 1H, 5-H), 4.29 (dd, J=7.2, 5.7 Hz, 1H, 3-H), 4.19 (d, J=5.6 Hz, 1H, 2-H), 3.38 (dd, J=10.0, 7.2 Hz, 1H, 4-H), 2.58 (ddq, J=52.5, 13.1, 7.4 Hz, 2H, SCH.sub.2), 1.49 (s, 3H, CH.sub.3), 1.36 (s, 3H, CH.sub.3), 1.28 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ138.2, 135.0, 128.2, 128.0, 127.6, 117.3, 109.4, 80.1, 79.6, 78.5, 76.7, 73.2, 69.5, 28.0, 26.4, 24.4, 14.6. IR (film): v=2985, 2931, 1454, 1380, 1242, 1219, 1162, 1124, 1090, 1066, 996, 872, 748, 697 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.19H.sub.26O.sub.4SNa [M+Na].sup.+373.1449, found 373.1445.
[0080] Compound 3*: Potassium ferricyanide (K.sub.3Fe(CN).sub.6, 46.2 mmol, 15.2 g), potassium osmate dihydrate (K.sub.2O.sub.SO.sub.4.2H.sub.2O, 0.385 mmol, 142 mg) and potassium carbonate (K.sub.2CO.sub.3, 50.8 mmol, 7.0 g) were added to a solution of tert-butanol (77 mL) and water (77 mL).
[0081] Then at 0° C., a solution of compound 2* (5.4 g, 15.4 mmol) in toluene (30 mL) was added dropwise to the reaction solution. The reaction mixture was reacted at 0° C. for 36 h. After it was detected by TLC that the reaction was complete, the reaction was quenched by adding sodium sulfite (Na.sub.2SO.sub.3, 25 g). After stirring for 15 min, the reaction solution was extracted with ethyl acetate. The organic phase was washed with 1 M KOH, dried with anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The organic phase was separated and purified by column chromatography (petroleum ether/ethyl acetate: 5/1-4/1) to obtain the compound 3* (4.3 g, 73%). R.sub.f=0.36, petroleum ether/EtOAc=1:1. [α].sup.25.sub.D=+173.6 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.41-7.28 (m, 5H, arom. H), 5.55 (s, 1H, 1-H), 4.98 (d, J=11.3 Hz, 1H, Ph—CH.sub.2), 4.64 (d, J=11.3 Hz, 1H, Ph—CH.sub.2), 4.32 (dd, J=6.9, 5.7 Hz, 1H, 3-H), 4.20 (dd, J=5.7, 0.7 Hz, 1H, 2-H), 4.06 (dd, J=9.9, 6.3 Hz, 1H, 5-H), 3.89 (ddt, J=6.8, 4.8, 2.2 Hz, 1H, 6-H), 3.69 (dd, J=10.0, 7.0 Hz, 1H, 4-H), 3.66-3.63 (m, 2H, 7-H/7-H′), 3.54 (d, J=2.8 Hz, 1H, OH), 2.63 (ddq, J=51.1, 12.9, 7.4 Hz, 2H, SCH.sub.2), 1.54 (s, 3H, CH.sub.3), 1.37 (s, 3H, CH.sub.3), 1.29 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ137.2, 109.6, 79.7, 79.5, 78.3, 76.5, 73.2, 72.8, 67.7, 62.8, 28.1, 26.4, 24.1, 14.2. IR (film): v=3446, 2984, 2931, 1454, 1380, 1242, 1219, 1162, 1066, 872, 750, 699 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.19H.sub.28O.sub.6SNa [M+Na].sup.+407.1504, found 407.1507.
[0082] Compound 4*: The compound 3* (1.7 g, 4.4 mmol) was dissolved in DMF (22 mL), and sodium hydride (0.7 g, 17.7 mmol) (60% dispersed in mineral oil) was added. The reaction temperature was lowered to 0° C., and then BnBr (2.1 mL, 17.7 mmol) was added. The reaction was performed under stirring at room temperature for 3 h. After it was detected by TLC that the reaction was complete, the reaction was quenched by adding an appropriate amount of methanol. The reaction solution was extracted with dichloromethane, and the organic phase was sequentially washed with water and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, and concentrated under reduced pressure. The crude product was separated and purified by column chromatography (petroleum ether/ethyl acetate: 100/1.fwdarw.50/1) to obtain the compound 4* (2.2 g, 89%). R.sub.f=0.27, petroleum ether/EtOAc=20:1. [α].sup.25.sub.D=+117.7 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.40-7.17 (m, 15H, arom. H), 5.54 (s, 1H, 1-H), 4.83 (d, J=11.4 Hz, 1H,) Ph—CH.sub.2, 4.72 (d, J=11.8 Hz, 1H, Ph—CH.sub.2), 4.67 (d, J=11.8 Hz, 1H, Ph—CH.sub.2), 4.53 (d, J=11.4 Hz, 1H, Ph—CH.sub.2), 4.49 (d, J=12.0 Hz, 1H, Ph—CH.sub.2), 4.43 (d, J=12.0 Hz, 1H, Ph—CH.sub.2), 4.32-4.24 (m, 2H, 3-H/5-H), 4.15 (d, J=5.7 Hz, 1H, 2-H), 4.04 (td, J=5.7, 1.5 Hz, 1H, 6-H), 3.68 (d, J=5.7 Hz, 2H, 7-H/7-H′), 3.68 (dd, J=10.0, 7.0 Hz, 1H, 4-H), 2.59 (ddq, J=67.5, 12.8, 7.4 Hz, 2H, SCH.sub.2), 1.46 (s, 3H, CH.sub.3), 1.35 (s, 3H, CH.sub.3), 1.23 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ138.7, 138.5, 138.2, 128.3, 128.2, 128.2, 128.0, 127.7, 127.5, 127.5, 127.4, 127.3, 109.3, 79.5, 79.0, 77.9, 76.4, 76.3, 73.2, 72.7, 72.5, 70.5, 69.3, 28.0, 26.5, 23.8, 14.2. IR (film): v=2929, 1453, 1380, 1218, 1093, 1066, 1027, 870, 734, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.33H.sub.30O.sub.6SNa [M+Na].sup.+587.2443, found 587.2429.
[0083] Compound 5*: The compound 4* (2.2 g, 3.8 mmol) was dissolved in an 80% acetic acid solution (40 mL), and the reaction mixture was reacted at 60° C. for 5 h. After it was detected by TLC that the reaction was complete, the reaction solution was concentrated by rotary evaporation, and an appropriate amount of DCM was added for dissolution. Then the organic phase was sequentially washed with saturated NaHCO.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The organic phase was separated and purified by column chromatography (petroleum ether/ethyl acetate: 4/1) to obtain the compound 5* (2 g, quan.). R.sub.f=0.33, petroleum ether/EtOAc=2:1. [α].sup.25.sub.D=+119.6 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.45-7.14 (m, 15H, arom. H), 5.25 (d, J=1.8 Hz, 1H, 1-H), 4.74 (d, J=11.9 Hz, 1H, Ph—CH.sub.2), 4.69 (d, J=11.8 Hz, 1H, Ph—CH.sub.2), 4.68 (d, J=11.7 Hz, 1H, Ph—CH.sub.2), 4.63 (d, J=11.5 Hz, 1H, Ph—CH.sub.2), 4.53 (d, J=12.0 Hz, 1H, Ph—CH.sub.2), 4.48 (d, J=11.9 Hz, 1H, Ph—CH.sub.2), 4.28 (dd, J=9.5, 1.6 Hz, 1H, 5-H), 4.00 (ddd, J=6.7, 5.1, 1.6 Hz, 1H, 6-H), 3.93 (ddd, J=7.4, 3.9, 2.2 Hz, 1H, 2-H), 3.89 (dq, J=5.7, 3.4, 2.8 Hz, 1H, 3-H), 3.81 (d, J=5.2 Hz, OH), 3.79 (d, J=10.0, 5.2 Hz, 1H, 7-H), 3.77 (dd, J=10.0, 7.0 Hz, 1H, 4-H), 3.70 (dd, J=10.2, 6.7 Hz, 1H, 7-H′), 2.71-2.50 (m, 2H, SCH.sub.2), 2.47 (d, J=4.6 Hz, 1H, 2-OH), 2.28 (d, J=5.7 Hz, 1H, 3-OH), 1.25 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ138.6, 138.3, 138.2, 128.6, 128.4, 128.3, 127.9, 127.9, 127.8, 127.7, 127.6, 127.5, 83.4, 77.8, 76.4, 74.2, 73.4, 72.6, 72.5, 72.2, 71.7, 70.6, 24.7, 14.7. IR (film): v=3420, 2924, 1453, 1075, 1027, 792, 733, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.30H.sub.36O.sub.6SNa [M+Na].sup.+547.2130, found 547.2118.
[0084] Compound 6*: The compound 5* (1.06 g, 2.0 mmol) was dissolved in anhydrous DCM (20 mL), and triethyl orthobenzoate (0.7 mL, 3.0 mmol) and CSA (23 mg, 0.1 mmol) were added. The reaction was performed under stirring at room temperature for 1 h. After it was detected by TLC that the raw materials were completely converted into intermediates, water (70 μL, ˜4.0 mmol) was added. The reaction was performed under stirring at room temperature for 1 h. After it was detected by TLC that the reaction was complete, an appropriate amount of DCM was added for dilution. The organic phase was washed with saturated NaHCO.sub.3, and the aqueous layer was extracted once with DCM. The organic phases were combined and washed with a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, filtered, concentrated, and separated and purified by column chromatography (petroleum ether/ethyl acetate: 15/1.fwdarw.10/1) to obtain the compound 6* (1.0 g, 80%). R.sub.f=0.43, petroleum ether/EtOAc=4:1. [α].sup.25.sub.D=+47.5 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ8.31-7.12 (m, 20H, arom. H), 5.42 (dd, J=3.3, 1.7 Hz, 1H, 2-H), 5.39 (d, J=1.6 Hz, 1H, 1-H), 4.82 (t, J=11.4 Hz, 2H, Ph—CH.sub.2), 4.71 (dd, J=12.8, 11.5 Hz, 2H, Ph—CH.sub.2), 4.53 (s, 2H, Ph—CH.sub.2), 4.38-4.32 (m, 1H, 5-H), 4.20 (ddd, J=8.9, 5.4, 3.3 Hz, 1H, 3-H), 4.04 (t, J=9.4 Hz, 1H, 4-H), 4.04 (m, 1H, 6-H), 3.83 (dd, J=10.3, 5.0 Hz, 1H, 7-H), 3.74 (dd, J=10.3, 6.8 Hz, 1H, 7-H′), 2.76-2.54 (m, 2H, SCH.sub.2), 2.08 (d, J=5.4 Hz, 1H, 3-OH), 1.27 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ166.1, 138.8, 138.4, 138.2, 133.3, 129.8, 129.7, 128.5, 128.3, 128.3, 128.0, 127.8, 127.5, 127.5, 127.4, 82.1, 78.8, 76.4, 74.8, 74.7, 73.3, 72.7, 72.2, 71.8, 71.1, 25.4, 14.9. IR (film): v=3435, 3030, 2870, 1719, 1452, 1267, 1089, 1026, 902, 735, 711, 697 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.37H.sub.40O.sub.7SNa [M+Na].sup.+651.2392, found 651.2383.
[0085] Compound 7*: The compound 6* (1.6 g, 2.5 mmol) was dissolved in anhydrous DCM (20 mL). Then levulinic acid (0.4 mL, 3.8 mmol), N,N-dicyclohexyl diimide (0.79 g, 3.8 mmol) and 4-dimethylaminopyridine (0.47 g, 3.8 mmol) were added in sequence. The reaction was performed under stirring at room temperature for 1 h. After it was detected by TLC that the reaction was complete, an appropriate amount of DCM was added for dilution. The organic layer was washed sequentially with saturated NaHCO.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, filtered, concentrated, and separated and purified by column chromatography (petroleum ether/ethyl acetate: 10/1.fwdarw.8/1) to obtain the compound 7* (1.5 g, 82%). R.sub.f=0.43, petroleum ether/EtOAc=3:1. [α].sup.25.sub.D=+27.9 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ8.15-7.10 (m, 20H, arom. H), 5.58 (dd, J=3.2, 1.7 Hz, 1H, 2-H), 5.41 (dd, J=9.6, 3.2 Hz, 1H, 3-H), 5.37 (d, J=1.6 Hz, 1H, 1-H), 4.87 (d, J=12.0 Hz, 1H, Ph—CH.sub.2), 4.75 (d, J=11.9 Hz, 1H, Ph—CH.sub.2), 4.68 (d, J=11.1 Hz, 1H, Ph—CH.sub.2), 4.62 (d, J=11.1 Hz, 1H, Ph—CH.sub.2), 4.51 (s, 2H, Ph—CH.sub.2), 4.45 (d, J=9.9 Hz, 1H, 5-H), 4.24 (t, J=9.7 Hz, 1H, 4-H), 4.04 (ddd, J=6.4, 5.0, 1.2 Hz, 1H, 6-H), 3.80 (dd, J=10.3, 5.0 Hz, 1H, 7-H), 3.72 (dd, J=10.3, 6.8 Hz, 1H, 7-H′), 2.77-2.53 (m, 4H, SCH.sub.2/CH.sub.2), 2.51-2.33 (m, 2H, CH.sub.2), 2.07 (s, 3H, Ac), 1.27 (t, J=7.4 Hz, 3H, SCH.sub.2CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ206.1, 171.7, 165.4, 138.7, 138.3, 138.0, 133.4, 129.8, 129.6, 128.5, 128.3, 128.3, 127.8, 127.6, 127.5, 127.5, 127.4, 127.4, 82.0, 78.9, 74.6, 73.5, 73.4, 73.3, 72.8, 72.4, 72.1, 71.1, 37.8, 29.7, 27.9, 25.2, 14.8. IR (film): v=2928, 1719, 1452, 1265, 1151, 1089, 1026, 736, 711, 697 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.42H.sub.46O.sub.9SNa [M+Na].sup.+749.2760, found 749.2763.
