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Synthesis and Pharmacological Evaluation of Novel Coumarin Derivatives

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DOI: 10.26452/ijrps.v11i1.1908

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 Sanaryh Mohammed Al-Awad et al., Int. J. Res. Pharm. Sci., 2020, 11(1), 865-874

ORIGINAL ARTICLE

INTERNATIONAL JOURNAL OF RESEARCH IN

PHARMACEUTICAL SCIENCES
Published by JK Welfare & Pharmascope Foundation Journal Home Page: www.pharmascope.org/ijrps

Synthesis and Pharmacological Evaluation of Novel Coumarin

Derivatives
Sanaryh Mohammed Al-awad*1 , Leaqaa Abdalredha raheem1 , Ausama Ayob Jaccob2
1
Department of pharmaceutical chemistry, College of Pharmacy, University of Basrah, Iraq
2
Department of pharmacology and toxicology, College of Pharmacy, University of Basrah, Iraq

Article History: ABSTRACT

Received on: 07.09.2019 The current work focuses on new architecture, synthesis of coumarin-
Revised on: 10.12.2019 oxadiazole hybrid derivative products as both these (coumarin ring and oxa-
Accepted on: 24.12.2019 diazole) have a wide variety of biological behavior, Compounds containing
the nucleus of coumarin (2H-1-benzopyran-2-one) are an interesting class of
Keywords:
hetero cycles which hold an important role in the ield of natural ingredients
and synthetic organic chemistry. It has been exciting medicinal chemists to
Coumarin derivatives, study native coumarins or synthetic analogs for their application for decades.
1 3 4- Derivatives of And they can be further modi ied to synthesize more effective and potent
Oxadiazole, drugs. Compounds have been characterized by spectrophotometry of physic-
Biological activities, ochemical properties and their structures veri ied by infrared spectroscopy
Coumarin- Oxadiazole (FTIR) and nuclear magnetic resonance (1H-NMR) Such new derivatives of
derivatives coumarinyl-oxadiazole was quali ied to estimate the lethal dose, anticancer,
anticoagulant and antioxidant activity. Their pharmacological properties
depend on their pattern of substitution, compound S4F proved signi icant
anticoagulant activity in concentration (50, 100, 200 mg/ml) similar for hep-
arin, and monitor the coagulation effect on plasma, while compound S4 CO
give signi icant anticancer activity against MCF-7 a breast cancer cell. Spe-
ci ic compounds have strong antioxidants with the effective action of radical
scavengers; the S4Cl compound with IC50 1.49 is the most potent antiox-
idant activity note. Basically, all the formulations tested reported satisfac-
tory behavior. The review shows that varieties of coumarin derivatives have
synthesized and shown anti-cancer, antioxidant and anti-coagulant potentials.
These derivatives synthesis and its biological assay can be further modi ied in
the future to improve the anti-cancer, anti-oxidant and anticoagulant poten-
tials of the versatile coumarin nucleus.

*
Corresponding Author INTRODUCTION
Name: Sanaryh Mohammed Al-awad Cumarin derivatives have a wide range of biologi-
Phone: 07801022218 cal functions. Diverse heterocyclic compounds con-
Email: [email protected] sisting of coumarins are among the most impor-
tant groups of heterocycles holding a leading posi-
ISSN: 0975-7538 tion in synthetic and therapeutic chemistry, lead-
ing to their different applications as anti-oxidant,
DOI: https://doi.org/10.26452/ijrps.v11i1.1908 reduce in lammation, bactericidal, antitumor and
blood thinner compound (Manojkumar et al., 2009).
Production and Hosted by
In addition to 1,3,4-oxadiazoles display evident anti-
Pharmascope.org coagulant (Jeong et al., 2004; Saibara et al., 2003)
© 2020 | All rights reserved. and anti-cancer properties (Lin et al., 2007; Oza
et al., 2012).

