Synthesis and evaluation of novel Flavonoid derivatives for Antibacterial activity
Sushil D. Patil1*, Masood Ahemed Hafizur
H1, Aher Priti
R.1, Pravin B. Shelke1,
Samruddhi Yardi2
1Department of Pharmaceutical Chemistry, MET
Institute of Pharmacy, Adagaon, Nashik,
S.P. Pune University, M.S., India.
2T.Y.B. Pharm, Department of Pharmaceutical Chemistry, MET Institute
of Pharmacy, Adagaon, Nashik,
S.P. Pune University, M.S., India.
*Corresponding Author E-mail:
ABSTRACT:
Flavones
are the derivatives of flavonoids, possessing the
wider range of biological actions. A series of flavonoid
derivatives with the help of substituted benzaldehyde
were synthesized and evaluated for antibacterial activity by cup-plate agar
diffusion method. Out of six compounds 1-(2, 4- dihydroxyphenyl)-3-(4-(dimethyl amino) phenyl) prop-2ene-1one.(MCR-001)
and 2-(4-(Di-methylamino)
phenyl)-7-hydroxy-4-H-chromen-4-one. (MCR-002),
2-hydroxy-2-(4-(methyl (4-nitrostyryl) amino) phenyl)-4-oxochroman-7-yl
benzoate. (MCR-004) and 2-(4-dimethyl amino)
phenyl)-4-oxo-4H-chromen-7-yl benzoate. (MCR-005) 2-(4-dimethyl amino)
phenyl)-2-hydroxy 4-oxochroman-6-yl benzoate (MCR-005) have shown promising
antibacterial activity also 7-hydroxy-2-(4-methyl (4-nitrostyryl) amino)
phenyl)-4-H-chromen-4-one (MCR-003) ) and 2-(4-(4-chlorobenzyl) methyl) amino)
phenyl)-2, 6-dihydroxychroman-4-one. (MCR-006) shown moderate
antibacterial activity. This work has revealed the scope and potential
of flavonoids for future exploration to synthesize
substituted flavonoids and chalcone
derivatives by the use of different substituted benzaldehyde.
KEY WORDS: Flavonoid, agar diffusion method, antibacterial activity.
INTRODUCTION:
Flavonoid
has gained recent interest because of their broad biological and
pharmacological activities including anti‐oxidant
[1‐4], Anxiolytic[5], anti cancer[6], analgesic[7‐8], and antimicrobial[9] activities. Flavonoid showed promise of being powerful antioxidants
which can protect the human body from free radicals. The function of an
antioxidant is to intercept and react with free radicals at a rate faster than
the substrate.
Since free radicals are able to attack at
a variety of target including lipids, fats and proteins, it is believed that
they may damage organisms, leading to disease, poisoning and including aging[10]. These derivatives as ref.[11]
synthesized. The chemical structures of the new derivatives were confirmed by
elemental and spectral (1H-NMR and Mass) analyses. All compounds were
investigated for antibacterial activity
MATERIALS
AND METHODS:
All materials were procured from Sigma
Aldrich and Merck specialties Pvt. Ltd. (Mumbai. India). Solvents were dried
and distilled being used. Anhydrous sodium sulphate was used to dry the
solvents.
Figure 1
Thin Layer Chromatography (TLC) analyses
were carried out on aluminum plates (Merck) precoated
with silica gel 60 F254 (0.2 mm), and spots were visualized with UV light and
I2. Liquid intermediates were checked for purity using Gas chromatography (Pack
column SE-30, OV-101, and capillary column BP-5). Melting points were taken in
open glass capillary using OMEGA melting point apparatus and were uncorrected.
Infra red (IR) spectra were recorded on KBr pellets
on a Shimadzu 1000 FTIR spectrometer in the range of 4000-200 cm-1, Resolution
2.0 with number of scan - 45. Apodization; Happ-Genzel. Proton (1H) Nuclear
Magnetic Resonance (NMR) spectra of compounds were recorded on Bruker Advance II 400 NMR Spectrophotometer using CDCl3
solvent, at SAIF, Punjab University, Chandigarh. The method followed for the
synthesis of flavonoids derivatives has been
elaborated here:
Synthesis:
All these derivatives 1-(2,4- dihydroxyphenl)-3-(4-(dimethyl
amino)phenyl) prop-2ene-1one.(MCR-001) ,2-(4-(Dimethylamino)phenyl)-7-hydroxy-4-H-chromen-4-one.
