close

Вход

Забыли?

вход по аккаунту

?

New Derivatives of Quinazoline and 1 2-Dihydroquinazoline N3-Oxide with Expected Antitumor Activity.

код для вставкиСкачать
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 239−246
Elżbieta Mikiciuk-Olasika,
Katarzyna BłaszczakŚwia˛tkiewiza,
Elżbieta Żureka,
Urszula Krajewskab,
Marek Różalskib,
Rafał Kruszyńskic,
Tadeusz J. Bartczakc
a
Department of
Pharmaceutical Chemistry
and Drug Analysis,
Medical University, Łódź,
Poland
b
Department of Biochemistry,
Medical University,
Łódź, Poland
c
X-Ray Crystallography and
Crystal Chemistry Group,
Institute of General and
Ecological Chemistry,
Łódź University of
Technology, Łódź, Poland
New Derivatives with Antitumor Activity 239
New Derivatives of Quinazoline and 1,2Dihydroquinazoline N 3-Oxide with Expected
Antitumor Activity
Some derivatives of quinazoline and 1,2-dihydroquinazoline N 3-oxide were synthesised and their cytotoxic activities against human leukaemia HL-60 cells under hypoxic and aerobic conditions were tested and compared to tirapazamine
as the reference compound. Compound 8 showed 5-fold higher toxicity under
hypoxic conditions than in normal oxygen atmosphere. The structure of quinazoline derivatives was established by X-ray crystal structure analysis.
Keywords: Quinazoline; 1,2-Dihydroquinazoline N 3-oxides; Hypoxia; Cytotoxic
activity
Received: November 29, 2001; Accepted: January 9, 2004 [FP656]
DOI 10.1002/ardp.200100656
Introduction
Cancer is one of the most mysterious diseases and
challenges of medicine at the beginning of the twenty
first century. The dynamic development of science and
the research on cancer pathophysiology are associated with studying of novel targets for pharmacotherapy. The typical feature of solid tumors is hypoxia and
this feature is used not only in oncological diagnostics
[1] but also in anticancer therapy [2⫺4].
The hypothesis of bioreductive activation of drugs in
hypoxic tissues was confirmed by research data,
which allowed selecting several compounds with cytotoxic activity and selective affinity to pathologic tumour
cells. The biological activity of these pharmacological
groups is attributed to oxidizing properties. These are
the compounds: quinone antibiotics (e.g. mitomycine
C), nitroimidazole derivatives (e.g. metronidazole, misonidazole), 1,2,4-benzotriazine di-N-oxides (e.g. tirapazamine) and quinoxaline 1,4-di-N-oxides. Nitroimidazole derivatives, beside a selective cytotoxic activity, play the role of substances that sensitize tumour
tissue to radiotherapy [4].
Correspondence: Elżbieta Mikiciuk-Olasik, Department of
Pharmaceutical Chemistry, Medical University of Łódź,
Muszyńskiego 1, 90-151 Łódź, Poland. Phone: +48 42 67792-50, Fax: +48 42 677-92-50; e-mail: eolasik@farm.
pharm.am.lodz.pl
Pharmacological activity of tirapazamine and the derivatives of quinoxaline 1,4-di-N-oxide was the beginning of the search for novel chemical compounds
among the derivatives of 1,2-dihydroquinazoline N 3oxide.
The reaction of E-amino-oximes with aldehydes has
been previously described [5, 6]. We have designed
the synthesis of new derivatives of quinazoline and
1,2-dihydroquinazoline N 3-oxide as the compounds
with expected cytotoxic activity and selective affinity to
hypoxic tumour tissues.
The subject of this study was the synthesis of novel
quinazoline derivatives (8⫺13). Compounds 8⫺10
were synthesized by direct condensation of 2-amino5-chlorobenzophenone E-oxime (1) with appropriate
aldehydes (3, 4) or ketone (5). Because of not successful attempts of synthesising Schiff’s base from
2-amino-5-chlorobenzophenone E-oxime (1) with
butanedione monooxime (5) in room temperature, we
decided to obtain compound 10 only by direct way, this
means, by heating 2-amino-5-chlorobenzophenone Eoxime (1) with butanedione monooxime (5). However,
compounds 8 and 9 were also synthesized by an indirect way, which involved cyclocondensation of previously prepared derivatives of 2-(N-methylideneamine)benzophenone oxime (6, 7). The latter compounds
(6, 7) are a new class of ligands for technetium-99m
complexes. Complex 99mTc-7, due to its lipophilic
properties, is a potential radiopharmaceutical for brain
imaging [7]. As it appeared, in Scheme 1, compounds
 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
240 Mikiciuk-Olasik et al.
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 239−246
Scheme 1. Synthesis of compounds 8⫺13. Reagents: i ⫺ CH3COOH, room temp.; ii ⫺ EtOH, CH3COOH, ∆;
iii ⫺ CH3COOH.
6 and 7 are intermediates in the presented synthesis
of 1,2-dihydroquinazoline N 3-oxide. Analogues of 1,2dihydroquinazoline N 3-oxide (11⫺13) were synthesized by cyclocondensation of 2-amino-5-chlorobenzophenone E-oxime (1) or 2-amino-5-nitrobenzophenone E-oxime (2) with appropriate aldehydes
(3, 4) or a ketone (5) in different conditions.
