6
Tetrahedron
3′-N-(2-Phenyl-2-oxoethylaminocarbamothioyl)staurosporine
3-Methyl-1-((3′-N-staurosporine)-1-carbonyl)-1H-imidazol-3-
ACCEPTED MANUSCRIPT
(10)
ium iodide (14)
To a solution of thiocarbonylimidazolium salt 12 (49.0 mg, 0.068
mmol) in DMF (2 mL) was added a HCl salt 9 (34.9 mg, 0.204
mmol) and Et3N (47.2 ꢀL, 0.34 mmol). The reaction was stirred
at r.t. for 24 h, and extracted with EtOAc (10 mL). The organic
layer was washed with 1N HCl (2×5 mL) and brine (10 mL),
dried over anhydrous Na2SO4, and concentrated in vacuo. The
residue was purified by FCC eluting with PE-EtOAc (1:1) to
provide 10 (18.0 mg, 0.028 mmol, 41% yield) as an amorphous
Compound 13 (50 mg, 0.089 mmol) was dissolved in CH3CN (10
mL) and treated with MeI (55 ꢀL, 0.89 mmol) at r.t. The mixture
was stirred at r.t. for 24 h. The solvent was removed under
vacuum and the residue was washed with 50 mL of PE-CH2Cl2
(1:1) to provide the carbamoylimidazolium iodide 14 (48 mg,
0.068 mmol, 77% yiled) as a light yellow powder; [α]D18 +100 (c
0.07, MeOH); IR (KBr): 3404, 1725, 1676, 1454, 1344, 1313,
1
1284, 1148, 1119, 746 cm-1; H NMR (600 MHz, DMSO-d6) δ
18
white powder; Rf 0.35 (1:1 PE-EtOAc); [α]D +257 (c 0.07,
9.66 (s, 1H), 9.32 (d, J = 8.0 Hz, 1H), 8.63 (s, 1H), 8.13 (brs,
1H), 8.09 (d, J = 7.7 Hz, 1H), 8.03 (d, J = 8.5 Hz, 1H), 7.89 (brs,
1H), 7.60 (d, J = 8.0 Hz, 1H), 7.53 (t, J = 7.9 Hz, 1H), 7.51 (t, J =
7.5 Hz, 1H), 7.39 (t, J = 7.4 Hz, 1H), 7.32 (t, J = 7.6 Hz, 1H),
7.08 (m, 1H), 5.01 (s, 2H), 4.72 (m, 1H), 4.44 (brs, 1H), 3.95 (s,
3H), 2.99 (m, 1H), 2.92 (s, 3H), 2.60 (s, 3H), 2.43 (s, 3H), 2.42
(m, 1H); 13C NMR (150 MHz, DMSO-d6) δ171.9, 148.4, 138.4,
138.0, 136.2, 132.6, 129.5, 125.8, 125.5, 125.4, 125.3, 123.9,
123.7, 122.7, 121.7, 121.1, 120.6, 119.6 (2×C), 115.2, 114.2,
113.1, 108.9, 94.5, 82.9, 82.0, 60.1, 52.7, 45.4, 36.4, 33.4, 28.8,
26.9; HRESI-MS m/z 575.2393 [M–I]+ (calcd for C33H31N6O4,
575.2401).
CHCl3); IR (KBr): 2923, 2853, 1674, 1515, 1455, 1399, 1344,
1
1313, 1223, 1122, 748 cm-1; H NMR (600 MHz, DMSO-d6) δ
9.29 (d, J = 8.1 Hz, 1H), 8.61 (s, 1H), 8.06 (d, J = 7.8 Hz, 1H),
8.04 (d, J = 7.7 Hz, 2H), 8.00 (d, J = 8.5 Hz, 1H), 7.74 (d, J = 8.2
Hz, 1H), 7.68 (t, J = 7.3 Hz, 1H), 7.58 (t, J = 7.6 Hz, 2H), 7.49 (t,
J = 7.6 Hz, 2H), 7.36 (t, J = 7.4 Hz, 1H), 7.30 (t, J = 7.5 Hz, 1H),
7.08 (t, J = 7.7 Hz, 1H), 5.85 (m, 1H), 5.09 (m, 2H), 5.01 (s, 2H),
4.51 (brs, 1H), 2.93 (s, 3H), 2.85 (s, 3H), 2.71 (m, 1H), 2.34 (s,
3H), 2.28 (ddd, J = 13.0, 13.0, 7.2 Hz, 1H); 13C NMR (150 MHz,
DMSO-d6) δ 195.4, 182.5, 171.9, 139.1, 136.3, 135.5, 133.4,
132.8, 129.0, 128.8 (2×C), 127.8 (2×C), 125.7, 125.4, 125.0,
124.9, 123.8, 122.6, 121.4, 120.3, 119.5, 119.4, 115.3, 114.2,
114.0, 109.1, 94.9, 82.7, 82.3, 60.4, 54.4, 51.9, 45.5, 32.9, 29.6,
27.6; HRESI-MS m/z 644.2351 [M+H]+ (calcd for C37H34N5O4S,
644.2332).
