9306 J. Am. Chem. Soc., Vol. 123, No. 38, 2001
Ellis et al.
mg, 6.4 µmol, 1 equiv) and EDCI (1.8 mg, 9.6 µmol, 1.5 equiv). The
reaction mixture was stirred for 12 h at 25 °C before the solvent was
removed under a stream of N2. Chromatography (0.7 cm × 6 cm, SiO2,
30:1 CHCl3/CH3OH) afforded 14 (3.9 mg, 80%) as a light yellow
film: 1H NMR (DMF-d7, 400 MHz) δ 11.90 (s, 1H), 11.86 (s, 1H),
10.61 (s, 1H) 10.36 (s, 1H), 9.29 (d, J ) 2.0 Hz, 1H), 9.02 (d, J ) 1.7
Hz, 1H), 8.35 (m, 2H), 7.76 (dd, J ) 2.0, 8.9 Hz, 1H), 7.67 (m, 2H),
7.64 (d, J ) 1.7 Hz, 1H), 7.61 (s, 1H), 7.54 (m, 2H), 7.45 (t, J ) 7.2
Hz, 2H), 7.40 (t, J ) 7.5 Hz, 1H), 7.22 (d, J ) 1.4 Hz, 1H), 5.43 (s,
2H), 4.27 (m, 4H), 4.02 (dd, J ) 6.8, 11.0 Hz, 1H), 3.92 (s, 3H), 1.61
(s, 9H); IR (film) νmax 3324, 2927, 1694, 1645 cm-1; MALDI-HRMS
(DHB) m/z 799.2663 (M + H+, C44H39ClN6O7 requires m/z 799.2647).
(+)-(1S)-14: [R]25D +22 (c 0.06, acetone). ent-(-)-(1R)-14: [R]25D -20
(c 0.055, acetone).
[3,2-e]indole-8-carboxamide (17). A sample of 14 (3.5 mg, 4.4 µmol)
was treated with 200 µL of 4 N HCl/EtOAc. The yellow reaction
mixture was stirred at 25 °C for 30 min and was concentrated under a
stream of N2. The red residue was dissolved in 100 µL of anhydrous
DMF and treated sequentially with NaHCO3 (1.1 mg, 13.1 µmol) and
acetyl chloride (0.82 mg, 8.8 µmol). The reaction mixture was stirred
for 12 h and diluted with H2O. The resulting precipitate was collected
by centrifugation, washed with H2O, and dried under reduced pressure
to give 17 (2.6 mg, 81%) as a yellow film: 1H NMR (DMF-d7, 400
MHz) δ 11.89 (d, J ) 2.0 Hz, 1H), 11.86 (d, J ) 1.2 Hz, 1H), 10.61
(s, 1H), 9.30 (d, J ) 2.0 Hz, 1H), 9.03 (d, J ) 2.0 Hz, 1H), 8.51 (s,
1H), 8.33 (m, 2H), 7.76 (dd, J ) 2.1, 8.8 Hz, 1H), 7.68 (m, 2H), 7.63
(d, J ) 1.6 Hz, 1H), 7.59 (d, J ) 8.8 Hz, 1H), 7.54 (m, 2H), 7.45
(apparent t, J ) 7.1 Hz, 2H), 7.38 (apparent t, J ) 7.5 Hz, 1H), 7.20
(d, J ) 2.1 Hz, 1H), 5.41 (s, 2H), 4.52 (m, 1H), 4.37 (m, 1H), 4.24
(m, 2H), 4.04 (m, 1H), 3.90 (s, 3H), 2.33 (s, 3H); IR (film) νmax 3283,
2919, 1707, 1654, 1642 cm-1; MALDI-HRMS (DHB) m/z 741.2213
N-(2-(N-((2-Methoxycarbonyl)indol-5-yl)carbamoyl)indol-5-yl)-
1-(chloromethyl)-5-hydroxy-3-(tert-butyloxycarbonyl)-1,2-dihydro-
3H-pyrido[3,2-e]indole-8-carboxamide (15). A suspension of 14 (3.2
mg, 4.0 µmol) and 10% Pd-C (3.2 mg, 1 wt equiv) in 80 µL of
degassed, anhydrous THF was treated with 25% aq HCO2NH4 (6.3
mg, 25 µL, 100 µmol, 25 equiv). The reaction mixture was stirred for
3 h and filtered through a small plug of Celite. The filtrate was
concentrated under a stream of N2 to afford 15 (2.7 mg, 96%) as a
light yellow film: 1H NMR (DMF-d7, 250 MHz) δ 11.91 (d, J ) 2.1
Hz, 1H), 11.80 (s, 1H), 10.57 (s, 1H), 10.31 (s, 1H), 10.21 (br s, 1H),
9.22 (d, J ) 2.1 Hz, 1H), 9.01 (d, J ) 2.1 Hz, 1H), 8.35 (m, 2H), 8.04
(br s, 1H, obscured by DMF), 7.78 (dd, J ) 1.6, 8.9 Hz, 1H), 7.63 (m,
2H), 7.56 (m, 2H), 7.23 (d, J ) 2.1 Hz, 1H), 4.24 (m, 5H), 3.92 (s,
(M + H+, C41H33ClN6O6 requires m/z 741.2223). (+)-(1S)-17: [R]25
D
+213 (c 0.07, DMF). ent-(-)-(1R)-17: [R]25 -210 (c 0.07, DMF).
