Synthesis of 7-oxo-7H-naphtho[1,2,3-de]quinoline derivatives as potential anticancer agents active on multidrug resistant cell lines

Article abstract of DOI:10.1016/j.bmc.2006.01.008

Following our earlier finding that tetracyclic anthraquinone analogs with a fused pyridone ring exhibit cytotoxic activity toward multidrug resistant tumor cells, a series of new potential antitumor agents, 7-oxo-7H-naphtho[1,2,3-de] quinoline derivatives (3, 6-8, 10-12, 14, 15, and 18), bearing one or two basic side chains and various substituents at the pyridone ring, have been synthesized. The compounds have been obtained from 1-amino-4-chloroanthraquinone or 1-aminoanthraquinone by cyclization with diethyl malonate and the subsequent reactions of the key intermediates 2, 4, and 17. The compounds exhibited cytotoxic activity toward sensitive human leukemia cell line HL-60 and against its resistant sublines HL-60/VINC (MDR1 type) and HL-60/DX (MRP1 type).

Full text of DOI:10.1016/j.bmc.2006.01.008

Bioorganic & Medicinal Chemistry 14 (2006) 2880–2886
Synthesis of 7-oxo-7H-naphtho[1,2,3-de]quinoline derivatives
as potential anticancer agents active on multidrug resistant cell lines
a
a
a
b
´
Maria Dzieduszycka, Maria M. Bontemps-Gracz, Barbara Stefanska, Sante Martelli,
Agnieszka Piwkowska,a Małgorzata Arciemiuka and Edward Borowskia,*
aDepartment of Pharmaceutical Technology and Biochemistry, Gdan´sk University of Technology, 80-952 Gdan´sk, Poland
bDepartment of Chemistry, University of Camerino, 62033 Camerino, Italy
Received 4 January 2005; accepted 20 May 2005
Available online 2 February 2006
Abstract—Following our earlier finding that tetracyclic anthraquinone analogs with a fused pyridone ring exhibit cytotoxic activity
toward multidrug resistant tumor cells, a series of new potential antitumor agents, 7-oxo-7H-naphtho[1,2,3-de]quinoline derivatives
(3, 6–8, 10–12, 14, 15, and 18), bearing one or two basic side chains and various substituents at the pyridone ring, have been syn-
thesized. The compounds have been obtained from 1-amino-4-chloroanthraquinone or 1-aminoanthraquinone by cyclization with
diethyl malonate and the subsequent reactions of the key intermediates 2, 4, and 17. The compounds exhibited cytotoxic activity
toward sensitive human leukemia cell line HL-60 and against its resistant sublines HL-60/VINC (MDR1 type) and HL-60/DX
(MRP1 type).
Ó 2006 Elsevier Ltd. All rights reserved.
1. Introduction
toward resistant tumor cell lines has been shown by
the following tetracyclic anthracenedione and acridine
agents: anthrapyrazoles5,6 and their aza-analogs,7
benzoperimidines,3 anthrapyridazones,8 anthrapyri-
dones,9–11 pyrimidoacridines,12,13 pyrazoloacridines,14,15
and pyrazolopyrimidoacridines.14,16
The appearance of cancer cell populations resistant to
multi-based chemotherapy constitutes a major problem
to achieve cures in the patients. The multidrug resistance
is associated with overexpression of plasma membrane
drug efflux pumps such as P-gp (MDR1) and multidrug
resistance-associated protein (MRP1).1,2 One of the
strategies to avoid or decrease this effect is the design
and synthesis of non-cross-resistant analogs of antitu-
mor agents on a rational basis. The identification of
the structural factors of the compounds responsible for
their ability to overcome multidrug resistance is essential
for this strategy.
In the continuous search for optimized compounds, we
have synthesized a series of new 7-oxo-7H-naph-
tho[1,2,3-de]quinoline derivatives (hereafter referred to
as anthrapyridones) and studied their ability to over-
come multidrug resistance. Our earlier studies with
anthrapyridones9 comprised rather limited number of
compounds. The new derivatives presented in this paper
should allow better establishment of structural require-
ments for the ability to overcome multidrug resistance
of tumor cells by this novel group of antitumor agents.
The structures of the synthesized compounds, shown
in Table 1, include compounds with side chain attached
to the chromophore at position 6 (10–12) and to 1-pyri-
done ring (6–8 and 3), the analogs of 10–12, namely 2-Cl
compounds (14 and 15), as well as 1-carboxamide com-
pound 18. Such selection of compounds enables us to
examine the influence of the kind of functional groups
attached to the pyridone ring, as well as of the structure
of aminoalkylamino side arms at position 6, on the cyto-
toxicity of anthrapyridones and their ability to over-
come multidrug resistance.
We have earlier put forward the hypothesis that the pres-
ence of five- or six-membered heterocyclic ring(s) fused
with the anthraquinone or acridine moiety is essential
for the ability to overcome multidrug resistance by antitu-
mor compounds of these groups.3,4 The cytotoxic activity
Abbreviations: MIT, mitoxantrone; DX, doxorubicin; VINC, vincris-
tine; MDR1, P-gp dependent multidrug resistance; MRP1, multidrug
resistance associated protein dependent resistance.
