1974
C.-Y. Chang et al. / Bioorg. Med. Chem. Lett. 11 (2001) 1971–1974
phosphodiesterase enzyme source) was incubated with 1
3
The residue was eluted through a silica gel column with n-
hexane/EtOAc (9:1) to give pure compound 8 (19.5 g, 92%) as
mM cAMP (0.05 mCi [ H]cAMP) (Amersham) and var-
ious concentrations of 7 and Ro201724, a type 4 phos-
ꢁ
white prism crystals. Mp 57–58 C; MS (EI, 70eV): m/z 212
+
13
ꢁ
14 12 2
(M ); found C, 79.20; H, 5.59%. C H O requires C, 79.25;
phodiesterase inhibitor, for 30min at 37 C. Then the
formed AMP was converted to nucleotide by incubation
with Crotalus atrox snake venom (1 mg/mL in 0.2 M Tris–
H, 5.66%; UV, lmax (log e): 215.0(4.57), 251.0(4.46), 311.0
1
(
3.92); IR (KBr): 1685 (C¼O); H NMR (200 MHz, CDCl ): d
3
5.13 (2H, s, –OCH –), 7.23–7.32 (1H, m, H-4), 7.34–7.49 (8H,
m, H-2 , 3 , 4 , 5 , 6 , 2, 5, 6), 9.98 (1H, s, –CHO); C NMR
2
1
0
HCl, pH 8.0). Nucleoside product was separated
through AG 1-X8 resin (formate) (Bio-Rad) and radio-
activity was detected in a liquid scintillation counter. As
shown in Fig. 2, only Ro201724 suppressed the phospho-
diesterase activity. Compound 7 had no effect on cAMP
degradation. These results indicate that compound 7 ele-
vates cAMP levels through activation of adenylyl
cyclase but does not inhibit phosphodiesterase.
0
0
0
0
0
13
(
4
50MHz, CDCl ): d 0.01 (–OCH –), 113.07 (C-2), 121.97 (C-
3 2
0 0 0 0
), 123.45 (C-6), 127.32 (C-2 , 6 ), 127.99 (C-4 ), 128.46 (C-3 ,
0
0
5 ), 129.90(C-5), 136. 90 (C-1 ), 137.61 (C-1), 159.10(C-3),
191.85 (–CHO). 4-Benzyloxybenzaldehyde (9). 4-Hydroxy-
benzaldehyde (12.2 g, 100 mmol) was treated in the same
manner as described for compound 7. The residue was eluted
through a silica gel column with n-hexane/EtOAc (9:1) to give
pure compound 9 (19.5 g, 92%) as white needles. Mp 67–
ꢁ
+
6
8 C; MS (EI, 70eV): m/z 212 (M ); found C, 79.13; H,
There are many indirect adenylyl cyclase activators
which cause accumulation of cellular cAMP through the
influence on adenylyl cyclase-coupled receptors or sig-
naling pathways. Forskolin, the only known direct ade-
nylyl cyclase activator, stimulates adenylyl cyclase by
5
.56%. C14 requires C, 79.25, H, 5.66%; UV, lmax (log
12 2
H O
e): 214.0(4.6 )0 , 252.0(4.23), 315.0(3.91); IR (KBr): 1685
1
(
C¼O); H NMR (200 MHz, CDCl ): d 5.16 (2H, s, –OCH –),
3
2
0 0 0
7
.09 (2H, d, J=8.8 Hz, H-3, 5), 7.35–7.47 (5H, m, H-2 , 3 , 4 ,
5 , 6 ), 7.85 (2H, d, J=8.8 Hz, H-2, 6), 9.89 (1H, s, –CHO);
0
13
0
1
1
interaction with the catalytic subunit of the enzyme. As
an adenylyl cyclase activator, compound 7 has a chemi-
cal structure unlike forskolin and the known indirect
activators. Therefore, we recommend the use of com-
pound 7 as a new lead compound in the future develop-
ment of adenylyl cyclase activators. Currently, its
mechanism of action is under investigation and will be
reported separately.
