Chemical Synthesis Method of Plesiomonas Shigelloides Serotype O51 O-antigen Oligosaccharide
20190233459 ยท 2019-08-01
Inventors
Cpc classification
C07H15/04
CHEMISTRY; METALLURGY
Y02P20/55
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
C07H1/00
CHEMISTRY; METALLURGY
C07H15/06
CHEMISTRY; METALLURGY
C07H15/12
CHEMISTRY; METALLURGY
C07H13/04
CHEMISTRY; METALLURGY
International classification
C07H15/12
CHEMISTRY; METALLURGY
C07H15/04
CHEMISTRY; METALLURGY
C07H1/00
CHEMISTRY; METALLURGY
C07H13/04
CHEMISTRY; METALLURGY
Abstract
The present disclosure discloses the chemical synthesis method of the Plesiomonas shigelloides serotype O51 O-antigen oligosaccharide, belonging to the field of chemistry. Source-abundant D-glucose, L-fucose, D-glucosamine and the like are used as raw materials to prepare three glycosylation building blocks, the synthetic route composed of 11 reaction modules is designed, and through the optimization of protecting group and the optimization of the time of introducing functional group, the preparation of the target oligosaccharide chain is successfully achieved. The oligosaccharide chain prepared in the present disclosure has the advantages of cheap and easy-to-get raw materials, and simple and easy-to-repeat preparation method. The present disclosure will have good application prospects in the aspects of development of new drugs and vaccines of Plesiomonas shigelloides, and the like.
Claims
1. A linkered oligosaccharide chain fragment of the Plesiomonas shigelloides serotype O51 O-antigen polysaccharide, wherein the structure of the oligosaccharide chain fragment is expressed as the general Formula I:
V*[U.sub.x+2U.sub.x+1U.sub.x].sub.nVO-L-NH.sub.2Formula I, wherein X is 1, 2, 3; wherein n is 1, 2, 3; wherein V represents: the chemical bond, U.sub.x+2, or U.sub.x+2U.sub.x+1; wherein V* represents: H, HU.sub.x, or HU.sub.x+1U.sub.x; wherein L represents the linker; and U.sub.x, U.sub.x+1 and U.sub.x+2 are as shown in Formula V: ##STR00006##
2. The linkered oligosaccharide chain fragment of the Plesiomonas shigelloides serotype O51 O-antigen polysaccharide according to claim 1, wherein the structure of the oligosaccharide chain fragment is expressed as the general Formula II:
V*[U.sub.x+2U.sub.x+1U.sub.x].sub.nO-L-NH.sub.2Formula II, wherein x, n, L, U.sub.x, U.sub.x+1, U.sub.x+2 and V* are in accordance with the general Formula I; and the general Formula II is further expressed as the general Formulae II-a, II-b or II-c: ##STR00007##
3. The linkered oligosaccharide chain fragment of the Plesiomonas shigelloides serotype O51 O-antigen polysaccharide according to claim 1, wherein the structure of the oligosaccharide chain fragment is expressed as the general Formula III:
V*[U.sub.x+2U.sub.x+1U.sub.x].sub.nU.sub.x+2O-L-NH.sub.2Formula III, wherein x, n, L, U.sub.x, U.sub.x+1, U.sub.x+2 and V* are in accordance with the general Formula I, and the general Formula III is further expressed as the general Formulae III-a, III-b or III-c: ##STR00008##
4. The linkered oligosaccharide chain fragment of the Plesiomonas shigelloides serotype O51 O-antigen polysaccharide according to claim 1, wherein the structure of the oligosaccharide chain fragment is expressed as the general Formula IV:
V*[U.sub.x+2U.sub.x+1U.sub.x].sub.nU.sub.x+2U.sub.x+1O-L-NH.sub.2Formula IV, wherein x, n, L, U.sub.x, U.sub.x+1, U.sub.x+2 and V* are in accordance with the general Formula I, and the general Formula IV is further specifically expressed as general Formulae IV-a, IV-b and IV-c: ##STR00009##
5. A method of preparing the linkered oligosaccharide chain fragment of the Plesiomonas shigelloides serotype O51 O-antigen polysaccharide according to claim 1, wherein monosaccharide building block 1 is used as raw material of D-quinovosamine in an oligosaccharide chain, and the monosaccharide building block 1 is expressed as Formula VI-1: ##STR00010## wherein PG.sup.1 is acetyl, levulinyl, benzoyl, chloroacetyl, dichloroacetyl, trichloroacetyl, pivaloyl, allyloxycarbonyl, 2-naphthylmethyl, p-methoxybenzyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, or triethylsilyl; and wherein PG.sup.2 is benzyl; and LG is ethylthio, p-tolylthio, phenylthio, bromine, fluorine, trichloroacetimidate, N-phenyl trifluoroacetimidate or dibutyl phosphate.
6. The method according to claim 5, wherein monosaccharide building block 2 is used as raw material of L-fucosamine in the oligosaccharide chain, and the monosaccharide building block 2 is expressed as Formula VI-2: ##STR00011## wherein PG.sup.3 and PG.sup.4 are acetyl, levulinyl, benzoyl, chloroacetyl, dichloroacetyl, trichloroacetyl, pivaloyl, allyloxycarbonyl, 2-naphthylmethyl, p-methoxybenzyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, or triethylsilyl; and wherein LG is ethylthio, p-tolylthio, phenylthio, bromine, fluorine, trichloroacetimidate, N-phenyl trifluoroacetimidate or dibutyl phosphate.
7. The method according to claim 5, wherein monosaccharide building block 3 is used as raw material of 2,3-diamido-D-glucuronic acid in the oligosaccharide chain, and the monosaccharide building block 3 is expressed as Formula VI-3: ##STR00012## wherein PG.sup.5 is trichloroacetyl, dichloroacetyl or chloroacetyl; PG.sup.6 is benzyl, 2-naphthylmethyl, p-methoxybenzyl or levulinyl; PG.sup.7 is benzyl; and wherein LG is ethylthio, p-tolylthio, phenylthio, bromine, fluorine, trichloroacetimidate, N-phenyl trifluoroacetimidate or dibutyl phosphate.
8. The method according to claim 7, wherein the monosaccharide building block 3 is used as raw material of 2,3-diamido-D-glucuronic acid in the oligosaccharide chain, and the monosaccharide building block 3 is expressed as Formula VI-3: ##STR00013## wherein PG.sup.5 is trichloroacetyl, dichloroacetyl or chloroacetyl; wherein PG.sup.6 is benzyl, 2-naphthylmethyl, p-methoxybenzyl or levulinyl; wherein PG.sup.7 is benzyl; and wherein LG is ethylthio, p-tolylthio, phenylthio, bromine, fluorine, trichloroacetimidate, N-phenyl trifluoroacetimidate or dibutyl phosphate.
9. The method according to claim 5, wherein a compound of VI-4 is assembled as a linker raw material to the reducing end of the oligosaccharide chain, and wherein PG.sup.9 and PG.sup.10 are benzyl or benzyloxycarbonyl; ##STR00014##
10. The method according to claim 5, further comprises preparing important intermediate 3-azidoglycosamine for the monosaccharide building block 3, and wherein when performing double inversion at 3-position, 1) 3-trifluoromethanesulfonyl glucosamine is subjected to the Lattrell-Dax reaction to obtain 3-hydroxyl allosamine; 2) the 3-hydroxyl allosamine is subjected to trifluoromethanesulfonylation to obtain 3-trifluoromethanesulfonyl allosamine; and 3) the 3-trifluoromethanesulfonyl allosamine is subjected to azido nucleophilic substitution to obtain the 3-azidoglycosamine.
11. The method according to claim 5, comprising 11 reaction modules in turn, wherein the 11 reaction modules are: A: glycosylation reaction; B: azido reduction and acetylation; C: quinovosamine 3-position deprotection; D: azido reduction and protection by 2,2,2-trichloroethoxycarbonyl (Troc); E: fucosamine 3-/4-position selective naphthylmethyl protection; F: fucosamine 3-position acetylation; G: azido reduction and butyrylation; H: removal of Troc and modification to acetamidine; I: hydrogenation global deprotection; J: glucuronic acid 4-position deprotection; and K: azido reduction and modification to acetamidine.
12. The method according to claim 5, comprising 11 reaction modules in turn, wherein the 11 reaction modules are: A: glycosylation reaction; B: azido reduction and acetylation; C: quinovosamine 3-position deprotection; D: azido reduction and protection by 2,2,2-trichloroethoxycarbonyl (Troc); E: fucosamine 3-/4-position selective naphthylmethyl protection; F: fucosamine 3-position acetylation; G: azido reduction and butyrylation; H: removal of Troc and modification to acetamidine; I: hydrogenation global deprotection; J: glucuronic acid 4-position deprotection; and K: azido reduction and modification to acetamidine.
13. The method according to claim 5, comprising 11 reaction modules in turn, wherein the 11 reaction modules are: A: glycosylation reaction; B: azido reduction and acetylation; C: quinovosamine 3-position deprotection; D: azido reduction and protection by 2,2,2-trichloroethoxycarbonyl (Troc); E: fucosamine 3-/4-position selective naphthylmethyl protection; F: fucosamine 3-position acetylation; G: azido reduction and butyrylation; H: removal of Troc and modification to acetamidine; I: hydrogenation global deprotection; J: glucuronic acid 4-position deprotection; and K: azido reduction and modification to acetamidine.
14. The method according to claim 5, comprising 11 reaction modules in turn, wherein the 11 reaction modules including: A: glycosylation reaction; B: azido reduction and acetylation; C: quinovosamine 3-position deprotection; D: azido reduction and protection by 2,2,2-trichloroethoxycarbonyl (Troc); E: fucosamine 3-/4-position selective naphthylmethyl protection; F: fucosamine 3-position acetylation; G: azido reduction and butyrylation; H: removal of Troc and modification to acetamidine; I: hydrogenation global deprotection; J: glucuronic acid 4-position deprotection; and K: azido reduction and modification to acetamidine.
15. A Plesiomonas shigelloides vaccine, comprising the linkered oligosaccharide chain fragment of the Plesiomonas shigelloides serotype O51 O-antigen polysaccharide according to claim 1, which services as the epitope; or, comprising the linked oligosaccharide chain fragment of the Plesiomonas shigelloides serotype O51 O-antigen polysaccharide according to claim 2, which services as the epitope; or comprising the linked oligosaccharide chain fragment of the Plesiomonas shigelloides serotype O51 O-antigen polysaccharide according to claim 3, which services as the epitope; or comprising the linked oligosaccharide chain fragment of the Plesiomonas shigelloides serotype O51 O-antigen polysaccharide according to claim 4, which services as the epitope.
Description
BRIEF DESCRIPTION OF FIGURES
[0095]
[0096]
[0097]
[0098]
[0099]
[0100]
[0101]
[0102]
[0103]
[0104]
[0105]
[0106]
[0107]
[0108]
[0109]
[0110]
[0111]
[0112]
[0113]
[0114]
[0115]
[0116]
[0117]
DETAILED DESCRIPTION
[0118] Commercially available reagents used in experiments were directly used without any treatment. The anhydrous solvents used in reaction were prepared by an MBraun MB-SPS 800 solvent drying system. Solvents used in silica gel column chromatography were all analytically pure and used after distillation under reduced pressure. A silica gel plate used for thin layer chromatography (TLC) was a glass-based or aluminum foil-based silica gel plate prepared from 60-F254 silica gel. A thin layer chromatography stain was a sugar stain (0.1% (v/v) 3-methoxyphenol, 2.5% (v/v) sulfuric acid ethanol solution), or a CAM stain (5% (w/v) ammonium molybdate, 1% (w/v) cerium sulfate (II) and 10% (v/v) sulfuric acid aqueous solution), or a ninhydrin stain (1.5% (w/v) ninhydrin and 3% (v/v) acetic acid n-butanol solution). Silica gel used for normal-phase silica gel column chromatography was 200-300-mesh silica gel. A column used in the molecular exclusion chromatography was Sephadex LH-20 (GE Healthcare).
[0119] The yield of each reaction step was calculated separately, and the yield was calculated according to: (amount of target product/amount of raw materials)100%. The structure of the product was identified by nuclear magnetic spectroscopy, infrared spectroscopy, optical rotation and high-resolution mass spectrometry. The purity of the product was analyzed by nuclear magnetic spectroscopy and high performance liquid chromatography. The .sup.1H, .sup.13C and two-dimensional nuclear magnetic spectrum were measured by a Bruker Ascend 400 MHz nuclear magnetic resonance spectrometer, or a Bruker Ultrashield Plus 400 MHz nuclear magnetic resonance apparatus, or a Bruker AVIII 700 MHz nuclear magnetic resonance spectrometer at 25 C. The high resolution mass spectrum was measured by an Agilent 6220 electrospray ionization time-of-flight mass spectrometer. The optical rotation was measured by the Schmidt & Haensch UniPol L 1000 automatic polarimeter at 589 nm, and the concentration (c) unit was g/100 mL. The infrared spectrum was measured by the Thermo Fisher Scientific Nicolet iS5 infrared meter. The analytical high performance liquid chromatography was performed by an Agilent 1200 series liquid chromatograph-quadrupole electrospray mass spectrometer 6130 using the Thermo Scientific Hypercarb column (1504.6 mm) as an analytical column. The preparative high performance liquid chromatography was performed by an Agilent 1200 series liquid-phase chromatograph-quadrupole electrospray mass spectrometer 6130 using the Thermo Scientific Hypercarb column (15010 mm) as the semi-preparative column.