(MCR-002), 7-hydroxy-2-(4-methyl
(4-nitrostyryl) amino)phenyl)-4-H-chromen-4-one.
(MCR-003), 2-hydroxy-2-(4-(methyl(4-nitrostyryl)amino)phenyl)-4-oxochroman-7-yl
benzoate. (MCR-004), 2-(4-dimethyl
amino)phenyl)-4-oxo-4H-chromen-7-yl benzoate.
(MCR-005)and
2-(4-(4-chlorobenzyl) methyl) amino)
phenyl)-2, 6-dihydroxychroman-4-one. (MCR-006) are synthesized as mention in [11]
Synthesis
of 1-(2,4-
dihydroxyphenl)-3-(4-(dimethyl
amino)phenyl) prop-2ene-1one.(MCR-001 )
IR: 3059, 2916 (Ar-CH), 2821 (-CH), 1681(Ar-C=C), 1778 (-C=O), 3350 (-OH)
1H-NMR: 7.3-8.5(Doublet (2H) Benzene -CH ), 5.35 (Singlet Ar-OH ), 7.4
(Doublet Ar-CH) 3.09( Singlet (6H) CH3).
Synthesis of 2-(4-(Dimethylamino)phenyl)-7-hydroxy-4-H-chromen-4-one(MCR-002)
IR: 2900, 3049, 2819(Ar-CH),
729, 825 Benzene (p), 1678(-C=O), 3317(-OH)
1H-NMR: 6.69-7.18 (Doublet (2H)Ar -CH), 5.40 (Singlet Ar-OH), 3.08 (Singlet (6H) CH3),
7.5-7.6( Doublet –CH)
Table
no 1
|
Sr.
no. |
Molecular
Formula |
Mol.Wt. gm/mol |
%
Yield |
M.P.
(0C) |
Mobile Phase |
Rf value. |
|
1 |
C17H17NO3 |
283.32 |
73% |
48o-50oC |
nH : EtA 7:5 |
0.8 |
|
2 |
C17H15NO3 |
299.32 |
62% |
62º-65ºC |
nH : EtA 7:5 |
0.698 |
|
3 |
C24H18N2O5 |
420.41 |
58% |
58º-60ºC |
nH : EtA 5:5 |
0.733 |
|
4 |
C31H24N2O7 |
542.52 |
55% |
80º-82ºC |
Ben :meth 7:3 |
0.615 |
|
5 |
C24H19NO4 |
393.43 |
53% |
90º-93ºC |
Ben : meth 7:3 |
0.646 |
|
6 |
C23H20ClNO4 |
409.86 |
55% |
75º-78ºC |
nH : EtA 5:5 |
0.600 |
EtA: Ethyl acetate, nH:
n-Hexane, Ben : Benzene, meth: methanol.
Synthesis
of 7-hydroxy-2-(4-methyl
(4-nitrostyryl) amino) phenyl)-4-H-chromen-4-one (MCR-003)
IR: 3080 (Ar
-C-H), 1446, 1539(NO2), 1705 (C=O), 3309 (Ar-OH),
1670 (Ar-C=C)
1H-NMR: 6.69-8.40(Doublet Ar-CH), 5.69 (Singlet Ar-OH),
3.08 (Singlet (6H) CH3), 6.78(Doublet-CH)
Synthesis
of 2-hydroxy-2-(4-(methyl
(4-nitrostyryl) amino) phenyl)-4-oxochroman-7-yl benzoate (MCR-004)
IR: 2825, 2991, 3072 (Ar
-C-H), 1166, 1294(C-N), 1496, 1531, 1454 (NO2), 1712, 1703 (C=O),
1598 (Ar-C=C) 711, 815 (Benzene), 1788(-COOR) 1H-NMR:
6.69-8.1 (Doublet Ar-CH), 5.69 (Singlet Ar-OH), 3.09 (Singlet (6H) CH3),
Synthesis
of 2-(4-dimethyl amino) phenyl)-4-oxo-4H-chromen-7-yl
benzoate (MCR-005)
IR: 2823, 3030, 3070 (Ar
-C-H), 1708 (C=O), 1670 (Ar-C=C), 812-707( Benzene), 1166,1128 (C-N), 1791(-COOR) 1H-NMR: 7.1-8.2
(Doublet(6H)Ar-CH), 5.38 (Singlet Ar-OH),
3.1 (Singlet (6H) CH3)
Pharmacological
evaluation
Antibacterial
activity
The compounds
were tested in-vitro for their antibacterial activity against two
microorganisms viz. Escherichia coli (NCTC 10418), and Staphylococcus
aureus (NCTC 6571) which are pathogenic in human
beings.
