Graph theoretical analysis
Proteins (nodes) and interactions (edges) were re-created as a graph, which was depicted in Figure 2 and Table 1. The network has 59 nodes and 82 edges, and the characteristics were used to determine the relevance of proteins. CXCR6/Chemokine, FGR, PIK3CA, PTK2, GNB5, CDC42, AKT3, HRAS, RAC1, PIK3R6, PLCB1, GNAI1, VAV3, ITK, PRKCB, PRKCZ, RASGRP2, PTK2B, and PREX1 are high significant proteins based on the average measure of each parameter. CXCR6/Chemokine had a Centrality score of 2262 for Stress, a score of 9 for Radiality, a score of 0.439326 for Eigenvector, a score of 0.125 for Eccentricity, a score of 0.007299 for Closeness, a score of 944.0443 for Betweenness, and a score of 12 for Degree from the network among the best 19 nodes. Based on significant metrics with its threshold values, the target CXCR6/Chemokine was identified as a therapeutic target for the treatment of inflammation. Because of its interaction with 19 main proteins that are involved in inflammation, CXCR6/Chemokine has gotten increasing attention. CXCR6/Chemokine was picked as a noteworthy target based on the graph theoretical analysis report and importance.
Druglikeness analysis
The activity scores for the drug relative to "average drug-like molecules" are displayed in the Figure 3. The greater the value of the score, the more likely it is that the particular molecule will be active. The analysis report was conforming that the chosen 5a-5l test compounds were having kinase protein inhibitory activity.
Insilico Modelling
Molecular docking studies of was carried out with the synthesized 3-(4-(5-(substituted phenyl)-3-phenyl-4,5-dihydro-1H-pyrazole-1-carbonyl) phenyl imino)-1-(substitutedmethyl)indolin-2-one (5a-5l) test substance, as well as the standards Acetaminophen, Celecoxib, Diclofenac, Indomethacin, Naproxen, Ibuprofen against CXCR6 (PDB: 6KVA). Among tested ligands 5a, 5c, and standardCelecoxib showed high amino acid interaction and significant binding affinity towards CXCR6 protein with the biding energy of -11.4, -11.0 and -8.8 Kcal/mol respectively. The amino acids interaction with the 5a was ARG39[2.64 Å], ARG86[3.06 Å, 4.71Å], LEU87[3.14 Å], ARG88[3.96 Å], ASP90[4.05 Å], GLNP111[3.13 Å,11.72Å], VAL154[2.63 Å],THR155[3.68 Å],VAL156[2.10 Å]. The amino acids interaction with the 5b was GLN1[3.12 Å], PRO46[3.15 Å], PHE106 [16.51 Å,3.15Å], GLY108[3.52 Å], GLN179[4.12 Å], SER180[3.78 Å], SER185[3.19 Å]. The amino acids interaction with the 5c was GLN1[2.00 Å, 3.31 Å, 3.31 Å, 4.53 Å, 15.37 Å], PRO42[3.94 Å], GLU47[3.55 Å], LYS64[3.59 Å], TRP104[3.32 Å], VAL105[3.14 Å],PHE106[2.16 Å]. The amino acids interaction with the 5d was GLN6[3.59 Å], ARG39[5.17 Å], ALA41[2.87 Å], GLY45[, 2.24 Å 2.97 Å ], GLU47[3.19 Å , 3.84 Å], GLY108[3.33 Å]. The amino acids interaction with the 5e was GLN44[2.39 Å, 2.39 Å, 3.51 Å], ASP90[4.75 Å], VAL154[3.88 Å], VAL169[3.84 Å], THR171[11.12 Å]. The amino acids interaction with the 5f was ARG88[5.12 Å], PRO89[5.00 Å], ASP90[4.26 Å], GLY152[2.87 Å], ALA153[3.51 Å], THR171[3.80 Å]. The amino acids interaction with the 5g was GLN1 [2.05 Å, 3.69 Å], GLU47 [3.72 Å], LYS64 [4.59 Å, 5.06 Å], TRP104[3.64 Å], THR173[3.50 Å], ALA176[3.04 Å]. The amino acids interaction with the 5h was GLN1[3.25 Å], ARG39[2.78 Å], GLN44[2.29 Å], GLY45[2.71 Å], GLU47[3.64 Å], ASP90[3.31 Å], GLP168[3.37 Å], SER180[2.67 Å]. The amino acids interaction with the 5i was GLP40[2.49 Å, 3.93 Å], GLY43[4.18 Å, 3.71 Å], GLN44[ 2.45 Å], ASP93[3.65 Å], TYR96[1.88 Å]. The amino acids interaction with the 5j was TRP158[1.26 Å], VAL165[1.93 Å], LYS166[3.67 Å], TYR184[5.48 Å]. The amino acids interaction with the 5k was GLN44 [2.36 Å, 3.24 Å], GLU47[3.49 Å], ASP90[4.70 Å], VAL154[3.85 Å, 3.00 Å], VAL169[3.80 Å], THR171[11.18 Å]. The amino acid interaction with the 5l was GLP40 [2.46 Å, 4.00 Å], GLY43[3.78 Å], GLN44[2.62 Å], ASP93[3.57 Å],VAL94[3.81 Å], TYR96[1.97 Å], LYS166[2.29 Å]. The amino acids interaction with the Celecoxib was GLN1[4.16 Å], LEU4[3.03 Å], GLU47[4.26 Å,4.26 Å], TRP48[2.25 Å], TRP104[1.95 Å], PHE106[3.04 Å, 3.80 Å, 2.99 Å], GLY108[2.89 Å], SER180[2.97 Å]. The amino acids interaction with the Aspirin was ARG34[4.83 Å, 2.97 Å], TRP99[3.06 Å].The amino acids interaction with the Acetaminophen was HSD100[2.21 Å], VAL119[2.88 Å], SER121[1.96 Å]. The amino acids interaction with the Diclofenac was ALA2[3.28 Å], PRO46[4.07 Å], TRP48[2.11 Å], GLN63[2.46 Å], TRP104[3.36 Å, 4.03 Å], VAL105[3.90 Å, 3.99 Å]. The amino acids interaction with the Ibuprofen was GLN1[2.39 Å], ARG25[15.81 Å], SER121[2.16 Å], ALA122[2.14 Å]. The amino acids interaction with the Indomethacin was GLN1[2.67 Å], PRO46[2.96 Å], GLU47[3.40 Å, 4.00 Å], PHE106[3.27 Å], GLY107[2.21 Å]. The amino acids interaction with the Naproxen was ARG25[15.76 Å], HSD100[3.29 Å], SER120[2.70 Å], SER121[2.27 Å], ALA122[2.25 Å]. The binding interactions, 2D and 3D model of 5a, 5c, and Celecoxib were shown in the Table 2 and Figure 4-6.
