New series of substituted heterocycles derived from α, β-unsaturated ketone and their cytotoxic activity in tumor cell lines

Article abstract of DOI:10.13005/ojc/340620

The aldol condensation of 2-acetylnaphthalene with 9-anthracene carboxaldehyde afforded α, β-unsaturated keton (1) . New heterocyclic compounds containing: cyclohexenone[2], indazole[3], pyrimidinethion [4], thiazolo fused pyrimidine[5], isoxazoline[6], s

Full text of DOI:10.13005/ojc/340620

ISSN: 0970-020 X
CODEN: OJCHEG
2018, Vol. 34, No.(6):
Pg. 2826-2831
ORIENTAL JOURNAL OF CHEMISTRY
An International Open Access, Peer Reviewed Research Journal
www.orientjchem.org
New Series of Substituted Heterocycles Derived from α, β-unsaturated
Ketone and Their Cytotoxic Activity in Tumor Cell Lines
MUNA S. AL-RAwI*, HUDA A. HASSAN* and DHEEFAF F. HASSAN*
Department of Chemistry, College of Education For Pure Science Ibn Al- Haitham, University of Baghdad,
Baghdad- Iraq.
*Corresponding author E-mail: tagreedaloom@gmail.com
http://dx.doi.org/10.13005/ojc/340620
(Received: September 30, 2018; Accepted: November 06, 2018)
AbSTRACT
The aldol condensation of 2-acetylnaphthalene with 9-anthracene carboxaldehyde afforded
α, β-unsaturated keton (1) .New heterocyclic compounds containing:cyclohexenone[2], indazole[3],
pyrimidinethion [4], thiazolo fused pyrimidine[5], isoxazoline[6], substituted pyrazoline[7]a-d and
pyrimidinone[8] rings system were synthesized from α, β-unsaturated keton[1].Cyclization of [1] with
ethylacetoacetate gave the mentioned heterocycle cyclohexanone [2]. The cyclo condensation of
[2] with hydrazine gave the new indazole derivative [3]. furthermore, the reation of [1]with thiourea
gives thiopyrmidine derivative [4].The cyclo condensation of [4] with chloroacetic acid gave the fused
rings [5].Then reacted compound[1] with hydroxylamine to produce isoxazoline [6].Also the reaction
of [1] with hydrazine and hydrazide derivatives to produce pyrazole [7]a-d. All the newly synthesized
1
derivatives[2-7a-d] were characterized via the CHN-S, FT- IR, H NMR, and 13C NMR (of some of
theme). The antibacterial and cytotoxic activities were evaluated for these derivatives[2-7a-d]. The
study of antibacterial displayed good to moderate activity and the study of anticancer activity showed
that were effective for inhibition of L20B the mice intestines carcinoma cell line and RD human pelvic
rhabdomyosarcoma cell line. treated
Keywords: α, β-Unsaturated ketons, Cyclohexenone , Indazole, Pyrimidinethion, Isoxazoline,
Pyrazoline, Pyrmidinone, Anti-cancer activity.
INTRODUCTION
pyrimidinethion, isoxazoline, pyrazoline, and
pyrimidinone3. The biological activity for chalcones
are wide ranging in the recent years, isoxazolines
are considered biologically active molecules since
they are used in controlling parasite infections in
humans4 in addition to their use as insecticides5,
nematicides and molluscicides agents6.Pyrazolines,
for example, have attracted increasing attention
α,β–Unsaturated ketons, one of the
primary classes of natural products and belongs
to flavonoid family, as well these compounds keep
several biological activities1,2. α, β-Unsaturated
ketons are suitable for the synthesis of an important
heterocycles rings like cyclohexenone, indazole,
This is an
Open Access article licensed under a Creative Commons license: Attribution 4.0 International (CC- BY).