Keywords: Antitumor compounds; Anthracenedione analogs; Anthra-
pyridones; Cytotoxic activity; Multidrug resistance.
*
Corresponding author. Tel.: +48 58 347 2523; fax: +48 58 347
1144; e-mail: borowski@altis.chem.pg.gda.pl
0968-0896/$ - see front matter Ó 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2006.01.008

M. Dzieduszycka et al. / Bioorg. Med. Chem. 14 (2006) 2880–2886
Table 1. Structures and characteristics of anthrapyridone derivatives
2881
Y
X
2
2
1
R2
R2
1
NH
N
6
6
O
O
R1
R1
Compound
R1
R2
COOC2H5
CONH(CH2)2N(CH3)2
X
Y
Mp (°C)
Formula
3ÆHCl
NH(CH2)2N(CH3)2
NH(CH2)2N(CH3)2
NH(CH2)2N(C2H5)2
NH(CH2)2N-c-(CH2)5
NH(CH2)2N(CH3)2
NH(CH2)2N-c-(CH2)5
NH(CH2)3N(CH3)2
NH(CH2)2N(CH3)2
NH(CH2)2N-c-(CH2)5
H
OH
OH
OH
OH
253–254
296
C23H24N3O4Cl
C25H31N5O3Cl2
C27H35N5O3Cl2
C28H35N5O3Cl2
C20H20N3O2Cl
C23H24N3O2Cl
C21H22N3O2Cl
C20H19N3OCl2
C23H23N3OCl2
C21H20N3O3Cl
6Æ2HCl
7Æ2HCl
8Æ2HCl
10ÆHCl
11ÆHCl
12ÆHCl
14ÆHCl
15ÆHCl
18ÆHCl
CONH(CH2)2N(CH3)2
CONH(CH2)2N(CH3)2
266–268
115–117
230–232
225–227
220–222
260–261
>320
H
O
O
O
H
H
H
Cl
Cl
H
CONH(CH2)2N(CH3)2
OH
253–254
1
2. Chemistry
by H and 13C NMR, and physicochemical property.
The compounds 3, 6, 7, and 18 are the 2-hydroxy tauto-
mers, while the compounds 10–12 exist in 2-oxo forms.
For the biological and physicochemical evaluations the
free bases of 3, 6–8, 10–12, 14, 15, and 18 were converted
into their hydrochloride or dihydrochloride salts by rou-
tine methods.
The synthetic pathways for derivatives 3, 6–8, 10–12, 14,
15, and 18 are shown in Scheme 1. 1-Amino-4-chloro-
anthraquinone (1), the substrate for the majority of syn-
theses, was obtained from 1,4-dichloroanthraquinone
according to Wormser’s method.17 The cyclization of 1
or commercially available 1-aminoanthraquinone (16)
to its anthrapyridone derivatives (2 or 17, respectively)
was performed in the reaction with diethyl malonate
and sodium acetate at an elevated temperature. The sub-
stitution of the 6-chloro residue in compound 2 by an
appropriate amine led to derivative 3 with an 1-ester
group and aminoalkylamino chain at position 6. Com-
pounds 6–8, with basic 1-amido chain and various basic
6-aminoalkylamino arms, were prepared by alkaline
hydrolysis of 2 to acid 4 followed by the reaction with
1,10-carbonyldiimidazole (DCI). The intermediate imi-
dazolide was then directly reacted with N,N-dimethyle-
thylenediamine to give the required amide 5. The
successive displacement of the 6-chloro group by appro-
priate amines carried out at an elevated temperature led
to the desired 6–8. To obtain derivatives unsubstituted
in the pyridone ring (10–12, 14, and 15), the decarboxyl-
ation of 4 by heating its DMA solution was performed.
The resulting 9 on heating with amines readily gave 10–
12. The preparation of derivatives 14 and 15, possessing
2-chloro group instead of 2-hydroxy function at the
pyridone ring, was performed by the reaction of 9 with
phosphonyl chloride, followed by 6-chloro displacement
of resulting 2,6-dichloroanthraquinone (13) with
amines. All attempts to obtain 2,6-disubstituted
aminoalkylamino anthrapyridones failed or, if it were
successful, only poor yields of the products were
achieved. Compound 18, with only 1-carboxamide was
obtained, started with 16, in the analogous reaction used
for the preparation of the intermediates 2, 4, and 5.
3. Results and discussion
The cytotoxicity of anthrapyridone derivatives 3, 6–8,
10–12, 14, 15, and 18, in comparison with reference
clinical drugs doxorubicin (DX) and mitoxantrone
(MIT) against sensitive and resistant human leukemia
cell lines are presented in Table 2 as IC50 values. The
examined compounds are different in the respect to
their structure concerning various functional groups at-
tached to the pyridone ring as well as different amin-
oalkylamino arms at position 6 of the chromophore
moiety. The highest cytotoxic activity among the new
derivatives exhibited compounds 3, 6, and 14. Deriva-
tives 3 and 6 with N,N-dimethylaminoethylamino chain
at position 6 pointed to the effect of cationic 1-carbox-
amide arm in comparison with the 1-ester function.