In vivo antiinflammatory activity
Antiinflammatory tests have been performed for compounds 5a-5l and standard Diclofenac Celecoxib by using groups of Swiss albino mice at the dose of 250 mg kg-1. All of the tested compounds were exhibited antiinflammatory activity compared with the standards. Both of these standards were commercially available and also it was used for relating purposes with dose level of 250 mg kg-1. Compound 5a showed the highest activity reducing the paw edema by 74.2%. Pyrimidines 5c and 5l presented 48.1% and 40.4% of inflammation reductions, respectively (Table 3, 4 and Figure 7).
Density functionality theory
The stability and bioactive nature of 5a, 5c, and Celecoxib was measured by using energy values of HOMO and LUMO. From the energy values of HOMO and LUMO, the energy gap was calculated to measure the stability and bioactivity of test compound, and standard. The test compound 5a HOMO energy value was 0.28634, LUMO energy value was 0.21216 and HOMO and LUMO energy gap was 2.018541652. Likewise the 5c HOMO and LUMO energy gap was 1.98776355 and Celecoxib HOMO and LUMO energy gap was 3.57253219. Based on the obtained data was suggesting that the test compounds was having significant stability as par with the standard. The energy values of HOMO and LUMO of test compound, and standard were shown in the Table 5 and Figure 8-10
Pharmacokinetic and physicochemical properties prediction analysis
The ADME and physicochemical properties of selected 5a, 5c, and Celecoxib were assessed through SwissADME (http://www.swissadme.ch/) webserver and these are presented in Table 6. From the assessed data in Table 6, the 5a, 5c, and Celecoxib was found not to violate Lipinski’s rule of five. The polar surface area of test compound 5a was 64.68 Ų, 5c was 90.70 Ų and Celecoxib 88.99Ų. The findings also showed that the 5a, 5c, and Celecoxib had high human gastro intestinal (GI) absorption, In general, increased GI absorption leads to increased chemical bioavailability. As a result, oral administration of the test substance 5a, 5c may result in improved absorption from the gastrointestinal system.
The higher numbers of H-bonds are possibly measured to be involved during protein ligand binding. From the result, the bioavailability score of three compounds showed better results +0.55 for 5a, 5c, and Celecoxib. Thus relating with molecular properties 5a, 5c, and Celecoxib was predicted to have better chances as a possible drug-relevant candidate with antiinflammatory potential.
The test compounds and standard are soluble in nature, the synthetic accessibility score was found to be >6 for 5a with 3.66, 5a with 3.57, Celecoxib with 3.70, which indicated that the all the compounds are very feasible to synthesize.
The graphical representations of lipinski rule of selected compounds are presented in the Figure 11. The pink area within the hexagon represents the optimal range of the compounds. The recommended range for drug-like compound was in saturation (INSATU): fraction of carbons in the sp3 hybridization not less than 0.25, insolubility (INSOLU): log S not higher than 6, hydrophobicity (LIPO): between -0.7 and +5.0, rotatable bonds (FLEXI): no more than 9 rotatable bonds, molecular weight (SIZE): between 150 and 500 g.mol-1, polar surface area (POLAR): The red slanted hexagon inside the pink tint represents the reference compounds, which have drug-like characteristics. Based on the observation 5a, 5c, and Celecoxib had drug-like qualities.
Furthermore, the pharmacokinetic parameters of the chosen chemical, and standard, were studied using the egg-boiled model represented in Figure 12. Predicting passive gastrointestinal absorption and BBB penetration using the egg-boiled model is useful because it takes into account two important pharmacokinetic features at once. Egg-shaped plot demonstrates that chemical in yolk (i.e. yellow area) represents very possible BBB permeability and albumin (i.e. white region) represents highly probable human intestine absorption, as shown by the egg-shaped plot of organisational structure. From the Figure 12, the 5a, 5c, and Celecoxib found in albumin (white region) elucidated the good absorption in gastrointestinal region. From the above observed results, it can be interpreted that the 5a and 5c compounds have sufficient potential to be drug.
Synthesis of 1-(substitutedmethyl)indoline-2,3-dione (2a-2b)
A mixture of isatin (1.47 g; 0.01 mol), formaldehyde (0.45 g; 0.015 mol), and morpholine/piperidine (0.87/0.85 g; 0.01 mol) in ethanol (25 ml) was stirred for 2 h in magnetic stirrer. Then the resulting mixture was refluxed on water bath for 4 h. The above mixture was poured on crushed ice and mixed well. The solid 2a/2b which obtained was filtered, dried, and recrystallised using rectified spirit.
1-(Morpholinomethyl)indoline-2,3-dione (2a)
Yield: 77 % (1.89 g), m.p.: 243-245 °C, Rf Value: 0.49 [Toluene: n-Propanol (40: 60)]. IR (KBr, cm‒1): 3094 (Ar-CH), 2937 (CH2-CH), 1692 (C=O), 1613 (C=C), 1040 (C-O-C). 1H NMR (CDCl3, 500 MHz) δ ppm: 2.13-4.35 (m, 8H, CH2 of morpholine), 4.96 (s, 2H, CH2 linkage), 7.28-7.90 (m, 4H, Ar-H). 13C-NMR (δ: ppm): 51.7 (C-2’ & C-6’), 63.2 (C-3’ & C-5’), 74.5 (CH2 linkage), 119.1 (C-8), 124.0 (C-7), 125.8 (C-5), 130.3 (C-4), 136.9 (C-6), 151.6 (C-9), 158.2 (C-2), 188.4 (C-3). EI-MS m/z: 246 (M+). Anal. Cald for C13H14N2O3: C, 63.40; H, 5.73; N, 11.38. Found: C, 63.62; H, 5.71; N, 11.35.