3′-N-(2-(1H-Indol-3-yl)-2-oxoethylaminocarbonyl)staurosporine
(15)
To a solution of carbamoylimidazolium salt 14 (48.0 mg, 0.068
mmol) in DMF (2 mL) was added a TFA salt 8 (55.2 mg, 0.204
mmol) and triethylamine (47.2 ꢀL, 0.34 mmol). The reaction was
stirred at r.t. for 24 h, and then was extracted with EtOAc (10
mL). The mixture was washed with 1N HCl (2×5 mL) and
saturated brine (10 mL). The organic layer was dried over
anhydrous Na2SO4, and concentrated in vacuo. The residue was
purified by FCC eluting with CH2Cl2-EtOAc (1:3) to provide 15
(36.0 mg, 0.054 mmol, 79% yield) as an amorphous white
3′-N-(5-Phenylthiazole-2-yl)staurosporine (2)
(CF3CO)2O (10 ꢀL, 0.07 mmol), Et3N (19.5 ꢀL, 0.14 mmol) and
catalytic amount of DMAP were added sequentially to a stirred
solution of 10 (9 mg, 0.014 mmol) in CH2Cl2 (2 mL) at 0°C.
Then warmed to 25°C and stirred for 2h, the reaction was
quenched by adding 2 mL of saturated aqueous NaHCO3 at 0°C
and extracted with CH2Cl2 (3 × 5 mL). The combined organic
layers were then washed with brine, dried over anhydrous
Na2SO4, and concentrated in vacuo. The residue was purified by
FCC eluting with PE-EtOAc (4:1) to provide 2 (4.0 mg, 0.0064
mmol, 46% yield) as an amorphous white powder; Rf 0.48 (2:1
18
powder; Rf 0.42 (1:3 CH2Cl2-EtOAc); [α]D +105 (c 0.07,
CHCl3); IR (KBr): 3152, 2921, 2852, 1676, 1515, 1458, 1395,
1
1313, 1122, 747 cm-1; H NMR (600 MHz, DMSO-d6) δ 12.08
(d, J = 2.5 Hz, 1H), 9.29 (d, J = 8.0 Hz, 1H), 8.61 (s, 1H), 8.49
(d, J = 3.1 Hz, 1H), 8.21 (d, J = 7.7 Hz, 1H), 8.04 (d, J = 7.9 Hz,
1H), 7.96 (d, J = 8.4 Hz, 1H), 7.70 (d, J = 8.4 Hz, 2H), 7.52 (d, J
= 8.0 Hz, 1H), 7.48 (t, J = 7.6 Hz, 2H), 7.35 (t, J = 7.4 Hz, 1H),
7.30 (t, J = 7.5 Hz, 1H), 7.24 (t, J = 7.3 Hz, 1H), 7.21 (t, J = 7.6
Hz, 1H), 7.02 (t, J = 7.6 Hz, 1H), 6.84 (t, J = 5.3 Hz, 1H), 5.00
(s, 2H), 4.85 (m, 1H), 4.50 (d, J = 5.7 Hz, 2H), 4.24 (brs, 1H),
2.84 (s, 3H), 2.75 (s, 3H), 2.63 (m, 1H), 2.30 (s, 3H), 2.22 (ddd, J
= 12.5, 12.5, 7.0 Hz, 1H); 13C NMR (150 MHz, DMSO-d6) δ
192.0, 172.2, 158.4, 139.2, 136.6, 136.5, 133.5, 132.9, 129.4,
125.9, 125.7, 125.6, 125.2 (2×C), 124.0, 123.1, 122.8, 122.0,
121.6, 121.4, 120.5, 119.7, 119.5, 115.4, 114.3 (2×C), 114.0,
112.4, 109.3, 95.1, 84.1, 82.7, 60.6, 49.1, 47.6, 45.7, 30.0, 29.7,
27.6; HRESI-MS m/z 667.2661 [M+H]+ (calcd for C39H35N6O5,
667.2669).