Compound 3, possessing an ester group, was about
twice more active than 6. Surprisingly, the introduction
of a basic 1-carboxamide moiety to the anthrapyridone
chromophore does not change the activity of the com-
pounds (compare the potency of 7 and 8 with that of
10–12). Derivative 18, holding only an 1-amido chain,
is devoid of cytotoxicity. But converting the 2-hydroxy
to the 2-chloro functionality restores the cytotoxicity
(potency of the compound pair 10 versus 14). The
cytotoxic activity is also affected by the nature of the
aminoalkylamino side chain at position 6. Compounds
with a N,N-dimethylaminoethylamino residue (6, 10,
14) were more potent than those with piperidino moie-
ty (8, 11, 15). The difference in the potency between the
pair 10 versus 12 shows that the optimal distance be-
tween the two nitrogen atoms seems to be two methyl-
ene units.
The structures of the compounds were determined by 1H
and 13C NMR spectroscopy, and by determination of
the molecular weights. The compounds (Table 1) were
proved to occur in two tautomeric forms, as evidenced

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M. Dzieduszycka et al. / Bioorg. Med. Chem. 14 (2006) 2880–2886
O
OH
O
OH
H5C2
(b)
H5C2
O
N
O
N
O
NH2
(a)
O
O
Cl
O
Cl
R
1
(3)
(2)
(1)
R1 = NH(CH2)2N(CH3)2
CH3
N
CH3
O
N
O
OH
OH
O
OH
H3C
N
H
H3C
N
H
N
N
HO
N
(b)
(c)
(d)
(2)
Cl
O
O
O
Cl
R1
(5)
(4)
(6) R1 = NH(CH2)2N(CH3)2
(7) R1 = NH(CH2)2N(C2H5)2
(8) R1 = NH(CH2)2N-c-(CH2)5
OH
OH
N
N
(e)
(b)
(4)
O
Cl
O
R1
(9)
(10) R1 = NH(CH2)2N(CH3)2
(11) R1 = NH(CH2)2N-c-(CH2)5
(12) R1 = NH(CH2)3N(CH3)2
Cl
Cl
N
OH
N
N
(f)
(b)
O
Cl
O
O
Cl
R1
(9)
(13)
(14) R1 = NH(CH2)2N(CH3)2
(15) R1 = NH(CH2)2N-c-(CH2)5
CH3
O
OH
O
OH
N
H5C2
O
NH2
O
N
H3C
N
H
N
(a)
1. (c)
2. (d)
O
O
O
(16)
(17)
(18)
Scheme 1. Synthetic route for compounds 3, 6–8, 10–12, 14, 15, and 18. Reagents and conditions: (a) CH2(COOEt)2, AcONa, 155–160 °C; (b) amine,
D; (c) 20% KOH, EtOH, reflux; (d) CDI, DMA, 80 °C/80 min, amine, rt/18 h; (e) DMA, 165–170 °C/2h; (f) POCl3, PCl5, 100 °C/80 min.
The synthesized anthrapyridones exhibited activity to-
ward the cell lines with induced multidrug cross-resis-
tance of two main types (P-gp and MRP). The
resistance indexes (RI) of the examined compounds were
low (0.65–6.6). Only compound 3 with a 1-ester group
has shown much lower ability to overcome the multi-
drug resistance (RI values are 20 and 50 against
MDR1 and MRP1 cell lines, respectively). No marked
influence of the other substituents on this property could
be observed.
activity against MRP cell line should be stressed, be-
cause such a property is rather rare among cytostatics.
The presence of an ester group at position 1 of pyridone
ring is beneficial for cytotoxic activity but simultaneous-
ly causes the loss of activity in respect to resistant cell
lines. However, the introduction at this position of
aminoalkylamido arm together with an appropriate 6-
aminoalkylamino chain resulted in the appearance of
activity against sensitive as well as resistant cell lines.
On the other hand, lack of a 6-amino side chain affected
a drastic decrease of the cytotoxicity but the ability to
overcome MDR is retained.