© International Journal of Research in Pharmaceutical Sciences 865

 Sanaryh Mohammed Al-Awad et al., Int. J. Res. Pharm. Sci., 2020, 11(1), 865-874

Coumarins have attracted researchers to work on Glacial acetic acid, Thomas baker, India/ Heparin,
this moiety, which is instrumental in the creation of HAVER (25,000 IU/5ml), Canada/ Blood plasma,
new coumarin compounds, a large spectrum of bio- From the researcher (Sanaryh.M)/ DMSO Merck,
logical activity and its effectiveness as valuable syn- Germany, Ascorbic acid/ Sigma, Aldrich and 1,1-
thons, while oxadiazoles have a great contribution to diphenyl-2-picrylhydrazyl (DPPH)/Sigma, Aldrich.
the development of heterocyclic chemistry. Numer- Instrumentation Condition
ous oxadiazoles have synthesized and subjected to
biological screening; the results have increased their The FT-IR8400S spectrophotometer(SHIMADZU /
importance because of potential activities and use Japan) was reported the infrared spectrum as KBr
in different ields of daily life. To date, our researchwavelengths 1 HNMR (Proton nuclear magnetic res-
is based on the new synthesis of coumarinyl -1, 3, onance) spectra was calculated by a College of Sci-
4-oxadiazole derivatives, considering it has a large ence and Technology – Iran on a Bruker Ultra shield
variety of medical and industrial uses. 499 MHz spectrometer (Switzerland) system on
Dimethyl - sulphoxide (DMSO-d6). Chemical shift-
The novel products prepared from coumarin - ing of hydrogen atoms are measured in proportional
3-carboxylic acid through synthesis hydrazide– parts per million (ppm) relative to the internal norm
hydrazine compound (CO-NHN=CH) as an interme- of tetramethylsilane.
diate product. By cyclization of that intermediate
to give 1,3,4-oxadiazole ring moiety at 3 position TLC (Analytical thin-layer chromatography) was
in coumarin ring, necessary to get effective bind- conducted on silica gel coated plates (Merck 60
ing with different enzymes and receptors in biolog- F254, 0.25 mm), which were visualized under 254
ical systems eliciting an array of bioactivities spec- nm of ultraviolet or iodine mist.
trum including anticancer (Nasr et al., 2018), anti- Compounds Synthesis
coagulant (Rishavy et al., 2018), and antioxidant (Al- Methyl 2-oxo-2H-chromene-3-carboxylate (S )
1
Majedy et al., 2016).
In100 ml broad bottom lask linked to a con-
We prepared this work to synthesis and design four denser for re lux, coumarin -3-carboxylic acid (2g,
novel coumarin derivatives substituted at site 3 by 1mol)was solubilized in20ml absolute methanol,
oxadiazole ring and rationalize the pharmacological and then 3 drops of sulphuric acid were applied
activity such as antioxidant, anticoagulant and anti- and the re lux system heated for 7 hours., cooling
cancer to observe clear analysis into the relation- the reaction and evaporated the mixture to dryness
ship between structure and behavior of these com- and the subsequent mixture of reaction extracted
pounds. with the ethyl acetate, then added 5% bicarbonate
Aim of the study of sodium until the solution becomes basic.
We based the present study on a synthesis of The inal product, as shown in (Scheme 1), was sep-
coumarin heterocyclic compounds and then phar- arated by 25 ml dichloromethane using separating
macological study for this derivative such as anti- funnel (Manvar et al., 2008).
cancer, antioxidant and anticoagulant activity, and Off-white needle-like crystals.Production of 75%.
to provide the development of substituted coumarin M.P = 113◦ C, RF=0.75 (Ethyl acetate n-Hexane: 3:7);
nucleus to give a potent, bene icial product. The IR (cm−1 ): 2933 (Aliphatic, C-H.), 3055 (Aro-
matic C-H,), 1745.5 (Ester C=O,), 1683.8 (Lactone,
MATERIALS AND METHODS C = O.), 1610.53 (Alkene C=C,), 1567.2 (Aromatic
C=C,).
Reagents and chemicals
2-oxo-2H-chromene-3-carbohydrazide (S2 )
Coumarone -3-carboxylic acid, Sigma–Aldrich Ger-
man. /. Methanol, sigma–Aldrich, German / In 100 ml lat bottom round lask, coumarin ester
conc. Sulphuric acid 99%, Merk, German. / Ethyl S1 (0.1mol) dissolved in 10 ml ethanol, then added
acetate, Alpha Chemika, India / Hexane, sigma– (0.5 mol) hydrazine hydrate (98%), re luxed the
Aldrich German. / Hydrochloric acid, Merk Ger- mixture for 12hrs, cooled at room temperature,
man / Absolute ethanol, sigma–Aldrich, German. / the reaction combination remains precipitate to the
Hydrazine hydrate 80%, ALPHA Company, India./ next day.
Chloroform, SDFCL, India./Aromatic aldehyde (Ben- The solid product, as shown in (Scheme 2) il-
zyaldehyde), 4-chlorobenzyladehyde, 4- loro ben- tered separately, and recrystallized with ethanol
zylaldehyde, Merck, Germany /. 4- methoxy ben- to give off white glittery crystals. (Manvar et al.,
zyldehyde, BDH, England /Acetic anhydride, Merck, 2008).White shiny crystals; yield (65-70) %. M.P
Germany/ pyridine, Hayashi Pure Chemical, Japan/ = 90-93◦ C; RF=0.66 (Ethyl acetate n-Hexane: 6:4).

