TRIAZOLE AND HYDRAZIDE-HYDRAZONE DERIVATIVE HYBRID MOLECULES WITH ANTITUMORAL EFFECT AND THE SYNTHESIS METHODS OF THESE MOLECULES

Technical Field of the Invention

The Invention relates to hydrazide and hydrazlde-hydrazone derivative molecules with antitumoral effect, bearing 1 ,2,4- triazole rings, which are effective on the MCF-7 breast cancer cell line and on cyclooxygenase-1 (COX-1 ), especially having a selective Inhibition activity on the cyclooxygenase-2 (COX-2) enzyme, and synthesis methods of these molecules.

State of the Art

Inflammation Is a response of the Immune system to protect the body against various diseases or Injuries. When tissue homoeostasls Is disrupted, macrophages and mast cells release bioactive stimuli such as cytokines, chemoklnes, ROS and histamine, resulting In leukocyte mobilisation and leakage at the site of Injury assodated with the Inflammatory process [1j. Continuation of these responses for a long time causes chronic Inflammation and results In many diseases, especially cancer [2, 3j. Clinical and epidemiological studies show a strong relationship between chronic Infection, Inflammation and cancer. Acute and chronic Inflammation can cause many diseases such as atherosclerosis, heart disease, cancer, arthritis, metabolic syndrome, high blood pressure, Parkinson's disease, Alzheimer's disease, asthma, Inflammatory bowel disease and diabetes.

In the state of the art, non-steroidal anti-inflammatory drugs (NSAIDs) are used to Inhibit Inflammation. Cyclooxygenase (COX), which is one of the enzymes targeted by these drugs and consists of three isoforms, is an enzyme that is targeted in pain relief and inflammation treatment. Cyclooxygenase-2 (COX-2), one of the COX Isoforms, Is a prostaglandin endoperoxide synthase 2 enzyme responsible for the formation of prostanoids that contribute to the modulation of multiple procarclnogenlc effects. It has been shown that this enzyme Is overexpressed In many types of cancer and Is used as a marker. As a result of overexpression of COX-2, carcinogenesis Is promoted, Increasing the recurrence rate and Invasion of cancer, decreasing the survival rate, and making cancer cells resistant to radiotherapy and chemotherapy. COX-1 , on the other hand, Is an enzyme responsible for the synthesis of PGE2 and PGI2, which has cytoprotectlve effects on various aspects of gastric function, such as Increasing bicarbonate and mucus secretion, decreasing gastric add and pepsin secretion, and ensuring adequate blood flow to the mucosa[4j. Examples of COX Inhibitors In the state of the art Include celecoxlb, Indomethacin, metamlzole, aspirin and aminopyrin.

Although non-steroidal anti-inflammatory drugs have been developed In the state of the art for the Inhibition of Inflammation, which Is of great Importance In the treatment or prevention of cancer, Inflammation Inhibition alone Is not effective enough In the treatment of cancer. Although significant progress has been made In some types of cancer In the last 30 years In the efforts to develop effective treatment protocols against cancer, when the general success rate Is considered, It Is seen that the desired level of progress In terms of five-year survival has not been achieved. This situation and the Increasing Incidence of cancer worldwide necessitates the creation of new approaches to cancer approach and treatment

In the state of the art, the most common type of cancer diagnosed In women Is breast cancer, and It ranks second among the causes of death In women worldwlde[5]. In the state of the art, scientific studies for the treatment of breast cancer are carried out through studies on the MCF-7 breast cancer cell line. It Is possible to find studies on the state of the art to measure the effects of the 1 ,2,4-triazole ring, which has many pharmacological activities, on the MCF-7 breast cancer cell line. In fact, there are studies In the literature to measure the effectiveness of molecules containing hydrazide and hydrazone derivatives on the MCF-7 breast cancer cell line.

In a study conducted by Yuril G. SAMELIUK et al., the activities of 1 ,2,4-triazole derivatives In medldne and pharmacy were measured. In said scientific study, It Is stated that the molecules subject to the study, which contain a 1 ,2,4-triazole ring, have anticancer properties, and their effectiveness on MCF-7 cells Is also mentioned[6].

1 Although there are non-steroidal anti-inflammatory drugs (NSAIDs) that play a very important role in cancer treatment and are used to inhibit inflammation, which is a response created by the immune system to protect the body against various diseases or injuries, in the state of the art, said drugs do not have sufficient activity, especially on breast cancer, which is one of the deadliest types of cancer today. Although, in the state of the art, studies are being carried out to measure the effects of the 1 ,2,4-triazole ring on the MCF-7 breast cancer cell line, the molecules synthesized in these studies do not have the ability to prevent the overexpression of COX enzymes, which are targets for pain relief and inflammation treatment. In other words, even if said molecules are effective on the MCF-7 breast cancer cell line, since they cannot prevent the overexpression of COX enzymes, carcinogenesis will be encouraged, the recurrence rate and invasion of cancer will be increased, the survival rates of cancer patients will decrease, and worst of all, cancer cells will become resistant to radiotherapy and chemotherapy.

