Synthesis and anticancer potential of certain novel 2-oxo-N'-(2-oxoindolin- 3-ylidene)-2H-chromene-3-carbohydrazides

Article abstract of DOI:10.1016/j.ejmech.2013.09.060

Treatment of ethyl 3-hydrazinyl-3-oxopropanoate (6) with indoline-2,3-dione derivatives 7a-g gave ethyl 3-oxo-3-(2-(2-oxoindolin-3-ylidene)hydrazinyl) propanoates 8a-g which were allowed to react with the appropriate salicyaldehyde 9a and/or 9b to furnish the chromene-based hydrazones 10a-i. Compounds 10a-i displayed a significant activity against HT-29 colon cancer cell line and a moderate activity against leukemia K562 cell line. Compound 10f emerged as the most active congener toward HT-29 colon cancer cell line with IC₅₀ = 7.98 ± 0.05 μM whereas compound 10c exhibited the best antiproliferative activity against leukemia K562 cell line with IC₅₀ = 9.44 ± 0.02 μM. Moreover, compound 1e showed 87.81 ± 7% inhibition of side population (SP) HT-29 colon cancer stem cells.

Full text of DOI:10.1016/j.ejmech.2013.09.060

European Journal of Medicinal Chemistry 70 (2013) 358e363
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis and anticancer potential of certain novel 2-oxo-N’-(2-
oxoindolin-3-ylidene)-2H-chromene-3-carbohydrazides
,
b
,
a
c
a,d,
Hatem A. Abdel-Aziz a , Tilal Elsaman , Abdullah Al-Dhfyan , Mohamed I. Attia
*
*
,
Khalid A. Al-Rashood a, Abdul-Rahman M. Al-Obaid a
a Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia
b Department of Applied Organic Chemistry, National Research Center, Dokki, Cairo 12622, Egypt
c Stem Cell Therapy Program, King Faisal Specialized Hospital and Research Center, P.O. Box 3354, Riyadh 11211, Saudi Arabia
d Department of Medicinal and Pharmaceutical Chemistry, National Research Center, Dokki, Cairo 12622, Egypt
a r t i c l e i n f o
a b s t r a c t
Article history:
Treatment of ethyl 3-hydrazinyl-3-oxopropanoate (6) with indoline-2,3-dione derivatives 7aeg gave
ethyl 3-oxo-3-(2-(2-oxoindolin-3-ylidene)hydrazinyl)propanoates 8aeg which were allowed to react
with the appropriate salicyaldehyde 9a and/or 9b to furnish the chromene-based hydrazones 10aei.
Compounds 10aei displayed a significant activity against HT-29 colon cancer cell line and a moderate
activity against leukemia K562 cell line. Compound 10f emerged as the most active congener toward HT-
Received 26 April 2013
Received in revised form
7 September 2013
Accepted 24 September 2013
Available online 16 October 2013
29 colon cancer cell line with IC50 ¼ 7.98 ꢀ 0.05
proliferative activity against leukemia K562 cell line with IC50 ¼ 9.44 ꢀ 0.02
showed 87.81 ꢀ 7% inhibition of side population (SP) HT-29 colon cancer stem cells.
Ó 2013 Elsevier Masson SAS. All rights reserved.
m
M whereas compound 10c exhibited the best anti-
m
M. Moreover, compound 1e
Keywords:
Colorectal cancer
Side population (SP) cells
Chromene-2-ones
Indoline-2,3-dione
Hydrazides/hydrazones
Ring-opening
1. Introduction
membrane pumps encoded by multidrug-resistance gene 1 (MDR1)
and breast cancer-resistance gene 2 (ABCG2). SP cells resist cyto-
toxic agents and hence often survive longer. Additionally, SP cells
isolated from patients or from metastatic cell lines are enriched
with colorectal CSCs/progenitor cells or stem/progenitor cells [7].
The tumorigenic potential of SP cells has been detected in various
cancer models [8,9]. Accordingly, SP cells are considered a potential
target in cancer therapy because they are thought to be a source of
CSCs.