Embodiment 1
Synthesis of allyl 3,4,6-tri-O-acetyl-2-deoxy-2-trichloroacetamido--D-glucopyranose (1*)
[0120] A reaction equation is as shown in
[0121] Under argon protection, 1,3,4,6-tetra-O-acetyl-2-deoxy-2-trichloroacetamido--D-glucopyranose (M. Virlouvet et al., Adv. Synth. Catal. 2010, 352, 2657-2662) (27 g, 0.055 mol) was dissolved in anhydrous dichloromethane (130 mL), and an activated 4 molecular sieve and allyl alcohol (18.7 mL, 0.274 mol) were added. After cooling to 5 C., boron trifluoride diethyl ether complex (70 mL, 0.548 mol) was added dropwise. The reaction solution was stirred at 0 C. for 30 min, and then was heated to room temperature to reaction for 71 hours. After the completion of the reaction, the reaction solution was poured onto 200 g of crushed ice, an organic phase obtained by filtration through celite was respectively extracted with water, the saturated sodium bicarbonate solution and the saturated saline solution, and the resulting organic phase was dried with sodium sulfate and then concentrated. The resulting crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate, 5:1, v/v) to obtain the yellow syrup-like pure product 1* (19.9 g, 0.041 mol, 74%). [].sub.D.sup.20=+87.8 (c=1.20, CHCl.sub.3); IR v.sub.max (film) 3429, 2960, 1749, 1722, 1517, 1368, 1229, 1048, 822, 682 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =6.93 (d, J=9.1 Hz, 1H, NH), 5.88 (dddd, J=17.0, 10.3, 6.5, 5.4 Hz, 1H, CHC), 5.41-5.23 (m, 3H, 4-H, CCH.sub.2), 5.16 (dd, J=9.8, 9.8 Hz, 1H, 3-H), 5.01 (d, J=3.7 Hz, 1H, 1-H), 4.34-4.19 (m, 3H, 6-CH.sub.2, OCH.sub.a), 4.11 (dd, J=12.4, 2.4 Hz, 1H, OCH.sub.b), 4.09-3.98 (m, 2H, 2-H, 5-H), 2.11 (s, 3H, CH.sub.3CO), 2.05 (s, 3H, CH.sub.3CO), 2.02 (s, 3H, CH.sub.3CO); .sup.13C NMR (100 MHz, CDCl.sub.3) =171.1, 170.6, 169.3, 161.9, 132.6, 119.1, 95.5, 92.1, 77.3, 70.7, 69.0, 68.1, 67.8, 61.8, 53.9, 20.7, 20.63, 20.59; HR-ESI-MS (m/z): calcd for C.sub.17H.sub.22Cl.sub.3NO.sub.9Na.sup.+(M+Na.sup.+): 512.0258, found: 512.0254.
Embodiment 2
Synthesis of allyl 4,6-O-benzylidene-2-deoxy-2-trichloroacetamido--D-glucopyranose (2*)
[0122] A reaction equation is as shown in
[0123] Compound 1* (50 g, 0.102 mol) was dissolved in methanol (800 mL), sodium methoxide (2.8 g, 0.051 mol) was added. And then the mixture was stirred at room temperature for 5 hours. After completion of the reaction, the reaction solution was neutralized with an Amberlite IR 120 cation exchange resin, filtered, and concentrated to give desired white solid-like 3,4,6-trihydroxy saccharide (37.1 g, 0.102 mol, quant.).
[0124] The trihydroxy saccharide (71 g, 0.195 mol) was further dissolved in 300 mL of anhydrous DMF, and benzaldehyde dimethyl acetal (35 mL, 0.234 mol) and p-toluenesulfonic acid (4.45 g, 0.023 mol) were added. After the reaction solution reacted at 60 C. for 24 hours, methanol generated in the reaction was distilled off under reduced pressure by the rotary evaporator, and the reaction was further carried out at 60 C. This operation was repeated, and DMF was distilled off under reduced pressure after the reaction was completed. The resulting crude product was dissolved in ethyl acetate, extracted with the saturated sodium bicarbonate solution, water and the saturated saline solution, dried with anhydrous sodium sulfate and evaporated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate, 10:1, v/v) to obtain the white solid-like product 2* (73.6 g, 0.163 mol, 83%). [].sub.D.sup.20=+68.6 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3339, 2919, 1695, 1530, 1451, 1372, 1085, 1027, 989, 748, 697 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.55-7.34 (m, 5H, Ph), 6.96 (d, J=8.8 Hz, 1H, NH), 5.86 (dddd, J=16.9, 10.3, 6.4, 5.3 Hz, 1H, CHC), 5.53 (s, 1H, PhCH), 5.35-5.20 (m, 2H, CCH.sub.2), 4.97 (d, J=3.8 Hz, 1H, 1-H), 4.27 (dd, J=10.2, 4.8 Hz, 1H, 4-H), 4.24-4.11 (m, 2H, 2-H, OCH.sub.a), 4.06-3.94 (m, 2H, OCH.sub.b, 5-H), 3.86 (ddd, J=9.9, 9.8, 4.7 Hz, 1H, 6-CHa), 3.74 (t, J=10.3 Hz, 1H, 6-CH.sub.b), 3.56 (t, J=9.3 Hz, 1H, 3-H), 2.78 (br, 1H, 3-OH); .sup.13C NMR (100 MHz, CDCl.sub.3) =162.3, 137.0, 132.9, 129.3, 128.3, 126.3, 118.7, 101.9, 96.3, 92.4, 81.5, 69.7, 68.8, 68.6, 62.8, 55.4; HR-ESI-MS (m/z): calcd for C.sub.18H.sub.20Cl.sub.3NO.sub.6Na.sup.+ (M+Na.sup.+): 474.0254, found: 474.0245.
Embodiment 3
Synthesis of allyl 4,6-O-benzylidene-3-O-trifluoromethanesulfonyl-2-trichloroacetamido-2-deoxy--D-glucopyranose (3*)
[0125] A reaction equation is as shown in
[0126] Compound 2* (53.1 g, 0.117 mol) was dissolved in an anhydrous dichloromethane/pyridine mixture (660 mL, 7:1, v/v), cooled to 20 C., followed by dropwise addition of the dichloromethane solution (150 mL) of trifluoromethanesulfonic anhydride (40 mL, 0.234 mol) with stirring, and gradually warmed to 10 C. within 2 hours. After completion of the reaction, the reaction solution was diluted with dichloromethane, and sequentially extracted with 1 M HCl solution, saturated sodium bicarbonate, water and the saturated saline solution. An organic phase was dried with anhydrous sodium sulfate and subjected to reduced pressure distillation at low temperature to remove the solvent. The resulting crude product was purified by silica gel column chromatography (ethyl ether:ethyl acetate, 5:1 to 2:1, v/v) to obtain the yellow syrup-like product 3* (66.5 g, 0.114 mol, 98%). [].sub.D.sup.20=+35.3 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3348, 1698, 1529, 1414, 1373, 1203, 1146, 1122, 959, 836, 753, 628 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.52-7.33 (m, 5H, Ph), 7.06 (d, J=9.6 Hz, 1H, NH), 5.87 (dddd, J=17.0, 10.2, 6.8, 5.4 Hz, 1H, CHC), 5.62 (s, 1H, PhCH), 5.37-5.27 (m, 2H, CCH.sub.2), 5.13 (dd, J=10.3, 9.1 Hz, 1H, 3-H), 5.03 (d, J=3.7 Hz, 1H, 1-H), 4.50 (ddd, J=10.0, 10.0, 3.8 Hz, 1H, 2-H), 4.36 (dd, J=10.4, 4.5 Hz, 1H, 6-CHa), 4.25 (ddt, J=12.7, 5.5, 1.3 Hz, 1H, OCH.sub.a), 4.06 (ddt, J=12.7, 6.8, 1.2 Hz, 1H, OCH.sub.b), 3.98 (td, J=9.6, 4.6 Hz, 1H, 5-H), 3.92 (dd, J=9.2 Hz, 1H, 6-CH.sub.b), 3.84 (dd, J=10.1, 10.1 Hz, 1H, 4-H); .sup.13C NMR (100 MHz, CDCl.sub.3) =162.2, 136.2, 132.2, 129.2, 128.2, 125.9, 119.9, 116.8, 101.6, 96.4, 91.8, 83.1, 78.2, 69.3, 68.4, 63.3, 53.8; HR-ESI-MS (m/z): calcd for C.sub.19H.sub.19Cl.sub.3F.sub.3NO.sub.8SNa.sup.+(M+Na.sup.+): 605.9747, found: 605.9760.
Embodiment 4
Synthesis of allyl 4,6-O-benzylidene-2-trichloroacetamido-2-deoxy--D-allopyranose (4*)
[0127] A reaction equation is as shown in
[0128] Compound 3* (0.5 g, 0.855 mmol) was dissolved in anhydrous DMF (10 mL), potassium nitrite (364 mg, 4.275 mmol) was added, and the reaction solution was stirred at 50 C. for 6 hours. After the reaction was completed, the reaction solution was diluted by adding dichloromethane, and extracted with the saturated saline solution. And the resulting organic phase was dried with anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (petroleum ether:ethyl acetate, 8:1, v/v) to give the white solid-like 3-position inversion product 4* (278 mg, 0.614 mmol, 72%). [].sub.D.sup.20=+74.0 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3419, 1711, 1507, 1378, 1218, 1102, 1065, 1023, 820, 756, 699 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.57-7.33 (m, 6H, NH, Ph), 5.88 (dddd, J=16.9, 10.3, 6.4, 5.2 Hz, 1H, CHC), 5.63 (s, 1H, PhCH), 5.37-5.20 (m, 2H, CCH.sub.2), 5.00 (d, J=4.2 Hz, 1H, 1-H), 4.38 (dd, J=10.3, 5.1 Hz, 1H, 6-CH.sub.b), 4.27 (m, 2H, 3-H, OCH.sub.a), 4.24-4.16 (m, 2H, 2-H, 5-H), 4.05 (ddt, J=13.0, 6.5, 1.3 Hz, 1H, OCH.sub.b), 3.80 (t, J=10.3 Hz, 1H, 6-CHa), 3.67 (dd, J=9.7, 2.8 Hz, 1H, 4-H), 2.66 (br, 1H, 3-OH); .sup.13C NMR (100 MHz, CDCl.sub.3) =161.7, 136.9, 132.9, 129.3, 128.4, 126.2, 118.8, 101.9, 96.0, 92.3, 78.1, 77.2, 69.5, 69.0, 67.3, 57.6, 51.2; HR-ESI-MS (m/z): calcd for C.sub.18H.sub.20C.sub.3NO.sub.6Na.sup.+(M+Na.sup.+): 474.0254, found: 474.0245.
Embodiment 5
Synthesis of allyl 4,6-O-benzylidene-3-O-trifluoromethanesulfonyl-2-trichloroacetamido-2-deoxy--D-allopyranose (5*)
[0129] A reaction equation is as shown in
[0130] Pyridine (273 L, 3.380 mmol) was added to an anhydrous dichloromethane (3.93 mL) solution of compound 4* (178 mg, 0.393 mmol) at 20 C., trifluoromethanesulfonic anhydride (133 L, 0.786 mmol) was further added dropwise, and the reaction temperature was gradually increased to 10 C. during 2 hours of stirring. After the reaction was completed, the reaction solution was diluted with dichloromethane, and the resulting organic phase was extracted with a 1 M HCl solution, the saturated sodium bicarbonate solution, water and the saturated saline solution. The resulting organic phase was dried with anhydrous sodium sulfate and subjected to reduced pressure distillation at low temperature to remove the solvent. The resulting crude product was purified by the silica gel column (petroleum ether:ethyl acetate, 10:1-5:1-2:1, v/v) to give the yellow syrup-like product 5* (160 mg, 0.275 mmol, 70%). [].sub.D.sup.20=+40.4 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 2923, 2853, 1750, 1497, 1416, 1211, 1028, 821, 617 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.53-7.35 (m, 5H, Ph), 7.33 (d, J=8.5 Hz, 1H, NH), 5.89 (dddd, J=16.8, 10.6, 6.0, 4.7 Hz, 1H, CHC), 5.61 (s, 1H, PhCH), 5.44 (t, J=2.9 Hz, 1H, 3-H), 5.37 (dq, J=17.3, 1.6 Hz, 1H, CCHa), 5.28 (dq, J=10.5, 1.4 Hz, 1H, CCH.sub.b), 4.96 (d, J=4.3 Hz, 1H, 1-H), 4.44 (ddd, J=8.0, 4.3, 3.2 Hz, 1H, 2-H), 4.41-4.31 (m, 2H, 6-Ha, OCH.sub.a), 4.26 (td, J=9.9, 5.2 Hz, 1H, 5-H), 4.05 (ddt, J=13.3, 6.0, 1.4 Hz, 1H, OCH.sub.b), 3.87 (dd, J=9.7, 2.5 Hz, 1H, 4-H), 3.78 (t, J=10.4 Hz, 1H, 6-Hb); .sup.13C NMR (100 MHz, CDCl.sub.3) =162.0, 136.3, 132.4, 129.4, 128.3, 126.2, 118.1, 102.4, 94.9, 91.5, 80.7, 74.9, 69.0, 68.8, 58.1, 50.2; HR-ESI-MS (m/z): calcd for C.sub.19H.sub.19Cl.sub.3F.sub.3NO.sub.8SNa.sup.+(M+Na.sup.+): 605.9747, found: 605.9753.