[0086] Compound 8*: The compound 7* (1.34 g, 1.84 mmol) was dissolved in CH.sub.2Cl.sub.2 (18 mL), and then water (0.33 mL, 18.4 mmol) was added and stirred. NIS (0.62 g, 2.76 mmol) and TfOH (36 μL, 0.41 mmol) were added at 0° C., and then stirred at 0° C. for 1.5 h. After it was detected by TLC that the reaction was complete, Et.sub.3N was added to stop the reaction, and an appropriate amount of DCM was added for dilution. Then the organic phase was washed with 10% Na.sub.2S.sub.2O.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, filtered, concentrated, and separated and purified by column chromatography (petroleum ether/ethyl acetate: 3/1.fwdarw.2/1) to obtain the corresponding hemiacetal (1.33 g, quan.). R.sub.f=0.36, petroleum ether/EtOAc=1:1.
[0087] The obtained hemiacetal (238 mg, 0.35 mmol) was dissolved in CH.sub.2Cl.sub.2 (4 mL), and CCl.sub.3CN (107 μL, 1.07 mmol) and DBU (7 μL, 0.046 mmol) were added at 0° C. The reaction was performed under stirring at room temperature for 45 min. After it was detected by TLC that the reaction was complete, the reaction solution was concentrated at 30° C. Then the reaction solution was separated and purified by silicagel column chromatography (petroleum ether/EtOAc: 6/1.fwdarw.4/1) to obtain the compound 8* (266 mg, 92%). R.sub.f=0.33, petroleum ether/EtOAc=3:1. .sup.1H NMR (400 MHz, Chloroform-d) δ8.72 (s, 1H, arom. H), 8.13-7.93 (m, 2H, arom. H), 7.71-7.53 (m, 1H, arom. H), 7.44-7.14 (m, 16H, arom. H), 6.40 (d, J=2.1 Hz, 1H, 1-H), 5.70 (dd, J=3.3, 2.1 Hz, 1H, 2-H), 5.54 (dd, J=9.4, 3.3 Hz, 1H, 3-H), 4.89 (d, J=11.9 Hz, 1H, Ph—CH.sub.2), 4.76 (d, J=11.9 Hz, 1H, Ph—CH.sub.2), 4.72 (d, J=10.9 Hz, 1H, Ph—CH.sub.2), 4.64 (d, J=10.9 Hz, 1H, Ph—CH.sub.2), 4.47 (d, J=1.8 Hz, 2H, Ph—CH.sub.2), 4.37 (t, J=9.7 Hz, 1H, 4-H), 4.30 (d, J=10.0 Hz, 1H, 5-H), 4.10 (t, J=6.4 Hz, 1H, 6-H), 3.76 (dd, J=10.2, 5.7 Hz, 1H, 7-H), 3.71 (dd, J=10.1, 6.8 Hz, 1H, 7′-H), 2.74 (dt, J=18.5, 7.2 Hz, 1H, CH.sub.2), 2.62 (dt, J=18.5, 6.4 Hz, 1H, CH.sub.2), 2.54-2.36 (m, 2H, CH.sub.2), 2.09 (s, 3H, CH.sub.3CO). .sup.13C NMR (101 MHz, Chloroform-d) δ206.0, 171.8, 165.2, 160.1, 138.7, 138.2, 137.7, 133.6, 129.8, 129.2, 128.6, 128.3, 128.3, 128.0, 127.7, 127.6, 127.5, 127.4, 127.4, 94.9, 90.7, 78.8, 74.9, 74.7, 73.3, 73.1, 72.6, 72.6, 70.8, 68.7, 37.8, 29.7, 27.9.
EXAMPLE 2
Synthesis of saccharide building block 13*
[0088] The synthetic route is shown in
[0089] As shown in
[0090] The saccharide building block 13* was synthesized as follows: first the previously prepared intermediate compounds 3 and 4 were used as starting materials; under the action of dibutyltin oxide (Bu.sub.2SnO), the 3-OH of the compound 5* was selectively protected by Bn to obtain a compound 12*; and finally the 2-OH was protected by an acetyl group to obtain the heptose building block 13*.
[0091] The experimental procedure is as follows:
[0092] Compound 9*: The compound 3* (0.77 g, 2 mmol) and Bu.sub.2SnO (0.75 g, 3 mmol) were dissolved in dry toluene (10 mL), and reflux reaction was performed for 4 h. In the process, the toluene-water azeotropic mixture (˜5 mL) was removed by a Dean-Stark device, and then the reaction system was cooled to room temperature, concentrated and dried in vacuum. The above residue was dissolved in CH.sub.3CN (5 mL), and then CsF (456 mg, 3 mmol) and BnBr (360 μL, 3 mmol) were added. The reaction was performed under stirring at 70° C. for 10 h. After it was detected by TLC that the reaction was complete, the reaction mixture was filtered with celite and concentrated. The crude product was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate: 8/1) to obtain the compound 9* (0.63 g, 66%). R.sub.f=0.56, petroleum ether/EtOAc=3:1. [α].sup.25.sub.D=+124.6 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.48-7.21 (m, 10H, arom. H), 5.52 (s, 1H, 1-H), 4.92 (d, J=11.4 Hz, 1H, Ph—CH.sub.2), 4.59 (d, J=11.3 Hz, 1H, Ph—CH.sub.2), 4.53 (d, J=12.0 Hz, 1H, Ph—CH.sub.2), 4.49 (d, J=12.0 Hz, 1H, Ph—CH.sub.2), 4.34-4.27 (m, 1H, 3-H), 4.18 (dd, J=5.7, 0.8 Hz, 1H, 2-H), 4.13-4.04 (m, 2H, 6-H/5-H), 3.69 (dd, J=9.4, 6.9 Hz, 1H, 4-H), 3.58 (dd, J=10.3, 6.5 Hz, 1H, 7-H), 3.54 (dd, J=10.4, 3.5 Hz, 1H, 7-H′), 3.00 (d, J=2.7 Hz, 1H, 6-OH), 2.57 (ddq, J=57.7, 12.8, 7.4 Hz, 2H, SCH.sub.2), 1.52 (s, 3H, CH.sub.3), 1.36 (s, 3H, CH.sub.3), 1.24 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ138.2, 137.7, 128.4, 128.3, 128.2, 127.8, 127.7, 127.6, 109.5, 79.6, 78.6, 77.9, 76.5, 73.5, 72.8, 72.3, 70.9, 68.5, 28.0, 26.4, 24.1, 14.2. IR (film): v=3482, 2984, 2930, 1454, 1380, 1241, 1219, 1162, 1068, 1027, 870, 736, 698 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.26H.sub.34O.sub.6SNa [M+Na].sup.+497.1974, found 497.1969.
[0093] Compound 10*: The compound 9* (567 mg, 1.2 mmol) was dissolved in dry CH.sub.2Cl.sub.2 (23 mL), and then LevOH (185 μL, 1.8 mmol), DCC (370 mg, 1.8 mmol) and DMAP (220 mg, 1.86 mmol) were added. The reaction was performed under stirring at room temperature for 1 h. After it was detected by TLC that the reaction was complete, an appropriate amount of DCM was added for dilution. The reaction solution was washed with saturated NaHCO.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether/ethyl acetate: 8/1.fwdarw.4/1) to obtain the compound 10* (707 mg, quan.). R.sub.f=0.32, petroleum ether/EtOAc=4:1. [α].sup.25.sub.D=+105.8 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.45-7.16 (m, 10H, arom. H), 5.57-5.52 (m, 1H, 6-H), 5.52 (s, 1H, 1-H), 4.87 (d, J=11.6 Hz, 1H, Ph—CH.sub.2), 4.55 (d, J=11.6 Hz, 1H, Ph—CH.sub.2), 4.47 (d, J=12.0 Hz, 1H, Ph—CH.sub.2), 4.43 (d, J=12.0 Hz, 1H, Ph—CH.sub.2), 4.26 (t, J=6.3 Hz, 1H, 3-H), 4.19 (dd, J=10.3, 2.2 Hz, 1H, 5-H), 4.13 (dd, J=5.7, 0.7 Hz, 1H, 2-H), 3.68-3.58 (m, 3H, 4-H/7-H/7-H′), 2.75-2.63 (m, 3H, CH.sub.2), 2.62-2.44 (m, 3H, SCH.sub.2/CH.sub.2), 2.16 (s, 3H, OAc), 1.46 (s, 3H, CH.sub.3), 1.34 (s, 3H, CH.sub.3), 1.28 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ206.4, 171.8, 138.1, 138.1, 128.3, 128.0, 127.6, 127.5, 127.5, 109.4, 79.5, 78.8, 76.4, 76.2, 73.0, 72.5, 71.6, 68.9, 68.0, 37.9, 29.9, 28.0, 28.0, 26.4, 23.9, 14.4. IR (film): v=2984, 2931, 1739, 1719, 1361, 1218, 1159, 1096, 1066, 871, 748, 698 cm.sup.−1. HRMS (ESI) nilz calcd for C.sub.31H.sub.40O.sub.8SNa [M+Na].sup.+595.2342, found 595.2331.
[0094] Compound 11*: The compound 10* (652 mg, 1.14 mmol) was dissolved in a 80% acetic acid solution (11 mL), and the reaction mixture was reacted at 60° C. for 5 h. After it was detected by TLC that the reaction was complete, the reaction solution was concentrated by rotary evaporation. An appropriate amount of DCM was added for dissolution, and then the reaction solution was washed with saturated NaHCO.sub.3 and a saturated saline solution sequentially, dried with anhydrous Na.sub.2SO.sub.4, filtered, concentrated, and dried in vacuum. The above residue was dissolved in pyridine (4 mL), then Ac.sub.2O (1.1 mL, 11.4 mmol) and DMAP (cat.) were added, and the reaction was performed under stirring at room temperature for 3 h. After it was detected by TLC that the reaction was complete, the reaction mixture was evaporated to dryness, and an appropriate amount of DCM was added for dilution. The reaction solution was sequentially washed with 1 M HCl (aq), a saturated NaHCO.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, concentrated, and separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate: 10/16/1) to obtain the compound 11* (605 mg, 86%). R.sub.f=0.35, petroleum ether/EtOAc=3:1. [α].sup.25.sub.D=+79.9 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.37-7.20 (m, 10H, arom. H), 5.51 (td, J=6.2, 2.1 Hz, 1H, 6-H), 5.30 (dd, J=3.3, 1.8 Hz, 1H, 2-H), 5.25 (dd, J=9.3, 3.3 Hz, 1H, 3-H), 5.21 (d, J=1.7 Hz, 1H, 1-H), 4.64 (s, 2H, Ph—CH.sub.2), 4.51 (d, J=2.6 Hz, 2H, Ph—CH.sub.2), 4.33 (dd, J=9.9, 2.1 Hz, 1H, 5-H), 4.01 (t, J=9.6 Hz, 1H, 4-H), 3.77 (dd, J=10.3, 5.8 Hz, 1H, 7-H), 3.62 (dd, J=10.2, 6.5 Hz, 1H, 7-H′), 2.74 (t, J=6.7 Hz, 2H, CH.sub.2), 2.69-2.52 (m, 4H, SCH.sub.2/CH.sub.2), 2.17 (s, 3H, OAc), 2.11 (s, 3H, OAc), 1.92 (s, 3H, OAc), 1.27 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ206.3, 171.7, 169.9, 169.6, 138.0, 137.9, 128.4, 128.4, 127.7, 127.6, 127.6, 127.5, 81.8, 74.3, 73.8, 73.2, 72.3, 71.6, 71.5, 68.3, 37.9, 29.8, 28.0, 25.1, 20.9, 20.8, 14.8. IR (film): v=2922, 1744, 1719, 1365, 1236, 1157, 1093, 914, 734, 698 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.32H.sub.40O.sub.10SNa [M+Na].sup.+639.2240, found 639.2229.
[0095] Compound 12*: The compound 5* (600 mg, 1.14 mmol) and Bu.sub.2SnO (426 mg, 1.71 mmol) were dissolved in dry toluene (5.7 mL), and reflux reaction was performed for 4 h. In the process, the toluene-water azeotropic mixture (˜3 mL) was removed by a Dean-Stark device, and then the reaction system was cooled to room temperature, concentrated and dried in vacuum. The above residue was dissolved in CH.sub.3CN (3 mL), then CsF (260 mg, 1.71 mmol) and BnBr (200 μL, 1.71 mmol) were added, and the reaction was performed under stirring at 70° C. for 10 h. After it was detected by TLC that the reaction was complete, the reaction mixture was filtered with celite and concentrated. The crude product was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate: 6/1) to obtain the compound 12* (540 mg, 77%). R.sub.f=0.38, petroleum ether/EtOAc=3:1. [α].sup.25.sub.D=+95.5 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.43-7.12 (m, 20H, arom. H), 5.33 (d, J=1.5 Hz, 1H, 1-H), 4.79 (d, J=10.9 Hz, 1H, Ph—CH.sub.2), 4.75 (d, J=12.0 Hz, 1H, Ph—CH.sub.2), 4.68 (d, J=12.0 Hz, 1H, Ph—CH.sub.2), 4.65-4.63 (m, 2H, Ph—CH.sub.2), 4.59 (d, J=10.9 Hz, 1H, Ph—CH.sub.2), 4.50 (d, J=12.0 Hz, 1H, Ph-CH.sub.2), 4.45 (d, J=12.1 Hz, 1H, Ph—CH.sub.2), 4.30 (dd, J=9.5, 1.3 Hz, 1H, 5-H), 4.05 (dt, J=3.4, 1.7 Hz, 1H, 2-H), 3.99 (ddd, J=6.4, 4.7, 1.5 Hz, 1H, 6-H), 3.90 (t, J=9.2 Hz, 1H, 4-H), 3.85 (dd, J=8.8, 3.1 Hz, 1H, 3-H), 3.76 (dd, J=10.4, 4.6 Hz, 1H, 7-H), 3.69 (dd, J=10.5, 6.9 Hz, 1H, 7-H′), 2.72-2.47 (m, 2H, SCH.sub.2), 2.58 (d, J=2.2 Hz, 1H, 2-OH), 1.24 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ138.7, 138.4, 138.3, 137.6, 128.6, 128.3, 128.3, 128.2, 128.1, 128.0, 127.8, 127.7, 127.6, 127.6, 127.4, 127.3, 83.0, 80.9, 78.1, 74.7, 74.6, 73.2, 72.3, 72.1, 71.9, 70.8, 69.6, 29.7, 24.6, 14.7. IR (film): v=2917, 2849, 1453, 1088, 1027, 790, 733, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.37H.sub.42O.sub.6SNa [M+Na].sup.+637.2600, found 637.2585.