Colorectal cancer is the most common malignancy of the
gastrointestinal tract [10] and causes 655,000 deaths worldwide
every year [11]. Many treatment protocols have been applied to
mange colorectal cancer, but a complete cure has not been ach-
ieved. This failure to achieve a cure may be due to the presence of
colorectal CSCs that are resistant to chemotherapy and/or radiation
therapy. Therefore, it is important to use therapies that target stem
cells as well as proliferating cells to cure colorectal cancer [12].
Chromene-2-ones are widely distributed in natural products
and they display diverse biological activities. Chromene-containing
compounds exert their anti-cancer activities via inhibition of ste-
roid sulfatase, aromatase, and/or carbonic anhydrase enzymes [13e
15]. On the other hand, indoline-2,3-diones exhibit anticancer
Cancer is one of the most dreaded diseases of mankind and it is
the principal cause of mortality worldwide. Currently, one in 4
deaths in the United States is due to cancer and more than ten
million new cancer cases occur annually [1]. There are many rea-
sons for the difficulty in the control and treatment of cancer
including the clonal evaluation of cancer types and the genetic and
epigenetic instability of cancer. The recently proposed cancer stem
cell (CSC) hypothesis states that a small percentage of tumor cells
are responsible for tumor initiation and progression through un-
limited self-renewal. Numerous studies have identified CSCs in
leukemia [2], breast [3], brain [4], lung [5], colon [6], and other
cancers. Side population (SP) cells have been isolated from several
solid tumors in a technique based on the exclusion of Hoechst dye
33342 or other dyes, which occurs primarily through the activity of
* Corresponding authors. Department of Pharmaceutical Chemistry, College of
Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia.
E-mail addresses: hatem_741@yahoo.com (H.A. Abdel-Aziz), msamabd@
yahoo.com (M.I. Attia).
0223-5234/$ e see front matter Ó 2013 Elsevier Masson SAS. All rights reserved.
http://dx.doi.org/10.1016/j.ejmech.2013.09.060

H.A. Abdel-Aziz et al. / European Journal of Medicinal Chemistry 70 (2013) 358e363
359
activities by inhibiting tyrosine kinase or caspase-3/7. In 2006, the
FDA approved an indoline-2,3-dione derivative, Sunitinib
(Sutent)Ò, for the oral management of renal cell carcinoma (RCC)
and imatinib-resistant gastrointestinal stromal tumor (GIST) [16].
Additionally, we have documented the synthesis of certain novel
indoline-2,3-diones that display potent and selective anti-
proliferative activity against multidrug-resistant cancer cells
[17,18].
An examination of the literature revealed that Soliman et al.
obtained salicyaldehyde azine (3a) instead of the expected hydra-
zide 2a for the reaction of ester 1a with hydrazine hydrate (Fig. 1)
[19]. Recently, malonohydrazide (4) along with salicyaldehyde
azine (3a) were isolated from the reaction of 1a with hydrazine
hydrate and a mechanism for their formation was proposed [20]. In
the same vein, Sammour et al. reported that the reaction of ethyl 6-
bromo-2-oxo-2H-chromene-3-carboxylate (1b) with hydrazine
hydrate gave compounds 3b and 4 instead of hydrazide 2b (Fig. 1)
[21]. Additionally, Mohamed et al. [22] published the reaction of
ethyl 7-hydroxy-2-oxo-2H-chromene-3-carboxylate (1c) with hy-
drazine hydrate to give compounds 3c and 4 instead of hydrazide
2c (Fig. 1).
Interestingly, Ma et al. documented that the reaction of ethyl 7-
(diethylamino)-2-oxo-2H-chromene-3-carboxylate (1d) with hy-
drazine hydrate gave hydrazide 2d (Fig. 1) [23], which was rigor-
ously confirmed [24]. These variable findings in the reaction of
ethyl chromene-3-carboxylates 1aed with hydrazine hydrate
demonstrate that the type and position of the substituent affect
significantly on the pyran-2-one ring stability.