Published by Oriental Scientific Publishing Company © 2018

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AL-RAwI et al., Orient. J. Chem., Vol. 34(6), 2826-2831 (2018)
due to their pharmaceutical applications such as:
General Synthetic procedures
All compounds [2-8] were synthesized
confer to the scheme 1.
anti-bacterial, anti-fungal, enzymatic inhibitors
and cytotoxic properties7-9. Pyrmidine derivatives
play an essential role in medicinal field due to their
anti-inflammatory, anti-ulcerogenic10. The anti-
inflammatory and anti-cancer activities of indazoles
were also reported11. Hence, it appeared of interest
to prepare the mentioned heterocycles linked to
2-acetyl naphthalene moiety and evaluated their
cytotoxicity effect on two cancer cell lines including:
L20B and RD cell lines.
preparation of of chalcone:3-(anthracen-9-yl)-1-
(naphthalen-2 -yl) prop–2–en-1-one[1]
2-Acetyl naphthalene (1.70 g, 0.01 mol)
was added with small portions to a solution of
(2.06 g, 0.01 mol) 9-anthracenecarboxaldehyde
in 10% ethanolic NaOH (10 mL) and stirred for
3 hours. The reaction mixture was acidification with
dilute HCl. The resulting product was filtered off,
washed well with cold water, dried in air to give the
required product [1]; Yield 82%; m.p 155-157oC;
FT- IR : 3049 (C-H arom.), 2931 ( C-H aliph.), 1658
(C=O), 1622- 1597 (C=C) and 1172due to ( C-O-C);
1H-NMR (δ):8.42-7.70(m,16H,Ar-H),6.20,6.15(2H,d,
CH=CH). Furthermore, the results of CHN-S analysis
harmonized with the proposed structure for [1] C27H18O:
C, 90.47; H, 5.06; Found:C, 90.91; H,5.63.
ExpERIMENTAL
Materials
All chemicals utilized brand Sigma-Aldrich
and used as received.
Instrumentation
FTIR spectra were registered in KBR dices
on a SHIMADZU-FTIR-8400spectro photometer.1H
&
13CNMR spectra were done on Bruker 400-MHz
spector photometer. CHN-S were completed an
EuroEA Elemental Analyzser.
Ar
Ar
Ar
Ar
O
N
O
O
O
NH
N
O
NH
ClCH2COOH
NH2NH2
Ar
Ar
N
H
S
Ar
N
S
Ar
[2]
[3]
[4]
[5]
C
H
3 C
O
C
H 2
H
2 C
N
S
O
C
O
2
H
E
N
t
Ar
O
Ar
[1]
N
H
2 C
l
C
O
N
H
.
H
H
2
O
H 2
N
Ar
Ar
Ar
R
O
N
N
HN
N
6]
[8]
[
N
H
Ar
O
[7]a-d
Ar
Ar
H, ph , -CO-2-NH2C6H4
, NH-CS -NH2
R =
Ar =
/
Ar =
Scheme 1. Synthetic route for target derivatives [1-8]

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AL-RAwI et al., Orient. J. Chem., Vol. 34(6), 2826-2831 (2018)
Synthesisof ethyl6-(anthracen-9-yl)-4-(naphthalen-
2-yl)-2-oxocyclohexa-3-ene-carboxylate [2]
Synthesisof5-(Anthracen-9-yl)-7-(naphthalen-2-yl)
-5H- Thiazolo [3,2-a] pyrimidin-3(2H)- one [5]
Chloroacetic acid (0.01 mol) was added
A stirred mixture of chalcone [1] (3.58 g,
0.01 mol), CH3COOC2H5 (0.01 mol) ,aqueous KOH
solution (1mL, 10%) was heated for 3 h and then
left to stirring overnight at ambient temperature.
The precipitated off white was collected and dried
to give the cyclohexanone derivatives [2]; Yield :
68% ;m.p 182-1840C;FTIR 3035 (C-H arom.), 2981
( C-H aliph.), 1737 (C= O) 1660 (C =O) and 1604
(C =C);1HNMR:8.55-6.92 (m, 16H, H-Ar ), 6.69(s,1H,
CH=C-Ar),4.02(1H,m,CH-Ar/ ), 2.71 ( 1H, s, CH-CO),
1.58 (s, 2H, CH2-C-Ar), 1.24 (q,2H, -CH2CH3) and
1.10 (t, 3H, -CH2CH3); Anal. Calcd. for C33H26O3:
C( 84. 23); H(5.57); Found:C(84.85); H (5.76).
