I. Hayakawa et al. / Bioorg. Med. Chem. 20 (2012) 5745–5756
5753
tography on silica gel (2.5 g, CHCl3–MeOH = 100:1 ? 20:1) to give
alcohol 32a (107 mg, 89%) as a white solid: IR (CHCl3) 3342, 2975,
1655, 1558 cmꢁ1; 1H NMR (270 MHz, CDCl3) d 5.98 (br s, 1H), 4.67
(br s, 1H), 3.60 (t, J = 6.5 Hz, 2H), 3.19 (q, J = 6.5 Hz, 2H), 3.07 (q,
J = 6.5 Hz, 2H), 2.16 (t, J = 7.2 Hz, 2H), 1.65 (quin, J = 7.2 Hz, 2H),
1.57 (quin, J = 6.5 Hz, 2H), 1.50–1.41 (m, 6H), 1.44 (s, 9H), 1.37–
1.29 (m, 4H), (A signal due to one proton (OH) was not observed);
13C NMR (125 MHz, CDCl3) d 173.0, 156.1, 79.1, 62.5, 40.1, 39.1,
36.6, 32.2, 29.9, 29.4, 28.4 (3C), 26.1, 26.0, 25.3, 25.1; HRMS (ESI)
m/z 353.2423, calcd for C17H34N2NaO4 [M+Na]+ 353.2411.
(s, 3H), 3.86 (s, 3H), 3.84 (s, 3H), 3.20–3.13 (m, 2H), 2.96–2.90
(m, 2H), 2.20 (t, J = 7.3 Hz, 2H), 1.95–1.89 (m, 2H), 1.74–1.25 (m,
12H), (Signals due to two protons (NH) were not observed); 13C
NMR (67.8 MHz, CDCl3) d 175.2, 171.6, 153.8, 151.8, 151.6, 148.2,
139.1, 138.8, 137.1, 136.6, 121.2, 118.5, 117.6, 109.7, 105.3,
104.0, 102.0, 67.4, 60.1, 56.9, 56.6, 40.5, 39.2, 37.5, 36.7, 36.6,
36.0, 32.0, 29.7, 29.5, 25.5; HRMS (ESI) m/z 607.2638, calcd for
C
31H40N2NaO9 [M+Na]+ 607.2626.
4.1.27. 6-(3-(4,7-Dimethoxybenzo[d][1,3]dioxol-5-yl)-6-meth-
oxy-4-oxo-4H-chromen-7-yloxy)-N-(6-(5-((3aS,4S,6aR)-2-oxo-
hexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)hexyl)
hexanamide (36)
4.1.24. tert-Butyl 6-(6-bromohexanamido)hexylcarbamate (33)
To a stirred solution of alcohol 32a (107 mg, 324
CH2Cl2 (3.2 mL) were added NBS (69.4 mg, 390 mol) and Ph3P
(102 mg, 389 mol) at room temperature. After being stirred at
lmol) in
l
To a stirred solution of amine 34 (2.7 mg, 3.87
biotin N-hydroxysuccinimide ester 35 (2.0 mg, 5.87
DMF (0.08 mL) was added Et3N (3.0 L, 21.6 mol) at room tem-
l
mol) and (+)-
l
lmol) in
room temperature for 14 h, the mixture was diluted with water
(5 mL) and extracted with CH2Cl2 (5 mL ꢂ 3). The combined
extracts were washed with brine (10 mL), dried (Na2SO4), and con-
centrated. The residual oil was purified by column chromatogra-
phy on silica gel (5.0 g, CHCl3–MeOH = 100:1) to give bromide 33
(51.4 mg, 40%) as a white solid: IR (CHCl3) 3327, 2970, 1649,
l
l
perature. After being stirred at room temperature for 14 h, the mix-
ture was diluted with brine (0.5 mL) and extracted with CH2Cl2
(1 mL ꢂ 3). The combined extracts were dried (Na2SO4) and con-
centrated. The residual oil was purified by column chromatogra-
phy on silica gel (0.6 g, CHCl3–MeOH = 50:1 ? 20:1) and by
recycle HPLC [JAIGEL-1H-40 (600 ꢂ 20 mm) and JAIGEL-2H-40
(600 ꢂ 20 mm), flow rate 3.8 mL/min; detection, UV 265 nm; sol-
vent CHCl3] to give O7-biotinyl glaziovianin A (36) (2.7 mg, 41%)
as a white solid: IR (CHCl3) 3340, 3031, 2974, 1719, 1643, 1562,
1553, 676 cmꢁ1 1H NMR (270 MHz, CDCl3) d 5.88 (br s, 1H), 4.61
;
(br s, 1H), 3.39 (t, J = 7.0 Hz, 2H), 3.21 (q, J = 6.5 Hz, 2H), 3.09 (q,
J = 6.5 Hz, 2 H), 2.17 (t, J = 7.3 Hz, 2H), 1.85 (quin, J = 7.0 Hz, 2H),
1.65 (quin, J = 7.3 Hz, 2H), 1.52–1.29 (m, 10H), 1.43 (s, 9H); 13C
NMR (67.8 MHz, CDCl3) d 171.8, 156.2, 79.4, 67.4, 40.5, 39.2,
36.6, 32.0, 29.7, 29.5, 28.9 (3C), 27.5, 26.7, 26.0, 25.5; HRMS (ESI)
m/z 415.1569 calcd for C17H33BrN2NaO3 [M+Na]+ 415.1567.