1-(Piperidin-1-ylmethyl)indoline-2,3-dione (2b)
Yield: 73 % (1.78 g), m.p.: 219-221 °C, Rf Value: 0.62 [Toluene: n-Propanol (40: 60)]. IR (KBr, cm‒1): 3069 (Ar-CH), 2922 (CH2-CH), 1687 (C=O), 1591 (C=C). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.87-2.99 (m, 10H, CH2 of piperidine), 4.53 (s, 2H, CH2 linkage), 7.02-7.97 (m, 4H, Ar-H). 13C-NMR (δ: ppm): 24.7 (C-3’ & C-5’), 25.1 (C-4’), 52.3 (C-2’ & C-6’), 72.8 (CH2 linkage), 118.0 (C-8), 123.3 (C-7), 125.2 (C-5), 128.6 (C-4), 133.5 (C-6), 148.9 (C-9), 159.3 (C-2), 185.0 (C-3). EI-MS m/z: 244 (M+). Anal. Cald for C14H16N2O2: C, 68.83; H, 6.60; N, 11.47. Found: C, 69.06; H, 6.61; N, 11.43.
Synthesis of 4-(1-(substituted methyl)-2-oxoindolin-3-ylideneamino) benzoic acid (3a-3b)
Equimolar quantity of 1-(substitutedmethyl)indoline-2,3-dione 2a/2b (2.46/2.44 g; 0.01 mol)and p-amino benzoic acid (1.37 g; 0.01 mol) in ethanol (30 ml) was refluxed on sand bath for 5 h. The reaction mixture was cooled to room temperature, poured in crushed ice and kept aside overnight in refrigerator. The product separated 3a/3b was filtered, dried and recrystallised using ethanol.
4-(1-(Morpholinomethyl)-2-oxoindolin-3-ylideneamino)benzoic acid (3a)
Yield: 70 % (2.56 g), m.p.: 142-144 °C, Rf Value: 0.35 [Toluene: n-Propanol (40: 60)]. IR (KBr, cm‒1): 3405 (OH), 3071 (Ar-CH), 2946 (CH2-CH), 1729 (C=O), 1668 (C=N), 1594 (C=C), 1072 (C-O-C). 1H NMR (CDCl3, 500 MHz) δ ppm: 2.01-4.14 (m, 8H, CH2 of morpholine), 4.48 (s, 2H, CH2 linkage), 6.95-8.22 (m, 8H, Ar-H), 10.09 (s, 1H, OH). 13C-NMR (δ: ppm): 50.8 (C-2’ & C-6’), 65.1 (C-3’ & C-5’), 72.0 (CH2 linkage), 119.4 (C-8), 122.5 (C-7), 123.0 (C-2” & C-6”), 124.7 (C-5), 127.6 (C-4”), 129.1 (C-4), 132.3 (C-6), 132.9 (C-3” & C-5”), 150.2 (C-9), 157.5 (C-1”), 162.9 (C-3), 163.4 (C-2), 168.6 (COOH). EI-MS m/z: 365 (M+). Anal. Cald for C20H19N3O4: C, 65.74; H, 5.24; N, 11.50. Found: C, 65.51; H, 5.26; N, 11.53.
4-(2-Oxo-1-(piperidin-1-ylmethyl)indolin-3-ylideneamino)benzoicacid (3b)
Yield: 78 % (2.83 g), m.p.: 130-132 °C, Rf Value: 0.48 [Toluene: n-Propanol (40: 60)]. IR (KBr, cm‒1): 3422 (OH), 3058 (Ar-CH), 2910 (CH2-CH), 1719 (C=O), 1681 (C=N), 1596 (C=C). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.95-3.27 (m, 10H, CH2 of piperidine), 4.32 (s, 2H, CH2 linkage), 7.11-8.38 (m, 8H, Ar-H), 9.94 (s, 1H, OH). 13C-NMR (δ: ppm): 26.0 (C-3’ & C-5’), 26.4 (C-4’), 52.7 (C-2’ & C-6’), 69.8 (CH2 linkage), 115.2 (C-8), 121.6 (C-7), 122.9 (C-2” & C-6”), 123.4 (C-5), 130.1 (C-4”), 130.8 (C-4), 132.6 (C-6), 133.0 (C-3” & C-5”), 149.5 (C-9), 156.2 (C-1”), 164.6 (C-3), 164.9 (C-2), 170.3 (COOH). EI-MS m/z: 363 (M+). Anal. Cald for C21H21N3O3: C, 69.41; H, 5.82; N, 11.56. Found: C, 69.62; H, 5.81; N, 11.54.
Synthesis of 4-(1-(substitutedmethyl)-2-oxoindolin-3-ylideneamino) benzohydrazide (4a-4b)
To 4-(1-(substitutedmethyl)-2-oxoindolin-3-ylideneamino)benzoicacid 3a/3b (3.65/3.63 g; 0.01 mol), thionyl chloride (1.78 g; 0.015 mol) was added slowly with stirring and refluxed gently for 2 h. The excess of thionyl chloride was distilled off, and the reaction mixture was cooled in ice bath. The product thus formed was immediately used for next step. To the above product, ethanol (50 ml) was added. To this solution 95 % hydrazine hydrate (1 g; 0.02 mol) was slowly added with stirring. Then the mixture was refluxed for 10 h followed by removal of excess solvent under reduced pressure, and resultant solution was poured in ice cold water. The product separated 4a/4b was filtered, and recrystallised from ethanol.
4-(1-(morpholinomethyl)-2-oxoindolin-3-ylideneamino)benzohydrazide (4a)
Yield: 72 % (2.73 g), m.p.: 166-168 °C, Rf Value: 0.76 [Hexane: Chloroform: n-Butanol (20: 30: 50)]. IR (KBr, cm‒1): 3347 & 3283 (NH), 3075 (Ar-CH), 2920 (CH2-CH), 1681 (C=O), 1657 (C=N), 1589 (C=C), 1056 (C-O-C). 1H NMR (CDCl3, 500 MHz) δ ppm: 2.70-4.29 (m, 8H, CH2 of morpholine), 4.55 (s, 2H, CH2 linkage), 5.23 (s, 2H, NH2), 6.86-8.12 (m, 8H, Ar-H), 8.61 (s, 1H, NH of hydrazide). 13C-NMR (δ: ppm): 51.5 (C-2’ & C-6’), 67.9 (C-3’ & C-5’), 71.3 (CH2 linkage), 116.1 (C-8), 120.6 (C-7), 123.2 (C-2” & C-6”), 124.4 (C-5), 127.5 (C-3” & C-5”), 128.0 (C-4), 130.8 (C-6), 134.8 (C-4”), 145.6 (C-9), 155.3 (C-1”), 161.8 (C-3), 162.1 (C-2), 163.5 (CONHNH2). EI-MS m/z: 379 (M+). Anal. Cald for C20H21N5O3: C, 63.31; H, 5.58; N, 18.46. Found: C, 63.50; H, 5.57; N, 18.48.