Due to problems such as limitations and inadequacies of the drugs or synthesized molecules in the state of the art and especially used in the treatment of breast cancer, 1 ,2,4-triazole derivative molecules developed in the state of the art for use in the treatment of breast cancer being unable to prevent the overexpression of COX enzymes even though they are effective on the MCF-7 breast cancer cell line, and therefore promoting carcinogenesis, increasing the recurrence rate and invasion of cancer, thus decreasing the survival rates of cancer patients, and causing fatal consequences such as cancer cells becoming resistant to radiotherapy and chemotherapy, a new molecule that is both effective on the MCF-7 breast cancer cell line and has an inhibitory activity on COX-1 and COX-2 isoenzymes is needed for use in cancer treatment

Brief Description and Aims of the Invention

In the inventio, hydrazide and hydrazide-hydrazone derivative molecules with antitumoral effect, bearing 1 ,2,4-triazole rings, which are effective on the MCF-7 breast cancer cell line and on cyclooxygenase-1 (COX-1 ), especially having a selective inhibition activity on the cyclooxygenase-2 (COX-2) enzyme, and synthesis methods of these molecules are described. The general structure of the molecules of the invention is shown by Formula X.

An aim of the invention is to provide effective molecules for use in cancer treatment, especially on the MCF-7 breast cancer cell line. The determination of said molecules is achieved by the synthesis method of hydrazide and hydrazide-hydrazone derivative antitumoral effective molecules bearing a 1 ,2,4-triazole ring. The mechanism of action of the molecules of the invention on the MCF-7 breast cancer cell line is understood by measuring the inhibition of substances applied at certain times and at different concentrations on the MCF-7 cell line.

Another aim of the invention is to provide molecules that have an activity on the COX-1 isoenzyme and especially a selective inhibition activity on the cyclooxygenase-2 (COX-2) enzyme. The determination of said molecules is achieved by the synthesis method of hydrazide and hydrazide-hydrazone derivative antitumoral effective molecules bearing a 1 ,2,4- triazole ring.

In the invention, molecules that are effective both on the MCF-7 breast cancer cell line and cyclooxygenase-1 (COX-1 ) and have a selective inhibition activity especially on the cyclooxygenase-2 (COX-2) enzyme are presented. The determination of said molecules is achieved by the synthesis method of hydrazide and hydrazide-hydrazone derivative antitumoral effective molecules bearing a 1 ,2,4-triazole ring. The aim of the invention is to reveal molecules that increase the survival rates of cancer patients and prevent cancer cells from becoming resistant to radiotherapy and chemotherapy. The determination of said molecules is achieved by the synthesis method of hydrazide and hydrazide-hydrazone derivative antitumoral effective molecules bearing a 1 ,2,4-triazole ring. The molecules that are the subject of the invention provide inhibition of COX-2, which is one of the COX isoforms and whose overexpression promotes tumor formation, that is, carcinogenesis, increases the rate of cancer recurrence and invasion, reduces the survival rate of cancer patients, and makes cancer cells resistant to radiotherapy and chemotherapy.

Description of Drawings

Figure 1. Comparative column chart showing the effect of 24-hour application of the molecules of the invention and cisplatin and osimertinib used as standards on the viability of MCF-7 cells

Figure 2. Comparative column chart showing the effect of 24-hour application of the molecules of the invention and the substances used as standards, cisplatin and osimertinib, on the viability of HEK293 cells

Figure 3. Fourier Transform Infrared Spectroscopy (FTI R) spectrum of Formula 1

Figure 4. ¹ H-NMR spectrum of formula 1 Figure 34. ¹³ C-NMR (APT) spectrum of formula 2f

Figure 5. ¹³ C-NMR (APT) spectrum of formula 1 Figure 35. ESI-HRMS spectrum of formula 2f (positive)

Figure 6. ESI-HRMS spectrum of formula 1 (negative) Figure 36. ESI-HRMS spectrum of formula 2f (negative)

Figure 7. FTI R spectrum of formula 2a Figure 37. FTIR spectrum of formula 2g

Figure 8. ¹ H-NMR spectrum of formula 2a Figure 38. ¹ H-NMR spectrum of formula 2g

Figure 9. ¹³ C-NMR (APT) spectrum of formula 2a Figure 39. ¹³ C-NMR (APT) spectrum of formula 2g

Figure 10. ESI-HRMS spectrum of formula 2a (positive) Figure 40. ESI-HRMS spectrum of formula 2g (positive)

Figure 11. ESI-HRMS spectrum of formula 2a (negative) Figure 41. ESI-HRMS spectrum of formula 2g (negative)

Figure 12. FTIR spectrum of formula 2b Figure 42. FTIR spectrum of formula 2h

Figure 13. ¹ H-NMR spectrum of formula 2b Figure 43. ¹ H-NMR spectrum of formula 2h

Figure 14. ¹³ C-NMR (APT) spectrum of formula 2b Figure 44. ¹³ C-NMR (APT) spectrum of formula 2h

Figure 15. ESI-HRMS spectrum of formula 2b (positive) Figure 45. ESI-HRMS spectrum of formula 2h (positive)

Figure 16. ESI-HRMS spectrum of formula 2b (negative) Figure 46. ESI-HRMS spectrum of formula 2h (negative)

Figure 17. FTIR spectrum of formula 2c Figure 47. FTIR spectrum of formula 2i

Figure 18. ¹ H-NMR spectrum of formula 2c Figure 48. ¹ H-NMR spectrum of formula 2i