D
N-(2-(N-((2-Methoxycarbonyl)indol-5-yl)carbamoyl)indol-5-yl)-
3-acetyl-1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-pyrido[3,2-e]-
indole-8-carboxamide (18). A suspension of 17 (3.0 mg, 4.0 µmol)
and 10% Pd-C (3.0 mg, 1 wt equiv) in 200 µL of anhydrous THF
was treated with 25% aq HCO2NH4 (12.8 mg, 51 µL, 200 µmol, 50
equiv). The reaction mixture was stirred for 5 h and filtered through
Celite. The filtrate was concentrated under a stream of N2 to afford 18
(2.6 mg, 100%) as a light yellow residue: 1H NMR (DMF-d7, 400
MHz) δ 11.88 (s, 1H), 11.60 (s, 1H), 10.23 (s, 1H), 9.94 (s, 1H), 9.05
(s, 1H), 8.32 (d, J ) 2.1 Hz, 2H), 8.18 (d, J ) 1.7 Hz, 1H), 7.73 (m,
2H), 7.54 (m, 3H), 7.48 (s, 1H), 7.45 (m, 1H), 7.20 (d, J ) 1.7 Hz,
1H), 4.21 (m, 1H), 4.10 (dd, J ) 1.0, 9.2 Hz, 1H), 3.95 (m, 1H), 3.90
(s, 3H), 3.72 (m, 2H), 2.18 (s, 3H); IR (film) νmax 3260, 2919, 1695,
1637, 1537 cm-1; MALDI-HRMS (DHB) m/z 651.1772 (M + H+,
C34H27ClN6O6 requires m/z 651.1753). (-)-(1S)-18: [R]25D -11 (c 0.14,
3H), 1.60 (s, 9H); IR (film) νmax 3271, 2916, 2848, 1712, 1644 cm-1
;
MALDI-HRMS (DHB) m/z 709.2189 (M + H+, C37H33ClN6O7 requires
m/z 709.2177). (-)-(1S)-15: [R]25 -110 (c 0.009, DMF). ent-(+)-
D
(1R)-15: [R]25 +120 (c 0.009, DMF).
D
N-(2-(N-((2-Methoxycarbonyl)indol-5-yl)carbamoyl)indol-5-yl)-
3-(tert-butyloxycarbonyl)-1,2,9,9a-tetrahydrocyclopropa[c]pyrido-
[3,2-e]indol-4-one-7-carboxamide (Indole2-CPyI-BOC, 9). A solution
of 15 (1.3 mg, 1.8 µmol) in 100 µL of anhydrous DMF under Ar was
treated with DBU (0.82 mg, 5.4 µmol, 3 equiv). The reaction mixture
was stirred for 3 h at 25 °C before the reaction mixture was directly
subjected to flash chromatography (0.7 cm × 6 cm, SiO2, 20% CH2-
Cl2-DMF) to afford 9 (0.8 mg, 67%) as a light yellow film: 1H NMR
(DMF-d7, 250 MHz) δ 11.91 (s, 1H), 11.81 (s, 1H), 10.51 (s, 1H),
10.32 (s, 1H), 9.26 (d, J ) 1.6 Hz, 1H), 8.34 (s, 1H), 8.32 (s, 1H),
8.27 (d, J ) 2.1 Hz, 1H), 7.74 (dd, J ) 1.6, 9 Hz, 1H), 7.61 (br s, 2H),
7.55 (m, 2H), 7.22 (d, J ) 2.1, 1H), 6.92 (s, 1H), 4.22 (dd, J ) 1.6,
5.8 Hz, 1H), 4.41 (m, 1H), 3.92 (s, 3H), 3.36 (m, 1H), 2.03 (dd, J )
3.7, 7.9 Hz, 1H), 1.73 (apparent t, J ) 4.2 Hz, 1H), 1.56 (s, 9H); IR
(film) νmax 3295, 2919, 1713, 1637, 1542 cm-1; MALDI-HRMS (DHB)
m/z 695.2242 (M + Na+, C37H32N6O7 requires m/z 695.2225). (+)-
DMF). ent-(+)-(1R)-18: [R]25 +9 (c 0.14, DMF).
