9764
T. Kudoh et al. / Tetrahedron 64 (2008) 9754–9765
reaction mixture was diluted with EtOAc and washed with H2O. The
water layer was evaporated and co-evaporated with toluene
(3 mLꢃ3) to recover 11. The organic layer was washed with brine,
dried (Na2SO4), and evaporated. A mixture of the residue and K2CO3
(240 mg, 1.74 mmol) in DMF (9 mL) was stirred at 50 ꢀC for 30 min.
The reaction mixture was diluted with EtOAc and washed with H2O
and brine, dried (Na2SO4), and evaporated. The residue was purified
by column chromatography (SiO2, 75% EtOAc in hexane) to give 10b
125.6, 125.3, 122.2, 114.7, 112.4, 90.8, 84.8, 84.7, 84.5, 84.4, 84.1, 81.0,
66.8, 66.2, 60.3, 59.9, 27.3, 27.1, 25.7, 25.3; 31P NMR (CDCl3,
202 MHz) d 50.8 (s); HRMS (FAB, positive) calcd for C34H38N4O19S2:
741.1818 (MHþ), found: 741.1831.
4.1.14. N-1-{(1R,2S,3R)-2,3-Isopropylidenedioxy-4-
(phosphonoxymethyl)cyclopent-4-en-1-yl}-50-O-
{(phenylthio)phospholyl}-20,30-O-isopropylideneinosine (9b)
Compound 9b (oil, 26 mg, 36%) was obtained from 19b (84 mg,
0.11 mmol) as described for the synthesis of 9a. 1H NMR (D2O,
(179 mg, 35%) as a brown foam. 1H NMR (CDCl3, 500 MHz)
d 8.01 (s,
1H), 7.87 (s, 1H), 6.09 (d, 1H, J¼2.6 Hz), 5.64 (m, 2H), 5.33 (d, 1H,
J¼5.7 Hz), 5.03 (dd, 1H, J¼2.6, 6.0 Hz), 4.88 (dd, 1H, J¼6.0, 2.3 Hz),
4.69 (d, 1H, J¼5.7 Hz), 4.51 (m, 2H), 4.41 (m, 3H), 1.61, 1.45, 1.38, 1.33
(each s, each 3H), 0.85 (s, 9H), 0.062, 0.036 (each s, each 3H); 13C
500 MHz) d 8.22 (s, 1H), 8.14 (s, 1H), 7.08–7.22 (m, 5H), 6.30 (m, 1H),
5.91 (m, 1H), 5.59 (m, 1H), 5.49 (d, 1H, J¼5.9 Hz), 5.36 (d, 1H,
J¼5.7 Hz), 5.10 (d, 1H, J¼5.9 Hz), 4.67 (m, 1H), 4.64 (m, 2H), 4.38 (d,
1H, J¼5.7 Hz), 4.29 (m,1H), 4.10 (m,1H), 3.18 (q,12H, J¼7.3 Hz),1.64,
1.51, 1.42, 1.39 (each s, each 3H), 1.26 (t, 18H, J¼7.3 Hz); 13C NMR
NMR (CDCl3, 125 MHz)
d 156.3, 151.9, 146.8, 145.4, 138.5, 123.7,
122.9, 114.2, 112.5, 91.4, 87.0, 85.5, 84.4, 84.3, 81.3, 63.5, 62.8, 59.9,
27.3, 27.2, 25.9, 25.3,18.3, ꢁ5.5, ꢁ5.6; HRMS (FAB, positive) calcd for
(D2O, 125 MHz) d 157.9, 149.2, 149.1, 147.7, 146.6, 141.4, 132.8, 132.8,
C
28H43N4O8Si: 591.2850 (MHþ), found: 591.2862; UV (MeOH) lmax
129.9, 129.4, 128.2, 124.2, 115.2, 113.6, 91.2, 86.5, 86.4, 84.9, 84.2,
245 nm.