Embodiment 6
Synthesis of allyl 4,6-O-benzylidene-3-azido-2-trichloroacetamido-2,3-dideoxy--D-glucopyranose (6*)
[0131] A reaction equation is as shown in
[0132] Compound 5* (65 mg, 0.111 mmol) was dissolved in anhydrous DMF, and sodium azide (36 mg, 0.555 mmol) was added and stirred at room temperature overnight. After the completion of the reaction, the reaction solution was diluted with ethyl acetate and extracted with water and the saturated saline solution. The resulting organic phase was dried with anhydrous sodium sulfate and concentrated. The resulting crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate, 20:1-8:1, v/v) to obtain the white solid-like product 6* (44.2 mg, 0.093 mmol, 83%). [].sub.D.sup.20=+39.7 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3120, 2917, 2849, 2110, 1692, 1528, 1372, 1258, 1124, 1080, 1055, 1016, 836, 750, 697 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.55-7.33 (m, 5H, Ph), 6.88 (d, J=9.5 Hz, 1H, NH), 5.94-5.80 (m, 1H, CHC), 5.63 (s, 1H, PhCH), 5.36-5.23 (m, 2H, CCH.sub.2), 4.92 (d, J=3.6 Hz, 1H, 1-H), 4.32 (dd, J=10.4, 4.9 Hz, 1H, 6-CHa), 4.24 (ddt, J=12.7, 5.5, 1.4 Hz, 1H, OCH.sub.a), 4.13 (ddd, J=11.2, 9.7, 3.7 Hz, 1H, 2-H), 4.05 (ddt, J=12.8, 6.6, 1.4 Hz, 1H, OCH.sub.b), 3.99-3.88 (m, 2H, 3-H, 5-H), 3.79 (t, J=10.3 Hz, 1H, 6-CH.sub.b), 3.70 (t, J=9.6 Hz, 1H, 4-H); .sup.13C NMR (100 MHz, CDCl.sub.3) =161.9, 136.7, 132.6, 129.1, 128.3, 125.9, 119.2, 101.5, 95.9, 92.3, 80.3, 77.2, 68.9, 68.7, 63.3, 61.2, 53.5; HR-ESI-MS (m/z): calcd for C.sub.18H.sub.19Cl.sub.3N.sub.4O.sub.5Na.sup.+(M+Na.sup.+): 499.0319, found: 499.0311.
Embodiment 7
Synthesis of allyl 3-azido-2-trichloroacetamido-2,3-dideoxy--D-glucopyranose (7*)
[0133] A reaction equation is as shown in
[0134] Compound 6* (20 mg, 0.042 mmol) was added to an 80% acetic acid aqueous solution (0.9 mL), heated to 55 C., and stirred until the reaction was completed. The reaction solution was subjected to reduced pressure distillation to remove the solvent, and purified by silica gel column chromatography (dichloromethane:methanol, 60:1-40:1-30:1, v/v) to obtain white solid-like 4,6-dihydroxy saccharide 7* (16.3 mg, 0.042 mmol, quant.). [].sub.D.sup.20=+16.9 (c=0.35, CHCl.sub.3); IR v.sub.max (film) 3411, 2924, 2108, 1712, 1517, 1262, 1049, 822, 680 cm.sup.1; .sup.1H NMR (400 MHz, CD.sub.3OD) =5.98 (dddd, J=17.0, 10.4, 6.4, 5.1 Hz, 1H, CHC), 5.38 (dq, J=17.3, 1.7 Hz, 1H, CCHa), 5.24 (dq, J=10.4, 1.4 Hz, 1H, CCH.sub.b), 4.94 (d, J=3.6 Hz, 1H, 1-H), 4.29 (ddt, J=13.2, 5.2, 1.5 Hz, 1H, OCH.sub.a), 4.10 (ddt, J=13.1, 6.4, 1.4 Hz, 1H, OCH.sub.b), 3.99 (dd, J=11.5, 9.2 Hz, 1H, 3-H), 3.93-3.81 (m, 2H, 2-H, 6-Ha), 3.81-3.68 (m, 2H, 5-H, 6-Hb), 3.52 (t, J=9.3 Hz, 1H, 4-H); .sup.13C NMR (100 MHz, CD.sub.3OD) =166.7, 137.6, 120.8, 98.9, 76.5, 73.7, 72.0, 67.5, 64.8, 58.1; HR-ESI-MS (m/z): calcd for C.sub.11H.sub.15Cl.sub.3N.sub.4O.sub.5Na.sup.+(M+Na.sup.+): 411.0006, found: 411.0001.
Embodiment 8
Synthesis of benzyl (allyl 3-azido-2,3-dideoxy-2-trichloroacetamido--D-glucopyranosid) uronate (8*)
[0135] A reaction equation is as shown in
[0136] At 0 C., water (1.4 L), 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO) (4.4 g, 0.028 mol) and diacetoxyliodo benzene (BAIB) (45.5 g, 0.141 mol) were added to the dichloromethane (2.8 L) solution of compound 7* (22 g, 0.056 mol). After the reaction solution was stirred at room temperature for 4 hours. After that, the reaction mixture was passed through a pad of silica gel, concentrated and dried under high vacuum. The crude acid was used in the next step without further purification.
[0137] The crude carboxylic acid compound was dissolved in anhydrous DMF (2.8 L), sodium bicarbonate (21.4 g, 0.255 mol) and benzyl bromide (50.4 mL, 0.424 mol) were sequentially added, and the resulting reaction solution was stirred at room temperature. After the completion of the reaction, the solvent was distilled off under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate, 10:1, v/v) to give the yellow syrup-like benzylglucuronate 8* (20.4 g, 0.041 mol, yield of 73% in two-step reaction). [].sub.D.sup.20=+54.5 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3412, 2930, 2110, 1717, 1514, 1264, 1057, 909, 820, 733, 698 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.43-7.33 (m, 5H, Ph), 6.79 (d, J=9.4 Hz, 1H, NH), 5.86 (dddd, J=17.0, 10.3, 6.6, 5.4 Hz, 1H, CHC), 5.37-5.20 (m, 4H, PhCH.sub.2, CCH.sub.2), 4.97 (d, J=3.6 Hz, 1H, 1-H), 4.31-4.20 (m, 2H, 5-H, OCH.sub.a), 4.13-4.00 (m, 2H, 2-H, OCH.sub.b), 3.91 (t, J=9.5 Hz, 1H, 4-H), 3.75 (dd, J=11.1, 9.3 Hz, 1H, 3-H), 3.31 (br, 1H, 4-OH); .sup.13C NMR (100 MHz, CDCl.sub.3) =169.5, 161.9, 134.8, 132.4, 128.77, 128.75, 128.3, 119.4, 95.6, 92.2, 77.2, 71.2, 70.5, 69.3, 67.8, 63.6, 52.7; HR-ESI-MS (m/z): calcd for C.sub.18H.sub.19Cl.sub.3N.sub.4O.sub.6Na.sup.+(M+Na.sup.+): 515.0268, found: 515.0264.
Embodiment 9
Synthesis of benzyl (allyl 3-azido-4-O-benzyl-2,3-dideoxy-2-trichloroacetamido--D-glucopyranosid) uronate (9*)
[0138] A reaction equation is as shown in
[0139] Compound 8* (20.3 g, 0.041 mol) was dissolved in anhydrous dichloromethane (400 mL), and benzyl bromide (49 mL, 0.411 mol) and silver oxide (28.6 g, 0.123 mol) were added sequentially at 0 C. The resulting reaction solution was stirred at 0 C. for 5 hours, and then warmed to room temperature, stirred for 28 hours. After the completion of the reaction was confirmed, the mixture was filtered through celite and then concentrated. The resulting crude product was subjected to silica gel column chromatography (petroleum ether:ethyl acetate, 40:1-10:1, v/v) to obtain the white solid-like product 9* (14.7 g, 0.025 mol, 62%). [].sub.D.sup.20=+66.3 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3357, 2935, 2108, 1743, 1713, 1514, 1253, 1185, 1111, 1063, 1028, 820, 750, 697 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.43-7.10 (m, 10H, 2Ph), 6.81 (d, J=9.6 Hz, 1H, NH), 5.84 (dddd, J=17.0, 10.3, 6.6, 5.4 Hz, 1H, CHC), 5.32-5.22 (m, 2H, CCH.sub.2), 5.21 (s, 2H, COOCH.sub.2Ph), 4.95 (d, J=3.5 Hz, 1H, 1-H), 4.69 (d, J=10.5 Hz, 1H, PhCH.sub.2), 4.49 (d, J=10.5 Hz, 1H, PhCH.sub.2), 4.35-4.27 (m, 1H, 5-H), 4.24 (ddt, J=12.8, 5.4, 1.4 Hz, 1H, OCH.sub.a), 4.17-4.08 (m, 1H, 2-H), 4.04 (ddt, J=12.8, 6.6, 1.2 Hz, 1H, OCH.sub.b), 3.83-3.74 (m, 2H, 3-H, 4-H); .sup.13C NMR (100 MHz, CDCl.sub.3) =168.4, 161.8, 136.9, 134.9, 132.4, 128.7, 128.59, 128.56, 128.5, 128.4, 128.24, 128.21, 128.14, 128.12, 119.44, 119.39, 95.5, 92.3, 78.2, 75.1, 71.0, 69.1, 67.6, 64.3, 53.0; HR-ESI-MS (m/z): calcd for C.sub.25H.sub.25Cl.sub.3N.sub.4O.sub.6Na.sup.+(M+Na.sup.+): 605.0737, found: 605.0731.
Embodiment 10
Synthesis of benzyl 3-azido-4-O-benzyl-2,3-dideoxy-2-trichloroacetamido-D-glucopyranosyl uronate (10*)
[0140] A reaction equation is as shown in
[0141] Compound 9* (95.8 mg, 0.164 mmol) and acetic acid (28 L, 0.492 mmol) were dissolved in 1,4-dioxane (2 mL), and then selenium dioxide (91 mg, 0.820 mmol) was added and heated to reflux temperature. After the reaction was completed, the mixture was cooled to room temperature, and triethylamine was added dropwise to quench the reaction. The crude product obtained by concentration was purified by silica gel column chromatography (petroleum ether:ethyl acetate, 8:1-4:1, v/v) to obtain the light yellow solid-like hemiacetal product 10* (58.0 mg, 0.107 mmol, 65%). [].sub.D.sup.20=+33.2 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3410, 2944, 2113, 1712, 1518, 1457, 1357, 1264, 1187, 1113, 1068, 822, 752, 698 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.47-7.06 (m, 10H, 2Ph), 6.92 (d, J=9.5 Hz, 1H, NH), 5.29 (t, J=3.5 Hz, 1H, 1-H), 5.21 (d, J=12.1 Hz, 1H, PhCH.sub.a1), 5.17 (d, J=12.1 Hz, 1H, PhCH.sub.a2), 4.70 (d, J=10.5 Hz, 1H, PhCH.sub.b1), 4.56-4.44 (m, 2H, PhCHb.sub.2, 5-H), 4.11 (td, J=9.7, 3.4 Hz, 1H, 2-H), 3.81 (m, 2H, 3-H, 4-H), 3.57 (d, J=3.7 Hz, 1H, 1-OH).; .sup.13C NMR (100 MHz, CDCl.sub.3) =168.6, 162.1, 136.8, 134.7, 128.8, 128.7, 128.5, 128.3, 128.2, 92.2, 90.9, 78.0, 75.0, 70.8, 67.8, 63.7, 53.1; HR-ESI-MS (m/z): calcd for C.sub.22H.sub.21Cl.sub.3N.sub.4O.sub.6Na.sup.+(M+Na.sup.+): 567.0395, found: 567.0414.
Embodiment 11
Synthesis of benzyl (3-azido-4-O-benzyl-2,3-dideoxy-2-trichloroacetamido-D-glucopyranosyl 1-(N-phenyl)-2,2,2-trifluoroacetimidate-) uronate (11*)
[0142] A reaction equation is as shown in
[0143] Potassium carbonate (12 mg, 0.084 mmol) and N-phenyl trifluoroacetimidoyl chloride (13 L, 0.084 mmol) were added to an anhydrous dichloromethane (0.4 mL) solution of compound 10* (22.7 mg, 0.042 mmol), and the resulting reaction solution was stirred at room temperature overnight. After the reaction was completed, the organic phase was obtained by filtration, and the crude product obtained after concentration was purified by silica gel column chromatography (n-hexane:ethyl acetate, 18:1, v/v) to give the colorless syrup-like product 11* (27.4 mg, 0.038 mmol, 91%). IR v.sub.max (film) 3346, 2112, 1723, 1524, 1317, 1213, 1166, 1090, 823, 752, 697 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.44 (s, 1H), 7.35-7.11 (m, 20H), 7.03 (q, J=7.5 Hz, 2H), 6.80 (d, J=8.8 Hz, 1H), 6.73 (d, J=7.8 Hz, 2H), 6.68 (d, J=7.8 Hz, 2H), 6.38 (s, 1H), 6.12 (s, 1H), 5.30-5.02 (m, 3H), 4.61 (d, J=10.5 Hz, 1H), 4.56 (d, J=10.6 Hz, 1H), 4.52-4.44 (m, 2H), 4.37 (d, J=7.6 Hz, 1H), 4.24 (s, 1H), 4.07 (dt, J=14.0, 7.2 Hz, 1H), 3.92 (s, 1H), 3.85 (dd, J=13.4, 7.8 Hz, 2H); .sup.13C NMR (100 MHz, CDCl.sub.3) =167.4, 167.2, 162.0, 142.5, 136.2, 136.1, 134.7, 134.5, 128.83, 128.77, 128.69, 128.67, 128.64, 128.58, 128.56, 128.5, 128.41, 128.37, 124.8, 124.5, 119.3, 119.1, 75.0, 74.3, 73.8, 72.7, 67.9, 67.8, 62.5, 51.7; HR-ESI-MS (m/z): calcd for C.sub.30H.sub.25Cl.sub.3F.sub.3N.sub.5O.sub.6Na.sup.+(M+Na.sup.+): 738.0691, found: 738.0722.