Figure 19. ¹³ C-NMR (APT) spectrum of formula 2c Figure 49. ¹³ C-NMR (APT) spectrum of formula 2i

Figure 20. ESI-HRMS spectrum of formula 2c (positive) Figure 50. ESI-HRMS spectrum of formula 2i (positive)

Figure 21. ESI-HRMS spectrum of formula 2c (negative) Figure 51. ESI-HRMS spectrum of formula 2i (negative)

Figure 22. FTIR spectrum of formula 2d Figure 52. FTIR spectrum of formula 2j

Figure 23. ¹ H-NMR spectrum of formula 2d Figure 53. ¹ H-NMR spectrum of formula 2j

Figure 24. ¹³ C-NMR (APT) spectrum of formula 2d Figure 54. ¹³ C-NMR (APT) spectrum of formula 2j

Figure 25. ESI-HRMS spectrum of formula 2d (positive) Figure 55. ESI-HRMS spectrum of formula 2j (positive)

Figure 26. ESI-HRMS spectrum of formula 2d (negative) Figure 56. ESI-HRMS spectrum of formula 2j (negative)

Figure 27. FTIR spectrum of formula 2e Figure 57. FTIR spectrum of formula 2k

Figure 28. ¹ H-NMR spectrum of formula 2e Figure 58. ¹ H-NMR spectrum of formula 2k

Figure 29. ¹³ C-NMR (APT) spectrum of formula 2e Figure 59. ¹³ C-NMR (APT) spectrum of formula 2k

Figure 30. ESI-HRMS spectrum of formula 2e (positive) Figure 60. ESI-HRMS spectrum of formula 2k (positive)

Figure 31. ESI-HRMS spectrum of formula 2e (negative) Figure 61. ESI-HRMS spectrum of formula 2k (negative)

Figure 32. FTIR spectrum of formula 2f Figure 62. FTIR spectrum of formula 2g

Figure 33. ¹ H-NMR spectrum of formula 2f Figure 63. ¹ H-NMR spectrum of formula 2g

Figure 64. ¹³ C-NMR (APT) spectrum of formula 2I Figure 83. ¹ H-NMR spectrum of formula 2p

Figure 65. ESI-HRMS spectrum of formula 2I (positive) Figure 84. ¹³ C-NMR (APT) spectrum of formula 2p

Figure 66. ESI-HRMS spectrum of formula 2g (negative) Figure 85. ESI-HRMS spectrum of formula 2p (positive)

Figure 67. FTIR spectrum of formula 2m Figure 86. ESI-HRMS spectrum of formula 2p (negative) Figure 68. ¹ H-NMR spectrum of formula 2m Figure 87. FTIR spectrum of formula 2r

Figure 69. ¹³ C-NMR (APT) spectrum of formula 2m Figure 88. ¹ H-NMR spectrum of formula 2r

Figure 70. ESI-HRMS spectrum of formula 2m (positive) Figure 89. ¹³ C-NMR (APT) spectrum of formula 2r

Figure 71. ESI-HRMS spectrum of formula 2m (negative) Figure 90. ESI-HRMS spectrum of formula 2r (positive)

Figure 72. FTIR spectrum of formula 2n Figure 91. ESI-HRMS spectrum of formula 2r (negative)

Figure 73. ¹ H-NMR spectrum of formula 2n Figure 92. FTIR spectrum of formula 2s

Figure 74. ¹³ C-NMR (APT) spectrum of formula 2n Figure 93. ¹ H-NMR spectrum of formula 2s

Figure 75. ESI-HRMS spectrum of formula 2n (positive) Figure 94. ¹³ C-NMR (APT) spectrum of formula 2s

Figure 76. ESI-HRMS spectrum of formula 2n (negative) Figure 95. ESI-HRMS spectrum of formula 2s (positive)

Figure 77. FTIR spectrum of formula 2o Figure 96. ESI-HRMS spectrum of formula 2s (negative)

Figure 78. ¹ H-NMR spectrum of formula 2o Figure 97. FTIR spectrum of formula 2t

Figure 79. ¹³ C-NMR (APT) spectrum of formula 2o Figure 98. ¹ H-NMR spectrum of formula 2t

Figure 80. ESI-HRMS spectrum of formula 2o (positive) Figure 99. ¹³ C-NMR (APT) spectrum of formula 2t

Figure 81. ESI-HRMS spectrum of formula 2o (negative) Figure 100. ESI-HRMS spectrum of formula 2t (positive)

Figure 82. FTIR spectrum of formula 2p Figure 101. ESI-HRMS spectrum of formula 2t (negative)

Detailed Description of the Invention

The invention relates to hydrazide and hydrazide-hydrazone derivative molecules with antitumoral effect, bearing 1 ,2,4- triazole rings, which are effective on the MCF-7 breast cancer cell line and on cyclooxygenase-1 (COX-1 ), especially having a selective inhibition activity on the cyclooxygenase-2 (COX-2) enzyme, and synthesis methods of these molecules. The general structure of the molecules of the invention is shown by Formula X.

Formula X wherein R is selected from the formulas below: In said Formula X, If the R ¹ group is R ¹ = -HN - NH2, the obtained 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3- yl]sulfanyl]acetohydrazide molecule is represented by Formula 1.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1,2,4-triazol-3- yl]sulfanyl]acetohydrazide (Formula 1)

Formula 1

It was obtained as a white powder with a yield of 60-95% and its melting temperature is in the range of 166-168oC.