[0096] Compound 13*: The compound 12* (1.2 g, 2 mmol) was dissolved in pyridine (6 mL), then Ac2O (1.1 mL, 11.4 mmol) and DMAP (cat.) were added, and the reaction was performed under stirring at room temperature for 3 h. After it was detected by TLC that the reaction was complete, the reaction mixture was evaporated to dryness, and an appropriate amount of DCM was added for dilution. The reaction solution was sequentially washed with 1 M HCl (aq), a saturated NaHCO.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, concentrated, and separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate: 10/1.fwdarw.5/1) to obtain the compound 13* (1.2 g, 93%). R.sub.f=0.36, petroleum ether/EtOAc=3:1. [a].sup.25.sub.D=+67.4 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.48-7.11 (m, 20H, arom. H), 5.39 (dd, J=3.0, 1.7 Hz, 1H, 2-H), 5.25 (d, J=1.7 Hz, 1H, 1-H), 5.25 (d, J=1.7 Hz, 1H, 1-H, Ph—CH.sub.2), 4.84 (d, J=10.8 Hz, 1H, Ph—CH.sub.2), 4.77 (d, J=11.9 Hz, 1H, Ph—CH.sub.2), 4.68 (d, J=11.90 Hz, 1H, Ph—CH.sub.2), 4.66 (d, J=11.19 Hz, 1H, Ph—CH.sub.2), 4.57 (d, J=10.8 Hz, 1H, Ph—CH.sub.2), 4.50 (d, J=11.8 Hz, 1H, Ph—CH.sub.2), 4.50 (d, J=11.1 Hz, 1H, Ph—CH.sub.2), 4.46 (d, J=12.1 Hz, 1H, Ph—CH.sub.2), 4.29 (dd, J=9.3, 1.2 Hz, 1H, 5-H), 3.99 (ddd, J=6.4, 4.7, 1.5 Hz, 1H, 6-H), 3.97 (t, J=9.3 Hz, 1H, 4-H), 3.92 (dd, J=9.1, 3.0 Hz, 1H, 3-H), 3.75 (dd, J=10.4, 4.6 Hz, 1H, 7-H), 3.68 (dd, J=10.4, 6.9 Hz, 1H, 7-H′), 2.72-2.50 (m, 2H, SCH.sub.2), 2.10 (s, 3H, OAc), 1.25 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ170.2, 138.8, 138.8, 138.4, 137.6, 128.4, 128.3, 128.2, 128.2, 127.8, 127.8, 127.5, 127.5, 127.4, 82.1, 79.0, 78.6, 74.8, 74.6, 73.3, 72.4, 72.3, 71.8, 71.1, 70.4, 25.3, 21.1, 14.8. IR (film): v=3029, 2869, 1742, 1453, 1369, 1230, 1091, 1027, 734, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.39H.sub.44O.sub.7O.sub.7SNa [M+Na].sup.+679.2705, found 679.2694.
EXAMPLE 3
Synthesis of reducing-end trisaccharide
[0097] The synthetic route of reducing-end trisaccharide is shown in
[0098] 3.1 The glycosylation reaction conditions of the reducing-end trisaccharide shown in
TABLE-US-00001 TABLE 1 Optimization of glycosylation reaction conditions
[0099] 3.2 The conditions for removing the acetyl group are as follows: the starting material was dissolved in MeOH/THF (v/v, 1:1), the reaction concentration was 0.05 M, and 0.5 equivalent of MeONa (5M in MeOH) was added. The reaction temperature was room temperature. After it was detected by TLC that the reaction was complete, the reaction solution was neutralized with Amerlite IR 120 (H±) resin to reach a pH of 7. The reaction solution was filtered, concentrated, and separated and purified by silica gel column chromatography.
[0100] The experimental procedure is as follows:
[0101] Compound 14*: According to reaction conditions 3.1, the glycosyl donor 13* (980 mg, 1.49 mmol) and the glycosyl receptor linker (1.04 g, 3.43 mmol) reacted to obtain the compound 14* (1.13 g, 85%). R.sub.f=0.33, petroleum ether/EtOAc=4:1. [α].sup.25.sub.D=+14.8 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.46-7.11 (m, 30H, arom. H), 5.27 (s, 1H, 2-H), 5.17 (s, 2H, Ph—CH.sub.2), 4.84 (d, J=10.7 Hz, 1H, Ph—CH.sub.2), 4.78-4.59 (m, 3H, Ph—CH.sub.2), 4.68 (s, 1H, 1-H), 4.56 (d, J=10.8 Hz, 1H, Ph—CH.sub.2), 4.53-4.37 (m, 5H), 3.98 (t, J=5.8 Hz, 1H, 6-H), 3.94-3.78 (m, 3H, 3-H/4-H/-5-H), 3.73 (dd, J=10.4, 4.7 Hz, 1H, 7-H), 3.67 (dd, J=10.4, 6.7 Hz, 1H, 7-H′), 3.74-3.56 (m, 1H, CH.sub.2), 3.44-3.20 (m, 3H, CH.sub.2), 2.09 (s, 3H, CH.sub.3CO), 1.89-1.64 (m, 2H, CH.sub.2). .sup.13C NMR (101 MHz, Chloroform-d) δ170.2, 156.4 (d, J=52.1 Hz), 138.7, 138.4 (d, J=3.1 Hz), 137.8, 137.8, 136.8, 128.6, 128.4, 128.3, 128.2, 128.1, 127.9, 127.9, 127.7, 127.5, 127.4, 97.5, 78.5, 78.4, 74.8, 74.3, 73.3, 72.5, 72.2, 71.8, 70.8, 68.7, 67.2, 65.3 (d, J=22.4 Hz), 50.7 (d, J=24.7 Hz), 44.1 (d, J=91.4 Hz), 27.9 (d, J=48.9 Hz), 21.0. IR (film): v=3030, 2919, 1744, 1698, 1453, 1368, 1232 1090, 1027, 734, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.55H.sub.59O.sub.10NNa [M+Na].sup.+916.4037, found 916.4020.
[0102] Compound 15*: According to reaction conditions 3.2, the ester group of the compound 14* (960 mg, 1.12 mmol) was removed to obtain the compound 15* (908 mg, quan.). R.sub.f=0.34, petroleum ether/EtOAc=2:1. [α].sup.25.sub.D=+26.8 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.46-7.08 (m, 30H, arom. H), 5.23-5.11 (m, 2H, Ph—CH.sub.2) 4.79 (d, J=10.8 Hz, 1H, Ph—CH.sub.2), 4.75-4.60 (m, 3H, Ph—CH.sub.2), 4.66 (s, 1H, 1-H), 4.58 (d, J=10.8 Hz, 1H, Ph-CH.sub.2), 4.51-4.39 (m, 5H, Ph—CH.sub.2), 4.02-3.93 (m, 1H, 6-H), 3.89 (s, 1H, 2-H), 3.88-3.77 (m, 3H, 3-H/4-H/5-H), 3.74 (dd, J=10.4, 4.7 Hz, 1H, 7-H), 3.68 (dd, J=10.4, 6.7 Hz, 1H, 7-H′), 3.74-3.58 (m, 1H, CH.sub.2), 3.45-3.11 (m, 3H, CH.sub.2), 2.35 (d, J=13.5 Hz, 1H, 2-OH), 1.88-1.62 (m, 2H, CH.sub.2). .sup.13C NMR (101 MHz, Chloroform-d) δ156.4 (d, J=47.1 Hz), 138.7, 138.4, 138.4, 137.9, 137.8, 136.8, 128.6, 128.5, 128.5, 128.3, 128.3, 128.2, 127.9, 127.9, 127.8, 127.7, 127.6, 127.4, 127.4, 99.0, 80.7, 78.0, 74.7, 74.3, 73.2, 72.5, 72.0, 71.9, 70.6, 68.2, 67.2, 65.0, 50.7, 44.2 (d, J=86.6 Hz), 27.9 (d, J=40.6 Hz). IR (film): v=3482, 3030, 2920, 1698, 1453, 1217, 1092, 1053, 1027, 734, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.53H.sub.57O.sub.9NNa [M+Na].sup.+874.3931, found 874.3916.
[0103] Compound 16*: According to reaction conditions 3.1, the glycosyl donor 13* (472 mg, 0.72 mmol) and the glycosyl receptor 15* (908 mg, 1.06 mmol) reacted to obtain the compound 16* (793 mg, 76%). R.sub.f=0.51, petroleum ether/EtOAc=3:1. [α].sup.25.sub.D=+11.7 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.72-6.78 (m, 50H, arom. H), 5.45 (t, J=2.3 Hz, 1H, 2-H), 5.14 (d, J=11.4 Hz, 2H, Ph—CH.sub.2), 4.89 (s, 1H, 1′-H), 4.88 (s, 1H, 1-H), 4.88-4.30 (m, 14H, Ph—CH.sub.2), 4.25 (d, J=11.0 Hz, 1H, Ph—CH.sub.2), 4.15-4.09 (m, 1H), 3.99 (dt, J=12.5, 5.8 Hz, 2H), 3.94-3.80 (m, 3H), 3.77 (dd, J=10.3, 4.9 Hz, 2H), 3.71 (dd, J=5.8, 2.6 Hz, 2H), 3.67 (dd, J=10.4, 6.5 Hz, 1H, 7-H), 3.50-3.27 (m, 1H, CH.sub.2), 3.27-3.05 (m, 2H, CH.sub.2), 3.05-2.83 (m, 1H, CH.sub.2), 2.07 (s, 3H, CH.sub.3CO), 1.72-1.43 (m, 2H, CH.sub.2). .sup.13C NMR (101 MHz, Chloroform-d) δ169.9, 156.3, 156.3, 138.8, 138.5, 138.4, 138.4, 138.3, 137.8, 128.5, 128.4, 128.3, 128.3, 128.3, 128.2, 128.2, 128.0, 128.0, 127.9, 127.7, 127.7, 127.6, 127.6, 127.6, 127.5, 127.5, 127.4, 127.3, 127.2, 99.9 (C-1), 98.2 (C-1′), 79.9, 78.7, 78.5, 77.6, 76.3, 74.9, 74.5, 74.4, 73.3, 73.1, 72.5, 72.4, 72.3, 72.2, 72.0, 71.7, 71.2, 70.3, 68.6, 67.1, 64.8, 60.4, 50.6, 44.0 (d, J=83.2 Hz), 27.9 (d, J=38.4 Hz), 21.0. IR (film): v=3030, 2922, 1744, 1699, 1454, 1368, 1234, 1095, 1028, 736, 697 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.90H.sub.95O.sub.16NNa [M+Na].sup.+1468.6549, found 1468.6521.
[0104] Compound 17*: According to reaction conditions 3.2, the compound 16* (753 mg, 0.52 mmol) was deacetylated to obtain the compound 17* (657 mg, 90%). R.sub.f=0.32, petroleum ether/EtOAc=3:1. [α].sup.25.sub.D=+13.5 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.50-6.97 (m, 36H, arom. H), 5.14 (d, J=11.3 Hz, 2H), 4.94 (s, 1H, l′-H), 4.93 (s, 1H, 1-H), 4.87 (d, J=10.7 Hz, 1H), 4.77 (d, J=11.0 Hz, 1H), 4.68 (s, 1H), 4.66-4.53 (m, 4H), 4.50-4.34 (m, 7H), 4.10 (d, J=9.6 Hz, 1H), 4.06 (s, 1H), 3.99 (q, J=6.1 Hz, 2H), 3.92 (d, J=9.5 Hz, 1H), 3.85-3.65 (m, 7H), 3.35 (d, J=37.4 Hz, 1H), 3.14 (d, J=34.3 Hz, 2H), 2.93 (d, J=42.3 Hz, 1H), 2.33 (d, J=2.3 Hz, 1H), 1.79-1.44 (m, 2H). .sup.13C NMR (101 MHz, Chloroform-d) δ156.6, 156.0, 138.9, 138.5, 138.5, 138.4, 138.4, 138.3, 137.8, 136.9, 128.5, 128.4, 128.4, 128.3, 128.2, 128.2, 128.2, 127.9, 127.9, 127.8, 127.8, 127.7, 127.7, 127.5, 127.5, 127.4, 127.4, 127.4, 127.3, 127.2, 101.6, 98.3, 80.5, 80.0, 78.9, 76.3, 74.8, 74.6, 74.4, 73.2, 73.1, 72.3, 72.3, 72.2, 71.9, 71.3, 70.2, 68.3, 67.1, 64.8, 50.6, 50.4, 44.4, 43.6, 29.7, 28.1, 27.7. IR (film): v=3030, 2918, 1698, 1453, 1216, 1054, 1027, 734, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.88H.sub.93O.sub.15NNa [M+Na].sup.+1426.6443, found 1426.6480.
