Synthesis of condensed quinolines and quinazolines as DNA ligands

Article abstract of DOI:10.1016/j.bmc.2003.10.014

Among new condensed quinolines and quinazolines the design of which were inspired by anti-cancer DNA-binding alkaloids such as camptothecin and batracyclin, DNA binding tests identify the 8-methoxy-7- piperazinylpropoxyindeno[1,2-b]quinolin-11-one tetracyclic system as a new motif for DNA recognition.

Full text of DOI:10.1016/j.bmc.2003.10.014

Bioorganic & Medicinal Chemistry 12 (2004) 641–647
Synthesis of condensed quinolines and quinazolines
as DNA ligands
Natacha Malecki,a Pascal Carato,a Benoıt Rigo,b Jean-Francois Goossens,a
Raymond Houssin,a Christian Baillyc and Jean-Pierre Henicharta,*
a
´
Institut de Chimie Pharmaceutique Albert Lespagnol, Universite de Lille 2, EA 2692, 3rue du Professeur Laguesse, BP 83,
F-59006 Lille, France
b
´
´
Laboratoire d’Ingenierie Moleculaire, Ecole des Hautes Etudes Industrielles, 13rue de Toul, F-59046 Lille, France
cINSERM U-524 et Laboratoire de Pharmacologie Antitumorale du Centre Oscar Lambret, Place de Verdun,
F-59045 Lille, France
Received 31 January 2003; accepted 10 October 2003
Abstract—Among new condensed quinolines and quinazolines the design of which were inspired by anti-cancer DNA-binding
alkaloids such as camptothecin and batracyclin, DNA binding tests identify the 8-methoxy-7-piperazinylpropoxyindeno[1,2-b]qui-
nolin-11-one tetracyclic system as a new motif for DNA recognition.
# 2003 Elsevier Ltd. All rights reserved.
1. Introduction
in the structure of anti-cancer tyrosine-kinase inhibitors
such as PD153035.9 We have extensively studied this
The quinoline ring is frequently condensed with various
heterocycles in the skeleton of the numerous natural
alkaloids acting as DNA ligands and potentially useful
as anti-cancer drugs. This is the case for several plant
alkaloids such as camptothecin (CPT), isolated from
Camptotheca acuminata,1 a powerful antitumor agent
targeting DNA topoisomerase I. The indolizino[1,2-
b]quinoline CPT scaffold represented by the A–D rings
provides the necessary framework for DNA interaction
whereas the lactone E-ring interacts essentially, if not
exclusively, with the active site of topoisomerase I.2 This
is also the case for mappicine ketone3 (MPK), an
analogue of mappicine isolated from Mappia foetidia
Miers, identified as an antiviral agent acting at the
DNA level.4,5 We have recently developed a strategy
which consists in eliminating the lactone E-ring of
camptothecin, therefore prohibiting the targeting of
topoisomerase I, but exploiting the indolizino[1,2-
b]quinoline structure to design DNA sequence reading
molecules.6 On the other hand, the quinazoline ring has
often been found in the structure of anti-cancer DNA-
binding agents, such as luotonin A7 or batracyclin,8 or
family of compounds10,11 and demonstrated that a
slight modification in structure (methylation of the ani-
lino group for example) could considerably reinforce the
DNA-intercalating capacities of these small molecules.12
Moreover, the observation that the presence of alkoxy
groups in ortho-position on the benzo group of quino-
line13 could provide specific cytoxicity on the human
prostate carcinoma PC3 cell line14 confirmed the
importance of such catechol groups found in other
chemical families6,10,11,15 including camptothecin
derivatives such as lurtotecan.16
Keywords: Quinolines; Quinazolines; DNA ligands.
* Corresponding author. Tel.: +33-3-2096-4374; fax: +33-3-2096-
4906; e-mail: henicha@pharma.univ-lille2.fr
0968-0896/$ - see front matter # 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2003.10.014

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N. Malecki et al. / Bioorg. Med. Chem. 12 (2004) 641–647
On the basis of these three sets of considerations, we
propose (i) the design of new condensed quinolines
Reduction of 6a,b, 7a,b as well as commercial benzal-
dehyde 6c and acetophenone 7c was selective, according
to a method already published (Fe/HCl);19 the crude
amines 8a–c and 9a–c were obtained at a very high yield
and used as such in the next step, due to their instability.
obtained by
a Friedlander-type reaction between
an aminobenzaldehyde (or aminoacetophenone) sub-
stituted by a methoxy and an alkoxy (or aminoalkoxy)
group and a cyclic ketone and (ii) the design of
dialkoxyquinazolines built from the same starting
materials (Fig. 1).
