170
7.10–7.30 (6H, m), 8.25 (1H, d), 8.50 (1H, s). 13C NMR (DMSO)
23.16, 25.09, 35.41, 37.10, 48.50, 58.20, 65.75, 114.25, 116.86,
120.0, 121.92, 125.87, 128.36, 128.58, 128.65, 136.81, 141.79,
144.14, 161.05, 166.79. HRMS 381.2146 (M + H). C23H29N2O3 re-
quires 381.2178.
(2H, s), 5.32–5.36 (1H, m), 6.55 (1H, d), 7.10–7.50 (10H, m),
7.60–7.65 (2H, m), 7.92 (1H, d). HRMS 429.2188 (M + H).
C27H29N2O3 requires 429.2178.
5-[2-(2-Methyl-5-phenylpent-2-yl)amino-1-hydroxyethyl]-8-benzyl-
oxy-2(1H)-quinolinone (10f). The yield was 0.9 g (64%). Recrystalli-
zation from 2-propanol gave analytically pure product. M.p.
224–225°C. 1H NMR (DMSO): 1.32 (6H, s), 1.68 (4H, m), 2.59 (2H,
m), 2.90 (2H, m), 5.40 (2H, s), 5.60 (1H, m), 6.25 (1H, s), 6.63 (1H,
d), 7.22–7.42 (9H, m), 7.63 (2H, d), 8.47 (1H, d), 9.80 (1H, broad sig-
nal), 10.8 (1H, s). 13C NMR (DMSO): 26.63, 24.93, 35.21, 36.63,
48.03, 59.02, 65.14, 69.67, 112.14, 116.48, 119.46, 122.52, 125.78,
127.78, 127.90, 128.25, 128.35, 129.40, 130.37, 136.35, 136.60,
141.61, 143.74, 160.94. HRMS 471.2641 (M + H). C30H35N2O3 re-
quires 471.2648.
5-[2-(2-Propylamino)-1-hydroxyethyl]-8-hydroxy-2(1H)-quinolinone
1
(11 g). The yield was 150 mg (71%). M.p. 145–149°C. H NMR
(MeOD): 1.34 (6H, s), 3.19 (2H, s), 3.40–3.44 (1H, m), 5.42 (2H, s),
6.55–6.57 (1H, m), 6.99–7.05 (1H, m), 7.23–7.25 (1H, m), 8.27–8.29
(1H, m), 8.53 (1H, s). 13C NMR (MeOD): 19.0, 52.0, 67.36, 67.44,
115.40, 118.96, 122.16, 129.29, 129.35, 129.40, 138.66, 145.48,
163.85. HRMS 263.1391 (M + H). C14H19N2O3 requires 263.1395.
Synthesis of catecholamine derivative (Fig. 2)
5-[2-(2-Propylamino)-1-hydroxyethyl]-8-benzyloxy-2(1H)-quinol-
inone (10g). The yield was 0.48 g (65.5%). M.p. 122–123°C. HRMS
353.1895 (M + H). C21H25N2O3 requires 353.1865.
The synthetic scheme for the catecholamine epoxide 12 was per-
formed according to the procedure described by Marki et al. (1988).
The epoxide was reacted neat with phenteramine and the adduct puri-
fied by column chromatography. In the final step, the two benzyl pro-
tecting groups were removed by hydrogenolysis using 1,4-cyclo-
hexadiene as hydrogen source.
General procedure for preparation of ligands (11a–11fg; Fig. 1)
The protected aminoalcohols (10a–10f) were dissolved in methanol
and ammonium formate (excess) followed by palladium on charcoal
(10%) added at once under nitrogen. The reaction was heated until no
more starting material was detected. The mixture was then cooled to
room temperature, filtered through celite and the filtrate concentrated
under vacuum. Products were purified using preparative thin layer
chromatography on silica gel using a mixture of methylene chloride-
methanol-ammonium hydroxide as eluent unless specified otherwise.
1-{[2-[(1-Phenyl-2-methylprop-2-yl)amino]-1-hydroxyethyl]-3,4-bis
(benzyloxy)}-benzene (13). 1,2-Bis(benzyloxy)-4-oxiranylbenzene
12 (2.89 g, 9.4 mmol) was reacted with freshly prepared phen-
teramine (3 ml, excess) and the mixture was heated under nitrogen for
6 h. The reaction was monitored by TLC, and when epoxide was no
longer detected, the mixture was diluted with methylene chloride (50
ml) and extracted with HCl (1%) to remove unreacted amine. The or-
ganic layer was washed with water and dried (MgSO4). After the sol-
vent was removed the crude material was purified on silica gel col-
umn chromatography using 2–5% methanol in methylene chloride as
eluent. The yield was 1.84 g (43%). M.p. 121–122°C. 1H NMR
(CDCl3): 1.03 (3H, s), 1.05 (3H, s), 2.56 (2H, s), 2.58–2.68 (2H, m),
2.82–2.87 (1H, q), 4.49–4.53 (1H, m), 5.14 (2H, s), 5.16 (2H, s),
5.42–7.73 (18H, m). 13C NMR (CDCl3): 27.62, 26.68, 49.94, 54.08,
57.90, 67.23, 71.55, 71.64, 119.94, 126.14, 127.69, 127.72, 127.82,
128.37, 130.59, 137.35, 138.32. HRMS (M + H) 482.2695. For
C32H36NO3 required 482.2660.