Embodiment 12
Synthesis of ethyl 2-azido-3,4-di-O-acetyl-2-deoxy-1-thio-L-fucopyranoside (12*)
[0144] A reaction equation is as shown in
[0145] 2-azido-3,4-di-O-acetyl-2-deoxy-L-fucopyra nose (C. L. Pereira et al., Angew. Chem. Int. Ed. 2015, 54, 10016-10019) (44.4 mg, 0.16 mmol) was dissolved in an anhydrous dichloromethane/pyridine mixture (1.6 mL, 4:1, v/v). After cooling to 0 C., acetic anhydride (150 L, 1.6 mmol) was added dropwise. After the catalytic amount of dimethylaminopyridine was added, the reaction solution was stirred at room temperature. After the completion of the reaction was confirmed, the reaction solution was extracted with the saturated sodium bicarbonate solution, and the resulting organic phase was dried with anhydrous sodium sulfate and concentrated. The resulting crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate, 10:1, v/v) to obtain 1-O-acetyl saccharide (50.5 mg, 0.16 mmol, quant.).
[0146] 1-O-acetyl fucose (50.5 mg, 0.16 mmol) and ethanethiol (18 L, 0.242 mmol) were dissolved in anhydrous dichloromethane (1.6 mL), and an activated 4 molecular sieve was added and stirred for 30 minutes. After cooling to 10 C., trimethylsilyl trifluoromethanesulfonate (35 L, 0.193 mmol) was added, and the resulting reaction solution was stirred at 0 C. After the completion of the reaction was confirmed, the reaction was quenched by the addition of triethylamine, and the solvent is removed by reduced pressure distillation, and the resulting crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate, 10:1, v/v) to obtain the product 12* (44.0 mg, 0.14 mmol, 88%, :
=1:0.7). .sup.1H NMR (400 MHz, CDCl.sub.3) =5.45 (d, J=5.6 Hz, 1H, 1
-H), 5.28 (dd, J=3.4, 1.2 Hz, 1H, 4
-H), 5.26-5.21 (m, 1H, 4-H), 5.13 (dd, J=11.0, 3.3 Hz, 1H, 3
-H), 4.88 (dd, J=10.2, 3.3 Hz, 1H, 3
-H), 4.49 (qd, J=6.5, 1.3 Hz, 1H, 5
-H), 4.37 (d, J=10.2 Hz, 1H, 1-H), 4.22 (dd, J=11.0, 5.5 Hz, 1H, 2
-H), 3.82-3.72 (m, 1H, 5-H), 3.67 (t, J=10.2 Hz, 1H, 2-H), 2.87-2.71 (m, 2H,
-SCH.sub.2CH.sub.3), 2.61 (dddd, J=20.3, 12.9, 7.4, 5.4 Hz, 2H,
-SCH.sub.2CH.sub.3), 2.18 (s, 5H, 2CH.sub.3CO), 2.05 (s, 6H, 2CH.sub.3CO), 1.34 (t, J=7.4 Hz, 2H,
-SCH.sub.2CH.sub.3), 1.31 (t, J=6.4 Hz, 3H, 1-SCH.sub.2CH.sub.3), 1.21 (d, J=6.5 Hz, 2H, 6-CH.sub.3), 1.16 (d, J=6.5 Hz, 3H, 6-CH.sub.3).
Embodiment 13
Synthesis of 2-azido-3,4-di-O-acetyl-2-deoxy-L-fucopyranosyl 1-(N-phenyl)-2,2,2-trifluoroacetimidate (13*)
[0147] A reaction equation is as shown in
[0148] 2-azido-3,4-di-O-acetyl-2-deoxy-L-fucopyra nose (C. L. Pereira et al., Angew. Chem. Int. Ed. 2015, 54, 10016-10019) (70 mg, 0.256 mmol) was dissolved in anhydrous dichloromethane (2.6 mL), and potassium carbonate (71 mg, 0.512 mmol) and N-phenyl trifluoroacetimidoyl chloride (77 L, 0.512 mmol) were added, and stirred at room temperature overnight. After the completion of the reaction, solids were removed by filtration, dichloromethane was used for washing, and the crude product obtained by concentration was purified by silica gel column chromatography (n-hexane:ethyl acetate, 10:1, v/v) to obtain the product 13* (96.7 mg, 0.218 mmol, 85%). .sup.1H NMR (400 MHz, CDCl.sub.3) =7.40-6.75 (m, 5H, Ph), 5.58 (m, 1H, 1-H), 5.20 (m, 1H, 4-H), 4.84 (m, 1H, 3-H), 3.90 (t, J=9.3 Hz, 1H, 2-H), 3.74 (m, 1H, 5-H), 2.20 (s, 3H, CH.sub.3CO), 2.07 (s, 3H, CH.sub.3CO), 1.21 (d, J=6.4 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =170.3, 169.7, 143.0, 128.8, 124.6, 119.2, 95.7, 71.5, 70.4, 69.1, 59.9, 20.6, 15.9.
Embodiment 14
Synthesis of phenyl 2-azido-3-O-acetyl-4-O-benzyl-2-deoxy-1-seleno--D-glucopyranoside (14*)
[0149] A reaction equation is as shown in
[0150] Phenyl 2-azido-3-O-acetyl-4,6-O-benzylidene-2-deoxy-1-seleno--D-glucopyranoside (F. Santoyo-Gonzalez et al., Synlett, 1994, 6, 454-456) (1 g, 2.1 mmol) and toluene were subjected to azeotropy for three times and dissolved in 18 mL of anhydrous dichloromethane under argon protection. The 1 M borane tetrahydrofuran solution (12.4 mL, 12.4 mmol) was added to the reaction, and after cooling to 0 C., trimethylsilyl trifluoromethanesulfonate (190 L, 1.05 mmol) was added. The resulting reaction solution was stirred at room temperature. After the completion of the reaction was confirmed by TLC, the reaction solution was diluted with dichloromethane and extracted with the saturated sodium bicarbonate solution. The organic phase was dried with anhydrous sodium sulfate and subjected to reduced pressure distillation and concentration, and the resulting crude product was purified by silica gel column chromatography (n-hexane:ethyl acetate, 4:1, v/v) to obtain the yellow syrup-like compound 14* (148.9 mg, 0.313 mmol). [].sub.D.sup.20=+159.5 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3459, 2924, 2106, 1748, 1364, 1228, 1088, 737, 694 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.70-7.14 (m, 10H, 2Ph), 5.87 (d, J=5.4 Hz, 1H, 1-H), 5.38 (t, J=9.8 Hz, 1H, 3-H), 4.74-4.54 (m, 2H, PhCH.sub.2), 4.19 (dt, J=9.9, 2.9 Hz, 1H, 6-CHa), 3.90 (dd, J=10.3, 5.4 Hz, 1H, 2-H), 3.72 (m, 3H, 4-H, 5-H, 6-CH.sub.b), 2.04 (s, 3H, CH.sub.3CO), 1.64 (dd, J=7.7, 5.2 Hz, 1H, 6-OH); .sup.13C NMR (100 MHz, CDCl.sub.3) =169.7, 137.4, 134.9, 129.2, 128.6, 128.2, 128.1, 128.0, 127.9, 84.0, 75.4, 74.7, 74.2, 73.8, 63.0, 61.1, 20.9; HR-ESI-MS (m/z): calcd for C.sub.21H.sub.23N.sub.3O.sub.5SeNa.sup.+(M+Na.sup.+): 500.0701, found: 500.0682.
Embodiment 15
Synthesis of phenyl 2-azido-3-O-acetyl-4-O-benzyl-2-deoxy-6-O-(p-toluenesulfonyl)-1-seleno--D-glucopyranoside (15*)
[0151] A reaction equation is as shown in
[0152] Compound 14* (7.9 g, 16.6 mmol) was dissolved in anhydrous pyridine (110 mL), and after p-toluensulfonyl chloride (8.0 g, 42 mmol) was added, the reaction solution was stirred at room temperature overnight. After TLC showed that the starting material had been completely converted, the reaction solution was subjected to reduced pressure distillation and concentrated, diluted with dichloromethane, and then extracted with the saturated sodium bicarbonate solution and water. After dehydration with anhydrous sodium sulfate and reduced pressure distillation to remove the solvent, the resulting crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate, 5:1, v/v) to obtain the yellow syrup-like product 15* (9.06 g, 14.37 mmol, 87%). [].sub.D.sup.20=+123.2 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 2107, 1749, 1363, 1214, 1176, 1094, 977, 939, 815, 738, 681 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.83-7.14 (m, 15H, 3Ph), 5.76 (d, J=5.4 Hz, 1H, 1-H), 5.38-5.22 (m, 1H, 3-H), 4.53 (s, 2H, PhCH.sub.2), 4.38-4.21 (m, 2H, 5-H, 6-CHa), 4.09-3.97 (m, 1H, 6-CH.sub.b), 3.85 (dd, J=10.3, 5.4 Hz, 1H, 2-H), 3.65 (t, J=9.1 Hz, 1H, 4-H), 2.42 (s, 3H, PhCH.sub.3), 2.05 (s, 3H, CH.sub.3CO); .sup.13C NMR (100 MHz, CDCl.sub.3) =169.5, 145.0, 137.0, 134.5, 132.6, 129.8, 129.2, 128.6, 128.2, 128.12, 128.05, 128.0, 127.9, 84.1, 75.3, 74.9, 74.1, 71.3, 67.8, 62.7, 21.7, 20.8; HR-ESI-MS (m/z): calcd for C.sub.28H.sub.29N.sub.3O.sub.7SSeNa.sup.+(M+Na.sup.+): 654.0789, found: 654.0795.
Embodiment 16
Synthesis of phenyl 2-azido-3-O-acetyl-4-O-benzyl-2,6-dideoxy-6-iodo-1-seleno--D-glucopyranoside (16*)
[0153] A reaction equation is as shown in
[0154] Compound 15* (400 mg, 0.64 mmol) and sodium iodide (480 mg, 3.2 mmol) were added to n-butanone (8 mL) and refluxed at 80 C. for 6 hours. After cooling to room temperature, ethyl acetate was added, the mixture was extracted with the 1 M sodium thiosulfate solution and water, and the resulting organic phase was dried with anhydrous sodium sulfate. The crude product obtained by solvent removal by reduced pressure distillation was purified by silica gel column chromatography (n-hexane:ethyl acetate, 10:1, v/v) to obtain the yellow syrup-like product 16* (340 mg, 0.58 mmol, 91%). [].sub.D.sup.20=+170.3 (c=1.20, CHCl.sub.3); IR v.sub.max (film) 2928, 2108, 1746, 1367, 1226, 1091, 1044, 739, 694 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.40 (m, 10H, 2Ph), 5.89 (d, J=5.3 Hz, 1H, 1-H), 5.41 (t, J=9.7 Hz, 1H, 3-H), 4.71 (q, J=11.0 Hz, 2H, PhCH.sub.2), 3.94 (dd, J=10.3, 5.4 Hz, 1H, 2-H), 3.87 (dt, J=9.1, 3.2 Hz, 1H, 5-H), 3.59 (t, J=9.2 Hz, 1H, 4-H), 3.48 (dd, J=11.1, 3.9 Hz, 1H, 6-CHa), 3.32 (dd, J=11.1, 2.8 Hz, 1H, 6-CH.sub.b), 2.06 (s, 3H, CH.sub.3CO); .sup.13C NMR (100 MHz, CDCl.sub.3) =169.6, 137.2, 134.6, 129.3, 128.7, 128.2, 128.1, 128.0, 127.9, 84.0, 79.9, 75.1, 73.8, 71.3, 62.9, 20.9, 7.3; HR-ESI-MS (m/z): calcd for C.sub.21H.sub.22IN.sub.3O.sub.4SeNa.sup.+(M+Na.sup.+): 609.9718, found: 609.9698.
Embodiment 17
Synthesis of phenyl 2-azido-3-O-acetyl-4-O-benzyl-2-deoxy-1-seleno--D-quinovopyranoside (17*)
[0155] A reaction equation is as shown in
[0156] Compound 16* (150 mg, 0.26 mmol) was dissolved in anhydrous DMF (3.5 mL), and after the addition of sodium cyanoborohydride (82 mg, 1.30 mmol), the reaction solution was heated to 95 C. and stirred overnight. After the reaction was completed, the reaction solution was cooled to room temperature, poured into water and extracted twice with ethyl acetate. An organic phase was washed with water and then dried with anhydrous sodium sulfate, and a crude product obtained by concentration was purified by silica gel column chromatography (toluene) to obtain the white solid-like product 17* (84.4 mg, 0.18 mmol, 71%). [].sub.D.sup.20=+201.3 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 2925, 2108, 1752, 1367, 1223, 1083, 735, 692 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.63-7.22 (m, 10H, 2Ph), 5.81 (d, J=5.4 Hz, 1H, 1-H), 5.34 (dd, J=10.3, 9.1 Hz, 1H, 3-H), 4.71-4.51 (m, 2H, PhCH.sub.2), 4.26 (dq, J=9.6, 6.2 Hz, 1H, 5-H), 3.91 (dd, J=10.3, 5.4 Hz, 1H, 2-H), 3.26 (t, J=9.4 Hz, 1H, 4-H), 2.04 (s, 3H, CH.sub.3CO), 1.26 (d, J=6.2 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =169.6, 137.5, 134.7, 129.1, 128.5, 128.4, 128.03, 127.97, 127.9, 84.1, 81.8, 74.8, 74.1, 69.9, 63.3, 20.9, 17.6; HR-ESI-MS (m/z): calcd for C.sub.21H.sub.23N.sub.3O.sub.4SeNa.sup.+(M+Na.sup.+): 484.0751, found: 484.0755.
