General deprotection procedure
2.46 (t, J = 7.2, 2H), 2.09–2.07 (m, 6H), 1.66–1.16 (m,16H). 13C
NMR (from HSQC, D2O): d = 127.2, 105.6, 103.4, 80.62, 80.31,
79.99, 78.43, 75.3, 73.74, 73.75, 72.18, 71.87, 71.55, 71.50, 71.24,
70.93, 69.37, 69.05, 64.68, 63.12, 63.11, 62.8, 57.8, 49.99, 42.48,
42.17, 41.55, 39.36, 38.4, 37.8, 32.48, 30.92, 30.29, 28.11, 27.79,
23.42. ESMS for C46H80N8O13S (calcd. [M+Na]+: 1089.2): found
[M+Na]+ 1090.4.
Compounds were dissolved in MeOH whereupon catalytic
NaOMe was added. The reaction mixture was stirred for 2 h and
afterwards neutralized with Dowex H+, filtered, and concentrated
in vacuo. The residue was stirred in a solution of 5% H2O in TFA
for 1 h. Solvents were evaporated and the product was purified by
preparative HPLC and lyophilized from H2O–MeCN.
Biotinylated GlcNAc 5
Monovalent alkyne 3
A “Click” reaction with GlcNAc derivative 7 and alkyne 3 was
performed by the general procedure. The product was isolated
by silica gel chromatography (CH2Cl2–MeOH, 19/1 → 9/1)
(52 mg, quantitative). A deprotection reaction was performed
by the general procedure. The deprotected product (33 mg,
0.055 mmol) was dissolved in dry DMF (2 mL) under N2
A solution of the spacer 224 (160 mg, 0.5 mmol) was dissolved
in CH2Cl2 whereupon pentynoic acid 1 (73.5 mg, 0.75 mmol),
iPr2EtN (0.09 mL, 0.5 mmol) and BOP (331.7 mg, 0.75 mmol)
were added. The reaction was stirred for 18 h and followed by TLC
(CH2Cl2: MeOH 9 : 1). After, the reaction mixture was taken up in
EtOAc (100 mL) and washed twice with H2O (50 mL) and brine
(50 mL). The organic phase was dried with Na2SO4, filtered and
concentrated. The crude product was purified by silica column
i
atmosphere. The solution was basified with Pr2EtN until pH 8
whereupon succinimidyl-6-(biotinamido)hexanoic acid (20.1 mg,
0.044 mmol) was added. The mixture was stirred for 72 h and
monitored by TLC (EtOAc, MeOH, H2O 6 : 3 : 1). Crude product
was concentrated and purified by HPLC to give the compound
1
chromatography to give 3 (130 mg, 65%). H NMR (300 MHz,
CDCl3) d 6.3 (s, 1H, NH), 4.9 (s, 1H, NH), 3.6 (m, 12H, CH2),
3.3 (q, J = 6.0, 2H, CH2), 3.2 (q, J = 6.0, 2H, CH2), 2.5 (m,
2H, CH2), 2.3 (t, J = 7.1, 2H), 1.9 (q, J = 3.0, 1H, CH), 1.7
(q, J = 6.0, 4H, CH2), 1.4 (s, 9H, CH3). 13CNMR (75.5 MHz,
CDCl3): d = 171.4, (C(O)NH), 156.4 (NHC(O)C (CH3)3), 83.30
(CHaC), 79.17 (CHBoc), 70.49, 70.20, 70.09, 69.81, 69.45, 69.30
(CH2O and CaCH), 38.36, 37.72 (CH2NH), 35.36 (CH2CO),
29.84, 29.08, 28.58(CH3, Boc), 5.03(CH2). ESMS for C20H36N2O6
(calcd. [M+Na]+: 423.3): found [M+Na]+ 423.5.
1
5 (35 mg, 68%). H NMR (500 MHz, DMSO): d = 7.83 (s, 1H,
NH), 7.77 (s, 1H, NH), 7.72 (bs, 2H, NH, Htriazole), 4.30 (m, 3H),
4.13 (s, 1H, H1), 3.68 (d, J = 10.9, 2H), 3.48 (m, 6H), 3.38 (m,
4H), 3.30 (t, J = 7.6, 1H), 3.07 (m, 5H), 3.05 (s, 1H), 3.00 (s, 1H),
2.82 (t, J = 6.4, 2H), 2.57 (d, J = 12, 1H), 2.40 (t, J = 7.6, 1H),
2.04 (d, J = 5.2, 3H), 1.97 (m, 1H), 1.82 (s, 2H), 1.60 (d, J = 5.7,
4H), 1.46 (m, 4H), 1.37 (t, J = 7.0, 2H), 1.34 (m, 2H), 1.31 (m,
2H). 13C NMR (from HSQC, DMSO): d = 126.0, 104.7, 80.49,
77.67, 74.23, 73.92, 73.3, 72.98, 71.58, 68.45, 64.39, 62.67, 58.92,
58.76, 49.54, 43.13, 41.57, 39.07, 38.45, 38.13, 33.13, 32.66, 32.04,
31.41, 29.38, 28.44, 26.41, 24.54. ESMS for C42H73N9O13S (calcd
[M+H]+: 944.5): found [M+H]+ 944.4.
