Microwave-assisted synthesis of certain pyrrolylpyridines, some derived ring systems and their evaluation as anticancer and antioxidant agents

Article abstract of DOI:10.1016/j.ejmech.2015.01.023

The synthesis of 18 novel pyrrolylpyridines and some derived bi-, tri- and tetracyclic ring systems using both the conventional heating and MW irradiation techniques is described. Fourteen compounds; 2-9, 10-12, 14, 17 and 18 were evaluated for their anti

Full text of DOI:10.1016/j.ejmech.2015.01.023

Accepted Manuscript
Microwave-Assisted Synthesis of Certain Pyrrolylpyridines, Some derived Ring
Systems and Their Evaluation as Anticancer and Antioxidant Agents
Sherif A.F. Rostom, Adnan A. Bekhit
PII:
S0223-5234(15)00043-4
10.1016/j.ejmech.2015.01.023
EJMECH 7639
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 18 September 2014
Revised Date: 10 January 2015
Accepted Date: 11 January 2015
Please cite this article as: S.A.F. Rostom, A.A. Bekhit, Microwave-Assisted Synthesis of Certain
Pyrrolylpyridines, Some derived Ring Systems and Their Evaluation as Anticancer and Antioxidant
Agents, European Journal of Medicinal Chemistry (2015), doi: 10.1016/j.ejmech.2015.01.023.
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ACCEPTED MANUSCRIPT
Microwave-Assisted Synthesis of Certain Pyrrolylpyridines, Some derived Ring Systems
and Their Evaluation as Anticancer and Antioxidant Agents.
Sherif A. F. Rostom*, Adnan A. Bekhit
Several pyrrolylpyridines (A) and some derived fused rings (B-D) were synthesized using both
conventional and MW heating techniques. Some pyridines, tetrahydroquinolines. and
tetrahydropyrimido[4,5-b]quinolines exhibited promising antitumor and antioxidant activities.

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Microwave-Assisted Synthesis of Certain Pyrrolylpyridines, Some derived
Ring Systems and Their Evaluation as Anticancer and Antioxidant Agents.
Sherif A. F. Rostom,*1,2 Adnan A. Bekhit2
1 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, King Abdulaziz University, P.O. Box
80260, 21589 Jeddah, Saudi Arabia
2 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Alexandria, Alexandria
21521, Egypt
* Corresponding Author: Tel: +966 507654566; Email: sherifrostom@yahoo.com
Abstract
The synthesis of 18 novel pyrrolylpyridines and some derived bi-, tri- and tetracyclic ring
systems using both the conventional heating and MW irradiation techniques is described.
Fourteen compounds; 2-9, 10-12, 14, 17 and 18 were evaluated for their antitumor activity
according to the National Cancer Institute (NCI), in vitro disease oriented antitumor screening.
Distinctive antitumor activity was conjugated with compounds 3 and 7 (R = 3,4-di-OCH3-C6H3).
The analogs 3, 6, 7, 9, 10, 11 and 12 which exhibited prominent antitumor activity, were further
evaluated for their antioxidant potential using the DPPH radical scavenging assay. The
substituted 6-(3,4-dimethoxyphenyl)pyridine-3-carbonitriles 3 and 7 were nearly equipotent to
BHT the standard antioxidant utilized in this assay (scavenging activity 31 and 33 %,
respectively, vs 36 %). Accordingly, compounds 3 and 7 can be considered as lead structures for
dual antitumor and antioxidant activities.
Keywords:
Pyridines, Tetrahydroquinolines, Pyrimido[4,5-b]quinolines, Antitumor activity, Antioxidant
activity.

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1. Introduction
Despite the persistent efforts and prominent accomplishments achieved in many domains of
cancer treatment, effective control of this complicated disease continues to be a major quarrel for
medicinal chemistry researchers. This aspect refers to the fact that cancer is a complex
pathological condition with a diversity of manifestations distinguished by rapid proliferation and
spreading of anomalous cells [1]. To date, therapeutic management of cancer is a conjunction of
surgery, radiotherapy together with chemotherapy, which remains one of the most reliable tools
in the control and even recovery from different types of tumors. However, administration of
anticancer drugs usually accompanied by several complications among which toxicity of healthy
cells and the possible evolution of drug resistance [2]. Consequently, the need for designing
novel unparalleled antitumor agents becomes an urgent demand.
On the other hand, oxidative stress caused by the prevalence of high levels of reactive oxygen
species (ROS) such as hydroxyl free radical and superoxide radical anion that drastically attack
and damage cells throughout the body [3], has been linked with the etiology of a variety of
pathophysiological conditions such as rheumatoid arthritis, autoimmune diseases and definitely
cancer [4]. Moreover, ROS was found to induce oxidative damage of DNA, which could result in
mutations leading to carcinogenesis. Besides, ROS were shown to enhance tumor proliferation
and progression through activation of signaling pathways involved in angiogenesis and
metastasis [5]. Therefore, antioxidants that are able to scavenge free radicals and keep cell
integrity have become currently an issue of growing interest, and their therapeutic utility would
contribute significantly in the management of several types of cancer [6,7].
Among the diverse classes of heterocyclic compounds that are endowed with multiple biological
efficiencies, pyridines and their derived fused ring structures have received profound attention
since they are proved to be biological versatile compounds. A wide range of chemotherapeutic
activities have been ascribed to pyridine derivatives including antimicrobial [8,9], antitubercular
[10] and anticancer [11-15], and antiviral [16] activities. In particular, interest has been focused
on pyridinecarbonitriles as they possess significant antibacterial [17], antifungal [18] and
anticancer activities [19-21], in addition to their well-documented antioxidant potentials [22,23].
Consequently, pyridinecarbonitriles and their derived fused rings remain a promising template
for the design of a new category of chemotherapeutic and antioxidant agents.

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Along the past two decades, the utilization of microwave (MW) irradiation as a substitute of
other conventional heating techniques has acquired popularity in contemporary synthetic
medicinal chemistry research. Successful implementation of MW irradiation in various chemical
reactions resulted in enhanced reaction rates with subsequent time-saving, fast and effective
building up of libraries of compounds, improvement in products’ purity and yields, in addition to
being a suitable route towards the goal of echo-friendship and green chemistry [24].
[Figure 1]
Inspired by the aforementioned findings, and as an extension to our efforts devoted to the
preparation of heterocycles with powerful chemotherapeutic potentials [25-43], it was attempted
to synthesize some novel pyridines and some derived bi-, tri- and tetracyclic ring systems having
the general structures (A-D; Fig. 1) using the MW irradiation technique, to be biologically
evaluated as anticancer and antioxidant agents. Seeking synergistic biological efficiency, the
targeted compounds are aimed to encounter basically a 1-methylpyrrolyl moiety owing to the
reported role of the pyrrole ring in inducing many anticancer [44,45] and/or antioxidant [46,47]
activities. In addition, the target compounds are supported with several functionalities that would
assist the aimed biological actions. Therefore, this research work points to achieve new
molecules that would be implemented as pioneers in the anticancer and/or antioxidant drug
discovery area.
2. Results and Discussion
2.1. Chemistry
The pathways adopted for the preparation of the intermediate and target compounds are depicted
in schemes 1 and 2. The reactions employed in these schemes were studied both under
conventional heating conditions (fusion or reflux in the appropriate solvent), as well as single
mode microwave (MW) irradiation technology. Optimization of the conditions of the performed
reactions was accomplished via employment of various combinations of temperature and
reaction time to achieve the maximum chemical yield at the shortest reaction time (TLC
monitoring). The documented high heating efficiency of MW resulted in an observable

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acceleration in reaction rates as revealed from the dramatic reduction in reaction times (from 0.5-
8 hours to 10-20 minutes) and the observable improvement in reaction yields (Table 1).
[Scheme 1]
In scheme 1, the 4,6-disubstituted-2-oxo-1,2-dihydropyridine-3-carbonitriles 2-4 and the 2-oxo-
1,2,5,6,7,8-hexahydroquinoline-3-carbonitrile 5 were prepared using Hantzsch-type synthesis via
multi-component one-put cyclocondensation of 1-methyl-1H-pyrrole-2-carbaldehyde 1 with the
appropriate ketone, ethyl cyanoacetate and ammonium acetate as a nitrogen source. In a similar
fashion, when the aldehyde 1 was condensed with the appropriate ketone, malononitrile and
ammonium acetate, the analogous 2-amino derivatives 6-9 were obtained. The latter
pyrrolylhexahydroquinoline 9 was reacted with benzoyl isothiocyanate in the presence of K2CO3
in acetone to afford the corresponding thioureido derivative 10. The IR spectrum of 10 revealed
the characteristic CN absorption at 2220 cm-1, in addition to the C=O and C=S bands at 1710 and
1170 cm-1 , respectively. Furthermore, direct condensation of the 9 with thiourea was attempted
to produce the target pyrimidoquinoline-2-thione 11. Its IR spectrum lacked the CN absorption,
however it showed new absorption band at 1180 cm-1 attributed to the (C=S) group. A schematic
presentation of the reaction mechanism involved in the synthesis of compounds 11 is illustrated
in figure 2.
[Figure 2]
The synthesis of the the targeted alkylthio derivatives 12 and 13 was successfully achieved via
two different approaches. The first method involved thioakylation of the 2-thione derivative 11
with the appropriate alkyl halide in 1N NaOH medium using conventional heating procedure.
Whereas, MW irradiation of a mixture of the thioureido derivative 10, the appropriate alkyl
halide and 1N NaOH was utilized as a second method to produce the same alkylthio derivatives
in shorter time and better yields.
[Scheme 2]
In scheme 2, the pyrrolylhexahydroquinoline 9 was utilized as a precursor for the preparation of
the target compounds 14-19. In this respect, reacting 9 with the phenyl isothiocyanate in a
medium of pyridine gave rise to the substituted N-phenyl pyrimidoquinoline-2-thione 14. Its IR
spectrum exhibited a new band at 1635 cm-1 due to the (C=N) moiety and another one at 1210
cm-1 corresponding to the (C=S) group. Moreover, heating 9 with acetic anhydride resulted in the
targeted 2-methyl pyrimidoquinoline analog 15. The IR of 15 revealed a characteristic (C=O)

