Benzyl 6-O-benzyl-2-deoxy-3-O-p-methoxybenzyl-2-phthalimido-β- D -glucopyranoside (61)
2.5.3 フラグメント Aa を用いた非天然型糖鎖の合成
アルゴン雰囲気下、7 糖 75 27.5 mg(11.0 µmol)をジクロロメタン 1.5 ml に溶解させ、
Mn(OAc)3·2H2O 11.5 mg(42.9 µmol)とDDQ 3.2 mg(14.3 µmol)を順次加え、室温で13時間攪
拌した。MALDI-TOF MASS で7 糖 75 の消失を確認した後、飽和炭酸水素ナトリウム水
溶液、を 1 ml 加えて反応終了とした。反応溶液を酢酸エチルで希釈した後、有機相を分 離し、水と飽和食塩水で洗浄した。有機相を硫酸マグネシウムで乾燥後、溶媒を減圧下留 去し、得られた残渣を pTLC(トルエン:酢酸エチル = 1:3)で精製して 7 糖 76 を 24.0 mg(92%)得た。
1H NMR (400 MHz, CDCl3): δ 7.83-6.95(m, 22 H, Ar), 5.74(dd, J = 9.3, 10.7 Hz, 1 H), 5.64(dd, J
= 9.3, 10.7 Hz, 1 H), 5.39-5.04(m, 11 H), 4.95(t, J = 9.6 Hz, 1 H), 4.89(d, J = 3.6 Hz, 1 H), 4.80(d, J
= 1.5 Hz, 1 H), 4.76-4.73(m, 2 H), 4.66-4.58(m, 3 H), 4.47(br, 1 H), 4.40-4.01(m, 13 H), 3.88- 3.67(m, 11 H), 3.46-3.41(m, 4 H), 3.39-3.34(m, 1 H), 3.26-3.22(m, 1 H), 2.30(s, 3 H, CH3CO), 2.15-1.84(m, 45 H, CH3CO), 0.98(d, J = 6.6 Hz, 3 H, 6-CH3Fuc)
13C NMR (100 MHz, CDCl3): δ 170.5, 170.4, 170.2, 170.1, 170.0, 169.9, 169.8, 169.7, 169.6, 169.1, 167.7, 166.7, 138.1, 136.9, 134.5, 134.3, 133.7, 131.6, 131.1, 130.8, 128.4, 128.0, 127.5, 127.4, 127.3, 127.2, 123.9, 123.4, 98.3, 97.8, 97.3, 97.0, 96.9, 96.2, 80.2, 74.1, 74.0, 73.8, 73.3, 72.9, 72.5, 72.2, 72.1, 71.5, 70.6, 70.3, 70.0, 69.5, 69.5, 69.4, 69.3, 69.2, 69.1, 68.9, 68.7, 67.4, 65.6, 65.0, 64.1, 62.3, 62.2, 62.0, 61.8, 55.9, 54.4, 54.3, 29.8, 21.2, 21.0, 20.9, 20.8, 20.7, 20.6, 16.0
ESI-MS : 2398.70293 (C112H125O54N3Na+, [M+Na]+; calc. 2398.70251)
Benzyl O-(3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-β-D-glucopyranosyl)-(1→2)-O-(3,4,6-tri-O- acetyl-α-D-mannopyranosyl)-(1→3)-O-[(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyl)-(1→6)]- (2,4-di-O-acetyl-β-D-mannopyranosyl)-(1→4)-O-(3,6-di-O-acetyl-2-deoxy-2-phthalimido-β-D- glucopyranosyl)-(1→4)-O-[(3,4-di-O-acetyl-2-O-benzyl-α-L-fucopyranosyl)-(1→6)]-O-[(3,4-di- O-acetyl-2-O-benzyl-α-L-fucopyranosyl)-(1→3)]-2-deoxy-2-phthalimido-β-D-glucopyranoside (77).