In view of the aforementioned premises, and in a continuation
of our interest in the synthesis of hydrazone-based compounds
with anticancer activity [17,18,25e29], we aimed to report herein
the synthesis and in vitro antiproliferative activity of the title
compounds 10aei toward a panel of cancer cell lines as well as their
inhibitory activity on side population cancer stem cells.
These findings encouraged us to establish an alternative route
for synthesis of the targeted chromene-based hydrazones 10aei
starting from ethyl 3-hydrazinyl-3-oxopropanoate (6) (Scheme 1)
instead of esters 1a,b. Consequently, treatment of 6 with the
appropriate indoline-2,3-dione derivative 7aeg in refluxing
ethanol resulted in the formation of ethyl 3-oxo-3-(2-(2-
oxoindolin-3-ylidene)hydrazinyl) propanoates 8aeg (Scheme 1).
Condensation of hydrazones 8aeg with the appropriate sali-
cyaldehyde 9a or 9b in the presence of piperidine yielded 2-oxo-N’-
(2-oxoindolin-3-ylidene)-2H-chromene-3-carbohydrazides 10aei.
IR spectra of compounds 10aei revealed three carbonyl absorption
bands around 1700 cmꢁ1 as well as two NH absorption bands in the
3450e3100 cmꢁ1 region. Additionally, their 1H NMR spectra
showed two D2O-exchangeable signals in the region
d 9.07e11.3
and 11.0e11.95 due to NH of isatin and hydrazone functionalities,
d
respectively.
2.2. In vitro antiproliferative activity
In vitro antiproliferative activity of compounds 10aei against
leukemia K562, breast MDA-MB-468 and colon HT-29 cell lines is
displayed in Table 1. Compound 10c (R/R1 ¼ H/Cl) exhibited the
highest antiproliferative activity toward leukemia K562 cell line with
IC50
¼
9.44
ꢀ
0.02
m
M
followed by compounds 10d
M) which
(IC50 ¼ 12.08 ꢀ 0.01
mM) and 10f (IC50 ¼ 12.99 ꢀ 0.1
m
contain bromine and methoxy substituents at five position of indo-
line-2,3-dione moiety, respectively. The antiproliferative activity of
the other synthesized congeners is in the following decreasing order:
10a (IC50 ¼ 25.5 ꢀ 08
mM) > 10b > 10i > 10g > 10e > 10h
M). On the other hand, compound 10h
(IC50 ¼ 36.99 ꢀ 0.09
m
(IC50 ¼ 15.48 ꢀ 0.02
mM) is the most active candidate among all the
synthesized compounds 10aei against breast MDA-MB-468 cell line
while compound 10f exhibited the weakest antiproliferative activity
with IC50 ¼ 43.97 ꢀ 0.17
mM.
The best in vitro antiproliferative profile of compounds 10aei is
showed against colon HT-29 cancer cell line where compound 10f is
2. Results and discussion
the most active congener with IC50 ¼ 7.98 ꢀ 0.05
mM. Compound 10f
has un-substituted chromene moiety and a substituent, methoxy
group, endowed with negative inductive and positive mesomeric
effects at the five position of indoline-2,3-dione nucleus. Com-
pounds 1e (R/R1 ¼ H/Me), 1g (R/R1 ¼ H/NO2) and 1d (R/R1 ¼ H/Br)
exhibited a comparable biological activity to that of compound 10f
2.1. Chemistry
Hydrazides are versatile tools for the construction of several
functionalized heterocycles with interesting biological activities
[30e36]. Therefore, the synthesis of hydrazide-containing com-
pounds has drawn considerable interest [37,38]. The common route
for the synthesis of chromene-based hydrazones is the reaction of
the appropriate hydrazide 2 with indoline-2,3-diones. Many re-
ports indicate that hydrazide 2a is produced from the reaction of
ester 1a with hydrazine hydrate (Fig. 1) and that 2a can be used as a
key starting material in the synthesis of several heterocyclic com-
pounds and hydrazones [39e48]. However, the ring opening of
chromene-2-ones under moderate conditions has been reported
where the aliphatic pyran-2-one ring in the chromene system is
highly reactive. It undergoes ring opening at the lactone acyl center
under nucleophilic attack as well as under nucleophilic conjugate
addition at the carbonecarbon double bond [49].
with IC50 ¼ 8.42 ꢀ 0.01, 9 ꢀ 0.01 and 10.05 ꢀ 0.03
mM, respectively.
