5278 Journal of Medicinal Chemistry, 2006, Vol. 49, No. 17
Gao et al.
(t, J ) 6.4, 2H), 2.39 (t, J ) 6.4, 2H), 1.94 (m, 2H) 0.81 (s, 3H),
0.71 (s, 3H). 13C NMR (D2O, 100 MHz): δ 174.5, 174.0, 173.6,
150.1, 149.4, 145.0, 143.2, 119.5, 88.0, 83.1, 74.2, 73.8, 72.2, 69.5,
64.8, 54.7, 46.6, 40.4, 39.8, 38.4, 36.4, 35.3, 35.1, 34.7, 31.2, 23.4,
20.9, 18.3. HRMS for C30H53N10O17P3S (M + H) calcd 951.2524,
found 951.2528. Purity, 97% (method A, tR ) 3.27 min; method
B, tR ) 3.08 min).
29.1, 28.5, 20.7, 19.6. HRMS for C29H52N6O14S (M + H) calcd
741.3335, found 741.3333. Purity, 98% (method A, tR ) 3.55 min;
method B, tR ) 3.34 min).
Compound 5d. Crude 18 (80 mg, 0.10 mmol) was dissolved in
TFA/DCM/anisole (3 mL, 90/9/1 v/v) in a plastic bottle and stirred
overnight. After workup as described for 5a and evaporation of
the TFA and DCM, the residue was purified by HPLC to give 5c
1
Compound 4f. Yield: 13 mg, 79%. 1H NMR (D2O, 400 MHz,
presat): δ 8.47 (s, 1H), 8.25 (s, 1H), 6.04 (d, J ) 5.0, 1H), 4.41
(br s, 1H), 4.16-4.03 (m, 2H), 3.83 (s, 1H), 3.67 (dd, J ) 9.6, 4.4,
1H), 3.43 (dd, J ) 9.6, 4.4, 1H), 3.30 (m, 2H), 3.19-3.09 (m,
5H), 3.09-3.02 (m, 3H), 3.02-2.98 (m, 2H), 2.98-2.88 (m, 4H),
2.72 (s, 6H), 2.50 (t, J ) 6.8, 2H), 2.29 (t, J ) 6.8, 2H), 2.03-
1.95 (m, 2H), 1.80-1.71 (m, 2H), 0.76 (s, 3H), 0.65 (s, 3H). 13C
NMR (D2O, 100 MHz): δ 174.6, 174.0, 173.0, 150.0, 148.5, 147.4,
144.8, 142.5, 118.7, 118.33, 87.1, 83.5, 74.5, 74.2, 72.2, 65.3, 54.5,
46.9, 44.5, 43.0, 38.6, 36.7, 35.6. 35.1, 31.5, 25.8, 23.4, 21.6, 21.2,
18.7. HRMS for C31H57N10O17P3S (M + H) calcd 967.2827, found
967.2917. Purity, 98% (method A, tR ) 3.32 min; method B, tR )
3.11 min).
Compound 5a. In a vial, D-pantethine (15 mg, 0.03 mmol), DTT
(5 mg, 0.03 mmol), and DIPEA (1 mL, 5 mmol) were mixed in
acetone/H2O (2 mL, 1/1 v/v) and sonicated for 2 min. This solution
was transferred to the N-6′-bromoacetylneamine solution (0.06
mmol, 4 mL), and the resulting mixture was sonicated for another
2 min and then stirred for 1 h. After evaporation of the solvent
under vacuum, the residue was dissolved in 0.1% aqueous TFA
(10 mL) and washed with ethyl acetate (2 × 3 mL). The aqueous
layer was evaporated to dryness under vacuum and redissolved in
0.05% TFA water (3 mL) before purification by HPLC to give 5a
(22 mg, 65% based on D-pantethine). In general the final workup
process and purification for 5b-d are the same as described here
for 5a. 1H NMR (D2O, 400 MHz, presat): δ 5.56 (d, J ) 3.2, 1H),
3.82 (s, 1H), 3.74 (m, 3H), 3.52-3.30 (m, 10H), 3.30-3.12 (m,
6H), 2.58 (t, J ) 6.4, 2H), 2.35 (m, 3H), 1.73 (q, J ) 12.4, 1H),
0.76 (s, 3H), 0.73 (s, 3H). 13C NMR (D2O, 100 MHz): δ 174.4,
173.9, 173.7, 96.8, 79.0, 75.9, 75.0, 72.6, 71.5, 70.5, 68.8, 68.5,
53.8, 49.7, 48.5, 40.0, 39.5, 38.3, 35.4, 35.3, 34.7, 31.2, 28.3, 20.5,
19.2. HRMS for C25H48N6O11S (M + H) calcd 641.3175, found
641.3167. Purity, 95% (method A, tR ) 3.25 min; method B, tR )
3.06 min).