アルゴン雰囲気下、MS 4A 200 mgと7糖76 20.5 mg(8.62 µmol)、フコシルドナー31 9.50 µmol)、DTBMP 8.00 mg(38.8 µmol)を
攪拌した。これに対し、MeOTfを7.3 µl(64.7 µmol)加えた後、反応容器を攪拌しながら液 体窒素に浸し、溶媒を凍結させた。つづいて、反応容器を冷蔵庫内(4 ˚C)に84時間静置し
た。MALDI-TOF MASSで7糖76の消失を確認した後、トリエチルアミン0.1 mlを加え
て反応終了とした。混合物を酢酸エチルで希釈した後にセライト濾過し、濾液を飽和食塩 水で洗浄した。有機相を硫酸マグネシウムで乾燥後、溶媒を減圧下留去し、得られた残渣 をゲル濾過(SX-3、トルエン)と pTLC(トルエン:酢酸エチル = 1:3)で精製して 8 糖 77 を 18.7 mg(80%)得た。
1H NMR (400 MHz, CDCl3): δ 7.85-6.87(m, 27 H, Ar), 5.79(dd, J = 8.8, 10.7 Hz, 1 H), 5.67- 5.62(m, 2 H), 5.29-5.08(m, 12 H), 4.94-4.86(m, 3 H), 4.77-4.53(m, 8 H), 4.43-3.67(m, 27 H), 3.59- 3.50(m, 3 H), 3.43-3.36(m, 2 H), 2.25(s, 3 H, CH3CO), 2.10-1.79(m, 51 H, CH3CO), 1.11(d, J = 6.6 Hz, 3 H, 6-CH3Fuc), 1.00(d, J = 6.3 Hz, 1 H, 6-CH3Fuc)
13C NMR (100 MHz, CDCl3): δ 170.7, 170.5, 170.4, 170.3, 169.9, 169.8, 169.6, 169.5, 169.4, 169.1, 169.0, 167.7, 166.9, 137.9, 137.8, 136.6, 134.3, 134.1, 133.7, 131.5, 131.3, 131.2, 131.1, 128.5, 128.2, 128.0, 127.9, 127.6, 127.4, 127.3, 127.2, 123.6, 123.4, 98.5, 98.3, 98.0, 97.1, 97.0, 96.5, 96.4, 96.3, 96.0, 77.2, 75.0, 74.1, 73.6, 72.7, 72.6, 72.2, 72.1, 72.0, 71.9, 71.8, 71.7, 71.6, 71.5, 70.7, 70.4, 70.2, 69.9, 69.4, 69.3, 69.0, 68.9, 68.8, 68.7, 68.2, 65.7, 64.9, 64.8, 64.6, 64.2, 62.6, 62.4, 61.9, 61.8, 56.5, 54.9, 54.3, 21.1, 20.9, 20.8, 20.7, 20.6, 20.5, 20.3, 16.0, 15.9
ESI-MS : 2718.82648 (C129H145O60N3Na+, [M+Na]+; calc. 2718.82849)
Benzyl O-(3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-β-D-glucopyranosyl)-(1→2)-O-(3,4,6-tri-O- acetyl-α-D-mannopyranosyl)-(1→3)-O-[(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyl)-(1→6)]- (2,4-di-O-acetyl-β-D-mannopyranosyl)-(1→4)-O-(3,6-di-O-acetyl-2-deoxy-2-phthalimido-β-D- glucopyranosyl)-(1→4)-2-deoxy-2-phthalimido-β-D-glucopyranoside (79).