2.1.2. 5,6-Dihydrobenzo[a] or [c]acridines 11c, 13b,c,
15b,c (B). Although the synthesis of the tetramethoxy
dihydrobenzoacridine 15c has already been performed
by heating the hydrochloride of acetophenone 9c with
tetralone 14 at 140 ꢀC in a sealed vessel,20 we chose to
perform the Friedlander cyclisations (Scheme 3)
between o.carbonylanilines 9b,c and the tetralones 10,
12, 14 in acidic medium (acetic acid).
2. Results and discussion
2.1. Chemistry
2.1.1. 2-Amino- benzaldehydes 8a–c and acetophenones
9a–c (A). The synthesis of the required 2-amino-
benzaldehydes 8a–c and 2-aminoacetophenones 9a–c
was to be obtained by reducing the corresponding nitro
compounds. Whatever the reaction conditions used,
nitration of aldehyde 1 unexpectedly led to the corre-
sponding 3-nitrobenzaldehyde 2 (Scheme 1), as proven
by a HBMC proton–carbon correlation study (Fig. 2).
While a-tetralones 12 and 14 resulted in dihydro-
benzo[c]acridines 13 and 15, b-tetralone 10 gave a very
good yield of the single dihydrobenzo[a]acridine 11. The
poor yields corresponding to aminopropoxy benzo[c]a-
cridines 13b and 15b are explained by their degradation
on silica gel and required purification by chromato-
graphy on alumina.
It was therefore anticipated that substitution of the
hydroxy phenol group would induce electrophilic
substitution in ortho-position of the carbonyl function,
as previously indicated.17,18
2.1.3. 11H-indeno[1,2-b]quinolines 17b,c and their analo-
gues 19b,c, 21a–c, 22a–c (C). These fused quinolines
were obtained following the same procedure, from
o.carbonylanilines 8a–c, 9a–c and 1-indanone 16 or its
analogues 18, 20 leading to the condensed tetracycles
17, 19, 21 (21c21), 22 (Scheme 4).
Reaction of 3-dialkylaminopropyl chlorides R2Cl
(R2=a,b) with phenols 1 and 3 (Scheme 2) easily
produced aryl ethers 4a,b and 5a,b whose nitration gave
good to moderate yields of compounds 6a,b and 7a,b.
NMR correlation studies (ROESY and HBMC) on 6a
and 7a confirmed the ortho position of the nitro group
relative to the carbonyl group (Fig. 3).
As for the aminoalkoxy heterocycles 13b and 15b, their
analogues 17b, 19b, 21a,b, 22a,b were obtained at very
low yields. It is also interesting to compare the yields of the
homologous compounds 17c (37%) and 13c (71%) which
reflect the lower reactivity of indanones vs. tetralones, as
Figure 1. Design of condensed quinolines and quinazolines.
Scheme 2. Reagents and conditions: (a) R2Cl, K2CO3, DMF; (b)
HNO3 68%, T<10 ꢀC, 1 h; (c) Fe/HCl, AcOH/EtOH, reflux, 1 h.
Scheme 1. Reagents and conditions: (a) HNO3, CH2Cl2, À50 ꢀC.
Figure 3. Main ROESY correlations (a) for 6a, 7a and (b) HBMC
correlations for 6a.
Figure 2. Main HBMC correlations for phenol 2.

N. Malecki et al. / Bioorg. Med. Chem. 12 (2004) 641–647
643
Scheme 5. Reagents and conditions: (a) HCONH2, AcOH, 2 h.
Table 1. DNA binding properties
Compd
Kapp (106 MÀ1 a
)
ꢀTm (ꢀC)b
17b
21a
21b
22a
22b
PD153035
0.22Æ0.03
4.25Æ0.07
2.25Æ0.06
1.91Æ0.06
5.26Æ0.08
ndc
1.5
5.8
6.8
2.9
10.9
0.6d
a Apparent binding constant measured by fluorescence (n=3).
b Variation of the ꢀTm (Tm
alone) of the
complexÀTm
drugÀDNA
DNA
complexes between DNA and the test compounds.
c Not determined.
d From ref 12.
Scheme 3. Reagents and conditions: (a) AcOH, reflux, 2 h.
ammonia solution of formanilide in a stainless steel
bomb at 140 ꢀC (82% yield).
2.2. Biological evaluation
All compounds were evaluated for DNA binding with
two pharmacological tests using calf thymus DNA. A
fluorescence assay was applied to determine binding
constants and the results were compared to melting
temperature measurements which can give information
on the relative binding affinities of the compounds for
any given DNA. Most compounds showed little inter-
action with DNA and their affinity was too low to be
precisely calculated. In contrast, five compounds, 17b,
21a,b, 22a,b as listed in Table 1, were found to compete
with ethidium bromide for DNA binding and induced
more or less pronounced stabilization of the duplex
structure of DNA, as judged from the Tm analysis.