In conclusion, anthrapyridone derivatives are the group
of anthraquinone analog antitumor agents with moder-
ate cytotoxicity. These compounds, however, are of
interest because of their ability to overcome multidrug
resistance of tumor cell lines. In particular, the cytotoxic
The obtained results corroborate our hypothesis on the
essential role of heterocyclic ring fused to anthracenedi-

M. Dzieduszycka et al. / Bioorg. Med. Chem. 14 (2006) 2880–2886
2883
Table 2. In vitro cytotoxic activity of examined compounds toward sensitive and resistant human cell lines in comparison with reference doxorubicin
(DX), and mitoxantrone (MIT)
b
Compound
Cell linea/IC50 (nM) SEMc
RId
HL-60
146
HL-60/VINC
HL-60/DX
RId
50.99
3
6
9
2798 395
1012 82
19.21
3.25
1.25
1.00
1.34
1.00
2.26
1.27
0.65
0.95
23.87
33.60
7424 892
667 275
1193 203
1296 201
2092 129
1784 140
311 23
1141 102
1257 86
1159 74
1543 120
1463 295
327 61
2.14
1.05
7
1428 172
1258 121
1551 124
1541 128
3301 262
415 35
8
1.03
1.81
10
11
12
14
15
18
DX
MIT
1.16
2.40
3515
8
2178 363
2113 84
6.66
1.61
1312 126
10,759 79
18.6 0.7
1.0 0.1
853 23
10,235 510
443.9 28.1
33.6 1.8
13,987 31
4163 310
925.1 20.2
1.30
223.82
925.10
a HL-60—human promyelocytic leukemia, and vincristine resistant (MDR1 type) subline HL-60/VINC, and doxorubicin resistant (MRP1 type)
subline HL-60/DX.
b IC50—the concentration of compound causing 50% inhibition of cell growth after 72 h continuous exposure.
c SEM—standard error of the mean.
d RI—resistance index; the ratio of IC50 value for resistant cell line to IC50 value for sensitive cell line.
one compounds in the ability to overcome multidrug
resistance of tumor cells.
3H); 7.96 (dd, 1H, J = 6.6, J = 1.5 Hz); 8.3 (dd, 1H,
J = 1.9 Hz); 12.7 (br s, 1H, ex).
MS m/z (relative intensity, %); 355 [(M+1)+, 100].
4. Experimental
4.1. Chemistry
4.1.2. Ethyl-6[[2-(dimethylamino)ethyl]amino]-2-hydroxy-
7-oxo-7H-naphtho[1,2,3-de]quinoline-1-carboxylate (3).
A sample of 2 with N,N-dimethylethylenediamine was
stirred under reflux in a nitrogen atmosphere for 2 h.
The reaction mixture was diluted with CHCl3 and
washed with diluted HCl to removed excess amine.
The organic layer was flash-chromatographed (eluent
CHCl3/MeOH, 10:1 and 5:1) to give 3 as a dark pink
solid (yield 86%). Mp (as hydrochloride) 253–254 °C.
Melting points were determined with a Boeticus
PHMK05 apparatus and are uncorrected. Combustion
analyses are within +0.4% of the theoretical values and
were carried out on a Carlo Erba CHNS-O-EA1108
instrument for C, H, and N. NMR spectra were taken
on a Varian 300 MHz or 500 MHz spectrometer using
tetramethylsilane as an internal standard. The following
NMR abbreviations were used: ex (exchangeable with
D2O), d ex (exchangeable with D2O, but with difficulty).
Mass spectra were recorded on a Quadrupolic Mass
Spectrophotometer Trio-3 (FAB technique). Thin-layer
chromatography (TLC) was carried out on silica gel
(Kieselgel 60 plates, Merck), column chromatography
was performed on silica gel (Kieselgel Merck, ꢀ200
mesh), if do not marked otherwise, and on Sephadex
LH-20 (Pharmacia).
1H NMR (CDCl3) d 1.3 (t, 3H, J = 7.1 Hz); 2.4 (s, 6H);
2.7 (t, 2H, J = 6.3 Hz); 3.35–3.45 (m, 2H); 4.6 (q, 2H,
J = 7.1 Hz); 7.1 (d, 1H, J = 8.2 Hz); 7.6–7.7 (m, 2H);
7.75 (d, 1H, J = 7.3 Hz), 8.2 (d, 1H, J = 8.3 Hz); 8.6 (d,
1H, J = 7.75 Hz); 10.8 (t, 1H, d ex), 12.8 (br s, 1H, ex).
MS m/z (relative intensity, %): 406 [(M+1)+, 100].
Found: C, 68.0; H, 5.71; N, 10.31. Calcd for
C23H23N3O4: C, 68.13; H, 5.72; N, 10.36.
4.1.1. Ethyl 6-chloro-2-hydroxy-7-oxo-7H-naphtho[1,2,3-
de]quinoline-1-carboxylate (2). A mixture of 250 mg
(2 mmol) of 1, 250 mg of sodium acetate, and 2.5 mL
of diethyl malonate was stirred at 155–160 °C for about
3.5 h. To facilitate the stirring, a small amount of diethyl
malonate was added. The course of the reaction was fol-
lowed by TLC in CHCl3/MeOH (20:1). To the reaction
mixture a portions of ethyl ether and hexane were added
and the resulted yellow-green precipitate was filtered
and then washed with hexane and water. Yield 90%.
An analytical sample was dissolved in warm CHCl3
and flash-chromatographed (using CHCl3/MeOH, 20:1
and 10:1 as eluent). Mp 291–294 °C.
