L. Lu et al. / Bioorg. Med. Chem. 17 (2009) 7548–7561
7559
HRMS (FAB) (m/z) calcd for C23H37N4O4S+ (MH+) 465.2530, found
465.2524.
system was stirred 20 h under Ar. After evaporation of solvent, the
crude product was purified by silica gel chromatography (10%
methanol in chloroform, TLC: Rf = 0.2) to give 40 mg (69%) of pure
23. 1H NMR (CDCl3, 400 MHz), d 7.70 (d, J = 7.6 Hz, 2H), 7.52 (d,
J = 7.6 Hz, 2H), 7.35 (dd, J = 7.6, 7.6 Hz, 2H), 7.25 (dd, J = 7.6,
7.6 Hz, 2H), 6.58 (m, 1H), 5.94 (m, 1H), 5.74 (m, 1H), 5.28 (m,
2H), 4.31 (d, J = 6.8 Hz, 2H), 4.14 (t, J = 6.8 Hz, 1H), 4.03–3.60
(9H), 2.82–2.78 (t, J = 7.2 Hz, 2H), 2.69–2.65 (t, J = 7.2 Hz, 2H),
2.21–2.17 (t, J = 7.2 Hz, 4H), 1.54–1.45 (m, 2H), 1.23–1.19 (24H),
0.86 (t, J = 6.8 Hz, 3H). 13C NMR (CDCl3, 100 MHz), d 173.69 (CO),
173.42 (CO), 143.69 (C), 141.22 (C), 131.07 (CH), 127.77 (CH),
127.09 (CH), 125.02 (CH), 120.97 (CH), 119.98 (CH), 107.20 (CH),
105.34 (CH), 77.20 (CH) 69.07 (CH2), 66.66 (CH2), 65.37 (CH2),
64.30 (CH2), 46.65 (CH2), 46.35 (CH), 33.97 (CH2), 33.11 (CH2),
31.92 (CH2), 29.73 (CH2), 29.67 (CH2), 29.58 (CH2), 29.37 (CH2),
29.23 (CH2), 24.83 (CH2), 22.69 (CH2), 21.23 (CH2), 14.11 (CH3).
HRMS (FAB) (m/z) calcd for C41H58NNaO9P+ (MNa+) 776.3904,
found 776.3939; calcd for C41H57NNa2O9P+ (MNa2+) 798.3724,
found 798.3717.
5.31. Octadec-9-enoic acid 2-[(2-{2-[2-(9H-fluoren-9-
ylmethoxycarbonyl)-ethyl]-pyrrol-1-yl}-ethoxy)-hydroxy-
phosphoryloxy]-1-hexadecanoyloxymethyl-ethyl ester (21)
Triethylamine (TEA) (25
(50 mg, 0.065 mmol) in 500
0.065 mmol) in 500 L CHCl3 was added to the mixture. The sys-
lL, 0.247 mmol) was added to POPE
lL CHCl3, then DOHAFm (22 mg,
l
tem was stirred 20 h under Argon. After evaporation of solvent,
the crude product was purified by silica gel chromatography
(10% methanol in chloroform, TLC: Rf = 0.2) to give 50 mg (76%)
of pure 21. 1H NMR (CD3OD/CDCl3 = 1:1, 400 MHz), d 7.74 (d,
J = 7.6 Hz, 2H), 7.53 (d, J = 7.6 Hz, 2H), 7.36 (dd, J = 7.6, 7.6 Hz,
2H), 7.27 (dd, J = 7.6, 7.6 Hz, 2H), 6.63 (dd, J = 2.8, 1.6 Hz, 1H),
5.94 (dd, J = 3.2, 2.8 Hz, 1H), 5.77 (m, 1H), 5.28 (m, 2H), 4.36 (d,
J = 6.8 Hz, 2H), 4.28 (m, 1H), 4.17 (t, J = 6.8 Hz, 1H), 4.05–3.98
(6H), 3.68 (t, J = 6.4 Hz, 2H), 2.86–2.82 (t, J = 7.2 Hz, 2H), 2.73–
2.69 (t, J = 7.2 Hz, 2H), 2.25–2.21 (4H), 2.0–1.94 (4H), 1.57 (4H),
1.21 (44H), 0.84 (t, J = 6.4 Hz, 6H). 13C NMR (CD3OD/CDCl3 = 1:1,
50 MHz, APT), d 175.21 (+) (CO), 174.84 (+) (CO), 174.81 (+) (CO),
145.08 (+) (C), 142.68 (+) (C), 132.30 (+) (C), 131.24 (ꢂ) (CH),
130.95 (ꢂ) (CH), 129.13 (ꢂ) (CH), 128.53 (ꢂ) (CH), 126.31 (ꢂ)