866 © International Journal of Research in Pharmaceutical Sciences

 Sanaryh Mohammed Al-Awad et al., Int. J. Res. Pharm. Sci., 2020, 11(1), 865-874

Figure 1: Synthesis of coumarin -1, 3, 4- Oxadiazole derivative

The IR (cm−1 ): 2933 (Aliphatic C-H,), 3043 (Aro- of the reaction to provide the yellow component
matic C-H,), 3386, 3290 (Hydrazide NH-NH2 ), 1745 recrystallized with ethanol.
(Lactone C =O), 1614 (C=O,CONH), 1608 (Alkene All derivatives structures were reported by physi-
C=C,), 1573 (Aromatic C=C,). cal and chemical properties (Color, M.P, as well as
N’-benzylidene-2-oxo-2H-chromene-3- TLC system ( Ethyl acetate: n-hexane::7:3) and also
carbohydrazide synthesis (S3 ) con irmed by FT-IR spectrometry (Berthomieu and
Hienerwadel, 2009; Manvar et al., 2008).
In 100 ml lat bottom round lask, A
mixture of S2 (0.01 mol) and aromatic N’-benzylidene-2-oxo-2H-chromene-3-
aldehydes (0.01mol, benzaldehyde, 4- carbohydrazide (S3 H)
loroubenzaldehyde, 4-chlorobenzenedehyde, Yellowish powder as shown in (Scheme 3) ;
and 4-methoxybenzaldehyde was re luxed in abso- M.P.=85◦ C ; R.F=0.55 ; The IR ( cm−1 ): 3079 (C-H),
lute ethanol and 2-3 drops of glacial acetic acid, for 2970 (As C-H ), 2865 (Sy C-H ), 3437 (Secondary
4 hours ethanol was evaporated after inalization amine N-H), 1687 (C=N), 1620 (Amide C=O), 1766