4.1.3. 6-Chloro-2-hydroxy-7-oxo-7H-naphtho[1,2,3-de]quino-
line-1-carboxylic acid (4). A sample of 354 mg (1 mmol) of 2
was dissolved in 22 mL of 20% KOH solution and 18.5 mL
of EtOH. The solution was refluxed for 3.5 h, then cooled
and poured into cold 5% HCl solution. The resulted solid
was separated and dried at 90 °C to give 320 mg of 4, as a yel-
low powder (yield 98%). Mp > 320 °C.
MS m/z (relative intensity, %): 326 [(M)+, 80]; 282
[(Mꢀ44)+].
4.1.4. 6-Chloro-1-[(2-dimethylamino)ethyl]-2-hydroxy-7-
oxo-7H-naphtho[1,2,3-de]quinoline-1-carboxamide (5). A
suspension of 650 mg (2 mmol) of acid 4 in 10 mL of
DMA was stirred at 80 °C for 80 min. Next 500 mg
1H NMR (DMSO-d6) d 1.3 (t, 3H, J = 7.1 Hz); 4.4 (q,
2H, J = 7.1 Hz); 7.64 (d, 1H, J = 8.8 Hz); 7.8–7.93 (m,

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M. Dzieduszycka et al. / Bioorg. Med. Chem. 14 (2006) 2880–2886
(3 mmol) of 1,10-carbonyldiimidazole was added and the
stirring was continued at 40–50 °C for 3 h. After then
the reaction mixture was treated with 1 mL of N,N-
dimethylethylenediamine and the solution was left at
room temperature for 18 h. The course of the reaction
was monitored by TLC in CHCl3/MeOH (10:1). The
solution was diluted with CHCl3, washed with 10%
Na2CO3 solution, water, and dried over Na2SO4. The
solvents were removed and the residue was flash-chro-
matographed (eluent CHCl3/MeOH, 20:1), to afford 5
(79%) as a yellow solid, fluorescing at 366 nm. Mp
218–220 °C.
J = 7.3 Hz); 2.9 (t, 2H, J = 7.3 Hz); 3.05 (m, 2H); 3.5
(t, 2H, J = 6.9 Hz); 6.85 (t, 1H, J = 6.0 Hz, ex); 6.9 (d,
1H, J = 8.8 Hz); 7.52 (t, 1H, J = 7.3 Hz); 7.6 (d, 1H,
J = 8.8 Hz); 7.7 (t, 1H, J = 7.42 Hz); 8.3 (d, 1H,
J = 7.9 Hz); 8.5 (d, 1H, J = 77.81 Hz); 10.45 (t, 1H,
J = 4.88 Hz, d ex); 12.8 (br s, 1H, ex).
MS m/z (relative intensity, %): 476 [(M+1)+ 100].
Found: C, 67.9; H, 6.97; N, 14.70. Calcd for
C27H33N5O3: C, 68.19; H, 6.99; N, 14.73.
4.1.7. 1-[(2-Dimethylamino)ethyl]-6-[[(2-piperidin-1-yl)eth-
yl]amino]-2-hydroxy-7-oxo-7H-naphtho[1,2,3-de]quinoline-
1-carboxamide (8). Compound 8 was obtained in the
reaction of 5 with 1-(2-aminoethyl)piperidine by a proce-
dure similar to that described for 6, as an orange-red sol-
id. Yield 40%. Mp (as dihydrochloride) 115–117 °C.
1H NMR (500 MHz, CDCl3) d 2.28 (s, 6H); 2.4 (t, 2H,
J = 5.3 Hz); 3.0 (q, 2H, J = 5.2 Hz); 6.9 (t, 1H,
J = 4.9 Hz, ex); 7.48–7.56 (m, 3H); 7.7 (t, 1H,
J = 7.8 Hz); 7.83 (d, 1H, J = 8.2 Hz); 7.85 (d, 1H,
J = 7.8 Hz), 12.0 (br s, 1H, ex).
MS m/z (relative intensity, %): 396 [(M)]+, 100].
1H NMR (CDCl3) d 1.5 (m, 2H); 1.7 (m, 4H); 2.25 (s, 6H);
2.45 (t, 2H, J = 5.85 Hz); 2.6 (m, 4H); 2.8 (t, 2H,
J = 6.3 Hz); 3.1 (d, 2H, J = 5.4 Hz); 3.6 (m, 2H); 6.75 (t,
1H, J = 4.8 Hz, ex); 6.95 (d, 1H, J = 8.8 Hz); 7.52–7.58
(m, 2H); 7.7 (t, 1H, J = 7.3 Hz); 8.3 (d, 1H, J = 8.3 Hz),
8.5(d, 1H, J = 7.8 Hz);10.4(m, 1H, ex);11.8(brs, 1H, ex).
