Preparation of New Flavin Derivatives
J . Org. Chem., Vol. 62, No. 11, 1997 3527
MCA] m/ z 517.2343. Anal. Calcd for C24H33N6O5P‚1.5 H2O
(543.56): C, 52.56; H, 6.60; P, 6.40. Found: C, 52.78; H, 6.64;
P, 6.41.
pyridine-THF (1:1, 10 mL) under argon in the dark, and
triethylamine (1 mL) was added. The mixture was stirred for
5 h, and then water was added. The resulting solution was
extracted with dichloromethane to remove the traces of un-
reacted starting compound 13. The aqueous layer was evapo-
rated to dryness, and the residue was chromatographed on C18
reversed-phase (C18-Sep-Pak Waters, 1 g cartridge) in water
to yield the acetylated intermediate 14. A solution of this
compound in concentrated aqueous ammonia (15 mL) was
stirred for 3 h. After evaporation, the residue was chromato-
graphed on a short column of reversed phase (C18-Sep-Pak
Waters, 1 g cartridge) in water-methanol (90:10). Precipita-
tion from diethyl ether yielded compound 15 (75 mg, 0.1 mmol,
75%).
Compound 11b: mp 192 °C; 31P NMR (81 MHz, CDCl3) δ
6.6; 1H NMR (300 MHz, MeOD) δ 8.10 (1H, br s), 7.90 (1H, s),
4.95 (2H, m), 4.15 (4H, m), 3.30 (2H, m), 2.90 (2H, m), 2.60
(3H, s), 2.48 (3H, s), 2.10 (2H, m), 1.90 (2H, m), 1.80 (4H, m),
1.20 (12H, dd); LRMS [FAB+, glycerol] m/ z 559 [M + H]+; 506
[M + H - CH2CH2CN]+, 461 [M + H - N(iPr)2]+; HRMS exact
mass calcd for C27H39N6O5P 559.2797 [M + H]+, found [FAB+,
MCA] m/ z 559.2806. Anal. Calcd for C27H39N6O5P‚1.5H2O
(567.62): C, 57.08; H, 6.87; P, 5.46. Found: C, 57.29; H, 7.02;
P, 5.52.
F la vin H-P h osp h on a tes 12a ,b. Compound 9 (a , 50 mg;
b, 57 mg, 0.160 mmol) was dissolved in dry pyridine (10 mL)
at 100 °C under argon in the dark. To the solution cooled at
30 °C were added phosphorous acid (54 mg, 0.66 mmol) and
then the arylsulfonyl chloride (0.160 mmol; a , p-toluenesulfo-
nyl chloride, 30.5 mg; b, 2,4,6-triisopropylbenzenesulfonyl
chloride, 48.5 mg). The mixture was stirred at room temper-
ature for 15 h, and the solvent was evaporated. The residue
was chromatographed on a short column of reversed phase
(C18-Sep-Pak Waters, 1g cartridge) in water to yield compound
12 (mixture of acid and its pyridinium salt 2:1; a , 54 mg, 0.130
mmol, 84%; b, 64 mg, 0.140 mmol, 90%).
Acetylated intermediate 14: 31P NMR (81 MHz, D2O) δ 0.8;
1H NMR (300 MHz, D2O) δ 7.62 (1H, s), 7.58 (1H, s), 7.35 (1H,
s), 5.82 (1H, m), 5.23 (1H, m), 4.45 (2H, br m), 4.00 (2H, m),
3.90 (1H, m), 3.86 (2H, m), 2.51 (3H, s), 2.39 (3H, s), 2.24 (1H,
m), 2.12 (4H, m), 1.74 (4H, m), 1.66 (3H, s), 1.56 (4H, br m);
LRMS [FAB+, glycerol] m/ z 690 [M + H]+.
Compound 15: mp 138 °C dec; 31P NMR (81 MHz, D2O) δ
0.9; 1H NMR (500 MHz, D2O) δ 7.74 (1H, s), 7.66 (1H, s), 7.33
(1H, d), 5.86 (1H, q), 4.45 (2H, br m), 4.03 (1H, m), 3.92 (1H,
m), 3.90 (4H, m), 2.54 (3H, s), 2.43 (3H, s), 2.20 (1H, m), 2.05
(1H, m), 1.64 (3H, s), 1.80 (2H, m), 1.71 (2H, m), 1.58 (2H, m),
1.40 (2H, m); 13C NMR-DEPT (75 MHz, D2O) δ 139.5, 133.6,
119.0, 88.7, 87.6, 74.0, 69.0, 68.0, 48.8, 42.0, 32.1, 29.5, 28.0,
27.0, 23.6, 21.6, 14.6; LRMS [FAB+, NBA] m/ z 645 [M + H]+.