1510, 1286, 1202 cmꢁ1 1H NMR (270 MHz, CDCl3) d 7.90 (s, 1H),
;
7.56 (s, 1H), 6.85 (s, 1H), 6.52 (s, 1H), 6.31 (br s, 1H), 6.16 (br s,
1H), 6.00 (s, 2H), 5.27 (s, 1H), 5.02 (s, 1H), 4.60–4.53 (m, 1H),
4.39–4.31 (m, 1H), 4.10 (t, J = 5.9 Hz, 2H), 3.95 (s, 3H), 3.87 (s,
3H), 3.85 (s, 3H), 3.25–3.09 (m, 5H), 2.94–2.89 (m, 2H), 2.31 (t,
J = 7.3 Hz, 2H), 2.22 (t, J = 7.3 Hz, 2H), 1.84–1.33 (m, 20H); 13C
NMR (125 MHz, CDCl3) d 178.8, 175.3, 171.4, 170.0, 153.8, 151.8,
151.5, 148.1, 139.1, 138.6, 137.0, 136.8, 121.2, 118.4, 117.5,
109.7, 105.3, 103.9, 102.1, 67.4, 60.0, 56.9, 56.7, 55.6, 53.9, 45.4,
40.7, 40.5, 39.1, 37.5, 37.0, 36.4, 35.9, 35.5, 32.3, 29.8, 29.5, 27.8,
26.7, 26.4, 25.5; HRMS (ESI) m/z 833.3385, calcd for
4.1.25. tert-Butyl 6-(6-(3-(4,7-dimethoxybenzo[d][1,3]dioxol-5-
yl)-6-methoxy-4-oxo-4H-chromen-7-yloxy)hexanamido)
hexylcarbamate (33a)
To a stirred solution of O7-demethyl analogue 22 (6.9 mg,
18.5
lmol) and bromide 33 (21.1 mg, 53.8
lmol) in CH2Cl2
(0.27 mL) was added K2CO3 (7.4 mg, 53.6
l
mol) at 0 °C. After being
stirred at reflux for 18 h, the mixture was diluted with water
(1 mL) and extracted with CH2Cl2 (1 mL ꢂ 3). The combined ex-
tracts were washed with brine (1 mL), dried (Na2SO4), and concen-
trated. The residual oil was purified by column chromatography on
silica gel (0.6 g, hexane–EtOAc = 1:4) to give coupling compound
33a (4.3 mg, 34%) as a white solid, and recovery of O7-demethyl
analogue 22 (3.2 mg, 46%) as a white solid: IR (CHCl3) 3344,
C
41H54N4NaO11S [M+Na]+ 833.3402.
4.1.28. (E)-Methyl 6-hydroxyhex-4-enoate (38)
To a stirred solution of aldehyde 37 (535 mg, 3.77 mmol) in
MeOH (15 mL) was added NaBH4 (171 mg, 4.52 mmol) at 0 °C.