Selected compounds were tested as potential cytotoxic species. The cytotoxicity of quinazoline (11, 12)
and 1,2-dihydoquinazoline N 3-oxide derivatives
(8⫺10) against human promyelocytic leukaemia
HL-60 and lymphoblastic leukaemia NALM-6 cells was
tested after chronic exposure (3 days) using a haemocytometer for cell counts and the trypan-blue exclusion
method to determine cell viability. Additionally, the effect of 1,2-dihydroquinazoline N 3-oxide derivatives
(8, 9 and 10) on the survival of HL-60 leukaemia
cells under normoxic and hypoxic conditions was
measured.
Tested 1,2-dihydoquinazoline N 3-oxides derivatives
showed higher cytotoxic activity than their quinazoline
analogues. Compounds 8 and 9 exhibited relatively
powerful cytotoxic effect, but compound 10 showed
only moderate cytotoxic activity during three days
chronic exposure. Moreover, compound 8 showed
higher toxicity under hypoxic conditions than in normal
oxygen atmosphere. Hypoxic/aerobic toxicity coefficient was ca. 5 (Table 1).
 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 239−246
New Derivatives with Antitumor Activity 241
Table 1. IC50-Values for HL-60 cell line exposed to N3oxides of quinazoline and tirapazamine under oxic and
hypoxic conditions.
Compound
IC50†
Oxic (O)
‡
12
11
9
8
10
Tirapazamine
0.8
5.2
0.9
10.2
IC50 Values in µM (mean ± SD, n ⱖ 6). Cells were
exposed for 4 h to the tested compounds, then the
culture medium was exchanged and a viability assay
was done 48 h after the beginning of experiment.
Statistically significant differences.
IC50†
Compound
Differential
toxicity
Hypoxic (H)
O/H
9
48.8±13.9 58.4±7.4
8
478.8±91.9 92.8±8.7‡
10
513.2±16.9 551.1±39.0
Tirapazamine 87.8±3.0
8.6±1.2‡
†
Table 2. Cytotoxicity of tested compounds for human
leukaemia HL-60 and NALM-6 cells in chronic exposure.
†
HL-60
NALM-6
465.6±47.0
176.5±55.3
57.0±8.1
37.0±5.7
445.2±54.2
3.4±1.3
192.7±32.9
95.6±2.8
53.2±3.2
57.9±5.0
57.8±7.9
7.0±1.9
IC50 Values in µM (mean ± SD, n ⱖ3).
boiling acetic acid as a solvent. All syntheses were
monitored by thin layer chromatography.
Further research of these compounds should determine their possible usefulness in treatment of cancer
diseases.
The structure of the lead compound (12) was established by X-ray crystal structure analysis. It confirms
the presence of quinazoline ring with naphthalene substituent at the 2-position. The quinazoline and naphthalene ring systems are almost coplanar. This conformation is stabilised by two weak hydrogen bonds.
Results and discussion
Based on our study we determined two methods for
synthesising of derivatives of 4-phenyl-1,2-dihydroquinazoline N 3-oxide. The first one involved direct condensation of 2-amino-5-chlorobenzophenone E-oxime
(1) with: piperonal (3) or 2-naphthaldehyde (4) or butanedione monooxime (5). Another method involved
cyclisation of previously prepared Schiff’s bases
(6, 7)⫺potential ligands for Technetium-99m [7]. We
have elaborated conditions for the synthesis of derivatives of 4-phenyl-1,2-dihydroquinazoline N 3-oxide by
direct (8⫺10), and indirect (8, 9), cyclocondensation.
Furthermore, we established conditions for preparing
of the derivatives of 5-chloro-2-(N-arylmethylideneamine)benzophenone oxime (6 and 7) ⫺ intermediates for
synthesising 2-aryl-6-chloro-4-phenyl-1,2-dihydroquinazoline N 3-oxide (8⫺10). We also performed cyclocondensation of 2-amino-5-chlorobenzophenone Eoxime (1) or 2-amino-5-nitrobenzophenone (2) with aldehydes (3, 4) to obtain novel analogues of quinazoline (11⫺13) ⫺ another new class of planned compounds. These compounds (11⫺13) were prepared in
Cytotoxicity in normoxia
Newly synthesised compounds (8⫺12) and tirapazamine as reference compound have been evaluated in
chronic cytotoxicity experiments.
Their biological activity was determined in vitro on human leukaemia HL-60 and NALM-6 cell lines in optimal atmosphere with 95 % air/5 % CO2. IC50 values of
compounds 8⫺12 were over the range 37-465 µM
(Table 2). Quinazoline derivatives (compounds 11 and
12) presented rather low cytotoxic activity against both
cell lines. More efficient in cell killing were derivatives
of 1,2-dihydroquinazoline N 3-oxides (compounds 8, 9
and 10). The measured IC50 value for the most potent
1,2-dihydroquinazoline N 3-oxide with piperonyl moiety
(8) was 37±5.7 µM for HL-60 cells. Generally, both
leukaemia cell lines were almost equally sensitive to
derivatives of 1,2-dihydroquinazoline N 3-oxide with
the exception of 6-chloro-4-phenyl-2-methyl-2-[(1hydroxyimino)ethyl]-1,2-dihydroquinazoline N 3-oxide
(10), which was ca. 7-fold less cytotoxic against
HL-60 than for NALM-6 cells.