a)
Method:
Cup-plate agar diffusion method using Nutrient agar.
b)
Preparation of test solutions:
Each test compound (5 mg) was dissolved in dimethyl formamide (5 mL) to give stock solution of concentration 100 mcg/mL. Then 0.1 mL of this solution
was used for testing.
c)
Preparation of standard solution:
Standard drug Norfloxacin
was used. The concentration was 100 mcg/mL.
d)
Method of testing:
Nutrient agar plates were prepared by pouring 15-20 mL of the medium into each sterilized petridish
and were allowed to set at room temperature. The cell suspension was
standardized to the density of 530 nm using spectrophotometer and was
inoculated over the surface of agar medium using sterile cotton swab. The three
cups were scooped in each plate using a sterile cork borer of 6 mm diameter.
Then the solutions of test compounds (0.10 mL) were
added in cups by using micropipettes and these plates were incubated at 37°C for 48 hrs. The zone of inhibition was measured in mm for each
organism.
Table No.2 Antibacterial activities of the synthesized compounds
|
SL. NO. |
Compd. |
Zone
of inhibition at 100 mcg/mL (in
mm.) |
|
|
E. coli |
S. aureus |
||
|
1 |
(MCR-001) |
23 |
23 |
|
2 |
(MCR-002) |
23 |
22 |
|
3 |
(MCR-003) |
18 |
19 |
|
4 |
(MCR-004) |
23 |
24 |
|
5 |
(MCR-005) |
24 |
23 |
|
6 |
(MCR-006) |
17 |
16 |
|
Standard |
Norfloxacin |
23 |
24 |
CONCULSION:
A series of flavonoid derivatives with the help
of substituted benzaldehyde were synthesized and
evaluated for antibacterial activity by cup-plate agar diffusion method. Out of six compounds 1-(2, 4- dihydroxyphenyl)-3-(4-(dimethyl amino) phenyl) prop-2ene-1one. (MCR-001) and 2-(4-(Di-methylamino) phenyl)-7-hydroxy-4-H-chromen-4-one.
(MCR-002), 2-hydroxy-2-(4-(methyl (4-nitrostyryl)
amino) phenyl)-4-oxochroman-7-yl benzoate. (MCR-004) and 2-(4-dimethyl amino)
phenyl)-4-oxo-4H-chromen-7-yl benzoate. (MCR-005)
2-(4-dimethyl amino) phenyl)-2-hydroxy 4-oxochroman-6-yl benzoate (MCR-005)
have shown promising antibacterial activity also 7-hydroxy-2-(4-methyl
(4-nitrostyryl) amino) phenyl)-4-H-chromen-4-one (MCR-003) ) and
2-(4-(4-chlorobenzyl) methyl) amino) phenyl)-2, 6-dihydroxychroman-4-one.
(MCR-006) shown moderate antibacterial activity. Norfloxacin was used as standard drug. The promising biological activities of these
compounds are taken into consideration for drug development and drug discovery
study.
ACKNOWLEDGEMENTS:
We are very thankful to the Management of
MET’s Institute of Pharmacy for providing infrastructural facilities to carry
out the research work. Also we are very thankful to SAIF, Punjab University for
providing analytical instrumentation Facility.
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Received on 10.12.2015 Accepted
on 01.01.2016
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Asian J. Pharm. Res. 6 (1): January -March, 2016; Page 27-30
DOI: 10.5958/2231-5691.2016.00005.8