The decreasing order of antiproliferative activity of the rest com-
pounds toward colon HT-29 cancer cell line is as follows: 1h
(IC50 ¼ 12.25 ꢀ 0.04
mM) > 1b > 1i > 1a > 1c (IC50 ¼ 27.1 ꢀ0.46
mM).
2.3. Side population cancer stem cells inhibitor activity
The inhibitory activity of the target compounds 10aei against
side population colon HT-29 cancer stem cells was performed at
10
m
M (Table 2 and Fig. 2). Compound 1e (R/R1 ¼ H/Me) is the most
active inhibitor displaying 87.8 ꢀ 7% inhibition as compared with
the reference compound, Verapamil, which showed 95 ꢀ 8%
R1
O
O
O
O
R2
H2NHN
R1
R2
O
O
R1
R2
R1
R2
N
OEt
NHNH2
N
NHNH2
O
1a-d
O
2a-d
OH
3a-d
HO
4
1-3: a, R1 = R2 = H; b, R1 = Br, R2 = H; c, R1 = H, R2 = OH; d, R1 = H, R2 = N(Et)2
Fig. 1. The structure of compounds 1e3 (aed) and 4.

360
H.A. Abdel-Aziz et al. / European Journal of Medicinal Chemistry 70 (2013) 358e363
O
R1
R1
O
O
O
N
H
O
O
O
O
N
7a-g
EtO
N
H
NH2NH2.H2O
NH2
EtO
N
H
EtO
OEt
NH
5
O
6
8a-g
R
CHO
OH
R1
H
10
a
b
c
d
e
f
R
Pip.
H
H
H
H
H
H
H
Br
Br
F
9a, b
Cl
Br
Me
R1
O
O
MeO
NO2
H
R
N
g
h
i
N
H
NH
F
O
O
10a-i
Scheme 1. Synthesis of hydrazones 10aei.
inhibition. On the other hand, compounds 10f (R/R1 ¼ H/MeO) and
10i (R/R1 ¼ Br/F) increased the side population, while the rest
compounds showed variable inhibitions varied from 78.05 ꢀ 7% for
10h (Br/R1 ¼ Br/F) to 46.36 ꢀ 14% for 10a (R/R1 ¼ H/H).
determined on a Gallenkamp melting point apparatus and are
uncorrected. NMR Spectra were scanned in DMSO-d6 on a Jeol NMR
spectrometer operating at 400 MHz for 1H and 100 MHz for 13C.
Chemical shifts are expressed in d-values (ppm) relative to TMS as
an internal standard. Coupling constants (J) are expressed in Hz.
D2O was added to confirm the exchangeable protons. Mass spectra
were measured on an Agilent Triple Quadrupole 6410 QQQ LC/MS
equipped with an ESI (electrospray ionization) source.
3. Conclusion
Synthesis and in vitro antiproliferative activity of certain new 2-
oxo-N0-(2-oxoindolin-5-substituted-3-ylidene)-2H-chromene-3-
carbohydrazides 10aei toward leukemia K562, breast MDA-MB-
468 and colon HT-29 cell lines are reported. The title compounds
10aei exhibited good antiproliferative profile against colon HT-29
cell line where compounds 10f and 10e emerged as the most
4.1.2. General procedure for the synthesis of ethyl 3-oxo-3-(2-(2-
oxoindolin-3-ylidene)hydrazinyl)propanoates 8aeg
Ethyl 3-hydrazinyl-3-oxopropanoate (6) was prepared from
diethyl malonate (5) and hydrazine hydrate according to the re-
ported method (m.p. 68e69 ꢂC) [50]. A solution of compound 6
(1.46 g, 10 mmol) and the appropriate indoline-2,3-dione 7aeg
(10 mmol) in absolute ethanol (30 mL) was refluxed for 1 h and
then left to cool. The solid formed was collected by filtration,
washed with ethanol to give hydrazones 8aeg which were used for
the next step without any further purification.