Anal. Calcd for C28H34N6O10PNa‚2.5H2O (713.61): C, 47.17;
H, 5.47; N, 11.78. Found: C, 46.87; H, 5.40; N, 11.76.
F la vin -Th ym id in e P h osp h or oth ioa te 16. Compound
13 (100 mg, 0.142 mmol) was dissolved in a solution of 0.8 M
sulfur S8 in a pyridine-carbon disulfide mixture (1:1, 10 mL)
under argon in the dark. The mixture was stirred for 24 h,
and water was added. The resulting solution was extracted
with dichloromethane to remove the unreacted starting com-
pound 13. The aqueous layer was evaporated to dryness. The
residue was stirred with concentrated aqueous ammonia (15
mL) for 3 h. After evaporation, water was added, and the
resulting solution was extracted with dichloromethane. The
aqueous layer was evaporated, and the residue was chromato-
Compound 12a : mp 200 °C dec; 31P NMR (81 MHz, DMSO-
d6) δ 7.2 (J PH ) 657 Hz); 1H NMR (200 MHz, DMSO-d6) δ 11.27
(1H, s), 8.58 (d), 7.88 (1H, s), 7.79 (1H, s), 7.77 (m), 7.39 (m),
6.76 (1H, d, J HP ) 657 Hz), 4.65 (2H, t), 4.05 (2H, m), 2.49
(3H, s), 2.39 (3H, s), 2.07 (2H, m); LRMS [FAB+, NBA] m/ z
365 [M + H]+, 301 [M + H - PO2H]; UV (H2O, 50 mM, Tris
HCl, pH 7.7) λmax (ꢀ) 444 (12 600). Anal. Calcd for C15H16
-
N4O5P‚1H2O (381.30): C, 47.25; H, 4.76; N, 14.69; P, 8.12.
Found: C, 47.07; H, 4.74; N, 14.66; P, 8.01.
Compound 12b: mp 175 °C dec; 31P NMR (81 MHz, DMSO-
d6) δ 7.4 (J PH ) 656 Hz); 1H NMR (200 MHz, DMSO-d6) δ 11.28
(1H, s), 8.56 (d), 7.88 (1H, s), 7.79 (1H, s), 7.41 (m), 7.38 (m),
6.69 (1H, d, J HP ) 656 Hz), 4.56 (2H, t), 3.90 (2H, m), 2.49
(3H, s), 2.39 (3H, s), 1.65 (2H, m), 1.60 (2H, m), 1.56 (4H, m);
13C NMR (75 MHz, D2O) δ 163.3, 160.3, 153.5, 151.5, 150.1,
144.1, 142.1, 136.3, 136.2, 133.3, 133.2, 130.3, 119.0, 67.2, 48.8,
32.7, 29.3, 28.2, 27.6, 23.6, 21.8; LRMS [FAB+, glycerol] m/ z
406 [M + H]+; UV (H2O, 50 mM, Tris HCl, pH 7.7) λmax (ꢀ) 444
(11 800). Anal. Calcd for C18H22N4O5P‚1H2O (423.38): C,
51.05; H, 5.72; N, 13.24. Found: C, 51.16; H, 5.75; N, 13.28.
F la vin -Th ym id in e H-P h osp h od iester 13. To a solution
of compound 12b (0.80 g, 1.80 mmol) in dry pyridine (60 mL)
were added dropwise, under argon in the dark, a solution of
1-adamantanecarboxylic acid chloride (0.6 g, 3.0 mmol) in
pyridine (5 mL) and then a solution of 3′-acetylthymidine (0.73
g, 2.5 mmol) in pyridine (5 mL). The mixture was stirred for
24 h, and then water (30 mL) was added. The resulting
solution was extracted with dichloromethane (2 × 150 mL).