Biotinylated monovalent galabiose 4
A “Click” reaction with galabiose derivative 6 and alkyne 3 was
performed by the general procedure. The product was isolated
by silica gel chromatography (EtOAc–MeOH, 1 : 0–8 : 2) (125 mg,
90%). The deprotection was performed by the general procedure.
The product was isolated and purified by preparative HPLC
Tetravalent alkyne dendrimer 9
1
(CH3CN–H2O), (36.3 mg, 95%). H NMR (300 MHz, D2O) d
A solution of 824 (206 mg, 0.20 mmol) was stirred in Tesser’s base25
(20 mL) for 3 h. The mixture was acidified with aqueous KHSO4
(1M) to pH 2 and concentrated in vacuo. The crude product was
taken up in EtOAc (100 mL), and DMF (10 mL) and washed twice
with H2O (50 mL) and brine (50 mL). The organic phase was dried
over Na2SO4, filtered, and concentrated to give the free carboxylic
acid (203 mg, quantitative). The acid (203 mg, 0.20 mmol) was
7.7 (s, 1H, CHtriazole), 4.8 (s, 1H), 4.71 (s, 1H), 4.7–4.6 (m, 15H),
4.5 (s, 1H), 4.4 (t, J = 6.3, 2H), 4.3 (q, J = 8.0, 2H), 3.9 (s,
2H), 3.8–3.6 (m, 5H), 3.6–3.4 (m, 15H), 3.3 (t, J = 6.0, 2H),
3.0 (t, J = 6.8, 2H), 2.9 (t, J = 6.8, 2H), 2.8 (t, J = 6.8, 2H),
2.5 (t, J = 7.1, 2H), 2.0 (t, J = 6.0, 2H), 1.8 (t, J = 6.3, 2H),
1.5 (t, J = 6.3, 2H). 13CNMR (from HSQC, D2O): d = 126.70,
105.40 (C-1), 103.32 (C-1¢), 79.70, 77.80, 75.28, 73.62, 73.17,
72.92, 71.91, 71.72, 71.48, 71.42, 71.01, 70.85, 70.64, 69.17, 68.97,
62.85, 62.22, 49.36 (CH2Ntriazole), 40.31, 38.62, 37.36, 32.30, 30.19,
28.72, 23.45. MALDI-TOF for C30H55N5O15 (calcd. [M+Na]+:
750.4): found [M+Na]+ 750.6. A solution of the intermediate
(25 mg, 0.030 mmol) was dissolved in dry DMF (2 mL) under
N2 atmosphere. The solution was basified with iPr2EtN until pH 8
whereupon succinimidyl-6-(biotinamido)hexanoic acid (20.6 mg,
0.045 mmol) was added. The mixture was stirred for 72 h and
monitored by TLC (EtOAc, MeOH, H2O 6 : 3 : 1). Crude product
was concentrated and purified by HPLC to give the compound 4
(15 mg, 47%). 1H NMR (300 MHz, D2O): d = 7.70 (s, 1H, Htriazole),
4.83 (d, J = 3.5, 1H), 4.47 (s, 1H), 4.59 (s, 1H), 4.51–4.41 (m,
1H), 4.40–4.38 (m, 1H), 4.30–4.26 (m, 3H), 3.91 (s, 2H), 3.77–3.71
(m, 5H), 3.61–3.40 (m, 15H), 3.30 (t, J = 6.6, 2H), 3.25–3.15 (m,
1H), 3.11–3.04 (m, 6H), 2.79–2.85 (m, 3H), 2.66 (d, J = 9, 1H),
i
dissolved in dry DMF (15 mL) whereupon Pr2EtN (0.04 mL,
0.2 mmol) spacer 2 (97 mg, 0.3 mmol) and BOP (134 mg, 0.3 mmol)
were added and the mixture was stirred for 3 h. The crude was
concentrated in vacuo at 60 ◦C, and pure product was obtained
after silica column chromatography (EtOAc–MeOH, 1 : 0–19 : 1)
as a yellow oil (192 mg, 72%). 1HNMR (300 MHz, DMSO) d 8.6
(m, 2H, NH), 8.4 (m, 1H, NH), 8.1 (t, J = 5.5, 4H, NH), 7.0–6.9
(m, 6H, CHar), 6.7–6.6 (m, 3H, CHar), 4.1 (m, 4H), 3.9 (m, 8H),
3.6 (m, 4H), 3.5 (m, 8H), 3.3–3.1 (m, 20H), 2.9 (q, J = 6.8, 2H),
≡
2.7 (s, 4H, CH C), 2.4–2.2 (m, 16H), 1.7 (t, J = 6.4, 2H), 1.5 (t,
J = 6.7, 2H), 1.3 (s, 9HBoc). 13CNMR (75.5 MHz, DMSO)26 d (CO)
176.1, 171.3, 171.2, 164.8, 160.8, 142.1, 141.7, (CAR)111.5, 109.5,
≡
109.3, (CH C) 89.16, (CH3Boc) 82.8, (CH)76.7, 72.1, 71.6, (CH2)
45.7, 45.5, 45.2, 44.9, 44.7, 44.4, 44.1, 43.6, 42.7, 41.8, (CH) 39.5,
35.1, 34.7, (CH2) 33.6, (CH3) 19.6. ESMS for C68H90N8O18 (calcd.
[M+Na]+: 1329.6): found [M+Na]+ 1329.7.
2428 | Org. Biomol. Chem., 2010, 8, 2425–2429
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The Royal Society of Chemistry 2010
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