[0105] Compound 18*: According to reaction conditions 3.1, the glycosyl donor 11* (258 mg, 0.42 mmol) and the glycosyl receptor 17* (487 mg, 0.42 mmol) reacted to obtain the compound 18* (441 mg, 66%). R.sub.f=0.37, toluene/EtOAc=9:1. [α].sup.25.sub.D=+16.8 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.48-6.99 (m, 60H, arom. H), 5.61-5.54 (m, 1H), 5.38 (d, J=3.3 Hz, 1H), 5.36 (s, 1H), 5.13 (d, J=9.9 Hz, 2H), 5.11 (s, 1H, 1″-H), 4.93 (s, 1H, 1′-H), 4.82 (t, J=10.8 Hz, 3H), 4.75 (s, 1H, 1-H), 4.72-4.57 (m, 7H), 4.53-4.28 (m, 12H), 4.10 (dd, J=10.1, 5.5 Hz, 2H), 3.98 (q, J=6.9, 4.7 Hz, 3H), 3.93-3.63 (m, 12H), 3.54 (dd, J=10.1, 6.6 Hz, 1H), 3.44-3.21 (m, 1H), 3.20-2.96 (m, 2H), 2.98-2.71 (m, 1H), 2.61 (qd, J=10.9, 6.6, 5.4 Hz, 1H), 2.50 (t, J=6.4 Hz, 2H), 2.37 (m, 1H), 2.06 (s, 3H), 1.98 (s, 3H), 1.94 (s, 3H), 1.64-1.47 (m, 2H)..sup.13C NMR (101 MHz, Chloroform-d) δ206.2, 171.8, 169.6, 169.4, 156.2, 156.2, 138.9, 138.8, 138.7, 138.5, 138.5, 138.0, 137.9, 136.8, 128.5, 128.3, 128.3, 128.3, 128.2, 128.2, 128.1, 128.0, 127.9, 127.8, 127.7, 127.7, 127.6, 127.6, 127.6, 127.5, 127.4, 127.4, 127.3, 127.3, 127.1, 101.2, 99.5, 98.4, 79.9, 79.6, 78.9, 74.9, 74.8, 74.7, 74.5, 74.4, 73.5, 73.1, 73.1, 72.9, 72.4, 72.2, 72.1, 71.7, 71.3, 70.2, 70.0, 68.1, 67.1, 65.0, 65.0, 60.4, 50.7, 50.4, 44.5, 43.7, 37.7, 29.7, 28.2, 27.8, 20.8, 20.8, 14.2. IR (film): v=3030, 2919, 1749, 1698, 1453, 1365, 1238, 1216, 1070, 1027, 735, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.118H.sub.131O.sub.25N.sub.2 [M+NH.sub.4].sup.+1975.9035, found 1975.9043.
[0106] Compound 19*: The compound 18* (550 mg, 0.28 mmol) was dissolved in CH.sub.2Cl.sub.2/MeOH (20/1) (3.3 mL), hydrazine acetate (40 mg, 0.42 mmol) was added, and the reaction was performed under stirring at room temperature for 3 h. After it was detected by TLC that the reaction was complete, an appropriate amount of DCM was added for dilution. Then the reaction mixture was washed with saturated NaHCO.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, concentrated, and separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate: 6/1-4/1) to obtain the compound 19* (494 mg, 95%). R.sub.f=0.35, petroleum ether/EtOAc=3:1. [α].sup.25.sub.D=+25.2 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.49-7.02 (m, 60H, arom. H), 5.39 (m, 2H), 5.15 (s, 2H, 1.sup.c-H), 5.10 (s, 1H), 4.97 (s, 1H, 1.sup.a-H), 4.88 (s, 1H, 1.sup.b-H), 4.78 (d, J=11.0 Hz, 2H), 4.74-4.26 (m, 19H), 4.12 (m, 2H), 4.05-3.67 (m, 15H), 3.57 (dd, J=9.9, 4.4 Hz, 1H), 3.48 (dd, J=9.8, 7.4 Hz, 1H), 3.42-3.19 (m, 1H), 3.16-3.01 (m, 2H), 3.01-2.77 (m, 1H), 2.66 (s, 1H), 2.07 (s, 3H, OAc), 1.93 (s, 3H, OAc), 1.69-1.41 (m, 2H, CH.sub.2). .sup.13C NMR (101 MHz, Chloroform-d) δ169.8, 169.6, 156.8, 156.2, 139.1, 139.0, 138.9, 138.7, 138.7, 138.6, 138.6, 138.0, 128.7, 128.6, 128.6, 128.5, 128.4, 128.4, 128.3, 128.2, 128.1, 128.0, 127.9, 127.8, 127.8, 127.6, 127.5, 127.4, 127.4, 127.3, 101.3, 99.3, 98.4, 80.2, 79.7, 79.2, 76.7, 75.5, 75.1, 75.0, 74.8, 74.6, 73.7, 73.6, 73.4, 73.3, 73.3, 73.1, 72.5, 72.4, 72.3, 71.6, 70.9, 70.6, 70.2, 67.3, 65.1, 28.0, 21.1. IR (film): v=3030, 2920, 2360, 1750, 1698, 1453, 1366, 1239, 1218, 1072, 1028, 913, 735, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.113H.sub.125O.sub.23N.sub.2 [M+NH.sub.4].sup.+1877.8668, found 1877.8698.
EXAMPLE 4
Synthesis of repeated disaccharide and trisaccharide
[0107] The synthetic route is shown in
[0108] 4.1 If it is not specified in the disclosure, the conditions for removing the Lev group are as follows: the starting material was dissolved in CH.sub.2Cl.sub.2/MeOH (20/1, 0.1 M), hydrazine acetate (2 eq) was added, and the reaction was performed under stirring at room temperature for 3 hr. After it was detected by TLC that the reaction was complete, an appropriate amount of DCM was added for dissolution. Then the reaction mixture was washed with saturated NaHCO.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, concentrated, and separated and purified by silica gel column chromatography to obtain a Lev-removed receptor.
[0109] Compound 20*: According to reaction conditions 3.1, only the activating reagent TMSOTf (0.12 eq) was added, and the glycosyl donor 8* (674 mg, 0.817 mmol) and the glycosyl receptor 6* (428 mg, 0.68 mmol) reacted to obtain the compound 20* (670 mg, 75%). R.sub.f=0.36, petroleum ether/EtOAc=3:1. [α].sup.25.sub.D=+18.3 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ8.17-6.93 (m, 40H, arom. H), 5.58-5.53 (m, 2H), 5.40 (d, J=1.7 Hz, 2H, 1-H/2-H), 5.21 (d, J=2.0 Hz, 1H, 1-H), 4.94 (d, J=12.1 Hz, 1H), 4.88 (d, J=10.6 Hz, 1H), 4.80 (d, J=12.1 Hz, 1H), 4.73 (d, J=10.7 Hz, 1H), 4.69 (d, J=12.3 Hz, 1H), 4.58 (d, J=11.0 Hz, 1H), 4.55-4.45 (m, 3H), 4.39-4.35 (m, 2H), 4.30 (dd, J=9.5, 3.3 Hz, 1H), 4.27 (d, J=7.2 Hz, 1H), 4.20 (t, J=9.6 Hz, 1H), 4.06-3.94 (m, 3H), 3.78 (dd, J=10.2, 5.0 Hz, 1H), 3.70 (dd, J=10.2, 6.7 Hz, 1H), 3.52 (dd, J=10.5, 7.8 Hz, 1H), 3.29 (dd, J=10.5, 3.6 Hz, 1H), 2.71-2.46 (m, 5H), 2.43-2.27 (m, 2H, CH.sub.2), 2.03 (s, 3H, CH.sub.3CO), 1.25 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ206.1, 171.6, 165.7, 165.2, 139.3, 138.6 (d, J=2.7 Hz), 138.3, 138.0, 137.8, 133.3, 129.9, 129.7, 129.7, 129.4, 128.6, 128.5, 128.3, 128.3, 128.2, 128.2, 127.5, 127.4, 127.4, 127.4, 127.3, 127.3, 127.1, 99.7, 82.0, 79.7, 79.4, 78.5, 75.3, 75.0, 74.2, 74.0, 73.6, 73.5, 73.3, 72.8, 72.6, 72.5, 72.5, 71.8, 70.9, 70.3, 37.8, 29.7, 28.0, 25.5, 14.9. IR (film): v=3030, 2870, 1720, 1452, 1264, 1148, 1093, 1026, 736, 712, 697 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.77H.sub.84O.sub.16SN [M+NH.sub.4].sup.+1310.5505, found 1310.5540.
[0110] Compound 21*: According to the reaction conditions 4.1, the Lev group of the compound 20* (440 mg, 0.34 mmol) was removed to obtain the compound 21* (405 mg, quan.). R.sub.f=0.45, petroleum ether/EtOAc=3:1. [α].sup.25.sub.D=+18.3 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ8.20-6.99 (m, 40H, arom. H), 5.56 (dd, J=3.1, 1.7 Hz, 1H), 5.36 (d, J=1.6 Hz, 1H), 5.34 (dd, J=3.2, 1.8 Hz, 1H), 5.25 (d, J=1.7 Hz, 1H), 4.92 (d, J=12.1 Hz, 1H), 4.85-4.75 (m, 3H), 4.70 (dd, J=11.3, 7.8 Hz, 3H), 4.60 (d, J=11.4 Hz, 1H), 4.49 (d, J=1.2 Hz, 2H), 4.46 (d, J=12.2 Hz, 1H), 4.41-4.36 (m, 2H), 4.32 (dd, J=9.3, 3.1 Hz, 1H), 4.22 (t, J=9.6 Hz, 1H), 4.09 (pd, J=6.6, 6.0, 3.5 Hz, 1H), 4.01 (dq, J=5.5, 2.6, 1.9 Hz, 4H), 3.79 (dd, J=10.2, 5.1 Hz, 1H), 3.70 (dd, J=10.2, 6.7 Hz, 1H), 3.63 (dd, J=10.4, 7.5 Hz, 1H), 3.52 (dd, J=10.3, 4.3 Hz, 1H), 2.72-2.46 (m, 2H, CH.sub.2), 1.91 (d, J=5.0 Hz, 1H, OH), 1.23 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ165.8, 165.4, 139.2, 138.7, 138.6, 138.3, 138.2, 137.7, 133.3, 133.2, 129.8, 129.8, 129.5, 128.6, 128.4, 128.3, 128.3, 128.2, 128.2, 128.0, 127.8, 127.6, 127.5, 127.5, 127.4, 127.4, 127.3, 127.2, 99.4, 82.1, 79.2, 78.6, 77.7, 75.6, 75.4, 75.1, 74.2, 73.8, 73.3, 73.2, 73.0, 72.8, 72.5, 72.5, 71.4, 71.0, 70.4, 25.4, 14.8. IR (film): v=3050, 2926, 1720, 1452, 1265, 1093, 1070, 1026, 825, 736, 711, 698 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.72H.sub.78O.sub.14SN [M+NH.sub.4].sup.+1212.5138, found 1212.5189.
[0111] Compound 22*: According to reaction conditions 3.1, only the activating reagent TMSOTf (0.12 eq) was added, and the glycosyl donor 8* (239 mg, 0.29 mmol) and the glycosyl receptor 21* (288 mg, 0.24 mmol) reacted to obtain the compound 22* (285 mg, 64%). R.sub.f=0.23, petroleum ether/EtOAc=3:1. [α].sup.25.sub.D=+10.2 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ8.24-6.85 (m, 60H, arom. H), 5.57 (s, 1H), 5.56-5.52 (m, 1H), 5.49-5.42 (m, 1H), 5.39-5.36 (m, 1H), 5.37 (d, J=1.8 Hz, 1H, lc-H), 5.33 (d, J=1.9 Hz, 1H, 1.sup.b-H), 4.97 (d, J=1.9 Hz, 1H, 1.sup.a-H), 4.93 (d, J=10.6 Hz, 1H), 4.87 (dd, J=11.6, 5.7 Hz, 2H), 4.78 (d, J=12.1 Hz, 1H), 4.75-4.62 (m, 4H), 4.58-4.50 (m, 2H), 4.49-4.41 (m, 2H), 4.41-4.31 (m, 3H), 4.28 (d, J=12.3 Hz, 1H), 4.17 (t, J=9.8 Hz, 1H), 4.10 (d, J=12.4 Hz, 1H), 3.97 (ddd, J=11.9, 7.3, 4.2 Hz, 3H), 3.87 (d, J=9.7 Hz, 1H), 3.82 (dd, J=8.5, 2.9 Hz, 1H), 3.72 (dd, J=10.3, 4.8 Hz, 1H), 3.65 (dd, J=10.3, 6.9 Hz, 1H), 3.57 (dd, J=10.4, 7.6 Hz, 1H), 3.40-3.30 (m, 2H), 3.03 (dd, J=10.5, 3.0 Hz, 1H), 2.71-2.42 (m, 4H, CH.sub.2), 2.40-2.23 (m, 2H, CH.sub.2), 2.01 (s, 3H, CH.sub.3CO), 1.22 (t, J=7.4 Hz, 3H, CH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ206.1, 171.6, 165.5, 165.5, 165.1, 139.4, 139.2, 138.7, 138.7, 138.3, 137.9, 137.9, 137.7, 133.2, 133.2, 129.9, 129.8, 129.8, 129.7, 129.4, 129.4, 128.5, 128.4, 128.3, 128.2, 128.2, 128.1, 128.1, 128.1, 127.5, 127.5, 127.4, 127.4, 127.4, 127.3, 127.2, 127.2, 127.2, 127.1, 127.0, 127.0, 99.8, 98.7, 82.0, 80.3, 79.2, 78.6, 78.2, 75.4, 75.3, 74.9, 74.2, 74.0, 73.7, 73.6, 73.2, 73.2, 72.9, 72.6, 72.5, 72.4, 72.2, 72.1, 71.6, 71.0, 70.2, 37.8, 29.7, 27.9, 25.4, 14.9. IR (film): v=3030, 2868, 1721, 1452, 1263, 1149, 1092, 1026, 735, 711, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.112H.sub.118O.sub.23SN [M+NH.sub.4].sup.+1876.7810, found 1876.7877.