active candidates with IC50 ¼ 7.98 ꢀ 0.05 and 8.42 ꢀ 0.01
mM,
respectively. Additionally, the target compounds 10aei displayed a
moderate inhibitory activity for side population (SP) colon HT-29
cancer stem cells. Compound 10e showed 87.81 ꢀ 7% inhibition of
SP colon HT-29 cancer stem cells where the reference compound,
verapamil, exhibited 95 ꢀ 8% inhibition.
4.1.3. General procedure for the synthesis of 2-oxo-N0-(2-
4. Experimental
oxoindolin-3-ylidene)-2H-chromene-3-carbohydrazides 10aei
To a solution of the appropriate hydrazone 8aeg (1 mmol) and
salicyaldehyde (9a) or 5-bromosalicyaldehyde (9b) (1 mmol) in
absolute ethanol (25 mL), a catalytic amount of piperidine (0.3 mL)
was added. The reaction mixture was refluxed for 1 h. The formed
precipitate was filtered off, washed with ethanol, dried and
4.1. Chemistry
4.1.1. General
Infrared (IR) Spectra were recorded as KBr disks using the Perkin
Elmer FT-IR Spectrum BX apparatus. Melting points were
Table 1
In vitro antiproliferative activity for compounds 10aei on K562 chronic myeloge-
nous leukemia, MDA-MB-468 breast cancer and HT-29 colon cancer cells.
Table 2
(m
M)a
Side population inhibition activity on HT-29 colon cancer stem cells for compounds
10aei.
Compound 10
R
R1
IC50
Leukemia K562 Breast MDA-MB-468 Colon HT-29
Compound 10
R
R1
Side population
Inhibition (%) ꢀ SD at 10
a
b
c
d
e
f
g
h
i
H
H
H
H
H
H
H
Br
Br
H
F
Cl
Br
Me
25.5 ꢀ 08
17.2 ꢀ 0.01
17.52 ꢀ 0.01
16.45 ꢀ 0.11
27.97 ꢀ 0.02
18.21 ꢀ 0.04
43.97 ꢀ 0.17
31.01 ꢀ 0.15
15.48 ꢀ 0.02
40.25 ꢀ 0.24
16.7 ꢀ 0.3
16.25 ꢀ 0.5
27.1 ꢀ 0.46
10.05 ꢀ 0.3
8.42 ꢀ 0.01
7.98 ꢀ 0.05
9 ꢀ 0.01
mM
25.77 ꢀ 0.07
9.44 ꢀ 0.02
12.99 ꢀ 0.13
35.01 ꢀ 0.43
a
b
c
d
e
f
g
h
H
H
H
H
H
H
H
Br
Br
H
F
Cl
Br
Me
MeO
NO2
H
46.36 ꢀ 14
68.36 ꢀ 8
73.81 ꢀ 7
73.81 ꢀ 7
87.81 ꢀ 7
Ina
58.54 ꢀ 7
78.05 ꢀ 7
Ina
MeO 12.08 ꢀ 0.01
NO2 27.64 ꢀ 0.48
H
F
36.99 ꢀ 0.09
27.4 ꢀ 0.22
12.25 ꢀ 0.4
16.58 ꢀ 0.7
a
IC50: concentration of the compound (mM) producing 50% cell growth inhibition
i
F
after 48 h of compound exposure, as determined by the WST-1 assay. Each exper-
iment was run at least two times, and the results are presented as average values
ꢀS.D.
