Chemistry
In the research work, we have synthesized new series of N-substitutedbenzylidene-5-(4-formylphenyl)-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide analogues using direct condensation of aromatic aldehydes and substituted acetophenone. Synthetic steps of this series have been shown in Scheme-1. The structure of synthesized derivatives was examined by 1H-NMR, FT-IR, MS and Elemental analysis. In Table 1, physiochemical properties of pyrazole derivatives have been presented. The physiochemical properties included molecular formula, molecular weight, melting points, percentage yield and Rf value of synthesized analogues.
Antioxidant activity
The antioxidant activity of the newly synthesized analogues was examined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay using free radical scavenging method (spectrophotometrically). DPPH, on reaction with hydrogen donors, a free radical with maximal absorption at 517 nm is reduced to hydrazine (corresponding compound) indicating deep violet color of DPPH change to yellow, showing a considerable decrease in absorption. DPPH solution (3 μg/ml) was prepared in methanol and DPPH (in 1:1) solution was used for blank reference. 25 μg/ml, 50 μg/ml, 75 μg/ml, 100 μg/ml of each synthesized analogues and ascorbic acid (standard) was prepared in methanol. Then, equal amount of each concentration and DPPH was taken in a test tube. Afterwards, absorbance was taken at 517 nm by UV spectrophotometer of the solution mixture by stirring it for 5 min and stored at dark place for half an hour at room temperature [18]. The % inhibition was evaluated as follow
Where,
AControl= absorbance of the control reaction.
ASample= absorbance of the test compound.
IC50 value was calculated from the graph shown in Figures 3-5 which indicates % inhibition and synthesized compounds. The results of antioxidant activity explained that compounds Y9 and Y17 having maximum in vitro antioxidant potency with IC50 values 17.43 mol/L and 18.98 mol/L, in comparison with ascorbic acid (standard drug). The presented results are showing in Table 2.
Antidiabetic activity
All the synthesized derivatives were investigate for the α-amylase inhibitory activity by using diastase based on colorimetric method [19]. For preparing enzyme solution, 1 mg diastase (amylase enzyme) was taken in 100 mL of 20 mM phosphate buffer. 25 μg/ml, 50 μg/ml, 75 μg/ml, 100 μg/ml concentration of synthesized compounds was prepared in DMSO. 0.25g potato starch was taken and adding in 50 mL of 20 mM phosphate buffer for potato starch solution by heating 15 min. The colour reagent was prepared by mixing 20 mL of 96 mM 3,5 dinitrosalicylic acid with 5.31 M sodium potassium tartrate in 8 mL of 2 M sodium hydroxide and 12 mL distilled water. After preparation of valuable solutions that needed for activity, 1 mL of synthesized derivatives solution and 1 mL of enzyme solution incubated in 25 °C temp. for 10 min. After that taken 1 mL of this mixture and added 1 mL of potato starch solution in test tube, incubated in 25 °C temp. for 10 min. Then test tubes was closed after adding 1 mL colour reagent and placed at water bath at 85 °C for 15 min. When reaction mixture was cooled diluted with 9 mL of distilled water and taken absorbance at 540 nm in UV spectrophotometer. % inhibition was calculated as follow.
IC50 value was calculated from the graph shown in Figures 6-8 which indicates % inhibition and synthesized compound. The evaluation of antidiabetic activity explain that compounds Y1, Y5 and Y6 exhibited excellent antidiabetic potency with IC50 values 17.08 mol/L, 08.36 mol/L and 13.50 mol/L respectively, in comparison with acarbose (standard drug). The presented results are showing in Table 3.
Anti-inflammatory activity
All the synthesized derivatives were investigated for the protein albumin denaturation [20,21]. Different concentration of test compounds was prepared (10 μg/ml, 30 μg/ml, 50 μg/ml, 70 μg/ml and 100 μg/ml) in methanol. After preparation of test samples 1 % aqueous solution of bovine albumin serium in 0.05 M tris buffer saline and pH (7.6 at 25 °C) of reaction mixture was adjusted by adding small amount of 1 N HCl. The blank solution was taken is methanol. Then equal amount of test and protein was taken in the test tube and incubated for 20 min. at 37 °C. After incubation the reaction mixture was heated for 20 min. at 57 °C temperature and cooling the reaction mixture, the turbidity was measured at 660 nm in UV spectrophotometer. Percent inhibition of protein denaturation was calculated as follows:
IC50 value was calculated from the graph shown in Figures 9-11 which indicates % inhibition and synthesized compound. The results of anti-inflammatory activity showed that compounds Y2, Y3 and Y7 exhibited excellent anti-inflammatory activity with IC50 values 23.23 mol/L, 22.09 mol/L and 19.05 mol/L respectively, in comparison with aspirin (standard drug). The presented results are showing in Table 4.