[0112] The synthetic routes of reducing-end pentasaccharide and octasaccharide are shown in
[0113] Compound 24*: According to reaction conditions 3.1, the glycosyl donor 20* (417 mg, 0.30 mmol, 1.2 equiv) and the glycosyl receptor 23* (452 mg, 0.243 mmol, 1 equiv) reacted to obtain the compound 24* (488 mg, 65%). R.sub.f=0.18, petroleum ether/EtOAc=2:1. [α].sup.25.sub.D=+14.8 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ8.04 (d, J=7.7 Hz, 2H, arom. H), 7.93 (d, J=7.6 Hz, 2H, arom. H), 7.69-6.82 (m, 96H, arom. H), 5.73 (t, J=2.4 Hz, 1H), 5.61 (t, J=2.5 Hz, 1H), 5.49 (d, J=1.8 Hz, 1H, 1-H), 5.45 (dd, J=9.8, 3.2 Hz, 1H), 5.41 (dd, J=9.0, 2.9 Hz, 1H), 5.37 (d, J=2.3 Hz, 1H), 5.22 (s, 1H, 1-H), 5.10 (d, J=4.5 Hz, 2H), 4.94 (s, 1H, 1-H), 4.91 (d, J=2.5 Hz, 1H, 1-H), 4.90-4.81 (m, 3H), 4.80 (d, J=1.4 Hz, 1H, 1-H), 4.79-4.70 (m, 4H), 4.70-4.49 (m, 7H), 4.49-4.08 (m, 23H), 4.04 (d, J=9.8 Hz, 1H), 4.00-3.64 (m, 16H), 3.62-3.50 (m, 2H), 3.45 (tq, J=7.5, 4.2, 3.2 Hz, 3H), 3.39-3.15 (m, 2H), 3.16-2.93 (m, 2H), 2.94-2.70 (m, 1H), 2.61 (dt, J=18.2, 7.3 Hz, 1H), 2.50 (dt, J=18.2, 6.4 Hz, 1H), 2.35 (q, J=6.8 Hz, 2H), 2.08 (s, 3H, OAc), 2.02 (s, 3H, OAc), 1.77 (s, 3H, CH.sub.3CO), 1.61-1.41 (m, 2H, CH.sub.2). .sup.13C NMR (101 MHz, Chloroform-d) δ206.0, 171.6, 170.2, 169.5, 165.5, 165.2, 139.1, 139.1, 139.0, 138.9, 138.8, 138.7, 138.6, 138.6, 138.2, 138.2, 138.1, 138.0, 137.9, 137.8, 136.9, 133.3, 129.9, 129.7, 129.7, 129.5, 128.6, 128.5, 128.4, 128.3, 128.3, 128.3, 128.3, 128.2, 128.2, 128.1, 128.1, 128.1, 128.0, 127.9, 127.8, 127.7, 127.7, 127.7, 127.5, 127.5, 127.4, 127.3, 127.3, 127.3, 127.2, 127.2, 127.1, 127.1, 101.4, 100.3, 99.1, 98.3, 96.0, 80.1, 79.6, 79.3, 79.1, 78.9, 77.5, 76.0, 75.9, 75.3, 75.2, 75.0, 74.8, 74.7, 74.6, 74.2, 73.4, 73.4, 73.3, 73.1, 73.1, 73.0, 73.0, 72.9, 72.9, 72.7, 72.7, 72.4, 72.3, 72.3, 72.3, 72.2, 72.1, 72.1, 71.6, 71.5, 71.4, 70.5, 70.4, 69.8, 67.1, 65.0, 50.6, 43.7, 37.8, 29.6, 28.5, 28.0, 20.6, 20.6, 7.9. IR (film): v=3030, 2917, 1748, 1722, 1453, 1365, 1265, 1240, 1071, 1026, 988, 734, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.188H.sub.195NO.sub.39Na.sub.2 [M+2Na].sup.2+1568.1545, found 1568.1525.
[0114] Compound 25*: According to the reaction conditions 4.1, the Lev group of the compound 24* (393 mg, 0.127 mmol) was removed to obtain the compound 25* (353 mg, 93%). R.sub.f=0.48, petroleum ether/EtOAc=2:1. [α].sup.25.sub.D=+19.2 (c 1.0, CH.sub.3Cl). .sup.1H NMR (600 MHz, Chloroform-d) δ8.03 (d, J=7.7 Hz, 2H, arom. H), 7.92 (d, J=7.7 Hz, 2H, arom. H), 7.69-6.87 (m, 94H, arom. H), 5.77 (s, 1H), 5.53 (s, 1H), 5.47 (dd, J=9.9, 3.1 Hz, 1H), 5.44 (d, J=3.2 Hz, 1H), 5.38 (d, J=3.1 Hz, 1H), 5.31 (s, 1H), 5.13 (d, J=9.9 Hz, 2H), 4.98 (d, J=22.1 Hz, 1H), 4.94-4.85 (m, 5H), 4.84-4.59 (m, 14H), 4.59-4.47 (m, 5H), 4.47-4.10 (m, 24H), 4.06 (d, J=9.8 Hz, 1H), 4.02-3.93 (m, 7H), 3.90 (t, J=9.7 Hz, 1H), 3.87-3.68 (m, 8H), 3.62 (t, J =9.3 Hz, 1H), 3.58-3.47 (m, 4H), 3.44 (dd, J=10.4, 4.2 Hz, 1H), 3.37-3.21 (m, 1H), 3.18-2.96 (m, 3H), 2.98-2.72 (m, 1H), 2.08 (s, 3H, CH.sub.3CO), 1.97 (d, J=5.0 Hz, 1H, 3-OH), 1.76 (s, 3H, CH.sub.3CO), 1.64-1.46 (m, 2H, CH.sub.2). .sup.13C NMR (101 MHz, Chloroform-d) δ170.1, 169.5, 165.9, 165.4, 139.2, 139.1, 139.0, 138.9, 138.8, 138.7, 138.6, 138.2, 138.2, 138.1, 138.0, 137.9, 136.9, 133.3, 133.2, 129.8, 129.8, 129.6, 128.6, 128.5, 128.4, 128.4, 128.3, 128.3, 128.3, 128.1, 128.0, 127.9, 127.9, 127.8, 127.8, 127.7, 127.6, 127.6, 127.5, 127.4, 127.4, 127.3, 127.2, 127.2, 127.2, 127.1, 101.4, 100.0, 99.1, 98.3, 96.0, 80.1, 79.5, 79.3, 78.9, 78.6, 78.4, 77.5, 76.0, 75.7, 75.5, 75.2, 75.0, 74.8, 74.6, 74.1, 73.4, 73.3, 73.2, 73.2, 73.1, 73.0, 73.0, 72.8, 72.4, 72.4, 72.3, 72.2, 72.1, 72.1, 71.6, 71.6, 71.3, 70.8, 70.5, 69.8, 67.1, 65.0, 60.4, 50.7, 31.6, 29.1, 22.6, 20.6, 20.6, 14.1, 14.1. IR (film): v=3030, 2918, 1749, 1722, 1453, 1266, 1071, 1027, 734, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.183H.sub.189NO.sub.37Na.sub.2 [M+2Na].sup.2+1519.1361, found 1519.1348.
[0115] Compound 26*: According to reaction conditions 3.1, the glycosyl donor 22* (228 mg, 0.126 mmol) and the glycosyl receptor 25* (314 mg, 0.105 mmol) reacted to obtain the compound 26* (306 mg, 61%). R.sub.f=0.29, petroleum ether/EtOAc=2:1. [α].sup.25.sub.D=+4.6 (c 0.5, CH.sub.3Cl). .sup.1H NMR (600 MHz, Chloroform-d) δ8.02 (d, J=7.7 Hz, 2H, arom. H), 7.98 (d, J=7.7 Hz, 2H, arom. H), 7.88 (dd, J=12.9, 7.8 Hz, 4H, arom. H), 7.82 (d, J=7.7 Hz, 2H, arom. H), 7.62-6.72 (m, 150H), 5.67 (s, 1H), 5.61 (s, 1H), 5.46 (s, 3H), 5.41 (d, J=9.8 Hz, 1H), 5.37 (s, 1H), 5.33 (s, 2H), 5.09 (d, J=8.0 Hz, 3H), 5.03 (s, 1H), 4.99 (d, J=10.7 Hz, 1H), 4.93 (d, J=21.1 Hz, 1H), 4.85 (s, 3H), 4.77 (s, 1H), 4.68 (ddd, J=16.6, 11.0, 6.0 Hz, 4H), 4.63-4.55 (m, 5H), 4.55-4.42 (m, 8H), 4.34 (tdd, J=28.2, 17.9, 10.5 Hz, 11H), 4.25-4.10 (m, 10H), 4.10-3.92 (m, 9H), 3.92-3.82 (m, 4H), 3.82-3.64 (m, 15H), 3.60-3.22 (m, 9H), 3.17 (d, J=10.1 Hz, 1H), 3.12-3.02 (m, 1H), 2.97 (s, 1H), 2.92-2.79 (m, 3H), 2.75 (d, J=8.8 Hz, 1H), 2.55 (dt, J=18.2, 7.3 Hz, 1H), 2.45 (dt, J=18.2, 6.5 Hz, 1H), 2.30 (tt, J=18.1, 8.8 Hz, 3H, CH.sub.3CO), 2.05 (s, 3H, CH.sub.3CO), 1.98 (s, 3H, CH.sub.3CO), 1.70 (s, 3H, CH.sub.3CO), 1.54 (tt, J=14.6, 6.7 Hz, 1H, CH.sub.2), 1.48-1.38 (m, 1H, CH.sub.2). .sup.13C NMR (101 MHz, Chloroform-d) δ206.0, 171.6, 170.3, 169.6, 165.4, 165.4, 165.2, 165.1, 139.5, 139.5, 139.2, 139.2, 138.9, 138.9, 138.8, 138.8, 138.7, 138.6, 138.6, 138.2, 138.1, 137.9, 137.8, 137.6, 137.6, 136.9, 133.2, 129.9, 129.9, 129.7, 129.5, 129.5, 129.3, 128.6, 128.6, 128.5, 128.4, 128.3, 128.3, 128.2, 128.1, 128.0, 127.9, 127.8, 127.7, 127.6, 127.5, 127.4, 127.3, 127.3, 127.2, 127.2, 127.1, 127.1, 126.9, 126.8, 101.4, 99.9, 99.6, 99.1, 99.0, 98.3, 96.0, 80.6, 80.5, 80.1, 79.6, 79.2, 78.9, 75.9, 75.5, 75.1, 75.0, 74.6, 74.1, 73.9, 73.8, 73.6, 73.6, 73.5, 73.3, 73.1, 72.9, 72.9, 72.8, 72.7, 72.6, 72.6, 72.5, 72.3, 72.2, 72.1, 71.8, 71.7, 71.5, 71.4, 70.5, 70.3, 69.8, 67.1, 65.0, 60.4, 50.6, 37.7, 29.6, 27.9, 21.0, 20.6. IR (film): v=3030, 2922, 2360, 1722, 1452, 1365, 1264, 1092, 1026, 733, 696 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.293H.sub.305N.sub.3O.sub.60 [M+2NH.sub.4].sup.2+2412.5448, found 2412.5464.
[0116] Compound 27*: According to the reaction conditions 4.1, the Lev group of the compound 26* (300 mg, 0.063 mmol) was removed to obtain the compound 27* (264 mg, 90%). R.sub.f=0.46, petroleum ether/EtOAc=2:1. [α].sup.25.sub.D=+25.6 (c 1.0, CH.sub.3Cl). .sup.1H NMR (600 MHz, Chloroform-d) δ8.02 (d, J=7.7 Hz, 2H, arom. H), 7.98 (d, J=7.7 Hz, 2H, arom. H), 7.91 (d, J=7.6 Hz, 2H, arom. H), 7.86 (d, J=7.7 Hz, 2H, arom. H), 7.81 (d, J=7.7 Hz, 2H, arom. H), 7.51 (t, J=7.5 Hz, 2H, arom. H), 7.43 (q, J=7.9 Hz, 3H, arom. H), 7.37-6.79 (m, 145H, arom. H), 5.68 (s, 1H), 5.61 (s, 1H), 5.48-5.45 (m, 2H), 5.44 (s, 1H), 5.41 (dd, J=10.0, 2.9 Hz, 1H), 5.33 (s, 2H), 5.13-5.07 (m, 5H), 5.03 (s, 1H), 4.99 (d, J=10.6 Hz, 1H), 4.95 (s, OH), 4.90 (s, 1H), 4.88-4.80 (m, 5H), 4.80-4.73 (m, 3H), 4.72-4.64 (m, 5H), 4.63-4.55 (m, 5H), 4.55-4.48 (m, 6H), 4.48-4.25 (m, 16H), 4.24-4.11 (m, 10H), 4.11-3.82 (m, 15H), 3.81-3.63 (m, 18H), 3.53 (t, J=9.1 Hz, 1H), 3.47 (t, J=9.0 Hz, 1H), 3.44-3.37 (m, 1H), 3.35 (dd, J=10.5, 3.6 Hz, 1H), 3.28 (dq, J=20.0, 9.8 Hz, 3H), 3.17 (d, J=10.2 Hz, 1H), 3.07 (s, 1H), 3.01-2.93 (m, 2H), 2.87 (d, J=10.1 Hz, 3H), 2.75 (s, 1H), 2.05 (s, 3H, CH.sub.3CO), 1.76 (d, J=5.3 Hz, 1H, 3-OH), 1.69 (s, 3H, CH.sub.3CO), 1.47-1.40 (m, 1H, CH.sub.2). .sup.13C NMR (101 MHz, Chloroform-d) δ170.4, 169.7, 165.7, 165.4, 165.3, 165.3, 165.2, 156.5, 139.5, 139.4, 139.2, 139.1, 138.9, 138.8, 138.7, 138.7, 138.6, 138.5, 138.2, 138.1, 138.0, 137.8, 137.7, 137.6, 137.5, 136.8, 133.3, 129.9, 129.9, 129.8, 129.6, 129.4, 129.3, 128.7, 128.6, 128.5, 128.4, 128.3, 128.3, 128.3, 128.2, 128.2, 128.2, 128.1, 128.1, 128.0, 127.9, 127.9, 127.8, 127.8, 127.7, 127.7, 127.6, 127.4, 127.4, 127.4, 127.3, 127.2, 127.2, 127.1, 127.1, 127.0, 126.9, 126.9, 101.4, 99.5, 99.0, 98.2, 95.9, 80.6, 80.4, 80.2, 80.0, 79.7, 79.6, 79.2, 78.9, 75.7, 75.5, 75.1, 75.0, 74.7, 74.5, 74.1, 73.8, 73.6, 73.6, 73.5, 73.2, 73.1, 72.9, 72.8, 72.8, 72.6, 72.6, 72.5, 72.4, 72.3, 72.2, 72.1, 71.8, 71.7, 71.4, 70.5, 69.7, 67.1, 50.6, 50.4, 44.4, 43.6, 28.2, 27.7, 20.7, 20.7. IR (film): v=3033, 2932, 1726, 1498, 1455, 1267, 1096, 737, 698 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.288H.sub.299N.sub.3O.sub.60 [M+2NH.sub.4].sup.2+2363.5264, found 2363.5398.