フラグメントA 8をフラグメント Aa 74に置き換えて、7 糖64と同様の方法で合成し た。フラグメントAa 74 79.1 mg(43.5 µmol)から6糖79を75.7 mg(81%、2段階)得た。
精製 : pTLC(トルエン:酢酸エチル = 1:3)
1H NMR (400 MHz, CDCl3): δ 7.82-7.00(m, 22 H, Ar), 5.84(dd, J = 8.8, 10.7 Hz, 1 H), 5.64(dd, J =
9.3, 10.7 Hz, 1 H), 5.42(d, J = 8.5 Hz, 1 H), 5.39-5.28(m, 2 H), 5.23-5.06(m, 6 H), 4.99(t, J = 9.4 Hz, 1 H), 4.81(d, J = 1.5 Hz, 1 H), 4.75-4.71(m, 2 H), 4.69(br, 2 H), 4.57-4.01(m, 17 H), 3.87- 3.67(m, 8 H), 3.53-3.49(m, 2 H), 3.41-3.37(br, 1 H), 3.23(br, 2 H), 2.30(s, 3 H, CH3CO), 2.16(s, 3 H, CH3CO), 2.10-1.90(s, 33 H, CH3CO), 1.83(s, 3 H, CH3CO)
13C NMR (100 MHz, CDCl3): δ 170.6, 170.5, 170.4, 170.3, 170.2, 170.0, 169.9, 169.8, 169.7, 169.2, 169.1, 167.6, 166.9, 137.9, 137.0, 134.4, 134.2, 133.8, 131.6, 131.3, 131.1, 130.9, 128.0, 127.9, 127.4, 127.3, 127.1, 123.6, 123.4, 98.6, 98.3, 97.5, 97.1, 97.0, 96.2, 81.7, 77.2, 76.7, 74.6, 74.1, 73.9, 73.7, 73.0, 72.8, 72.2, 72.1, 70.6, 69.7, 69.4, 69.2, 69.1, 68.9, 68.8, 68.7, 68.6, 68.1, 67.5, 65.6, 65.0, 62.4, 62.3, 62.1, 61.8, 55.9, 54.5, 54.3, 21.1, 20.9, 20.8, 20.7, 20.6, 20.5, 20.4
ESI-MS : 2168.62060 (C102H111O48N3Na+, [M+Na]+; calc. 2168.62347)
Benzyl O-(3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-β-D-glucopyranosyl)-(1→2)-O-(3,4,6-tri-O- acetyl-α-D-mannopyranosyl)-(1→3)-O-[(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyl)-(1→6)]- (2,4-di-O-acetyl-β-D-mannopyranosyl)-(1→4)-O-(3,6-di-O-acetyl-2-deoxy-2-phthalimido-β-D- glucopyranosyl)-(1→4)-O-[(3,4-di-O-acetyl-2-O-benzyl-α-L-fucopyranosyl)-(1→3)]-2-deoxy-2- phthalimido-β-D-glucopyranoside (80).
7糖64を6糖79に置き換えて、8糖65と同様の方法で合成した。6糖79 27.7 mg(12.9 µmol)から7糖80を30.4 mg(96%)得た。
精製 : pTLC(トルエン:酢酸エチル = 2:5)
1H NMR (400 MHz, CDCl3): δ 7.86-6.91(m, 27 H, Ar), 5.71(dd, J = 9.0, 10.5 Hz, 1 H), 5.65(dd, J = 9.0, 10.7 Hz, 1 H), 5.41(d, J = 8.3 Hz, 1 H), 5.31-5.09(m, 10 H), 4.93(d, J = 8.3 Hz, 1 H), 4.88(d, J
= 3.4 Hz, 1 H), 4.79(dd, J = 3.4, 10.5 Hz, 1 H), 4.74(d, J = 1.2 Hz, 1 H), 4.70-4.66(m, 2 H), 4.62- 4.59(m, 2 H), 4.55-4.52(m, 3 H), 4.50-4.17(m, 9 H), 4.13-3.71(m, 13 H), 3.66-3.38(m, 7 H), 2.28(s, 3 H, CH3CO), 2.13-1.82(s, 45 H, CH3CO), 1.10(d, J = 6.3 Hz, 3 H, 6-CH3Fuc)
13C NMR (100 MHz, CDCl3): δ 170.4, 170.2, 170.1, 169.9, 169.8, 169.7, 169.5, 169.4, 169.3, 169.1, 169.0, 167.8, 166.8, 138.0, 137.8, 136.8, 134.3, 134.1, 133.7, 131.5, 131.2, 131.1, 128.2, 128.1,
74.6, 74.5, 74.1, 73.7, 72.8, 72.8, 72.3, 72.1, 72.0, 71.9, 71.8, 71.6, 70.7, 70.4, 70.2, 70.0, 69.9, 69.3, 69.2, 69.1, 68.9, 68.8, 68.7, 68.1, 65.7, 64.9, 64.6, 62.6, 62.3, 62.0, 61.8, 56.3, 55.0, 54.3, 21.1, 21.0, 20.9, 20.8, 20.7, 20.6, 20.5, 16.0
ESI-MS : 2488.74856 (C119H131O54N3Na+, [M+Na]+; calc. 2488.74946)
O-(2-Acetamido-2-deoxy-β-D-glucopyranosyl)-(1→2)-O-(α-D-mannopyranosyl)-(1→3)-O-[(α-D-
mannopyranosyl)-(1→6)]-(β-D-mannopyranosyl)-(1→4)-O-(2-acetamido-2-deoxy-β-D- glucopyranosyl)-(1→4)-O-[(α-L-fucopyranosyl)-(1→6)]-2-acetamido-2-deoxy-β-D- glucopyranoside (81).
