4
ꢀꢀꢀꢀꢁꢀA.W. Gann and P.S. Ray: A model for a potential pyrimido[4,5-e][1,4]diazepine-based folate anti-tumor agent
.79 (s, 2H), 5.26 (s, 2H), 7.14–7.28 (m, 5H); 13C NMR (CDCl ): δ 33.79, dichloromethane. The resultant solid material was chromatographed
3
3
4
1.46, 52.11, 54.95, 115.11, 116.05, 126.33, 128.45, 128.54, 138.95, 160.52, on silica gel eluting with 10% methanol in chloroform. The fractions
.
+
1
63.47. HR-MS (LIFDI-TOF). Calcd for C H Cl N , [M ] : m/z 335.0705. with an R of 0.2 were combined and the solvent removed to give 0.1 g
15 15 2 5
f
1
Found: m/z 335.0706.
(10%) of a colorless solid; mp 82–83°C; H NMR (CDCl ): δ 2.77 (d, J ꢀ=ꢀ
3
4
Hz, 2H), 2.81 (m, J ꢀ=ꢀ 8 Hz, 2H), 2.92 (t, J ꢀ=ꢀ 4 Hz, 2H), 3.13 (t, J ꢀ=ꢀ 4 Hz,
1
3
2
H), 3.54 (s, 2H), 3.86 (s, 3H), 5.05 (br s, 1H), 7.06–7.24 (m, 5H) C NMR
(
DMSO-d ): δ 31.9, 37.1, 47.5, 50.2, 54.1, 55.0, 103.4, 126.3, 128.7, 128.9,
6
2
5
-Amino-4-chloro-6-phenethyl-6,7,8,9-tetrahydro-
1
40.7, 160.6, 161.7, 169.8. HR-MS (LIFDI-TOF). Calcd for C H ClN O,
16 23 5
+
H-pyrimido[4,5-e][1,4]diazepine (3)
.
[
M ] : m/z 336.1591. Found: m/z 336.1607.
A mixture of 5 (0.45 g, 1.34 mmol), DMF (7 mL), 2 ꢁ ammonia in meth-
anol (3 mL), and Raney nickel (1 g, washed with methanol to remove
most of the water) was allowed to react under 50 psi of hydrogen in a
Parr shaker for 21 h. The reaction mixture was filtered through a pad
of celite and washed well with methanol. The solvents were removed
by rotary evaporation under reduced pressure and the residue was
partitioned between dichloromethane and water. The organic layer
was separated, dried with anhydrous sodium sulfate, filtered and the
solvent was evaporated. The residue was chromatographed on silica
gel eluting with 10% methanol in chloroform. The fractions with an
2
-Amino-4-methoxy-6-phenethyl-6,7,8,9-tetrahydro-
5
H-pyrimido[4,5-e][1,4]diazepine (15)
This compound was prepared using the initial procedure described
above for 14. The filtrate (af er the removal of the Raney nickel)
was treated with potassium carbonate (0.27 g, 2.0 mmol) and the
mixture was heated at 100°C for 4 h. The solvent was re moved by
rotary evaporation under reduced pressure and the residue was
partitioned between water (30 mL) and chloroform (30 mL). The
organic layer was washed with brine (30 mL) and dried over anhy-
drous sodium sulfate, filtered and the solvent removed by rotary
evaporation. The residue was chromatographed on silica gel elut-
R of 0.5 were combined and the solvent removed to give 0.24 g (60%)
f
1
of a colorless solid; mp 134–135°C; H NMR (CDCl ): δ 2.76–2.816 (m,
3
2
H), 2.84–2.88 (m, 2H), 3.02 (m, 2H), 3.43 (m, 2H), 4.03 (s, 2H), 4.74
br s, 2H), 5.08 (br s, 1H), 7.18–7.30 (m, 5H); 13C NMR (CDCl ): δ 34.46,
(
3
ing with 10% methanol in chloroform. The fractions with an R of
4
2.91, 50.22, 55.82, 57.02, 103.11, 126.11, 128.40, 128.71, 139.96, 160.16,
f
.
+
0.53 were combined and the solvent removed to give 0.275 g (61%)
1
60.45, 167.39. HR-MS (LIFDI-TOF). Calcd for C H ClN , [M ] : m/z
15 18 5
1
of a colorless solid; mp 119–120°C; H NMR (CDCl ): δ 2.74 (m, 2H),
3
03.1251. Found: m/z 303.1237.
3
2
.87 (m, 2H), 2.98 (m, 2H), 3.30 (m, 2H), 3.84 (s, 2H), 3.87 (s, 3H), 4.65
1
3
(
br s, 2H), 4.79 (br s, 1H), 7.19–7.30 (m, 5H); C NMR (CDCl ): δ 34.2,
3
4
3.3, 47.4, 53.6, 56.7, 57.2, 90.4, 126.0, 128.4, 128.7, 140.3, 160.6, 167.7,
.