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band at 1715 cm-1, while its 1H-NMR spectrum showed a new singlet at δ 2.56 ppm attributed to
the newly introduced CH3 group at C2. On the other hand, reacting 9 with triethyl orthoformate
yielded the 2-ethoxymethylidineamino derivative 16. The IR spectrum of 16 showed in addition
to the cyano group absorption at 2223 cm-1, a new band at 1620 cm-1 due to the C=N moiety.
1
Meanwhile, its H-NMR exhibited a characteristic triplet and quartet at δ 1.25 and 3.69 ppm,
respectively, owing to the CH2CH3 group, in addition to the CH=N singlet at δ 8.11 ppm. In its
turn, compound 16 was reacted with NH2NH2.H2O in order to obtain the targeted tricyclic
pyrimido[4,5-b]quinoline analog 17. Its 1H-NMR showed two new singlets at δ 5.1 and 7.91 ppm
due to the novel NH2 and NH protons, respectively. In its turn, compound 17 was subjected to
cyclization reaction either with formic acid to afford the targeted tetracyclic pyrrolyl
tetrahydropentaazacyclopenta[a]anthracene 18, or acetic anhydride to give the corresponding 2-
1
methyl analog 19. The H-NMR of 19 was characterised by the disappearance of the singlets of
NH protons, and the appearance of a new singlet at 2.55 ppm attributed to the C2-CH3 group.
2.2. In vitro antitumor Screening
2.2.1. Preliminary in vitro one-dose antitumor screening
Out of the newly synthesized eighteen analogs, fourteen compounds namely: 2-12, 14, 17 and 18
were selected by the National Cancer Institute (NCI) in-vitro disease-oriented human cells
screening panel assay to be evaluated for their in-vitro antitumor activity. An effective one-dose
assay has been added to the NCI-60 cell screen in order to increase compound throughput and
reduce data-turnaround time to suppliers while maintaining efficient identification of active
compounds [48-50]. All compounds submitted to the NCI-60 cell screen are tested initially at a
single high dose (10 µM) in the full NCI-60 cell panel including leukemia, non-small cell lung,
colon, CNS melanoma, ovarian, renal, prostate, and breast cancer cell lines. For compounds 10-
12, 14, 17 and 18 data are reported as a mean graph of the percent growth of treated cells, and
presented as percentage growth inhibition (GI %) caused by the test compounds (Table 2). The
analogs 2-9 that have successfully met threshold inhibition criteria determined by the
Development Therapeutic Program (DTP), passed this primary anticancer assay, and were
carried over to the 5-dose screen against a panel of about 60 different tumor cell lines.

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The results recorded in table 2 showed that, some of the tested subpanel tumor cell lines
displayed certain sensitivity profiles against most of the tested analogs 10-12, 14, 17 and 18.
Among these, the non-small cell lung cancer EKVX, HOP-92 and NCI-H522 cell lines exhibited
a wide range of sensitivity towards all the tested compounds, with GI range 10.1-36.4%.
Moreover, an observable activity with respect to breast cancer T-47D cell line was displayed by
the analogs 10, 11 and 12 (GI values 40.6, 40.1 and 54.2%, respectively). Meanwhile, the
ovarian cancer IGROV1 cell line showed mild to moderate sensitivity towards all the tested
compounds, with special behavior towards compounds 10-12 with GI values range 34.4-38.8 %.
Concerning the leukemia subpanel, most of the tested compounds revealed considerable growth
inhibitory activities against most of the tested cell lines, especially the RPMI-8226 cell line.
Particular high activities against this cell line were shown by compounds 11, 12 and 14 (GI
values 51.9, 68.5 and 47.6%, respectively), whereas, the analogs 10 and 17 exerted nearly
equipotent growth inhibition on the same cell line (GI values 36.7 and 38.6%, respectively).
Furthermore, a reasonable sensitivity profile towards the tested compounds was shown by the
leukemia MOLT-4 cell line, especially compound 12 (GI value 42.3%). Additionally, the rest of
leukemia subpanel cell lines were moderately affected by the presence of the tested compounds
with GI range of 8.5-35.9 %. Regarding the renal cancer subpanel, compounds 10-12 were able
to remarkably inhibit the growth of the UO-31 cell line with GI values 51.4, 34.7 and 45.8%,
respectively. Colon, melanoma, prostate and CNS cancer subpanels were the least affected by the
presence of test compounds (GI range 6.3-31.9%), except the prostate PC-3 cancer cell line
which showed remarkable growth inhibition with the analog 12 (GI value 57.1%).
2.2.2. In vitro full panel (five-dose) 60-cell line assay for compounds 2-9
Eight compounds 2-9 which passed the primary one-dose screening, were selected to be
evaluated for their antitumor activities according to the in vitro full panel (five-dose) 60-cell line
assay [48-50]. About 60 cell lines of nine tumor subpanels, including leukemia, non-small cell
lung, colon, CNS, melanoma, ovarian, renal, prostate and breast cancer cell lines, were incubated
with five concentrations (0.01-100 ꢀM) for each compound and were used to create log
concentration-% growth inhibition curves. Three response parameters (GI50, TGI, and LC50)
were calculated for each cell line. The GI50 value (growth inhibitory activity) corresponds to the

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concentration of the compounds causing 50% decrease in net cell growth, the TGI value
(cytostatic activity) is the concentration of the compounds resulting in total growth inhibition and
the LC50 value (cytotoxic activity) is the concentration of the compounds causing net 50% loss of
initial cells at the end of the incubation period (48h). Subpanel and full panel mean-graph
midpoint values (MG-MID) for certain agents are the average of individual real and default GI50,
TGI, or LC50 values of all cell lines in the subpanel or the full panel, respectively. The NCI
antitumor drug discovery was designed to distinguish between broad spectrum antitumor
compounds and tumor or subpanel-selective agents. In this part of the present study, compounds
2-9 exhibited a wide range of antitumor activities against some of the tested subpanel tumor cell
lines (GI50, TGI and LC50 values <100 ꢀM). These compounds showed particular sensitivity
towards some individual cell lines, as well as a broad spectrum (MG-MID) of antitumor activity
(Table 3).
With respect to the sensitivity against some individual cell lines, within the substituted 2-
oxopyridine-3-carbonitriles 2-4, compound 2 (R = C6H5) revealed mild to moderate growth
inhibitory activity against 8 tumor cell lines belonging to 6 different subpanels (GI50 range 27.5-
97.7 ꢀM), with exceptional impact on the non-small cell lung Hop-92 cell line at both the growth
inhibitory (GI50) and cytostatic (TGI) levels (0.11 and 0.13 ꢀM, respectively). Moreover, the
methoxylated product 3 (R = 3,4-di-OCH3-C6H3) showed better antitumor activity being able to
inhibit the growth of 12 different cell lines, with a distinguished effect on the non-small cell lung
HOP-92 cell line at both the GI50 (0.05 ꢀM) and the TGI (9.77 ꢀM) levels, and a peculiar
efficiency against the leukaemia CCRF-CEM and SR cell lines (GI50 5.01 and 0.48 ꢀM,
respectively), Additionally, the same analog 3 exhibited an observable antitumor activity against
the leukaemia MOLT-4, non-small cell lung EKVX, CNS SNB-75 and breast BT-549 cell lines
(GI50 values 22.9, 30.9, 23.9 and 34.7 ꢀM, respectively). Replacement of the 6-aryl moiety with a
2-thienyl counterpart led to the moderately active compound 4 with variable growth inhibitory
effect on 12 different cell lines (GI50 range 10.0-83.2 ꢀM), and a particular effectiveness against
the non-small cell lung HOP-92 and renal UO-31 cell lines at both the GI50 (10.0 and 13.8 ꢀM,
respectively) and the TGI (40.7 and 27.5 ꢀM, respectively) levels. Moreover, a moderate
cytotoxic effect was shown by the same analog 4 against the renal UO-31 cell line, expressed by
LC50 value of 54.9 ꢀM. On the other hand, the structurally related analog 2-
oxotetrahydroquinoline-3-carbonitrile 5 revealed a dramatic reduction in the overall antitumor