18
PE-EtOAc); [α]D +103 (c 0.07, CHCl3); UV (MeOH) λmax (log
ε) 235 (3.23), 283 (3.62), 326 (3.17), 364 (3.10) nm; CD (MeOH)
λmax (ꢁε) 297 (+12.5), 265 (+0.7), 239 (+7.5), 211 (–9.2) nm; IR
(KBr) 2924, 2855, 1680, 1516, 1459, 1393, 1314, 1155, 748 cm-
1
1; H and 13C NMR, see Table 1; HRESI-MS m/z 626.2235
[M+H]+ (calcd for C37H32N5O3S, 626.2226).
3′-N-(1H-Imidazol-1-yl)carbonylstaurosporine (13)
N,N′-carbonyldiimidazole (140 mg, 0.86 mmol) was added to a
solution of staurosporine (200 mg, 0.43 mmol) in THF (10 mL).
The mixture was refluxed for 12 h, and concentrated in vacuo.
The residue was purified by FCC eluting with CH2Cl2-MeOH
(20:1) to provide 13 (202 mg, 0.36 mmol, 84% yield) as a light
18
yellow solid; Rf 0.58 (20:1 CH2Cl2-MeOH); [α]D +176 (c 0.07,
CHCl3); IR (KBr): 1681, 1531, 1514, 1459, 1393, 1313, 1220,
3′-N-(5-(1H-Indol-3-yl)oxazole-2-yl)staurosporine (3)
1
749 cm-1; H NMR (500 MHz, DMSO-d6) δ 9.31 (d, J = 7.9 Hz,
1H), 8.68 (s, 1H), 8.61 (s, 1H), 8.07 (d, J = 7.8 Hz, 1H), 8.01 (d,
J = 8.5 Hz, 1H), 7.79 (brs, 1H), 7.59 (d, J = 8.2 Hz, 1H), 7.51 (t,
J = 8.2 Hz, 1H), 7.49 (t, J = 8.2 Hz, 1H),7.37 (t, J = 7.7 Hz, 1H),
7.35 (brs, 1H), 7.31 (t, J = 7.5 Hz, 1H), 7.04 (dd, J = 8.5, 5.6 Hz,
1H), 5.00 (s, 2H), 4.68 (m, 1H), 4.50 (brs, 1H), 2.96 (m, 1H),
2.89 (s, 3H), 2.61 (s, 3H), 2.40 (m, 1H), 2.39 (s, 3H); 13C NMR
(125 MHz, DMSO-d6) δ 172.0, 150.8, 138.6, 137.2, 136.3, 132.6,
129.5, 125.9, 125.8, 125.5, 125.5, 125.3, 123.9, 122.8, 121.7,
120.6, 119.7, 119.6, 119.5, 115.2, 114.3, 113.2, 109.0, 94.7, 83.2,
82.1, 60.2, 52.3, 45.5, 33.2, 28.9, 27.0; HRESI-MS m/z 561.2242
(CF3CO)2O (20 ꢀL, 0.14 mmol), Et3N (39 ꢀL, 0.28 mmol) and
catalytic amount of DMAP were added sequentially to a solution
of 15 (18.6 mg, 0.028 mmol) in CH2Cl2 (2 mL) at 0°C. The
reaction mixture was then warmed up to 25°C and stirred for 2 h.
The reaction was quenched by adding 2 mL of saturated aqueous
NaHCO3 at 0°C and extracted with CH2Cl2 (3 × 5 mL). The
combined organic layers were then washed with brine, dried over
anhydrous Na2SO4, and concentrated in vacuo. The residue was
purified by semipreparative HPLC (85% MeOH/H2O) to yield 3
(12 mg, 0.018 mmol, 64% yield, tR 10.0 min) as a light yellow
18
powder; [α]D +274 (c 0.07, CHCl3); UV(MeOH) λmax (log ε)
224 (3.26), 283 (3.63), 325 (3.17), 364 (3.09) nm; CD (MeOH)
[M+H]+ (calcd for C32H29N6O4, 561.2245).