83.8, 81.8, 67.0, 66.3, 62.2, 47.1, 26.6, 26.4, 25.3, 24.7, 8.7; 31P NMR
(D2O, 202 MHz)
C
lmax 244 nm.
d 17.4 (s), 1.0 (s); HRMS (FAB, negative) calcd for
4.1.11. N-1-{(1R,2S,3R)-4-Dimethoxytrityloxymethyl-2,3-
isopropylidenedioxycyclopent-4-en-1-yl}-20,30-O-
isopropylideneinosine (17b)
28H33N4O13P2S: 727.1246 [(MꢁH)ꢁ], found: 727.1230; UV (H2O)
Compound 17b (oil, 158 mg, 61%) was obtained from 10b
4.1.15. 400,600-Didehydro-cyclic IDP-carbocyclic-ribose
diacetonide (20b)
(195 mg, 0.330 mmol) as described for the synthesis of 17a. 1H NMR
(CDCl3, 500 MHz)
d
8.00 (s, 1H), 7.86 (s, 1H), 6.83–7.48 (m, 13H),
Compound 20b (13 mg, 57%) was obtained from 9b (amorphous
5.93 (d, 1H, J¼4.1 Hz), 5.90 (m, 1H), 5.83 (m, 1H), 5.17 (d, 1H,
J¼5.6 Hz), 5.09 (m, 1H), 5.07 (m, 1H), 4.61 (d, 1H, J¼5.6 Hz), 4.50 (m,
1H), 4.00 (m, 1H), 3.96 (m, 1H), 3.82 (m, 1H), 3.78 (s, 6H), 3.75 (m,
1H), 1.64, 1.39, 1.36, 1.27 (each s, each 3H); 13C NMR (CDCl3,
solid, 26 mg, 28
(D2O, 500 MHz)
m
d
mol) as described for the synthesis of 20a. 1H NMR
8.45 (s,1H), 8.19 (s,1H), 6.30 (m,1H), 6.22 (m,1H),
5.72 (d, 1H, J¼6.1 Hz), 5.61 (m, 1H), 5.55 (dd, 1H, J¼6.1, 3.0 Hz), 5.48
(d, 1H, J¼5.2 Hz), 4.93 (m, 1H), 4.60 (d, 1H, J¼5.2 Hz), 4.53 (m, 1H),
4.46 (m, 1H), 3.98 (m, 1H), 3.81 (m, 1H), 3.19 (q, 12H, J¼7.3 Hz), 1.62,
1.46, 1.45, 1.38 (each s, each 3H), 1.27 (t, 18H, J¼7.3 Hz); 13C NMR
125 MHz) d 171.0, 158.5, 156.0, 151.4, 146.2, 145.1, 144.6, 139.7, 135.9,
135.7, 132.8, 132.0, 132.0, 131.9, 131.9, 129.9, 129.8, 128.5, 128.4,
127.9, 127.8, 126.8, 125.7, 121.4, 114.2, 113.2, 112.5, 93.2, 86.6, 86.2,
84.6, 84.1, 81.5, 66.3, 62.9, 61.2, 60.3, 55.1, 27.4, 26.0, 25.2, 21.0, 14.1;
HRMS (FAB, positive) calcd for C43H47N4O10: 779.3292 (MHþ),
found: 779.3284; UV (MeOH) lmax 272, 265 nm.
(D2O, 125 MHz) d 158.8, 149.5, 148.1, 146.1, 142.6, 128.4, 124.6, 115.0,
113.3, 91.5, 86.7, 84.7, 83.8, 82.4, 81.5, 65.3, 64.3, 62.4, 47.1, 26.8,
26.4, 25.7, 24.7, 8.7; 31P NMR (D2O, 202 MHz)
d
ꢁ10.5 (d, J¼15.3 Hz),
ꢁ10.7 (d, J¼15.3 Hz); HRMS (FAB, negative) calcd for
C
22H27N4O13P2: 617.1055 [(MꢁH)ꢁ], found: 617.1049; UV (H2O) lmax
4.1.12. N-1-{(1R,2S,3R)-4-Dimethoxytrityloxymethyl-2,3-
isopropylidenedioxycyclopent-4-en-1-yl}-50-O-{bis(phenylthio)-
phosphoryl}-20,30-O-isopropylideneinosine (18b)
251 nm.