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activity as expressed by the weak antitumor profile against against only 2 cell lines, namely; the
CNS SNB-75 and the ovarian OVCAR-4 cancer cell lines (GI50 39.8 and 69.2 ꢀM, respectively).
Shifting to the 2-aminopyridine-3-carbonitriles 6-8, compound 6 (R = C6H5) showed an
observable antitumor spectrum against 16 different cell lines, with a distinctive growth inhibitory
and cytostatic potentials against the leukaemia CCRF-CEM (GI50 0.08 and TGI 0.38 ꢀM) and
HL-60 (TB) (GI50 0.76 ꢀM), the non-small cell lung HOP-92 (GI50 0.01 and TGI 0.05 ꢀM), and
the CNS SNB-75 (GI50 18.6 and TGI 77.6 ꢀM) cell lines. Meanwhile, the methoxylated analog 7
(R = 3,4-di-OCH3-C6H3) revealed a wide range of growth inhibitory potential against 21 cancer
cell lines belonging to the 9 subpanels with a prominent effectiveness on all the tested leukaemia
subpanel tumor cell lines. In particular, compound 7 displayed pronounced growth inhibitory,
cytostatic and cytotoxic activities on the CCRF-CEM cell line (GI50, TGI and LC50 values 0.01,
3.98 and 9.77 ꢀM, respectively), in addition to a significant activity against the non-small cell
lung HOP-92 cell line (GI50 5.13 ꢀM). Furthermore, the 2-thienyl analog 8 showed mild to
moderate antitumor activity against 4 cell lines, with exceptional efficacy against the leukaemia
SR cell line at both the GI50 (5.37 ꢀM) and the TGI (67.6 ꢀM) levels. Finally, the bicyclic 2-
aminotetrahydroquinoline-3-carbonitrile derivative 9 was able to affect the growth of 14
different cell lines, with distinguished inhibitory action on the growth of the leukaemia HL-
60(TB) (GI50 0.42 ꢀM) and SR (GI50, TGI and LC50 values 3.39, 20.4 and 91.2 ꢀM, respectively)
cell lines.
Concerning the broad spectrum antitumor activity, the results revealed that the eight active
compounds (2-9) exhibited effective growth inhibition GI50 (MG-MID) values of 77.6, 67.6,
83.2, 91.2, 79.7, 60.3, 93.3 and 75.9 ꢀM, respectively. Moreover, six compounds namely; 2, 3, 4,
6, 7 and 9 were able to display a cytostatic activity with TGI (MG-MID) values of 85.1, 95.5,
95.5, 93.3, 79.4 and 97.7 ꢀM, respectively (Table 3). However, only two compounds namely; 6
and 7 showed weak cytotoxic efficacy with LC50 (MG-MID) values 95.5 and 93.3 ꢀM,
respectively).
2.2.3. Structure-activity correlation
A close examination of the structures of the biologically active compounds revealed that they
belong to four different series: monocyclic (2-oxo or 2-amino)pyridines (2-4 and 6-8), bicyclic

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tetrahydroquinolines (5, 9 and 10), tricyclic tetrahydropyrimido[4,5-b]quinolines (11, 12, 14 and
17) and a tetracyclic tetrahydropentaazacyclopenta[a]anthracene (18) (Schemes 1,2). In general,
the 2-oxo (or 2-amino)pyridines (2-4 and 6-8) exhibited significantly better antitumor spectrum
and potential being passed the primary anticancer assay, and carried over to the 5-dose screen.
According to the results recorded in table 3, it could be clearly recognized that better antitumor
activity was bounded to the 2-aminopyridine-3-carbonitrile scaffold (6-8). The nature of the aryl
moiety at C6 seems to remarkably affect the antitumor activity of these compounds as evidenced
by their GI50, TGI and LC50 values (Table 3). Among these, the 3,4-dimethoxyphenyl moiety (7;
R = 3,4-di-OCH3-C6H3) proved to be the most favourable substituent (GI50, TGI, and LC50 (MG-
MID) values 60.3, 79.4 and 93.3 ꢀM, respectively), when compared with its congeners 6 (R =
C6H5) and 8 (R = 2-thienyl). Analogously, the corresponding 6-(3,4-dimethoxyphenyl-2-
oxopyridine-3-carbonitrile analog 3 showed the highest activity among the the 2-oxopyridine-3-
carbonitrile series (2-4). With regard to the bicyclic tetrahydroquinolines (5, 9 and 10), the 2-
aminotetrahydroquinoline-3-carbonitrile 9 displayed considerable antitumor efficacy in the 5-
dose assay against 14 different cell lines, when compared with the 2-oxopyridine analog 5 which
showed limited activity against only 2 cell lines (Table 3). Introduction of a benzoylthioureido
counterpart at position 2, led to a less active compound 10 that couldn’t be promoted to the 5-
dose assay. However, it showed a broad spectrum of antitumor activity against 26 different
tumor cell lines (Table 2). Annulation of the bicyclic tetrahydroquinoline 9 into variously
substituted tricyclic tetrahydropyrimido[4,5-b]quinolines furnished 4 compounds namely; 11, 12,
14 and 17. Among these, the 1H-pyrimido[4,5-b]quinoline-2-thione 11 and the ethyl thioether 12
exerted a noticeable broad spectrum of growth inhibitory activity on a variety of cell lines
belonging to different tumor subpanels, with obvious effect on the leukaemia subpanel tumor cell
lines. The N-phenylpyrimido[4,5-b]quinoline-2-thione 14 and iminopyrimido[4,5-b]quinoline 17
possessed moderate to weak antitumor activity, with special potency against the leukemia cell
lines. However, conversion of the imino derivative 17 into the tetracyclic pentaazacyclopenta[a]
anthracene 18 resulted in a reduction in the overall antitumor potency and spectrum.
2.3. Antioxidant Activity
DPPH radical scavenging assay
Compounds 3, 6, 7, 9, 10, 11 and 12 which exhibited prominent antitumor activity, were further
evaluated for their antioxidant potential through scavenging of the 2,2-diphenyl-1-picrylhydrazyl

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(DPPH) radical, according to a modified procedure from that described by Blois [51]. The
principle of this assay is based on the fact that the odd electron in DPPH free radical gives a
strong absorption maximum in the visible region at λmax 517 nm (violet in color). Upon
reduction in the presence of an antioxidant, the violet color switches to yellow when such odd
electron of DPPH radical conjugates with a hydrogen to form the reduced DPPH-H (2,2-
diphenyl-1-picrylhydrazine). This color change is stoichiometric, where the degree of
discoloration is proportional to the scavenging potential of the tested compound i.e. low
absorbance refers to remarkable free radical scavenging potential. The antioxidant potential of
the tested compounds was calculated as the percentage DPPH scavenging effect in relation to
control and butylated hydroxyl toluene (BHT) was utilized as a reference standard antioxidant.
The results recorded in table 4 revealed that all the tested seven compounds showed variable
degree of DPPH scavenging activity. At the 10-3 M concentration level, the substituted 6-(3,4-
dimethoxyphenyl)pyridine-3-carbonitriles 3 and 7 (scavenging activity 31 and 33 %,
respectively) were nearly equiactive with BHT; the standard antioxidant utilized in this assay
(scavenging activity 36 %), whereas, the substituted 6-phenyl-2-aminopyridine-3-carbonitrile
analog 6 displayed an appreciable activity (scavenging activity 27 %). Nevertheless, the
structurally related 2-aminotetrahydroquinoline-3-carbonitrile 9 displayed a relatively weak
antioxidant activity (scavenging activity 16 %), when compared with compounds 3, 6 and 7. On
the other hand, the sulphur-containing benzoylthioureidoquinoline 10, 1H-pyrimido[4,5-
b]quinoline-2-thione 11 and the derived ethyl thioether 12 exhibited considerable activity
(scavenging activity 23, 26 and 28 %, respectively). In a similar pattern, the same order of DPPH
radical scavenging potential was expressed at the 10-4 M concentration level, with a scavenging
activity range 9-19 %, when compared with BHT (scavenging activity 20 %).
3. Conclusion
The main aim of this investigation was to comparatively study the effect of utilizing both the
conventional heating and MW irradiation techniques in the synthesis of some novel

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pyrrolylpyridines and some derived bi-, tri- and tetracyclic ring systems, to be biologically
evaluated as anticancer and antioxidant agents. The obtained NCI’s in vitro one-dose antitumor
data revealed that the analogs 10-12, 14, 17 and 18 showed various growth inhibitory (GI %)
profiles against some of the tested subpanel tumor cell lines. Meanwhile, the analogs 2-9 have
successfully passed the primary one-dose screening, and promoted to the in vitro full panel (five-
dose) 60-cell line assay. The best antitumor activity was bounded to the 2-aminopyridine-3-
carbonitrile scaffold (6-8) especially 7 (R = 3,4-di-OCH3-C6H3), whereas the bicyclic
tetrahydroquinolines (5, 9 and 10) displayed considerable antitumor efficacy. Further annulation
of the bicyclic tetrahydroquinoline 9 into variously substituted tricyclic tetrahydropyrimido[4,5-
b]quinolines (11, 12, 14 and 17) and the tetracyclic pentaazacyclopenta[a]anthracene 18 resulted
in a reduction in the overall antitumor potency and spectrum, being not able to pass to the 5-dose
assay. On the other hand, compounds 3, 6, 7, 9, 10, 11 and 12 which exhibited prominent
antitumor activity, were further evaluated for their antioxidant potential using the DPPH radical
scavenging assay. At the 10-3
M
concentration level, the substituted 6-(3,4-
dimethoxyphenyl)pyridine-3-carbonitriles 3 and 7 showed scavenging activity 31 and 33 %,
respectively, which were nearly equiactive with BHT; the standard antioxidant utilized in this
assay (scavenging activity 36 %). Finally, the broad spectrum antitumor activity and special
efficacy against leukaemia together with the reliable antioxidant potential displayed by
compounds 3 and 7 will be of interest for future derivatization in the hope of finding more active
and selective lead structures for dual antitumor and antioxidant activities.
4. Experimental
4.1. Chemistry
Microwave-assisted syntheses were carried out in a (CEM Discover, 300W) microwave
synthesizer using a pressure tube (10 mL) sealed with a plastic cap. Melting points were
determined on a Gallenkamp melting point apparatus and are uncorrected. The infrared (IR)
spectra were recorded on Shimadzu FT-IR 8400S infrared spectrophotometer using the KBr
1
pellet technique. H and 13C-NMR spectra were recorded on a Bruker DPX-400 FT NMR
spectrometer using tetramethylsilane as the internal standard and a mixture of CDCl3 and