4-(2-Oxo-1-(piperidin-1-ylmethyl)indolin-3-ylideneamino)benzo hydrazide (4b)
Yield: 75 % (2.83 g), m.p.: 181-183 °C, Rf Value: 0.60 [Hexane: Chloroform: n-Butanol (40: 20: 40)]. IR (KBr, cm‒1): 3362 & 3319 (NH), 3056 (Ar-CH), 2928 (CH2-CH), 1674 (C=O), 1641 (C=N), 1604 (C=C). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.73-2.96 (m, 10H, CH2 of piperidine), 4.62 (s, 2H, CH2 linkage), 5.08 (s, 2H, NH2), 7.25-8.31 (m, 8H, Ar-H), 8.94 (s, 1H, NH of hydrazide). 13C-NMR (δ: ppm): 25.4 (C-3’ & C-5’), 25.9 (C-4’), 52.1 (C-2’ & C-6’), 72.5 (CH2 linkage), 117.6 (C-8), 121.3 (C-7), 123.8 (C-2” & C-6”), 125.2 (C-5), 128.2 (C-3” & C-5”), 130.7 (C-4), 132.4 (C-6), 133.5 (C-4”), 146.8 (C-9), 154.0 (C-1”), 164.2 (C-3), 164.8 (C-2), 165.3 (CONHNH2). EI-MS m/z: 377 (M+). Anal. Cald for C21H23N5O2: C, 66.83; H, 6.14; N, 18.55. Found: C, 66.64; H, 6.16; N, 18.52.
Synthesis of 3-(4-(5-(substituted phenyl)-3-phenyl-4,5-dihydro-1H-pyrazole-1-carbonyl) phenylimino)-1-(substitutedmethyl)indolin-2-one (5a-5l)
Equimolar quantity of various aromatic/heterocyclic aldehydes (0.01 mol) and acetophenone (1.20 g; 0.01 mol) was dissolved in minimum quantity of ethanol. To this mixture, a catalytic quantity of sodium hydroxide pellet was added. Then the reaction mixture was stirred for 2-8 h and poured on ice cold water. The product separated (chalcone) was filtered, dried and utilized for further step. Various aromatic chalcone (0.01 mol) was added to the 4-(1-(substitutedmethyl)-2-oxoindolin-3-ylideneamino)benzohydrazide 4a/4b (3.79/3.77 g; 0.01 mol) in round bottom flask containing N,N-dimethyl formamide (50 ml). The above mixture was refluxed in oil bath for a period of 12 h. Then the reaction mixture was cooled, poured in to a beaker containing ice cold water, and kept aside for 24 h. The obtained product 5a-5l was separated by filtration, dried over the filter paper, and recrystallised using ethanol.
3-(4-(5-(4-Chlorophenyl)-3-phenyl-4,5-dihydro-1H-pyrazole-1-carbonyl) phenylimino)-1-(morpholinomethyl)indolin-2-one (5a)
Yield: 71 % (4.28 g), m.p.: 265-267 °C, Rf Value: 0.51 [Hexane: Chloroform: n-Butanol (20: 30: 50)]. IR (KBr, cm‒1): 3024 (Ar-CH), 2971 (CH2-CH), 1740 (C=O), 1677 (C=N), 1591 (C=C), 1029 (C-O-C), 756(C-Cl). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.84 (d, 2H, CH2 of pyrazole), 2.31-2.87 (m, 8H, CH2 of morpholine), 4.26 (s, 2H, CH2 linkage), 5.14 (t, 1H, CH of pyrazole), 6.93-8.25 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 38.4 (CH2 of pyrazole), 52.6 (C-2’ & C-6’), 59.5 (C-5 of pyrazole), 65.8 (C-3’ & C-5’), 70.6 (CH2 linkage), 115.2 (C-8), 122.3 (C-7), 123.0 (C-2” & C-6”), 125.9 (C-5), 130.7 (C-2”’ & C-6”’), 130.9 (C-3”’ & C-5”’), 131.0 (C-3” & C-5”), 131.3 (C-3”” & C-5””), 131.9 (C-2”” & C-6””), 132.1 (C-4), 132.5 (C-4””), 133.0 (C-6), 133.2 (C-4”’), 134.5 (C-4”), 135.7 (C-1””), 140.2 (C-1”’), 146.4 (C-9), 150.4 (C-3 of pyrazole), 154.6 (C-1”), 166.2 (C-3), 166.1 (C-2), 170.8 (CO). EI-MS m/z: 603 (M+), 605 (M+2). Anal. Cald for C35H30ClN5O3: C, 69.59; H, 5.01; N, 11.59. Found: C, 70.25; H, 4.99; N, 11.56.
