Synthesis, Characterization and Antioxidant Activity of some New
Antipyrine derived Schiff Bases
Mahwish Fatima1, Bharath Rathna Kumar. P1*, Venu Priya. R1, Sunil Kumar Kadiri2
1Department of Pharmaceutical Chemistry, Anwarul Uloom College of Pharmacy, Telangana, India.
2Department of Pharmacology, College of Pharmaceutical Sciences,
Dayanand Sagar University, Karnataka, India.
*Corresponding Author E-mail: bharathpharm@gmail.com
ABSTRACT:
Synthesis, characterization of a series of some new antipyrine derived Schiff bases 5(a-f) have been carried out and evaluated for their antioxidant activity. The newly synthesized compounds were characterized by physical and spectral data. The compounds 5(a-f) were measured for physical parameters and enlisted. The completion of the reaction, the progress of the reaction, and the purity of all newly produced compounds were monitored by TLC. Compounds in series 5(a-f) have yields that vary from 70 to 93%. Based on a review of the literature, there were no antipyrine derivatived Schiff bases reported till now, hence they were synthesized and characterized in this work. It was found that these moieties were useful in the field of pharmaceutical chemistry and that they demonstrated a variety of pharmacological actions. In the current work, six synthetic compounds 5(a-f) were prepared, and characterized by FTIR, 1HNMR and Mass spectroscopy studies. Melting points were determined and TLC was used to verify the purity of each compound. In an effort to produce new therapeutic compounds these compounds are tested for antioxidant properties and reported.
Schiff bases also known as azomethines or imines, recognised as intriguing scaffolds attracted a lot of researchers to synthesize various compounds possessing diverse pharmacological activities1. Schiff bases are often used in manufacture of pigments, corrosion inhibitors2, fluorescence substances3, catalysts, intermediates, metal complexes4,5. The pervasiveness of Schiff bases is due to their easiness of synthesis; hence a large number of compounds can be prepared in high yields by condensing various aromatic amines with carbonyl compounds. They are also synthesized by other advanced methods such as solvent free synthesis, microwave synthesis, and using catalysts like P2O5/Al2O3, ZnCl2, Lewis and Bronsted-Lowry acids etc.6
Schiff bases are versatile pharmacophores7 reported to exhibit various biological properties like analgesic8, antileishmanial9, antioxidant10, anti-inflammatory11,12, antiviral13, antifungal14 antibacterial15,16, and anticonvulsant activity17, anticancer activity18. It indicates that the presence of azomethine pharmacophore is essential for exhibiting these pharmacological activities. The synthesis of novel derivatives of antipyrine derived cobalt and copper complexes exhibiting antiparacitic activity instigated the interest in developing new antipyrine derived Schiff bases19. The pyrazolones are the most ancient synthetic drugs. Knorr was the first to prepare antipyrine, a drug with a pyrazolone structure, as a substitute for quinine20.
MATERIALS AND METHODS:
Benzene, chloroform, 4-hydroxy aniline, 4-methoxy aniline, 2-nitro aniline, phenyl hydrazine and ethyl acetoacetate were purchased from SD Fine chemicals, Mumbai, India. Inova 400 MHz NMR spectrometer, VG Autospec MS, were used to record NMR spectra and Mass spectra respectively.
|
S. No |
Compound |
R |
|
1 |
5a |
-H |
|
2 |
5b |
-4-OH |
|
3 |
5c |
4-OCH3 |
|
4 |
5d |
3-NH2 |
|
5 |
5e |
3-NO2 |
|
6 |
5f |
2-CH3 |
Scheme of synthesis
Procedure for the synthesis of 2,3-dimethyl-1-phenyl-pyrazole-5-one (antipyrine)21:
A mixture of 2ml of Ethyl acetoacetate and 1.5ml of phenyl Hydrazine was refluxed for 4hrs at 110-120℃ in an oil bath. The mixture was cooled and solidified by adding 4ml of ether and stirred vigorously. The crude product was recrystallized from ethanol. Dissolve the above product in mixture of 1ml Methanol and NaOH solution (230mg in 1ml water). To this add 0.66ml of dimethyl sulphate drop wise with continuous stirring. After complete addition reflux it on boiling water bath 1 hour. Then evaporate methanol add hot water to solution, filter the solution and extract the filtrate with benzene. Evaporate the benzene and crystallize the product with hot water or benzene.
General procedure for the synthesis of N-[(3Z)-1,5-dimethyl-2-phenyl-1,2-dihydro-3H-pyrazol-3-ylidene] aniline22 (5a-f):
In a 250ml clean flat-bottomed flask ethanolic solution of antipyrine (0.01mol) and aniline (0.01mol) was boiled under reflux for 6hrs.The solvent was then reduced to one-third of its volume and the resulting solution was cooled to 0oC.The solid product formed was removed by filtration and recrystallized from ethanol. The mobile phase used for performing TLC studies was benzene and chloroform in the ratio (8:2) v/v. The percentage yield of the compounds was found in the range of 70-93%.