to a compound[4] (4.16 g,0.01 mol) in mixture of
glacial CH3CO2H (10 ml) and (CH3CO)2O (5ml) and
anhydrous sodium acetate (0.45 g). The mixture
refluxed for 6 h and poured into 30 mL crushed ice
to allow golden yellow solid precipitate, which was
filtered and dried; Yield ( 48% ); M. P. 193-1950C;
FT-IR (ν, cm-1): 3091 (C-Haromatic), 2098 (C-H
aliphatic), 1668 (C=O),1606 (C=N), 1078(C-S);
1H NMR (9.55-7.16:m, 16H Ar-H), 8.46 (d, 1H,
CH=CAr ), 5.45 (d,1H, =CH-CH-Ar), 1.66(s,2H,CH2).
CHN-S for compound C30H20N2OS: (C, 78.92);
(H, 4.42); (N, 6.14); (S, 7.02); Found: (C,79.20);
(H,4.18); (N,6.45); (S,7.42).
Synthesis of 4-(anthracen-9-yl)-6-(naphthalen-2-
yl) -2 ,3 ,4 ,5 -tetrahydro-3H-indazol-3-one [3]
Hydrazine hydrate was added dropwise
Synthesis of derivatives 5-(anthracen-9-yl)–3-
(naphthalene–2–y l )-4 , 5-dihydro -isoxazole[6]
To a mixture of chalcone[1] (3.58 g, 0.01
to a solution of compound[2] (4.70 g,0.01 mol)
in ethanol (20 mL) and glacial acetic acid (0.3
mL ) at ambient temperature. The mixture was
heated for 6 h12. The pale yellow solid formed was
filtered off and dried, Yield:52%; M. P. 137-1390C; FT-
IR(3385-NH), (3020C-H arom.), (2980 C-H aliph.),
(1653 C = O) (1631 C = N) and (1955 C=C); 1HNMR
(9.78:s,1H,NH), (7.86-7.22 : m, 16H, H-Ar ), (4.02:s,
1H, CH =C-Ar), (3.47 :1H, d, CH-CO), 3.18(d, 2H,
CH2CH-Ar/ ), 1.35 (m, 1H, CH2-CH- Ar/); likewise, the
results of CHN-S analysis agree with the suggested
structure for compound(3) :C31H22N2O: (C:84.91);
(H:5.06); (N:6.39) ; Found:(C: 85.15); (H:5.66,
(N:6.91).
mol) and hydroxyl amine hydrochloride (0.69 g,
0.01 mol) ethanolic sodium hydroxide (10 mL)
was refluxed for 8 hours. On cooling 50 mL water
was added then the mixture acerbated at fridge
overnight and the white solid was formed, Yield
(77% ); m.p. 146-1480C; FT-IR3064 (C-H arom.),
1627 (C=N), 1598 (C=C);1348 (cyclic ether) (C - O);
1HNMR:(8.89-7.36:m, 16H, ArH), (4.73-4.61 :d, 2H,
CH2CHAr/), (2.25-1.20 :t,1H, CH2-CH- Ar/).The CHN
analysis for C27H19NO:(C, 86.84);(H, 5.13);(N, 3.75);
Found: (C,86.22); (H,5.45); (N,3.95).
Synthesis of 5-( naphthalen -2-yl)-3- (substituted
phenyl) - pyrazoline derivatives [7]a-d
A mixture of chalcon [1] (3.58 g, 0.01 mol)
and hydrazine hydrate or substituted hydrazine (0.01
mol) in EtOH (20 mL) containing glacial acetic acid
(0.3 mL) was heated for 4 hour. The precipitated
was formed , collected and dried to afford [7]a-d.