For clarity, only data for these five compounds with
Kapp>104 MÀ1 and ꢀTm>1 ꢀC is reported here. The
Kapp values of other compounds were between 0.05 104
MÀ1 and 0.7Â104
M
À1. A relative agreement was
observed between the two assays, Tm and fluorescence
quenching, used to monitor drug–DNA interaction.
Slight differences were observed and are probably
accounted for by varying experimental conditions
(heating vs competition). Compound 22b with a piper-
azinyl side chain was found to bind most tightly to
DNA whereas the analogue 22a with a dimethylamino
chain was 3–4 times less potent in terms of DNA bind-
ing. The effect is clearly superior to that of the reference
compounds PD153035 and CPT which both exhibit
little interaction with DNA. Surprisingly, the same
difference was not observed with the unmethylated
analogues 21a,b. Although no precise structure–DNA
binding relationships could be delineated, it was inter-
esting to observe that dialkoxyquinazolines 25a,b,c and
26a,b,c appeared to show little interaction with DNA, in
contrast to our expectation on the basis of previous
observations with related quinazoline derivatives.12
Scheme 4. Reagents and conditions: (a) AcOH, reflux, 2 h.
already observed22 for the Friedlander reaction of
indanones 23 and 24.
2.1.4. Quinazolines 25a–c, 26a–c (D). With the same
starting materials 8a–c, 9a–c and following the described
method,23 quinazolines 25 and 26 were easily obtained
by reaction with formamide (Scheme 5) whereas 26c has
previously been obtained24 by heating an ethanolic

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N. Malecki et al. / Bioorg. Med. Chem. 12 (2004) 641–647
3. Conclusion
4.1.3. 4a (hydrochloride). 61% yield; mp 83–84 ꢀC (tolu-
ene); TLC Rf [CH2Cl2/CH3OH/NH4OH(90:8:2)]=0.5;
IR (KBr) n 1680 cmÀ1; 1HNMR (CDCl 3): 2.45 (2H, m),
2.86 (6H, s), 3.29 (2H, t, J=7.8 Hz), 3.88 (3H, s), 4.22
(2H, t, J=5.7 Hz), 6.96 (1H, d, J=8.2 Hz), 7.37 (1H, d,
J=1.6 Hz), 7.42 (1H, dd, J=8.2, 1.6 Hz), 9.82 (1H, s).
The results identify the indeno[1,2-b]quinolin-11-one
tetracyclic ring system as a new motif for DNA recog-
nition and support our recent finding that the A–D
motif of the antitumor drug camptothecin offers a DNA
binding element. Synthetic efforts are now directed
towards substitution of the anilino group to increase
DNA binding capacity (and possibly to restore kinase
inhibitory activity) by analogy with previously studied
anilinoquinazolines.11
4.1.4. 5a (hydrochloride). 60% yield; mp 177–178 ꢀC
(EtOH);
TLC
Rf
[CH2Cl2/CH3OH/NH4OH
1HNMR
(90:8:2)]=0.6; IR (KBr) n 1665 cmÀ1
;
(DMSO-d6): 2.20 (2H, m), 2.53 (3H, s), 2.76 (6H, s),
3.19 (2H, t, J=7.6 Hz), 3.82 (3H, s), 4.15 (2H, t, J=6.1
Hz), 7.08 (1H, d, J=8.3 Hz), 7.45 (1H, s), 7.61 (1H, d,
J=8.3 Hz), 11.03 (1H, s, exchangeable).
4. Experimental
4.1. Chemistry
4.1.5. 5b (dihydrochloride). 59% yield; mp 217–218 ꢀC
(EtOH);
TLC
Rf
[CH2Cl2/CH3OH/NH4OH
1HNMR
(90:8:2)]=0.4; IR (KBr) n 1670 cmÀ1
;
Melting points were determined on a Buchi 535 capil-
lary melting point apparatus and remain uncorrected.
Thin layer chromatography was performed on
precoated Kieselgel 60F254 plates (Merck) and column
chromatography on silica gel 60 230-400 mesh ASTM
(Merck) or activated neutral alumina 50–160 mm
(Prolabo). The IR spectra were recorded on a Bruker
Vector 22 spectrophotometer and the NMR spectra on a
Bruker AC 300P or a Bruker DPX 300 AVANCE at 300
MHz, using tetramethylsilane as an internal reference.
Elemental analyses were performed by the Service
Central d’Analyses (CNRS, Vernaison, France).