Compound 5b. 14 (25 mg, 0.05 mmol) and DIPEA (1 mL, 5
mmol) were mixed in acetone/H2O (2 mL, 1/1 v/v) and sonicated
for 2 min. This solution was transferred into the N-6′-bromoacetyl-
neamine solution (0.06 mmol, 4 mL). The resulting mixture was
sonicated for another 2 min and stirred for 1 h. Workup as described
for 5a was followed by HPLC to give 5b (24 mg, 63% based on
14). 1H NMR (D2O, 400 MHz, presat): δ 5.60 (br s, 1H), 3.90 (br
s, 1H), 3.70-3.85 (m, 3H), 3.70-3.60 (m, 1H), 3.52-3.39 (m,
9H), 3.39-3.19 (m, 6H), 2.60 (t, J ) 6.5, 2H), 2.38 (m, 3H), 1.78
(q, J ) 13.2, 1H), 0.85 (s, 3H), 0.78 (s, 3H). 13C NMR (D2O, 100
MHz): δ 177.4, 175.0, 173.7, 96.1, 78.2, 77.1, 75.4, 72.7, 70.8,
70.2, 69.5, 68.4, 53.8, 49.9, 48.6, 41.5, 40.6, 39.7, 36.0, 33.1, 30.4,
29.1, 28.4, 21.0, 18.6. 31P NMR (D2O, 81 MHz): δ -0.05. HRMS
for C25H49N6O14PS (M + H) calcd 721.2838, found 721.2846.
Purity, 97% (method A, tR ) 3.10 min; method B, tR ) 2.99 min).
Compound 5c. 15 (25 mg, 0.03 mmol), DTT (5 mg, 0.03 mmol),
and DIPEA (1 mL, 5 mmol) were mixed in acetone/H2O (3 mL,
1/1 v/v). The mixture was sonicated for 2 min and stirred overnight.
This solution was transferred to the N-6′-bromoacetylneamine
solution (4 mL). The resulting mixture was sonicated for 2 min
and stirred for 1 h. Workup as described for 5a was followed by
HPLC to give 5d (26 mg, 58%). 1H NMR (D2O, 400 MHz,
presat): δ 5.57 (d, J ) 3.5, 1H), 3.89 (d, J ) 10.5, 1H), 3.83 (s,
1H), 3.79-3.70 (m, 4H), 3.51 (d, J ) 9.0, 1H), 3.48 (t, J ) 4.0,
1H), 3.45 (t, J ) 4.0, 1H), 3.43-3.40 (m, 1H), 3.39 (m, 1H), 3.38-
3.34 (m, 3H), 3.28-3.22 (m, 4H), 3.17 (m, 2H), 2.56 (m, 6H),
2.40-2.32 (m, 3H), 2.06 (s, 2H), 1.73 (q, J ) 13.2, 1H), 0.81 (s,
3H), 0.79 (s, 3H). 13C NMR (D2O, 100 MHz): δ 174.4, 173.2,
172.2, 171.6, 170.6, 96.9, 79.3, 75.3, 75.2, 72.7, 71.8, 70.7, 70.6,
70.4, 68.9, 54.0, 48,8, 41.8, 39.8, 39.4, 38.5, 37.3, 35.0, 31.5, 30.5,
(18 mg, 41%). H NMR (D2O, 500 MHz, presat): δ 5.57 (d, J )
3.5, 1H), 3.94 (d, J ) 10.5, 1H), 3.84 (s, 1H), 3.80 (d, J ) 10.5,
1H), 3.79-3.70 (m, 3H), 3.54-3.30 (m, 9H), 3.30-3.22 (m, 4H,
3.21-3.13 (m, 3H), 2.56 (t, J ) 6.4, 2H), 2.40-2.30 (m, 3H), 1.72
(q, J ) 13.2, 1H), 0.82 (s, 3H), 0.79 (s, 3H). 13C NMR (D2O, 125
MHz): δ 174.4, 173.2, 172.2, 171.6, 170.6, 97.2, 79.3, 75.5, 75.2,
72.6, 72.0, 71.2, 70.6, 69.0, 54.0, 50.0, 48.8, 40.1, 39.4, 38.5, 35.6,
35.5, 35.0, 34.8, 31.5, 28.5, 20.7, 20.0. HRMS for C28H50N6O14S
(M + H) calcd 727.3246, found 727.3237. Purity, 98% (method
A, tR ) 3.47 min; method B, tR ) 3.25 min).
Compound 5e. D-Pantethine (0.27 g, 0.5 mmol) and lithium
acetoacetate (0.18 g, 1.5 mmol) were dissolved in DMF (10 mL).