4.1.5. 1-[2-(Dimethylamino)ethyl]-6-[[(2-dimethylamino)ethyl]-
amino]-2-hydroxy-7-oxo-7H-naphtho[1,2,3-de]quinoline-
1-carboxamide (6). A sample of 5 with N,N-dimethyl-
ethylenediamine and N,N,N0,N0-tetramethylethylenedi-
amine was stirred at 130 °C in a nitrogen atmosphere
for 6 h. The course of the reaction was monitored by
TLC in CHCl3/MeOH (5:1). The reaction mixture was
diluted with CHCl3 and carefully washed with diluted
HCl to remove excess of amines. The organic layer was
dried over Na2SO4, evaporated and the residue was
flash-chromatographed (eluent CHCl3/MeOH, 10:1 and
5:1). The desired chromatography fractions were collect-
ed and 6 was converted into its dihydrochloride salt
(orange-red powder; yield 20%). The solid was purified
by column chromatography (Sephadex LH-20) eluting
with MeOH. Mp (as dihydrochloride) 296 °C (dec).
MS m/z (relative intensity, %): 488 [(M+1)+, 100].
Found: C, 68.99; H, 6.81; N, 14.32. Calcd for
C28H33N5O3: C, 68.97; H, 6.82; N, 14.36.
4.1.8. 6-Chloro-2-hydroxy-7-oxo-7H-naphtho[1,2,3-de]quino-
line (9). A sample of 200 mg of 4 in 7 mL DMA was stirred
at 165–170 °C for 2h. The progress of the reaction was fol-
lowed by TLC in CHCl3/MeOH (10:1). The reaction mix-
ture was diluted with CHCl3 and washed with 10%
Na2CO3 solution and H2O. The organic layer was dried
over Na2SO4, the solvents were evaporated, and the residue
was flash-chromatographed (eluent CHCl3, then CHCl3/
MeOH, 20:1). Compound 9 was obtained as a yellow-
brown solid (yield 40%), with fluorescence at 366 nm. Mp
164–166 °C (dec).
1H NMR (CDCl3) d 2.25 (s, 6H); 2.4 (s, 6H); 2.45 (t, 2H,
J = 5.8 Hz); 2.7 (t, 2H, J = 5.8 Hz); 2.9 (q, 2H,
J = 5.2 Hz); 3.4 (m, 2H); 6.85 (t, 1H, J = 5.8 Hz, ex);
6.95 (d, 1H, J = 8.5 Hz); 7.5 (t, 1H, J = 7.2 Hz); 7.6 (d,
1H, J = 8.6 Hz); 7.75 (t, 1H, J = 7.4 Hz); 8.32 (d, 1H
J = 7.9 Hz); 8.5 (d, 1H, J = 7.9); 10.45 (t, 1H,
J = 4.9 Hz, ex); 12.8 (br s, 1H, ex).
1H NMR (DMSO-d6) d 7.6 (d, 1H, J = 8.8 Hz); 7.75 (s,
1H); 7.78–7.85 (d, 1H, J = 8.8 Hz overlapping with m,
2H); 8.22 (d, 1H, J = 6.8 Hz); 8.5 (d, 1H, J = 8.3 Hz);
12.8 (br s, 1H, ex).
MS m/z (relative intensity, %): 447 [(M)+, 100].
Found: C, 67.00; H, 6.51; N, 15.60. Calcd for
C25H29N5O3: C, 67.10; H, 6.53; N, 15.65.
MS m/z (relative intensity, %): 282 [(M)+, 100].
4.1.9. 6-[[(2-Dimethyloamino)ethyl]amino]-2-hydroxy-7-
oxo-7H-naphtho[1,2,3-de]quinoline (10). A sample of 9
with N,N-dimethylethylenediamine was stirred at
100 °C in a nitrogen atmosphere for 80 min. The course
of the reaction was monitored by TLC in CHCl3/MeOH
(5:1). The reaction mixture was worked up by the proce-
dure described for the preparation of 6. Further purifi-
cation by flash-chromatography (eluent CHCl3/MeOH,
5:1 and 2:1) gave 10 as a pink solid. Yield 25%. Mp
(as hydrochloride) 230–232 °C (subl).
4.1.6. 1-[2-(Dimethylamino)ethyl]-6[[(2-diethylamino)eth-
yl]amino]-2-hydroxy-7-oxo-7H-naphtho[1,2,3-de]quinoline-
1-carboxamide (7). The reaction of 5 with N,N-diethyle-
thylenediamine was carried out as described for the
preparation of 6; time of the reaction being 6 h. The
reaction mixture was worked up and purified by flash-
chromatography (eluent CHCl3/MeOH, 10:1 and 5:1)
to give 7, as a red solid. Yield 35%. Mp (as dihydrochlo-
ride) 266–268 °C.
1H NMR (CDCl3) d 1.12 (t, 6H, J = 7.3 Hz); 2.52 (s,
3H); 2.54 (s, 3H); 2.54–2.57 (m, 2H); 2.7 (q, 4H,
1H NMR (CDCl3) d 2.4 (s, 6H); 2.8 (m, 2H); 3.5 (m,
2H); 7.7 (m, 5H); 8.2 (m, 1H); 8.5 (m, 1H); 10.6 (br s,

M. Dzieduszycka et al. / Bioorg. Med. Chem. 14 (2006) 2880–2886
2885
1H, ex); 10.75 (br s, 1H, ex); (CDCl3 + TFA) d 3.2 (s,
6H); 3.7 (m, 2H); 4.15 (m, 2H); 7.6 (d, 1H,
J = 9.8 Hz); 7.8–7.85 (m, 1H); 7.9–8.0 (m, 2H); 8.0 (d,
1H, J = 9.3 Hz); 8.1 (s, 1H); 8.4 (m, 1H); 9.4 (br s, 1H).