EXAMPLE 5
Synthesis of O-antigen tridecasaccharide of Helicobacter pylori serotype O:6
[0117] The synthetic route is shown in
[0118] Compound 30*: According to reaction conditions 3.1, the activating reagents TfOH (0.2 eq) and NIS (1.2 eq) were used, and the glycosyl donor 29* (39 mg, 0.036 mmol) and the glycosyl receptor 28* (22 mg, 0.043 mmol) reacted to obtain the compound 30* (41 mg, 70%). R.sub.f=0.35, Hexane/EtOAc=3:2. [α].sup.25.sub.D=−12.0 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ8.00-7.94 (m, 2H, arom. H), 7.65-7.10 (m, 33H, arom. H), 6.36 (d, J=9.3 Hz, 1H, NH), 5.49 (dd, J=10.0, 7.6 Hz, 1H), 5.11 (dd, J=10.0, 7.9 Hz, 1H), 5.00 (d, J=11.6 Hz, 1H, Ph—CH.sub.2), 4.84 (d, J=11.4 Hz, 1H, Ph—CH.sub.2), 4.67 (d, J=7.8 Hz, 1H, 1-H), 4.64-4.56 (m, 3H), 4.48-4.39 (m, 6H), 4.37 (d, J=11.7 Hz, 1H, Ph—CH.sub.2), 4.31 (d, J=7.9 Hz, 1H, 1-H), 4.03 (dd, J=10.2, 3.1 Hz, 1H), 4.01-3.88 (m, 5H), 3.81 (dd, J=11.1, 3.5 Hz, 1H), 3.73 (dd, J=11.1, 1.8 Hz, 1H), 3.65 (t, J=6.1 Hz, 1H), 3.61-3.48 (m, 5H), 3.38 (dd, J=8.3, 5.1 Hz, 1H), 3.33 (dd, J=10.1, 2.8 Hz, 1H), 2.77 (ddd, J=18.3, 8.0, 5.7 Hz, 1H, CH.sub.2), 2.57 (dt, J=18.3, 6.0 Hz, 1H, CH.sub.2), 2.52-2.41 (m, 2H, CH.sub.2), 2.40-2.26 (m, 4H, CH.sub.2), 2.15 (s, 3H, CH.sub.3CO), 1.88 (s, 3H, CH.sub.3CO), 0.69 (s, 9H, CH.sub.3), 0.01 (s, 3H, SiCH.sub.3), −0.07 (s, 3H, SiCH.sub.3). .sup.13C NMR (101 MHz, Chloroform-d) δ206.5, 206.4, 172.5, 171.1, 164.7, 162.0, 138.6, 138.5, 138.1, 138.0, 137.9, 137.7, 133.2, 130.0, 129.7, 128.9, 128.6, 128.5, 128.5, 128.4, 128.4, 128.2, 128.1, 128.1, 128.0, 128.0, 127.9, 127.8, 127.7, 127.5, 127.5, 127.4, 101.2, 100.5, 96.7, 91.9, 80.2, 78.9, 75.9, 75.0, 74.8, 74.6, 74.2, 74.1, 74.0, 73.5, 73.1, 72.3, 72.2, 71.9, 71.7, 69.2, 67.8, 67.7, 55.9, 37.8, 37.7, 30.0, 29.6, 27.9, 27.8, 25.4, 17.7, −4.1, −5.4. IR (film): v=1720, 1071, 838, 700 cm .sup.−1.
[0119] Compound 31*: The compound 30* (40 mg, 0.025 mmol) was dissolved in THF (1 mL), and then acetic acid (14 μL, 0.25 mmol) was added and stirred. TBAF/THF (1 M, 0.25 mL) was added at 0° C., and then the reaction was performed under stirring at room temperature for 4 h. After it was detected by TLC that the reaction was complete, an appropriate amount of DCM was added for dissolution. Then the reaction solution was washed with saturated NaHCO.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, filtered, concentrated, and separated and purified by column chromatography to obtain corresponding hemiacetal.
[0120] The obtained hemiacetal was dissolved in CH.sub.2Cl.sub.2 (2 mL), CCl.sub.3CN (18 μL, 0.125 mmol) and DBU (11 μL, 0.075 mmol) were added at 0° C., and the reaction was performed under stirring at room temperature for 1.5 min. After it was detected by TLC that the reaction was complete, the reaction solution was concentrated at 30° C., and then separated and purified by silica gel column chromatography (n-hexane/ethyl acetate: 2/1.fwdarw.1/1) to obtain the compound 31* (37 mg, 87%). R.sub.f=0.44, Hexane/EtOAc=1:1. [α].sup.25.sub.D=7.3 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ8.03 (d, J=7.8 Hz, 2H, arom. H), 7.72-6.96 (m, 38H, arom. H), 6.57 (d, J=7.7 Hz, 2H), 6.41 (d, J=9.3 Hz, 1H), 5.91 (s, 1H), 5.77 (t, J=9.1 Hz, 1H), 5.21-5.01 (m, 2H), 5.00-4.77 (m, 2H), 4.77-4.56 (m, 4H), 4.55-4.26 (m, 9H), 4.26-3.88 (m, 5H), 3.90-3.69 (m, 2H), 3.70-3.49 (m, 6H), 3.37 (ddd, J=22.6, 9.1, 4.0 Hz, 2H), 2.79 (ddd, J=18.3, 8.3, 5.4 Hz, 1H, CH.sub.2), 2.67-2.26 (m, 7H, CH.sub.2), 2.17 (s, 3H, CH.sub.3CO), 1.90 (s, 3H, CH.sub.3CO). .sup.13C NMR (101 MHz, Chloroform-d) δ206.5, 206.3, 172.6, 171.1, 164.6, 162.0, 143.2, 138.5, 138.3, 137.9, 137.8, 137.7, 137.6, 133.7, 129.8, 129.2, 128.9, 128.8, 128.6, 128.5, 128.4, 128.2, 128.0, 128.0, 127.9, 127.8, 127.8, 127.6, 127.6, 127.4, 124.2, 119.2, 101.4, 100.5, 95.4, 91.9, 80.2, 78.6, 75.4, 75.1, 75.0, 74.6, 74.0, 73.5, 73.1, 72.3, 72.0, 71.9, 71.8, 71.6, 67.7, 55.9, 37.7, 37.7, 30.0, 29.6, 27.9, 27.8. IR (film): v=2873, 1719, 1456, 1266 1210, 1164, 1096, 820, 737, 698 cm .sup.−1.
[0121] Compound 32*: According to reaction conditions 3.1, only the activating reagent TMSOTf (0.15 eq) was added, and the glycosyl donor 31* (37 mg, 0.0223 mmol, 1.5 eq) and the glycosyl receptor 27* (70 mg, 0.0149 mmol, 1 eq) reacted to obtain the compound 32* (59 mg, 65%). R.sub.f=0.26, Hexane/EtOAc=3:2. [α].sup.25.sub.D=+11.8 (c 1.0, CH.sub.3Cl). .sup.1H NMR (700 MHz, Chloroform-d) δ8.04 (d, J=7.6 Hz, 2H, arom. H), 7.98 (d, J=7.6 Hz, 2H, arom. H), 7.93 (d, J=7.7 Hz, 2H, arom. H), 7.85 (d, J=7.6 Hz, 2H, arom. H), 7.62-6.56 (m, 187H, arom. H), 6.06 (d, J=9.2 Hz, 1H, NH), 5.69 (s, 1H), 5.62 (d, J=3.1 Hz, 1H), 5.48 (s, 1H), 5.47 (s, 1H), 5.43 (d, J=12.4 Hz, 2H), 5.39-5.32 (m, 3H), 5.18-5.09 (m, 4H), 5.09-5.04 (m, 2H), 5.00 (d, J=10.6 Hz, 1H), 4.97-4.92 (m, 2H), 4.91-4.57 (m, 27H), 4.56-4.29 (m, 33H), 4.27-4.08 (m, 17H), 4.05-3.85 (m, 18H), 3.84-3.65 (m, 15H), 3.63 (d, J=8.8 Hz, 1H), 3.60-3.47 (m, 7H), 3.46-3.33 (m, 7H), 3.30 (t, J=9.5 Hz, 1H), 3.25 (t, J=9.6 Hz, 1H), 3.18 (d, J=9.8 Hz, 1H), 3.10 (dd, J=8.9, 5.0 Hz, 2H), 2.98 (q, J=9.3, 6.7 Hz, 2H), 2.87 (d, J=9.7 Hz, 1H), 2.84-2.75 (m, 1H), 2.73 (d, J=9.9 Hz, 1H), 2.65-2.57 (m, 2H), 2.50 (ddd, J=14.2, 8.2, 5.4 Hz, 1H), 2.44 (dd, J=16.4, 7.5 Hz, 1H), 2.41-2.26 (m, 4H, CH.sub.2), 2.18 (s, 3H, CH.sub.3CO), 2.07 (s, 3H, CH.sub.3CO), 1.87 (s, 3H, CH.sub.3CO), 1.72 (s, 3H, CH.sub.3CO), 1.50-1.41 (m, 1H). .sup.13C NMR (176 MHz, Chloroform-d) δ206.2, 206.1, 172.4, 171.1, 170.3, 169.6, 165.3, 165.2, 165.0, 164.0, 161.8, 139.6, 139.5, 139.5, 139.2, 139.1, 139.0, 138.9, 138.9, 138.8, 138.8, 138.7, 138.6, 138.6, 138.5, 138.5, 138.1, 138.1, 138.0, 138.0, 137.9, 137.9, 137.8, 137.7, 137.4, 132.5, 129.9, 129.9, 129.6, 129.4, 129.3, 129.2, 129.1, 129.0, 128.8, 128.6, 128.6, 128.5, 128.5, 128.4, 128.3, 128.3, 128.2, 128.2, 128.1, 128.1, 128.1, 128.0, 128.0, 128.0, 127.9, 127.9, 127.8, 127.8, 127.8, 127.7, 127.7, 127.6, 127.6, 127.5, 127.5, 127.4, 127.4, 127.3, 127.3, 127.3, 127.2, 127.1, 127.1, 127.1, 127.0, 127.0, 127.0, 126.9, 126.9, 126.8, 126.7, 101.3, 101.1, 100.5, 99.5, 99.0, 98.8, 91.7, 80.8, 80.3, 80.0, 79.2, 78.5, 75.9, 75.8, 75.1, 75.0, 75.0, 74.7, 74.5, 74.5, 74.3, 74.0, 73.9, 73.7, 73.5, 73.5, 73.2, 73.1, 73.1, 72.9, 72.8, 72.7, 72.6, 72.4, 72.4, 72.3, 72.2, 72.2, 72.1, 72.0, 71.8, 71.7, 71.3, 70.5, 69.7, 69.0, 67.7, 67.5, 67.0, 55.7, 37.7, 29.9, 29.5, 27.8, 20.6, 20.6. IR (film): v=2928, 1724, 1455, 1267, 1097, 737, 698 cm.sup.−1.
[0122] Compound 33*: The compound 32* (20 mg, 0.0032 mmol) was dissolved in pyridine (0.5 mL), hydrazine acetate (2 mg, 0.016 mmol) was added, and the reaction was performed under stirring at room temperature for 3 h. After it was detected by TLC that the reaction was complete, the pyridine was removed by concentration, and an appropriate amount of DCM was added for dilution. Then the reaction mixture was sequentially washed with 1 M HCl, a saturated NaHCO.sub.3 and a saline solution, dried with anhydrous Na.sub.2SO.sub.4, concentrated, and separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate: 2/1.fwdarw.3/2) to obtain the compound 33* (17 mg, 89%). R.sub.f=0.34, Hexane/EtOAc=3:2. [α].sup.25.sub.D=+13.4 (c 1.0, CH.sub.3Cl). .sup.1H NMR (700 MHz, Chloroform-d) δ8.04 (d, J=7.6 Hz, 2H, arom. H), 7.98 (d, J=7.7 Hz, 2H, arom. H), 7.93 (d, J=7.7 Hz, 2H, arom. H), 7.86 (d, J=7.5 Hz, 2H, arom. H), 7.53-6.55 (m, 187H, arom. H), 6.04 (d, J=8.6 Hz, 1H, NH), 5.69 (s, 1H), 5.63 (s, 1H), 5.48 (s, 1H), 5.47 (s, 2H), 5.42 (q, J=10.5 Hz, 3H), 5.34 (d, J=7.8 Hz, 2H), 5.12 (d, J=10.4 Hz, 3H), 5.08 (s, 2H), 5.01 (d, J=10.7 Hz, 1H), 4.98-4.90 (m, 3H), 4.90-4.45 (m, 40H), 4.37 (ddt, J=45.4, 24.8, 10.6 Hz, 13H), 4.28-4.15 (m, 11H), 4.15-4.07 (m, 6H), 4.01 (dt, J=20.8, 9.3 Hz, 10H), 3.94-3.84 (m, 6H), 3.83-3.65 (m, 16H), 3.62 (d, J=9.1 Hz, 2H), 3.52 (tq, J=28.2, 9.6, 8.4 Hz, 7H), 3.37 (dtt, J=50.0, 18.4, 10.2 Hz, 7H), 3.25 (t, J=9.7 Hz, 1H), 3.19 (t, J=13.6 Hz, 1H), 3.10 (d, J=6.6 Hz, 2H), 2.99 (q, J=8.3, 6.7 Hz, 2H), 2.88 (t, J=15.7 Hz, 1H), 2.81-2.75 (m, 1H), 2.72 (d, J=10.0 Hz, 1H), 2.62 (d, J=10.3 Hz, 1H), 2.07 (s, 3H, CH.sub.3CO), 1.72 (s, 3H, CH.sub.3CO), 1.50-1.41 (m, 1H). .sup.13C NMR (176 MHz, Chloroform-d) δ170.3, 169.6, 165.3, 165.3, 165.2, 165.2, 165.1, 164.1, 161.8, 139.6, 139.5, 139.5, 139.2, 139.1, 139.0, 139.0, 138.8, 138.8, 138.7, 138.6, 138.6, 138.5, 138.4, 138.1, 138.1, 138.1, 137.9, 137.9, 137.8, 137.8, 137.8, 137.7, 137.4, 137.4, 137.3, 133.2, 133.1, 132.8, 132.6, 129.9, 129.9, 129.6, 129.4, 129.3, 129.2, 129.2, 129.1, 129.0, 128.7, 128.6, 128.6, 128.5, 128.5, 128.3, 128.3, 128.3, 128.2, 128.2, 128.2, 128.2, 128.1, 128.1, 128.0, 128.0, 128.0, 127.9, 127.9, 127.9, 127.8, 127.8, 127.7, 127.7, 127.6, 127.6, 127.4, 127.4, 127.3, 127.3, 127.2, 127.1, 127.1, 127.1, 127.0, 127.0, 127.0, 126.9, 126.9, 126.8, 126.7, 104.2, 101.3, 100.3, 99.5, 99.0, 98.8, 97.7, 95.9, 92.1, 82.7, 81.9, 81.2, 81.1, 80.8, 80.5, 80.0, 79.6, 79.2, 78.9, 77.9, 75.8, 75.5, 75.1, 75.0, 74.7, 74.5, 74.5, 74.4, 74.0, 74.0, 73.9, 73.8, 73.7, 73.6, 73.4, 73.2, 73.2, 73.1, 72.9, 72.8, 72.8, 72.6, 72.5, 72.4, 72.3, 72.3, 72.2, 72.1, 71.9, 71.8, 71.7, 71.4, 71.3, 71.0, 70.5, 69.7, 69.0, 68.9, 68.4, 67.5, 67.1, 57.3, 50.6, 45.3, 29.7, 20.6, 20.6. IR (film): v=3432, 3066, 3033, 2926, 2869, 1725, 1603, 1498, 1455, 1367, 1267, 1218, 1096, 1028, 912, 820, 736, 698 cm.sup.−1.
