K. Yokoyama et al. / Bioorg. Med. Chem. 16 (2008) 7968–7974
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7.1.11. 2-[(1-{4-[(4-Chlorophenyl)amino]-6,7-dimethoxyquinaz-
olin-2-yl}piperidin-4-yl)(methyl)amino] ethanol (8e)
m), 3.93 (3H, s), 3.95 (3H, s), 7.51–7.62 (2H, m), 7.72–7.79 (3H, m),
8.16 (1H, br), 10.80 (1H, br), 10.96 (1H, br), 11.61 (1H, br).
Anal. Calcd for C27H37N6ClO2 Á3HClÁ4H2O: C, 46.69; H, 6.97; N,
12.10; Cl, 20.42. Found: C, 46.64; H, 6.84; N, 11.95; Cl, 20.93.
The hydrochloride salt of 8e (109 mg, 19%) was obtained as a
pale yellow solid from crude 7c (386 mg), 2-[methyl(piperidin-4-
yl)amino]ethanol 5a (347 mg) and DBU (0.45 mL) using procedures
similar to those described for the synthesis of 8d.
7.1.16. N-(4-Chlorophenyl)-2-(1,4-diazepan-1-yl)-6,7-
dimethoxyquinazolin-4-amine (9)
MS (FAB+) m/z 472 [M+H]+. 1H NMR (400 MHz, DMSO-d6) d 1.67–
1.82 (2H, m), 2.05–2.24 (2H, m), 2.73 (3H, d, J = 4.8 Hz), 2.98–3.22
(3H, m), 3.23–3.42 (2H, m), 3.72–3.81 (2H, m), 3.92 (3H, s), 3.94
(3H, s), 4.66 (2H, br), 7.54 (2H, d, J = 8.8 Hz), 7.68–7.76 (3H, m),
8.14 (1H, br), 10.05 (1H, s), 10.96 (1H, s), 12.75 (1H, s).
Compound 8j (3.86 g, 70%) was obtained as a pale yellow solid
from crude 7c (3.77 g), tert-butyl 1,4-diazepane-1-carboxylate
(2.38 g, 11.9 mmol) and DBU (3.5 mL) using procedures similar to
those described for the synthesis of 8d. 4 M-HCl dioxane solution
(7.5 mL) was added to a solution of 8j (3.86 g) in dioxane
(50 mL), and the resulting mixture was stirred at room tempera-
ture overnight. The reaction mixture was concentrated in vacuo,
and the residue was recrystallized from EtOH-Et2O to yield N-(4-
chlorophenyl)-2-(1,4-diazepan-1-yl)-6,7-dimethoxyquinazolin-4-
amine 9 (3.76 g, quant.) as a pale yellow solid.
Anal. Calcd for C24H30ClN5O3 Á2HClÁ2.3H2O: C, 49.16; H, 6.29; N,
11.94; Cl, 18.14. Found: C, 49.04; H, 6.03; N, 11.98; Cl, 18.22.
7.1.12. 3-[(1-{4-[(4-Chlorophenyl)amino]-6,7-dimethoxyquinaz-
olin-2-yl}piperidin-4-yl)(methyl)amino]propan-1-ol (8f)
The hydrochloride salt of 8f (175 mg, 30%) was obtained as a
pale yellow solid from crude 7c (386 mg), 3-[methyl(piperidin-4-
yl)amino]propan-1-ol 5b (368 mg, 1.5 mmol) and DBU (0.45 mL)
using procedures similar to those described for the synthesis of 8d.
MS (ESI+) m/z 486 [M+H]+. 1H NMR (400 MHz, DMSO-d6) d 1.60–
1.95 (4H, m), 2.08–2.25 (2H, m), 2.68 (3H, d, J = 4.8 Hz), 2.95–3.25
(3H, m), 3.30–3.52 (2H, m), 3.55–3.67 (2H, m), 3.91 (3H, s), 3.94
(3H, s), 4.68–4.84 (2H, m), 7.53 (2H, d, J = 8.8 Hz), 7.70–7.78 (3H,
m), 8.18 (1H, br), 10.64 (1H, s), 11.04 (1H, s), 12.84 (1H, s).