The combined organic extracts were dried over MgSO4 and
evaporated. Compound 13 was purified by chromatography
on silica gel in dichloromethane-methanol (95:5) (0.95 g, 1.35
mmol, 75%): mp 147 °C dec; 31P NMR (81 MHz, CDCl3), two
diastereoisomers A and B, δ 7.2 (d, J PH ) 704 Hz), 5.9 (d, J PH
) 708 Hz); 1H NMR (500 MHz, CDCl3), two diastereoisomers
A and B, δ 8.67 (2H, s), 8.60 (2H, s), 8.05 (2H, s), 7.50 (1H, s),
7.44 (1H, s), 7.38 (2H, s), 6.90 (1H, d, J HP ) 708 Hz), 6.89 (1H,
d, J HP ) 708 Hz), 6.35 (1H, dd, J ) 9, 5.6 Hz), 6.32 (1H, dd, J
) 9, 5.6 Hz), 5.25 (2H, m), 4.67 (4H, br m), 4.35 (4H, m), 4.16
(2H, m), 4.11 (4H, m), 2.55 (6H, s), 2.45 (6H, s), 2.43 (1H, m),
2.41 (1H, m), 2.21 (1H, m), 2.19 (1H, m), 2.08 (6H, s), 1.84
(4H, m), 1.72 (4H, m), 1.57 (6H, s), 1.54 (4H, m), 1.51 (4H, m);
H-phosphonate doublet 200 MHz δA 7.61, 6.19 (J HP ) 708 Hz),
graphed on a short column of reversed phase (C18
-Sep-Pak
Waters, 1 g cartridge) in water-methanol (95:5). Precipitation
from diethyl ether yielded pure compound 16 (68 mg, 0.094
mmol, 78%): mp 122 °C dec;
31P NMR (81 MHz, D2O), two
1
diastereoisomers A and B, δ 56.2, 56.0; H NMR (300 MHz,
D2O), two diastereomers A and B, δ 7.55 (4H, s), 7.36 (1H, s),
7.39 (1H, s), 5.95 (2H, m), 4.67 (2H, m), 4.47 (4H, br m), 4.10
(2H, m), 3.97 (4H, m), 3.93 (4H, m), 2.50 (6H, s), 2.38 (6H, s),
2.20 (2H, m), 2.10 (2H, m), 1.71 (3H, s), 1.69 (8H, m), 1.68
(3H, s), 1.56 (8H, m); 13C NMR-DEPT (75 MHz, D2O) δ 139.7,
133.4, 119.0, 88.7, 87.8, 74.3, 68.8, 67.7, 48.8, 42.0, 32.0, 29.5,
28.2, 27.4, 23.6, 21.6, 14.6; LRMS [FAB+, NBA] m/ z 661 [M
+ H]+. Anal. Calcd for C28H34N6O9SPNa‚2.5 H2O (729.67):
C, 46.10; H, 5.35; N, 11.52; P, 4.25. Found: C, 45.99; H, 5.39;
N, 11.48; P, 3.91.
F la vin -Th ym id in e Meth ylp h osp h otr iester 17. Com-
pound 13 (90 mg, 0.128 mmol) was dissolved in 10% methanol
in a mixture of N-methylimidazole-TEA-CCl4 (5:5:90, 15 mL)
under argon in the dark. The mixture was stirred for 6 h,
and dichloromethane was added. The resulting solution was
washed with water, and the aqueous layer was extracted twice
with dichloromethane. The combined organic extracts were
dried over MgSO4 and evaporated. The residue was stirred
with concentrated aqueous ammonia (15 mL) for 1 h. After
evaporation, dichloromethane was added, and the resulting
solution was washed with water and dried over MgSO4. The
residue obtained after evaporation was chromatographed on
silica gel in dichloromethane-methanol (95:5) to yield com-
δB 7.60, 6.19 (J HP ) 704 Hz); 300 MHz δA 8.08, 5.72 (J HP
)
708 Hz), δB 8.06, 5.72 (J HP ) 704 Hz); 500 MHz δA 8.67, 5.13
(J HP ) 708 Hz), δB 8.65, 5.13 (J HP ) 704 Hz); LRMS [FAB+,
NBA] m/ z 673 [M + H]+; HRMS exact mass calcd for
C30H37N6O10P 673.2387 [M + H]+, found [FAB+, MCA] m/ z
673.2368. Anal. Calcd for C30H37N6O10P‚CH3OH (704.67): C,
52.82; H, 5.86; N, 11.93. Found: C, 52.73; H, 5.90; N, 11.89.
F la vin -Th ym id in e P h osp h od iester 15. Compound 13
(100 mg, 0.142 mmol) was dissolved in 0.2 M iodine in
pound 17 (37 mg, 0.053 mmol, 42%): mp 157-158 °C dec; 31
P
NMR (81 MHz, CDCl3), two diastereoisomers A and B, δ -1.0,
1
-1.6; H NMR (300 MHz, CDCl3), two diastereomers A and
B, δ 8.60 (2H, br s), 8.05 (2H, s), 7.43 (1H, s), 7.39 (1H, s),
7.37 (2H, s), 6.32 (2H, m), 4.67 (4H, br m), 4.54 (2H, m), 4.30
(2H, m), 4.12 (4H, m), 3.80 (4H, m), 2.56 (6H, s), 2.44 (6H, s),
2.18 (4H, m), 1.93 (3H, s), 1.92 (3H, s), 1.85 (4H, m), 1.72 (4H,