Embodiment 18
Synthesis of 2-azido-3-O-acetyl-4-O-benzyl-2-deoxy-D-quinovopyranose (18*)
[0157] A reaction equation is as shown in
[0158] Compound 17* (4.81 g, 10.5 mmol) was dissolved in the THF/water mixture (25 mL, 1:1, v/v), bromosuccinimide (4.45 g, 25.0 mmol) was added, and the reaction solution was stirred at room temperature for 6 hours. After the reaction was completed, the reaction solution was diluted with dichloromethane, and an organic phase was extracted with the 10% Na.sub.2S.sub.2O.sub.3/1 M NaHCO.sub.3 mixture (1:1, v/v). After the solvent was distilled off under reduced pressure, the crude product was purified by silica gel column chromatography (n-hexane:ethyl acetate, 5:1, v/v) to obtain the colorless syrup-like product 18* (3.31 g, 10.0 mmol, 99%). [].sub.D.sup.20=+33.90 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3402, 2934, 2111, 1751, 1363, 1227, 1077, 752, 700 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.40-7.18 (m, 10H, Ph), 5.57 (dd, J=10.3, 9.4 Hz, 1H, 3-H), 5.30 (d, J=3.3 Hz, 1H, 1-H), 5.06 (dd, J=10.2, 9.4 Hz, 1H, 3-H), 4.69 (d, J=8.0 Hz, 1H, 1-H), 4.66-4.54 (m, 4H, PhCH.sub.2), 4.13 (dq, J=12.5, 6.2 Hz, 1H, 5-H), 3.76 (s, 1H, 1-OH), 3.52 (dq, J=12.4, 6.2 Hz, 1H, 5-H), 3.35 (dd, J=10.3, 8.0 Hz, 1H, 2-H), 3.30-3.05 (m, 4H, 2-H, 4-H, 4-H, 1-OH), 2.05 (s, 6H, CH.sub.3CO), 1.34 (d, J=6.2 Hz, 3H, 6-CH.sub.3), 1.29 (d, J=6.3 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =170.01, 169.95, 137.6, 137.4, 128.52, 128.49, 128.0, 127.93, 127.88, 127.8, 95.8, 92.2, 82.1, 81.4, 75.0, 74.8, 73.9, 71.8, 71.6, 66.9, 65.6, 62.3, 20.9, 17.8; HR-ESI-MS (m/z): calcd for C.sub.15H.sub.19N.sub.3O.sub.5Na.sup.+(M+Na.sup.+): 344.1222, found: 344.1224.
Embodiment 19
Synthesis of 2-azido-3-O-acetyl-4-O-benzyl-2-deoxy-D-quinovopyranosyl trichloroacetimidate (19*)
[0159] A reaction equation is as shown in
[0160] Under nitrogen protection, trichloroacetonitrile (34 L, 0.34 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (0.6 L, 0.004 mmol) were added into an anhydrous dichloromethane (0.4 mL) solution of compound 18* (11 mg, 0.034 mmol). The resulting reaction solution was stirred at room temperature for 5 hours. After the reaction was completed, the solvent was distilled off under reduced pressure at low temperature, and a resulting crude product was purified by silica gel column chromatography (n-hexane:ethyl acetate, 7:3, v/v, containing 0.5% of triethylamine) to obtain the product 19* (13.6 mg, 0.029 mmol, 86%). .sup.1H NMR (400 MHz, CDCl.sub.3) =9.49 (s, 1H, NH), 7.46-7.23 (m, 5H, Ph), 6.48 (d, J=3.5 Hz, 1H, 1-H), 5.62-5.42 (m, 1H, 3-H), 4.72 (s, 2H, PhCH2), 4.04 (dq, J=12.5, 6.2 Hz, 1H, 5-H), 3.93 (dd, J=10.7, 3.5 Hz, 1H, 2-H), 3.49 (t, J=9.5 Hz, 1H, 4-H), 2.08 (s, 3H, CH.sub.3CO), 1.29 (d, J=6.2 Hz, 3H, 6-CH.sub.3).
Embodiment 20
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-azido-3-O-acetyl-4-O-benzyl-2-deoxy-D-quinovopyranoside (20*)
[0161] A reaction equation is as shown in
[0162] Under nitrogen protection, the trichloroacetimidate saccharide donor 19* (2.48 g, 5.325 mmol) and N-benzyl-N-benzyloxycarbonyl-3-aminopropan-1-ol (H. Ishida et al., Org. Biomol. Chem. 2015, 13, 7762-7771) (1.91 g, 6.390 mmol) were dissolved in an anhydrous diethyl ether/anhydrous dichloromethane mixture (130 mL, 3:1, v/v), and an activated molecular sieve (Aw-300) was added and stirred for 30 minutes. After the reaction solution was cooled to 40 C., trimethylsilyl trifluoromethanesulfonate (1.16 mL, 6.390 mmol) was added slowly and dropwise, and the reaction solution was stirred at this temperature until the reaction was completed. After the completion of the reaction, triethylamine was added dropwise to quench the reaction, and after the molecular sieve was removed by filtration, the solvent was distilled off under reduced pressure. A resulting crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate, 15:1, v/v) to obtain the product 20* (3.1 g, 5.14 mmol, 96%, =3:1). IR v.sub.max (film) 2919, 2107, 1749, 1696, 1454, 1421, 1361, 1222, 1044, 735, 697 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.44-7.13 (m, 20H, -3Ph, 6-3Ph), 5.49 (dd, J=10.6, 9.0 Hz, 1H, 3-H), 5.18 (d, J=8.7 Hz, 2.6H, -PhCH.sub.2, 6-PhCH.sub.2), 4.98 (s, 0.3H, 3-H), 4.82 (s, 1H, 1-H), 4.61 (m, 2.6H, -PhCH.sub.2, -PhCH.sub.2), 4.57-4.43 (m, 2.6H, -PhCH.sub.2, -PhCH.sub.2), 4.29 (d, J=8.2 Hz, 0.3H, 61-H), 3.98-3.52 (m, 2.4H, linker-OCH.sub.a, 5-H), 3.50-3.26 (m, 4.7H, linker-NCH.sub.2, linker-OCH.sub.b, 5-H, 2-H), 3.20 (t, J=9.3 Hz, 1.3H, 4-H, 4-H), 3.06 (d, J=10.4 Hz, 1H, 2-H), 2.05 (s, 3H, -CH.sub.3CO), 2.04 (s, 1H, -CH.sub.3CO), 1.86 (d, J=27.9 Hz, 2.7H, linker-CH.sub.2), 1.30 (d, J=6.2 Hz, 1H, 6-CH.sub.3), 1.26 (d, J=6.1 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =169.9, 169.8, 163.5, 156.7, 156.2, 137.9, 137.8, 137.7, 137.5, 136.8, 128.6, 128.54, 128.52, 128.49, 128.0, 127.93, 127.9, 127.8, 127.4, 101.6, 97.9, 91.9, 82.3, 81.6, 77.2, 74.9, 73.9, 72.0, 71.4, 67.3, 66.9, 66.0, 65.8, 64.6, 61.6, 50.8, 44.8, 43.9, 28.3, 27.9, 20.9, 17.8; HR-ESI-MS (m/z): calcd for C.sub.33H.sub.38N.sub.4O.sub.7Na.sup.+(M+Na.sup.+): 625.2638, found: 625.2629.
Embodiment 21
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-3-O-acetyl-4-O-benzyl-2-deoxy--D-quinovopyranoside (21*)
[0163] A reaction equation is as shown in
[0164] Thioacetic acid (0.42 mL) was added to an anhydrous pyridine (0.42 mL) solution of compound 20* (15.0 mg, 0.025 mmol) at 0 C., and the reaction solution was stirred at room temperature overnight. After the reaction was completed, the reaction solution and toluene were subjected to azeotropy to remove the solvent. Then the resulting crude product was purified by silica gel column chromatography (petroleum ether:acetone, 6:1, v/v) to obtain the colorless syrup-like product 21* (11.8 mg, 0.019 mmol, 76%). [].sub.D.sup.20=+53.1 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3343, 2936, 1745, 1696, 1455, 1423, 1366, 1233, 1120, 1048, 737, 700 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.48-7.01 (m, 15H, 3Ph), 6.51 (d, J=8.9 Hz, 1H, NH), 5.37-5.01 (m, 3H, 3-H, PhCH.sub.2), 4.80-4.31 (m, 5H, 1-H, 2PhCH.sub.2), 4.24 (m, 1H, 2-H), 3.85-3.51 (m, 3H, 5-H, linker-2H), 3.43-3.13 (m, 3H, 4-H, linker-2H), 1.99 (s, 3H, CH.sub.3CO), 1.96 (s, 3H, CH.sub.3CO), 1.75 (m, 2H, linker-CH.sub.2), 1.27 (d, J=6.2 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =171.1, 156.3, 137.9, 137.6, 136.6, 128.7, 128.5, 128.1, 127.9, 127.8, 127.5, 127.3, 97.2, 81.9, 75.1, 74.0, 67.4, 67.1, 64.1, 52.3, 49.9, 43.2, 27.3, 23.1, 21.0, 17.9; HR-ESI-MS (m/z): calcd for C.sub.35H.sub.42N.sub.2O.sub.8Na.sup.+(M+Na.sup.+): 641.2839, found: 641.2828.
Embodiment 22
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-2-deoxy--D-quinovopyranoside (22*)
[0165] A reaction equation is as shown in
[0166] Sodium methoxide (0.3 mg, 0.006 mmol) was added to the methanol (0.5 mL) solution of compound 21* (7 mg, 0.012 mmol), and the resulting reaction solution was stirred at room temperature. After the reaction was completed, the reaction solution was neutralized with an Amberlite IR 120 cation exchange resin, and the organic phase obtained by filtration was distilled off under reduced pressure. The resulting crude product was purified by silica gel column chromatography (petroleum ether:acetone, 4:1, v/v) to obtain the colorless syrup-like product 22* (6.7 mg, 0.0116 mmol, 97%). [].sub.D.sup.20=+27.9 (c=1.10, CHCl.sub.3); IR v.sub.max (film) 3325, 2938, 1695, 1544, 1424, 1369, 1235, 1120, 1070, 736, 699 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.51 (d, J=6.9 Hz, 1H, NH), 7.44-7.12 (m, 15H, 3Ph), 5.22-5.10 (m, 2H, PhCH.sub.2), 5.02 (d, J=11.0 Hz, 1H, PhCH.sub.a1), 4.78-4.63 (m, 2H, PhCH.sub.b1, PhCH.sub.a2), 4.50 (d, J=3.6 Hz, 1H, 1-H), 4.26 (d, J=15.9 Hz, 1H, PhCHb.sub.2), 4.21 (s, 1H, 3-OH), 4.00 (m, 2H, 2-H, linker-1H), 3.91 (t, J=9.4 Hz, 1H, 3-H), 3.67 (m, 2H, 5-H, linker-1H), 3.21-3.07 (m, 2H, 4-H, linker-1H), 3.00 (dt, J=14.2, 4.5 Hz, 1H, linker-1H), 2.12 (s, 3H, CH.sub.3CO), 1.70 (dq, J=9.6, 5.4 Hz, 2H, linker-CH.sub.2), 1.25 (d, J=6.2 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =174.1, 156.7, 138.7, 137.3, 136.5, 128.7, 128.5, 128.4, 128.3, 128.2, 127.7, 127.6, 127.3, 96.9, 84.7, 77.3, 76.0, 75.1, 67.5, 66.5, 62.9, 55.3, 49.4, 42.3, 26.9, 22.6, 17.9; HR-ESI-MS (m/z): calcd for C.sub.33H.sub.40N.sub.2O.sub.7Na.sup.+(M+Na.sup.+): 599.2733, found: 599.2750.
Embodiment 23
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 4-O-benzyl-2-acetamido-3-O-(3,4-di-O-acetyl-2-azido-2-deoxy-L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (23*)
[0167] A reaction equation is as shown in
[0168] Under nitrogen protection, the trifluoroimidate saccharide donor 13* (159 mg, 0.358 mmol) and the acceptor 22* (207 mg, 0.358 mmol) were dissolved in an anhydrous dichloromethane/anhydrous diethyl ether mixture (12 mL, 1/3, v/v). After thiophene (344 L, 4.296 mmol) and an activated molecular sieve (AW-300) were added, the resulting reaction solution was stirred at room temperature for 30 minutes. After cooling to 40 C., trimethylsilyl trifluoromethanesulfonate (6.5 L, 0.036 mmol) was added, and the reaction temperature was gradually increased to room temperature. After the completion of the reaction was confirmed by TLC, triethylamine was added dropwise at 0 C. to quench the reaction, and then the molecular sieve was removed by filtration with celite. After the organic phase was extracted with the saturated sodium bicarbonate solution, the crude product obtained by concentration was purified by silica gel column chromatography (petroleum ether:acetone, 6:1-3:1, v/v) to obtain the desired disaccharide 23* (262 mg, 0.315 mmol, 88%, :
=10:1). [
].sub.D.sup.20=178.6 (c=0.90, CHCl.sub.3); IR v.sub.max (film) 3337, 2936, 2112, 1749, 1680, 1371, 1233, 1044, 975, 751, 700 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.48-7.08 (m, 15H, 3Ph), 6.79 (d, J=9.3 Hz, 1H, NH), 5.40-5.30 (m, 2H, 1-H, 3-H), 5.14 (s, 2H, PhCH.sub.2), 5.01 (s, 1H, 4-H), 4.80-4.69 (m, 2H, PhCH.sub.2), 4.60 (m, 2H, 1-H, NCH.sub.aPh), 4.40 (m, 2H, NCH.sub.bPh, 2-H), 4.30-4.18 (m, 1H, 5-H), 3.96 (dd, J=19.8, 9.9 Hz, 1H, 3-H), 3.80 (dd, J=9.1, 6.3 Hz, 1H, 5-H), 3.70-3.62 (m, 2H, OCH.sub.aCCH.sub.aN), 3.58 (dd, J=11.4, 2.9 Hz, 1H, 2-H), 3.29 (dd, J=10.3, 4.7 Hz, 2H, OCH.sub.bCCH.sub.bN), 3.21 (t, J=9.3 Hz, 1H, 4-H), 2.11 (s, 3H, CH.sub.3CO), 2.06 (s, 3H, CH.sub.3CO), 2.03 (s, 3H, CH.sub.3CO), 1.74 (s, 2H, OCCH.sub.2CN), 1.34 (d, J=6.2 Hz, 3H, 6-CH.sub.3), 0.71 (d, J=6.4 Hz, 3H, 6-CH.sub.3).; .sup.13C NMR (100 MHz, CDCl.sub.3) =170.4, 169.8, 156.4, 137.7, 137.5, 136.5, 128.7, 128.6, 128.5, 128.1, 128.0, 127.8, 127.7, 127.5, 127.2, 97.8, 97.5, 83.6, 75.7, 75.5, 70.7, 68.4, 67.6, 67.4, 64.9, 63.9, 57.8, 53.4, 49.8, 43.0, 27.3, 23.1, 20.8, 20.7, 18.2, 15.5; HR-ESI-MS (m/z): calcd for C.sub.43H.sub.53N.sub.5O.sub.12Na.sup.+(M+Na.sup.+): 854.3588, found: 854.3582.