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DMSO-d6 as a solvent (Chemical shifts in δ, ppm). Splitting patterns were designated as follows:
s: singlet; d: doublet; m: multiplet; q: quartet. Elemental analyses were performed on a 2400
Perkin Elmer Series 2 analyzer and the found values were within ±0.4% of the theoretical values.
Follow up of the reactions and checking the homogeneity of the compounds were made by TLC
on silica gel-protected aluminum sheets (Type 60 F254, Merck) and the spots were detected by
exposure to UV-lamp at λ 254.
4.1.1. General procedure for the synthesis of 4-(1-methyl-1H-pyrrol-2-yl)-2-oxo-6-
substituted-1,2-dihydropyridine-3-carbonitriles (2-4) and 8-methyl-4-(1-methyl-1H-pyrrol-
2-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinoline-3-carbonitrile (5)
Conventional heating: a one-pot mixture of 1-methyl-1H-pyrrole-2-carbaldehyde 1 (0.55 g, 5
mmol), the appropriate ketone (5 mmol), ethyl cyanoacetate (0.57 g, 5 mmol) and ammonium
acetate (3.1 g, 40 mmol) in absolute ethanol (20 mL), was refluxed for 5-6 h. The reaction
mixture was allowed to cool, and the formed solid product was filtered, washed with water, dried
and recrystallized from ethanol.
Microwave irradiation: a thoroughly mixed solvent-free mixture of the aldehyde 1 (0.55 g, 5
mmol), the appropriate ketone (5 mmol), ethyl cyanoacetate (0.57 g, 5 mmol) and ammonium
O
acetate (3.1 g, 40 mmol), was subjected to microwave irradiation at 100 C for 15 min. After
being cooled to room temperature, the residue was treated with water, filtered, dried and
recrystallized from ethanol. IR (cm-1): 3380-3220 (NH), 2220- 2210 (CN), 1685-1675 (C=O).
Yields and melting points are recorded in table 1.
4.1.1.1. 4-(1-Methyl-1H-pyrrol-2-yl)-2-oxo-6-phenyl-1,2-dihydropyridine-3-carbonitrile (2).
1H-NMR (δ, ppm): 3.46 (s, 3H, N-CH3), 6.38 (s, 1H, pyridine C5-H), 6.92-7.36 (m, 8H, 3
pyrrole-H and 5 Ar-H), 8.15 (s, 1H, NH). 13C-NMR (δ, ppm): 45.6 (CH3), 118.3 (CN), 89.2,
102.2, 110.8, 116.7, 121.4, 127.5, 128.8, 130.4, 132.8, 133.3, 144.3, 162.9, 166.1 (Ar C), 169.5
(CO). Anal. Calcd. for C17H13N3O (275.3): C, 74.17; H, 4.76; N, 15.26. Found: C, 73.94; H,
4.93; N, 15.11.
4.1.1.2. 6-(3,4-Dimethoxyphenyl)-4-(1-methyl-1H-pyrrol-2-yl)-2-oxo-1,2-dihydropyridine-3-
1
carbonitrile (3). H-NMR (δ, ppm): 3.49 (s, 3H, N-CH3), 3.85 (s, 3H, OCH3), 3.90 (s, 3H,
OCH3), 6.47 (s, 1H, pyridine C5-H), 6.76-7.41 (m, 6H, 3 pyrrole-H and 3 Ar-H), 8.18 (s, 1H,
NH). 13C-NMR (δ, ppm): 46.7 (CH3), 55.3 (OCH3), 56.9 (OCH3), 117.9 (CN), 88.9, 102.3,

ACCEPTED MANUSCRIPT
13
111.5, 116.6, 120.1, 125.8, 129.5, 130.7, 132.8, 133.9, 145.8, 162.9, 164.4 (Ar C), 169.7 (CO).
Anal. Calcd. for C19H17N3O3 (335.36): C, 68.05; H, 5.11; N, 12.53. Found: C, 68.37; H, 5.02; N,
12.44.
4.1.1.3. 4-(1-Methyl-1H-pyrrol-2-yl)-2-oxo-6-(2-thienyl)-1,2-dihydropyridine-3-carbonitrile
1
(4). H-NMR (δ, ppm): 3.47 (s, 3H, N-CH3), 6.40 (s, 1H, pyridine C5-H), 6.81-7.24 (m, 6H, 3
pyrrole-H and 3 thiophene-H), 8.17 (s, 1H, NH). 13C-NMR (δ, ppm): 48.1 (CH3), 119.0 (CN),
88.9, 101.9, 110.8, 118.3, 122.5, 127.4, 131.2, 142.0, 146.3, 162.3, 166.4 (Ar C), 170.1 (CO).
Anal. Calcd. for C15H11N3OS (281.33): C, 64.04; H, 3.94; N, 14.94; S, 11.4. Found: C, 63.87; H,
4.05; N, 14.82; S, 11.21.
4.1.1.4.
8-Methyl-4-(1-methyl-1H-pyrrol-2-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinoline-3-
carbonitrile (5) 1H-NMR (δ, ppm): 1.38 (s, 3H, CH3), 1.64-2.40 (m, 6H, C5,6,7-H), 2.83 (m, 1H,
13
C8-H), 3.39 (s, 3H, N-CH3), 6.89-7.21 (m, 3H, 3 pyrrole-H), 8.26 (s, 1H, NH). C-NMR (δ,
ppm): 16.1 (CH3), 27.7, 27.9, 34.4, 34.9 (cyclohexyl C), 45.4 (CH3), 118.4 (CN), 95.7, 102.2,
110.8, 116.5, 121.4, 126.3, 132.7, 169.4 (Ar C), 169.9 (CO). Anal. Calcd. for C16H17N3O
(267.33): C, 71.89; H, 6.41; N, 15.72. Found: C, 72.06; H, 6.33; N, 15.48.
4.1.2. General procedure for the synthesis of 2-amino-4-(1-methyl-1H-pyrrol-2-yl)-6-
substituted nicotinonitrile (6-8) and 2-amino-8-methyl-4-(1-methyl-1H-pyrrol-2-yl)-5,6,7,8-
tetrahydroquinoline-3-carbonitrile (9).
Conventional heating: a one-put mixture of the aldehyde 1 (0.55 g, 5 mmol), the appropriate
ketone (5 mmol), malononitrile (0.33 g, 5 mmol) and ammonium acetate (3.1 g, 40 mmol) in
absolute ethanol (20 mL), was heated under reflux for 4-6 h. After cooling, the precipitated
product was filtered, washed with water, dried and recrystallized from ethanol.
Microwave irradiation: a thoroughly mixed solvent-free mixture of the aldehyde 1 (0.55 g, 5
mmol), the appropriate ketone (5 mmol), malononitrile (0.33 g, 5 mmol) and ammonium acetate
O
(3.1 g, 40 mmol), was subjected to microwave irradiation at 100 C for 10 min. After being
cooled to room temperature, the residue was treated with water, filtered, dried and recrystallized
from ethanol. IR (cm-1): 3390-3240 (NH), 2225-2210 (CN). Yields and melting points are
recorded in table 1.
4.1.2.1. 2-Amino-4-(1-methyl-1H-pyrrol-2-yl)-6-phenylnicotinonitrile (6). 1H-NMR (δ, ppm):
3.46 (s, 3H, N-CH3), 5.09 (s, 2H, NH2), 6.41 (s, 1H, pyridine C5-H), 6.89-7.21 (m, 8H, 3 pyrrole-