Fourier Transform Infrared Spectroscopy (FTIR) v (cm ^-1 ): 3317, 3257 (N-H stretching band); 3152, 3043 (ar C-H stretching band); 2900, 2850 (aldehyde C-H asymmetric and symmetric stretching band), 1679 (C=O stretching band); 1610 (hydrazone C=N stretching band), 1506 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands); 1429, 1398 (aldehyde C-H asymmetric and symmetric stretching band); 1215 (C-S stretching band), 1205 (aromatic C-F stretching band), 837 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 9,37 (s, 1 H, CONH); 7,77 (d, J: 1 ,6 Hz, 1 H, furan C ₅ -H); 7,63-7,59 (m, 2H, FPhC2, ₆ -H); 7,47 (t, J: 8.65 Hz, 2H, FPhC ₃ ,5-H); 6,54 (dd, J: 3.5; 1.6 Hz, 1 H, furan C ₄ -H); 6,24 (d, J: 3.5 Hz, 1 H, furan C ₃ -H); 4,32 (s, 2H, NH ₂ ); 3,88 (s, 2H, SCH ₂ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 166.43, 163.22 (d, J: 252 Hz), 151.8, 148.01 , 145.34, 141.40, 130.71 (d, J: 9,5 Hz), 130.06 (d, J: 3 Hz), 1 17.47 (d, J: 22 Hz), 1 12.15, 11 1.8, 34.9

ESI-HRMS: mass: m/z 333.0695 calculated [M+K]+; m/z 372.0332, found [M+K]+: m/z 372.0943; calculated for Ci4Hi ₂ FN ₅ O ₂ S: 50.44%C 3.63%H 21 .01 %N, Found: 50.31 %C 3.82%H 21 ,03%N.

In said Formula X, if the R ¹ group is R ¹ = , molecule derivatives represented by formula 2a-t are obtained. Molecules represented by formula 2a-t are 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4- triazol-3-yl]sulfanyl]-N'-[(substituted phenyl)methylidene]acetohydrazide derivatives.

Formula 2a-t In the Formulas 2a-t, the RI I group is selected from the formulas below;

Formula r Formula 2a-t molecules are as follows:

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(phenyl)methylidene]acetohyd razide (Formula 2a)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(2,3- dihydroxyphenyl)methylidine]acetohydrazide (Formula 2b)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(2-hydroxy-4- diethylaminophenyl)methylidene]acetohydrazide (Formula 2c)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(2-hydroxy-3- methoxyphenyl)methylidene]acetohydrazide (Formula 2d)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- ethylphenyl)methylidene]acetohydrazide (Formula 2e)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(2,4- dinitrophenyl)methylidine]acetohydrazide (Formula 2f)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(3,4,5- trimethoxyphenyl)methylidene]acetohydrazide (Formula 2g)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- nitrophenyl)methylidene]acetohydrazide (Formula 2h)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- methylphenyl)methylidene]acetohydrazide (Formula 2i)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(3,4- dimethoxyphenyl)methylidine]acetohydrazide (Formula 2j)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4-methoxy)methylidene]aceto hydrazide (Formula 2k)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- cyanophenyl)methylidene]acetohydrazide (Formula 2I)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- hydroxyphenyl)methylidene]acetohydrazide (Formula 2m)

. 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(3- hydroxyphenyl)methylidene]acetohydrazide (Formula 2n)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- chlorophenyl)methylidene]acetohydrazide (Formula 2o)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- fluorophenyl)methylidene]acetohydrazide (Formula 2p)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- bromophenyl)methylidene]acetohydrazide (Formula 2r)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(2- hydroxyphenyl)methylidene]acetohydrazide (Formula 2s)

• 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- dimethylaminophenyl)methylidene]acetohydrazide (Formula 2t)

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1,2,4-triazol-3- yl]sulfanyl]-N'-[(phenyl)methylidene]acetohydrazide (Formula 2a)

When RII=-H in Formula 2a-t, the molecule shown in Formula 2a is obtained.

Formula 2a

It was obtained as a white powder with a yield of 60-95% and its melting temperature is in the range of 1 19-121 °C.

FTIR v (cm ¹ ): 3419 (N-H stretching band); 3177, 3086 (aromatic C-H stretching band); 2964 (aldehyde C-H asymmetric and symmetric stretching band), 1670 (amide C=O stretching band); 1600 (hydrazone C=N stretching band), 1509 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands); 1403, 1383 (aldehyde C-H asymmetric and symmetric stretching band), 1223 (C-S stretching band), 1202 (aromatic C-F stretching band), 841 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d6) 5 (ppm): 1 1 ,73 and 11 ,68 (2s, 1 H, CONH); 8,19 and 8,01 (2s, 1 H, N=CH); 7,77 (d, J: 1 ,8 Hz, 1 H, furan C ₅ -H); 7,49-7,42 (m, 9H, aromatic C-H); 6,53 (dd, J: 3,4; 1 ,8 Hz, 1 H, furan C ₄ -H); 6,23 (d, J: 3,4 Hz, 1 H, furan C ₃ -H); 4,48 and 4,07 (2s, 2H, SCH ₂ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-de) 6 (ppm): 168,90 and 163,74 (C=O); 163,21 (d, J: 248 H); 151 ,95; 147,85; 147,65 and 144,41 (C=N); 145,32; 141 ,38; 134,36; 130,69 (d, J: 9,17 Hz); 130,47; 130,13 (d, J: 3 Hz); 129,28; 127,33; 1 17,46 (d, J: 23 Hz); 1 12,13; 1 1 1 ,80; 35,68 and 35,61