D
N-(2-(N-((2-methoxycarbonyl)indol-5-yl)carbamoyl)indol-5-yl)-
3-acetyl-1,2,9,9a-tetrahydrocyclopropa[c]pyrido[3,2-e]-indol-4-one-
7-carboxylamide (Indole2-CPyI-Ac, 19). A sample of 18 (1.4 mg,
2.2 mmol) was suspended in 300 µL of THF and treated with 300 µL
of saturated aq NaHCO3. The biphasic solution was stirred for 4 h,
diluted with H2O, and extracted with EtOAc. The combined organic
phases were concentrated under a stream of N2 and placed under
vacuum to afford 19 (1.2 mg, 92%) as a tan residue: 1H NMR (DMF-
d7, 400 MHz) δ 11.89 (br s, 2H), 10.52 (br s, 1H), 10.43 (br s, 1H),
9.25 (d, J ) 1.7 Hz, 1H), 8.34 (d, J ) 1.7 Hz, 2H), 8.28 (d, J ) 2.1
Hz, 1H), 7.76 (s, 1H), 7.74 (d, J ) 2.1 Hz, 1H), 7.60 (s, 2H), 7.54 (s,
1H), 7.51 (d, J ) 6.7 Hz, 1H), 7.20 (s, 1H), 4.31 (m, 2H), 3.90 (s,
3H), 3.38 (m, 1H), 2.30 (s, 3H), 2.02 (dd, J ) 3.8, 7.9 Hz, 1H), 1.72
(apparent t, J ) 4.6 Hz, 1H); IR (film) νmax 2919, 1701, 1637, 1537,
1231 cm-1; MALDI-HRMS (DHB) m/z 615.1991 (M + H+, C34H26N6O6
requires m/z 615.1991). (+)-(8bR, 9aS)-19: [R]25D +18 (c 0.06, DMF).
(8bR, 9aS)-9: [R]25D +30 (c 0.04, DMF). ent-(-)-(8bS, 9aR)-9: [R]25
-30 (c 0.04, DMF).
D
N-(2-(N-((2-Methoxycarbonyl)indol-5-yl)carbamoyl)indol-5-yl)-
1,2,9,9a-tetrahydrocyclopropa[c]pyrido[3,2-e]indol-4-one-7-carbox-
amide (Indole2-CPyI-H, 16). A sample of 15 (2.0 mg, 2.8 µmol) was
treated with 150 µL of 4 N HCl/EtOAc. The yellow reaction mixture
was stirred at 25 °C for 30 min and was concentrated under a stream
of N2. The resulting red residue was suspended in 100 µL of THF and
treated with 100 µL of saturated aq NaHCO3. The biphasic solution
was stirred for 4 h, diluted with H2O, and extracted with EtOAc. The
combined organic phases were concentrated under a stream of N2 and
placed under vacuum to afford 16 (1.3 mg, 81%) as a tan residue: 1H
NMR (DMF-d7, 400 MHz) δ 11.88 (br s, 2H), 10.41 (m, 2H), 9.18 (d,
J ) 2.1 Hz, 1H), 8.34 (d, J ) 2.1 Hz, 1H), 8.30 (s, 1H), 8.15 (d, J )
2.1 Hz, 1H), 7.74 (dd, J ) 8.8, 2.1 Hz, 1H), 7.59 (m, 2H), 7.54 (s,
1H), 7.52 (s, 1H), 7.50 (s, 1H), 7.20 (s, 1H), 5.72 (d, J ) 1.2 Hz, 1H),
3.90 (s, 4H), 3.70 (d, J ) 10.4 Hz, 1H), 3.31 (m, 1H), 1.87 (dd, J )
7.1, 3.3 Hz, 1H), 1.41 (apparent t, J ) 4.2 Hz, 1H); IR (film) νmax
3261, 2919, 1702, 1634, 1538 cm-1; MALDI-HRMS (DHB) m/z
573.1881 (M + H+, C32H24N6O5 requires m/z 573.1870). (+)-(8bR,
9aS)-16: [R]25 +14 (c 0.065, DMF). ent-(-)-(8bS, 9aR)-16: [R]25
ent-(-)-(8bS, 9aR)-19: [R]25 -20 (c 0.05, DMF).
D
Acknowledgment. We gratefully acknowledge the financial
support of the National Institutes of Health (Grant CA41986),
The Skaggs Institute for Chemical Biology, and Corixa Phar-
maceuticals. S.E.W is a Skaggs Fellow and recipient of an
ARCS fellowship. We thank Michael P. Hedrick for conducting
the cytotoxic assays.
Supporting Information Available: Experimental details
for the DNA alkylation studies, gel figures of (+)-10, ent-
(-)-10, and ent-(-)-9 in w836 DNA (Figure S1), gel figures
of (+)-1, ent-(-)-1, and (+)-9, 16, and 19 in w794 DNA (Figure
S2), a representative gel figure used to establish rates of DNA
alkylation (Figure S3), a Table S1 of absolute rate constants,
and representative data treatments for establishing rates (Figure
S4) (pdf). This material is available free of charge via the
D
D
-17 (c 0.065, DMF).
N-(2-(N-((2-Methoxycarbonyl)indol-5-yl)carbamoyl)indol-5-yl)-
3-acetyl-5-(benzyloxy)-1-(chloromethyl)-1,2-dihydro-3H-pyrido
JA010769R