Cytotoxicity in hypoxia
Compounds 8⫺10 were initially chosen for biological
assessment in this study and comparison with 1,2,4benzotriazin-3-amine 1,4-dioxide (tirapazamine). After
a 4-h exposure, a selective cytotoxicity (higher in hypoxia than at normal oxygen conditions) of 6-chloro-4phenyl-2-piperonyl-1,2-dihydroquinazoline N 3-oxide
(8) against HL-60 cells was observed (Table 1). The
 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
242 Mikiciuk-Olasik et al.
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 239−246
hypoxic/aerobic toxicity coefficient was 5.2. This parameter locates the tested compound between mitomycine C (toxicity coefficient from 1-5) and 2-nitroimidazole (misonidazole) with toxicity coefficient from 5⫺15
[2, 4].
X-Ray crystallography results and discussion
The perspective view of the lead compound (12), together with the atom-numbering scheme is shown in
Figure 1. All interatomic distances can be considered
as normal. All three-ring systems are planar within the
experimental error. The maximum sticking out atoms
C4, C15 and C9 deviate from the weighted root mean
squares planes of quinazoline (Cremer and Pople total
puckering amplitude [8] QT = 0.092 Å), naphtalene (total puckering amplitude QT = 0.072 Å), and benzene
for 0.0485 Å (17), 0.0337 Å (17) and 0.0154 Å (15),
respectively. The adjacent atoms to the quinazoline
ring system C(15), C(9) and Cl(1) deviate from its
weighted root mean squares plane by 0.088(3),
⫺0.113(3) and 0.035(2) Å respectively. The quinazoline and benzene rings are inclined at 45.50(6)°. The
quinazoline and naphthalene ring systems are almost
coplanar, and they determine an angle of 3.26(7)°.
This conformation is stabilised by two weak hydrogen
bonds [9⫺11] linking C16⫺H16···N(2) (D···A distance
of 2.824(3) Å and D⫺H···A angle of 100.5°) and
C(24)⫺H(24)···N(1) atoms (D···A distance of 2.788(3)
Å and D-H···A angle of 101.2°). For the weighted root
mean squares plane calculated for all atoms of quinazoline and naphthalene the maximum deviation of
0.086(2) occurs for C(22) atom. The molecules of 6chloro-2-(2-naphthyl)-4-phenylquinazoline (12) are
held together via two types of interactions: π-π stacking and C⫺H···π weak hydrogen bonds. Stacking interactions can be divided into two types: (i) between repeating quinazoline rings (Table 3, Figure 2) and (ii)
between repeating quinazoline and naphthalene rings
(Table 3, Figure 2). The benzene ring does not take
part in the stacking scheme. The fragment of crystal
packing showing stacking intermolecular interactions
is depicted in Figure 2. Intermolecular weak hydrogen
bonds link C(13)⫺H(13)···Cg(5# x+1/2, -y+1/2, z-1/2)
(D···A distance 3.585 Å and D⫺H···A angle of 129.5°)
and C(20)-H(20)···Cg(3# -x+1/2, y+1/2, -z+1/2) (D···A
distance equal to 3.832 Å and D-H···A angle equal to
121.2°) atoms and π-systems; rings are numbered
according to Figure 3. Thus, the three-dimensional infinite net structure is created. There are no unusually
short intermolecular contacts except the mentioned
hydrogen bonds and stacking interactions.
Figure 1. Molecular structure of compound 12.
Displacement ellipsoids are drawn at the 50 %probability level. Intramolecular hydrogen bonds are
indicated by dashed lines.
Figure 2. Molecular packing of 12 showing the arrangement of stacking rings.
Acknowledgments
Synthetic research is supported by the Medical University of Łódź, Poland (grant No 507 13 028). The biochemical part of this work is supported by the Medical
University of Łódź, Poland (grant No 502-13-843
(192)). The crystallographic part of this work was financed by the funds allocated by the State Committee
for Scientific Research to the Institute of General and
Ecological Chemistry, Poland, Łódź University of Technology.
 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 239−246
New Derivatives with Antitumor Activity 243
son Infinity Series FT-IR spectrophotometer (USA). 1H and
13
C spectra were recorded on a 300 MHz Varian Mercury
spectrometer (Germany) in DMSO or CDCl3 as solvent and
tetramethylsilane (TMS) as internal reference. MS spectra
(FAB method, M+1, matrix⫺glycerine) were recorded on a
Finnigan Mat 95 spectrometer (Brema, Germany). Carbon,
hydrogen and nitrogen elemental analyses were performed
using the Perkin Elmer 2400 series II CHNS/O (Madison,
USA), and agreed with proposed structures within ±0.3 % of
theoretical values.
Chromatographic purification was performed on HPTLC and
silica gel plates (Merck F254, Darmstadt, Germany) with indicated eluents. Chemicals and solvents were obtained from
commercial sources. Benzophenone oxime derivatives were
obtained according to standard methods [12, 13].