(DMSO-d6): 2.24 (2H, m), 2.54 (3H, s), 2.84 (3H, s),
3.18–3.80 (10H, m), 3.84 (3H, s), 4.18 (2H, t, J=6.0
Hz), 7.11 (1H, d, J=8.3 Hz), 7.47 (1H, s), 7.65 (1H, d,
J=8.3 Hz), 12.05 (2H, s, exchangeable).
4.1.6. 5-Methoxy-4-[3-(4-methyl-1-piperazinyl)propoxy]-
2-nitrobenzaldehyde (6b). Compound 4b (4 g, 11 mmol)
was added portion by portion to cooled (0–5 ꢀC) HNO3
(d=1.41, 22.3 mL, 506 mmol). The mixture was stirred
for 1 h at a temperature below 10 ꢀC. H2O (50 mL), then
K2CO3 were added and the solution was extracted with
CH2Cl2. The organic phase was evaporated after drying
(MgSO4) and the residue was recrystallized from iPr2O,
leading to a 55% yield of 6b; mp 60–61 ꢀC; TLC Rf
4.1.1. 4-Hydroxy-3-methoxy-5-nitrobenzaldehyde (2). A
solution of aldehyde 1 (1 g, 6.6 mmol) in CH2Cl2 (60
mL) was cooled to À50 ꢀC; then fuming HNO3
(d=1.49, 7.7 mL, 185 mmol) was added dropwise. After
stirring at À50 ꢀC for 1 h, the reaction was quenched
onto ice. The precipitated solid was filtered, washed
with Et2O and recrystallized from AcOEt to give a 60%
yield of nitrophenol 2; mp 80–81 ꢀC; TLC Rf
[CH2Cl2/CH3OH/NH4OH(90:8:2)]=0.6; IR (KBr)
1
n
1680, 1525, 1325 cmÀ1; HNMR (CDCl ): 2.11 (2H,
m), 2.67–3.08 (13H, m), 4.01 (3H, s), 4.24 (2H, t, J=6.2
Hz), 7.42 (1H, s), 7.64 (1H, s), 10.45 (1H, s).
3
Amines 6a, 7a and 7b were prepared in a similar way.
4.1.7. 6a. 66% yield; mp 67–68 ꢀC (iPr2O); TLC Rf
[AcOEt]=0.6; IR (KBr) n 3205, 1680, 1545, 1335 cmÀ1
;
1HNMR (DMSO- d6): 3.96 (3H, s), 7.62 (1H, s), 8.11
(1H, s), 9.87 (1H, s), 11.60 (1H, exchangeable).
[CH2Cl2/CH3OH/NH4OH(90:8:2)]=0.6; IR (KBr)
n
1680, 1525, 1340 cmÀ1; 1HNMR (CDCl 3): 2.07 (2H, m,
J=6.9, 6.6 Hz), 2.26 (6H, s), 2.47 (2H, t, J=6.9 Hz),
4.01 (3H, s), 4.23 (2H, t, J=6.6 Hz), 7.41 (1H, s), 7.66
(1H, s), 10.44 (1H, s).
4.1.2. 3-Methoxy-4-[3-(4-methyl-1-piperazinyl)propoxy]
benzaldehyde dihydrochloride (4b). A stirred mixture of
phenol 1 (10 g, 66 mmol), 1-(3-chloropropyl)-4-methyl-
piperazine dihydrochloride (16.4 g, 66 mmol) and
K2CO3 (36.3 g, 264 mmol) in DMF (80 mL) was heated
at 80 ꢀC for 4 h, then at room temperature for 48 h.
H2O (80 mL) was added and the aqueous layer was
extracted with CH2Cl2. The organic phases were dried
(MgSO4) and Et2O (200 mL) was added to the residue
obtained after evaporation. The solid obtained on
addition of a saturated Et2O solution of HCl was
recrystallized from 2-propanol, giving a 55% yield of
4b; mp 197–198 ꢀC; TLC Rf [CH2Cl2/CH3OH/NH4OH
4.1.8. 7a. 57% yield; mp 90–91 ꢀC (petroleum ether);
TLC Rf [CH2Cl2/CH3OH/NH4OH(90:8:2)]=0.7; IR
(KBr) n 1700, 1520, 1330 cmÀ1; 1HNMR (CDCl 3): 2.05
(2H, m, J=7.1, 6.6 Hz), 2.26 (6H, s), 2.47 (2H, t, J=7.1
Hz), 2.50 (3H, s), 3.97 (3H, s), 4.18 (2H, t, J=6.6 Hz),
6.75 (1H, s), 7.66 (1H, s).