The mixture was stirred for 10 min followed by the addition of
HATU (0.38 g, 1.0 mmol) and DIPEA (0.4 mL, 2.0 mmol). The
mixture was stirred for 16 h at room temperature before evaporation
under vacuum. The residue was triturated with dichloromethane
(2 × 10 mL) to yield a paste. This paste was dissolved in CHCl3/
MeOH (30 mL, 10/1, v/v) and washed with H2O (2 × 10 mL).
The organic layer was dried over sodium sulfate and evaporated to
dryness to give 20 as a yellowish powder. Compound 20 (80 mg,
∼0.1 mmol), DTT (15 mg, 0.1 mmol), and DIPEA (1 mL, 5 mmol)
were mixed in acetone/H2O (5 mL, 1/1 v/v). The mixture was
sonicated for 2 min and stirred overnight. This solution was
transferred to the N-6′-bromoacetylneamine solution (4 mL). The
resulting mixture was sonicated for 2 min and stirred for 1 h.
Workup was carried out as described for 5a and followed by HPLC
purification to afford 5e (4.6 mg, 10%). 1H NMR (D2O, 400 MHz,
presat): δ 5.57 (s, 1H), 3.87 (s, 1H), 3.79 (t, J ) 9.6, 2H), 3.69 (t,
J ) 9.6, 1H), 3.53-3.19 (m, 18 H), 2.62 (t, J ) 6.0, 2 H), 2.41-
2.34 (m, 3H), 2.16 (s, 3H), 1.71 (q, J ) 13.2, 1H), 0.80 (s, 3H),
0.77 (s, 3H). 13C NMR (D2O, 100 MHz): δ 215.2, 174.4, 174.0,
173.8, 173.5, 98.9, 88.5, 84.8, 76.2, 75.8, 73.2, 73.0, 72.0, 71.8,
69.3, 69.0, 55.8, 53.7, 51.2, 50.0, 48.8, 40.0, 39.6, 36.0, 35.8, 35.0,
32.0, 21.1, 19.3. HRMS for C29H53N6O13S (M + H) calcd 741.3386,
found 741.3384. Purity, 98% (method A, tR ) 3.35 min; method
B, tR ) 3.21 min).
Compound 6a. N-Acetyl-D-glucosamine (3 g, 13.5 mmol) was
dissolved in methanol (70 mL), and Amberlite IR 120 (H) resin (5
g) was added. The mixture was refluxed overnight, cooled to room
temperature, and filtered to remove the resin. The filtrate was
evaporated to dryness to give 6a as a white powder (2.83 g, 89%,
1
highly hygroscopic). H NMR (400 MHz, DMSO-d6): δ 7.71 (d,
J ) 4.0, 1H), 4.6 (br s, 3H), 4.50 (d, J ) 3.0, 1H), 3.62 (m, 2H),
3.38-3.46 (m, 2H), 3.28 (m, 1H), 3.22 (s, 3H), 3.10 (m, 1H), 1.82
(s, 3H). 13C NMR (100 MHz, DMSO-d6): δ 169.9, 98.6, 73.5, 71.6,
71.5, 61.3, 55.0, 54.5, 23.5. ESI-MS (M + H) for C9H17NO6, calcd
236.1, found 236.3.
Compound 6b. 6a (1 g, 4.25 mmol) was dissolved in pyridine
(15 mL), the mixture was cooled to -40 °C, and methanesulfonyl
chloride (0.35 mL, 4.5 mmol) was added. The mixture was stirred
at -40 °C for 2 h and at room temperature for 10 h. After
evaporation of the solvent, the residue was purified by flash
chromatography (Rf ) 0.40, CHCl3/MeOH 99/1) to give 6b as a
1
white powder (0.97 g, 73%). H NMR (DMSO-d6, 400 MHz): δ
7.77 (d, J ) 5.6, 1H), 5.36 (d, J ) 5.6, 1H), 4.88 (d, J ) 6.0, 1H),
4.54 (d, J ) 3.6, 1H), 4.38 (d, J ) 9.8, 1H), 4.27 (dd, J ) 10.8,
5.6, 1H), 3.66 (td, J ) 10.8, 3.6, 1H), 3.57 (dd, J ) 10.8, 5.6, 1H),
3.34 (s, 3H), 3.43 (m, 1H), 3.23 (s, 3H), 3.15 (m, 1H), 1.81 (s,
3H). 13C NMR (DMSO-d6, 100 MHz): δ 170.0, 98.8, 79.9, 71.2,
71.0, 70.4, 55.4, 54.3, 37.5, 23.4. ESI-MS (M + H) for C10H19-
NO8S calcd 314.3, found 314.2.
Compound 6c. 6b (0.63 g, 2.11 mmol) and NaN3 (0.68 g, 10.55
mmol) were mixed in acetone/H2O (30 mL, 3/2 v/v). The mixture
was refluxed for 16 h. After evaporation of the solvent, the residue