General procedure for preparation of compounds 6 and 7
A mixture of equimolar portions of the appropriate oxime (1)
(10 mmol) and of the appropriate aldehyde (3⫺4), (10 mmol)
was dissolved in anhydrous acetic acid (50 mL) and stirred
at room temperature. After 24 h, the precipitated crude yellow
product was filtered off. Recrystallization from isopropanol
gave the compounds 6 and 7 ⫺ Schiff’s bases with chromatographic purity (chloroform/methanol⫺6.25 % v/v).
Figure 3. Chemical structure of compound 12 with
ring numbering.
Experimental
Procedures
Melting points were measured on the Electrothermal apparatus (Electrothermal, England) and are presented uncorrected. IR spectra (KBr discs) were registered using the Matt-
General procedure for preparation of compounds 8⫺10
Starting from 2-aminobenzophenone oxime (1): the mixture
of equimolar portions of the oxime (1) (10 mmol) and of the
appropriate aldehyde (3⫺4), (10 mmol) or ketone (5), (10
mmol) in anhydrous ethanol (50 mL) containing acetic acid
(5 mL as a catalytic agent) was heated under reflux. After
2 h the reaction mixture was neutralised using 10 % KOH
(ethanol solution) to pH = 7 and concentrated to the half of
its initial volume. A crude precipitate was filtered off. The dry
precipitate was collected and recrystallized from acetonitrile.
Table 3. Stacking interactions parameters. Rings are numbered according to Figure 3.
Interacting
rings
Distance between Dihedral angle Angle between linking ring Perpendicular distance of
ring centroids between planes
centroids vector and
the first ring plane on the
normal to the first ring plane
second ring plane
(Å)
(deg)
(deg)
(Å)
Cg(1)⫺Cg(2)#1
Cg(2)⫺Cg(1)#1
4.2024
4.2024
2.64
2.64
31.40
31.69
3.576
3.587
Cg(2)⫺Cg(2)#1
Cg(1)⫺Cg(1)#2
3.6182
4.4342
0.00
0.02
8.09
35.15
3.582
3.626
Cg(1)⫺Cg(4)#2
Cg(4)⫺Cg(1)#2
Cg(1)⫺Cg(5)#2
Cg(5)⫺Cg(1)#2
3.9522
3.9522
5.2964
5.2964
1.75
1.75
3.70
3.70
25.04
25.83
47.29
49.23
3.557
3.581
3.459
3.593
Cg(2)⫺Cg(4)#2
Cg(4)⫺Cg(2)#2
Cg(2)⫺Cg(5)#2
Cg(5)⫺Cg(2)#2
3.7252
3.7252
4.5413
4.5413
3.97
3.97
4.92
4.92
18.30
14.97
40.61
37.65
3.599
3.537
3.596
3.448
Used symmetry transformations: #1 -x,-y,-z; #2 -x,-y+1,-z.
 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
244 Mikiciuk-Olasik et al.
Chromatographic purity of the obtained compounds was confirmed using a mixture: ethyl acetate/acetonitrile 25 % (v/v) as
eluent for compounds 8⫺9 or ethyl acetate/methanol 25 %
(v/v) for compound 10.
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 239−246
N 7.39; found: C 66.59; H 4.09; N 7.60; Rf (ethyl acetate/
acetonitrile 25 % (v/v)) = 0.47.
6-Chloro-2-(2-naphthyl)-4-phenyl-1,2-dihydroquinazoline N3oxide (9)
General procedure for preparation of compounds 8 and 9
Starting
from
5-chloro-2-(N-methylideneamine)benzophenone oxime (6, 7): A mixture of equimolar portions of the
appropriate oxime (6, 7), (10 mmol) in anhydrous acetic acid
(50 mL) was heated under reflux for 24 h. The solution was
then evaporated to the half of its initial volume and a crude
precipitate was filtered off. The dry solid was recrystallized
from isopropanol. Chromatographic purity of the obtained
compounds was confirmed using a mixture: ethyl acetate/
acetonitrile 25 % (v/v) as eluent (8⫺9).
General procedure for preparation of compounds 11⫺13
From 2-aminobenzophenone oxime (1⫺2): The mixture of
equimolar portions of the appropriate oxime (1, 2) (10 mmol)
and of the appropriate aldehyde (3, 4), (10 mmol) in anhydrous acetic acid (50 mL) was heated under reflux. After 24
h, the reaction mixture was concentrated to the half of its
initial volume and a crude precipitate was filtered off. The dry
precipitate was collected and recrystallized from acetonitrile.
Chromatographic purity of the obtained compounds was confirmed using a mixture: chloroform/ethyl acetate 5 % (v/v) as
eluent.