4.1.9. 7b. 51% yield; mp 85–86 ꢀC (petroleum ether);
TLC Rf [CH2Cl2/CH3OH/NH4OH(90:8:2)]=0.5; IR
(KBr) n 1710, 1520, 1325 cmÀ1; 1HNMR (CDCl 3): 2.01
(2H, m), 2.64–3.01 (13H, m), 3.98 (3H, s), 4.23 (2H, t,
J=6.8 Hz), 7.47 (1H, s), 7.76 (1H, s).
(90:8:2)]=0.5; IR (KBr) n 1675 cmÀ1
;
1HNMR
(DMSO-d6): 2.26 (2H, m), 2.83 (3H, s), 3.20–3.80 (10H,
m), 3.85 (3H, s), 4.20 (2H, m), 7.20 (1H, d, J=8.2 Hz),
7.42 (1H, s), 7.57 (1H, d, J=8.2 Hz), 9.86 (1H, s), 12.10
(2H, s, exchangeable).
4.1.10. 2-Amino-5-methoxy-4-[3-(4-methyl-1-piperazinyl)
propoxy]benzaldehyde (8b). Fe (1 g, 17.4 mmol), then
HCl (0.5 mL) were added to a stirred solution of nitro
compound 6b (1 g, 2.9 mmol) in a mixture of AcOH
Amines 4a, 5a, 5b were prepared in a similar way.

N. Malecki et al. / Bioorg. Med. Chem. 12 (2004) 641–647
645
(12.5 mL), EtOH(12.5 mL) and H O (6.36 mL). After
2
7.17 (1H, s), 7.25–7.43 (3H, m), 7.49 (1H, s), 8.47 (1H,
.
d, J=6.5 Hz). Anal. calcd for C27H33N3O2 2.5H2O: C,
68.04; H, 8.04; N, 8.82. Found: C, 68.26; H, 8.01; N, 8.91.
refluxing for 1 h, the mixture was filtered on Celite, H2O
(50 mL), then K2CO3 were added. The solution was
extracted with CH2Cl2 and the organic phases were
dried (MgSO4). The aminobenzaldehyde 8b thus
obtained was immediately used for the following
Friedlander reaction.
4.1.16. 15b. 11% yield; mp 192–193 ꢀC (CH2Cl2); TLC
Rf [CH2Cl2/CH3OH/NH4OH(90:8:2)]=0.5; 1HNMR
(CDCl3): 2.16 (2H, m), 2.31 (3H, s), 2.42–2.68 (13H, m),
2.92–3.10 (4H, m), 3.95–4.07 (9H, m), 4.28 (2H, t,
J=6.9 Hz), 6.76 (1H, s), 7.17 (1H, s), 7.45 (1H, s), 8.05
Amines 8a,c and 9a,b,c were prepared in a similar way.
.
(1H, s). Anal. calcd for C29H37N3O4 H2O: C, 68.35; H,
7.71; N, 8.24. Found: C, 68.76; H, 7.48; N, 8.45.
4.1.11.
9,10-Dimethoxy-7-methyl-5,6-dihydrobenzo[c]-
acridine (13c). A solution of aminoketone 9c (1 g, 5.1
mmol) in AcOH(3 mL) was added to a refluxing solu-
tion of tetralone 12 (0.76 g, 5.6 mmol) in glacial AcOH
(2 mL). The mixture was refluxed for 2 h then cooled.
The solid was collected then washed with Et2O and the
residue was chromatographed on alumina [CH2Cl2/
4.1.17. 17b. 14% yield; mp 176–178 ꢀC (CH2Cl2); TLC
Rf [CH2Cl2/CH3OH/NH4OH(90:8:2)]=0.5; 1HNMR
(CDCl3): 2.15 (2H, m), 2.31 (3H, s), 2.40–2.71 (13H, m),
3.96 (2H, s), 4.04 (3H, s), 4.30 (2H, t, J=6.9 Hz), 7.22–
8.22 (6H, m). Anal. calcd for C26H31N3O2 1.5H2O: C,
70.24; H, 7.71; N, 9.45. Found: C, 70.48; H, 7.40; N,
9.55.
.
MeOH(9.9:0.1)] and recrystallized from CH Cl2, giving
2
a 71% yield of 13c; mp 211–212 ꢀC; TLC Rf [CH2Cl2/
CH3OH(9:1)]=0.9; 1HNMR (CDCl ): 2.61 (3H, s),
3
2.98–3.09 (4H, m), 3.98 (3H, s), 4.20 (3H, s), 7.16 (1H,
s), 7.25–7.42 (3H, m), 7.48 (1H, s), 8.48 (1H, d, J=7.2
Hz). Anal. calcd for C20H19NO2: C, 78.66; H, 6.27; N,
4.59. Found: C, 78.41; H, 6.26; N, 4.76.
4.1.18. 17c. 37% yield; mp 210–211 ꢀC (CH2Cl2); TLC
Rf [CH2Cl2/CH3OH(9:1)]=0.8; 1HNMR (CDCl 3):
2.64 (3H, s), 3.87 (2H, s), 4.04 (3H, s), 4.06 (3H, s),
.