SAR (Structure Activity Relationship) studies:
From the antioxidant, antidiabetic and anti-inflammatory testing evaluation of newly synthesized N-substitutedbenzylidene-5-(4-formylphenyl)-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide derivatives, the ensuing structure activity relationship showed in fig 12.
- Presence of electron withdrawing and releasing group (-OH and -Cl, Compounds Y9 and Y17) on benzylidene portion enhanced the antioxidant potency of the synthesized compounds.
- Presence of electron withdrawing and releasing group (-CHO and -OCH3, Compounds Y1, Y5 and Y6) on benzylidene portion enhanced the antidiabetic activity and (Compounds Y2, Y3and Y7) anti-inflammatory activity of the synthesized compounds.
Experimental section
In Scheme-1, synthetic procedure of pyrazole derivatives has been explained. The scheme was drawn via ChemDraw Ultra 8.03. The completion of reaction was examined by TLC (Thin Layer Chromatography) by using silica gel glass plate. Open capillary tube method was used to examine melting point of synthesized analogues. Attenuated total reflection (ATR-IR) spectrophotometer of Bruker FTIR 12060280 (Software: OPUS 7.2.139.1294) was used for evaluating chemical structure of synthesized analogues by recording IR spectra. Bruker Avance III 600 NMR spectrometer was used for 1H/13CNMR (Nuclear Magnetic Resonance). The spectra was recorded in ppm (δ) with suitable deuterated solvent like DMSO and internal solvent like tetramethyl silane and data have shown s, singlet; d, douplet; t, triplet; m, multiplet (multiplicity), number of proton and carbon values. Mass spectra were recorded by Waters Micromass Q-ToF Micro spectrophotometer. Perkin-Elmer 2400 was used for elemental analysis.
General procedure for the synthesis of N-substitutedbenzylidene-5-(4-formylphenyl)-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide derivatives (Y1-Y26)
Step I: Synthesis of 4-(3-(4-hydroxyphenyl)-3-oxoprop-1-enyl)benzaldehyde (intermediate 1)
An equal quantity of 4-hydroxy acetophenone and terepthaldehyde (0.01 mol) was taken in a 20 mL ethanol and stirred for few minutes until it get mixed. A 10 mL fraction of 40% aqueous solution of KOH was prepared and slowly mixed drop wise in a reaction mixture. The reaction mixture stirred for a day at room temperature. When reaction was completed, the reaction mixture was acidified with diluted HCl and precipitate was formed, collected by filtration. The completion of reaction was monitored by using TLC.
Step II: Synthesis of 3,5-bis(4-hydroxyphenyl)-4,5-dihydropyrazole-1-caebothioamide (intermediate 2)
A mixture of intermediate 1 (0.01 mol), thiosemicarbazide (0.01 mol) and KOH (0.0025 mol) in 50 mL ethanol were taken in RBF and refluxed for 72 hours. When reaction was completed, the reaction mixture was acidified with HCl and to yield solid precipitate, filtered, washed out by water, dried and recrystallized with ethanol. By using TLC, reaction completion was examined [22].
Step III: Synthesis of N-substitutedbenzylidene-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide derivatives
A mixture of intermediate 2 (0.01 mol), substituted benzaldehyde (0.01mol) in 50 mL ethanol were taken in RBF and refluxed for some time then 1 mL glacial acetic acid was added drop wise in a reaction mixture and refluxed for 48 hours. When reaction was completed, the resulting mixture was cooled, poured in water to yield solid precipitate, filtered, dried and recrystallized with ethanol. By using TLC, reaction completion was examined.
Spectral data of synthesized pyrimidine derivatives
Compound Y1: 5-(4-Formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3135 (C-H str., benzene), 1438 (C=C str., benzene), 1359 (C=N str., pyrazole moiety), 1088 (N-N str., pyrazole moiety), 1274 (C-N str., carbonyl), 3225 (OH str., aromatic), 1749 (CHO str., aromatic), 1217 (C=S str., aliphatic), 3012 (NH2 str., aliphatic);
1H NMR: 6.81-7.76 (s, 8H, Ar-H), 1.95-4.01 (s, 3H, pyrazole), 9.12 (s, 1H, Ar-CHO), 5.02 (s, 1H, Ar-OH), 2.12 (s, 1H, amine);13C-NMR: 40.3, 66.5, 115.2, 118.2, 121.9, 127.6, 129.8, 130.5, 134.8, 135.5, 149.5, 151.9, 168.7, 173.3, 191.5; MS ES + (ToF): m/z 326 [M++1].