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(Lactone C=O), 1483 (C=C), 1269 (C-O). , 1600(C=C),1199(C-N), 1253 (C-O). 1 H NMR (499
N’-(4- luorobenzylidene)-2-oxo-2H-chromene- MHz, ppm, DMSO-d6 ): 8.66 ppm (s,1H.Oxa.ring),
3-carbohydrazide (S3 F) 2.36 ppm (s, H, CH3) group and rang 7.26-8.3 ppm
(d., t. Ar-H system).
Pale Yellow powder as shown in (Scheme 4); M.P
=137◦ C; RF=0.5; The IR ( cm−1 ): 3079 (C-H ), 2970 3-(4-acetyl-5-(4-chlorophenyl)-4, 5-dihydro-1,
(As C -H), 2865 (Sy C-H) , 3437 (Secondary amine N- 3, 4-oxadiazol-2-yl)-2H-Chromen-2-one (S4 Cl)
H ), 1687 (C=N ), 1620 (Amide C=O,), 1766 (Lactone The light yellow powder, as shown in (Scheme 9),
C=O) , 1483 (C=C), 1269 (C-O). Yield (33%), M.P. =162◦ C, RF=0.05. The IR
−1
N’- (4-chlorobenzylidene)-2-oxo-2H-chromene- (cm ): 3113 (Aromatic C-H), 2935 (Aliphatic C-H),
3-carbohydrazide (S3 Cl) 1768 (Lacton C=O), 1676 (C=N), 1620 (C=O), 1600
(C=C),1199 (C-N), 1253 (C-O).
Light yellow powder as shown in (Scheme 5) ; M.P. = 1
158 ˚C ; RF= 0.62 ; The IR (cm−1 ): 3079 (C-H), 2970 H NMR (499 MHz, ppm, DMSO-d6 ): 8.66 ppm (s,1H.
(As C-H) , 2865 (Sy C-H), 3437 (Secondary amine N- Oxa. Ring), 2.36 ppm (s, H, CH3) group. and rang
H), 1687 (C=N), 1620 (Amide C=O), 1766 ( Lactone 7.26-8.3 ppm (d., t., Ar-H system)
C=O), 1483 (C=C), 1269 (C-O). 3-(4-acetyl-5-(4-methoxyphenyl)-4, 5-dihydro-
N’-(4-methoxybenzylidene)-2-oxo-2H- 1, 3, 4-oxadiazol-2-yl)-2H-chromen-2-one
chromene-3-carbohydrazide (S3 CO). (S 4 CO)
Pale yellow crystal as shown in (Scheme 6); M.P. White crystals as shown in (Scheme 10), Yield
=143◦ C ; R F = 0.6; The IR (cm−1 ): 3079 (C-H), 2970 (67%), M.P.=136 ˚C , RF=0.52; The IR ( cm–1 ):
(As C-H), 2865 (Sy C-H ), 3437 (Secondary amine N- 3113(Aromatic C-H ), 2935 (Aliphatic C-H), 1768
H), 1687 (C=N), 1620 (Amide C=O), 1766 (Lactone (Lacton C=O) , 1676 (C=N), 1620 (C=O), 1600 (C=C),
C=O), 1483 (C=C), 1269 (C-O). 1199(C-N), 1253 (C-O).
1
Coumarin1, 3, 4-Oxadiazole derivatives (S4 ) H NMR (499 MHz, ppm, DMSO-d6 ): 8.66 ppm (s, 1H.
Oxa. Ring), 2.36 ppm (s, H, CH3) group. Rang 7.26-
The reaction mixture of S3 compounds (0.5
8.3 ppm (d, t, Ar-HH system)
g,0.01mol) and excess acetic anhydride (10 ml,
0.01 mol) with 4-5 drops of pyridine added in 100
ml round lask, then re luxed for 2 hours.
The mixture of the reaction was left during the night
at room temperature when the yellowish-like solid
mass was isolated and obtained by iltration and
washed with water.
Scheme 1: S1
Ethanol recrystallized the product to get the desired
product. (Aa and Mg, 2015). FTIR spectrometry,
1HNMR (Macomber and Harbison, 1999) con irmed
structures of all oxadiazole derivatives and TLC (n-
hexane: ethyl acetate/2:8) reported structures.
3-(4-acetyl-5 phenyl-4, 5-dihydro-1, 3, 4-
oxadiazol-2-yl)-2H- Chromen-2-one (S4 H)
Light yellow powder as shown in (Scheme 7); Yield
(28 %)., M.P. =109◦ C, RF.= 0.4 ; The IR ( cm-1 ): 3113 Scheme 2: S2
(Aromatic C-H), 2935(Aliphatic C-H), 1768 ( Lacton
C= O), 1676 (C= N), 1620 (C= O), 1600 (C= C), 1199
(C-N), 1253 (C-O), 1H NMR (499 MHz, ppm, DMSO-
d6): 8.66 ppm (s,1H. Oxa.ring), 2.36 ppm (s, H, CH3)
and range 7.26-8.3 ppm (d, t, Ar-H system).
3-(4-acetyl-5-(4- luorophenyl)-4, 5-dihydro-1,
3, 4-oxadiazol-2-yl)-2H-Chromen-2-one (S4 F)
Very light yellow powder as shown in (Scheme 8),
Yield (63%), M.P. =103◦ C, RF = 0.49 ; The IR (
cm−1 ) : 3113 (Aromatic C-H), 2935 (AliphaticC- Scheme 3: S3H
H) , 1768 (Lacton C=O), 1676 (C=N), 1620 (C=O)