[0123] Compound 35*: The glycosyl donor 34* (7 mg, 0.0107 mmol) and the glycosyl receptor 33* (15 mg, 0.00268 mmol, 1 eq) were mixed, dissolved in toluene and steamed twice. Dry DCM/Et.sub.2O (v/v, 1:1) (0.2 mL) was added, and activated 3 Å or 4 Å A molecular sieves were added. 10 eq of thiophene was added, the reaction temperature was lowered to −40° C., and the reaction mixture was stirred for 15 min. Then the activating reagent TMSOTf (0.4 eq) was added, and the reaction was performed under stirring at −40° C. for 3 h. After it was detected by TLC that the reaction was complete, an appropriate amount of Et.sub.3N was added to terminate the reaction. The reaction solution was filtered, diluted with DCM, washed with saturated NaHCO.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, concentrated, separated and purified by silica gel column chromatography (n-hexane/ethyl acetate: 2/1.fwdarw.3/2) to obtain the compound 35* (9 mg, 52%). R.sub.f=0.48, Hexane/EtOAc=3:2. [α].sup.25.sub.D=−1.5 (c 1.0, CH.sub.3Cl). .sup.1H NMR (700 MHz, Chloroform-d) δ8.02 (d, J=7.7 Hz, 2H, arom. H), 7.96 (d, J=7.6 Hz, 2H, arom. H), 7.90 (d, J=7.5 Hz, 2H, arom. H), 7.83 (d, J=7.5 Hz, 2H, arom. H), 7.60-6.52 (m, 217H, arom. H), 6.35 (d, J=7.6 Hz, 1H, NH), 5.68-5.66 (m, 1H), 5.65 (d, J=4.0 Hz, 1H), 5.62-5.57 (m, 1H), 5.46 (s, 1H), 5.45-5.39 (m, 4H), 5.35-5.30 (m, 1H), 5.32 (s, 2H), 5.12-5.08 (m, 3H), 5.05 (s, 1H), 5.01-4.92 (m, 5H), 4.89 (d, J=4.2 Hz, 1H), 4.87-4.81 (m, 5H), 4.80-4.74 (m, 7H), 4.67 (ddd, J=13.6, 11.0, 6.7 Hz, 5H), 4.62-4.42 (m, 23H), 4.41-4.25 (m, 16H), 4.25-4.04 (m, 22H), 4.03-3.95 (m, 9H), 3.95-3.88 (m, 3H), 3.87-3.56 (m, 23H), 3.55-3.45 (m, 5H), 3.44-3.30 (m, 4H), 3.30-3.25 (m, 2H), 3.22 (td, J=9.8, 5.1 Hz, 2H), 3.16 (d, J=10.2 Hz, 1H), 3.12 (d, J=2.5 Hz, 1H), 3.03 (tq, J=32.8, 11.5, 9.2 Hz, 3H), 2.84 (d, J=9.6 Hz, 1H), 2.79-2.73 (m, 1H), 2.73-2.66 (m, 1H), 2.62 (d, J=9.9 Hz, 1H), 2.05 (s, 3H, CH.sub.3CO), 1.70 (s, 3H, CH.sub.3CO), 1.45 (ddd, J=15.8, 7.5, 4.0 Hz, 1H), 1.12 (d, J=6.3 Hz, 3H, CH.sub.3-Fucose), 1.02 (d, J=6.3 Hz, 3H, CH.sub.3-Fucose). .sup.13C NMR (176 MHz, Chloroform-d) δ170.4, 169.7, 165.5, 165.5, 165.3, 165.2, 164.6, 161.2, 156.4, 156.4, 139.7, 139.7, 139.6, 139.4, 139.2, 139.1, 139.1, 139.0, 139.0, 138.9, 138.8, 138.8, 138.7, 138.6, 138.3, 138.3, 138.2, 138.1, 138.0, 138.0, 137.9, 137.9, 137.8, 137.6, 137.5, 133.4, 133.2, 132.8, 130.1, 130.0, 129.8, 129.6, 129.5, 129.4, 129.3, 129.1, 129.0, 128.8, 128.7, 128.7, 128.6, 128.6, 128.6, 128.5, 128.5, 128.5, 128.5, 128.4, 128.3, 128.3, 128.3, 128.3, 128.2, 128.2, 128.1, 128.1, 128.0, 128.0, 128.0, 127.9, 127.9, 127.9, 127.9, 127.8, 127.8, 127.7, 127.7, 127.6, 127.6, 127.5, 127.5, 127.4, 127.4, 127.4, 127.3, 127.3, 127.2, 127.2, 127.2, 127.1, 127.1, 127.1, 127.0, 126.9, 126.8, 126.2, 101.5, 100.3, 99.6, 99.5, 99.1, 99.0, 98.4, 98.1, 97.7, 97.5, 96.0, 91.8, 84.0, 81.3, 81.2, 81.0, 80.6, 80.1, 79.8, 79.7, 79.4, 79.3, 79.2, 79.1, 78.6, 78.4, 78.3, 76.7, 76.3, 75.9, 75.8, 75.7, 75.6, 75.4, 75.2, 75.1, 75.0, 75.0, 74.9, 74.8, 74.7, 74.6, 74.3, 74.2, 74.0, 73.9, 73.9, 73.8, 73.7, 73.7, 73.5, 73.5, 73.3, 73.3, 73.2, 73.2, 73.1, 73.0, 73.0, 72.9, 72.8, 72.7, 72.6, 72.6, 72.5, 72.4, 72.4, 72.3, 72.3, 72.2, 72.2, 72.1, 72.0, 71.9, 71.8, 71.6, 71.5, 71.2, 70.6, 70.2, 69.8, 68.8, 68.1, 67.8, 67.6, 67.2, 66.9, 66.9, 66.6, 66.3, 65.0, 50.8, 50.5, 44.5, 43.8, 28.3, 27.8, 20.8, 20.8, 16.5, 16.3. IR (film): v=1725, 1266, 1096, 734, 698 cm .sup.−1.
[0124] Compound 36*: The compound 35* (9 mg, 0.0014 mmol, 52%) was dissolved in AcOH (1 mL), newly activated zinc powder (100 mg) was added, and the reaction was performed under stirring at room temperature for 12 h. After it was detected by TLC that the raw materials disappeared, the solution was filtered and concentrated under reduced pressure. Then the solution was diluted with an appropriate amount of DCM, washed with saturated NaHCO.sub.3, and dried with anhydrous Na.sub.2SO.sub.4. The solution was filtered, concentrated under reduced pressure, and dried in vacuum to obtain the NAc intermediate. The intermediate was dissolved in THF/MeOH (1:1, 1 mL), MeONa (30 mg) was added, and the solution was stirred at room temperature for 15 min. Then NaOH (aq, 1 M, 100 μL) was added, and the reaction was performed under stirring at room temperature for 12 h. After it was detected by TLC that the reaction was complete, the reaction solution was neutralized with Amerlite IR 120 (H.sup.+) resin to reach a pH of 7. The solution was filtered, concentrated under reduced pressure, and separated and purified by silica gel column chromatography (dichloromethane/methanol: 50/1) to obtain an intermediate. The deacylated compound was dissolved in MeOH/THF/H.sub.2O/AcOH (10:5:4:1, 12 mL), 10% Pd/C (50 mg) was added, and the reaction mixture was stirred in hydrogen (1 bar) for 24 h. After time-of-flight mass spectrometry detected that the reaction was complete, the solution was filtered and concentrated, dried in vacuum, and then separated and purified on a Sephadex LH.sub.2O gel column to obtain the compound 36*. .sup.1H NMR (700 MHz, Deuterium Oxide) δ5.20 (d, J=2.8 Hz, 1H, 1-H), 5.13 (s, 1H, 1-H), 5.10 (s, 1H, 1-H), 5.09 (s, 1H, 1-H), 5.06-5.01 (m, 5H, 1-H), 4.95 (s, 1H, 1-H), 4.82-4.78 (m, 1H), 4.65 (d, J=8.6 Hz, 1H, 1-H), 4.44 (dd, J=7.8, 4.1 Hz, 2H), 4.21-4.14 (m, 6H), 4.12 (s, 1H), 4.06 (s, 1H), 3.99 (t, J=9.7 Hz, 12H), 3.95-3.86 (m, 10H), 3.86-3.68 (m, 44H), 3.64 (dt, J=21.4, 9.9 Hz, 15H), 3.60-3.50 (m, 6H), 3.41-3.36 (m, 1H), 3.11-3.01 (m, 2H, CH.sub.2), 1.95 (s, 3H, CH.sub.3CO), 1.94-1.89 (m, 2H, CH.sub.2), 1.19 (d, J=6.5 Hz, 3H, Fucose-CH.sub.3), 1.18-1.13 (d, J=6.5 Hz, 3H, Fucose-CH.sub.3). .sup.13C NMR (176 MHz, Deuterium Oxide) δ158.8, 102.1, 102.0, 101.9, 101.7, 101.2, 100.5, 100.2, 99.4, 98.5, 98.1, 82.1, 79.7, 79.0, 78.6, 78.6, 78.2, 77.9, 77.8, 76.3, 75.4, 74.8, 74.8, 74.0, 73.9, 73.8, 73.7, 73.5, 73.2, 73.1, 72.1, 71.9, 71.7, 71.5, 71.4, 71.4, 71.3, 70.6, 70.4, 70.1, 69.9, 69.7, 69.5, 69.4, 69.1, 68.7, 68.4, 68.3, 67.7, 67.7, 67.5, 67.5, 66.9, 66.8, 66.8, 66.7, 66.0, 65.7, 64.9, 62.1, 62.0, 61.7, 61.6, 61.5, 61.5, 61.0, 59.8, 37.5, 26.6, 22.3, 15.4 (d, J=3.2 Hz).
EXAMPLE 6
Different synthesis strategies of tridecasaccharide
[0125] In the course of the experiment, the inventors also tried other different [5+8] and [4+9] synthesis strategies, using a tetrasaccharide (
(1) Synthesis of glycosylation donors tetrasaccharide and pentasaccharide
[0126] The synthetic route is shown in
[0127] The method uses the monosaccharide building block 37* as the receptor to react with the glycosylation donor 38* under the action of an activating reagent TMSOTf to obtain disaccharide 39*. Then the acyl groups Bz and Lev were removed using CH.sub.3ONa to obtain the disaccharide receptor 40* to react with 4 equivalents of the glycosyl donor 34* to obtain the fully protected Le tetrasaccharide. To further obtain a pentasaccharide donor, the compound 28* was used as a glycosylation receptor to react with the donor Ley tetrasaccharide to obtain pentasaccharide 42*. Since the amino protecting group Phth on the donor inactivated the activity of the donor, the reaction yield was only 21%. Then, TBS at the reducing end was removed by TBAF, and then the pentasaccharide 42* was converted into an NPh-based trifluoroacetimidate donor 43* for the subsequent glycosylation reaction.
[0128] To optimize the synthesis of the pentasaccharide, we decided to replace the amino protecting group Phth with Troc to increase the activity of the tetrasaccharide donor. First, the phthaloyl group was removed by ethylenediamine under a heating condition, and then the amino group was protected by Troc to obtain the compound 44*. Similarly, under the action of TfOH and NIS, the compound 44* reacted with the receptor 28* to obtain the compound 45*. Because part of the amino protecting group Troc was removed when TBS was removed by TBAF, we chose HF to remove TBS. Although we successfully removed TBS, we got the by-products with exposed amino groups. After separation and purification, the mixture was transformed into the NPh-based trifluoroacetimidate donor 43* for subsequent glycosylation.