Embodiment 24
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-3-O-(2-amino-3,4-di-O-acetyl-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (24*)
[0169] A reaction equation is as shown in
[0170] Compound 23* (253 mg, 0.304 mmol) and triphenylphosphine (104 mg, 0.395 mmol) were dissolved in 5 mL of tetrahydrofuran, and the reaction solution was stirred at 40 C. After the complete reaction of the materials was confirmed by TLC, water (66 L, 3.65 mmol) was added to the reaction solution. The reaction was heated to reflux at 65 C. After the reaction was completed, the solvent was distilled off under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (dichloromethane:methanol, 40:1, v/v) to obtain the colorless syrup-like amino compound 24* (224 mg, 0.278 mmol, 91%). [].sub.D.sup.20=55.0 (c=1.10, CHCl.sub.3); IR v.sub.max (film) 3332, 2937, 1744, 1676, 1424, 1369, 1225, 1132, 1072, 1029, 972, 750, 699 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.49-7.12 (m, 15H, 3Ph), 7.08 (d, J=9.1 Hz, 1H, NH), 5.25-5.07 (m, 3H, 1-H, PhCH.sub.2), 4.99 (dd, J=11.0, 3.0 Hz, 1H, 3-H), 4.93 (s, 1H, 4-H), 4.83-4.66 (m, 2H, PhCH.sub.2), 4.60 (d, J=15.8 Hz, 1H, NCH.sub.aPh), 4.53 (d, J=2.9 Hz, 1H, 1-H), 4.49-4.28 (m, 2H, NCH.sub.bPh, 2-H), 4.27-4.10 (m, 1H, 5-H), 4.00 (t, J=9.7 Hz, 1H, 3-H), 3.82 (dd, J=9.0, 6.3 Hz, 1H, 5-H), 3.68 (ddd, J=15.0, 12.0, 6.1 Hz, 2H, OCH.sub.aCCH.sub.aN), 3.30 (d, J=7.7 Hz, 2H, OCH.sub.bCCH.sub.bN), 3.23-3.09 (m, 2H, 4-H, 2-H), 2.07 (s, 3H, CH.sub.3CO), 2.03 (s, 6H, 2CH.sub.3CO), 1.74 (s, 2H, OCCH.sub.2CN), 1.59 (s, 2H, NH.sub.2), 1.35 (d, J=6.1 Hz, 3H, 6-CH.sub.3), 0.61 (d, J=6.3 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =170.5, 137.7, 137.4, 128.6, 128.5, 128.0, 127.8, 127.7, 127.5, 127.2, 99.6, 97.6, 83.3, 77.2, 75.6, 75.0, 71.8, 70.7, 67.7, 67.4, 64.8, 63.6, 53.7, 49.7, 49.2, 42.9, 27.2, 23.3, 20.9, 20.6, 18.2, 15.6; HR-ESI-MS (m/z): calcd for C.sub.43H.sub.55N.sub.3O.sub.12Na.sup.+(M+Na.sup.+): 828.3683, found: 828.3733.
Embodiment 25
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-3-O-(3,4-di-O-acetyl-2-(2,2,2-trichloroethoxycarbonyl)amino-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (25*)
[0171] A reaction equation is as shown in
[0172] Compound 24* (224 mg, 0.278 mmol) was dissolved in anhydrous pyridine (7 mL), and 2,2,2-trichloroethoxycarbonyl chloride (96 L, 0.695 mmol) was added slowly and dropwise at 0 C. The reaction solution was stirred at room temperature, and after the reaction was completed, the reaction was quenched by adding 3.5 mL of methanol. The solvent was distilled off under reduced pressure, the resulting crude product was dissolved in dichloromethane and extracted with water, and the organic phase was dried with anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:acetone, 7:1, v/v) to obtain the colorless syrup-like product 25* (239 mg, 0.244 mmol, 88%). [].sub.D.sup.20=34.1 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3322, 2938, 1744, 1679, 1522, 1425, 1369, 1225, 1077, 1045, 740, 700 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.48-7.07 (m, 15H, 3Ph), 6.89 (d, J=8.9 Hz, 1H, NH), 5.45 (d, J=9.2 Hz, 1H, NH), 5.21 (d, J=2.8 Hz, 1H, 1-H), 5.13 (m, 3H, 3-H, PhCH.sub.2), 5.00 (d, J=11.9 Hz, 1H, Troc-CHa), 4.81 (m, 2H, 4-H, PhCH.sub.a), 4.59 (m, 3H, PhCH.sub.a, PhCH.sub.b, Troc-CH.sub.b), 4.45 (s, 1H, 1-H), 4.43-4.28 (m, 2H, 2-H, PhCH.sub.b), 4.27-4.07 (m, 2H, 2-H, 5-H), 3.96 (t, J=9.5 Hz, 1H, 3-H), 3.87-3.68 (m, 2H, 5-H, OCCCH.sub.aN), 3.69-3.54 (m, 1H, OCH.sub.aCCN), 3.19 (m, 3H, OCH.sub.bCCH.sub.bN, 4-H), 2.09 (s, 3H, CH.sub.3CO), 1.96 (s, 6H, 2CH.sub.3CO), 1.72 (m, 2H, OCCH.sub.2CN), 1.35 (d, J=6.1 Hz, 3H, 6-CH.sub.3), 0.58 (d, J=6.3 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =171.0, 170.5, 156.4, 154.9, 137.5, 137.4, 136.4, 128.7, 128.53, 128.46, 128.1, 128.0, 127.7, 127.4, 127.2, 97.7, 97.2, 83.3, 77.2, 75.8, 74.7, 74.5, 70.5, 68.9, 67.8, 67.4, 64.9, 63.5, 53.3, 49.7, 49.4, 42.8, 27.1, 23.0, 20.73, 20.65, 18.2, 15.4; HR-ESI-MS (m/z): calcd for C.sub.46H.sub.56Cl.sub.3N.sub.3O.sub.14Na.sup.+(M+Na.sup.+): 1002.2726, found: 1002.2787.
Embodiment 26
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-3-O-(2-(2,2,2-trichloroethoxycarbonyl)amino-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (26*)
[0173] A reaction equation is as shown in
[0174] Sodium methoxide (6.6 mg, 0.122 mmol) was added to the methanol (8 mL) solution of compound 25* (239 mg, 0.244 mmol), and stirred at room temperature until the reaction was completed. The reaction solution was neutralized with an Amberlite IR 120 cation exchange resin, and the resulting crude product was purified by silica gel column chromatography (petroleum ether:acetone, 4:1, v/v) to obtain the white solid-like product 26* (181 mg, 0.202 mmol, 83%). [].sub.D.sup.20=+6.4 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3331, 2934, 1679, 1533, 1456, 1225, 1042, 818, 737, 699 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.40-7.14 (m, 15H, 3Ph), 7.12 (d, J=9.1 Hz, 1H, NH), 6.19 (d, J=6.7 Hz, 1H, NH), 5.13 (m, 3H, 1-H, PhCH.sub.2), 4.77 (m, 3H, Troc-CH.sub.2, PhCH.sub.a), 4.60 (m, 2H, PhCH.sub.a, PhCH.sub.b), 4.44 (s, 1H, 1-H), 4.35 (m, 2H, 2-H, PhCH.sub.b), 4.15-4.01 (m, 2H, 5-H, 3-OH), 3.96 (m, 2H, 3-H, 2-H), 3.83 (m, 3H, OCCCH.sub.aN, 5-H, 3-H), 3.69-3.55 (m, 1H, OCH.sub.aCCN), 3.36 (s, 1H, 4-H), 3.33-3.05 (m, 3H, OCH.sub.bCCH.sub.bN, 4-H), 2.60 (s, 1H, 4-OH), 2.00 (s, 3H, CH.sub.3CO), 1.72 (m, 2H, OCCH.sub.2CN), 1.32 (d, J=6.1 Hz, 3H, 6-CH.sub.3), 0.82 (d, J=6.4 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =172.0, 157.5, 156.5, 137.7, 137.3, 136.4, 128.7, 128.5, 128.4, 128.1, 127.8, 127.7, 127.5, 127.2, 127.1, 97.7, 96.9, 95.5, 83.2, 77.2, 75.4, 74.8, 74.7, 71.5, 71.0, 67.8, 67.4, 65.6, 63.3, 53.6, 51.8, 49.6, 42.6, 27.0, 23.0, 18.1, 15.7; HR-ESI-MS (m/z): calcd for C.sub.42H.sub.52Cl.sub.3N.sub.3O.sub.12Na.sup.+(M+Na.sup.+): 918.2514, found: 918.2558.
Embodiment 27
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-3-O-(3-O-(2-naphthyl)methyl-2-(2,2,2-trichloroethoxycarbonyl)amino-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (27*)
[0175] A reaction equation is as shown in
[0176] Compound 26* (200 mg, 0.223 mmol) and toluene were subjected to azeotropy to remove water and then were vacuumized for half an hour. Under the protection of nitrogen, the mixture was dissolved in 3.2 mL of anhydrous toluene, and dibutyltin oxide (83 mg, 0.335 mmol) and an activated 4 molecular sieve were added. The reaction solution was heated to reflux for 1 hour and cooled to room temperature, naphthylmethylene bromine (148 mg, 0.669 mmol) and tetrabutylammonium bromide (108 mg, 0.335 mmol) were added, and the reaction solution was heated to 60 C. and stirred for 3 hours. After the reaction was completed, the reaction solution was filtered and concentrated, and the resulting crude product was purified by silica gel column chromatography (petroleum ether:acetone, 7:1, v/v) to obtain the white solid-like product 27* (212 mg, 0.204 mmol, 92%). [].sub.D.sup.20=14.2 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3013, 2932, 1735, 1670, 1515, 1454, 1215, 1091, 1053 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.93-6.95 (m, 22H, Ar-22H), 6.88 (d, J=9.5 Hz, 1H, 2-NH), 5.43 (d, J=9.7 Hz, 1H, 2-NH), 5.27-5.09 (m, 3H, 1-H, CH.sub.2), 5.05 (d, J=12.1 Hz, 1H, CH.sub.2-1H), 4.86-4.69 (m, 2H, CH.sub.2), 4.65-4.51 (m, 3H, NCH.sub.aPh, CH.sub.2), 4.45 (d, J=3.5 Hz, 1H, 1-H), 4.34 (m, 3H, NCH.sub.bPh, CH.sub.2-1H, 2-H), 4.20 (ddd, J=10.2, 10.2, 3.6 Hz, 1H, 2-H), 4.01-3.84 (m, 2H, 5-H, 3-H), 3.83-3.70 (m, 2H, 5-H, linker-1H), 3.69-3.60 (m, 1H, linker-1H), 3.57 (dd, J=10.7, 3.0 Hz, 1H, 3-H), 3.43 (s, 1H, 4-H), 3.26 (d, J=9.1 Hz, 1H, linker-1H), 3.20-3.10 (m, 1H, linker-1H), 3.04 (t, J=9.2 Hz, 1H, 4-H), 2.38 (s, 1H, 4-OH), 1.91 (s, 3H, CH.sub.3CO), 1.71 (t, J=6.2 Hz, 2H, linker-CH.sub.2), 1.27 (d, J=6.9 Hz, 3H, 6-CH.sub.3), 0.85 (d, J=6.5 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =171.2, 156.4, 155.0, 137.6, 137.4, 136.4, 135.1, 133.1, 128.7, 128.5, 128.3, 128.1, 127.9, 127.8, 127.7, 127.5, 127.2, 127.0, 126.7, 126.4, 126.2, 125.7, 97.9, 97.7, 95.8, 83.4, 77.2, 76.1, 75.2, 75.0, 74.6, 71.8, 68.8, 67.7, 67.4, 65.7, 63.5, 53.4, 50.1, 49.7, 42.7, 27.1, 23.1, 18.1, 15.8; HR-ESI-MS (m/z): calcd for C.sub.53H.sub.60Cl.sub.3N.sub.3O.sub.12Na.sup.+(M+Na.sup.+): 1058.3140, found: 1058.3133.