Compound Y2: (E)-N-(4-Formylbenzylidene)-5-(4-formylphenyl)-3-(4-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3153 (C-H str., benzene), 1599 (C=C str., benzene), 1357 (C=N str., pyrazole moiety), 1090 (N-N str., pyrazole moiety), 1278 (C-N str., carbonyl), 3503 (OH str., aromatic), 1751 (CHO str., aromatic), 1215 (C=S str., aliphatic), 1619 (C=C str., aliphatic), 1028 (-NH- str., aliphatic); 1H NMR: 6.52-8.24 (s, 8H, Ar-H), 1.81-3.82 (s, 3H, pyrazole), 9.82 (s, 1H, Ar-CHO), 5.12 (s, 1H, Ar-OH), 8.12 (s, 1H, CH (aliphatic)); 13C-NMR: 40.5, 66.9, 115.3, 118.3, 121.7, 127.5, 129.2, 130.3, 134.9, 135.6, 149.3, 151.3, 163.4, 168.6, 187.3, 191.3; MS ES + (ToF): m/z 440 [M++1].
Compound Y3: (E)-N-(3-Formylbenzylidene)-5-(4-formylphenyl)-3-(4-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3146 (C-H str., benzene), 1592 (C=C str., benzene), 1358 (C=N str., pyrazole moiety), 1090 (N-N str., pyrazole moiety), 1274 (C-N str., carbonyl), 3202 (OH str., aromatic), 1746 (CHO str., aromatic), 1228 (C=S str., aliphatic), 1647 (C=C str., aliphatic), 1015 (-NH- str., aliphatic); 1H NMR: 6.02-8.54 (s, 8H, Ar-H), 2.08-3.42 (s, 3H, pyrazole), 9.72 (s, 1H, Ar-CHO), 5.32 (s, 1H, Ar-OH), 8.02 (s, 1H, CH (aliphatic)); 13C-NMR: 40.0, 66.3, 115.0, 118.5, 121.6, 127.9, 129.1, 130.4, 132.1, 134.8, 135.6,137.0, 149.5, 151.1, 163.5, 168.7, 187.2, 191.5; MS ES + (ToF): m/z 442 [M++1].
Compound Y4: (E)-N-(4-Hydroxy-3-methoxybenzylidene)-5-(4-formylphenyl)-3-(4-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3144 (C-H str., benzene), 1592 (C=C str., benzene), 1356 (C=N str., pyrazole moiety), 1088 (N-N str., pyrazole moiety), 1275 (C-N str., carbonyl), 3202 (OH str., aromatic), 1716 (CHO str., aromatic), 1218 (C=S str., aliphatic), 1650 (C=C str., aliphatic), 1028 (-NH- str., aliphatic), 2852 (C-OCH3 str., aromatic); 1H NMR: 6.02-8.27 (s, 8H, Ar-H), 1.01-3.18 (s, 3H, pyrazole), 9.22 (s, 1H, Ar-CHO), 5.15 (s, 1H, Ar-OH), 8.14 (s, 1H, CH (aliphatic)); 13C-NMR: 40.5, 56.5, 66.5, 114.3, 115.5, 117.2, 118.7, 121.8, 122.7, 127.6, 129.8, 130.7, 134.2, 135.8, 149.5, 151.8, 163.3, 168.9, 187.0, 191.4; MS ES + (ToF): m/z 460 [M++1].
Compound Y5: (E)-N-(3-Methoxybenzylidene)-5-(4-formylphenyl)-3-(4-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3077 (C-H str., benzene), 1600 (C=C str., benzene), 1357 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1275 (C-N str., carbonyl), 3506 (OH str., aromatic), 1752 (CHO str., aromatic), 1221 (C=S str., aliphatic), 1691 (C=C str., aliphatic), 1029 (-NH- str., aliphatic), 2845 (C-OCH3 str., aromatic); 1H NMR: 6.22-8.00 (s, 8H, Ar-H), 1.00-3.21 (s, 3H, pyrazole), 9.54 (s, 1H, Ar-CHO), 5.55 (s, 1H, Ar-OH), 8.34 (s, 1H, CH (aliphatic)); 13C-NMR: 40.9, 56.2, 66.1, 113.3, 115.5, 116.7,119.0, 121.5, 127.5, 129.8, 130.0, 134.2, 135.6, 149.3, 151.6, 160.4, 163.6, 187.9, 191.9; MS ES + (ToF): m/z 444 [M++1].