ACCEPTED MANUSCRIPT
14
H and 5 Ar-H). 13C-NMR (δ, ppm): 48.3 (CH3), 119.9 (CN), 89.1, 102.2, 110.8, 118.3, 121.4,
127.5, 128.8, 130.4, 132.8, 133.3, 146.3, 161.9, 167.2 (Ar C). Anal. Calcd. for C17H14N4
(274.32): C, 74.43; H, 5.14; N, 20.42. Found: C, 74.80; H, 5.09; N, 20.12.
4.1.2.2. 6-(3,4-Dimethoxyphenyl)-2-amino-4-(1-methyl-1H-pyrrol-2-yl)-nicotinonitrile (7).
1H-NMR (δ, ppm): 3.45 (s, 3H, N-CH3), 3.89 (s, 6H, 2 OCH3), 5.06 (s, 2H, NH2), 6.44 (s, 1H,
pyridine C5-H), 6.81-7.26 (m, 6H, 3 pyrrole-H and 3 Ar-H). 13C-NMR (δ, ppm): 47.9 (CH3), 54.2
(OCH3), 56.6 (OCH3), 118.3 (CN), 89.7, 101.9, 111.2, 116.3, 120.9, 126.6, 129.1, 131.2, 133.6,
134.1, 145.6, 162.5, 167.9 (Ar C). Anal. Calcd. for C19H18N4O2 (334.37): C, 68.25; H, 5.43; N,
16.76. Found: C, 68.53; H, 5.23; N, 16.93.
1
4.1.2.3. 2-Amino-4-(1-methyl-1H-pyrrol-2-yl)-6-(2-thienyl)nicotinonitrile (8). H-NMR (δ,
ppm): 3.46 (s, 3H, N-CH3), 5.07 (s, 2H, NH2), 6.42 (s, 1H, pyridine C5-H), 6.79-7.20 (m, 6H, 3
13
pyrrole-H and 3 thiophene-H). C-NMR (δ, ppm): 48.1 (CH3), 118.6 (CN), 89.1, 101.2, 110.2,
118.3, 121.9, 126.5, 130.4, 141.8, 146.3, 161.9, 167.2 (Ar C). Anal. Calcd. for C15H12N4S
(280.35): C, 64.26; H, 4.31; N, 19.98; S, 11.44. Found: C, 64.41; H, 4.42; N, 20.28; S, 11.63.
4.1.2.4.
2-Amino-8-methyl-4-(1-methyl-1H-pyrrol-2-yl)-5,6,7,8-tetrahydroquinoline-3-
carbonitrile (9). 1H-NMR (δ, ppm): 1.36 (s, 3H, CH3), 1.62-2.37 (m, 6H, C5,6,7-H), 2.86 (m, 1H,
C8-H), 3.36 (s, 3H, N-CH3), 5.06 (s, 2H, NH2), 6.89-7.21 (m, 3H, 3 pyrrole-H). 13C-NMR (δ,
ppm): 21.1 (CH3), 22.1, 27.6, 31.2, 38.4 (C5,6,7,8), 46.2 (CH3), 118.7 (CN), 88.1, 102.2, 110.8,
117.3, 121.4, 145.7, 163.8, 164.2, (Ar C). Anal. Calcd. for C16H18N4 (266.34): C, 72.15; H, 6.81;
N, 21.04. Found: C, 72.33; H, 7.06; N, 21.16.
4.1.3.
N-((3-cyano-8-methyl-4-(1-methyl-1H-pyrrol-2-yl)-5,6,7,8-tetrahydroquinolin-2-
yl)carbamothioyl)benzamide (10).
Conventional heating: to a mixture of the 2-amino-8-methyl-4-(1-methyl-1H-pyrrol-2-yl)-
5,6,7,8-tetrahydroquinoline-3-carbonitrile 9 (1.33 g, 5 mmol) and K2CO3 (0.9 g, 6.5 mmol) in
dry acetone (20 mL), a solution of benzoyl isothiocyanate (0.82 g, 5 mmol) in dry acetone (5
mL) was added. The resultant reaction mixture was heated under reflux for 2 h, then left for an
overnight at room temperature, concentrated to half its volume and allowed to cool in the
refrigerator for 4 h. The separated solid product was filtered, washed with diethyl ether and
crystallized from ethanol.

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15
Microwave irradiation: a mixture of the tetrahydroquinoline 9 (1.33 g, 5 mmol), K2CO3 (0.9 g,
6.5 mmol), benzoyl isothiocyanate (0.82 g, 5 mmol) in dry acetone (10 mL), was subjected to
O
microwave irradiation at 100 C for 10 min. After being cooled to room temperature, the
reaction mixture was worked up as described under the conventional heating procedure. Yields
and melting point are recorded in table 1. IR (cm-1): 3480-3330 (NH), 2220 (CN), 1710 (C=O),
1170 (C=S). 1H-NMR (δ, ppm): 1.38 (s, 3H, CH3), 1.64-2.64 (m, 6H, C5,6,7-H), 3.05 (m, 1H, C8-
H), 3.40 (s, 3H, N-CH3), 5.05 (s, 1H, NH), 7.09-7.73 (m, 8H, 3 pyrrole-H and Ar-H), 7.99 (s,
1H, NH). 13C-NMR (δ, ppm): 21.4 (CH3), 25.4, 31.5, 32.2, 39.1 (C5,6,7,8), 56.1 (CH3), 91.9,
104.5, 110.3, 121.7, 126.2, 128.8, 130.4, 131.8, 133.5, 156.7, 161.5, 163.7 (Ar C), 118.0 (CN),
180 (CS), 170.1 (CO). Anal. Calcd. for C24H23N5OS (429.54): C, 67.11; H, 5.40; N, 16.30; S,
7.47. Found: C, 67.38; H, 5.24; N, 16.42; S, 7.63.
4.1.4. 4-Amino-9-methyl-5-(1-methyl-1H-pyrrol-2-yl)-6,7,8,9-tetrahydro-1H-pyrimido[4,5-
b]quinoline-2-thione (11).
Conventional method: the key intermediate 9 (1.33 g, 5 mmol) and thiourea (1.9 g, 25 mmol)
were fused at 260-300 ºC using a sand bath for 1 h. The reaction mixture was allowed to attain
room temperature, the crude solid product was rubbed with cold ethanol, filtered and
recrystallized from DMF containing few drops of water.
Microwave method: a mixture of 9 (1.33 g, 5 mmol) and thiourea (1.9 g, 25 mmol), was
subjected to microwave irradiation at 250 OC for 10 min. After being cooled to room
temperature, the reaction mixture was worked up as described under the conventional heating
procedure. Yields and melting point are recorded in table 1. IR (cm-1): 3345-3280 (NH), 1180
1
(C=S). H-NMR (δ, ppm): 1.39 (s, 3H, CH3), 1.63-2.44 (m, 6H, C6,7,8-H), 3.18 (m, 1H, C9-H),
3.56 (s, 3H, N-CH3), 5.06 (s, 2H, NH2), 6.88-7.29 (m, 4H, 3 pyrrole-H and N-H). 13C-NMR (δ,
ppm): 21.1 (CH3), 26.2, 32.5, 33.1, 38.6 (C6,7,8,9), 52.0 (CH3), 109.7, 114.4, 124.6, 127.8, 130.3,
149.6, 156.9, 162.5, 163.4, 164.3 (Ar C), 182.8 (CS). Anal. Calcd. for C17H19N5S (325.43): C,
62.74; H, 5.88; N, 21.52; S, 9.85. Found: C, 62.65; H, 5.94; N, 21.39; S, 10.07.

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4.1.5.
2-Alkylthio-4-amino-9-methyl-5-(1-methyl-1H-pyrrol-2-yl)-6,7,8,9-tetrahydro-1H-
pyrimido[4,5-b]quinolines (12 and 13).
Conventional heating: to a stirred solution of the thione 11 (0.98 g, 3 mmol) in a mixture of 1N
NaOH (10 mL) and ethanol (5 mL), was added the appropriate alkyl halide (4.5 mmol). The
reaction mixture was stirred at room temperature for 6 h, and the precipitated product was
filtered, washed with aqueous ethanol, dried and recrystallized from ethanol.
Microwave irradiation: a solution of the thioureido derivative 10 (1.23 g, 3 mmol) and the
appropriate alkyl halide (4.5 mmol) in a mixture of 1N sodium hydroxide (3 mL) and EtOH (7
O
mL), was subjected to microwave irradiation at 180 C for 15 min. After being cooled to room
temperature, the reaction mixture was worked up as described under the conventional heating
procedure. Yield and melting point are recorded in table 1. IR (cm-1): 3440-3150 (NH).
4.1.5.1. 4-Amino-2-ethylthio-9-methyl-5-(1-methyl-1H-pyrrol-2-yl)-6,7,8,9-tetrahydro-1H-
1
pyrimido[4,5-b]quinoline (12). H-NMR (δ, ppm): 1.25 (t, J = 9 Hz, 3H, ethyl-CH3), 1.39 (s,
3H, CH3), 1.41-2.23 (m, 6H, C6,7,8-H), 3.21 (m, 1H, C9-H), 3.52 (q, J = 9 Hz, 2H, ethyl-CH2),
13
3.51 (s, 3H, N-CH3), 5.28 (s, 2H, NH2), 6.88-7.21 (m, 3H, pyrrole-H). C-NMR (δ, ppm): 15.8
(CH3), 22.1 (CH3), 29.4 (CH2), 25.8, 30.9, 32.7, 37.9 (C6,7,8,9), 55.3 (CH3), 102.3, 113.4, 128.9,
130.4, 135.4, 148.7, 151.4 158.0, 162.5, 164.2, 167.6 (Ar C). Anal. Calcd. for C19H23N5S
(353.48): C, 64.56; H, 6.56; N, 19.81; S, 9.07. Found: C, 64.83; H, 6.37; N, 19.64; S, 9.33.
4.1.5.2. 4-Amino-2-benzylthio-9-methyl-5-(1-methyl-1H-pyrrol-2-yl)-6,7,8,9-tetrahydro-1H-
1
pyrimido[4,5-b]quinoline (13). H-NMR (δ, ppm): 1.36 (s, 3H, CH3), 1.56-2.31 (m, 6H, C6,7,8
-
H), 3.18 (m, 1H, C9-H), 3.46 (s, 3H, N-CH3), 4.25 (s, 2H, benzyl CH2), 5.32 (s, 2H, NH2), 6.82-
7.58 (m, 8H, 3 pyrrole-H and Ar-H). 13C NMR (δ-ppm): 15.4 (CH3), 30.8 (N-CH3), 44.9 (benzyl-
CH2), 27.6, 28.2, 32.2, 34.9 (C6,7,8,9), 95.8, 104.0, 111.2, 119.7, 121.2, 122.6, 124.1, 125.3, 126.8,
128.8, 129.3, 132.7, 161.6 (Ar C). Anal. Calcd. for C24H25N5S (415.55): C, 69.37; H, 6.06; N,
16.85; S, 7.72. Found: C, 69.51; H, 6.32; N, 16.94; S, 7.91.
4.1.6.
4-Imino-9-methyl-3-phenyl-5-(1-methyl-1H-pyrrol-2-yl)-3,4,6,7,8,9-hexahydro-1H-
pyrimido [4,5-b]quinoline-2-thione (14).
Conventional method: a mixture of 9 (1.33 g, 5 mmol), phenyl isothiocyanate (1 g, 7.5 mmol)
in pyridine (10 mL) was refluxed for 3 h. After cooling, the separated solid product was filtered
off, washed thoroughly with water, dried and recrystallized from acetic acid.