- ESI-HRMS: mass: m/z 421.1008, calculated [M+H] ⁺ : m/z 422.1087 found [M+H] ⁺ : m/z 422.1091 , calculated [M- H] ⁺ : m/z 420.0930, found [M-H] ⁺ : m/z 420.0946; calculated for C21H16FN5O2S.O.5H2O: 58.59%C 3.98%H 16.27%N, Found: 58.17%C 3.909%H 15.79%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(2,3- dihydroxyphenyl)methylidine]acetohydrazide (Formula 2b)

When RII = in Formula 2a-t, the molecule shown in Formula 2b is obtained.

Formula 2b It was obtained as a white powder substance with a yield of 60-95%. Its melting temperature is in the range of 190-192.5°C.

FTIR v (cm ¹ ): 3509 (O-H stretching band), 3169 (N-H stretching band), 3028 (ar C-H stretching band), 2970, 2928,2855 (aldehyde C-H asymmetric and symmetric stretching band), 1682 (amide C=O stretching band), 1578 (hydrazone C=N stretching band), 1509 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1449, 1359 (aldehyde C-H asymmetric and symmetric stretching band), 1225 (C-S stretching band), 1 158 (aromatic C-F stretching band), 842 (1 ,4-disubstituted aromatic C-H stretching band);

¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1 ,98 and 1 1 ,62 (2s, 1 H, CONH); 10,75 (br.s., 1 H, OH); 9,56 (br.s., 1 H, OH); 8,38 and 8,31 (2s, 1 H, N=CH); 7,78 (d, J: 1 .8 Hz, 1 H, furan C ₅ -H); 7,64-7,60 (m, 2H, FPhC ₂ ,6-H); 7,47 (td, J: 8,73; 2,09 Hz; 2H, FPhC ₃ ,5-H); 7.1 (d, J: 7.8, 1 H aromatic C ₅ -H); 6.85 (d, J: 7.8, 1 H, aromatic C ₄ -H); 6.68 (t, J: 7.8 Hz, 1 H, aromatic C ₅ -H); 6.54 (dd J: 3.3; 1 .8 Hz, 1 H, furan C ₄ -H); 6.24 (d, J: 3.3 Hz, 1 H, furan C ₃ -H); 4.47 and 4.09 (2s, 2H, SCH ₂ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168,46 and 163,60 (C=O); 163,2 (d, J: 248 Hz); 151 ,99; 148,57 and 142,78 (C=N); 147,83; 146,44; 146,03; 145,38; 141 ,37; 130,69 (d, J: 9,04 Hz); 130,1 1 (d, J: 2,76 Hz); 120,27; 1 19,67; 1 19,17; 1 17,92; 1 17,18; 1 17,49 (d, J: 23,29 Hz); 1 12,16; 11 1 ,80; 35,74 and 35,35

ESI-HRMS: mass: m/z 453.0907, calculated [M+H] ⁺ : m/z 454.0985, found [M+H] ⁺ : m/z 454.0990, calculated [M- H] ⁺ : m/z 452.0828, found [M-H] ⁺ : m/z 452.0845; calculated for C ₂₁ H ₁₆ FN ₅ O ₄ S.0.5H2O: 54.54%C 3.71 %H 15.14%N, Found: 54.76%C 3.547%H 15.21 %N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(2-hydroxy-4- diethylaminophenyl)methylidene]acetohydrazide (Formula 2c)

When Formula 2a-t, the molecule shown in Formula 2c is obtained.

Formula 2c

It was obtained as a white powder with a yield of 60-95% and its melting temperature is in the range of 247-249°C.

- FTIR v (cm- ¹ ): 3183 (N-H stretching band), 3051 (ar C-H stretching band), 2976, 2932, 2870 (aldehyde C-H asymmetric and symmetric stretching band), 1664 (amide C=O stretching band), 1600 (hydrazone C=N stretching band), 1509 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1356 (aldehyde C-H asymmetric and symmetric stretching band) 1224 (C-S stretching band), 1 197 (aromatic C-F stretching band), 848 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 6 (ppm): 1 1 ,66 and 1 1 ,33 (2s, 1 H, CONH); 1 1 ,10 (s, 1 H, OH); 8,18 and 8,1 1 (2s, 2H, N=CH); 7,77 (d, J = 1 ,8 Hz, 1 H, furan C ₅ -H); 7,63-7,60 (m, 2H, FPhC ₂ ,6-H); 7,47 (t, J: 8,7 Hz, 2H, FPhC ₃ , ₅ - H); 7,21 (d, J: 8,92 Hz, 1 H, aromatic C ₆ -H); 6,54 (dd, J = 3,54; 1 ,8 Hz, 1 H, furan C ₄ -H); 6,22-6,27 (m, 2H, furan C ₃ -H and aromatic C5-H), 6,10 (d, J: 2,4 Hz, 1 H, aromatic C ₃ -H); 4,41 and 4,01 (2s, 2H, SCH ₂ ); 3,35 (with N- CHgCHs, DMSO peaks); 1 ,1 (t, J: 7,0 Hz, 6H, 2N-CH2CH ₃ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 167,60 and 162,70 (C=O); 163,22 (d, J: 248 Hz); 160,02; 151 ,69; 149,62 and 144,22 (C=N); 150,69; 147,93; 145,38; 141 ,33; 131 ,93; 130,71 (d, J: 9,36 Hz); 130,01 (d, J: 3,2 Hz); 1 17,48 (d, J: 23 Hz); 1 12,15; 1 1 1 ,85; 106,64; 104,14; 97,84; 44,26; 35,80 and 35,35; 12,99