1-(Morpholinomethyl)-3-(4-(3-phenyl-5-(4-(trifluoromethyl)phenyl)-4,5-dihydro-1H-pyrazole-1-carbonyl)phenylimino)indolin-2-one (5b)
Yield: 79 % (5.03 g), m.p.: 209-212 °C, Rf Value: 0.84 [Hexane: Chloroform: n-Butanol (20: 30: 50)]. IR (KBr, cm‒1): 3072 (Ar-CH), 2939 (CH2-CH), 1681 (C=O), 1654 (C=N), 1596 (C=C), 1135 (C-O-C), 1127(C-F). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.87 (d, 2H, CH2 of pyrazole), 2.52-3.04 (m, 8H, CH2 of morpholine), 4.11 (s, 2H, CH2 linkage), 5.08 (t, 1H, CH of pyrazole), 7.13-8.22 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 40.1 (CH2 of pyrazole), 50.6 (C-2’ & C-6’), 60.2 (C-5 of pyrazole), 64.5 (C-3’ & C-5’), 71.0 (CH2 linkage), 116.2 (C-8), 120.8 (C-7), 121.0 (C-2” & C-6”), 125.3 (CF3), 125.8 (C-5), 126.0 (C-3”’ & C-5”’), 127.4 (C-2”’ & C-6”’), 129.3 (C-3” & C-5”), 129.6 (C-3”” & C-5””), 129.8 (C-2”” & C-6””), 130.1 (C-4), 130.4 (C-4””), 131.2 (C-6), 129.5 (C-4”’), 133.9 (C-4”), 135.3 (C-1””), 145.1 (C-1”’), 148.7 (C-9), 153.1 (C-3 of pyrazole), 156.8 (C-1”), 165.0 (C-3), 165.4 (C-2), 169.5 (CO). EI-MS m/z: 637 (M+). Anal. Cald for C36H30F3N5O3: C, 67.81; H, 4.74; N, 10.98. Found: C, 67.63; H, 4.73; N, 11.02.
1-(Morpholinomethyl)-3-(4-(5-(4-nitrophenyl)-3-phenyl-4,5-dihydro-1H-pyrazole-1-carbonyl)phenylimino)indolin-2-one (5c)
Yield: 76 % (4.67 g), m.p.: 234-236 °C, Rf Value: 0.47 [Hexane: Chloroform: n-Butanol (20: 30: 50)]. IR (KBr, cm‒1): 3056 (Ar-CH), 2943 (CH2-CH), 1687 (C=O), 1642 (C=N), 1588 (C=C), 1550 & 1321 (NO2), 1119 (C-O-C). 1H NMR (CDCl3, 500 MHz) δ ppm: 2.10 (d, 2H, CH2 of pyrazole), 2.44-2.91 (m, 8H, CH2 of morpholine), 4.23 (s, 2H, CH2 linkage), 5.25 (t, 1H, CH of pyrazole), 7.09-8.07 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 37.3 (CH2 of pyrazole), 50.2 (C-2’ & C-6’), 57.7 (C-5 of pyrazole), 67.6 (C-3’ & C-5’), 69.3 (CH2 linkage), 118.5 (C-8), 120.1 (C-3”’ & C-5”’), 121.9 (C-7), 123.6 (C-2” & C-6”), 125.4 (C-5), 127.0 (C-2”’ & C-6”’), 129.4 (C-3” & C-5”), 129.7 (C-3”” & C-5””), 130.4 (C-2”” & C-6””), 131.0 (C-4), 132.9 (C-4””), 133.5 (C-6), 133.6 (C-4”), 135.2 (C-1””), 146.8 (C-4”’), 149.3 (C-9), 151.8 (C-1”’), 152.3 (C-3 of pyrazole), 159.4 (C-1”), 164.5 (C-3), 165.6 (C-2), 168.0 (CO). EI-MS m/z: 614 (M+). Anal. Cald for C35H30N6O5: C, 68.39; H, 4.92; N, 13.67. Found: C, 68.53; H, 4.93; N, 13.64.
1-(Morpholinomethyl)-3-(4-(3-phenyl-5-p-tolyl-4,5-dihydro-1H-pyrazole-1-carbonyl) phenylimino)indolin-2-one (5d)
Yield: 73 % (4.26 g), m.p.: 250-252 °C, Rf Value: 0.55 [Hexane: Chloroform: n-Butanol (20: 30: 50)]. IR (KBr, cm‒1): 3068 (Ar-CH), 2951 (CH2-CH), 1665 (C=O), 1647 (C=N), 1594 (C=C), 1120 (C-O-C). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.83 (d, 2H, CH2 of pyrazole), 2.12 (s, 3H, CH3), 2.58-3.16 (m, 8H, CH2 of morpholine), 4.30 (s, 2H, CH2 linkage), 5.09 (t, 1H, CH of pyrazole), 6.84-7.91 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 26.7 (CH3), 39.4 (CH2 of pyrazole), 49.7 (C-2’ & C-6’), 56.1 (C-5 of pyrazole), 65.0 (C-3’ & C-5’), 73.9 (CH2 linkage), 114.5 (C-8), 118.2 (C-7), 120.8 (C-2” & C-6”), 123.6 (C-5), 126.3 (C-2”’ & C-6”’), 128.1 (C-3”’ & C-5”’), 127.5 (C-3” & C-5”), 127.7 (C-3”” & C-5””), 129.0 (C-2”” & C-6””), 129.2 (C-4), 130.3 (C-4””), 132.6 (C-6), 135.9 (C-4”’), 133.4 (C-4”), 133.8 (C-1””), 142.7 (C-1”’), 147.5 (C-9), 150.6 (C-3 of pyrazole), 158.1 (C-1”), 162.9 (C-3), 163.8 (C-2), 167.3 (CO). EI-MS m/z: 583 (M+). Anal. Cald for C36H33N5O3: C, 74.08; H, 5.70; N, 12.00. Found: C, 74.26; H, 5.68; N, 11.97.
3-(4-(5-(4-Aminophenyl)-3-phenyl-4,5-dihydro-1H-pyrazole-1-carbonyl)phenylimino)-1-(morpholinomethyl)indolin-2-one (5e)
Yield: 78 % (4.56 g), m.p.: 197-199 °C, Rf Value: 0.42 [Hexane: Chloroform: n-Butanol (20: 30: 50)]. IR (KBr, cm‒1): 3301 (NH), 3023 (Ar-CH), 2946 (CH2-CH), 1740 (C=O), 1668 (C=N), 1631 (C=C), 1110 (C-O-C). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.91 (d, 2H, CH2 of pyrazole), 2.70-3.23 (m, 8H, CH2 of morpholine), 4.02 (s, 2H, NH2), 4.63 (s, 2H, CH2 linkage), 5.49 (t, 1H, CH of pyrazole), 7.21-8.68 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 39.7 (CH2 of pyrazole), 51.3 (C-2’ & C-6’), 60.8 (C-5 of pyrazole), 66.1 (C-3’ & C-5’), 72.4 (CH2 linkage), 114.3 (C-3”’ & C-5”’), 117.8 (C-8), 119.5 (C-7), 120.4 (C-2” & C-6”), 123.0 (C-5), 126.7 (C-2”’ & C-6”’), 129.5 (C-3” & C-5”), 130.2 (C-3”” & C-5””), 130.6 (C-2”” & C-6””), 130.9 (C-4), 132.4 (C-4””), 132.8 (C-6), 133.1 (C-4”), 134.0 (C-1”’), 135.6 (C-1””), 144.3 (C-4”’), 148.9 (C-9), 151.9 (C-3 of pyrazole), 156.1 (C-1”), 161.3 (C-3), 161.4 (C-2), 166.2 (CO). EI-MS m/z: 584 (M+). Anal. Cald for C35H32N6O3: C, 71.90; H, 5.52; N, 14.37. Found: C, 71.68; H, 5.54; N, 14.41.