4.1.13.
2-Chloro-6-[[(2-dimethylamino)ethyl]amino]-7-
oxo-7H-naphtho[1,2,3-de]quinoline (14). A sample of 13
with N,N-dimethylethylenediamine was stirred at
100 °C in a nitrogen atmosphere for 20 min. The course
of the reaction was monitored by TLC in CHCl3/MeOH
(5:1). The reaction mixture was worked up as described
for 6. The flash-chromatography (eluent CHCl3/MeOH,
20:1 then 5:1) gave 14 as a yellow solid strongly fluoresc-
ing at 366 nm. Yield 60%. Mp (as hydrochloride) 260–
261 °C.
Found: C, 72.00; H, 5.75; N, 12.59. Calcd for
C20H19N3O2: C, 72.05; H, 5.74; N, 12.60.
4.1.10.
6-[[(2-Piperidin-1-yl)ethyl]amino]-2-hydroxy-7-
oxo-7H-naphtho[1,2,3-de]quinoline (11). The reaction of
9 with 1-(2-aminoethyl)piperidine was carried out at
80–90 °C for 30 min. The reaction mixture was worked
up by the procedure described for the preparation of 6
and purified by flash-chromatography (eluent CHCl3/
MeOH, 10:1 then 5:1) to afford 11 as a red solid. Yield
32%. Mp (as hydrochloride) 225–227 °C (subl).
1H NMR (CDCl3) d 2.4 (s, 6H), 2.75 (t, 2H, J = 5.8 Hz);
3.5 (m, 2H); 7.78–7.82 (m, 3H); 8.1 (d, 1H, J = 9.6 Hz);
8.4 (d, 1H, J = 7.8 Hz); 8.5 (s, 1H); 8.7 (d, 1H,
J = 7.8 Hz); 11.45 (t, 1H, J = 5.9 Hz, d ex).
Found: C, 68.15; H, 5.12; N, 11.8. Calcd for
C20H18N3OCl: C, 68.28; H, 5.16; N, 11.94.
1H NMR (DMSO-d6) d 1.68–1.72 (m, 2H); 1.72–1.84
(m, 4H); 2.9 (t, 2H, J = 8.2 Hz); 3.3 (t, 2H,
J = 6.9 Hz); 3.5 (m, 2H); 4.0 (t, 2H, J = 6.9 Hz); 7.5 (d,
1H, J = 9.3 Hz); 7.65 (s, 1H); 7.7 (d, 1H, J = 9.3 Hz);
7.72–7.78 (m, 1H); 7.82 (q, 1H, J = 7.3 Hz); 8.35 (d,
1H, J = 5.3 Hz); 8.51 (d, 1H, J = 8.3 Hz); 10.4 (br s,
1H, ex); 10.5 (br s, 1H, ex).
4.1.14. 2-Chloro-6-[[(2-piperidin-1-yl)ethyl]amino]-7-oxo-
7H-naphtho[1,2,3-de]quinoline (15). Compound 15 was
prepared in the reaction of 13 with 1-(2-aminoethyl)pi-
peridine by a procedure similar to that described for
14. Compound 15 is a yellow solid with a strong fluores-
cence at 366 nm. Yield 76%. Mp > 320 °C (dec).
MS m/z (relative intensity, %): 374 [(M+1)+, 100].
1H NMR (DMSO-d6) d 1.6–1.9 (m, 6H); 3.0 (m, 2H); 3.5
(t, 4H, J = 11 Hz); 4.2 (q, 2H, J = 6.8 Hz); 7.8–7.9 (m,
3H); 8.2 (d, 1H, J = 9.7 Hz); 8.5 (d, 1H, J = 7.8 Hz);
8.7 (s, 1H); 8.8 (d, 1H, J = 7.8); 11.5 (t, 1H,
J = 5.9 Hz, d ex).
Found: C, 74.00; H, 6.19; N, 11.24. Calcd for
C23H23N3O2: C, 73.97; H, 6.21; N, 11.25.
4.1.11. 6-[[(3-Dimethylamino)propyl]amino]-2-hydroxy-7-
oxo-7H-naphtho[1,2,3-de]quinoline (12). Compound 12
was prepared in the reaction of 9 with 3-(dimethylami-
no)propylamine by the procedure similar to that de-
scribed for 10, as a pink-orange solid. Yield 26%. Mp
(as hydrochloride) 220–222 °C (subl).
Found: C, 70.45; H, 5.58; N, 10.73. Calcd for
C23H22N3OCl: C, 70.49; H, 5.66; N, 10.72.