9 糖 54 を 7 糖 75 に置き換えて、9 糖 7 と同様の方法で合成と精製を行った。7 糖 75 12.6 mg(5.05 µmol)から7糖81を4.8 mg(75%)得た。
1H NMR (400 MHz, D2O, Acetone at δ 2.22): δ 5.17(d, J = 2.7 Hz, 0.67 H, 1-HαGlc), 5.11(br, 1 H, 1-HαMan), 4.91(br, 1 H, 1-HαMan), 4.89-4.88(m, 1 H, 1-Hα1→6Fuc), 4.77(br, 1 H, 1-HβMan), 4.69-4.64(m, 1.33 H, 1-HβGlc×2), 4.54(d, J = 8.3 Hz, 1 H, 1-HβGlc), 4.24(br, 1 H, 2-HβMan), 4.18(br, 1 H, 2-HαMan), 2.09-2.03(m, 9 H, CH3CO), 1.22-1.20(m, 3 H, 6-CH3Fuc)
13C NMR (100 MHz, D2O, Acetone at δ 30.89): δ 175.0, 174.9, 174.7, 101.5, 100.9, 100.1, 99.8, 95.5, 91.0, 80.9, 80.3, 79.5, 77.0, 76.4, 74.9, 74.7, 74.1, 73.8, 73.2, 72.6, 72.4, 70.9, 70.7, 70.4, 70.0, 69.9, 69.7, 69.0, 68.8, 67.9, 67.4, 67.3, 66.5, 62.3, 61.5, 61.2, 60.5, 55.9, 55.5, 54.3, 23.0, 22.8, 22.6, 20.7, 16.0
ESI-MS : 1282.45092 (C48H81O35N3Na+, [M+Na]+; calc. 1282.45483)
O-(2-Acetamido-2-deoxy-β-D-glucopyranosyl)-(1→2)-O-(α-D-mannopyranosyl)-(1→3)-O-[(α-D-
mannopyranosyl)-(1→6)]-(β-D-mannopyranosyl)-(1→4)-O-(2-acetamido-2-deoxy-β-D- glucopyranosyl)-(1→4)-O-[(α-L-fucopyranosyl)-(1→6)]-O-[(α-L-fucopyranosyl)-(1→3)]-2- acetamido-2-deoxy-β-D-glucopyranoside (82).
9 糖 54 を 8 糖 77 に置き換えて、9 糖 7 と同様の方法で合成と精製を行った。8 糖 77 18.2 mg(6.75 µmol)から8糖82を7.0 mg(74%)得た。
1H NMR (400 MHz, D2O, Acetone at δ 2.22): δ 5.11(br, 2 H, 1-HαMan, 1-Hα1→3Fuc), 5.06(d, J = 3.4 Hz, 0.66 H, 1-HαGlc), 4.92-4.91(m, 2 H, 1-HαMan, 1-Hα1→6Fuc), 4.74(br, 1 H, 1-HβMan), 4.69-4.65(m, 1.33 H, 1-HβGlc×2), 4.54(d, J = 8.3 Hz, 1 H, 1-HβGlc), 4.25(br, 1 H, 2-HβMan), 2.06-2.02(m, 9 H, CH3CO), 1.27(d, J = 6.8 Hz, 3 H, 6-CH3Fuc), 1.22-1.20(m, 3 H, 6-CH3Fuc)
13C NMR (125 MHz, D2O, Acetone at δ 30.89): δ 175.4, 174.9, 101.1, 100.8, 100.7, 100.4, 100.3, 100.0, 95.5×2, 95.4, 91.6, 81.7×2, 81.0, 77.1, 76.5, 75.2, 75.1, 74.8, 74.3, 73.9, 73.4, 73.2, 73.0, 72.7, 72.5, 71.0, 70.9, 70.8, 70.6, 70.2, 70.1, 69.9, 69.8, 69.1, 68.8. 68.4, 68.0, 67.5×2, 67.3, 66.6, 62.4, 61.7×2, 61.6, 61.3, 56.0, 55.6, 54.6, 23.0, 22.9, 22.7, 20.7, 16.2, 16.0
ESI-MS : 1428.50948 (C54H91O39N3Na+, [M+Na]+; calc. 1428.51274)
O-(2-Acetamido-2-deoxy-β-D-glucopyranosyl)-(1→2)-O-(α-D-mannopyranosyl)-(1→3)-O-[(α-D-
mannopyranosyl)-(1→6)]-(β-D-mannopyranosyl)-(1→4)-O-(2-acetamido-2-deoxy-β-D- glucopyranosyl)-(1→4)-2-acetamido-2-deoxy-β-D-glucopyranoside (83).