C NMR (50MHz, CDCl
0
3
): d 70.05 (–OCH
2
–), 114.93 (C-3,
0 0
0
0
5), 127.26 (C-2 , 6 ), 128.11 (C-4 ), 128.51 (C-3 , 5 ), 129.91 (C-
0
1
2
1
), 131.78 (C-2, 6), 135.73 (C-1 ), 163.52 (C-4), 190.57 (–CHO).
-Benzyloxyacetophenone (10). 2-Hydroxyacetophenone (13.6 g,
00mmol) was treated in the same manner as described for
compound 7. The residue was eluted through a silica gel col-
umn with CHCl /n-hexane (1:2) to give pure compound 10
3
ꢁ
(
(
15.8 g, 70%) as colorless prism crystals. Mp 41–42 C; MS
+
EI, 70eV): m/z 226 (M ); found C, 79.58; H, 6.11%.
(log e): 211.0
C H O requires C, 79.65, H, 6.19%; UV, l
1
5
14
2
max
1
(4.57), 246.0 (4.10), 305.0 (3.80); IR (KBr): 1665 (C¼O); H
NMR (200 MHz, CDCl
OCH –), 7.02–7.06 (2H, m, H-3, 5), 7.39–7.50 (6H, m, H-2 ,
2
References and Notes
3
): d 2.63 (3H, s, –CH
3
), 5.17 (2H, s,
0
–
0
0
0
0
13
1
. Babior, B. M. Am. J. Med. 2000, 109, 33.
3 , 4 , 5 , 6 , 4), 7.78 (1H, dd, J=8.0, 2.0 Hz, H-6); C NMR
), 70.40 (–OCH
2. Sunahara, R. K.; Dessauer, C. W.; Gilman, A. G. Annu.
Rev. Pharmacol. Toxicol. 1996, 36, 461.
(50MHz, CDCl
(C-3), 120.66 (C-5), 127.38 (C-2 , 6 ), 128.05 (C-4 ), 128.42 (C-
3
): d 31.98 (–CH
3
0
2
–), 112.58
0
0
0
0
0
3
2
4
. Houslay, M. D.; Milligan, G. Trends Biochem. Sci. 1997,
2, 217.
. Cronstein, B. N.; Levin, R. I.; Philips, M.; Hirschhorn, R.;
1), 128.51 (C-3 , 5 ), 130.27 (C-6), 133.47 (C-4), 135.98 (C-1 ),
157.82 (C-2), 199.76 (–COCH ). 2-Benzyloxybenzamide (11).
2-Hydroxybenzamide (13.7 g, 100 mmol) was treated in the
3
Abramson, S. B.; Weissmann, G. J. Immunol. 1992, 148, 2201.
. Cronstein, B. N.; Rosenstein, E. D.; Kramer, S. B.; Weiss-
mann, G.; Hirschhorn, R. J. Immunol. 1985, 135, 1366.
same manner as described for compound 7. The residue was
eluted through a silica gel column with CHCl to give pure
5
3
ꢁ
compound 11 (21.6 g, 95%) as white needles. Mp 114–115 C;
+
6
. Richter, J. J. Leukoc. Biol. 1992, 51, 270.