MS m/z (relative intensity, %): 392 [(M)+, 100].
1H NMR (CDCl3) d 2.04 (q, 2H, J = 6.8 Hz); 2.4 (s, 6H);
3.0 (t, 2H, J = 7.15 Hz); 3.68 (q, 2H, J = 6.3 Hz), 7.5 (d,
1H, J = 9.1 Hz); 7.63 (s, 1H); 7.68 (d, 1H, J = 9.2 Hz);
7.69–7.75 (m, 1H), 7.8 (q, 1H, J = 7.2 Hz); 8.3 (d, 1H,
J = 5.15 Hz), 8.49 (d, 1H, J = 8.2 Hz); 10.6 (br s, 1H,
ex); 10.7 (br s, 1H, ex).
4.1.15. Ethyl 2-hydroxy-7-oxo-7H-naphtho[1,2,3-de]quin-
oline-1-carboxylate (17). Compound 17 was prepared in
the reaction of 1-aminoanthraquinone (16) with sodium
acetate and diethyl malonate by a procedure similar to
that described for 2. Yield 91%. Mp 310 °C (dec).
1H NMR (DMSO-d6) d 1.3 (t, 3H, J = 7.1 Hz); 4.5 (q,
2H, J = 7.1 Hz); 7.72 (d, 1H, J = 7.1 Hz); 7.83 (t, 1H,
J = 7.3 Hz); 7.86 (d, 1H, J = 8.0 Hz); 7.92 (t, 1H,
J = 2.9 Hz); 8.02 (d, 1H, J = 8.3 Hz); 8.12 (d, 1H,
J = 8.0 Hz); 8.4 (d, 1H, J = 6.0 Hz); 12.7 (br s, 1H, ex).
Found: C, 72.30; H, 6.10; N, 12.05. Calcd for
C21H21N3O2: C, 72.37; H, 6.08; N, 12.06.
4.1.12. 2,6-Dichloro-7-oxo-7H-naphtho[1,2,3-de]quinoline
(13). A sample of 140 mg (0.5 mmol) of 9, 2 mL of
POCl3 and a slightly amount of PCl5 was stirred at
100 °C for 80 min. The course of the reaction was fol-
lowed by TLC in CHCl3/MeOH (10:1); the loss of sub-
strate fluorescence indicating the completion of the
reaction. The unreacted POCl3 was removed under re-
duced pressure and to the residue ice was added. The
mixture was diluted with CHCl3 and washed with 2 N
NaOH solution and H2O. The organic layer was dried
over Na2SO4, evaporated, and the residue was flash-
chromatographed (eluent CHCl3/MeOH, 50:1 then
20:1) to afford 13 as a yellow-brown solid. Yield 60%.
Mp 240–241 °C.
13C NMR (DMSO-d6) d 181.34; 166.90; 159.32; 137.57;
134.16; 132.87; 131.75; 131.63; 131.73; 128.39; 128.24;
126.56; 126.32; 122.49; 121.33; 115.62; 62.15; 13.69.
MS m/z (relative intensity, %): 320 [(M+1)+, 100].
4.1.16. 1-[2-(Dimethylamino)ethyl]-2-hydroxy-7-oxo-7H-
naphtho[1,2,3-de]quinoline-1-carboxamide (18). A sample
of 17 was converted into acid by the same manner as de-
scribed for 4. The obtained 2-hydroxy-7-oxo-7H-naph-
tho[1,2,3-de]quinoline-1-carboxylic acid in DMA
solution was then treated with 1,10-carbonyldiimidazole
and the intermediate imidazolide was reacted with N,N-
dimethylethylenediamine. The procedure of the reaction
MS m/z (relative intensity, %): 301 [(M)+, 100].

2886
M. Dzieduszycka et al. / Bioorg. Med. Chem. 14 (2006) 2880–2886
was similar to that described for 5. Yield 80%. Mp (as
hydrochloride) 253–254 °C.
Acknowledgments
The authors acknowledge the financial support of these
studies by the State Committee for Scientific Research
(KBN), Warsaw, Poland (Grant No. 4 P05F 035 19),
and in part by Italian Ministry of Foreign Affairs and
´
the Chemical Faculty Gdansk University of Technology,
Poland.
1H NMR (CDCl3) d 2.3 (s, 6H); 2.45 (t, 2H, J = 6.0 Hz);
3.55 (q, 2H, J = 7.6 Hz); 6.45 (t, 1H, ex); 7.3–7.7 (m,
3H); 7.8 (t, 1H, J = 7.2 Hz); 8.18 (d, 1H, J = 7.5 Hz);
8.34 (dd, 1H, J = 7.1, 1.6 Hz); 8.5 (dd, 1H, J = 6.7,
1.36 Hz); 11.8 (br s, 1H, ex).
MS m/z (relative intensity, %): 362 [(M+1)+, 100].
References and notes
Found: C, 69.74; H, 5.26; N, 11.62. Calcd for
C21H19N3O3: C, 69.79; H, 5.30; N, 11.63.
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