After being stirred at 0 °C for 1 h, the mixture was diluted with sat-
urated aqueous NH4Cl (8 mL) and water (30 mL) and extracted
with CH2Cl2 (20 mL ꢂ 3). The combined extracts were dried
(Na2SO4) and concentrated. The residual oil was purified by col-
umn chromatography on silica gel (12 g, EtOAc–hexane = 3:1) to
give allylic alcohol 38 (380 mg, 70%) as a white solid: IR (CHCl3)
3020, 2967, 1657, 1600, 1568, 1502, 1288 cmꢁ1
;
1H NMR
(270 MHz, CDCl3) d 7.89 (s, 1H), 7.60 (s, 1H), 6.86 (s, 1H), 6.52 (s,
1H), 6.02 (s, 2H), 5.64 (br s, 1H), 4.55 (br s, 1H), 4.10 (t,
J = 7.0 Hz, 2H), 3.96 (s, 3H), 3.87 (s, 3H), 3.85 (s, 3H), 3.24 (q,
J = 6.5 Hz, 2H), 3.10 (q, J = 6.5 Hz, 2H), 2.22 (t, J = 7.3 Hz, 2H), 1.95
(quin, J = 7.0 Hz, 2H), 1.80–1.70 (m, 2H), 1.57–1.23 (m, 10H), 1.44
(s, 9H); 13C NMR (67.8 MHz, CDCl3) d 175.3, 171.6, 156.2, 153.8,
151.8, 151.6, 148.2, 139.1, 138.8, 137.1, 136.6, 121.2, 118.5,
117.6, 109.7, 105.3, 104.0, 102.0, 79.4, 67.4, 60.1, 56.9, 56.6, 40.5,
39.2, 37.5, 36.9, 36.6, 36.0, 32.0, 29.7, 29.5, 28.9 (3C), 25.5; HRMS
(ESI) m/z 707.3139, calcd for C36H48N2NaO11 [M+Na]+ 707.3150.
3340, 2986, 1739, 1670 cmꢁ1 1H NMR (270 MHz, CDCl3) d 5.71–
;
5.68 (m, 2H), 4.34 (dt, J = 5.4, 1.6 Hz, 2H), 3.68 (s, 3H), 2.38 (t,
J = 7.6 Hz, 2H), 2.18 (dt, J = 7.6, 5.1 Hz, 2H), (A signal due to one
proton (OH) was not observed); 13C NMR (125 MHz, CDCl3) d
173.1, 132.5, 129.6, 74.2, 53.0, 33.5, 29.9; HRMS (ESI) m/z
167.0698, calcd for C7H12NaO3 [M+Na]+ 167.0679.
4.1.26. N-(6-Aminohexyl)-6-(3-(4,7-dimethoxybenzo[d][1,3]
dioxol-5-yl)-6-methoxy-4-oxo-4H-chromen-7-yloxy)
hexanamide (34)
4.1.29. (E)-Methyl 6-(tert-butyldimethylsilyloxy)hex-4-enoate
(39)
To a stirred solution of allylic alcohol 38 (370 mg, 2.57 mmol) in
DMF (26 mL) were added imidazole (385 mg, 5.65 mmol) and
TBSCl (427 mg, 2.83 mmol) at room temperature. After being stir-
red at room temperature for 2 h, the mixture was diluted with
water (10 mL) and extracted with EtOAc (15 mL ꢂ 3). The com-
bined extracts were washed with brine (10 mL), dried (Na2SO4),
and concentrated. The residual oil was purified by column chroma-
tography on silica gel (17 g, EtOAc–hexane = 30:1 ? 10:1) to give
TBS ether 39 (603 mg, 91%) as a white solid: IR (CHCl3) 2978,
To a stirred solution of coupling compound 33a (4.3 mg,
6.29
67.5
l
mol) in CH2Cl2 (0.10 mL) was added TFA (0.05 mL,
lmol) at 0 °C. The mixture was stirred at room temperature
for 4 h and concentrated. The residual oil was purified by column
chromatography on silica gel (0.6 g, CHCl3–MeOH = 20:1 ? 10:1)
to give amine 34 (TFA salt; 2.7 mg, 62%) as a white solid: IR (CHCl3)
3326, 3024, 2980, 1648, 1591, 1554, 1510, 1285 cmꢁ1 1H NMR
;
(270 MHz, CDCl3) d 7.89 (s, 1H), 7.57 (s, 1H), 6.85 (s, 1H), 6.50 (s,
1H), 6.20 (br s, 1H), 6.01 (s, 2H), 4.09 (t, J = 5.9 Hz, 2H), 3.94
1743, 1661 cmꢁ1 1H NMR (500 MHz, CDCl3) d 5.72–5.64 (m, 2H),
;