868 © International Journal of Research in Pharmaceutical Sciences

 Sanaryh Mohammed Al-Awad et al., Int. J. Res. Pharm. Sci., 2020, 11(1), 865-874

Scheme 4: S3F

Scheme 9: S4Cl

Scheme 5: S3Cl

Scheme 10: S4CO

Biological assay
LD50 measurement
With a slight adjustment on the general procedure,
the median lethal dose (LD50 ) ) of four novel syn-
thesis compounds was estimated in mice. Initially,
the test required four animals for each tested com-
Scheme 6: S3CO
pound (S4 H, S4 F, S4 Cl, and S 4 CO). 100, 200, 300, and
400 mg/kg, gavage for each mouse as the irst trial
for LD50 measurement. We carried out the same
concentrations for each tested drugs. We observed
no mortality at these concentrations. The second
stage then started involves administration of 500,
600, 700, and 800 mg/kg gavage for each mice as a
second trial for LD50 measurement. The LD50 deter-
mined for both S4F and Cl but no mortality was
noticed for LD50 measurements in S4 H and S4 CO
coumarin derivatives, so we started the 3rd stage,
900, 1000, 1100, and 1200 mg/kg gavage for each
Scheme 7: S4H mouse I did conformity investigations for each lethal
dose in the four tested compounds. LD50 measured
according to the following equation Chinedu et al.
(2013)
LD50 = [M0+M1] ÷ 2 (M0= Highest calculated dose
were no mortality observed)
(M1=Lowest calculated dose were mortality
observed)
Antioxidant action
Free radical compound hunting activity of coumarin
derivatives (S4 H, S4 F, S4 Cl, S4 CO) measured by DPPH
Scheme 8: S4F with few modi ication of dependable procedure,
(0.1 mmol) methanol-prepared DPPH solution, (0.1

© International Journal of Research in Pharmaceutical Sciences 869

 Sanaryh Mohammed Al-Awad et al., Int. J. Res. Pharm. Sci., 2020, 11(1), 865-874

ml) added to (0.4 ml) speci ic concentration sam- shake and incubated in 5 percent CO2 incubator for
ple solutions (50,100,150,200,250 µg/ml) and (0.5 24 hours at 37 ◦ Supporting cell association, pro-
ml) methanol solution. Then shake the combina- liferation and convergence of monolayers. After 24
tion and sit in the dark for 30 minutes at room hours, we assessed cell viability for therapy by elim-
temperature. Ascorbic acid with the same sam- inating the medium, adding MTT (dye) solution for
ple concentrations and the identical procedure used 100 µl of 2mg / ml and incubating at 37 C for 2 hours.
as a positive control. We measured the absorben- Solubilize the remaining crystals in the wells after
cies at (516 nanometers) using the spectrophotome- extracting the MTT solution by adding 90 µl of DMSO
ter (ChemWell/USA) The lower reaction mixture with mild shaking accompanied by room tempera-
absorbance indicated a higher free radical scaveng- ture incubation in a darkened position for 20 min-
ing function. The study revealed radical scaveng- utes (Geraghty et al., 2014).
ing behavior as the free radical inhibition factor.
This inhibition percentage measured by adopting RESULTS AND DISCUSSION
the coming formula,
% inhibition = (AC-AT) ÷ AC The synthesis of coumarinyl 1, 3, 4-oxadiazoles
from -3-carboxylic acid through several sequential
Ac: absorbance of the control (solvent + DPPH with-
steps (Figure 1) and FTIR con irmed their structures
out sample)
with 1 H NMR spectrometry. To achieve the desired
At: absorbance of the test sample. heterocycles, the sequence of reaction in the ig-
These tests were conducted in triplicates and the ure1 was followedCoumarin-3carboxylic acid ester-
indings were represented as an average value i ication with methanol in the existence of sulfuric
according (Sanja et al., 2009). acid resulted in coumarin3carboxylic acid methyl
ester (S1) characterized by the absence of a wide-
Anticoagulant activity band for OH stretching COOH group absorption in
A soft modi ication of a familiar procedure. Added coumarin-3 carboxylic acid and appearance of two
(0.1 ml) of 4 coumarin derivatives (S4 H, S4 F, S4 Cl, bands at 2933 cm-1 and 2880 cm-1 attributed to
S4 CO) with a series of concentrations (50, 100, C-H stretching vibration for CH3 group. The key
200) mg/ml, as optimistic control group, heparin intermediate for the synthesis of substituted one,
(1IU/µl), solvent as control group to (0.9 ml) plasma 3, 4-oxadiazole derivatives is coumarin-3-carboxylic
just incubated at 37 ◦ C for 10 minutes for the next acid hydrazide (S2), which was prepared by reac-
prothrombin time PT and enabled partial prothrom- tion of (S1) with hydrazine hydrate (80%). The
bin time APTT evaluation as directed by the man- FT-IR spectrum (S2) showed an absorption band in
ufacturer For PT calculation (0.2ml) of PT reagent the region of 3385 cm− 1 of the NH2 group and
spend 15 min prewarming at 37 ◦ C applied to the 3290 cm-1 of the NH group. The C = O stretching
plasma samples and control groups in the cuvette vibration was observed at 1614 cm-1 in the amid-
and calculated in seconds. Additionally, APPT group, respect to (S2) with various substituted ben-
assessment, the reagent was prewarmed at 37 ◦ C zaldehyde in moderate to good yield, to form C=N
for 2 minutes and applied to the samples, added the bond at 1687 cm−1 . Then oxidation of S3 deriva-
Ca-solution kit and recorded clotting times. We car- tives to give (S4 H, S4 Cl, S4 F, S4 OC) by acetyl anhy-
ried these steps out in triplicate for each samples dride to form coumarin -1,3,4-oxadiazole were con-
and control groups Raposo et al. (2015). irmed using FT-IR spectra showing C-O- C asym-
Anticancer activity metric and symmetric stretching bands at 1253 and
1199 cm-1 respectively. Therefore, the 1676 cm-1
Inconsiderable modi ication on approval procedure band for the C = N stretching combined with the dis-
It was accomplished by using the 96-well lat- bot- appearance of the NH2, NH and C=O amid stretch-
tomed microtitration plate, which involves three ing bands.Oxadiazole has an inductive effect due to
stages in this process. We separated the breast the presence of heteroatom in the ring and is known
cancer cell line from their lasks as they went to be a weak base. This consists of two pyridine-
via trypsinization to the subcon luent monolayer. like nitrogen that exhibits the character of the con-
Added to the falcons 20 ml of culture medium with jugated diene form (Bhat et al., 2005).
10% serum and combined with cells to prepare for
cell suspension It poured the suspension of cells into LD50 measurement
a culture lask a sterile beaker, then using microti- As summarized in (Table 1), the median lethal doses
tration, plate 96 well, 100 µl of cell suspension can were calculated for the newly synthesized coumarin
pass to each well using multi micropipette, plates derivatives (S4H, S4F, S4CL and S4Co), as following
protected with a disinfect adhesive ilm, lid put on, (1150, 450, 550 and 950) mg/kg, respectively. S4H