3-(4-(5-(4-Hydroxyphenyl)-3-phenyl-4,5-dihydro-1H-pyrazole-1-carbonyl)phenylimino)-1-(morpholinomethyl)indolin-2-one (5f)
Yield: 80 % (4.68 g), m.p.: 228-230 °C, Rf Value: 0.69 [Hexane: Chloroform: n-Butanol (20: 30: 50)]. IR (KBr, cm‒1): 3557 (OH), 3040 (Ar-CH), 2934 (CH2-CH), 1679 (C=O), 1665 (C=N), 1599 (C=C), 1136 (C=O). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.86 (d, 2H, CH2 of pyrazole), 2.49-3.03 (m, 8H, CH2 of morpholine), 4.25 (s, 2H, CH2 linkage), 5.24 (t, 1H, CH of pyrazole), 5.61 (s, 1H, OH), 7.01-8.37 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 37.2 (CH2 of pyrazole), 54.9 (C-2’ & C-6’), 57.0 (C-5 of pyrazole), 68.3 (C-3’ & C-5’), 70.5 (CH2 linkage), 112.4 (C-3”’ & C-5”’), 118.7 (C-8), 122.3 (C-7), 123.2 (C-2” & C-6”), 126.1 (C-5), 130.6 (C-2”’ & C-6”’), 131.2 (C-3” & C-5”), 132.5 (C-3”” & C-5””), 132.8 (C-2”” & C-6””), 133.0 (C-4), 133.7 (C-4””), 134.3 (C-6), 134.6 (C-4”), 135.4 (C-1””), 137.1 (C-1”’), 149.0 (C-9), 152.5 (C-3 of pyrazole), 154.8 (C-4”’), 155.9 (C-1”), 163.1 (C-3), 164.0 (C-2), 168.6 (CO). EI-MS m/z: 585 (M+). Anal. Cald for C35H31N5O4: C, 71.78; H, 5.34; N, 11.96. Found: C, 71.56; H, 5.35; N, 11.99.
3-(4-(5-(4-Chlorophenyl)-3-phenyl-4,5-dihydro-1H-pyrazole-1-carbonyl)phenylimino)-1-(piperidin-1-ylmethyl)indolin-2-one (5g)
Yield: 75 % (4.51 g), m.p.: 293-295 °C, Rf Value: 0.53 [Hexane: Chloroform: n-Butanol (40: 20: 40)]. IR (KBr, cm‒1): 3076 (Ar-CH), 2957 (CH2-CH), 1683 (C=O), 1658 (C=N), 1570 (C=C), 735 (C-Cl). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.92 (d, 2H, CH2 of pyrazole), 2.57-3.39 (m, 10H, CH2 of piperidine), 4.26 (s, 2H, CH2 linkage), 5.31 (t, 1H, CH of pyrazole), 7.25-8.10 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 24.9 (C-3’ & C-5’), 25.3 (C-4’), 38.1 (CH2 of pyrazole), 53.9 (C-2’ & C-6’), 58.4 (C-5 of pyrazole), 71.7 (CH2 linkage), 115.3 (C-8), 120.5 (C-7), 121.9 (C-2” & C-6”), 124.5 (C-5), 127.2 (C-2”’ & C-6”’), 129.0 (C-3”’ & C-5”’), 129.8 (C-3” & C-5”), 130.0 (C-3”” & C-5””), 130.7 (C-2”” & C-6””), 131.0 (C-4), 131.1 (C-4””), 131.4 (C-6), 133.5 (C-4”), 133.9 (C-4”’), 136.2 (C-1””), 143.4 (C-1”’), 147.6 (C-9), 150.7 (C-3 of pyrazole), 157.0 (C-1”), 162.3 (C-3), 162.7 (C-2), 168.9 (CO). EI-MS m/z: 601 (M+), 603 (M+2). Anal. Cald for C36H32ClN5O2: C, 71.81; H, 5.36; N, 11.63. Found: C, 72.04; H, 5.34; N, 11.59.
3-(4-(3-Phenyl-5-(4-(trifluoromethyl)phenyl)-4,5-dihydro-1H-pyrazole-1-carbonyl) phenylimino)-1-(piperidin-1-ylmethyl)indolin-2-one (5h)
Yield: 72 % (4.57 g), m.p.: 152-154 °C, Rf Value: 0.45 [Hexane: Chloroform: n-Butanol (40: 20: 40)]. IR (KBr, cm‒1): 3059 (Ar-CH), 2946 (CH2-CH), 1678 (C=O), 1641 (C=N), 1585 (C=C), 1142 (C-F). 1H NMR (CDCl3, 500 MHz) δ ppm: 2.09 (d, 2H, CH2 of pyrazole), 2.51-3.48 (m, 10H, CH2 of piperidine), 4.12 (s, 2H, CH2 linkage), 5.17 (t, 1H, CH of pyrazole), 7.05-8.25 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 23.2 (C-3’ & C-5’), 23.6 (C-4’), 38.0 (CH2 of pyrazole), 52.4 (C-2’ & C-6’), 62.9 (C-5 of pyrazole), 69.8 (CH2 linkage), 119.5 (C-8), 121.8 (C-7), 122.1 (C-2” & C-6”), 123.0 (CF3), 123.5 (C-5), 127.2 (C-3”’ & C-5”’), 129.7 (C-2”’ & C-6”’), 130.2 (C-3” & C-5”), 130.4 (C-3”” & C-5””), 130.5 (C-4”’), 131.3 (C-2”” & C-6””), 132.1 (C-4), 133.3 (C-4””), 133.6 (C-6), 134.0 (C-4”), 135.5 (C-1””), 148.7 (C-1”’), 151.5 (C-9), 154.8 (C-3 of pyrazole), 157.5 (C-1”), 161.2 (C-3), 161.8 (C-2), 168.4 (CO). EI-MS m/z: 635 (M+). Anal. Cald for C37H32F3N5O2: C, 69.91; H, 5.07; N, 11.02. Found: C, 69.73; H, 5.09; N, 11.05.