Embodiment 28
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-3-O-(4-O-[benzyl 3-azido-4-O-benzyl-2,3-dideoxy-2-trichloroacetamido-6-D-glucopyranosyl uronate]-3-O-(2-naphthyl)methyl-2-(2,2,2-trichloroethoxycarbonyl)amino-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (28*)
[0177] A reaction equation is as shown in
[0178] Under the protection of nitrogen, the trifluoroacetimidate saccharide donor 11* (69 mg, 0.096 mmol) and the disaccharide receptor 27* (20 mg, 0.019 mmol) were dissolved in 3.2 mL of anhydrous dichloromethane, and the activated molecular sieve (Aw-300) was added and stirred at room temperature for 30 minutes. After trimethylsilyl trifluoromethanesulfonate (1.8 L, 0.01 mmol) was added at room temperature, the reaction solution was continuously stirred at room temperature. After the completion of the reaction, the reaction was quenched by the addition of 4 drops of pyridine at 0 C., and the organic phase obtained by filtration was extracted with the saturated sodium bicarbonate solution and subjected to reduced pressure distillation to remove the solvent. The crude product was purified by silica gel column chromatography (petroleum ether: acetone, 6:1, v/v) to obtain the desired white solid-like trisaccharide 28* (24.6 mg, 0.016 mmol, 82%, 0-only). [].sub.D.sup.20=22.9 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3016, 2107, 1743, 1676, 1516, 1454, 1265, 1216, 1092, 1047, 752, 699 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.91-7.07 (m, 33H, 2-NH, Ar-32H), 6.81 (d, J=9.4 Hz, 1H, 2-NH), 5.37 (d, J=9.9 Hz, 1H, 2-NH), 5.22-5.05 (m, 6H, 2CH.sub.2, 1-H, 1-H), 5.01 (d, J=12.0 Hz, 1H, CH.sub.2-1H), 4.78 (d, J=11.2 Hz, 1H, CH.sub.2-1H), 4.70-4.52 (m, 6H, 5CH.sub.2-1H, 3-H), 4.51-4.40 (m, 3H, 2CH.sub.2-1H, 1-H), 4.40-4.26 (m, 2H, CH.sub.2-1H, 2-H), 4.20 (ddd, J=10.9, 10.6, 3.5 Hz, 1H, 2-H), 4.04 (d, J=9.7 Hz, 1H, 5-H), 3.97 (t, J=6.2 Hz, 1H, 5-H), 3.88 (t, J=9.7 Hz, 1H, 3-H), 3.80 (s, 1H, 4-H), 3.78-3.67 (m, 2H, linker-1H, 5-H), 3.66-3.54 (m, 2H, linker-1H, 3-H), 3.50 (t, J=9.4 Hz, 1H, 4-H), 3.34-3.11 (m, 3H, linker-2H, 2-H), 3.06 (t, J=9.3 Hz, 1H, 4-H), 1.98 (s, 3H, CH.sub.3CO), 1.74 (s, 2H, linker-CH.sub.2), 1.30-1.18 (d, J=5.3 Hz, 3H, 6-CH.sub.3), 0.80 (d, J=6.6 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =171.2, 167.7, 162.4, 156.4, 154.6, 137.7, 137.4, 137.2, 136.4, 134.8, 134.3, 133.3, 133.2, 128.8, 128.7, 128.64, 128.62, 128.5, 128.47, 128.4, 128.1, 128.0, 127.8, 127.51, 127.46, 127.3, 127.2, 126.32, 126.27, 126.2, 100.0, 97.8, 97.6, 97.0, 95.9, 91.9, 83.7, 78.7, 77.2, 75.6, 75.2, 74.7, 74.6, 73.1, 72.5, 67.6, 67.49, 67.45, 66.8, 63.6, 61.9, 57.8, 53.3, 50.9, 49.8, 42.9, 27.2, 23.2, 18.2, 16.6; HR-ESI-MS (m/z): calcd for C.sub.75H.sub.79Cl.sub.6N.sub.7O.sub.17Na.sup.+(M+Na.sup.+): 1584.3532, found: 1584.3595.
Embodiment 29
Synthesis of N-Benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-3-O-(4-O-[benzyl 3-azido-4-O-benzyl-2,3-dideoxy-2-trichloroacetamido-6-D-glucopyranosyl uronate]-2-(2,2,2-trichloroethoxycarbonyl)amino-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (29*)
[0179] A reaction equation is as shown in
[0180] Compound 28* (1.15 g, 0.734 mmol) was dissolved in the mixture of dichloromethane (6.6 mL) and water (2.6 mL), and then 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) (246 mg, 1.101 mmol) was added. The reaction solution was stirred at room temperature for 5 hours. The organic phase obtained by separation was extracted with the 5% sodium thiosulfate solution, and the crude product obtained by concentration was purified by silica gel column chromatography (petroleum ether:acetone, 7:1-6:1, v/v) to obtain the compound 29* (0.89 g, 0.625 mmol, 85%). [].sub.D.sup.20=6.5 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3317, 2934, 2107, 1704, 1517, 1216, 1039, 826, 751, 697 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.44-7.12 (m, 26H, 5Ph, 2-NH), 6.90 (d, J=9.4 Hz, 1H, 2-NH), 5.69 (d, J=8.6 Hz, 1H, 2-NH), 5.23 (q, J=12.2 Hz, 2H, CH.sub.2), 5.18-5.03 (m, 4H, CH.sub.2, 1-H, 1-H), 4.80-4.65 (m, 3H, CH.sub.2, CH.sub.a1), 4.67-4.50 (m, 4H, CH.sub.2, CH.sub.a2, CH.sub.b1), 4.46 (d, J=3.5 Hz, 1H, 1-H), 4.41-4.29 (m, 3H, CH.sub.b2, 3-H, 2-H), 4.09 (d, J=9.6 Hz, 1H, 5-H), 4.06 (d, J=6.8 Hz, 1H, 5-H), 4.01-3.84 (m, 2H, 2-H, 3-H), 3.83-3.57 (m, 5H, 5-H, 3-H, linker-2H, 4-H), 3.47 (s, 1H, 3-OH), 3.41 (s, 1H, 4-H), 3.23 (m, 3H, linker-2H, 2-H), 3.12 (t, J=9.3 Hz, 1H, 4-H), 2.00 (s, 3H, CH.sub.3CO), 1.73 (s, 2H, linker-CH.sub.2), 1.31 (d, J=6.2 Hz, 3H, 6-CH.sub.3), 0.70 (d, J=6.4 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =171.5, 167.1, 162.0, 156.4, 155.8, 137.7, 137.4, 136.9, 136.4, 134.8, 128.71, 128.67, 128.6, 128.53, 128.47, 128.4, 128.2, 128.1, 127.9, 127.8, 127.5, 127.2, 127.1, 99.6, 97.7, 95.7, 91.8, 83.4, 83.0, 78.5, 77.3, 75.4, 75.1, 74.9, 74.7, 74.6, 69.4, 67.9, 67.5, 66.5, 63.6, 62.7, 58.1, 53.4, 52.2, 49.7, 42.8, 27.1, 23.1, 18.2, 16.1; HR-ESI-MS (m/z): calcd for C.sub.64H.sub.71Cl.sub.6N.sub.7O.sub.17Na.sup.+(M+Na.sup.+): 1444.2906, found: 1444.2916.
Embodiment 30
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-3-O-(4-O-[benzyl 3-azido-4-O-benzyl-2,3-dideoxy-2-trichloroacetamido-6-D-glucopyranosyl uronate]-3-O-acetyl-2-(2,2,2-trichloroethoxycarbonyl)amino-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (30*)
[0181] A reaction equation is as shown in
[0182] Compound 29* (828 mg, 0.582 mmol) was dissolved in the anhydrous dichloromethane/pyridine mixture (30 mL, 4:1, v/v). After cooling to 0 C., acetic anhydride (550 L, 5.821 mmol) was added dropwise. After dimethylaminopyridine (1.4 mg, 0.012 mmol) was added, the reaction solution was stirred at room temperature. After the reaction was completed, the reaction solution was extracted with the saturated sodium bicarbonate solution, and the organic phase was dried with anhydrous sodium sulfate and distilled off the solvent under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:acetone, 6:1, v/v) to obtain the white solid-like compound 30* (798 mg, 0.544 mmol, 94%). [].sub.D.sup.20=37.00 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3337, 2964, 2108, 1744, 1521, 1456, 1367, 1260, 1074, 1021, 800, 752, 697 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.42-7.11 (m, 26H, 5Ph, 2-NH), 6.78 (d, J=9.5 Hz, 1H, 2-NH), 5.43 (d, J=9.9 Hz, 1H, 2-NH), 5.23-5.07 (m, 5H, 1-H, PhCH.sub.2-4H), 5.01 (d, J=12.0 Hz, 1H, CH.sub.2-1H), 4.94 (dd, J=11.8, 2.7 Hz, 1H, 3-H), 4.89 (d, J=8.1 Hz, 1H, 1-H), 4.78 (d, J=10.8 Hz, 1H, PhCH.sub.2), 4.68 (d, J=10.7 Hz, 1H, CH.sub.2-1H), 4.64-4.48 (m, 4H, CH.sub.2-4H), 4.47-4.42 (m, 2H, 1-H, 3-H), 4.37 (m, 2H, 2-H, NCH.sub.2Ph), 4.21 (ddd, J=11.4, 9.8, 3.5 Hz, 1H, 2-H), 4.04 (t, J=6.5 Hz, 1H, 5-H), 3.92 (m, 2H, 3-H, 5-H), 3.83-3.54 (m, 5H, 5-H, 4-H, 4-H, linker-2H), 3.34-3.08 (m, 4H, 4-H, 2-H, linker-2H), 1.97 (s, 3H, CH.sub.3CO), 1.96 (s, 3H, CH.sub.3CO), 1.73 (m, 2H, linker-2H), 1.33 (d, J=6.2 Hz, 3H, 6-CH.sub.3), 0.61 (d, J=6.7 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =170.9, 167.6, 162.0, 156.4, 154.8, 137.7, 137.4, 137.0, 136.4, 134.7, 128.8, 128.7, 128.53, 128.46, 128.4, 128.0, 127.9, 127.8, 127.5, 127.3, 98.7, 97.6, 97.4, 95.8, 91.9, 83.4, 79.0, 77.2, 75.6, 75.0, 74.9, 74.5, 67.8, 67.76, 67.5, 65.7, 62.3, 58.4, 53.4, 49.8, 49.2, 42.9, 27.1, 23.1, 20.9, 18.2, 15.8; HR-ESI-MS (m/z): calcd for C.sub.66H.sub.74Cl.sub.6N.sub.7O.sub.18.sup.+(M+H.sup.+): 1464.3192, found: 1464.3281.
Embodiment 31
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-3-O-(4-O-[benzyl 3-amino-4-O-benzyl-2-dichloroacetamido-2,3-dideoxy-6-D-glucopyranosyl uronate]-3-O-acetyl-2-(2,2,2-trichloroethoxycarbonyl)amino-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (31*)
[0183] A reaction equation is as shown in
[0184] Compound 30* (745 mg, 0.509 mmol) was dissolved in pyridine (51 mL), and then water (4.76 mL, 265 mmol), triethylamine (1.06 mL, 7.632 mmol) and 1,3-propanedithiol (1 mL, 10.176 mmol) were added. The reaction solution was stirred at room temperature for 6 hours. The crude product obtained by concentration of the reaction solution mixture was purified by silica gel column chromatography (petroleum ether:acetone, 5:1-3:1, v/v) to obtain the colorless syrup-like compound 31* (460 mg, 0.327 mmol, 64%). [].sub.D.sup.20=63.1 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3322, 2940, 1746, 1682, 1526, 1454, 1363, 1242, 1074, 738, 699 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.51-7.09 (m, 25H, 5Ph), 6.81 (d, J=9.3 Hz, 1H, 2-NH), 5.90 (s, 1H, CHCl.sub.2), 5.47 (s, 1H, 2-NH), 5.30-5.05 (m, 5H, 2CH.sub.2, 1-H), 4.98 (m, 2H, 3-H, CH.sub.2-1H), 4.77 (d, J=10.9 Hz, 1H, CH.sub.2-1H), 4.71 (s, 1H, 1-H), 4.67-4.49 (m, 5H, NCH.sub.aPh, CH.sub.2, 2CH.sub.2-1H), 4.45 (d, J=4.0 Hz, 1H, 1-H), 4.41-4.30 (m, 2H, NCH.sub.bPh, 2-H), 4.29-4.16 (m, 1H, 2-H), 4.14-3.98 (m, 1H, 5-H), 3.91 (m, 2H, 5-H, 3-H), 3.78 (q, J=7.5, 7.0 Hz, 1H, 5-H), 3.83-3.56 (m, 4H, 4-H, 4-H, linker-2H), 3.47 (s, 1H, 2-H), 3.36-3.06 (m, 3H, linker-2H, 4-H), 1.96 (s, 6H, 2CH.sub.3CO), 1.73 (m, 2H, linker-CH.sub.2), 1.33 (d, J=6.1 Hz, 3H, 6-CH.sub.3), 0.63 (d, J=6.5 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =171.1, 168.3, 156.4, 137.9, 137.5, 137.4, 136.5, 134.8, 128.74, 128.68, 128.5, 128.4, 128.1, 127.9, 127.8, 127.5, 127.3, 100.2, 97.6, 95.8, 83.3, 77.2, 75.6, 74.5, 74.4, 69.9, 67.8, 67.6, 67.5, 66.5, 66.0, 63.7, 54.2, 53.4, 49.8, 49.1, 27.2, 23.1, 20.9, 18.2, 15.9; HR-ESI-MS (m/z): calcd for C.sub.66H.sub.77Cl.sub.5N.sub.5O.sub.18.sup.+(M+H.sup.+): 1404.3677, found: 1404.3643.