870 © International Journal of Research in Pharmaceutical Sciences

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Table 1: lethal dose evaluation of coumarin-oxadiazol derivatives

compound M0 M1 LD50 (mg/kg)
S4H 1100 1200 1150
S4F 400 500 450
S4Cl 500 600 550
S4CO 900 1000 950

M0= Highest calculated dose were no mortality observed.

M1= Lowest calculated dose were mortality observed.

and S4CO appear to be the safest compounds with

the highest LD50 values. On the other hand, S4F
and S4CL appear to be more toxic with the lowest
lethal dose. The high levels of LD50 for S4F and S4CL
appear to be similar to the results observed by Ghate
et al. with LD50 more than 1000mg/kg (Ghate et al.,
2005).
Antioxidant activity
Various compounds like polyphenols possess
impressive antioxidant with potent radical scav-
enger’s activity. In the present study, we use the
DPPH scavenging assay to evaluate the capability Figure 2: The effect of coumarin derivatives on
of the four synthesized coumarin derivatives as percent inhibition for IC50 assessments at
antioxidants. As shown in (Figure 2), free radical different concentrations. Ascorbic acid as +
scavenging activity was expressed as a percentage of control
radical scavenging (% inhibition). The more potent
antioxidant activity notice is the S4Cl compound
with IC50 1.49 followed by S4CO compound with
IC50 5.29, S4Fcompound with IC50 17.54, and S4H
compound with IC50 18.79. We used ascorbic acid
as a positive control for antioxidant comparison.
At low concentration, we had observed no signi i-
cant differences between all tested compounds and
standards. Increase dose associated with differ-
enced regarding S4F and S4CO, as shown in (Fig-
ure 3).
Actually, all newly synthesized coumarin deriva-
tives expressed considerable radical scavenging or Figure 3: The effect of coumarin derivatives on
antioxidant activity with IC50 range (1.49- 18.79) free radical scavenging activity at different
µg/ml. Our inding came in agreement with concentrations (% inhibition) associated with
Kenchappa et al. they found that the presence group comparison. P<0.05 considered
of electron-withdrawing functional groups have signi icantly different.
promising antioxidant activities (Kenchappa et al.,
2017).
Anticoagulant activity
In general heterocyclic coumarin, molecules have
been reviewed by Al-Majedy Y et al. for their radical Clinical research suggest that anticoagulants are
scavenging activity, a series of coumarin derivatives the top choices for thrombosis disorder prevention
with different substitutions gave rise to signi icant and treatment (Rishavy et al., 2018) in the present
free radical scavenging activity making synthesized research, we utilized PT and APTT to check the
derivatives promising molecules to act as antioxi- direct coagulation effect of coumarin derivatives. As
dants, anticancer and reducing cardiovascular dis- shown in (Figure 4) with respect to S4H, 200 mg/ml
eases with bene icial role in general health (Al- prolonged coagulation time in the APTT test signif-
Majedy et al., 2016). icantly compared to the control group but remain