7.15–8.24 (6H, m). Anal. calcd for C19H17NO2 H2O: C,
73.77; H, 6.19; N, 4.53. Found: C, 73.81; H, 5.89; N,
4.49.
Acridine 15c was prepared in a similar way.
4.1.12. 15c. 85% yield; mp 227–228 ꢀC (CH2Cl2) (lit.20
240 C); TLC Rf [AcOEt]=0.7; HNMR (CDCl ): 2.61
3
(3H, s), 2.92–3.10 (4H, m), 3.95–4.12 (12H, m), 6.76
(1H, s), 7.17 (1H, s), 7.51 (1H, s), 8.07 (1H, s). Anal.
.
calcd for C22H23NO4 H2O: C, 68.91; H, 6.57; N, 3.65.
Found: C, 68.62; H, 6.21; N, 3.92.
4.1.19. 19b. 14% yield; mp 174–175 ꢀC (CH2Cl2); TLC
Rf [CH2Cl2/CH3OH/NH4OH(90:8:2)]=0.5; 1HNMR
(CDCl3): 2.16 (2H, m), 2.31 (3H, s), 2.40–2.88 (13H, m),
4.10 (3H, s), 4.30 (2H, t, J=6.9 Hz), 7.25–8.32 (6H, m).
Anal. calcd for C25H29N3O3 H2O: C, 68.33; H, 7.14; N,
9.60. Found: C, 68.19; H, 6.87; N, 9.97.
ꢀ
1
.
4.1.13.
8-Methoxy-7-[3-(4-methyl-1-piperazinyl)pro-
4.1.20. 19c. 27% yield; mp 204–205 ꢀC (CH2Cl2); TLC
Rf [AcOEt]=0.9; HNMR (CDCl ): 2.87 (3H, s), 4.06
3
(3H, s), 4.11 (3H, s), 7.24–8.34 (6H, m). Anal. calcd for
.
C18H15NO3 1.5H2O: C, 67.49; H, 5.66; N, 4.37. Found:
C, 67.10; H, 5.27; N, 4.09.
1
poxy]-11H-indeno[1,2-b]quinolin-11-one (21b). A solu-
tion of aminobenzaldehyde 8b (1 g, 3.2 mmol) in AcOH
(3 mL) was added to a refluxing solution of indanedione
20 (0.5 g, 3.5 mmol) in glacial AcOH(2 mL). The
mixture was refluxed for 2 h and the solvent was
evaporated. The residue was chromatographed on
alumina [CH2Cl2/MeOH(9.9:0.1)] and recrystallized
from Et2O, giving a 25% yield of 21b; mp 171–172 ꢀC;
TLC Rf [CH2Cl2/CH3OH/NH4OH(90:8:2)]=0.5; 1H
NMR (CDCl3): 2.15 (2H, m), 2.30 (3H, s), 2.44–2.68
(10H, m), 3.99 (3H, s), 4.28 (2H, t, J=6.4 Hz), 7.07–8.17
4.1.21. 7,8-Dimethoxy-11H-indeno[1,2-b]quinolin-11-one
(21c). A solution of aminobenzaldehyde 8c (1 g, 5.5
mmol) in AcOH(3 mL) was added to a refluxing
solution of indanedione 20 (0.9 g, 6.1 mmol) in AcOH
(2 mL). The mixture was refluxed for 2 h, then the
solvent was evaporated. The residue was collected and
washed with AcOEt before the solid was chromato-
graphed on alumina [CH2Cl2/MeOH(9.9:0.1)] and
recrystallized from toluene, giving a 43% yield of 21c;
.
(7H, m). Anal. calcd for C25H27N3O3 H2O: C, 68.95; H,
6.71; N, 9.65. Found: C, 68.54; H, 6.52; N, 9.41.
Quinolines 11c, 13b, 15b, 17b,c and 19b,c were prepared
in a similar way.
mp 260 ꢀC (lit.21 287 ꢀC); TLC Rf [CH2Cl2/MeOH
1
(9.9:0.1)]=0.1 (lit.21 (CHCl3) 0.24); HNMR (CDCl ):
3
4.05 (3H, s), 4.08 (3H, s), 7.11–8.21 (7H, m). Anal. calcd
for C18H13NO3: C, 74.22; H, 4.50; N, 4.81. Found: C,
74.24; H, 4.44; N, 4.72.