+
2
-{N-[(2-Amino-4-chloro-6-methoxypyrimidin-5-yl)
169.2. HR-MS (LIFDI-TOF). Calcd for C H N O, [M ] : m/z 299.1746.
16 21 5
methyl]-N-phenethylamino}acetonitrile (13)
Found: m/z 299.1764.
A solution of 0.5 ꢁ sodium methoxide in methanol (16.7 mL,
8
8
.33 mmol) was added dropwise to a stirred solution of 5 (2.8 g, 2-Amino-6,7,8,9-tetrahydro-6-phenethyl-3H-
.33 mmol) in anhydrous methanol (12 mL) under nitrogen. The mix-
pyrimido[4,5-e][1,4]diazepin-4(5H)-one (2), HI Salt
ture was stirred at room temperature to 10 min and then heated at
reflux for 24 h. The solvent was removed by rotary evaporation under
reduced pressure and the residue was triturated with water and the
resultant solid was filtered under reduced pressure. This solid was
further triturated with diethyl ether, filtered and dried in a vacuum
desiccator to give 1.9 g (70%) of a colorless material; mp 158–159°C;
A mixture of 15 (0.261 g, 0.87 mmol), anhydrous DMF (5 mL) and tri-
fluoroacetic acid (0.2 mL, 2.61 mmol) was heated to 60°C and stirred
for 0.5 h. Trimethyliodosilane (0.248 mL, 1.74 mmol) was then added
and the mixture was stirred for an additional 0.5 h, quenched with
methanol (5 mL), cooled to room temperature and concentrated by
rotary evaporation under reduced pressure. The residue was tritu-
rated with sodium bicarbonate solution and the solid collected by fil-
tration at the pump was washed with ethanol and dried in a vacuum
desiccator to give 0.082 g (23%) of a colorless product; mp 186–188°C;
1
H NMR (CDCl ): δ 2.83 (d, J ꢀ=ꢀ 7.6 Hz, 2H), 2.88 (d, J ꢀ=ꢀ 8 Hz, 2H), 3.52 (s,
3
1
3
2
H), 3.68 (s, 2H), 3.90 (s, 3H), 5.07 (s, 2H), 7.18–7.27 (m, 5H); C NMR
(
CDCl ): δ 33.8, 41.7, 48.3, 54.5, 55.5, 104.5, 115.4, 126.2, 128.4, 128.6,
3
1
39.4, 160.9, 161.6, 170.1. HR-MS (LIFDI-TOF). Calcd for C H ClN O,
1
6
18
5
.
+
[
M ] : m/z 331.1200. Found: m/z 331.1220.
1
H NMR (DMSO-d ): δ 2.95 (m, 2H), 3.02 (m, 2H), 3.24 (m, 2H), 3.62 (m,
6
2
H), 4.17 (d, J ꢀ=ꢀ 13 Hz, 1H), 4.42 (d, J ꢀ=ꢀ 13 Hz, 1H), 6.49 (br s, 2H), 6.74
1
3
(
br s, 1H), 7.23–7.35 (m, 5H), 9.65 (br s, 1H), 10.30 (br s, 1H); C NMR
1
N -[(2-Amino-4-chloro-6-methoxypyrimidin-5-yl)
methyl]-N -phenethylethane-1,2-diamine (14)
(DMSO-d ): δ 30.6, 38.8, 45.7, 55.1, 78.1, 127.3, 129.1, 129.3, 137.3, 154.9,
6
.
+
. +
1
163.5, 165.9. HR-MS (LIFDI-TOF). m/z: [M ] Calcd for C H N O, [M ] :
15 19 5
m/z 285.1590. Found: m/z 285.1564.
A mixture of 13 (0.5 g, 1.5 mmol), DMF (7 mL), 2 ꢁ solution of
ammonia in methanol (3 mL), and Raney nickel (1 g, washed with
methanol to remove most of the water) was allowed to react under
Acknowledgments: We are grateful to the faculty research
grant and the student research assistant programs at the
University of West Georgia for financial support. We are
also thankful for funding from the University of West
5
0 psi of hydrogen in a Parr shaker for 21 h. The reaction mixture was
filtered through a pad of celite and washed well with DMF (5 mL).
The solvent was removed by rotary evaporation under reduced pres-
sure and the residue was triturated first with water and then with Georgia Institutional STEM Excellence (UWISE) program,
Brought to you by | Stockholms Universitet
Authenticated
Download Date | 11/28/15 6:27 PM