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17
Microwave method: a mixture of 9 (1.33 g, 5 mmol), phenyl isothiocyanate (1 g, 7.5 mol) and
pyridine (5 mL) was subjected to microwave irradiation at 180 OC for 15 min. After being cooled
to room temperature, the reaction mixture was worked up as described under the conventional
heating procedure. Yields and melting point are recorded in table 1. IR (cm-1): 3330-3160 (NH),
1635 (C=N), 1210 (C=S). 1H-NMR (δ, ppm): 1.39 (s, 3H, CH3), 1.68-2.39 (m, 6H, C6,7,8-H), 3.12
(m, 1H, C9-H), 3.45 (s, 3H, N-CH3), 4.98 (s, 1H, NH), 6.81-7.52 (m, 8H, 3 pyrrole-H and Ar-H),
13
8.05 (s, 1H, NH). C-NMR (δ-ppm): 21.6 (CH3), 26.4, 32.6, 34.5, 39.7 (C6,7,8,9), 51.2 (CH3),
91.3, 109.8, 114.4, 125.2, 127.6, 128.6, 130.5, 139.7, 148.8, 157.5 (Ar C), 165.2 (C=NH), 179.4
(CS). Anal. Calcd. for C23H23N5S (401.53): C, 68.8; H, 5.77; N, 17.44; S, 7.99. Found: C, 68.54;
H, 5.37; N, 17.69; S, 8.16.
4.1.7.
2,9-Dimethyl-5-(1-methyl-1H-pyrrol-2-yl)-6,7,8,9-tetrahydro-3H-pyrimido[4,5-b]-
quinolin-4-one (15). A mixture of 9 (1.33 g, 5 mmol), acetic anhydride (10 mL) and conc.
H2SO4 (1 mL) was heated in a boiling water bath for 10 min. The reaction mixture was cooled,
poured carefully onto ice-cold water, treated with 20% NaOH solution till alkaline. The
precipitated crude solid product was filtered, washed with water, dried and recrystallized from
ethanol. Yield and melting point are recorded in table 1. IR (cm-1): 3430-3220 (NH), 1715
(C=O).1H-NMR (δ, ppm): 1.38 (s, 3H, CH3), 1.46-2.2 (m, 6H, C6,7,8-H), 2.56 (s, 3H, C2-CH3),
3.14 (m, 1H, C9-H), 3.88 (s, 3H, N-CH3), 6.87-7.51 (m, 3H, 3 pyrrole-H), 8.03 (s, 1H, NH).
13CNMR (δ, ppm): 18.9 (CH3), 21.4 (CH3), 22.1, 28.4, 31.2, 38.7 (C6,7,8,9), 55.4 (CH3), 113.1,
119.2, 131.5, 134.2, 139.5, 152.6, 159.0, 161.4, 162.5, 169.3 (Ar C), 174.5 (CO). Anal. Calcd.
for C18H20N4O (308.38): C, 70.11; H, 6.54; N, 18.17. Found: C, 69.93; H, 6.31; N, 18.32.
4.1.8.
2-Ethoxymethylidineamino-3-cyano-8-methyl-4-(1-methyl-1H-pyrrol-2-yl)-5,6,7,8-
tetrahydroquinoline (16).
Conventional method: a mixture of the tetrahydroquinoline 9 (1.33 g, 5 mmol),
triethylorthoformate (5 mL) and acetic anhydride (3 mL), was heated for 10 h. The reaction
mixture was allowed to cool then diluted with cold water. The precipitated solid was collected by
filtration, washed with water and recrystallized from ethanol/benzene mixture 3:1.
Microwave method: a mixture of the tetrahydroquinoline 9 (1.33 g, 5 mmol),
triethylorthoformate (5 mL) and acetic anhydride (3 mL), was subjected to microwave irradiation

ACCEPTED MANUSCRIPT
18
at 180 OC for 20 min. After being cooled to room temperature, the reaction mixture was worked
up as described in the conventional procedure. Yields and melting point are recorded in table 1.
1
IR (cm-1): 2223 (CN), 1620 (C=N). H-NMR (δ, ppm): 1.25 (t, J = 8.6, 3H, ethyl-CH3), 1.36 (s,
3H, CH3), 1.69-2.31 (m, 6H, C5,6,7-H), 3.09 (m, 1H, C8-H), 3.69 (q, J = 8.6, 2H, ethyl-CH2), 3.81
13
(s, 3H, N-CH3), 6.87-7.29 (m, 3H, pyrrole-H), 8.11 (s, 1H, N=CH). C-NMR (δ, ppm): 15.5
(CH3), 21.2 (CH3), 23.2, 28.4, 34.5, 38.6 (cyclohexyl C), 54.1 (CH3), 58.3 (CH2), 117.7 (CN),
103.1, 115.8, 127.2, 130.8, 134.2, 155.5, 162.3, 169.1, 171.8 (Ar C), 165.1 (N=C). Anal. Calcd.
for C19H22N4O (322.4): C, 70.78; H, 6.88; N, 17.38. Found: C, 70.92; H, 6.57; N, 17.64.
4.1.9.
3-Amino-4-imino-9-methyl-5-(1-methyl-1H-pyrrol-2-yl)-6,7,8,9-tetrahydro-4H-
pyrimido[4,5-b]quinoline (17).
Conventional method: a mixture of the intermediate tetrahydroquinoline 16 (1.6 g, 5 mmol) and
hydrazine hydrate 99% (1 g, 20 mmol) in dry benzene (30 mL), was heated under reflux for 4 h.
On cooling, the solid product thus obtained was filtered, dried and recrystallized from toluene.
Microwave method: a solvent-free mixture of the intermediate tetrahydroquinoline 16 (1.6 g, 5
mmol) and hydrazine hydrate 99% (1 g, 20 mmol) was subjected to microwave irradiation at 150
OC for 10 min. After being cooled to room temperature, the reaction mixture was worked up as
described under the conventional heating procedure. Yields and melting point are recorded in
table 1. IR (cm-1): 3350-3235 (NH), 1640 (C=N). 1H-NMR (δ, ppm): 1.38 (s, 3H, CH3), 1.58-
2.31(m, 6H, C6,7,8-H), 2.85 (m, 1H, C9-H), 3.81 (s, 3H, N-CH3), 5.1 (s, 2H, NH2), 6.84-7.31 (m,
4H, pyrrole-H and C2-H), 7.91 (s, 1H, NH). 13C-NMR (δ, ppm): 21.1 (CH3), 22.4, 27.4, 32.2,
39.4 (C6,7,8,9), 56.6 (CH3), 113.8, 119.3,2, 127.8, 130.4, 134.2, 147.1, 159.4, 161.7, 162.1, 164.0,
167.8 (Ar C). Anal. Calcd. for C17H20N6 (308.38): C, 66.21; H, 6.54; N, 27.25. Found: C, 66.13;
H, 6.87; N, 27.61.
4.1.10.
7-Methyl-11-(1-methyl-1H-pyrrol-2-yl)-7,8,9,10-tetrahydro-1,3,3a,5,6-
pentaazacyclopenta[a]anthracene (18).
Conventional heating: a solution of the pyrimido[4,5-b]quinoline 17 (1.54 g, 5 mmol) in formic
acid (10 mL) was heated under reflux for 6 h. The reaction mixture was cooled to room
temperature, diluted with cold water and the precipitated solid was collected by filtration,
washed with water, dried and recrystallized from benzene. IR (cm-1): 1630 (C=N).