Synthesis of 4-(anthracen-9-yl)-6-(naphthalen-2
-yl)-3, 4-dihydropyrimidine 2 (1H) -thione [4 ]
To a solution of chalcone (3.58 g) in
ethanolic NaOH (0.01 mol), SC(NH2)2 (0.6 g) was
added, refluxed for 7 hours. A solution of ice cold
water was added, the orange solid created, washed
with water and dried, yield :68%; m.p. 214-2160C ;
FT-IR: 3383 cm-1 for(NH), 3012 cm-1 (C-H arom.),
5-(anthracen-9-yl)-3-(naphthalen-2-yl)-4,5-
dihydro-1H-pyrazole[7]a
Yellowish brown;Yield (58% );m.p. 92-940C;
FTIR for this compound confirm that: 3431(NH),
3053 (CH -Ar), 1666 (C =N), 1593 (C =C); 1HNMR
:(9.01:s,1H, NH), (8.48 -6.76: m, 16H Ar-H), 4.11
(t,1H, CH2CHAr/ ), 2.49-1.46 (d,2H, CH2-CH- Ar/),
CHN analysis (C27H20N2) agree with the proposed
structure:C, 87.07;H, 5.41;N,7.52 Founder:C,87.55;
H,5.85; N,7.85.
1
1638 (C =N), and 1255 (C =S). HNMR: 12.00
(s,1H, NH tautomeric with SH in thiopyrimidine
ring), 11.38(s,1H,NH),9.00-7.86(m, 16H ArH), 8.29
(d ,1H, CH = C-Ar ), 7.65 (d ,1H, = CH-CH-Ar); Also,
the output of CHN-S analysis correspond with the
indicate structure for C28H20N2S.

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AL-RAwI et al., Orient. J. Chem., Vol. 34(6), 2826-2831 (2018)
5-(anthracen-9-yl) -3- (naphthalen-2 –yl ) -1- phenyl-4,
5- dihydro -1H-pyrazole[7]b
Dark orange;Yield ( 60% );m.p.149-1510C;
FTIR(CH-Ar 3057), (C =N1620);1HNMR, 8.12 -7.23
(m, 16H ArH), 3.67 (t,1H, CH2CHAr/ ), 1.19-1.35
(d,2H, CH2-CH- Ar/); the CHN-S compound[7]b
C33H24N:C,88.36;H,5.39;N,6.25, Founder:C,88.99;
H,5.70; N,6.05.
RESULTS AND DISCUSSION
Aldole condensation reaction of 2-acetyl
naphthalene with 9-anthracenecarboxaldehyde in
alcoholic sodium hydroxide offorded3-(anthracen
-9-yl)-1-(naphthalen-2-yl) prop-2-en-1-one[1] at
room temperature.The structural of the chalcone[1]
was substantiated spectroscopically and elemental
analyses .The formation of cyclohexanone derivative
[2] was achieved by refluxing chalcone[1] with
ethylacetoacetate in the presence of aqueous
potassium hydroxide solution. Moreover, reaction
of this derivative[2] with hydrazine hydrate in the
presence of CH3CO2Hasacatalystgiveanewindazol
derivative[3], the reaction may have proceeded
through condensation between carbonyl group
of cyclohexanone[2] and amineNH2 of hydrazine,
followed by cyclization by losing a molecule of
ethanol. The pyrimidinethione derivative [4] was
obtained from refluxing chalcone [1] with thiourea
in basic medium, its structure was ascertained by
spectral data.However, compound [5] was prepared
directly from the reaction [4] with ClCH2CO2H in
glacial CH3CO2Hand (CH3CO)2O in the presence of
anhydrous CH3CO2Na. Isoxazoline compound [6]
was synthesized from the reaction of chalcone [1]
with NH2OH.HCl. Sutructure of compound[6] was
established by their spectral data, the FTIR spectra
were no characteristic bands of the CH=CH and
C=O and NH groups in the starting material, with
the appearance of new absorption band for (C=N)
group around 1627 cm-1 and C-O (cyclic ether) group
around 1348 cm-1. It was reported that, reaction of
α, β-Unsaturated ketone with N2H4 or PhNHNH2
or 2-amino benzohydrazide or thiosemicarbazide
in the presence of glacial acetic acid and refluxed,
yield the target pyrazoline [7]a-d, respectively. The
structure of the pyrazoline derivatives [7]a-d were
identified by their melting point, CHN-S analysis,
FTIR and 1HNMR spectroscopy. The FTIR spectra
of these compounds exhibited new absorption
stretching peaks of NH and imiene groups in the
region 3431cm-1 and 1666-1660 cm-1, the more
characteristic for pyrazole ring. The pyrimidinone
derivative [8] was synthesized from reaction of
chalcone [1] with urea in glacial acetic acid as
a catalyst. The structure of the pyrimidinone [8]
characteristic by FTIR, H & 13C NMR spectroscopy;
the suggested mechanism of this reaction may be
shown as follows Scheme 2.