Compound Y6: (E)-N-(4-Methoxybenzylidene)-5-(4-formylphenyl)-3-(4-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3085 (C-H str., benzene), 1601 (C=C str., benzene), 1357 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1276 (C-N str., carbonyl), 3205 (OH str., aromatic), 1793 (CHO str., aromatic), 1219 (C=S str., aliphatic), 1651 (C=C str., aliphatic),1028 (-NH- str., aliphatic), 2820 (C-OCH3 str., aromatic); 1H NMR: 6.24-8.25 (s, 8H, Ar-H), 1.04-3.38 (s, 3H, pyrazole), 9.62 (s, 1H, Ar-CHO), 5.65 (s, 1H, Ar-OH), 8.54 (s, 1H, CH (aliphatic)); 13C-NMR: 44.9, 55.3, 66.7, 114.6, 115.8, 118.2, 121.8, 126.2, 127.5, 129.8, 130.9, 134.6, 135.8, 149.8, 151.8, 163.7, 187.4, 191.7; MS ES + (ToF): m/z 443 [M++1].
Compound Y7: (E)-N-(2-Methoxybenzylidene)-5-(4-formylphenyl)-3-(4-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3125 (C-H str., benzene), 1600 (C=C str., benzene), 1358 (C=N str., pyrazole moiety), 1167 (N-N str., pyrazole moiety), 1279 (C-N str., carbonyl), 3511 (OH str., aromatic), 1747 (CHO str., aromatic), 1219 (C=S str., aliphatic), 1652 (C=C str., aliphatic),1021 (-NH- str., aliphatic), 2836 (C-OCH3 str., aromatic); 1H NMR: 6.62-8.71 (s, 8H, Ar-H), 1.61-3.43 (s, 3H, pyrazole), 9.62 (s, 1H, Ar-CHO), 5.45 (s, 1H, Ar-OH), 8.44 (s, 1H, CH (aliphatic)); 13C-NMR: 44.5, 55.7, 66.4, 114.7, 115.2, 118.1 121.4, 126.7, 127.0, 129.4, 130.8, 134.7, 135.9, 149.4, 151.2, 163.1, 187.5, 191.0; MS ES + (ToF): m/z 442 [M++1].
Compound Y8: (E)-N-(3,4-Dimethoxybenzylidene)-5-(4-formylphenyl)-3-(4-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3116 (C-H str., benzene), 1597 (C=C str., benzene), 1357 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1272 (C-N str., carbonyl), 3204 (OH str., aromatic), 1770 (CHO str., aromatic), 1222 (C=S str., aliphatic), 1645 (C=C str., aliphatic), 1017 (-NH- str., aliphatic), 2800 (C-OCH3 str., aromatic); 1H NMR: 6.21-8.21 (s, 8H, Ar-H), 1.51-3.17 (s, 3H, pyrazole), 9.72 (s, 1H, Ar-CHO), 5.55 (s, 1H, Ar-OH), 8.54 (s, 1H, CH (aliphatic)); 13C-NMR: 44.0, 56.2, 66.2, 114.4, 115.4, 118.8, 121.9, 122.5, 127.1, 129.6, 130.4, 134.4, 135.7, 149.9, 151.1, 152.2, 168.5, 187.5, 191.4; MS ES + (ToF): m/z 472 [M++1].
Compound Y9: (E)-N-(4-Hydroxybenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3091 (C-H str., benzene), 1600 (C=C str., benzene), 1358 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1277 (C-N str., carbonyl), 3514 (OH str., aromatic), 1755 (CHO str., aromatic), 1220 (C=S str., aliphatic), 1646 (C=C str., aliphatic), 1028 (-NH- str., aliphatic); 1H NMR: 6.23-7.99 (s, 8H, Ar-H), 1.81-3.58 (s, 3H, pyrazole), 9.72 (s, 1H, Ar-CHO), 5.18 (s, 1H, Ar-OH), 8.64 (s, 1H, CH (aliphatic)); 13C-NMR: 41.0, 65.2, 115.9, 116.2, 119.4, 122.2, 126.4, 127.9, 129.4, 131.1, 134.4, 135.9, 149.4, 152.1, 160.7, 162.6, 167.3, 187.9, 191.6; MS ES + (ToF): m/z 430 [M++1].