3-(4-(5-(4-Nitrophenyl)-3-phenyl-4,5-dihydro-1H-pyrazole-1-carbonyl)phenylimino)-1-(piperidin-1-ylmethyl)indolin-2-one (5i)
Yield: 77 % (4.71 g), m.p.: 190-192 °C, Rf Value: 0.71 [Hexane: Chloroform: n-Butanol (40: 20: 40)]. IR (KBr, cm‒1): 3071 (Ar-CH), 2948 (CH2-CH), 1674 (C=O), 1649 (C=N), 1582 (C=C), 1538 & 1343 (NO2). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.98 (d, 2H, CH2 of pyrazole), 2.65-3.29 (m, 10H, CH2 of piperidine), 4.36 (s, 2H, CH2 linkage), 5.22 (t, 1H, CH of pyrazole), 6.90-8.04 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 25.8 (C-3’ & C-5’), 25.9 (C-4’), 40.7 (CH2 of pyrazole), 50.1 (C-2’ & C-6’), 61.4 (C-5 of pyrazole), 68.4 (CH2 linkage), 117.0 (C-8), 120.5 (C-3”’ & C-5”’), 122.6 (C-7), 124.3 (C-2” & C-6”), 126.2 (C-5), 127.5 (C-2”’ & C-6”’), 128.3 (C-3” & C-5”), 128.7 (C-3”” & C-5””), 129.8 (C-2”” & C-6””), 130.2 (C-4), 130.9 (C-4””), 131.7 (C-6), 134.1 (C-4”), 136.0 (C-1””), 145.2 (C-4”’), 150.9 (C-9), 151.5 (C-1”’), 153.4 (C-3 of pyrazole), 154.9 (C-1”), 165.0 (C-3), 165.3 (C-2), 170.7 (CO). EI-MS m/z: 612 (M+). Anal. Cald for C36H32N6O4: C, 70.57; H, 5.26; N, 13.72. Found: C, 70.78; H, 5.27; N, 13.69.
3-(4-(3-Phenyl-5-p-tolyl-4,5-dihydro-1H-pyrazole-1-carbonyl) phenylimino)-1-(piperidin-1-ylmethyl)indolin-2-one (5j)
Yield: 74 % (4.30 g), m.p.: 281-284 °C, Rf Value: 0.49 [Hexane: Chloroform: n-Butanol (40: 20: 40)]. IR (KBr, cm‒1): 3015 (Ar-CH), 2952 (CH2-CH), 1666 (C=O), 1643 (C=N), 1597 (C=C). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.80 (d, 2H, CH2 of pyrazole), 2.28 (s, 3H, CH3), 2.46-3.02 (m, 10H, CH2 of piperidine), 4.14 (s, 2H, CH2 linkage), 5.27 (t, 1H, CH of pyrazole), 7.11-8.36 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 22.4 (CH3), 23.7 (C-3’ & C-5’), 24.0 (C-4’), 41.9 (CH2 of pyrazole), 51.5 (C-2’ & C-6’), 58.6 (C-5 of pyrazole), 73.2 (CH2 linkage), 117.5 (C-8), 120.4 (C-7), 123.8 (C-2” & C-6”), 125.3 (C-5), 126.1 (C-2”’ & C-6”’), 127.5 (C-3”’ & C-5”’), 127.8 (C-3” & C-5”), 128.0 (C-3”” & C-5””), 128.6 (C-2”” & C-6””), 129.2 (C-4), 129.7 (C-4””), 131.5 (C-6), 132.4 (C-4”), 135.1 (C-1””), 136.8 (C-4”’), 143.4 (C-1”’), 148.6 (C-9), 149.2 (C-3 of pyrazole), 156.7 (C-1”), 164.5 (C-3), 164.7 (C-2), 167.9 (CO). EI-MS m/z: 581 (M+). Anal. Cald for C37H35N5O2: C, 76.40; H, 6.06; N, 12.04. Found: C, 76.25; H, 6.08; N, 12.03.
3-(4-(5-(4-Aminophenyl)-3-phenyl-4,5-dihydro-1H-pyrazole-1-carbonyl) phenylimino)-1-(piperidin-1-ylmethyl)indolin-2-one (5k)
Yield: 71 % (4.13 g), m.p.: 257-259 °C, Rf Value: 0.80 [Hexane: Chloroform: n-Butanol (40: 20: 40)]. IR (KBr, cm‒1): 3324 (NH), 3067 (Ar-CH), 2935 (CH2-CH), 1682 (C=O), 1676 (C=N), 1573 (C=C). 1H NMR (CDCl3, 500 MHz) δ ppm: 1.95 (d, 2H, CH2 of pyrazole), 2.53-3.35 (m, 10H, CH2 of piperidine), 3.97 (s, 2H, NH2), 4.28 (s, 2H, CH2 linkage), 5.36 (t, 1H, CH of pyrazole), 6.82-8.13 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 26.5 (C-3’ & C-5’), 27.1 (C-4’), 39.7 (CH2 of pyrazole), 51.3 (C-2’ & C-6’), 59.0 (C-5 of pyrazole), 70.7 (CH2 linkage), 113.2 (C-3”’ & C-5”’), 119.2 (C-8), 120.9 (C-7), 121.4 (C-2” & C-6”), 124.9 (C-5), 126.7 (C-2”’ & C-6”’), 127.6 (C-3” & C-5”), 127.8 (C-3”” & C-5””), 128.1 (C-2”” & C-6””), 128.6 (C-4), 128.9 (C-4””), 129.3 (C-6), 131.7 (C-4”), 132.5 (C-1”’), 133.4 (C-1””), 147.1 (C-4”’), 151.9 (C-9), 152.5 (C-3 of pyrazole), 157.3 (C-1”), 161.0 (C-3), 162.7 (C-2), 168.1 (CO). EI-MS m/z: 582 (M+). Anal. Cald for C36H34N6O2: C, 74.20; H, 5.88; N, 14.42. Found: C, 74.03; H, 5.86; N, 14.47.