(CH), 121.30 (ꢂ) (CH), 121.22 (ꢂ) (CH), 108.49 (ꢂ) (CH), 106.77
(ꢂ) (CH), 71.86 (ꢂ) (CH), 67.96 (+) (CH2), 66.62 (+) (CH2), 64.60
(+) (CH2), 64.05 (+) (CH2), 48.10 (+) (CH2), 47.82 (ꢂ) (CH), 35.47
(+) (CH2), 35.34 (+) (CH2), 34.66 (+) (CH2), 33.27 (+) (CH2), 31.03
(+) (CH2), 30.85 (+) (CH2), 30.70 (+) (CH2), 30.66 (+) (CH2), 30.61
(+) (CH2), 30.49 (+) (CH2), 28.49 (+) (CH2), 26.24 (+) (CH2), 26.20
(+) (CH2), 23.98 (+) (CH2), 22.64 (+) (CH2), 15.14 (ꢂ) (CH3). HRMS
(FAB) (m/z) calcd for C60H91NNa2O10P+ [(MꢂH)Na2+] 1062.6177,
found 1062.6.
5.34. Hexadecanoic acid 3-([2-[2-(carboxyethyl)-pyrrol-1-yl]-
ethoxy]-hydroxy-phosphoryloxy)-2-hydroxy-propyl ester
(lysoPE-CEP, 24)
DBU (20
lL, 0.13 mmol) was added to 23 (24 mg, 0.03 mmol) in
500 L CHCl3. After 3 h, the reaction was completed and the sys-
l
tem was diluted by 1.5 mL CHCl3. The solution was washed with
2 mL phosphate buffer of pH 5.5. The organic part was washed
with brine, dried over magnesium sulfate and vacuum-filtered,
concentrated, purified by flash chromatography (CHCl3/MeOH/
H2O = 65:25:4, v/v, TLC: Rf = 0.18) to yield 16 mg (95%) of pure
lysoPE-CEP. 1H NMR (CD3OD/CDCl3 = 1:1, 400 MHz), d 6.60 (m,
1H), 5.96 (m, 1H), 5.83 (m, 1H), 4.10–4.0 (5H), 3.74–3.6 (m, 2H),
3.6–3.5 (m, 2H), 2.8–2.9 (m, 2H), 2.68–2.65 (m, 2H), 2.32 (t,
J = 6.8 Hz, 2H), 1.57 (m, 2H), 1.23 (24H), 0.85 (t, J = 7.2 Hz, 3H).
13C NMR (CD3OD/CDCl3/D2O = 50:50:1, 100 MHz), d 175.20 (CO),
133.00 (C), 121.23 (CH), 107.36 (CH), 105.56 (CH), 70.80 (CH),
66.05 (CH2), 65.63 (CH2), 47.17 (CH2), 34.61 (CH2), 33.29 (CH2),
32.48 (CH2), 30.22 (CH2), 30.05 (CH2), 29.90 (CH2), 29.88 (CH2),
29.72 (CH2), 27.04 (CH2), 25.42 (CH2), 23.21 (CH2), 14.38 (CH3).
HRMS (FAB) (m/z) calcd for C27H48NNaO9P+ (MNa+) 598.3121,
found 598.3053.
5.32. Octadec-9-enoic acid 2-({2-[2-(2-carboxy-ethyl]-pyrrol-1-
yl}-ethoxy)-hydroxy-phosphoryloxy]-1-hexadecanoyloxym-
ethyl-ethyl ester (PE-CEP, 22)
DBU (20
lL, 0.13 mmol) was added to 21 (24 mg, 0.025 mmol)
in 500 L CHCl3. After 3 h, the reaction was completed and the sys-
l
tem was diluted by 1.5 mL CHCl3. The solution was washed with
2 mL phosphate buffer of pH 5.5. The organic phase was washed
with brine, dried over magnesium sulfate, then vacuum-filtered,
concentrated, purified by flash chromatography (15% methanol in
chloroform, TLC: Rf = 0.18) to yield 19 mg (90%) of pure 22. 1H
NMR (CD3OD/CDCl3 = 1:1, 400 MHz), d 6.56 (m, 1H), 5.96 (m, 1H),
5.83 (m, 1H), 5.30 (m, 2H), 5.15 (m, 1H), 4.33 (m, 1H), 4.05–3.98
(6H), 3.81 (m, 2H), 2.83 (m, 2H), 2.58 (m, 2H), 2.27 (t, J = 7.2 Hz,
4H), 2.0–1.94 (4H), 1.57 (4H), 1.21 (44H), 0.84 (t, J = 6.4 Hz, 6H).