(2-aminophenyl)(5-(anthracen-9-yl)-3-(naphthalen-2-
yl) -4 , 5- dihydro-1H-pyrazol -1- yl) methanone [7]c
Brownish red;Yield (74%);m.p.172-1740C;
FTIR: 3344(NH2), 3026 (CH -Ar), 1666 (C =N),
1593 (C =C), 1336 (C=S); 1HNMR : 9.32(s,1H,NH),
8.70 -7.55 (m, 16H, Ar-H), 4.27 (t,1H, CH2CHAr/ ),
1.711.21 (d,2H, CH2-CH- Ar/);CHN-S for compound
C34H25N3O [7]c:C, 83.07;H, 5.13;N, 8.55;Found:C,
83.65; H, 5.42; N,8.76.
1-(5-(anthracen-9-yl)-3-(naphthalen-2-yl)-4,5-
dihydro -1H- pyrazol -1 -yl) thiourea [7]d
Bright yellow; Yield (67%); m.p. 204-
2060C; FT-IR(ν, cm-1): 3309(NH2), 3035 (C-H
aromatic),1718(C=O), 1660 (C=N), 1597 (C =C);
1HNMR: 11.47(s,1H,NH2), 9.70 -7.45 (m, 16H
Ar-H), 7.60-7.61 (t,1H, CH2CHAr/), 1.62 (d,2H, CH2-
CH- Ar/); Anal. Calcd. for C28H22N4S: C, 75.31; H,
4.97; N, 12.55; S, 7.18 Founder: C, 75.73; H, 5.11;
N,12.32;S,7.29.
Synthesis of 4-( anthracen-9–yl )-6-( naphthalene
-2 -yl)-3,4-dihydropyrimidin-2(1H)-one [8]
To a solution of chalcone[1] (3.58 g,
0.01 mol), urea (0.6 g, 0.01 mol) and EtOH (30 mL)
containing glacial CH3CO2H (0.3 ml) was added.
The combination was heated for 7 h and poured
into crushed ice. The solid light yellow precipitate
was washed, filtered and dried to give pyrimidinone
derivative[8]. Yield (75%); m.p. 119-1210C ; FTIR:
(3406, 3350NH), (3064 CH arom.), (1670 C=O),
1
(1614,1597 C=C); HNMR: 10.12 (s,1H, NH ),
8.82(s,1H, NH), 8.38-7.35 (m,16H, Ar-H and
pyrimidine proton ), 3.33 (d,1H, =CH-CH ); 13CNMR
the presence of peaks at δ 190.88 (C=O), δ 125.11-
138.96 (C aromatic) and δ 123(CH=CH) ; CHNS for
C28H20N2O the target compound: C, 83.98; H, 5.03;
N, 7.00; Found: C,83.67; H,5.43; N,7.11.

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O
O
NH2 - C - NH
NH2 - C - NH2
+ NaOH
Ar
Ar
Ar
N
O
proton
transfer
NH2 - C - NH
O
OH
Ar
+
NH2
NH2
Ar
O
C
O
HN
Ar
C
O
Ar
Ar
Ar
N
HN
C
H
H2O
HN
C
CH
OH
N
O
NH
Ar
(E)
C
O
O
N
Ar
Ar
Ar
HN
C
Ar =
N
H
O
Ar
/
Ar =
Scheme 2
Antibacterial evaluation
24 hours.(+) and (+ +) depending upon the diameter
and clarity as inTable 1.The derivatives [3], [5], [7]b,
[7]d, and [8] displayed good to moderate antibacterial
activity against three pathogenic species; also, the
derivatives [2], [6], [7]a and [7]c shows low activity
against Gram positive. Finally, both compounds[2]
and [7]c shows no activity against Gram negative
but they exhibited moderate to weak antibacterial
against Gram poistive.