- ESI-HRMS: mass: m/z 508.1692, calculated [M+H] ⁺ : m/z 509.1771 , found [M+H] ⁺ : m/z 509.1781 , calculated [M- H] ⁺ : m/z 507.1614, found [M-H] ⁺ : m/z 507.1629; calculated for C25H2 ₅ FN ₆ O ₃ S: 59.04%C 4.95%H 16.52%N, Found: 58.70%C 5.095%H 16.49%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(2-hydroxy-3- methoxyphenyl)methylidene]acetohydrazide (Formula 2d)

When R" = in Formula 2a-t, the molecule shown in Formula 2d is obtained.

It was obtained as a white powder with a yield of 60-95% and its melting temperature is in the range of 21 1 -214°C.

FTIR v (cm ¹ ): 3534 (O-H stretching band), 3162 (N-H stretching band), 3063 (ar C-H stretching band), 2974, 2930, 2838 (aldehyde C-H asymmetric and symmetric stretching band), 1674 (amide C=O stretching band), 1605 (hydrazone C=N stretching band), 1511 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1456, 1382 (aldehyde C-H asymmetric and symmetric stretching band), 1226 (C-S stretching band), 1 184 (aromatic C-F stretching band), 849 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1 .92 and 1 1 .60 (2s, 1 H, CONH); 10.57 (s, 1 H, OH); 8.40 and 8.32 (2s, 2H, N=CH); 7.77 (d, J: 1.7 Hz, 1 H, furan C ₅ -H); 7.63-7.60 (m, 2H, FPhC ₂ ,6-H); 7.47 (td, 2H, J: 8.81 , 2.15 Hz, FPhC3,5-H); 7.15 (dd, J: 8; 1 .3 Hz, aromatic C6-H); 7.03 (dd, J: 8, 1 .3 Hz, aromatic C4-H); 6.85 (t, J: 8 Hz, aromatic C5-H); 6.51 (dd, J: 3.6, 1 .7 Hz, 1 H, furan C ₄ -H); 6.21 (d, J: 3.6 Hz, 1 H, furan C3-H); 4.44 and 4.05 (2s, 2H, SCH ₂ ); 3.79 (s, 3H, OCH3)

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168,57 and 163,56 (C=O); 163,21 (d, J: 248 Hz); 151 ,97; 151 ,65; 148,46; 147,82; 147,54 and 141 ,66 (C=N); 145,32; 141 ,31 ; 130,71 (d, J: 9,13 Hz); 130,14 (d, J: 2,76 Hz); 120,87; 1 19,54; 1 19,36; 1 17,46 (d, J: 23 Hz); 114,25; 1 12,15; 1 1 1 ,87; 56,31 ; 35,69 and 35,3

- ESI-HRMS: mass: m/z 467.1063, calculated [M+H] ⁺ : m/z 468.1 141 , found [M+H] ⁺ : m/z 468.1 148, calculated [M- H] ⁺ : m/z 466.0985, found [M-H] ⁺ : m/z 466.1000; calculated for C22Hi8FN ₅ O4S.0,5 C2H5-OH 56.32%C 4.32%H 14.28%N, Found: 56.00%C 4.508%H 14.24%N

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4-ethylphenyl)methylidene]a cetohydrazide (Formula 2e)

When

Formula 2e

It was obtained as a white powder with a yield of 60-95% and its melting temperature is in the range of 190-192.5°C.

- FTIR v (cm- ¹ ): 3172, 3113 (N-H stretching band), 3077 (ar C-H stretching band), 2962, 2932, 2872 (aldehyde C- H asymmetric and symmetric stretching band), 1672 (amide C=O stretching band), 1617 (hydrazone C=N stretching band), 1508(aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1473, 1430, 1383 (aldehyde C-H asymmetric and symmetric stretch band), 1222 (C-S stretch band), 1 154 (aromatic C-F stretch band), 832 (1 ,4-disubstituted aromatic C-H stretch band)