Compound Y10: (E)-N-(3-Nitrobenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3137 (C-H str., benzene), 1596 (C=C str., benzene), 1355 (C=N str., pyrazole moiety), 1167 (N-N str., pyrazole moiety), 1280 (C-N str., carbonyl), 3509 (OH str., aromatic), 1735 (CHO str., aromatic), 1215 (C=S str., aliphatic), 1689 (C=C str., aliphatic), 1003 (-NH- str., aliphatic), 1509 (C-NO2 str., aromatic);
1H NMR: 6.72-8.21 (s, 8H, Ar-H), 1.96-3.32 (s, 3H, pyrazole), 9.82 (s, 1H, Ar-CHO), 5.19 (s, 1H, Ar-OH), 8.74 (s, 1H, CH (aliphatic)); 13C-NMR: 40.9, 66.9, 155.7, 118.3, 121.2, 123.9, 124.4, 127.9, 129.0, 130.9, 134.4, 135.7, 148.2, 149.9, 152.1, 164.0, 168.9, 187.4, 191.2; MS ES + (ToF): m/z 457[M++1].
Compound Y11: (E)-N-(2-Nitrobenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3121 (C-H str., benzene), 1602 (C=C str., benzene), 1343 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1277 (C-N str., carbonyl), 3520 (OH str., aromatic), 1746 (CHO str., aromatic), 1218 (C=S str., aliphatic), 1651 (C=C str., aliphatic), 1028 (-NH- str., aliphatic), 1574 (C-NO2 str., aromatic)
1H NMR: 6.52-8.25 (s, 8H, Ar-H), 1.97-3.98 (s, 3H, pyrazole), 9.82 (s, 1H, Ar-CHO), 5.13 (s, 1H, Ar-OH), 8.18 (s, 1H, CH (aliphatic)); 13C-NMR: 40.9, 66.1, 115.3, 118.5, 121.2, 123.9, 124.5, 127.2, 129.5, 130.1, 135.2, 135.8, 148.7, 149.5, 150.6, 151.6, 163.2, 168.3, 187.2, 191.0; MS ES + (ToF): m/z 458 [M++1].
Compound Y12: (E)-N-(4-Nitrobenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3145 (C-H str., benzene), 1601 (C=C str., benzene), 1342 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1277 (C-N str., carbonyl), 3207 (OH str., aromatic), 1731 (CHO str., aromatic), 1217 (C=S str., aliphatic), 1651 (C=C str., aliphatic), 1029 (-NH- str., aliphatic), 1574 (C-NO2 str., aromatic);
1H NMR: 6.62-8.97 (s, 8H, Ar-H), 2.01-3.90 (s, 3H, pyrazole), 9.20 (s, 1H, Ar-CHO), 5.14 (s, 1H, Ar-OH), 8.19 (s, 1H, CH (aliphatic)); 13C-NMR: 40.5, 66.9, 115.5, 118.8, 121.0, 123.6, 124.4, 126.3, 127.5, 129.0, 130.7, 132.4, 134.7, 135.3, 148.9, 149.4, 151.8, 163.5, 168.0, 187.0, 190.9; MS ES + (ToF): m/z 459 [M++1].
Compound Y13: (E)-N-(3-Bromobenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3147 (C-H str., benzene), 1595 (C=C str., benzene), 1356 (C=N str., pyrazole moiety), 1167 (N-N str., pyrazole moiety), 1276 (C-N str., carbonyl), 3206 (OH str., aromatic), 1748 (CHO str., aromatic), 1217 (C=S str., aliphatic), 1650 (C=C str., aliphatic), 1017 (-NH- str., aliphatic), 617 (C-Br str., aromatic);
1H NMR: 6.23-8.21 (s, 8H, Ar-H), 1.05-3.08 (s, 3H, pyrazole), 9.02 (s, 1H, Ar-CHO), 5.16 (s, 1H, Ar-OH), 8.20 (s, 1H, CH (aliphatic)); 13C-NMR: 40.9, 66.5, 115.5, 118.4, 121.7, 123.7, 127.3, 128.5, 129.2, 130.1, 131.5, 134.6, 135.6, 149.2, 151.2, 163.0, 187.5, 191.9; MS ES + (ToF): m/z 491 [M++1].