13C NMR (CD3OD/CDCl3 = 1:1, 100 MHz), d 180.36 (COOH), 174.49
(CO), 174.08 (CO), 133.54 (C), 130.46 (CH), 130.14 (CH), 120.98
(CH), 107.26 (CH), 105.05 (CH), 71.00 (CH), 65.84 (CH2), 64.03
(CH2), 63.11 (CH2), 47.29 (CH2), 34.70 (CH2), 34.56 (CH2), 32.40
(CH2), 32.42 (CH2), 30.23 (CH2), 30.17 (CH2), 30.00 (CH2), 29.80
(CH2), 29.63 (CH2), 27.66 (CH2), 25.40 (CH2), 23.13 (CH2), 14.31
(CH3). HRMS (FAB) (m/z) calcd for C46H82NNaO10P+ (MNa+)
862.5574, found 862.5550; calcd for C46H81NNa2O10P+ [(MꢂH)-
Na2+] 884.5394, found 884.5291.
5.35. 2-(9H-Fluoren-9-ylmethoxycarbonylamino)-6-{2-[2-(9H-
fluoren-9-ylmethoxycarbonyl)-ethyl]-pyrrol-1-yl}-hexanoic
acid (25)
6-Amino-2-(9H-fluoren-9-ylmethoxycarbonyl amino)-hexanoic
acid (40 mg, 0.1 mmol) was suspended in 10 mL methanol with
DOHAFm (30 mg, 0.089 mmol). Then 20 lL of acetic acid was
added. The suspension dissolved gradually and a light yellow oil
was generated at the bottom of the flask as the reaction proceeded.
The system was stirred 24 h under Ar. After the removal of solvent,
the crude compound was purified by silica gel chromatography (4%
methanol in chloroform) to give 45 mg (75%) of light yellow oil 25.
TLC (4% methanol in chloroform): Rf = 0.15; 1H NMR (CDCl3,
400 MHz), d 7.75 (d, J = 7.2 Hz, 4H), 7.55 (d, J = 7.6 Hz, 4H), 7.39
(dd, J = 7.2, 7.6 Hz, 4H), 7.30 (dd, J = 7.2, 7.6 Hz, 4H), 6.57 (m, 1H),
6.06 (dd, J = 3.2, 3.2 Hz, 1H), 5.87 (m, 1H), 5.4 (m, 1H), 4.41 (d,
J = 6.8 Hz, 2H), 4.39 (m, 1H), 4.20 (t, J = 6.8 Hz, 2H), 3.78 (t,
J = 6.8 Hz, 2H), 2.85–2.73 (4H), 1.91–1.39 (6H). 13C NMR (CDCl3,
100 MHz), d 176.58 (COOH), 173.08 (COO), 156.04 (CO), 143.75
(C), 143.66 (C), 141.25 (C), 130.58 (C), 127.75 (CH), 127.69 (CH),
127.07 (CH), 127.03 (CH), 125.01 (CH), 124.95 (CH), 119.99 (CH),
106.99 (CH), 105.23 (CH), 67.04 (CH2), 66.46 (CH2), 53.49 (CH),
5.33. Hexadecanoic acid 3-[(2-{2-[2-(9H-fluoren-9-ylmeth-
oxycarbonyl)-ethyl]-pyrrol-1-yl}-ethoxy)-hydroxy-phos-
phoryloxy]-2-hydroxy-propyl ester (23)
Triethylamine (TEA) (12
(42 mg, 0.093 mmol) in 500
0.077 mmol) in 500 L CHCl3 was added to the mixture. The cloudy
l
L, 0.116 mmol) was added to lyso-PE
lL CHCl3, then DOHAFm (26 mg,
l