4.1.14. 11c. 92% yield; mp 163–164 ꢀC (cyclohexane);
TLC Rf [AcOEt/cyclohexane (1:1)]=0.2; 1HNMR
(CDCl3): 2.87 (3H, s), 2.93–3.13 (4H, m), 4.04 (3H, s),
4.05 (3H, s), 7.24–7.58 (6H, m). Anal. calcd for
C20H19NO2 H2O: C, 74.28; H, 6.55; N, 4.33. Found: C,
74.52; H, 6.41; N, 4.63.
Quinolines 21a and 22a–c were prepared in a similar
way.
.
4.1.22. 21a. 47% yield; mp 146–147 ꢀC (Et2O); TLC Rf
4.1.15. 13b. 17% yield; mp 159–160 ꢀC (CH2Cl2); TLC
Rf [CH2Cl2/CH3OH/NH4OH(90:8:2)]=0.6; 1HNMR
(CDCl3): 2.14 (2H, m), 2.32 (3H, s), 2.42–2.72 (13H, m),
2.99–3.10 (4H, m), 4.03 (3H, s), 4.28 (2H, t, J=6.9 Hz),
[CH2Cl2/CH3OH/NH4OH(90:8:2)]=0.5;
1HNMR
(CDCl3): 2.10 (2H, m, J=7.2, 6.8 Hz), 2.28 (6H, s), 2.52
(2H, t, J=7.2 Hz), 4.00 (3H, s), 4.27 (2H, t, J=6.8 Hz),
.
7.07–7.17 (7H, m). Anal. calcd for C27H22N2O3 0.5H2O:

646
N. Malecki et al. / Bioorg. Med. Chem. 12 (2004) 641–647
C, 71.14; H, 6.24; N, 7.54. Found: C, 71.65; H, 6.08; N,
7.17.
(DMSO-d6): 2.23 (2H, m), 2.75–2.85 (9H, m), 3.23 (2H,
m), 3.99 (3H, s), 4.27 (2H, m), 7.35–7.45 (2H, m), 8.91
.
(1H, s). Anal. calcd for C15H21N3O2 2C2H2O4: C, 50.11;
H, 5.53; N, 9.23. Found: C, 50.19; H, 5.99; N, 9.28.
4.1.23. 22a. 38% yield; mp 143–144 ꢀC (Et2O); TLC Rf
[CH2Cl2/CH3OH(9.9:0.1)]=0.6; 1HNMR (CDCl 3):
2.15 (2H, m, J=7.2, 6.8 Hz), 2.28 (6H, s), 2.53 (2H, t,
J=7.2 Hz), 2.96 (3H, s), 4.01 (3H, s), 4.27 (2H, t, J=6.8
Hz), 7.22–7.95 (6H, m). Anal. calcd for
4.1.30. 26b (trioxalate). 42% yield; mp 221–222 ꢀC
(MeOH); TLC Rf [CH2Cl2/MeOH(1:1)]=0.1; 1HNMR
(DMSO-d6): 2.01 (2H, m), 2.57 (3H, s), 2.60–2.69 (8H,
m), 2.84 (3H, s), 3.44 (2H, m), 3.98 (3H, s), 4.24 (2H,
m), 7.32–7.44 (2H, m), 8.90 (1H, s). Anal. calcd for
.
C23H24N2O3 H2O: C, 70.03; H, 6.64; N, 7.10. Found: C,
70.28; H, 6.55; N, 7.22.
.
C18H26N4O2 3C2H2O4: C, 48.00; H, 5.37; N, 9.33.
Found: C, 47.18; H, 6.00; N, 9.32.
4.1.24. 22b. 18% yield; mp 167–168 ꢀC (Et2O); TLC Rf
[CH2Cl2/CH3OH(9.9:0.1)]=0.6; 1HNMR (CDCl 3):
2.10 (2H, m), 2.30 (3H, s), 2.42–2.93 (13H, m), 3.99 (3H,
s), 4.26 (2H, t, J=6.4 Hz), 7.19–7.93 (6H, m). Anal.
4.1.31. 26c (oxalate). 62% yield; mp 215–216 ꢀC
(MeCN); TLC Rf [CH2Cl2/MeOH(9.9:0.1)]=0.7; 1H
NMR (DMSO-d6): 2.81 (3H, s), 3.96–4.02 (6H, m),
7.29–7.38 (2H, m), 8.90 (1H, s). Anal. calcd for
.
calcd for C26H29N3O3 H2O: C, 69.47; H, 6.95; N, 9.35.
Found: C, 70.04; H, 6.69; N, 9.02.
.