Anal. Calcd for C25H32ClN5O3 Á2HClÁ1.8H2O: C, 50.78; H, 6.41; N,
11.84; Cl, 17.99. Found: C, 50.87; H, 6.12; N, 11.96; Cl, 18.02.
MS (ESI+) m/z 496 [M+H]+. 1H NMR (400 MHz, DMSO-d6) d
2.00–2.20 (2H, m), 3.50–3.65 (8H, m), 3.88 (3H, s), 3.91 (3H, s),
7.45–7.57 (2H, m), 7.74–7.86 (3H, m), 8.22 (1H, s).
7.1.17. N-(4-Chlorophenyl)-6,7-dimethoxy-2-[4-(2-pyrrolidin-
1-ylethyl)-1,4-diazepan-1-yl]quinazolin-4-amine (8k)
1-(2-Chloroethyl)pyrrolidine hydrochloride (255 mg, 1.5 mmol)
was added to a solution of 9 (487 mg, 1 mmol) in DMF (10 mL), so-
dium carbonate (318 mg), and sodium iodide (150 mg). The result-
ing mixture was stirred at room temperature for 16 h. The reaction
mixture was concentrated in vacuo. Water was added to the residue,
and the mixture was extracted with chloroform. The organic layer
was washed with brine, and then dried over sodium sulfate, filtered,
and evaporated in vacuo. The residue was purified using column
chromatography (chloroform/MeOH) to yield 8k. Compound 8k
was treated with 4M–HCl ethyl acetate solution, and the resulting
precipitate was recrystallized from an EtOH–Et2O-H2O mixture to
yield the hydrochloride salt of 8k (248 mg, 36% from 9) as a pale yel-
low solid.
7.1.13. 4-[(1-{4-[(4-Chlorophenyl)amino]-6,7-dimethoxyquinaz-
olin-2-yl}piperidin-4-yl)(methyl)amino]butan-1-ol (8g)
The hydrochloride salt of 8g (107 mg, 18%) was obtained as a
pale yellow solid from crude 7c (387 mg), 4-[methyl(piperidin-4-
yl)amino]butan-1-ol 5c (389 mg, 1.5 mmol) and DBU (0.45 mL)
using procedures similar to those described for the synthesis of 8d.
MS (FAB+) m/z 500 [M+H]+. 1H NMR (400 MHz, DMSO-d6) d
1.38–1.52 (2H, m), 1.68–1.84 (4H, m), 2.08–2.24 (2H, m), 2.66
(3H, d, J = 4.8 Hz), 2.90–3.04 (1H, m), 3.06–3.22 (2H, m), 3.28–
3.64 (4H, m), 3.90 (3H, s), 3.94 (3H, s), 4.69–4.84 (2H, m), 7.53
(2H, d, J = 8.8 Hz), 7.72–7.82 (3H, m), 8.19 (1H, br), 10.65 (1H, s),
11.06 (1H, s), 12.87 (1H, s).
MS (ESI+) m/z 511 [M+H]+. 1H NMR (400 MHz, DMSO-d6) d 1.03–
1.12 (2H, m), 1.80–2.08 (4H, m), 2.97–3.12 (2H, m), 3.20–3.28 (2H,
m), 3.38–3.50 (8H, m), 3.54–3.76 (4H, m), 3.91 (3H, s), 3.95 (3H, s),
7.49–7.60 (2H, m), 7.74–7.84 (3H, m), 8.20–8.30 (1H, m), 9.39 (1H,
s), 11.03–11.25 (2H, m).
Anal. Calcd for C26H34ClN5O3 Á1.9HClÁ0. 4H2O: C, 52.44; H, 6.21;
N, 11.76; Cl, 17.26. Found: C, 52.41; H, 6.52; N, 11.88; Cl, 17.42.
Anal. Calcd for C27H35N6ClO2 Á3HClÁ4.1H2O: C, 46.71; H, 6.71; N,
12.10; Cl, 20.42. Found: C, 46.69; H, 6.24; N, 12.31; Cl, 20.21.