- ¹ H-NMR (500 MHz) (DMSO-cfs) 5 (ppm): 1 1 ,62 (s, 1 H, CONH); 8,15 and 7,98 (2s, 1 H, N=CH); 7.78 (d, J: 1.7 Hz, 1 H, furan C ₅ -H); 7.63-7.59 (m, 2H, FPhC ₂ ,6-H); 7,57 (d, J: 8,1 1 Hz, 2H, aromatic C ₂ , ₆ -H); 7.47 (td, 2H, J: 8.76, 2.4 Hz, FPhC ₃ ,5-H); 7,28 (d, J: 8,1 1 Hz, 2H, aromatic C ₃ ,5-H); 6.54 (dd, J: 3.6, 1 .7 Hz, 1 H, furan C ₄ -H); 6.22 (d, J: 3.6 Hz, 1 H, furan C ₃ -H); 4,46 and 4,06 (2s, 2H, SCH ₂ ); 2,65 (with Ar-CH ₂ -CH ₃ , DMSO peaks); 1 ,19 (t, J: 7.58 Hz, 3H, Ar-CH ₂ -CH ₃ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168,77 and 163,56 (C=O); 163,2 (d, J: 248 Hz); 151 ,95; 147,84; 147,63 and 144,44 (C=N); 146,51 ; 145,34; 141 ,39; 131 ,92; 130,71 (d, J: 9,41 Hz); 130,15 (d, J: 3,14 Hz); 128,71 ; 127,40; 1 17,46 (d, J: 23,15 Hz); 1 12,13; 1 1 1 ,76; 35,67 and 35,62; 28,55; 15,82

- ESI-HRMS: mass: m/z 449.1321 , calculated [M+H] ⁺ : m/z 450.1400, found [M+H] ⁺ : m/z 450.1406, calculated [M- H] ⁺ : m/z 448.1243, found [M-H] ⁺ : m/z 448.1259; calculated for C ₂₃ H ₂₀ FN ₅ O ₂ S. 0.5 C ₂ H ₅ -OH 61%C 4.91%H 14.82%N, Found: 60.71 %C 4.877%H 15.09%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(2,4- dinitrophenyl)methylidine]acetohydrazide (Formula 2f)

When R" = in Formula 2a-t, the molecule shown in Formula 2f is obtained.

It was synthesized as a yellow powder substance with a yield of 60-95% and its melting temperature is in the range of 121 - 123.5°C.

FTIR v (cm ¹ ): 3470 (N-H stretching band), 3089 (ar C-H stretching band), 2933, 2833 (aldehyde C-H asymmetric and symmetric stretching band), 1681 (amide C=O stretching band), 1599 (hydrazone C=N stretching band), 1544 (N-0 asymmetric stretching band of the nitro structure), 1509 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1438 (aldehyde C-H asymmetric and symmetric stretching band), 1339 (N-0 symmetric stretching band of nitro structure), 1225 (C-S stretching band), 1 153 (aromatic C-F stretching band), 838 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 12.36 and 12.18 (2s, 1 H, CONH); 8.78 (s, 1 H, aromatic C ₃ -H); 8.55 (d, 1 H, J: 8.65 Hz, aromatic C ₅ -H); 8.32 (d, 1 H, J: 8.65 Hz, aromatic C ₅ -H); 8.67 and 8.47 (2s, 1 H, N=CH); 7.77 (s, 1 H, furan C ₅ -H); 7.63-7.60 (m, 2H, FPhC ₂ , ₆ -H); 7.48 (t, J: 8,72 Hz, 2H, FPhC ₃ , ₅ -H); 6.53 (s, 1 H, furan C ₄ -H); 6.20 (s, 1 H, furan C ₃ -H); 4,48 and 4,12 (2s, 2H, SCH ₂ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 169.54 and 164.54 (C=O), 163.22 (d, J: 248.28 Hz), 151.60, 148.02, 147.87, 147.65, 145.35, 141.60 and 138.38 (C=N), 141.32, 134.32, 130.68 (d, J: 9.19 Hz), 130.09 (d, J: 2.98 Hz), 129.98, 127.99, 120.78, 1 17.49 (d, J: 23.51 Hz), 1 12.1 1 , 1 1 1.80, 35.59 and 35.30 ESI-HRMS: mass: m/z 511.0710, calculated [M+H] ⁺ : m/z 512.0788, found [M+H] ⁺ : m/z 512.0797, calculated [M- H] ⁺ : m/z 510.0632, found [M-H] ⁺ : m/z 510.0648; calculated for C21H14FN7O6S.O.5 C2H5-OH 49.44%C 3.21 %H 18.34%N, Found: :49.45%C 3.224%H 18.27%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1,2,4-triazol-3- yl]sulfanyl]-N'-[(3,4,5- trimethoxyphenyl)methylidene]acetohydrazide (Formula 2g)

When R" = in Formula 2a-t, the molecule shown in Formula 2g is obtained.

Formula 2g

It was synthesized as a white powder substance with a yield of 60-95% and its melting temperature is between 176-179°C.