ACCEPTED MANUSCRIPT
19
Microwave irradiation: a mixture of 17 (1.54 g, 5 mmol) and formic acid (10 mL) was
subjected to microwave irradiation at 150 OC for 20 min. After being cooled to room
temperature, the reaction mixture was worked up as described under the conventional heating
procedure. Yields and melting point are recorded in table 1. IR (cm-1): 1640 (C=N). 1H-NMR (δ,
ppm): 1.38 (s, 3H, CH3), 1.54-2.41 (m, 6H, C8,9,10-H), 3.1 (m, 1H, C7-H), 3.86 (s, 3H, N-CH3),
13
6.91-7.41 (m, 3H, pyrrole-H), 8.69 (s, 1H, C2-H), 9.18 (s, 1H, C4-H). C-NMR (δ, ppm): 21.8
(CH3), 22.4, 27.9, 32.9, 40.2 (C7,8,9,10), 55.2 (CH3), 115.6, 122.3, 126.5, 131.7, 136.1, 145.3,
147.1, 155.3, 158.6, 162.5, 163.8 (Ar C). Anal. Calcd. for C18H18N6 (318.38): C, 67.9; H, 5.7; N,
26.4. Found: C, 68.08; H, 5.52; N, 26.51.
4.1.11.
2,7-Dimethyl-11-(1-methyl-1H-pyrrol-2-yl)-7,8,9,10-tetrahydro-1,3,3a,5,6-
pentaazacyclopenta[a]anthracene (19).
Conventional heating: a mixture of 17 (1.54 g, 5 mmol) and acetic anhydride (10 mL) was
heated under reflux for 8 h. The reaction mixture was allowed to attain room temperature, diluted
with cold water, and the precipitated solid was filtered, washed with water, dried and
recrystallized from acetic acid.
Microwave irradiation: a mixture of 17 (1.54 g, 5 mmol) and acetic anhydride (10 mL) was
subjected to microwave irradiation at 200 OC for 20 min. After being cooled to room
temperature, the reaction mixture was worked up as described under the conventional heating
1
procedure. Yields and melting point are recorded in table 1. IR (cm-1): 1626 (C=N). H-NMR (δ,
ppm): 1.41 (s, 3H, CH3), 1.49-2.47 (m, 6H, C8,9,10-H), 2.55 (s, 3H, C2-CH3), 3.07 (m, 1H, C7-H),
3.85 (s, 3H, N-CH3), 6.91-7.41 (m, 3H, pyrrole-H), 9.01 (s, 1H, C4-H). 13C-NMR (δ, ppm): 14.8
(CH3), 20.9 (CH3), 21.8, 27.3, 32.2, 38.9 (C7,8,9,10), 56.8 (CH3), 113.2, 122.4, 127.8, 132.4, 136.5,
147.6, 148.9, 155.4, 158.1, 162.2, 163.9 (Ar C). Anal. Calcd. for C19H20N6 (332.4): C, 68.65; H,
6.06; N, 25.28. Found: C, 68.34; H, 6.22; N, 25.39.
4.2. In vitro antitumor screening
Out of the newly synthesized compounds, fourteen derivatives namely: 2-12, 14, 17 and 18; were
selected by the National Cancer Institute (NCI) in-vitro disease-oriented human cells screening
panel assay [48-50] to be evaluated for their in-vitro antitumor activity. Primary in vitro one dose
anticancer assay was performed using the full NCI 60 cell panel in accordance with the current

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20
protocol of the Drug Evaluation Branch, NCI, Bethesda, Maryland, USA. These cell lines were
incubated with one concentration (10 ꢀM) for each tested compound. A 48h continuous drug
exposure protocol was used, and a sulphorhodamine B (SRB) protein assay was employed to
estimate cell viability or growth. Data are reported as a mean graph of the percent growth of
treated cells, and presented as percentage growth inhibition (GI %) caused by the test compounds
(Table 2).
Compound 2-9, passed this primary anticancer assay and consequently was carried over to the 5-
dose screen against a panel of about 60 different tumor cell lines of nine tumor subpanels,
including leukemia, non-small cell lung, colon, CNS, melanoma, ovarian, renal, prostate and
breast cancer cell lines. These cell lines were incubated with five concentrations (0.01-100 ꢀM)
for the tested compound and were used to create log concentration % growth inhibition curves.
Three response parameters: GI50 (growth inhibitory activity), TGI (cytostatic activity), and LC50
(cytotoxic activity) were calculated for each cell line, using 5-flourouracil (5-FU) as a positive
control. Subpanel and full panel mean-graph midpoint values (MG-MID) for certain agents are
the average of individual real and default GI50, TGI, or LC50 values of all cell lines in the
subpanel or the full panel, respectively. (Table 3).
4.3. DPPH radical scavenging antioxidant assay
Compounds 3, 6, 7, 9, 10, 11 and 12 which exhibited prominent antitumor activity, were tested
for their ability to show antioxidant effect through scavenging of the 2,2-diphenyl-1-
picrylhydrazyl (DPPH) radical, according to a modified procedure from that described by Blois
[51]. Two concentrations of the tested compounds (10-3 and 10-4 M) were mixed with a
methanolic solution of DPPH (0.1 mL of 1 mM) at room temperature, so that the total volume of
the reaction mixture is 3 mL. The mixture was shaken vigorously and allowed to stand in the
dark for 30 min at room temperature. Thereafter, the absorbance (A) of the obtained solution was
measured spectrophotometrically in the visible region at λmax 517 nm. The same procedure was
performed for a control which is DPPH radical solution in methanol alone. BHT was utilized as a
reference standard antioxidant in this experiment. Each experiment was carried out in triplicate.

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The capability of the tested compounds to scavenge DPPH radical was calculated according
to the following equation:
DPPH scavenging effect (%) = (AC – AS) / AC × 100
Where AC is the absorbance of control (DPPH radical solution in methanol), and AS represents the
value of the absorbance of the sample (solution of DPPH radical and tested compound in methanol).
The obtained data are presented in table 4.
Acknowledgement
This paper was funded by the Deanship of Scientific Research (DSR), King Abdulaziz
University, Jeddah, Kingdom Saudi Arabia, under grant number (166-224-D1435). The authors,
therefore, acknowledge with thanks DSR technical and financial support.
Sincere gratitude is conveyed to the staff members of the Department of Health and Human
Services, National Cancer Institute (NCI), Bethesda, Maryland, U.S.A. for carrying out the
anticancer screening of the newly synthesized compounds.
References
[1]
[2]
[3]
I.J. Fidler, Nat. Rev. Cancer 3 (2003) 453-458.
B.A. Gilchrest, M.S. Eller, Drugs Future 34 (2009) 205-216.
G. Garrido, D. Gonzalez, C. Delporte, N. Backhouse, G. Quintero, A.J. Nunez-Selles,
M.A. Morales, Phytother. Res. 15 (2001) 18-21.
[4]
Z.-R. Lai, Y.-L. Ho, S.-C. Lai, S.-C. Huang, J.-C. Tsai, G.-J. Huang, T.-H. Huang, C.-Y.
Wang, Y.-S. Chang, Am. J. Chinese Med. 39 (2001) 1275–1290.
[5]
[6]
G. Waris, H. Ahsan, J Carcinogenesis 5 (2006) 14.
F. Conforti, S. Sosa, M. Marrelli, F. Menichini, G. A. Statti, D. Uzunov, A. Tubaro, F.
Menichini, R. Della Loggia, J. Ethnopharmacol. 116 (2008) 144–151.
G. Manda, M.T. Nechifor, T.-M. Neagu, Curr. Chem. Biol. 3 (2009) 342-366.
A. Abdel-Aziz, H.I. El-Subbagh, T. Kunieda, Bioorg. Med. Chem. 13 (2005) 4929-4935.
I.A. Yule, L.G. Czaplewski, S. Pommier, D.T. Davies, S.K. Narramore, C.W.G.
[7]
[8]
[9]

ACCEPTED MANUSCRIPT
22
Fishwick, Eur. J. Med. Chem. 86 (2014) 31-38.
[10] M.G. Mamolo, V. Falagiani, L. Vio, E. Banfi, Il Farmaco 54 (1999) 761-767.
[11] K. Chavva, S. Pillalamarri, V. Banda, S. Gautham, J. Gaddamedi, P. Yedla, C.G. Kumar,
N. Banda, Bioorg. Med. Chem. Lett. 23 (2013) 5893–5895.
[12] W. Xie, S. Xie, Y. Zhou, X. Tang, J. Liu, W. Yang, M. Qiu, Eur. J. Med. Chem. 81
(2014) 22-27.
[13] A. Perez-Rebolledo, J.D. Ayala, G.M. de Lima, N. Marchini, G. Bombieri, C. L. Zani, E.
M. Souza-Fagundes, H. Beraldo, Eur. J. Med. Chem. 40 (2005) 467–472.
[14] J.M. Hung, H.J. Arabshahi, E. Leung, J. Reynisson, D. Barker, Eur. J. Med. Chem. 86
(2014) 420-437.
[15] A.M. Sajith, K.K. Abdul Khader, N. Joshi, M.N. Reddy, M.S.A. Padusha, H.P.
Nagaswarupa, M.N. Joy, Y.D. Bodke, R.P. Karuvalam, R. Banerjee, A. Muralidharan, P.
Rajendra. Eur. J. Med. Chem. 89 (2015) 21-31.
[16] P.S. Dragovich, T.J. Prins, R. Zhou, T.O. Johnson, E.L. Brown, F.C. Maldonado, S.A.
Fuhrman, L.S. Zalman, A.K. Patick, D.A. Matthews, X. Hou, J.W. Meador, R.A. Ferre,
S.T. Worland, Bioorg. Med. Chem. Lett., 12 (2002) 733-738.
[17] N. Kumar, A. Chauhan, S. Drabu, Biomed. Pharmacother. 65 (2011) 375-380.
[18] M. Kidwai, S. Saxena, S. Rastogi, R. Venkataramanan, Curr. Med. Chem. Anti-infective
Agents 2 (4) (2003) 269-286.
[19] S.A. El-Hawash, A.E. Abdel Wahab, M.A. El-Demellawy, Arch. Pharm. (Weinheim)
339 (2006) 14-23.
[20] A.H. Abadi, T.M. Ibrahim, K.M. Abouzid, J. Lehmann, H.N. Tinsley, B.D. Gary, G.A.
Piazza, Bioorg. Med. Chem. 17 (2009) 5974–5982.
[21] M.S. Al-Said, M.S. Bashandy, S.I. Al-qasoumi, M.M. Ghorab, Eur. J. Med. Chem. 46
(2011) 137-141.
[22] H.-L. Wu, J.-K. Yuan, X.-C. Huang, F. Kou, B. Liu, F. Jia, K.-T. Wang, Y. Bai, Inorg.
Chim. Acta 390, (2012) 12-21.
[23] F. Shi, C. Li, M. Xia, K. Miao, Y. Zhao, S. Tu, W. Zheng, G. Zhang, N. Ma, Bioorg.
Med. Chem. Lett. 19, (2009) 5565-5568.
[24] C. O. Kappe, Angew Chem Int Edn. 43 (2004) 6250-6284.
[25] H.T.Y. Fahmy, S.A.F. Rostom, A.A. Bekhit, Arch. Pharm. Pharm. Med. Chem. 335