. Experimental details for synthesis: 2-Benzyloxybenzaldehyde
MS (EI, 70eV): m/z 227 (M ); found C, 73.85; H, 5.65; N,
6.14%. C H NO requires C, 74.00; H, 5.73; N, 6.17%; UV,
7
1
4
13
2
(
7). 2-Hydroxybenzaldehyde (12.2 g, 100 mmol) was dissolved
in acetone (150mL). Benzyl chloride (15 mL, 130mmol),
anhydrous K CO (20g) and anhydrous KI (26 g) were added.
lmax (log e): 210.0 (4.57), 225.0 (4.36), 290.0 (3.87); IR (KBr):
1
1647 (C¼O), 3406 (NH
(2H, s, –OCH –), 6.08 (1H, br, N-Ha), 7.05–7.14 (2H, m, H-3,
5), 7.38–7.52 (6H, m, H-2 , 3 , 4 , 5 , 6 , 4), 7.75 (1H, br, N-
2 3
); H NMR (200 MHz, CDCl ): d 5.19
2
3
2
0
0
0
0
0
The mixture was stirred under reflux (50–60) for 6 h, and then
evaporated in vacuo. Water (100 mL) was added and the mix-
ture was extracted with CHCl . The separated organic layer
4
was dried with MgSO , and then evaporated in vacuo. Finally
the residue was eluted through a silica gel column with n-hex-
ane/EtOAc (4:1) to give pure compound 7 (18.4 g, 87%) as
1
3
Hb), 8.25 (1H, dd, J=7.8, 1.9 Hz, H-6); C NMR (50MHz,
CDCl ): d 71.06 (–OCH –), 112.45 (C-3), 120.92 (C-1), 121.29
(C-5), 127.64 (C-2 , 6 ), 128.49 (C-4 ), 128.75 (C-3 , 5 ), 132.46
3
3
2
0
0
0
0
0
0
(C-6), 133.12 (C-4), 135.34 (C-1 ), 156.90(C-2), 166.78
2
(–CONH ).
ꢁ
pale orange prism crystals. Mp 48–49 C; MS (EI, 70eV): m/z
8. Wang, J. P.; Raung, S. L.; Kuo, Y. H.; Teng, C. M. Eur. J.
Pharmacol. 1995, 288, 341.
9. Markert, M.; Andrews, P. C.; Babior, B. M. Meth. Enzy-
mol. 1984, 105, 358.
10. Wright, C. D.; Kuipers, P. J.; Kobylarz-Singer, D.;
Devall, L. J.; Klinkefus, B. A.; Weishaar, R. E. Biochem.
Pharmacol. 1990, 40, 699.
11. Seamon, K. B.; Daly, J. W. Adv. Cyclic Nucleotide Protein
Phosph. Res. 1986, 20, 1.
+
2
7
3
12 (M ); found C, 79.17; H, 5.54%. C H O requires C,
14 12 2
9.25; H, 5.66%; UV, lmax (log e): 215.4 (4.46), 253.6 (4.00),
1
18.2 (3.89); IR (KBr): 1685 (C¼O); H NMR (200 MHz,
CDCl
7
3
): d 5.20(2H, s, –OCH
.35–7.48 (5H, m, H-2 , 3 , 4 , 5 , 6 ), 7.54 (1H, ddd, J=8.1,
.7, 1.8 Hz, H-4), 7.87 (1H, dd, J=8.0, 2.0 Hz, H-6), 10.58
2
–), 7.01—7.09 (2H, m, H-3, 5),
0
0
0
0
0
7
(
1H, s, –CHO); 13C NMR (50MHz, CDCl
): d 70.24 (–OCH
–),
0
3
2
0
1
1
1
12.82 (C-3), 120.80 (C-5), 124.96 (C-1), 127.08 (C-2 , 6 ),
0
0
0
28.06 (C-4 ), 128.23 (C-6), 128.52 (C-3 , 5 ), 135.70(C-4),
0
12. Beavo, J. A.; Conti, M.; Heaslip, R. J. Mol. Pharmacol.
1994, 46, 39.
35.86 (C-1 ), 160.84 (C-2), 189.54 (–CHO). 3-Benzyloxy-
benzaldehyde (8). 3-Hydroxybenzaldehyde (12.2 g, 100 mmol)
was treated in the same manner as described for compound 7.
13. Reeves, M. L.; Leigh, B. K.; England, P. J. Biochem. J.
1987, 241, 535.