Anti-Oxidant activity Procedures23:
General procedure for preparation of sample solutions:
Accurately weigh 100mg of sample and transfer in a 100ml volumetric flask and dissolved by using methanol. From the above stock solution 20ml was pipette out into 100ml volumetric flask. The final volume was made up to 100ml by transferring distilled water to obtain a concentration of 200μg/ml.
General procedure for preparation of standard solution:
Accurately weighed and transferred 100mg of Ascorbic acid into 100ml volumetric flask, dissolved by using little quantity of methanol and made up to the mark by using methanol. From the above stock solution 20ml was pipette out into 100ml volumetric flask. The final volume was made up to 100ml by adding distilled water to obtain the concentration of 200μg/ml.
Procedure for preparation of Phosphate Buffer:
6.8g of KH2PO4 and 1.56g of NaOH in 900ml distilled water was added and the pH was adjusted at 7.4 with NaOH solution and dilute with water to produce 1000ml.
Hydrogen peroxide scavenging activity:
Hydrogen peroxide causes lipid peroxidation and DNA mutations by converting into hydroxyl radicals. The absorption of H2O2 can be measured at 230 nanometers. The decrease in absorption of UV light due to test and standard compounds can be used for the quantification of hydrogen peroxide scavenging activity. A solution of 40 millimolar hydrogen peroxide was used as diluents for preparation of test and standard solutions in different concentrations (10-320μg/ml) were prepared by using phosphate buffer of pH 7.4. The free radical scavenging activity of both the ascorbic acid and sample solutions were determined at 200μg/ml concentration and mentioned in the table 10. About 3.4ml of each sample solution and standard solution were added to 0.6ml of hydrogen peroxide solution in separate test uses incubated for 40min at 30°C and the absorbance of the resulting solutions were measured by using UV spectroscopy at 230nm. Phosphate buffer was used as blank and a solution of 40 millimolar hydrogen peroxide was used as control for the study. The percentage scavenging activity of hydrogen peroxide was calculated by using the below formula
I % = [(Ac-As)/Ac] Χ 100 ... (1)
Where, Ac and As are absorbance of the control and test or standard respectively
Procedure for preparation of 40mM H2O2 solution:
1.140ml of H2O2 is dissolved in 250ml of phosphate buffer (pH7.4). The molarity of 30% H2O2 solution is 9.8M. To make 40mM solution, 30% H2O2 solution is diluted 245 times, i.e. mixing of 1mL of 30% H2O2 with 244ml of H2O will results in 245mL of 40mM H2O2.
DPPH Free Radical scavenging activity:
In vitro free radical scavenging activity of synthetic compounds was tested by using DPPH24. The standard used was ascorbic acid. The percentage inhibition was computed and contrasted with the reference value. The outcomes are displayed in Table No. 2.
RESULTS:
Phenyl Hydrazine on reaction with Ethyl acetoacetate in presence of ethanol undergoes cyclization to yield compound (3), which on further reaction with dimethyl sulphate to form antipyrine (4). Antipyrine was refluxed with several aromatic amines in presence of ethanol for about 6hrs to form the title compounds 5a-f. The physicochemical parameters of samples 5a-f were enlisted in table 1.
Spectral Data of Compounds 5a-f:
Spectral data of compound 5a:
IR-spectra: (V max cm-1): 3128(ArH), 2918, 2849 (CH3), 1597 (C=N str.), 1494 (N-CH3), 1302 (C-N str.), 1HNMR δppm 300 MHz, CDCl3: δ2.4 (s, CH3, 3H), δ3.3 (s, CH3, 3H), δ4.4 (s, CH, 1H), δ6.4-8.0(m, ArH, 10H), MS-m/z 263 (M+).
Spectral data of compound 5b:
IR-spectra: (V max cm-1): 3334 (OH str.), 2921, 2850 (CH3), 1594 (C=N str.), 1495 (N-CH3), 1309 (C-N str.), 1HNMR δppm 300 MHz, CDCl3: δ2.3 (s, CH3, 3H), δ3.1 (s, CH3, 3H), δ4.4 (s, CH, 1H), δ6.1-8.0(m, ArH, 9H), MS-m/z 279 (M+).
Spectral data of compound 5c:
IR-spectra: (V max cm-1): 3127(ArH), 2946, 2888 (CH3), 1597 (C=N str.), 1495 (N-CH3), 1302 (C-N str.), 1HNMR δppm 300 MHz, CDCl3: δ2.3 (s, CH3, 3H), δ3.2 (s, CH3, 3H), δ3.9 (s, OCH3, 3H), δ4.3 (s, CH, 1H), δ6.2-8.0(m, ArH, 9H), MS-m/z 294 (M+1).