Embodiment 32
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-3-O-(4-O-[benzyl 4-O-benzyl-3-N(R)-3-O-benzylbutyryl-2-dichloroacetamido-2,3-dideoxy-6-D-glucopyranosyl uronate]-3-O-acetyl-2-(2,2,2-trichloroethoxycarbonyl)amino-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (32*)
[0185] A reaction equation is as shown in
[0186] (R)-3-O-benzylbutyric acid (D. Seebach et al. Helv. Chim. Acta 1988, 71, 155-167) (234 mg, 1.205 mmol) was dissolved in 23 mL of anhydrous dichloromethane, and then oxalyl chloride (0.8 mL, 9.156 mmol) was added at 0 C. After the mixture was stirred at room temperature for 4 hours, the solvent and residual reagents were distilled off under reduced pressure, and obtained (R)-3-O-benzylbutyryl chloride was vacuumized for 3 hours. The amino compound 31* (113 mg, 0.081 mmol) was dissolved in 4 mL of anhydrous dichloromethane, and after triethylamine (167 L, 1.207 mmol) and the anhydrous dichloromethane solution (4 mL) of (R)-3-O-benzylbutyryl chloride were added, the reaction solution was stirred at room temperature overnight. After the reaction was completed, the reaction was quenched by the addition of 0.3 mL of methanol at 0 C. The crude product obtained by concentration was purified by silica gel column chromatography (petroleum ether:acetone, 5:1, v/v) to obtain the colorless syrup-like compound 32* (92 mg, 0.058 mmol, 72%). [].sub.D.sup.20=47.1 (c=1.00, CHCl.sub.3); IR v.sub.max (film) 3281, 2935, 1744, 1661, 1532, 1454, 1246, 1045, 737, 698 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.54 (d, J=5.1 Hz, 1H, 2-NH), 7.46-6.88 (m, 30H, 6Ph), 6.76 (d, J=9.6 Hz, 1H, 2-NH), 6.64 (d, J=9.1 Hz, 1H, 3-NH), 5.78 (s, 1H, DCA-1H), 5.40 (d, J=9.9 Hz, 1H, 2-NH), 5.29-4.94 (m, 6H, 2CH.sub.2, CH.sub.2-1H, 1-H), 4.88 (d, J=11.1 Hz, 1H, 3-H), 4.70 (s, 2H, CH.sub.2), 4.65-4.50 (m, 3H, CH.sub.2-3H), 4.50-4.45 (m, 1H, 1-H), 4.44-4.27 (m, 5H, 1-H, 2-H, 3-H, CH.sub.2), 4.23 (m, 2H, 2-H, CH.sub.2-1H), 4.02-3.85 (m, 4H, 5-H, 5-H, 3-H, 2-H), 3.79 (m, 2H, butyryl-CH, 5-H), 3.66 (m, 2H, linker-2H), 3.53 (t, J=8.6 Hz, 1H, 4-H), 3.40 (s, 1H, 4-H), 3.26 (s, 2H, linker-2H), 3.14 (t, J=9.2 Hz, 1H, 4-H), 2.40-2.12 (m, 2H, butyryl-CH.sub.2), 1.96 (s, 6H, 2CH.sub.3CO), 1.72 (s, 2H, linker-2H), 1.28 (d, J=6.3 Hz, 3H, 6-CH.sub.3), 1.19 (d, J=6.2 Hz, 3H, butyryl-CH.sub.3), 0.65 (d, J=6.4 Hz, 3H, 6-CH.sub.3); 13C NMR (100 MHz, CDCl.sub.3) =171.9, 171.2, 171.0, 168.0, 164.4, 156.4, 154.5, 137.9, 137.44, 137.35, 136.5, 134.9, 128.8, 128.71, 128.65, 128.6, 128.5, 128.42, 128.38, 128.1, 128.0, 127.9, 127.8, 127.7, 127.6, 127.4, 127.3, 102.0, 97.6, 95.9, 83.3, 77.3, 75.6, 75.3, 74.5, 74.0, 71.5, 70.4, 69.8, 67.7, 67.5, 66.3, 66.0, 63.7, 54.6, 53.4, 52.5, 49.8, 49.1, 43.6, 43.0, 27.2, 23.2, 20.9, 19.4, 18.2, 15.9; HR-ESI-MS (m/z): calcd for C.sub.77H.sub.88Cl.sub.5N.sub.5O.sub.20Na.sup.+(M+Na.sup.+): 1602.4333, found: 1602.4312.
Embodiment 33
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-3-O-(4-O-[benzyl 2-acetamido-4-O-benzyl-3-N(R)-3-O-benzylbutyryl-2,3-dideoxy-6-D-glucopyranosyl uronate]-2-amino-3-O-acetyl-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (33*)
[0187] A reaction equation is as shown in
[0188] Excessive zinc powder was added to an acetic acid (10 mL) solution of trisaccharide 32* (48.3 mg, 0.031 mmol), and the reaction solution was heated to 55 C. and stirred for 4 hours. After the reaction solution was filtered by celite, the crude product obtained by concentration was purified by silica gel column chromatography (dichloromethane:methanol, 20:1, v/v) to obtain the compound 33* (33.4 mg, 0.025 mmol, 81%). [].sub.D.sup.20=73.4 (c=0.50, CHCl.sub.3); IR v.sub.max (film) 3289, 2924, 1748, 1656, 1546, 1367, 1238, 1073, 739, 698 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.45-6.99 (m, 30H, 6Ph), 5.13 (m, 5H, 2PhCH.sub.2, 1-H), 4.93 (m, 1H, 3-H), 4.72 (s, 2H, PhCH.sub.2), 4.68-4.44 (m, 4H, NCH.sub.aPh, PhCH.sub.2-1H, 1-H, 3-H), 4.42-4.28 (m, 4H, PhCH.sub.2, PhCH.sub.2-1H, NCH.sub.bPh), 4.27-4.07 (m, 2H, 2-H, 1-H), 4.01 (d, J=6.6 Hz, 1H, 5-H), 3.90 (m, 4H, butyryl-CH, 2-H, 3-H, 5-H), 3.71 (m, 5H, 5-H, 4-H, linker-2H, 4-H), 3.50-3.03 (m, 4H, 2-H, linker-2H, 4-H), 2.34 (d, J=31.3 Hz, 2H, butyryl-CH.sub.2), 2.16-1.86 (m, 6H, 2CH.sub.3CO), 1.82 (s, 3H, CH.sub.3CO), 1.77-1.62 (m, 2H, linker-CH.sub.2), 1.30 (d, J=6.2 Hz, 3H, 6-CH.sub.3), 1.19 (d, J=6.1 Hz, 3H, butyryl-CH.sub.3), 0.80-0.48 (d, J=5.6 Hz, 3H, 6-CH.sub.3); .sup.13C NMR (100 MHz, CDCl.sub.3) =172.3, 171.0, 168.2, 156.4, 138.3, 138.0, 137.5, 136.5, 134.8, 128.7, 128.6, 128.5, 128.4, 128.3, 128.2, 128.1, 127.8, 127.7, 127.5, 127.2, 97.4, 77.2, 75.7, 75.3, 74.1, 71.8, 70.4, 67.5, 67.4, 66.0, 54.2, 53.2, 49.8, 48.7, 43.5, 43.1, 29.7, 27.2, 23.3, 21.3, 19.2, 18.1, 15.9; HR-ESI-MS (m/z): calcd for C.sub.74H.sub.90N.sub.5O.sub.18.sup.+(M+H.sup.+): 1336.6281, found: 1336.6243.
Embodiment 34
Synthesis of N-benzyl-N-benzyloxycarbonyl-3-aminopropyl 2-acetamido-4-O-benzyl-3-O-(4-O-[benzyl 2-acetamido-4-O-benzyl-3-N(R)-3-O-benzylbutyryl-2,3-dideoxy-6-D-glucopyranosyl uronate]-2-acetamidino-3-O-acetyl-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (34*)
[0189] A reaction equation is as shown in
[0190] Under protection of argon, the amino compound 33* (12.5 mg, 9.35 mol) was dissolved in 2 mL of anhydrous pyridine. After cooling to 0 C., benzyl thioacetimidate hydrochloride (3.8 mg, 18.7 mol) was added. The reaction solution was stirred at 0 C. for 5 hours, and the crude product obtained by concentration was purified by silica gel column chromatography (dichloromethane:methanol, 20:1, v/v) to obtain the colorless syrup-like compound 34* (9.0 mg, 6.53 mol, 70%). [].sub.D.sup.20=72.7 (c=0.50, CHCl.sub.3); IR v.sub.max (film) 3292, 1749, 1657, 1564, 1373, 1232, 1074, 1047, 739, 698 cm.sup.1; .sup.1H NMR (400 MHz, CDCl.sub.3) =7.49 (d, J=8.7 Hz, 2H, 2-NH, 3-NH), 7.44-7.15 (m, 30H, 6Ph), 6.96 (s, 1H, 2-NH), 5.28-5.07 (m, 4H, 2PhCH.sub.2), 4.99 (m, 2H, 1-H, 3-H), 4.83 (d, J=10.8 Hz, 1H, PhCH.sub.2-1H), 4.67 (d, J=15.7 Hz, 1H, PhCH.sub.2-1H), 4.61-4.37 (m, 6H, 2PhCH.sub.2, PhCH.sub.2-1H, 1-H), 4.35-4.11 (m, 5H, PhCH.sub.2-1H, 1-H, 2-H, 2-H, 3-H), 4.06 (d, J=6.6 Hz, 1H, 5-H), 3.92 (m, 6H, linker-1H, butyryl-CH, 3-H, 2-H, 4-H, 5-H), 3.86-3.77 (m, 1H, 5-H), 3.67 (m, 2H, linker-1H, 4-H), 3.17 (m, 2H, linker-1H, 4-H), 3.06 (d, J=14.2 Hz, 1H, linker-1H), 2.59 (s, 3H, AmCH.sub.3), 2.39 (m, 2H, butyryl-CH.sub.2), 2.09 (s, 3H, CH.sub.3CO), 2.02 (s, 3H, CH.sub.3CO), 1.81 (s, 3H, CH.sub.3CO), 1.73 (m, 2H, linker-CH.sub.2), 1.36 (d, J=6.4 Hz, 3H, 6-CH.sub.3), 1.17 (d, J=5.8 Hz, 3H, butyryl-CH.sub.3), 0.50 (d, J=6.3 Hz, 3H, 6-CH.sub.3).; .sup.13C NMR (100 MHz, CDCl.sub.3) =172.9, 170.6, 168.2, 166.8, 156.6, 138.5, 137.7, 137.4, 136.3, 134.8, 128.8, 128.7, 128.65, 128.6, 128.5, 128.4, 128.3, 128.2, 128.0, 127.9, 127.71, 127.65, 127.6, 127.5, 127.3, 102.3, 97.6, 96.4, 83.2, 77.2, 76.0, 75.8, 75.1, 74.3, 71.9, 70.8, 70.4, 68.0, 67.7, 67.5, 66.2, 63.3, 54.0, 53.0, 50.1, 49.7, 43.5, 42.6, 27.0, 23.3, 21.2, 19.8, 19.5, 18.2, 15.4; HR-ESI-MS (m/z): calcd for C.sub.76H.sub.93N.sub.6O.sub.18.sup.+(M+H.sup.+): 1377.6546, found: 1377.6526.
Embodiment 35
Synthesis of 3-aminopropyl 2-acetamido-3-O-(4-O-[2-acetamido-2,3-dideoxy-3-N(R)-3-hydroxybutyryl-6-D-glucopyranosyl uronate]-2-acetamidino-3-O-acetyl-2-deoxy--L-fucopyranosyl)-2-deoxy--D-quinovopyranoside (35*)
[0191] A reaction equation is as shown in
[0192] Trisaccharide 34* (4.3 mg, 3.1 mol) was dissolved in the t-butanol/water/dichloromethane mixture (3 mL, 5:2:1, v/v/v). After the reaction system was replaced with nitrogen, the 10% palladium on carbon hydrogenation catalyst was added, and nitrogen replacement was continued for 5 minutes. After further replacing the reaction system with hydrogen for 5 minutes, the reaction solution was stirred in the hydrogen atmosphere for 24 hours, the crude product obtained by celite filtration and concentration was preliminarily purified by the C18 column (Macherey-Nagel, Dren, Germany) (eluent were water and methanol), and the product was further purified by reversed phase high performance liquid chromatography (semipreparative Thermo Scientific Hypercarb column) to obtain the white solid-like target product 35* (1.9 mg, 2.4 mol, 77%). [
].sub.D.sup.20=66.26 (c=0.10, H.sub.2O); .sup.1H NMR (700 MHz, D.sub.2O) =5.18 (d, J=3.9 Hz, 1H, 1-H), 5.10 (dd, J=11.3, 2.7 Hz, 1H, 3-H), 4.74 (d, J=3.6 Hz, 1H, 1-H), 4.61 (d, J=8.3 Hz, 1H, 1-H), 4.53 (q, J=6.7 Hz, 1H, 5-H), 4.26 (dd, J=11.1, 3.9 Hz, 1H, 2-H), 4.21-4.12 (m, 3H, butyryl-CH, 4-H, 2-H), 4.05-3.99 (m, 1H, 3-H), 3.92-3.84 (m, 1H, 2-H), 3.83-3.75 (m, 3H, 5-H, 3-H, linker-CHa), 3.73 (d, J=9.7 Hz, 1H, 5-H), 3.71-3.62 (m, 1H, 4-H), 3.55 (ddd, J=21.7, 11.5, 6.1 Hz, 1H, linker-CH.sub.b), 3.34 (t, J=9.3 Hz, 1H, 4-H), 3.13-3.03 (m, 2H, linker-CH.sub.2), 2.49 (dd, J=14.1, 7.4 Hz, 1H, butyryl-CH.sub.2), 2.42-2.36 (m, 1H, butyryl-CH.sub.2), 2.26 (s, 3H, AmCH.sub.3), 2.10 (s, 3H, CH.sub.3CO), 2.01 (s, 3H, CH.sub.3CO), 2.00-1.91 (m, 5H, linker-CH.sub.2, CH.sub.3CO), 1.31 (d, J=6.2 Hz, 3H, 6-CH.sub.3), 1.20 (d, J=6.4 Hz, 6H, 6-CH.sub.3, butyryl-CH.sub.3); .sup.13C NMR (176 MHz, D.sub.2O) =175.1, 174.8, 174.4, 173.6, 173.1, 166.1, 102.7, 96.9, 95.8, 78.9, 77.4, 75.9, 73.5, 70.4, 70.2, 67.8, 66.7, 65.0, 64.9, 54.3, 53.9, 53.4, 50.3, 45.1, 37.2, 27.1, 22.2, 21.8, 21.6, 20.3, 18.8, 16.7, 15.0; HR-ESI-MS (m/z): calcd for C.sub.33H.sub.57N.sub.6O.sub.16+(M+H.sup.+): 793.3831, found: 793.3812.