3-(4-(5-(4-Hydroxyphenyl)-3-phenyl-4,5-dihydro-1H-pyrazole-1-carbonyl)phenylimino)-1-(piperidin-1-ylmethyl)indolin-2-one (5l)
Yield: 76 % (4.43 g), m.p.: 222-224 °C, Rf Value: 0.38 [Hexane: Chloroform: n-Butanol (40: 20: 40)]. IR (KBr, cm‒1): 3528 (OH), 3023 (Ar-CH), 2947 (CH2-CH), 1740 (C=O), 1667 (C=N), 1629 (C=C). 1H NMR (CDCl3, 500 MHz) δ ppm: 2.01 (d, 2H, CH2 of pyrazole), 2.61-3.27 (m, 10H, CH2 of piperidine), 4.11 (s, 2H, CH2 linkage), 4.70 (t, 1H, CH of pyrazole), 5.31 (s, 1H, OH), 7.01-8.54 (m, 17H, Ar-H). 13C-NMR (δ: ppm): 24.1 (C-3’ & C-5’), 24.5 (C-4’), 42.6 (CH2 of pyrazole), 50.7 (C-2’ & C-6’), 62.3 (C-5 of pyrazole), 71.4 (CH2 linkage), 114.7 (C-3”’ & C-5”’), 116.3 (C-8), 121.9 (C-7), 122.9 (C-2” & C-6”), 126.2 (C-5), 129.8 (C-2”’ & C-6”’), 130.0 (C-3” & C-5”), 130.1 (C-3”” & C-5””), 130.9 (C-2”” & C-6””), 131.5 (C-4), 131.8 (C-4””), 132.4 (C-6), 133.6 (C-4”), 134.9 (C-1””), 139.5 (C-1”’), 150.2 (C-9), 151.4 (C-3 of pyrazole), 154.3 (C-4”’), 155.2 (C-1”), 162.7 (C-3), 163.1 (C-2), 166.8 (CO). EI-MS m/z: 583 (M+). Anal. Cald for C36H33N5O3: C, 74.08; H, 5.70; N, 12.00. Found: C, 74.36; H, 5.71; N, 11.96.
CHEMISTRY
A range of novel pyrazole substituted isatin derivatives were synthesized by multistep synthesis from indole-2,3-dione in the present study. Initially, isatin 1 was treated with morpholine/piperidine and formaldehyde to produce 1-(substitutedmethyl)indoline-2,3-dione 2a-2b by Mannich reaction. In the succeeding stair, 4-(1-(substitutedmethyl)-2-oxoindolin-3-ylideneamino)benzoic acid 3a-3b was synthesized through the condensation of compound 2a-2b with p-aminobenzoic acid in ethanol by Schiff base reaction. On treating with thionyl chloride followed by hydrazine hydrate, compounds 3a-3b converted to its respective hydrazide derivatives [4-(1-(substitutedmethyl)-2-oxoindolin-3-ylideneamino)benzohydrazide] 4a-4b. In the final step, the title compounds 5a-5l were synthesized by a ring closure reaction, in which a different chalcones (α,β-unsaturated carbonyl compound) and hydrazide derivative of isatin derivatives 4a-4b were reacted. TLC was performed throughout the reactions to optimize the reactions for purity and completion.
IR, 1H-NMR, Mass spectra, and elemental analyses of the synthesized compounds are in accordance with the assigned structures. The IR spectra of all synthesized compounds showed some characteristic peaks indicating the presence of particular groups. Formation of the methylene linkage in compound 2a/2b was confirmed by its absorption peak at 2937/2922 cm-1 in IR corresponds to CH2-CH stretching respectively. This was further confirmed by appearance of singlet for two protons of CH2 linkage at δ 4.96 and 4.53 ppm for compound 2a and 2b respectively. The absorption bands at 3405-3422 and 1668-1681cm-1, which can be assignable to OH of COOH and C=N (azomethine linkage) vibrations respectively confirms the formation of compounds 3a-3b. Appearance of singlet at δ 9.94-10.09 ppm for single protons in its 1H-NMR spectra which might be assigned to OH of COOH group additionally support the structure of compounds 3a-3b. The conversion of hydrazide 4a-4b from carboxylic acid 3a-3b can be recognized by strong absorption peak at 3283-3362 cm-1 in IR due to N-H stretching and appearance of singlet in its 1H-NMR spectra at δ 5.08-5.23 ppm for two protons which might be assigned to NH2 of hydrazide group. Presence of NH in hydrazide was confirmed by appearance of singlet for single proton in 1H-NMR spectra at δ 8.61-8.94 ppm. The structure of title compounds 5a-5l was confirmed by the absence of absorption band around 3300 cm-1 due to absence of NH stretching vibration. In addition appearance of doublet for two protons of C-4 pyrazole at δ 1.80-2.10 ppm and triplet for one proton of C-5 pyrazole at δ 4.70-5.49 ppm also confirms the structure of title compounds 5a-5l. The IR spectrum of title compounds 5a-5l shows absorption bands at 3015-3076 cm-1, 2934-2971 cm-1, 1665-1740 cm-1, 1641-1677 cm-1 and 1570-1631 cm-1, which can be assignable to Ar-CH, CH2-CH, C=O, C=N and C=C vibrations respectively. The following conclusions can be derived by comparing the NMR spectra of synthesized compounds 5a-5l: a) A doublet at δ 1.80-2.10 ppm for CH2 of pyrazole, b) A multiplet at δ 2.31-3.48 ppm for CH2 of morpholine/piperidine, c) A singlet at δ 4.11-4.63 ppm for CH2 linkage, d) A triplet at δ 4.70-5.49 ppm for CH of pyrazole, e) A multiplet at δ 6.82-8.68 ppm for Ar-H. Further mass spectrum confirmed their molecular weight. Elemental analysis of all synthesized compounds was found to be within + 0.4 % with respect to calculated value which confirmed the purity of synthesized compounds.