2-(N-benzo[1,3]dioxol-5-yl-methylideneamine)-5-chlorobenzophenone oxime (6)
Yield 70 %, mp 140 °C; IR (KBr) ν/cm⫺1: 3162 (NOH), 3020
(ArH), 1635 (C=N), 1260 (C-O-Casym), 1034 (C-O-Csym); 1H
NMR (DMSO) δ 11.30 (s, 1H, NOH), 8.30 (s, 1H, N=CH),
7.55⫺7.49 (dd, 4H, 4JHH = 2.4 Hz, 4JHH = 2.4 Hz CHarom),
7.38⫺7.19 (m, 5H, CHarom), 7.05 (d, 4JHH = 1.4 Hz, 1H,
CHarom), 6.97 (d, 3JHH = 8.0 Hz, 1H, CHarom), 6.0 (s, 2H, CH2);
MS m/z: 379, 381; calculated for C21H15N2O3Cl: C 65.58; H
3.99; N 7.39; found: C 66.38; H 3.94; N 7.41; Rf (chloroform/
methanol ⫺ 6.25 % v/v) = 0.45.
5-Chloro-2-[N-(2-naphhtylmethylideneamine)]benzophenone
oxime (7)
Yield 72 %; mp 168 °C; IR (KBr) ν/cm-1: 3221 (NOH), 3057
(ArH), 1614 (C=N); 1H NMR (DMSO) δ 11.36 (s, 1H, NOH),
8.63 (s, 1H, N=CH), 8.21 (s, 1H, CHarom), 8.00⫺7.87 (mm,
4H, CHarom), 7.61⫺7.59 (d, 3JHH = 8.5 Hz, 1H, CHarom),
7.58⫺7.54 (m, 2H, CHarom), 7.41⫺7.31 (m, 5H, CHarom); MS
m/z: 385, 387; calculated for C24H17N2Cl: C 74.9; H 4.45; N
7.28; found: C 74.36; H 4.96; N 6.84. Rf (chloroform/methanol
⫺ 6.25 % v/v) = 0.53.
2-Benzo[1,3]dioxol-5-yl-6-chloro-4-phenyl-1,2-dihydroquinazoline N3-oxide (8)
Yield (70 %); mp 204 °C; IR (KBr) ν/cm⫺1: 3213 (NH), 3032
(ArH), 1487 (C=N), 1264 (N-O) 1240 (C-O-Casym), 1037 (CO-Csym); 1H NMR (DMSO) ( 7.95 (d, 3JHH = 2.7 Hz, 1H, NH),
7.56⫺7.41 (m, 5H, CHarom), 7.17 (dd, 4JHH = 2.3 Hz, 1H,
CHarom), 6.99⫺6.95 (m, 3H, CHarom), 6.92 (d, 3JHH = 7.9 Hz,
1H, CHarom), 6.48 (d, 4JHH = 2.3 Hz, 1H, CHarom), 6.16 (d,
4
JHH = 2.7 Hz, 1H, CH), 6.0 (s, 2H, CH2); 13C NMR (DMSO)
δ (ppm): 79.58, 101.29, 106.57, 108.08, 116.22, 118.91,
119.92, 120.37, 122.21, 124.14, 129.10, 129.36, 129.83,
130.10, 131.70, 137.67, 139.34, 147.45, 147.59; MS m/z:
379.1, 381.1; calculated for C21H15N2O3Cl: C 66.58; H 3.99;
Yield (75 %); mp 219 °C; IR (KBr) ν/cm⫺1: 3216 (NH), 3058
(ArH), 1490 (C=N), 1271 (N-O); 1H NMR (CDCl3) δ (ppm):
7.95 (d, 4JHH = 0.59 Hz, 1H, CHarom), 7.85⫺7.74 (m, 4H,
CHarom) 7.50⫺7.45 (m, 7H, CHarom), 7.11 (d, 3JHH = 2.2 Hz,
1H, CHarom), 6.84 (d, 3JHH = 8.5 Hz, 1H, CHarom), 6.76 (d,
4
JHH = 2.2 Hz, 1H, CHarom), 6.35 (d, 4JHH = 3.9 Hz, 1H, CH),
5.22 (d, 4JHH = 3.5 Hz, 1H, NH exchange. with D2O). 13C
NMR (DMSO) δ: 79.99, 116.39, 119.09, 122.35, 124.09,
124.25, 125.59, 126.66, 126.71, 127.58, 128.16, 128.46,
128.62, 129.22, 129.47, 129.93, 130.16, 132.38, 132.95,
135.29, 138.25, 139.48; MS m/z: 385.2; 287.2; Calculated for
C24H17N2OCl: C 74.90; H 4.45; N 7.28; Found: C 74.82; H
4.13; N 7.30; Rf (ethyl acetate/acetonitrile 25 % (v/v)) = 0.49.