7.1.14. N-(4-Chlorophenyl)-2-(4-cyclohexyl-1,4-diazepan-1-yl)-
6,7-dimethoxyquinazolin-4-amine (8h)
The hydrochloride salt of 8h (44 mg, 7%) was obtained as a pale
yellow solid from crude 7c (386 mg), 1-cyclohexyl-1,4-diazepane
(1.5 mmol) and DBU (0.45 mL) using procedures similar to those
described for the synthesis of 8d.
7.1.18. N-(4-Chlorophenyl)-6,7-dimethoxy-2-[4-(2-piperidin-1-
ylethyl)-1,4-diazepan-1-yl]quinazolin-4-amine (8l)
The hydrochloride salt 8l (252 mg, 35%) was obtained as a pale
yellow solid from 9 (487 mg, 1 mmol), 1-(2-chloroethyl)piperidine
hydrochloride (276 mg, 1.5 mmol) and sodium iodide (150 mg)
using procedures similar to those described for the synthesis of 8k.
MS (ESI+) m/z 525 [M+H]+. 1H NMR (400 MHz, DMSO-d6) d 1.35–
1.46 (2H, m), 1.75–1.85 (4H, m), 2.25–2.35 (2H, m), 2.87–3.01 (2H,
m), 3.30–3.49 (8H, m), 3.51–3.85 (6H, m), 3.91 (3H, s), 3.95 (3H, s),
7.50–7.62 (2H, m), 7.73–7.85 (3H, m), 8.26 (1H, s, br), 10.66 (1H,
br), 11.15 (1H, br), 11.48 (1H, br).
MS (ESI+) m/z 496 [M+H]+. 1H NMR (400 MHz, DMSO-d6) d 1.02–
1.18 (2H, m), 1.20–1.50 (4H, m), 1.54–1.65 (1H, m), 1.74–1.84 (2H,
m), 2.02–2.14 (3H, m), 3.14–3.34 (4H, m), 3.42–3.57 (2H, m), 3.68–
3.85 (2H, m), 3.92 (3H, s), 3.94 (3H, s), 4.18–4.36 (1H, m), 7.46–
7.60 (2H, m), 7.68–7.82 (3H, m), 8.18 (1H, s, br), 10.79 (1H, s),
10.99 (1H, s).
Anal. Calcd for C27H34N5ClO2 Á1.9HClÁ3H2O: C, 52.36; H, 6.82; N,
Anal. Calcd for C28H37N6ClO2 Á3HClÁ4.2H2O: C, 47.36; H, 6.87; N,
11.31; Cl, 16.60. Found: C, 52.35; H, 6.73; N, 11.70; Cl, 16.63.
11.83; Cl, 19.97. Found: C, 47.20; H, 6.52; N, 11.79; Cl, 19.71.
7.1.15. N-(4-Chlorophenyl)-2-{4-[2-(diethylamino)ethyl]-1,4-
diazepan-1-yl}-6,7-dimethoxyquinazolin-4-amine (8i)
7.2. Pharmacology
The hydrochloride salt of 8i (170 mg, 24%) was obtained as a
pale yellow solid from crude 7c (386 mg), 2-(1,4-diazepan-1-yl)-
N,N-diethylethanamine (463 mg, 1.5 mmol) and DBU (0.45 mL)
using procedures similar to those described for the synthesis of 8d.
MS (ESI+) m/z 513 [M+H]+. 1H NMR (400 MHz, DMSO-d6) d 1.20–
1.28 (6H, m), 3.12–3.23 (4H, m), 3.34–3.39 (6H, m), 3.52–3.63 (6H,
7.2.1. Human and murine CCR4-expressing cells
Cells from the mouse pre-B cell line B300-19 were cultured in
RPMI 1640 medium containing 10% fetal bovine serum (FBS),
50 lM 2-mercaptethanol, 100 U/mL penicillin, and 100 lg/mL
streptomycin. The expression vector pEF-BOS-Neo,17 carrying
full-length human CCR4 cDNA (X85740; GenBank) or mouse