Verapamil
95 ꢀ 8
a
Increase the side population cancer stem cells.

H.A. Abdel-Aziz et al. / European Journal of Medicinal Chemistry 70 (2013) 358e363
361
Fig. 2. The histograms of flow cytometric identification of side population cells in HT-29 colon cancer cells for compounds 10aei and verapamil.
recrystallized from EtOH/DMF to give the title carbohydrazides
10aei.
4.1.3.2. N0-(5-Fluoro-2-oxoindolin-3-ylidene)-2-oxo-2H-chromene-
3-carbohydrazide (10b). Yield (62%); mp 310e312 ꢂC; IR (KBr):
3450e3197 (2NH), 1700 (3C]O) cmꢁ1 1H NMR (DMSO-d6):
6.93e9.14 (m, 8H, 7ArHs and 1H of pyran), 11.19 (s, D2O exch., 1H,
NH of isatin), 11.89 (s, D2O exch., 1H, NH of hydrazide); ESI MS m/z:
352.1 [M þ 1]þ.
n
;
4.1.3.1. 2-Oxo-N0-(2-oxoindolin-3-ylidene)-2H-Chromene-3-
d
carbohydrazide (10a). Yield (69%); mp 318e320 ꢂC; IR (KBr):
n
3450e3164 (2NH), 1705 (3C]O) cmꢁ1
6.93e9.13 (m, 9H, 8ArHs and 1H of pyran), 11.13 (s, D2O exch., 1H,
NH of isatin), 11.89 (s, D2O exch., 1H, NH of hydrazide); 13C NMR
(DMSO-d6): 116.8, 116.9, 118.9, 119.7, 120.2, 125.6, 127.8, 128.1,
;
1H NMR (DMSO-d6):
d
4.1.3.3. N0-(5-Chloro-2-oxoindolin-3-ylidene)-2-oxo-2H-chromene-
d
3-carbohydrazide (10c). Yield (70%); mp > 360 ꢂC; IR (KBr):
n
129.6, 130.9, 131.0, 134.5, 136.8, 141.4, 150.1, 154.5, 161.6, 165.8; ESI
3450e3196 (2NH), 1734e1654 (3C]O) cmꢁ1; 1H NMR (DMSO-d6):
d 6.93e9.12 (m, 8H, 7ArHs and 1H of pyran), 11.13 (s, D2O exch., 1H,
MS m/z: 334.1 [M þ 1]þ, 356.1 [M þ 23]þ.

362
H.A. Abdel-Aziz et al. / European Journal of Medicinal Chemistry 70 (2013) 358e363
NH of isatin), 11.89 (s, D2O exch., 1H, NH of hydrazide); 13C NMR
(DMSO-d6): 116.8, 117.0, 118.9, 119.9, 125.8, 127.2, 127.9, 129.4,
containing the desired concentrations of the test compounds. After
d
48 h, 10 ml of the WST-1 reagent were added to each well and the
130.8, 133.5, 134.5, 134.9, 141.4, 145.2, 149.9, 154.5, 161.6, 165.3; ESI
plates were re-incubated for 4 h at 37 ꢂC. The amount of formazan
MS m/z: 368.1 [M þ 1]þ, 390.1 [M þ 23]þ.
was quantified using ELISA reader at 450 nm.
4.1.3.4. N0-(5-Bromo-2-oxoindolin-3-ylidene)-2-oxo-2H-chromene-
4.3. Side population cancer stem cells inhibitor activity
3-carbohydrazide (10d). Yield (70%); mp 302e304 ꢂC; IR (KBr):
n
3442e3100 (2NH), 1701 (3C]O) cmꢁ1
;
1H NMR (DMSO-d6):
The side population cancer stem cells inhibitor activity of the
test compounds 10aei was evaluated at Stem Cell Therapy Pro-
gram, King Faisal Specialized Hospital and Research Center, P.O.