ACCEPTED MANUSCRIPT
23
(2002) 213–222.
[26] H.T.Y. Fahmy, S.A.F. Rostom, M.N.S. Saudi, J.K. Zjawiony, D. J. Robins, Arch. Pharm.
Pharm. Med. Chem. 336 (2003) 216–225.
[27] A.A. Bekhit, H.T.Y. Fahmy, S.A.F. Rostom, A.M. Baraka, Eur. J. Med. Chem. 38 (2003)
27-36.
[28] S.A.F. Rostom, M. Shalaby, M. El-Demellawy, Eur. J. Med. Chem. 38 (2003) 959–974.
[29] S.A.F. Rostom, Bioorg. Med. Chem. 14 (2006) 6475–6485.
[30] H.M. Faid-Allah, M.S. Al-Saadi, S.A.F. Rostom, H.T. Fahmy, Med. Chem. Res. 16
(2007) 300–318.
[31] M.S.M. Al-Saadi, S.A.F. Rostom, H.M. Faid Allah, Arch. Pharm. Chem. Life Sci. 341
(2008) 181–190.
[32] H.M. Faid-Allah, M.S. Al-Saadi, S.A.F. Rostom, Saudi Pharm. J. 16 (2008) 33–42.
[33] M.S.M. Al-Saadi, H.M. Faid Allah, S.A.F. Rostom, Arch. Pharm. Chem. Life Sci. 341
(2008) 424–434.
[34] S.A.F. Rostom, H.M.A. Ashour, H.A. Abd El Razik, Arch. Pharm. Chem. Life Sci. 342
(2009) 299–310.
[35] S.A.F. Rostom, G.S. Hassan, H.I. El-Subbagh, Arch. Pharm. Chem. Life Sci. 342 (2009)
584–590.
[36] S.A.F. Rostom, Bioorg. Med. Chem. 18 (2010) 2767–2776.
[37] A.A. Bekhit, H.T.Y. Fahmy, S.A.F. Rostom, A.E.A. Bekhit, Eur. J. Med. Chem. 45
(2010) 6027-6038.
[38] S.A.F. Rostom, M.H. Badr, H.A. Abd El Razik, H.M.A. Ashour, A. E. Abdel Wahab,
Arch. Pharm. Chem. Life Sci. 344 (2011) 572–587.
[39] H.M. Faidallah, S.A.F. Rostom, A.M. Asiri, K.A. Khan, M.F. Radwan, H.Z. Asfour; J.
Enz. Inhib. Med. Chem. 28 (2013) 123-130.
[40] H.M. Faidallah, K.A. Khan, S.A.F. Rostom, A.M. Asiri, J. Enz. Inhib. Med. Chem. 28
(2013) 495-508.
[41] H.M. Faidallah, S.A.F. Rostom, K.A. Khan, S.A. Basaif, J. Enz. Inhib. Med. Chem. 28
(2013) 926-935.
[42] H.M. Faidallah, S.A.F. Rostom, S.A. Basaif, M.S.T. Makki, K.A. Khan, Arch. Pharm.
Res. 36 (2013) 1354-1368.

ACCEPTED MANUSCRIPT
24
[43] S.A.F. Rostom, H.M. Faidallah, M.F. Radwan, M.H. Badr, Eur. J. Med. Chem. 76 (2014)
171-181.
[44] M.M. Ghorab, F.A. Ragab, H.I. Heiba, H.A. Youssef, M.G. El-Gazzar, Bioorg. Med.
Chem. Lett. 20 (2010) 6316-6320.
[45] D. Bandyopadhyay, S. Mukherjee, J. C. Granados, J.D. Short, B.K. Banik, Eur. J. Med.
Chem. 50 (2012) 209-215.
[46] V. Padmavathi, K. Mahesh, G. Dinneswara Reddy, A. Padmaja, Eur. J. Med. Chem. 45
(2010) 3178-3183.
[47] J.D. Bhosale, A.R. Shirolkar, U.D. Pete, C.M. Zade, D.P. Mahajan, C.D. Hadole, S.D.
Pawar, U.D. Patil, R. Dabur, R.S. Bendre, J. Pharm. Res. 7 (2013) 582-587.
[48] A. Monks, D. Scudiero, P. Skehan, R. Shoemaker, K. Paull, D. Vistica, C. Hose, J.
Jangley, P. Cronisie, A. Viagro-Wolff, M. Gray-Goodrich, H. Campell, M. Boyd, J. Natl.
Cancer Inst. 83 (1991) 757-766.
[49] M.R. Grever, S.A. Schepartz, B.A. Chabner, Seminars Oncol. 19 (1992) 622-638.
[50] M.R. Boyd, K.D. Paull, Drug Rev. Res. 34 (1995) 91-109.
[51] M.S. Blois, Nature 181 (1958) 1199–1200.

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Table and Figure Captions
Scheme 1. Synthesis of the target compounds 2-13.
Scheme 2. Synthesis of the target compounds 4-19.
Table 1. Reaction times, yields and melting points of compounds 2-19 using conventional
heating and/or microwave irradiation modes.
Table 2. In vitro growth inhibition percentage (GI%) of compounds 10, 11, 12, 14, 17 and 18
against some selected tumor cell lines at the single-dose assay.a
Table 3. In vitro growth inhibitory concentrations (GI50, ꢀM) for compounds 2-9 against some
selected tumor cell lines.a
Table 4. DPPH radical-scavenging activity of compounds 3, 6, 7, 9, 10, 11 and 12
Figure 1. General structures (A-D) for the newly synthesized compounds.
Figure 2. A schematic presentation of the reaction mechanism involved in the synthesis of
compound 11.

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Table 1. Reaction times, yields and melting points of compounds 2-19 using conventional
heating and/or microwave irradiation modes.
Cpd.
No.
R
Reaction Time
Yield (%)
M.P.
(oC)
C.H.a (h)
MWb (min.)
C.H.a (h)
MWb (min.)
C6H5
5
5
10
71
47
55
66
59
38
32
48
52
49
43
48
59
36
45
53
57
50
88
127-9
2
3
3,4-di OCH3-C6H3
10
10
10
15
15
15
15
10
10
15
15
15
NAc
20
10
20
20
76
79
81
78
65
62
71
83
74
59
66
83
NAc
73
84
76
79
117-9
122-4
126-8
132-4
146-8
156-8
150-2
152-4
207-9
201-3
192-4
136-8
>300
2-Thienyl
5
4
-
6
5
C6H5
4
6
3,4-di OCH3-C6H3
4
7
2-Thienyl
5
8
-
6
9
-
2
10
11
12
13
14
15
16
17
18
19
-
1
CH2CH3
6
CH2-C6H5
6
-
-
-
-
-
-
3
0.5
4
155-7
134-6
>300
4
6
8
209-11
a: C.H.: Conventional heating mode (h: hours).
b: MW: Microwave irradiation mode (min.: minutes).
c: NA: Not applied.

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Table 2. In vitro growth inhibition percentage (GI%) of compounds 10, 11, 12, 14, 17 and 18
against some selected tumor cell lines at the single-dose assay.a
Cell Lines
NSCLCb
10
11
12
14
17
18
c
EKVX
14.2
32.0
22.4
18.9
36.4
29.7
34.1
28.7
33.8
-
16.2
-
10.1
-
HOP92
23.6
21.5
11.3
10.4
NCI-H522
Colon Cancer
HCT-116
HCT-15
19.5
20.1
22.6
13.4
35.3
10.8
-
11.7
13.4
-
14.1
6.3
Breast Cancer
HS 578T
38.1
32.7
40.6
-
-
15.2
15.4
54.2
-
10.7
-
-
-
MDA-MB-468
T-47D
11.4
19.3
40.1
17.5
11.8
Ovarian Cancer
IGROV1
38.8
27.6
34.4
13.8
35.3
23.4
10.6
-
12.4
-
7.9
6.1
OVCAR-4
Leukemia
CCRF-CEM
HL-60(TB)
K-562
27.5
35.9
33.5
31.4
36.7
26.7
33.2
29.6
32.6
51.9
21.6
25.8
37.4
42.3
68.5
15.6
-
30.1
11.6
18.3
29.2
38.6
10.3
8.5
-
12.1
23.8
47.6
MOLT-4
20.4
12.2
RPMI-8226
Renal Cancer
A 498
22.6
31.4
31.9
51.4
41.3
15.2
-
26.5
11.9
21.7
45.8
8.6
12.3
13.8
-
14.6
5.7
ACHN
-
-
-
-
-
CAKI-1
UO-31
34.7
7.3
Melanoma
LOX IMVI
29.6
10.7
12.9
-
-
-

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SK-MEL-5
UACC-62
Prostate Cancer
PC-3
12.8
24.7
18.4
23.5
13.1
24.1
16.3
18.4
10.5
16.9
-
8.3
28.4
19.8
57.1
12.5
20.4
-
CNS Cancer
SNB-75
31.9
15.8
17.5
21.4
21.4
16.7
-
-
10.7
-
-
-
U251
a
Data obtained from NCI’s in vitro disease-oriented human tumor cell screen at 10 µM conc.
NSCLC: Non-Small Cell Lung Cancer.
b
c
Totally inactive.