4.1.16. 400,600-Didehydro-cyclic IDP-carbocyclic-ribose (8b)
Compound 18b (oil, 149 mg, 71%) was obtained from 17b
Compound 8b (amorphous solid,10 mg, 82%) was obtained from
20b (13 mg, 16
(D2O, 500 MHz) d 8.35 (s, 1H), 8.19 (s, 1H), 6.22 (m, 1H), 5.99 (d, 1H,
(158 mg, 0.203 mmol) as described for the synthesis of 18a. 1H NMR
m
mol) as described for the synthesis of 8a. 1H NMR
(CDCl3, 500 MHz)
d
7.84 (s, 1H), 7.79 (s, 1H), 6.82–7.68 (m, 23H),
6.06 (d, 1H, J¼2.5 Hz), 5.88 (m, 1H), 5.82 (m, 1H), 5.15 (dd, 1H, J¼2.5,
6.3 Hz), 5.10 (d, 1H, J¼5.6 Hz), 4.96 (dd, 1H, J¼6.3, 3.1 Hz), 4.52 (d,
1H, J¼5.6 Hz), 4.46 (m, 1H), 4.39 (m, 2H), 3.99 (d, 1H, J¼15.5 Hz),
3.78 (s, 6H), 3.77 (d, 1H, J¼15.5 Hz), 1.62, 1.40, 1.37, 1.30 (each s, each
J¼6.2 Hz), 5.41 (m, 1H), 5.32 (dd, 1H, J¼6.2, 4.8 Hz), 4.88 (m, 1H),
4.81 (d, 1H, J¼4.0 Hz), 4.59–4.71 (m, 3H), 4.32 (m, 1H), 4.14 (d, 1H,
J¼4.0 Hz), 3.99 (m, 1H), 3.19 (m, 6H), 1.27 (m, 9H); 13C NMR (D2O,
125 MHz)
d 159.1, 151.5, 148.5, 144.9, 143.4, 126.3, 125.1, 91.1, 85.1,
3H); 13C NMR (CDCl3, 125 MHz)
d
171.1, 158.6, 158.5, 156.1, 150.9,
75.4, 73.3, 73.1, 71.2, 66.0, 64.9, 61.9, 47.1, 8.7; 31P NMR (D2O,
146.6, 145.4, 144.7, 139.0, 136.0, 135.8, 135.3, 135.2, 135.1, 135.0,
132.1,132.1,132.0,131.9,131.9,129.9,129.9,129.7,129.5,129.4,128.5,
128.4, 128.0, 127.9, 126.9, 125.7, 125.3, 121.9, 114.8, 113.2, 112.3, 90.8,
86.7, 84.8, 84.6, 84.4, 84.1, 80.9, 66.0, 61.2, 60.3, 55.2, 27.4, 27.1, 26.0,
202 MHz)
d
ꢁ9.9 (d, J¼15.3 Hz), ꢁ10.5 (d, J¼15.3 Hz); HRMS (FAB,
negative) calcd for C16H19N4O13P2: 537.0429 [(MꢁH)ꢁ], found:
537.0403; UV (H2O) lmax 251 nm.
25.3, 21.0, 14.2; 31P NMR (CDCl3, 202 MHz)
d
50.7 (s); HRMS (FAB,
4.2. Biological evaluations with sea urchin egg homogenate,
neuronal cells, and T cells
positive) calcd for C55H56N4O11PS2: 1043.3125 (MHþ), found:
1043.3132.
These bioassays were carried out as reported previously.7c
4.1.13. N-1-{(1R,2S,3R)-4-Hydroxymethyl-2,3-isopropylidene-
dioxycyclopent-4-en-1-yl}-50-O-{bis(phenylthio)-
4.2.1. NMR titrations
phospholyl}-20,30-O-isopropylideneinosine (19b)
NMR titrations were carried out as previously reported.29 The
experiments were performed in two steps in which 0.50 cm3 of the
same initial solution of studied compounds of 3.0ꢃ10ꢁ3 mol dmꢁ3
in 2H2O was successively subjected to potentiometric and NMR
titrations. It should be noted that the glass electrode was calibrated
in a concentration scale and the measurements done in 2H2O, so
that here pH means the cologarithm of the concentration of 2Hþ.
The processing of the pH measurements allowed the total con-
centration of the ligand and the acid as well as the macroscopic
Compound 19b (oil, 84 mg, 79%) was obtained from 18b
(149 mg, 0.143 mmol) as described for the synthesis of 19a. 1H NMR
(CDCl3, 500 MHz) d 7.86 (s, 1H), 7.83 (s, 1H), 7.27–7.67 (m, 10H), 6.05
(d, 1H, J¼2.5 Hz), 5.65 (m, 1H), 5.62 (m, 1H), 5.26 (d, 1H, J¼5.7 Hz),
5.13 (dd, 1H, J¼2.5, 6.3 Hz), 4.95 (dd, 1H, J¼6.3, 3.1 Hz), 4.59 (d, 1H,
J¼5.7 Hz), 4.45 (m, 1H), 4.40 (m, 4H), 1.61, 1.43, 1.36, 1.30 (each s,
each 3H); 13C NMR (CDCl3, 125 MHz)
d 156.2, 152.2, 146.7, 145.6,
139.2, 135.3, 135.3, 135.1, 135.1, 129.8, 129.5, 128.5, 128.4, 125.7,