3354, Riyadh 11211, Saudi Arabia. Side population staining was
performed by using VybrantÒ DyeCycle Violet Assay Kit (Molecular
Probe) following the manufacturer’s recommendations. Briefly,
1*106 of HT-29 colon cancer cells suspended in 1 mL of DMEM
d
6.97e8.93 (m, 8H, 7ArHs and 1H of pyran), 11.13 (s, D2O exch., 1H,
NH of isatin), 11.90 (s, D2O exch., 1H, NH of hydrazide); ESI MS m/z:
412.1 [M]þ.
4.1.3.5. N0-(5-Methyl-2-oxoindolin-3-ylidene)-2-oxo-2H-chromene-
3-carbohydrazide (10e). Yield (73%); mp 310e312 ꢂC; IR (KBr):
n
3400e3197 (2NH), 1700 (3C]O) cmꢁ1; 1H NMR (DMSO-d6):
(s, 3H, CH3), 6.89e8.92 (m, 8H, 7ArHs and 1H of pyran),11.13 (s, D2O
exch., 1H, NH of isatin),11.90 (s, D2O exch., 1H, NH of hydrazide); 13
NMR (DMSO-d6): 20.5, 116.8, 116.9, 118.9, 119.9, 125.8, 127.2, 127.9,
d 2.48
medium and 10
incubated for 90 min in 37 ꢂC. Cells were centrifuged and washed
by the medium. As a control, cells were treated with 100
m
M of VybrantÒ DyeCycle Violet were added and
C
m
M
d
Verapamil (sigma), which blocks the action of the transporter
responsible for dye exclusion. The analyses were performed on a
FACS LSRII (BD Biosciences). Debris and cell clusters were excluded
during side-scatter and forward-scatter analyses. Results are given
in Table 2 and Fig. 2.
129.4, 130.9, 134.5, 134.9, 141.4, 145.2, 148.2, 149.9, 154.5, 161.6,
164.8; ESI MS m/z: 348.1 [M þ 1]þ, 370.1 [M þ 23]þ.
4.1.3.6. N0-(5-Methoxy-2-oxoindolin-3-ylidene)-2-oxo-2H-chro-
mene-3-carbohydrazide (10f). Yield (60%); mp > 360 ꢂC; IR (KBr):
n
3380e3197 (2NH), 1701 (3C]O) cmꢁ1; 1H NMR (DMSO-d6):
d 3.34
Acknowledgments
(s, 3H, OCH3), 6.95e8.93 (m, 8H, 7ArHs and 1H of pyran), 11.13 (s,
D2O exch., 1H, NH of isatin), 11.89 (s, D2O exch., 1H, NH of hydra-
zide); ESI MS m/z: 364.1 [M þ 1]þ, 386.2 [M þ 23]þ.
The authors would like to extend their sincere appreciation to
the Deanship of Scientific Research at King Saud University for its
funding of this research through the Research Group Project no.
RGP-VPP-321.
4.1.3.7. N0-(5-Nitro-2-oxoindolin-3-ylidene)-2-oxo-2H-chromene-3-
carbohydrazide (10g). Yield (62%); mp 320e322 ꢂC; IR (KBr):
n
3300e3197 (2NH), 1700 (3C]O) cmꢁ1
;
1H NMR (DMSO-d6):
Appendix A. Supplementary data
d
6.95e8.93 (m, 8H, 7ArHs and 1H of pyran), 11.13 (s, D2O exch., 1H,
NH of isatin), 11.95 (s, D2O exch., 1H, NH of hydrazide); ESI MS m/z:
379.2 [M þ 1]þ, 401.1 [M þ 23]þ.
Supplementary data related to this article can be found at http://
dx.doi.org/10.1016/j.ejmech.2013.09.060.
4.1.3.8. 6-Bromo-2-oxo-N0-(2-oxoindolin-3-ylidene)-2H-chromene-
References
3-carbohydrazide (10h). Yield (74%); mp 308e310 ꢂC; IR (KBr):
n
3446e3229 (2NH), 1707 (3C]O) cmꢁ1
;
1H NMR (DMSO-d6):
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