Compound Y14: (E)-N-(4-Bromobenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3152 (C-H str., benzene), 1588 (C=C str., benzene), 1357 (C=N str., pyrazole moiety), 1167 (N-N str., pyrazole moiety), 1277 (C-N str., carbonyl), 3208 (OH str., aromatic), 1739 (CHO str., aromatic), 1218 (C=S str., aliphatic), 1651 (C=C str., aliphatic), 1028 (-NH- str., aliphatic), 619 (C-Br str., aromatic);
1H NMR: 6.24-8.22 (s, 8H, Ar-H), 1.06-3.28 (s, 3H, pyrazole), 9.31 (s, 1H, Ar-CHO), 5.17 (s, 1H, Ar-OH), 8.02 (s, 1H, CH (aliphatic)); 13C-NMR: 40.3, 66.6, 115.8, 118.0, 121.3, 123.8, 127.6, 128.3, 129.8, 130.2, 131.6, 134.5, 135.9, 149.7, 151.8, 163.6, 187.6, 191.8; MS ES + (ToF): m/z 493 [M++1].
Compound Y15: (E)-N-(2-Chlorobenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3152 (C-H str., benzene), 1591 (C=C str., benzene), 1358 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1278 (C-N str., carbonyl), 3248 (OH str., aromatic), 1747 (CHO str., aromatic), 1218 (C=S str., aliphatic), 1650 (C=C str., aliphatic), 1049 (-NH- str., aliphatic), 718 (C-Cl str., aromatic);
1H NMR: 6.34-8.32 (s, 8H, Ar-H), 1.07-3.38 (s, 3H, pyrazole), 9.32 (s, 1H, Ar-CHO), 5.18 (s, 1H, Ar-OH), 8.06 (s, 1H, CH (aliphatic)); 13C-NMR: 41.3, 65.9, 114.3, 117.5,120.9, 127.5, 129.5, 130.4, 132.9, 133.9, 134.5, 135.7, 149.4, 151.8, 163.5, 187.0, 191.3; MS ES + (ToF): m/z 446 [M++1].
Compound Y16: (E)-N-(3-Chlorobenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3147 (C-H str., benzene), 1599 (C=C str., benzene), 1357 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1278 (C-N str., carbonyl), 3215 (OH str., aromatic), 1725 (CHO str., aromatic), 1220 (C=S str., aliphatic), 1659 (C=C str., aliphatic), 1012 (-NH- str., aliphatic), 703 (C-Cl str., aromatic);
1H NMR: 6.34-8.52 (s, 8H, Ar-H), 2.06-3.29 (s, 3H, pyrazole), 9.33 (s, 1H, Ar-CHO), 5.12 (s, 1H, Ar-OH), 8.16 (s, 1H, CH (aliphatic)); 13C-NMR: 40.3, 66.9, 114.9, 117.9, 120.0, 127.6, 129.3, 130.8, 132.3, 133.5, 134.8, 135.3, 149.1, 151.0, 163.2, 187.9, 191.5; MS ES + (ToF): m/z 445 [M++1].
Compound Y17(E)-N-(4-Chlorobenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3147 (C-H str., benzene), 1599 (C=C str., benzene), 1357 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1278 (C-N str., carbonyl), 3215 (OH str., aromatic), 1725 (CHO str., aromatic), 1220 (C=S str., aliphatic), 1659 (C=C str., aliphatic), 1012 (-NH- str., aliphatic), 709 (C-Cl str., aromatic);
1H NMR: 6.44-8.24 (s, 8H, Ar-H), 1.26-3.20 (s, 3H, pyrazole), 9.71 (s, 1H, Ar-CHO), 5.37 (s, 1H, Ar-OH), 8.22 (s, 1H, CH (aliphatic)); 13C-NMR: 41.6, 65.2, 115.3, 118.5, 121.9, 127.9, 129.3, 130.8, 132.0, 133.1, 134.6, 135.4, 149.7, 151.2, 163.9, 187.7, 191.0; MS ES + (ToF): m/z 448 [M++1].
Compound Y18: (E)-N-(2,4-Dichlorobenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3059 (C-H str., benzene), 1602 (C=C str., benzene), 1357 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1279 (C-N str., carbonyl), 3203 (OH str., aromatic), 1725 (CHO str., aromatic), 1218 (C=S str., aliphatic), 1656 (C=C str., aliphatic), 1049 (-NH- str., aliphatic), 719 (C-Cl str., aromatic);
1H NMR: 6.44-7.92 (s, 8H, Ar-H), 2.16-3.28 (s, 3H, pyrazole), 8.99 (s, 1H, Ar-CHO), 5.15 (s, 1H, Ar-OH), 8.52 (s, 1H, CH (aliphatic)); 13C-NMR: 40.9, 66.0, 115.4, 118.9, 121.5, 127.3, 129.7, 130.9, 132.1, 133.6, 134.6, 135.7, 149.8, 151.9, 163.9, 187.9, 191.0; MS ES + (ToF): m/z 482 [M++1].