Spectral data of compound 5d:
IR-spectra: (V max cm-1): 3348 (NH2 str.), 2952, 2849 (CH3), 1593 (C=N str.), 1492 (N-CH3), 1312 (C-N str.), 1HNMR δppm 300 MHz, CDCl3: δ2.3 (s, CH3, 3H), δ3.3 (s, CH3, 3H), δ4.3 (s, CH, 1H), δ5.3 (s, amine NH2, 2H,) δ6.4-8.0(m, ArH, 9H), MS-m/z 278 (M+).
Spectral data of compound 5e:
IR-spectra: (V max cm-1): 3126(ArH), 2953, 2850 (CH3), 1597 (C=N str.), 1495 (N-CH3), 1301 (C-N str.), 1HNMR δppm 300 MHz, CDCl3: δ2.4 (s, CH3, 3H), δ3.3 (s, CH3, 3H), δ4.2 (s, CH, 1H), δ6.4-8.0(m, ArH, 10H), MS-m/z 309 (M+1).
Spectral data of compound 5f:
IR-spectra: (V max cm-1): 3125(ArH), 2902, 2834 (CH3), 1594 (C=N str.), 1497 (N-CH3), 1313 (C-N str.), 1HNMR δppm 300 MHz, CDCl3: δ2.1 (s, CH3, 3H), δ2.4 (s, CH3, 3H), δ3.3 (s, CH3, 3H), δ4.2 (s, CH, 1H), δ6.2-8.0(m, ArH, 9H), MS-m/z 277 (M+).
Table 1: Physical properties of Antipyrine derivatives (5a-f)
|
Sample Code |
Chemical formula |
M.Wt (g/mol) |
M.P (oC ) |
Percentage yield |
TLC Retardation factor value |
|
5a |
C17H17N3 |
263.3 |
117-118 |
86 |
0.54 |
|
5b |
C17H17N3O |
279.3 |
139-140 |
75 |
0.46 |
|
5c |
C18H19N3O |
293.3 |
155-156 |
82 |
0.74 |
|
5d |
C17H18N4 |
278.3 |
162-163 |
70 |
0.62 |
|
5e |
C17H17N4O2 |
308.3 |
110-111 |
93 |
0.68 |
|
5f |
C18H19N3 |
277.3 |
114-115 |
72 |
0.52 |
DISCUSSION:
In the present study synthesis and characterization of some new antipyrine derivatives 5a-f was carried out and the results are presented here. The structures of the compounds were elucidated by physical and spectroscopic data the physical characteristics such as melting point, retardation factor (Rf), molecular weight, molecular formula and percentage yield were recorded and enlisted in the table 5. The purity of the compounds 5a-f was continuously monitored by using thin layer chromatography. The percentage yield of the compounds 5a-f synthesized was ranging from 70-93%. The IR spectra of compound 5a exhibited characteristic absorption bands at 3128 due to aromatic C-H stretching, 2918 and 2849 due to aliphatic C-H stretching, 1597 due to C=N stretching. The 1HNMR exhibited a singlet peak at δ2.4, due to NCH3 protons, peak at δ3.3, due to CCH3 protons, peak at δ4.3, due to CH proton of pyrazole ring and multiplet at δ6.4-8.0 due to 10 aromatic protons. Mass spectra showed molecular ion peak at 263m/z value which confirms the formation of compound 3a. The results are mention in the table 2.
Table 2: Antioxidant Activity data of Compounds 5a-f.
|
Compounds |
Scavenging activity (in %) on |
|
|
Hydrogen peroxide |
DPPH radical |
|
|
5a |
30.50 ± 2.50 |
35.96±1.58 |
|
5b |
83.05 ± 2.09 |
76.38±3.25 |
|
5c |
66.94 ± 2.86 |
56.16±2.68 |
|
5d |
61.86 ± 1.95 |
52.46±3.17 |
|
5e |
52.54 ±4.70 |
45.38±2.43 |
|
5f |
23.73 ± 2.75 |
22.19±2.32 |
|
Ascorbic acid (Std.) |
96.75± 1.35 |
94.25±2.80 |
CONCLUSION:
The present research work about synthesis and characterization of some new antipyrine derivative Schiff bases have been carried out and tested for antioxidant activity. Out of all the synthesized compounds, 5b compound is showing good antioxidant activity and compound 5a is showing least antioxidant activity was reported in this study. The compounds 5b and 5c showed good antioxidant activity due to phenolic OH group and electron releasing OCH3 groups. These two compounds can be used as lead compounds for designing potent antioxidant compounds.
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Received on 07.02.2024 Modified on 11.03.2024
Accepted on 03.04.2024 ©AJRC All right reserved
Asian J. Research Chem. 2024; 17(2):73-77.
DOI: 10.52711/0974-4150.2024.00014