6-Chloro-4-phenylo-2-methyl-2-[1-hydroxyimino)ethyl]-1,2dihydroquinazoline N3-oxide (10)
Yield 60 %, mp 160 °C, IR (KBr) ν/cm⫺1: 3215 (NH), 3169
(NOH), 3067 (ArH), 1479 (C=N), 1257 (N씮O); 1H NMR
(DMSO) δ (ppm): 11,16 (s, 1H, NOH), 7.84 (s, 1H, NH exchange. with D2O), 7.49⫺7.51 (m, 3H, CHarom), 7.47 (d,
3
JHH = 6.8 Hz, 1H, CHarom), 7.13 (dd, 4JHH = 2.1 Hz, 1H,
CHarom), 6.85 (d, 3JHH = 8.5 Hz, 1H, CHarom), 1.82 (s, 3H,
CH3), 1.71 (s, 3H, CH3); 13C NMR (DMSO) δ (ppm): 9.99,
21.54, 83.00, 115.67, 118.83, 121.84, 123.67, 128.41,
128.76, 129.10, 129.79, 130.90, 137.96, 139.27, 153.02; MS
m/z: 330.1 332.1; calculated for C17H16N3O2Cl ·H2O: C ⫺
58.71, H ⫺ 5.22, N ⫺ 12.08; found: C ⫺ 58.26, H -5.19, N ⫺
12.81; Rf (ethyl acetate/methanol25 % (v/v)) = 0.54.
2-Benzo[1,3]dioxol-5-yl-6-chloro-4-phenylquinazoline (11)
Yield 50 %; mp 200 °C; IR (KBr) ν/cm⫺1: 3082 (ArH), 1556
(C=N), 1229 (C-O-Casym), 1035 (C-O-Csym); 1H NMR
(CDCl3) δ 8.29 (dd, 4JHH = 1.8 Hz, 1H, CHarom), 8.16 (d,
4
JHH = 1.8 Hz, 1H, CHarom), 8.07 (s, 1H, CHarom), 8.04 (d,
3
JHH = 9.1 Hz, 1H, CHarom), 7.86⫺7.77 (m, 3H, CHarom),
7.63⫺7.59 (m, 3H, CHarom), 6.94 (d, 3JHH = 8.3 Hz, 1H,
CHarom), 6.05 (s, 2H, CH2); 13C NMR (CDCl3) δ (ppm):
101.45, 108.28, 108.77, 121.95, 125.73, 125.76, 128.70,
130.00, 130.17, 130.65, 132.20, 132.25, 134.43, 137.12,
148.14, 150.03, 150.49, 159.92, 167.35; MS m/z: 361.0, 363;
Calculated for: C21H13N2O2Cl: C ⫺ 69.91, H ⫺ 3.63, N ⫺
7.76; Found: C ⫺ 70.17, H ⫺ 3.39, N ⫺ 7.71; Rf (chloroform/
ethyl acetate 5 % (v/v)) = 0.53.
6-Chloro-2-(2-naphthyl)-4-phenylquinazoline (12)
Yield 60 %; mp181 °C; IR (KBr) ν/cm⫺1: 3055(ArH), 1536 (C=
N); 1H NMR (CDCl3) δ 9.22 (d, 4JHH = 1.8 Hz, 1H, CHarom),
8.78 (dd, 4JHH = 1.8 Hz, 1H, CHarom), 8.17 (d, 3JHH = 9.1 Hz
1H, CHarom), 8 Hz, 1H, CHarom), 12⫺7,83 (m, 7H, CHarom),
7.67⫺7.64 (m, 3H, CHarom), 7.56⫺7.52 (m, 2H, CHarom); 13C
NMR (CDCl3) δ (ppm): 122.18, 125.41, 125.77, 126.19,
127.08, 127.69, 128.15, 128.73, 129.09, 129.26, 130.06,
130.2, 130.8, 132.56, 133.33, 134.45, 134.74, 135.08, 137.1,
150.47, 160.35, 167.50; MS m/z: 367.2; 369.2; calculated for
C24H15N2Cl: C ⫺ 78.58, H ⫺ 4.12, N ⫺ 7.64; found: C ⫺
77.99, H ⫺ 3.84, N ⫺ 7.85; Rf (chloroform/ethyl acetate 5 %
(v/v))= 0.66.
 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 239−246
New Derivatives with Antitumor Activity 245
2-Naphthyl-6-nitro-4-phenylquinazoline (13)
X-ray structure analysis
⫺1
Yield 65 %, mp 190 °C, IR (KBr) ν/cm : 3094 (ArH), 1581
(C=N), 1539 (NO2 asym), 1338 (NO2 sym); 1H NMR (CDCl3) δ
9.1 (s, 1H, CHarom), 8.96 (d, 4JHH = 2.6 Hz, 1H, CHarom), 8.70
(dd, 4JHH = 1.6 Hz, 1H, CHarom), 8.55 (dd, 4JHH = 2.6 Hz, 1H,
CHarom), 8.19 (d, 3JHH = 9.1 Hz, 1H, CHarom), 7.97⫺7.81 (m,
5H, CHarom), 7.65⫺7.1 (m, 3H, CHarom), 7.52⫺7.43 (m, 2H,
CHarom); 13C NMR (CDCl3) δ (ppm): 120.42, 124.21, 125.42,
126.42, 126.93, 127.68, 127.76, 128.36, 129.10, 129.43,
130.08, 130.28, 130.91, 130.97, 133.19, 134.37, 135.13,
136.39, 145.32, 154.46, 162.73, 170.39; MS m/z: 378.2; calculated for C24H15N2O2: C ⫺ 76.38, H ⫺ 4.01, N ⫺ 11.13;
found: C ⫺ 76.07, H ⫺ 3.63, N ⫺ 10.95; Rf (chloroform/ethyl
acetate 5 % (v/v)) = 0.58.