8-(Hydroxy)-5-[2-(1-phenyl-2-methylprop-2yl)-amino]-1-hydroxyethyl]
carbostyril (11a). All structural data are in agreement with those pub-
lished by Milecki et al. (1987).
2-{2-Hydroxy-2[8-hydroxy-2(1H)-quinolinonyl]-ethylamino}propane
(11b). Crude product was purified by recrystallization from ethanol-
1
ethyl acetate. The yield was 1.06 g (75%). M.p. 235°C. H NMR
(DMSO): 0.91 (3H, t), 2.42–2.61 (2H, m), 3.65–3.80 (4H, m),
5.43–5.46 (1H, m), 6.55 (1H, d), 7.02 (1H, d), 7.16 (1H, d), 8.29 (1H,
d). HRMS 263.1326 (M + H). C14H19N2O3 requires 263.1395.
1-{[2-[(1-Phenyl-2-methylprop-2-yl)amino]-1-hydroxyethyl]-3,4-bis
(hydroxy)}-benzene (14). Adduct 13 (100 mg, 0.2 mmol) was dis-
solved in 10 ml methanol. Catalyst (10% Pd/C, 80 mg) was added fol-
lowed by 1,4-cyclohexadiene (0.2 ml, 2 mmol). This mixture was re-
fluxed until the starting material was no longer detected by TLC. The
suspension was filtered through celite and then concentrated yielding
a yellow solid (33.8 mg, 56%). M.p. 102–105°C (decomposition). 1H
NMR (MeOD): 1.09 (6H, s), 2.75–2.95 (2H, m), 3.34–3.63 (2H, m),
4.51–4.60 (1H, m), 6.73–6.89 (5H, m), 7.13–7.29 (5H, m). HRMS (M
+ H) 302.1755. For C18H24NO3 required 302.1756.
2-{2-Hydroxy-2[8-hydroxy-2(1H)-quinolinonyl]-ethylamino}-2-methyl-
butane (11c). The yield was 68%. M.p. 176°C. H NMR (CDCl3 +
1
MeOD): 1.01 (3H, t), 1.38 (6H, d), 1.20–1.35 (2H, m), 3.10–3.35
(4H, m), 5.52 (1H, d), 6.65 (1H, d), 7.05 (1H, d), 7.35 (1H, d), 8.45
(1H, d). 13C NMR (CDCl3 + MeOD): 6.90, 21.52, 21.64, 23.67,
30.23, 32.65, 60.15, 65.92, 113.84, 117.55, 120.82, 128.19, 128.19,
137.38, 144.0, 162.59. HRMS 291.1679 (M + H). C16H23N2O3 re-
quires 291.1709.
5-[2-(N-Phenethyl)-1-hydroxyethylamino]-8-hydroxy-2(1H)-quinol-
inone (11d). The yield was 85%. The product was purified by column
chromatography on silica gel using 5–7% methanol in methylene
chloride as eluting system. M.p. 132–134°C. 1H NMR (MeOD):
2.80–2.98 (2H, m), 3.01–3.08 (2H, m), 3.31 (2H, M), 5.10–5.15 (1H,
m), 6.80 (1H, m), 6.90–7.0 (1H, m), 7.05–7.17 (6H, m), 8.25–8.40
(1H, m). HRMS 325.1552 (M + H). C19H21N2O3 requires 325.1551.
Biological methods
Cell culture
DDT cells were grown as monolayers on poly-L-lysine-treated 150-
mm plastic petri dishes. The dishes were prepared by adding 10 ml
sterile water containing 0.01 mg/ml poly-L-lysine and after 1 h at
room temperature they were rinsed with 2×5 ml water. Cells were
grown in 30 ml Dulbecco’s Modified Eagle’s Medium (DMEM) sup-
plemented with 5% fetal bovine serum, 100 U/ml penicillin G, 2.5
µg/ml amphotericin B, and 0.1 mg/ml streptomycin sulfate in a hu-
midified atmosphere of 95% air and 5% CO2 at 37°C. Cells were sub-
cultured twice weekly by detachment in 10 ml Hank’s Balanced Salt
Solution (HBSS) without divalent cations and containing 1 mM ED-
TA. Cells were seeded at 2–5×105 per dish and experiments per-
formed on 1 day preconfluent cultures.
5-{2-[2-(Phenylprop-2-yl)amino]-1-hydroxyethyl}-8-hydroxy-2(1H)-
quinolinone (11e). The crude product was purified on preparative
TLC using 15% methanol in methylene chloride. The yield was 170
mg (71%). M.p. 160–162°C. 1H NMR (DMSO): 1.8 (6H, d),
2.75–2.85 (2H, m), 6.50 (1H, d), 6.95 (1H, d), 7.15 (1H, d), 7.38–7.50
(3H, m), 7.60–7.65 (2H, m), 7.9 (1H, d), 8.50 (1H, m). HRMS
339.1730 (M + H). C20H23N2O3 requires 339.1708.
5-[2-(2-Methyl-5-phenylpent-2-yl)amino-1-hydroxyethyl]-8-hydroxy-
2(1H)-quinolinone (11f). Yield was 0.12 g (63%). M.p. 122–124°C.
1H NMR (DMSO) 1.23 (6H, s), 1.62 (4H, s), 2.50–2.60 (2H, m),
2.82–2.98 (2H, m), 5.40 (1H, d), 6.0–7.0 (broad signal), 6.60 (1H, d),