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 Sanaryh Mohammed Al-Awad et al., Int. J. Res. Pharm. Sci., 2020, 11(1), 865-874

Figure 4: The effect of S4H on prothrombin time Figure 7: The effect of S4CL on prothrombin
and partial thromboplastin time in different time and partial thromboplastin time in
concentrations. Different characters (*, **, ***, different concentrations. Different characters
̸=) represent signi icantly difference between (*, **) represent signi icantly difference
groups in the same test P<0.05 between groups in the same test P<0.05

Figure 5: The effect of S4F on prothrombin time

and active partial thromboplastin time in Figure 8: The effect of coumarin derivatives on
different concentrations. Different characters percent inhibition for IC50 measurements at
(*, **, ***, ̸=) represent signi icantly difference different concentrations
between groups in the same test P<0.05

Figure 9: The effect of coumarin derivatives at

Figure 6: The effect of S4CL on prothrombin the different concentrations on the availability
time and partial thromboplastin time in of breast cancer cell ( % inhibition) associated
different concentrations. Different characters with group comparison. P <0.05 considered
(*, **) represent signi icantly difference signi icantly difference
between groups in the same test P<0.05

872 © International Journal of Research in Pharmaceutical Sciences

 Sanaryh Mohammed Al-Awad et al., Int. J. Res. Pharm. Sci., 2020, 11(1), 865-874

less than the positive control (heparin group) indi- agent (Kumar et al., 2012; Terzioglu and A, 2003).
cating anticoagulant activity. Whilst at low concen-
trations we watch no signi icant effect compared CONCLUSIONS
to control. To complete the idea about the coag-
ulation effect, we measured PT. The clotting time The study included the synthesis of the new
prolongs with increase dose, such prolongation less coumarin -1, 3, 4-oxadiazole compounds (S4H, S4F,
than that watch with heparin treated group. We S4Cl, and S4CO). This combination has been shown
realize a signi icant prolongation of clotting time to be an extremely useful tool for the development
with a high dose of S4F coumarin derivative com- of certain bioactive compounds., Taking into account
pared to control. An effect seems to be like with that derived from 3-carboxylic acid coumarin(S0)
heparin in both PT and APTT tests. Low doses and followed by the synthesis series of intermediate,
of S4F also prolong clotting time in the PT test S1, S2, S3H, S3F, S3CL, S3CO. The synthesized com-
only. As shown in (Figure 5). Proposition S4Cl pounds S4CL and S4CO were tested as anti-cancer
has showed in (Figure 6) we observed no signi i- agents against MCF7 breast cancer cell, which gave
cant prolongation of clotting times on low and high a signi icant activity, S4F and S4CO gave good activ-
doses on PT, except 200mg/kg slightly prolong on ity against clotting factor (as anticoagulant agents)
the APTT test. Regarding S4CO, signi icant prolon- PT and APTT comparing with heparin, and S4CL,
gation of both prothrombin time and APTT in a dose- S4CO and S4H which gave signi icant action against
dependent manner, as shown in (Figure 7) Anti- DPPH(as antioxidant agents). A good system for
coagulant studies have been conducted to improve developing and synthesizing recent and more pow-
prothrombin time (PT and partial thromboplastin erful drugs is the structure of such compounds.
active time (APTT) at a different tested does. Guru-
padayya and Balasubramanyam (2015) reported ACKNOWLEDGEMENT
that the presence of electron with drawl substitu-
tion increase the activity against coagulation factor The authors grateful the University of Basra, college
and coumarin derivatives presently available in the of pharmacy, department of pharmaceutical chem-
clinical ield have been the core of anticoagulation istry and department of pharmacology and toxicol-
therapy (Levine et al., 2004). ogy, private Bayan Group for Advanced Lab. diag-
nostics for their supporting to this work.
Anticancer activity
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