A rectangular prism crystal of 6-chloro-2-(2-naphthyl)-4-phenylquinazoline of approximate dimensions 0.097 ⫻ 0.167 ⫻
0.329 mm was mounted on a KM-4-CCD automatic diffractometer equipped with a CCD detector (Oxford Diffraction
Poland, Wroclaw, Poland) and used for data collection. X-ray
intensity data were collected with graphite monochromatic
MoKα radiation (λ = 0.71073 Å) at a temperature of 289.0(2)
K and ω-scan mode. A 24-seconds exposure time was used.
The unit cell parameters were determined from least-squares
refinement of the setting angles of 2215 strongest reflections.
Examination of two reference frames monitored after each 20
frames measured showed 2.17 % loss of the intensity. During
the data, reduction above decay correction coefficient was
taken into account. A yellow, transparent crystal used for data
Assay monitoring cytotoxic effects of compounds 8⫺12 and
tirapazamine on human leukaemia HL-60 and NALM-6 cells
Cell culture
Human acute myeoloblastic leukaemia HL-60 cells and lymphoblastic leukaemia NALM-6 cells were used. The NALM-6
cell line was purchased from the German Collection of Microorganisms and Cell Cultures [14].
Cells were cultured in RPMI 1640 medium supplemented with
10 % heat-inactivated fetal calf serum and 2 mM glutamine,
at 37 °C in a 95 % air/5 %CO2 atmosphere.
Cytotoxicity
Exponentially growing HL-60 and NALM-6 cells were seeded
at 0.4 ⫻ 106 per each well of a 24-well plate (Nunc, Roskilde,
Denmark) and cells were then exposed to the compounds
indicated in the experimental section. Stock solutions of the
test compounds were prepared freshly in DMSO and were
used for serial dilutions in complete culture medium. Final
concentration of DMSO in medium was 0.2 %. In the case of
chronic exposure, cells were incubated in a 95 % air/5 % CO2
atmosphere for three days. Cytotoxic activity of the new derivatives of quinazoline and 1,2-dihydroquinazoline N 3-oxides
was compared to activity of tirapazamine. The number of viable cells was counted in a Bürker haemocytometer using a
trypan-blue exclusion assay, and dose-response curves were
determined. Values of IC50 (the concentration of tested compounds required to reduce leukaemia cells survival fraction
to 50 % of control) was used as a measure of cellular sensitivity to a given treatment.
Three derivatives of 1,2-dihydroquinazoline N 3-oxide and tirapazamine as reference compound were also evaluated
using trypan-blue exclusion assay to determine IC50 values
under oxic and hypoxic conditions.
For experiments with low oxygen atmosphere (hypoxia) 6well plates containing 0.4 ⫻ 106 HL-60 cells/well were posed
into a glass leak-proof chamber connected to inlet lines for air
or nitrogen. Hypoxia was achieved by gassing with nitrogen
containing 50 ppm of oxygen (0.04 mmHg). After 4 h exposure in air or nitrogen, cells were washed free of drugs with
medium and allowed to grow for two days in fresh culture
medium (in 95 % air/5 % CO2). Results were averaged for six
independent experiments.
Statistical analysis of the data
The results are expressed as means ± SD. Statistical analyses were made by using Student’s t-test. P < 0.05 was considered significant.
Table 4. Selected bond lengths and angles for 6chloro-2-(2-naphthyl)-4-phenyl-quinazoline (12).
Bond lengths (Å)
Cl(1)⫺C(5)
N(1)⫺C(1)
N(1)⫺C(8)
C(1)⫺N(2)
C(1)⫺C(15)
N(2)⫺C(2)
C(2)⫺C(3)
C(3)⫺C(4)
C(3)⫺C(8)
C(4)⫺C(5)
C(5)⫺C(6)
C(6)⫺C(7)
C(7)⫺C(8)
C(1)⫺N(1)⫺C(8)
N(1)⫺C(1)⫺N(2)
N(1)⫺C(1)⫺C(15)
N(2)⫺C(1)⫺C(15)
C(2)⫺N(2)⫺C(1)
N(2)⫺C(2)⫺C(3)
N(2)⫺C(2)⫺C(9)
C(3)⫺C(2)⫺C(9)
C(4)⫺C(3)⫺C(8)
C(4)⫺C(3)⫺C(2)
C(8)⫺C(3)⫺C(2)
C(5)⫺C(4)⫺C(3)
C(4)⫺C(5)⫺C(6)
C(4)⫺C(5)⫺Cl(1)
C(6)⫺C(5)⫺Cl(1)
C(7)⫺C(6)⫺C(5)
C(6)⫺C(7)⫺C(8)
N(1)⫺C(8)⫺C(7)
N(1)⫺C(8)⫺C(3)
C(7)⫺C(8)⫺C(3)
Angles (degrees)
1.733(2)
1.324(3)
1.365(3)
1.363(3)
1.488(3)
1.326(2)
1.426(3)
1.413(3)
1.413(3)
1.363(3)
1.403(3)
1.362(3)
1.405(3)
116.64(18)
125.95(19)
117.64(18)
116.39(18)
118.00(18)
121.27(18)
116.01(18)
122.72(18)
118.81(19)
125.16(19)
116.03(19)
119.6(2)
121.8(2)
119.40(17)
118.85(17)
119.3(2)
120.9(2)
118.50(19)
121.96(19)
119.5(2)