The target compounds were synthesized
[2-8] were screened their antibacterial activity
(in vitro) against both G+ and G- bacteria;like Staph.
aureus(G+), Bascillus cereus (G+) and E.coli(G-)
according to the agar plate diffusion method. The
test solution was prepared by using DMSO as a
solvent, and each compounds was dissolved in
DMSO to give concentration (1 ppm). The plates
were then incubated at 370C and examined after
Table 1 : Antibacterial activity of the synthesized compounds[2-8]
Comp. Staphlococcus Bascillus cereus E. coli
Comp. Staphlococcus Bascillus cereus E. coli
No.
aureus(G+)
(G+)
(G-)
No.
aureus(G+)
(G+)
(G-)
[2]
[3]
[4]
[5]
[6]
7.1
20.4
11.7
18.7
8.1
12
16.8
6.6
13
0
[7]a
[7]b
[7]c
[7]d
[8]
5.5
27
10
19.2
13.7
15
7.7
7.5
11
6.9
9.8
0
8.8
6.7
6.5
13.7
11.9
8
9.5
11.8
Key to symbols: highly active = ≤15 mm, moderately active =11-15 mm and slightly active =5-10mm.
Cytotoxic activity
All derivatives of chalcone were chosen for
well from single cell hang were added to all the
96 wells of the microtiter plates ; which wrapped by
plate lids and sealed with parafilm, shaked softly
and returned to the incubator.
examined their anticancer activity.Two cell lines were
used for testing: L20B (mice intestines carcinoma)
cell line and RD (human pelvic rhabdomyosarcoma)
according to the procedure of Freshney (13). Seven
derivatives [2-8] different concentrations were
tested for their cytotoxicity by using cultured cells in
microtiter plate (96 wells) .The examine was applied
by the next steps:
b- Incubation: Plates were incubated in moisten
chamber at 37oC, 5% CO2.
c- Exposure: when the cells are in full activity, they
were exposed to 6 concentrations of the derivatives
µg/ml for cell line, 200µl of maintenance medium
were added to each well of control group, then
a-Seeding: when cells in the incubated falcon
became monolayer, a liquot 200 µl/104-105 cells/

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plates were locked with parrafilm and returned to
the incubator. cytotoxicity was carried out after 48 h
and the photo picture were taken after 24 hour.
The synthesized derivatives [5], [7]a, [7]
b,[7]c and [8] showed moderate anticancer activity
against two type of cancer cell lines.The remaining
derivatives exhibited low anticancer activity.However,
the compound [6] showed no activity versus both cell
lines, and [2] showed no activity versus RD cell line.
Structure-activity relationship could be observed by
examining the effect of heterocyclic rings on these
derivatives on the cell growth inhibition.
d- Staining: Cell viability was measured after 48 h,
then removing the medium and 20 µl/well solution
of MTT were added. The crystals remaining in the
wells were solubilized by the addition of 200 µl/
well of DMSO followed by incubation in 37°C for
15 min.with shaking.The absorbance was measured
on a microplate reader at 620 nm .The average of
inhibition of cell growth was counted according to
(46) as follow equation.
Table 2 :The inhibition of cells growth of synthesized derivatives. µl/well
Comp. No. For (L20 B) For (RD) Comp. No. For (L20 B) For (RD)
[2]
[3]
[4]
[5]
[6]
20.00%
15.90%
11.50%
52.70%
-
-
[7]
47.80%
60.70%
18%
51.90%
23%
42.10%
33.50%
11.90%
32.30%
34.20%
23.30%
25.70%
12.20%
-
[7]a
[7]b
[7]dc
[8]
CONCLUSION
The study of antibacterial displayed good to moderate
activity. All compounds except [6] showed moderate
to low inhibition for L20B and RD cancer cell lines.
The aim of the present work has been
directed towards synthesized of new series of
heterocyclic compounds derived from α, β-un
saturated ketone they may be have more activity
and less toxicity as anticancer agents. Biological
evaluation may support the development of synthetic
organic compoundes and pharmaceutical chemistry.
ACKNOwLEDGMENT
we thank Dr. Hytham Ahmed -Faculty of
Pharmacy -Menoufia University/ Egypt for support
us throughout our experimental work.
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