C11H12N2O2 C2H2O4: C, 53.06; H, 4.80; N, 9.52.
Found: C, 52.89; H, 4.89; N, 9.59.
4.1.25. 22c. 51% yield; mp >260 ꢀC (AcOEt); TLC Rf
[CH2Cl2/CH3OH(9.9:0.1)]=0.1; 1HNMR (CDCl 3):
2.98 (3H, s), 4.06 (3H, s), 4.07 (3H, s), 7.25–7.97 (6H,
4.2. DNA interaction studies
.
m). Anal. calcd for C19H15NO3 0.25H2O: C, 73.65; H,
5.04; N, 4.52. Found: C, 73.43; H, 4.86; N, 4.20.
Binding constants were determined by fluorescence
using a competitive displacement assay with DNA-
bound ethidium. Fluorescence data was recorded at
room temperature with a SPEX fluorometer Fluorolog.
Excitation was at 515 nm and fluorescence emission was
monitored over the range 550 to 700 nm. Experiments
were performed with an [ethidium]/[DNA] molar ratio
of 12.6:10 and a drug concentration range of 0.01–100
4.1.26. 6,7-Dimethoxyquinazoline oxalate (25c). A stir-
red mixture of aminobenzaldehyde 8c (1 g, 5.5 mmol)
and glacial AcOH(5.5 mL) in HCONH
(28 mL) was
2
refluxed for 2 h, then cooled. H2O (20 mL), then K2CO3
were added and the solution was extracted with CH2Cl2.
After drying (MgSO4), the organic phase was evapo-
rated and the residue was chromatographed on alumina
[CH2Cl2/MeOH(9.9:0.1)]. A solution of oxalic acid
(2.08 g, 16.5 mmol) in AcOEt was added dropwise to an
AcOEt solution of the crude residue resulting from
chromatography and the precipitate was washed with
Et2O, then recrystallized from MeOH, giving a 81%
yield of 25c; mp 180–181 ꢀC; TLC Rf [CH2Cl2/CH3OH
(9:1)]=0.8; 1HNMR (DMSO- d6): 3.97 (3H, s), 3.99
(3H, s), 7.36–7.49 (2H, m), 9.07 (1H, s), 9.30 (1H, s).
mM in BPE buffer at pH7.1 (6 mM Na HPO4, 2 mM
2
NaH2PO4, 1 mM EDTA). C50 values for ethidium
displacement were calculated using a fitting function
incorporated into Prism 3.0 and the apparent
equilibrium binding constants (Kapp) were calculated as
follows:
Kapp ¼ ð1:26 mM=C50Þ ꢀ Kethidium
;
.
Anal. calcd for C10H10N2O2 1.5C2H2O4: C, 48.01; H,
4.03; N, 8.61. Found: C, 47.90; H, 4.23; N, 8.25.
with Kethidium ¼ 107 Mꢁ1
:
Quinazolines 25a,b and 26a–c were prepared in a similar
way.
Melting temperature (Tm) measurements were
performed in BPE buffer using 20 mM calf thymus DNA
and 20 mM of test compound in 1 mL quartz cuvettes at
260 nm with a heating rate of 1 ꢀC/min. The Tm values
were obtained from first-derivative plots.
4.1.27. 25a (dioxalate). 65% yield; mp 221–222 ꢀC
(MeOH); TLC Rf [CH2Cl2/MeOH(1:1)]=0.1; 1HNMR
(DMSO-d6): 2.22 (2H, m), 2.80 (6H, s), 3.22 (2H, m),
3.94 (3H, s), 4.28 (2H, m), 7.32–7.50 (2H, m), 9.03 (1H,
s), 9.29 (1H, s). Anal. calcd for C14H19N3O2 2C2H2O4:
C, 48.98; H, 5.25; N, 9.52. Found: C, 48.67; H, 4.86; N,
9.33.
.
Acknowledgements
4.1.28. 25b (dioxalate). 51% yield; mp 221–222 ꢀC
(MeOH); TLC Rf [CH2Cl2/MeOH(1:1)]=0.1; 1HNMR
(DMSO-d6): 2.03 (2H, m), 2.56 (3H, s), 2.63–2.78 (8H,
m), 3.44 (2H, m), 3.96 (3H, s), 4.26 (2H, m), 7.35–7.51
(2H, m), 9.07 (1H, s), 9.30 (1H, s). Anal. calcd for
This work was completed with the support of research
grants (to C.B. and J.-P.H.) from the Association pour
la Recherche sur le Cancer.
.
C17H24N4O2 2.5C2H2O4: C, 48.80; H, 5.40; N, 10.35.
Found: C, 47.97; H, 5.84; N, 10.00.
References and notes
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