 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
246 Mikiciuk-Olasik et al.
collection did not change its appearance. Lorentz polarization
correction was applied to the intensity data. Numerical absorption correction was used [15]. The maximum and minimum transmission factors were 0.609 and 0.656. The structure was solved by direct methods and subsequently completed by the difference Fourier recycling. All the non-hydrogen atoms were refined anisotropically using full-matrix,
least-squares technique on F2. All the hydrogen atoms were
found from difference Fourier synthesis after four cycles of
anisotropic refinement, and refined as “riding” on the adjacent
carbon atom with individual isotropic temperature factor equal
1.2-times the value of the equivalent temperature factor of
the parent carbon atom. The solution and refinements were
performed with SHELXS97 [16] and SHELXL97 [17]. The
graphical manipulations were performed using the XP routine
of the SHELXTL [18] and ORTEP [19]. Atomic scattering factors were those incorporated in the computer programs.
Selected interatomic bond distances and angles are listed in
Table 4.
Experimental details
Chemical formula: C24H15ClN2; formula weight M = 366.83;
crystal system: monoclinoic; unit-cell dimensions a =
10.8004(8) Å, b = 9.8615(9) Å, c = 16.8741(11) Å, β =
95.894(5)° and volume V = 1787.7(2) Å3, temperature 293(2)
K; space group symbol P21/n (alternative setting of P21/c);
number of formula units in unit cell Z = 4, linear absorption
coefficient µ = 0.224 mm⫺1; number of unique reflections
3184 (R(int) = 0.00); final R values: R1 = 0.0455, wR2 =
0.1061 for I>2δ(I) and R1 = 0.0717, wR2 = 0.1223 for all data.
Supporting information
Tables of crystal data and structure refinement, anisotropic
displacement coefficients, atomic coordinates and equivalent
isotropic displacement parameters for non-hydrogen atoms,
H-atom coordinates and isotropic displacement parameters,
bond lengths and interbond angles have been deposited with
the Cambridge Crystallographic Data Centre under
No.CCDC 196439.
References
[1] E. Mikiciuk-Olasik, K. Błaszczak-Świa̧tkiewicz, Wiad.
Chem. 2000, 54, 9⫺10, 705⫺724.
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 239−246
[3] A. Monge, J. A. Palop, J. Med. Chem. 1995, 38,
1786⫺1792.
[4] A. Nunn, K. Linder, H. W. Strauss, Eur. J. Nucl. Med.
1995, 22, 3, 265⫺272.
[5] H. Gnichtel, Chem. Ber. 1970, 103, 411⫺2417.
[6] J. Lessel, Arch. Pharm. 1995, 328, 397⫺402.
[7] E. Mikiciuk-Olasik, K. Błaszczak-Świa̧tkiewicz, Nucl.
Med. Rev. 2000, 3, 2, 149⫺152.
[8] D. Cremer, J. A. Pople, J. Am. Chem. Soc. 1975, 97,
1354⫺1358.
[9] G. R. Desiraju, T. Steiner in The Weak Hydrogen Bond
in Structural Chemistry and Biology, Oxford University
Press, Oxford, 1999.
[10] G. A. Jeffrey, W. Saenger in Hydrogen Bonding in Biological Structures, Springer-Verlag, Berlin, 1994.
[11] R. Taylor, O. Kennard, J. Am. Chem. Soc. 1982, 104,
5063⫺5070.
[12] S. Biniecki, Preparatyka śodków leczniczych,
Państwowy Zakład Wydawnictw Lekarskich, Warszawa
1980, 113.
[13] F. Hoffmann-La Roche & Co. Belg. 616,024. 1962.
[Chem. Abstr. 1963 58. P 10222f].
[14] H. G. Drexler, W. Dirks, R. A. F. MacLoad, H. Quentmeier, K. G. Steube, C. C. Uphoff (Eds.), DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen
catalogue of Human and Animal Cell Lines, Eighth Edition, DZMS, publ., Braunschweig, 2001.
[15] X-RED. Version 1.18. STOE & Cie GmbH, Darmstadt,
Germany, 1999.
[16] G. M. Sheldrick, Acta Cryst. 1990, A46, 467⫺473.
[17] G. M. Sheldrick, SHELXL-97. Program for the Refinement of Crystal Structures, University of Göttingen, Germany, 1997.
[18] G. M. Sheldrick, Release 4.1 of SHELXTL PCTM for
Siemens Crystallographic Systems, Siemens Analytical
X-Ray Instruments Inc., 1990.
[19] L. J. Farrugia, J. Appl. Cryst. 1997, 30, 565.
[2] J. M. Brown, Br. J. Cancer 1993, 67, 1163⫺1170.
 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Документ
Категория
Без категории
Просмотров
0
Размер файла
117 Кб
Теги
oxide, quinazoline, activity, expected, new, derivatives, antitumor, dihydroquinazolinones
1/--страниц
Пожаловаться на содержимое документа