Compound Y19: (E)-N-(2-Hydroxy-4-methoxybenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3151 (C-H str., benzene), 1592 (C=C str., benzene), 1357 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1275 (C-N str., carbonyl), 1502 (OH str., aromatic), 1746 (CHO str., aromatic), 1223 (C=S str., aliphatic), 1650 (C=C str., aliphatic), 1031 (-NH- str., aliphatic), 2844 (OCH3 str., aromatic);
1H NMR: 5.99-8.02 (s, 8H, Ar-H), 1.66-3.08 (s, 3H, pyrazole), 9.42 (s, 1H, Ar-CHO), 5.02 (s, 1H, Ar-OH), 8.52 (s, 1H, CH (aliphatic)); 13C-NMR: 40.9, 55.3, 66.9, 102.4, 107.2, 110.9, 115.5, 118.4, 121.8, 127.6, 129.5, 130.0, 131.6, 134.7, 135.9, 149.9, 152.0, 162.2, 163.6, 164.9, 168.6, 187.0, 191.7; MS ES + (ToF): m/z 460 [M++1].
Compound Y20 :(E)-N-(4-(Dimethylamino)benzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3155 (C-H str., benzene), 1588 (C=C str., benzene), 1356 (C=N str., pyrazole moiety), 1167 (N-N str., pyrazole moiety), 1276 (C-N str., carbonyl), 3207 (OH str., aromatic), 1740 (CHO str., aromatic), 1229 (C=S str., aliphatic), 1650 (C=C str., aliphatic), 1017 (-NH- str., aliphatic), 2818 (C-(CH3)2N str., aromatic); 1H NMR: 6.74-8.45 (s, 8H, Ar-H), 1.72-3.42 (s, 3H, pyrazole), 9.26 (s, 1H, Ar-CHO), 5.37 (s, 1H, Ar-OH), 8.42 (s, 1H, CH (aliphatic)); 13C-NMR: 40.3, 66.3, 114.5, 115.9, 118.5, 121.9, 123.4, 127.8, 129.5, 130.1, 134.7, 135.7, 148.9, 150.9, 163.0, 168.3, 187.1, 191.9; MS ES + (ToF): m/z 457 [M++1].
Compound Y21: (E)-N-(4-(Diethylamino)benzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3139 (C-H str., benzene), 1586 (C=C str., benzene), 1352 (C=N str., pyrazole moiety), 1170 (N-N str., pyrazole moiety), 1274 (C-N str., carbonyl), 3201 (OH str., aromatic), 1721 (CHO str., aromatic), 1245 (C=S str., aliphatic), 1654 (C=C str., aliphatic), 1032 (-NH- str., aliphatic); 1H NMR: 6.92-8.42 (s, 8H, Ar-H), 1.72-3.25 (s, 3H, pyrazole), 9.27 (s, 1H, Ar-CHO), 5.27 (s, 1H, Ar-OH), 8.47 (s, 1H, CH (aliphatic)); 13C-NMR: 13.5,91.2, 40.6, 44.7, 66.6, 114.3, 115.9, 118.4, 121.1, 123.5, 127.3, 129.4, 130.2, 13.4, 135.2, 139.4, 148.5, 151.9, 163.6, 187.0, 191.2;MS ES + (ToF): m/z 483[M++1].
Compound Y22: (E)-N-((E)-4-(E)-Styrylbenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3058 (C-H str., benzene), 1598 (C=C str., benzene), 1354 (C=N str., pyrazole moiety), 1169 (N-N str., pyrazole moiety), 1278 (C-N str., carbonyl), 3202 (OH str., aromatic), 1763 (CHO str., aromatic), 1169 (C=S str., aliphatic), 1693 (C=C str., aliphatic), 1031(-NH- str., aliphatic); 1H NMR: 7.24-8.47 (s, 8H, Ar-H), 1.92-3.27 (s, 3H, pyrazole), 9.54 (s, 1H, Ar-CHO), 5.22 (s, 1H, Ar-OH), 8.23 (s, 1H, CH (aliphatic)); 13C-NMR: 44.3, 66.2, 115.3, 118.2, 119.5, 121.1, 126.3, 127.5, 128.5, 129.6, 130.0, 134.6, 135.9, 140.7, 149.9, 151.0, 163.7, 168.0, 187.7, 191.9; MS ES + (ToF): m/z 516 [M++1].