[0129] The experimental procedure is as follows:
[0130] Compound 39*: The compounds 37* (270 mg, 0.50 mmol) and 38* (435 mg, 0.60 mmol) were dissolved in anhydrous CH.sub.2Cl.sub.2 (10 mL), and freshly activated 4 Å molecular sieves were added. The temperature was lowered to −10° C., and then TMSOTf (11 μL, 0.06 mmol) was added dropwise under argon protection. The reaction was performed at −10° C. for 3 h. After it was detected by TLC that the reaction was complete, an appropriate amount of Et.sub.3N was added to quench the reaction. After being filtered and concentrated, the reaction solution was purified by silica gel column chromatography (petroleum ether/ethyl acetate: 20/1.fwdarw.8/1) to obtain the disaccharide compound 39* (330 mg, 63%).
[0131] Compound 40*: The compound 39* (214 mg, 0.20 mmol) was dissolved in THF/MeOH (1/1, v/v, 2 mL), and sodium methoxide (20 mg, 0.40 mmol) was added at room temperature. The reaction was performed overnight at room temperature. After it was detected by TLC that the raw materials reacted completely, methanol was added for dilution. The excess sodium methoxide was neutralized with the resin, the pH was adjusted to 0, and then the solution was filtered, concentrated, and dried in vacuum. The above crude compound was dissolved in pyridine (20 mL), 10% AcOH (2 mL) was added, and the mixture was heated to reflux for 16 h. After it was detected by TLC that the reaction was complete, the reaction solution was concentrated, dried, and purified by silica gel column chromatography (petroleum ether/ethyl acetate: 5/1.fwdarw.2/1) to obtain the compound 40* (150 mg, 86%).
[0132] Compound 41*: The compound 34* (412 mg, 0.68 mmol, 4 eq) and compound 40* (150 mg, 0.17 mmol, 1 eq) were dissolved in anhydrous CH.sub.2Cl.sub.2/Et.sub.2O (1/1, v/v, 3 mL), and 4 Å molecular sieves were added. The reaction temperature was lowered to −40° C., and then TMSOTf (12 μL) was added dropwise under the protection of argon. The reaction was performed for 2.5 h at −40° C. After it was detected by TLC that the reaction was complete, an appropriate amount of Et.sub.3N was added to quench the reaction. The solution was filtered, concentrated, and purified by silica gel column chromatography (petroleum ether/ethyl acetate: 4/1) to obtain the compound 41* (215 mg, 74%). R.sub.f=0.40, Petroleum ether/EtOAc=2:1. [α].sup.25.sub.D=−51.2 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.91-7.65 (m, 4H, arom. H), 7.48-6.81 (m, 50H, arom. H), 5.73 (d, J=3.8 Hz, 1H, 1-H), 5.13 (d, J=10.5 Hz, 1H, 1-H), 5.00 (d, J=11.3 Hz, 1H, Ph—CH.sub.2), 4.84-4.71 (m, 6H, Ph—CH.sub.2), 4.68 (d, J=4.2 Hz, 1H, 1-H), 4.62 (d, J=8.3 Hz, 1H, 1-H), 4.60-4.42 (m, 9H), 4.42-4.33 (m, 2H), 4.29 (d, J=11.6 Hz, 2H), 4.23 (t, J=9.5 Hz, 1H), 4.16-4.07 (m, 2H), 4.02 (d, J=2.9 Hz, 1H), 3.92 (dd, J=11.6, 3.6 Hz, 1H), 3.89-3.82 (m, 2H), 3.81-3.65 (m, 6H), 3.62 (dd, J=9.6, 3.0 Hz, 1H), 3.41 (ddd, J=21.0, 9.6, 4.0 Hz, 2H), 3.11 (d, J=2.4 Hz, 1H), 2.83-2.63 (m, 2H, SCH.sub.2), 1.43 (d, J=6.5 Hz, 3H, CH.sub.3-Fucose,), 1.22 (t, J=7.4 Hz, 3H, CH.sub.3), 1.14 (d, J=6.4 Hz, 3H, CH.sub.3-Fucose). .sup.13C NMR (101 MHz, Chloroform-d) δ168.4, 167.1, 139.2, 138.8, 138.8, 138.7, 138.5, 138.4, 138.1, 138.0, 137.7, 134.3, 134.1, 131.8, 131.6, 128.7, 128.6, 128.4, 128.4, 128.3, 128.2, 128.1, 128.1, 128.0, 127.8, 127.8, 127.6, 127.5, 127.5, 127.4, 127.4, 127.3, 127.3, 127.2, 127.0, 126.9, 126.1, 123.8, 123.5, 100.0, 98.1, 97.9, 83.9, 81.5, 80.6, 79.7, 79.1, 78.3, 78.1, 75.6, 75.4, 74.9, 74.7, 73.9, 73.6, 73.3, 73.2, 73.1, 72.9, 72.8, 72.5, 72.4, 71.7, 70.9, 68.2, 67.7, 67.0, 66.8, 55.6, 23.5, 16.4, 16.2, 14.9. IR (film): v=3032, 2870, 2351, 1778, 1716, 1498, 1455, 1385, 1365, 1208, 1095, 1044, 1028, 912, 815, 735, 697 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.104H.sub.109NO.sub.19SNa [M+Na].sup.+1730.7207, found 1730.7238.
[0133] Compound 42*: The donor tetrasaccharide 41* (35 mg, 0.019 mmol) and the receptor 28* (21 mg, 0.038 mmol) were dissolved in anhydrous CH.sub.2Cl.sub.2 (1 mL), and 4 Å molecular sieves and NIS (5 mg, 0.023 mmol) were added. The reaction temperature was lowered to −20° C., and then TfOH (0.34 μL, 0.0038 mmol) was added dropwise under argon protection. The reaction was performed at −20° C. for 3 h. After it was detected by TLC that the reaction was complete, an appropriate amount of Et.sub.3N was added to quench the reaction. The solution was filtered, washed with a 10% Na.sub.2S.sub.2O.sub.3 solution until the solution was colorless, then sequentially washed with saturated NaHCO.sub.3 and a saturated saline solution, and dried with anhydrous Na.sub.2SO.sub.4. The solution was concentrated and purified by silica gel column chromatography (n-hexane/ethyl acetate: 4/1) to obtain the compound 42* (5.5 mg, 13%).
[0134] Compound 43*: The compound 42* (11 mg, 0.005 mmol) was dissolved in THF (0.5 mL), and then acetic acid (3 μL, 0.05 mmol) was added and stirred. TBAF/THF (1 M, 0.05 mL) was added at 0° C., and then the reaction was performed at room temperature for 4 h. After it was detected by TLC that the reaction was complete, an appropriate amount of DCM was added for dissolution. Then the reaction solution was washed with saturated NaHCO.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, filtered, concentrated, and separated and purified by column chromatography to obtain corresponding hemiacetal.
[0135] The obtained hemiacetal was dissolved in CH.sub.2Cl.sub.2 (0.5 mL), CCl.sub.3CN (4 μL, 0.025 mmol) and DBU (3 μL, 0.019 mmol) were added at 0° C., and the reaction was performed under stirring at room temperature for 1.5 h. After it was detected by TLC that the reaction was complete, the reaction solution was concentrated at 30° C., and then separated and purified by silica gel column chromatography (n-hexane/ethyl acetate: 5/1.fwdarw.3/1) to obtain the compound 43* (8 mg, 71%).
[0136] Compound 44*: The compound 41* (164 mg, 0.096 mmol) was dissolved in n-BuOH (5 mL), ethylenediamine (5 mL) was added, and the reaction was performed at 95° C. for 6 h. The reaction mixture was concentrated under reduced pressure, and azeotropic evaporation was performed twice using toluene to obtain the crude compound. The crude product was dissolved in pyridine (2 mL), then TrocCl (33 μL, 0.24 mmol) was added, and the reaction was performed under stirring overnight at room temperature. After it was detected by TLC that the reaction was complete, the reaction was quenched by adding an appropriate amount of methanol at 0° C. The solution was concentrated and purified by silica gel column chromatography (n-hexane/ethyl acetate: 5:1.fwdarw.3:1) to obtain the compound 44* (128 mg, 76%). R.sub.f=0.50, Hexane/EtOAc=2:1. [α].sup.22.sub.D=−45.3 (c 1.0, CH.sub.3Cl). .sup.1H NMR (400 MHz, Chloroform-d) δ7.47-6.96 (m, 50H, arom. H), 5.67 (d, J=3.9 Hz, 1H, 1-H), 5.33 (d, J=6.9 Hz, 1H, NH), 5.13 (d, J=10.2 Hz, 1H, 1-H), 4.98 (d, J=3.9 Hz, 1H, 1-H), 4.85-4.71 (m, 5H), 4.70-4.63 (m, 4H), 4.64-4.55 (m, 5H), 4.55-4.45 (m, 6H), 4.50 (d, J=8.3 Hz, 1H, 1-H), 4.41 (d, J=2.7 Hz, 2H), 4.36 (d, J=10.6 Hz, 1H), 4.26 (q, J=6.4 Hz, 1H), 4.10-4.02 (m, 3H), 4.01-3.94 (m, 3H), 3.89 (d, J=11.3 Hz, 1H), 3.85-3.73 (m, 3H), 3.73-3.67 (m, 1H), 3.66-3.61 (m, 2H), 3.61-3.58 (m, 1H), 3.55 (dd, J=9.8, 3.1 Hz, 1H), 3.32 (dd, J=8.9, 4.9 Hz, 1H), 3.22 (dd, J=10.2, 3.0 Hz, 1H), 3.16 (d, J=2.4 Hz, 1H), 3.04 (q, J=9.2 Hz, 1H), 2.74-2.53 (m, 2H), 1.32-1.18 (m, 6H, CH.sub.3/CH.sub.3-Fucose), 1.11 (d, J=6.4 Hz, 3H, CH.sub.3-Fucose). .sup.13C NMR (101 MHz, Chloroform-d) δ153.3, 139.3, 138.8, 138.8, 138.8, 138.7, 138.5, 138.4, 138.3, 138.1, 137.7, 128.8, 128.6, 128.5, 128.5, 128.4, 128.4, 128.3, 128.2, 128.1, 128.0, 127.8, 127.7, 127.5, 127.5, 127.4, 127.3, 127.2, 127.2, 127.1, 126.1, 100.0, 98.5, 97.9, 95.3, 83.9, 82.3, 80.2, 80.1, 79.0, 78.2, 78.2, 75.8, 75.6, 75.5, 75.4, 74.8, 74.4, 74.3, 73.5, 73.5, 73.2, 72.9, 72.7, 72.4, 72.4, 72.3, 71.9, 70.9, 68.1, 67.7, 59.1, 24.3, 16.4, 16.2, 15.3. IR (film): v=2871, 1741, 1498, 1455, 1364, 1096, 822, 736, 697 cm.sup.−1. HRMS (ESI) m/z calcd for C.sub.99H.sub.112Cl.sub.3NONS [M+NH.sub.4].sup.+1769.6640, found 1769.6631.
[0137] Compound 45*: The donor tetrasaccharide 44* (20 mg, 0.0114 mmol) and the receptor 28* (12 mg, 0.0228 mmol) were dissolved in anhydrous CH.sub.2Cl.sub.2 (1.1 mL), and 4 Å molecular sieves and NIS (3 mg, 0.0137 mmol) were added. The reaction temperature was lowered to −15° C., and then TfOH (0.11 μL, 0.0011 mmol) was added dropwise under argon protection. The reaction was performed at −20° C. for 3 h. After it was detected by TLC that the reaction was complete, an appropriate amount of Et.sub.3N was added to quench the reaction. The solution was filtered, washed with a 10% Na.sub.2S.sub.2O.sub.3 solution until the solution was colorless, then sequentially washed with saturated NaHCO.sub.3 and a saturated saline solution, and dried with anhydrous Na.sub.2SO.sub.4. The solution was concentrated and purified by silica gel column chromatography (n-hexane/ethyl acetate: 6/1-4/1) to obtain the compound 45* (18 mg, 70%).
[0138] Compound 46*: The compound 30* (50 mg, 0.022 mmol) was dissolved in pyridine (1 mL), HF/pyridine (1 M, 0.22 mL) was added at 0° C., and then the reaction was performed under stirring at room temperature for 5 h. After it was detected by TLC that the reaction was complete, an appropriate amount of DCM was added for dissolution. Then the reaction solution was washed with saturated NaHCO.sub.3 and a saturated saline solution, dried with anhydrous Na.sub.2SO.sub.4, filtered, concentrated, and separated and purified by column chromatography to obtain corresponding hemiacetal.
[0139] The hemiacetal obtained above was dissolved in CH.sub.2Cl.sub.2 (2 mL), CCl.sub.3CN (16 μL, 0.11 mmol) and DBU (10 μL, 0.066 mmol) were added at 0° C., and the reaction was performed under stirring at room temperature for 2 h. After it was detected by TLC that the reaction was complete, the reaction solution was concentrated at 30° C., and then separated and purified by silica gel column chromatography (n-hexane/ethyl acetate: 5/1.fwdarw.3/1) to obtain the compound 46* (21 mg, 42%).
(2) Synthesis of target tridecasaccharide:
[0140] The synthetic routes are shown in
[0141] The implementation of the [5+8] synthesis strategy was carried out with 1.2 equivalent of glycosyl donor 43* or 46* and an octasaccharide receptor 27* under the action of an activating reagent TMSOTf. Unfortunately, the target molecule was not obtained, and the synthesis strategy [5+8] failed. Because considering that the reaction of the fully protected Le.sup.y tetrasaccharide with the galactose receptor 28* can be carried out successfully, the synthesis strategy of [4+9] was considered. 1.2 equivalent of glycosyl donor 41* or 44* reacted with the nonasaccharide receptor 49, the activating reagents were TfOH and NIS, and after the reaction was tried, the target molecule tridecasaccharide was still not obtained. Therefore, inferring the reasons for the failure of the above reaction strategy, the inventors have drawn two important conclusions: one is that the branched chain structure of the glycosylation donor increases the steric hindrance of the reaction, thereby leading to the failure of the reaction; and the second is the mismatch in activity between the glycosylation donor and the receptor, the donor activity is relatively high while the receptor activity is relatively low, thereby leading to the failure of the reaction. Therefore, the main solution strategy is to reduce the steric hindrance of the reaction and reduce the reactivity of the donor.