9 糖 54 を 6 糖 79 に置き換えて、9 糖 7 と同様の方法で合成と精製を行った。6 糖 79 16.2 mg(7.55 µmol)から6糖83を6.2 mg(74%)得た。
1H NMR (400 MHz, D2O, Acetone at δ 2.22): δ 5.18(d, J = 2.4 Hz, 0.65 H, 1-HαGlc), 5.12(br, 1 H, 1-HαMan), 4.91(d, J = 1.5 Hz, 1 H, 1-HαMan), 4.77(br, 1 H, 1-HβMan), 4.71-4.68(m, 0.35 H, 1-HβGlc), 4.61-4.59(m, 1 H, 1-HβGlc), 4.54(d, J = 8.3 Hz, 1 H, 1-HβGlc), 4.25(br, 1 H, 2-HβMan), 4.18(m, 1 H, 2- HαMan), 2.07-2.03(m, 9 H, CH3CO)
13C NMR (125 MHz, D2O, Acetone at δ 30.89): δ 175.4×3, 175.1, 102.0, 101.1, 100.3×2, 95.5, 95.6, 91.1, 81.0, 80.3, 79.9, 77.1, 76.5, 75.3, 75.0, 74.8, 74.2, 73.9, 73.4, 73.1 72.7, 71.1, 70.9, 70.7, 70.6, 70.5×2, 70.1, 69.9, 69.1, 68.0, 67.4, 66.6, 66.5, 62.4, 61.6, 61.3, 60.6, 56.0, 55.5, 54.3, 23.0, 22.9, 22.5
ESI-MS : 1136.39830 (C42H71O31N3Na+, [M+Na]+; calc. 1136.39692)
O-(2-Acetamido-2-deoxy-β-D-glucopyranosyl)-(1→2)-O-(α-D-mannopyranosyl)-(1→3)-O-[(α-D-
mannopyranosyl)-(1→6)]-(β-D-mannopyranosyl)-(1→4)-O-(2-acetamido-2-deoxy-β-D- glucopyranosyl)-(1→4)-O-[(α-L-fucopyranosyl)-(1→3)]-2-acetamido-2-deoxy-β-D- glucopyranoside (84).
9 糖 54 を 7 糖 80 に置き換えて、9 糖 7 と同様の方法で合成と精製を行った。7 糖 80 12.4 mg(5.03 µmol)から7糖84を4.4 mg(69%)得た。
1H NMR (400 MHz, D2O, Acetone at δ 2.22): δ 5.11(br, 2 H, 1-HαMan, 1-Hα1→3Fuc), 5.07(d, J = 3.4 Hz, 0.55 H, 1-HαGlc), 4.91(br, 1 H, 1-HαMan), 4.74(br, 1 H, 1-HβMan), 4.71-4.67(m, 0.45 H, 1-HβGlc), 4.57-4.53(m, 2 H, 1-HβGlc×2), 4.25(br, 1 H, 2-HβMan), 4.18(d, J = 2.0 Hz, 1 H, 2-HαMan), 2.05-2.02(m, 9 H, CH3CO), 1.27(d, J = 6.8 Hz, 3 H, 6-CH3Fuc)
13C NMR (100 MHz, D2O, Acetone at δ 30.89): δ 175.0×2, 174.7, 100.9, 100.2, 100.1, 100.0, 91.3, 81.4, 80.8, 76.9, 76.4, 75.9, 75.1, 74.7, 74.4, 74.3, 74.1, 73.8, 73.2, 72.8, 72.6, 70.9, 70.7, 70.5, 70.4, 69.9, 69.8, 69.0, 68.3, 67.9, 67.3, 66.5, 66.4, 62.3, 61.5, 61.2, 55.9, 55.6, 54.5, 23.0, 22.8, 22.7, 20.7, 16.2
ESI-MS : 1282.45501 (C48H81O35N3Na+, [M+Na]+; calc. 1282.45483)