Compound Y23: (E)-N-(3-Methylbenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3153 (C-H str., benzene), 1589 (C=C str., benzene), 1357 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1277 (C-N str., carbonyl), 3202 (OH str., aromatic), 1721 (CHO str., aromatic), 1237 (C=S str., aliphatic), 1641 (C=C str., aliphatic), 1016 (-NH- str., aliphatic), 3011 (CH3 str., aromatic);
1H NMR: 6.28-8.86 (s, 8H, Ar-H), 1.97-3.25 (s, 3H, pyrazole), 9.24 (s, 1H, Ar-CHO), 5.32 (s, 1H, Ar-OH), 8.13 (s, 1H, CH (aliphatic)); 13C-NMR: 24.5, 44.0, 65.9, 116.2, 119.9, 121.0, 127.4, 129.8, 129.9,130.8, 134.5, 135.9, 140.7, 149.5, 151.9, 163.7, 168.2, 187.7, 191.3; MS ES + (ToF): m/z 426 [M++1].
Compound Y24: (E)-N-(3,4,5-Trimethoxybenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3154 (C-H str., benzene), 1595 (C=C str., benzene), 1356 (C=N str., pyrazole moiety), 1168 (N-N str., pyrazole moiety), 1269 (C-N str., carbonyl), 3206 (OH str., aromatic), 1722 (CHO str., aromatic), 1238 (C=S str., aliphatic), 1265 (C=C str., aliphatic), 1017 (-NH- str., aliphatic), 2859 (OCH3 str., aromatic);
1H NMR: 6.65-8.87 (s, 8H, Ar-H), 1.98-3.24 (s, 3H, pyrazole), 9.52 (s, 1H, Ar-CHO), 5.66 (s, 1H, Ar-OH), 8.21 (s, 1H, CH (aliphatic)); 13C-NMR: 40.3, 56.6, 66.3, 106.6, 115.9, 118.5, 121.3, 127.5, 128.9, 129.1, 130.9, 134.2, 135.7, 149.4, 150.3, 151.9, 164.1, 168.3, 187.7, 192.0; MS ES + (ToF): m/z 504 [M++1].
Compound Y25: (E)-N-(2,6-Dichlorobenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3128 (C-H str., benzene), 1592 (C=C str., benzene), 1356 (C=N str., pyrazole moiety), 1169 (N-N str., pyrazole moiety), 1274 (C-N str., carbonyl), 3505 (OH str., aromatic), 1739 (CHO str., aromatic), 1216 (C=S str., aliphatic), 1641 (C=C str., aliphatic), 1016 (-NH- str., aliphatic), 709 (C-Cl str., aromatic);
1H NMR: 6.29-8.18 (s, 8H, Ar-H), 1.29-3.27 (s, 3H, pyrazole), 9.28 (s, 1H, Ar-CHO),5.26 (s, 1H, Ar-OH), 8.04(s, 1H, CH (aliphatic)); 13C-NMR: 40.1, 66.4, 115.0, 118.2, 121.8, 127.5, 129.7, 130.3, 131.9, 133.9, 134.9, 135.4, 149.3, 151.8, 163.7, 168.9, 187.0, 191.1; MS ES + (ToF): m/z 481 [M++1].
Compound Y26: (E)-N-(3-Hydroxybenzylidene)-5-(4-formylphenyl)-3-(3-hydroxyphenyl)-4,5-dihydropyrazole-1-carbothioamide, IR: 3024 (C-H str., benzene), 1602 (C=C str., benzene), 1357 (C=N str., pyrazole moiety), 1167 (N-N str., pyrazole moiety), 1278 (C-N str., carbonyl), 3505 (OH str., aromatic), 1727 (CHO str., aromatic), 1220 (C=S str., aliphatic), 1649 (C=C str., aliphatic), 1031 (-NH-str., aliphatic); 1H-NMR: 6.92-8.91 (s, 8H, Ar-H), 1.21-3.77 (s, 3H, pyrazole), 9.87 (s, 1H, Ar-CHO), 5.26 (s, 1H, Ar-OH), 8.22 (s, 1H, CH (aliphatic)); 13C-NMR: 40.4, 66.4, 115.3, 118.8, 121.9, 127.9, 129.5, 130.5, 134.5, 135.9, 149.5, 151.3, 158.3, 163.2, 168.7, 187.5, 191.9; MS ES + (ToF): m/z 430 [M++1].