- FTIR v (cm- ¹ ): 3148 (N-H stretching band), 3072 (ar C-H stretching band), 2978, 2935, 2872 (aldehyde C-H asymmetric and symmetric stretching band), 1650 (amide C=O stretching band), 1600 (hydrazone C=N stretching band), 1509 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1436, 1372 (aldehyde C-H asymmetric and symmetric stretching band), 1223 (C-S stretching band), 1 125 (aromatic C-F stretching band), 846 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1.74 and 1 1.69 (2s, 1 H, CONH); 8.12 and 7.92 (2s, 1 H, N=CH); 7.78 (d, J: 1.8 Hz, 1 H, furan C5-H); 7.64-7.60 (m, 2H, FPhC ₂ ,6-H); 7.50-7.45 (m, 2H, FPhC ₃ ,5-H); 7.01 and 6.99 (2s, 2H, aromatic C ₂ , ₆ -H); 6.54 (dd, J: 3.5, 1 .8 Hz, 1 H, furan C ₄ -H); 6.24 (d, J: 3.5 Hz, 1 H, furan C3-H); 4.46 and 4.08 (2s, 2H, SCH ₂ ); 3.81 ve 3.83 (2s, 6H, Ph ₃ ,5-OCH ₃ ); 3.69 and 3.71 (2s, 3H, Ph4-OCH ₃ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168.93 and 163.66 (C=O), 163.20 (d, J: 248.1 1 Hz), 153.61 , 151.72, 147.85, 147.63 and 144.13 (C=N), 145.31 , 141.36, 139.59, 130.68 (d, J: 9.18 Hz), 130.13 (d, J: 3.05 Hz), 129.92, 1 17.43 (d, J: 23.5 Hz), 1 12.12, 11 1.77, 104,65, 60.57, 56.36, 35.63 and 35.16

- ESI-HRMS: mass: m/z 511.1325, calculated [M+H] ⁺ : m/z 512.1403, found [M+H] ⁺ : m/z 512.1415, calculated [M- H] ⁺ : m/z 510.1247, found [M-H] ⁺ : m/z 510.1263; calculated for C24H22FN5O5S.O.5H2O: 55.38%C 4.45%H 13.45%N, Found: 55.21 %C 4.683%H 13.25%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1,2,4-triazol-3- yl]sulfanyl]-N'-[(4-nitrophenyl)methylidene]acetohydrazide (Formula 2h)

When R" = in Formula 2a-t, the molecule shown in Formula 2h is obtained.

Formula 2h

It was synthesized as a light-yellow powder substance with a yield of 60-95% and its melting temperature is between 107- 109°C.

FTIR v (cm ¹ ): 3155 (N-H stretching band), 3026 (ar C-H stretching band), 2971 , 2877 (aldehyde C-H asymmetric and symmetric stretching band), 1687 (amide C=O stretching band), 1589 (hydrazone C=N stretching band), 1567 (N-0 asymmetric stretching band of the nitro structure), 1509 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1453, 1340 (aldehyde C-H asymmetric and symmetric stretching band), 1340 (N-0 symmetric stretching band of the nitro structure), 1233 (C-S stretching band), 1 185 (aromatic C-F stretching band), 839 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1 .97 (s, 1 H, CONH); 8.27 (d, J: 8,9 Hz, 2H, aromatic C ₃ , ₅ -H); 8.1 1 (s, 1 H, N=CH); 7.93 (d, J: 8,9, 2H, aromatic C _2,6 -H); 7.76 (d, J: 1.8 Hz, 1 H, furan C ₅ -H); 7.64-7.60 (m, 2H, FPhC ₂ ,6- H); 7.47 (t, J: 8,7 Hz, 2H, FPhC ₃ ,5-H); 6.53 (dd, J: 3.5, 1.8 Hz, 1 H, furan C ₄ -H); 6.22 (d, J: 3.5 Hz, 1 H, furan C ₃ - H); 4.5 and 4.1 1 (2s, 2H, SCH ₂ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 169.32 and 164.26 (C=O), 163.21 (d, J: 248.18 Hz), 151.76, 148.18, 147.88, 145.32, 145.19 and 142.05 (C=N), 141.31 , 140.62, 130.66 (d, J: 9.15 Hz), 130.09 (d, J: 2.76 Hz), 128.23, 124.45, 1 17.48 (d, J: 23.56 Hz), 1 12.1 1 , 1 1 1.82, 35.62 and 35.45

- ESI-HRMS: mass: m/z 466.0859, calculated [M+H] ⁺ : m/z 467.0937, found [M+H] ⁺ : m/z 467.0943, calculated [M- H] ⁺ : m/z 465.0781 , found [M-H] ⁺ : m/z 465.0796; calculated for C21H15FN6O4S.2H20: 50.20%C 3.81 %H 16.73%N, Found: 49.94%C 3.829%H 16.19%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- methylphenyl)methylidene]acetohydrazide (Formula 2i)

When R" = in Formula 2a-t, the molecule shown in Formula 2i is obtained.

Formula 2i

It was obtained as a white powder with a yield of 60-95% and its melting temperature is in the range of 167-169.5°C.

FTIR v (cm ¹ ): 3167 (N-H stretching band), 3080 (ar C-H stretching band), 2959, 2924 (aldehyde C-H asymmetric and symmetric stretching band), 1671 (amide C=O stretching band), 1617 (hydrazone C=N stretching band), 1508 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1429, 1382 (aldehyde C-H asymmetric and symmetric stretching band), 1222 (C-S stretching band), 1 155 (aromatic C-F stretching band), 843 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1.69 and 1 1.61 (s, 1 H, CONH); 8.15 and 7.97 (s, 1 H, N=CH); 7.77 (d, J: 1 .8 Hz, 1 H, furan C ₅ -H); 7.63-7.59 (m, 2H, FPhC ₂ ,6-H); 7.55 (d, J: 8.07 Hz, 2H, aromatic C 2,6 -H); 7.47 (td, J: 8.7; 1 .72 Hz, 2H, FPhC ₃ ,5-H); 7.24 (d, J: 8.07 Hz, 2H, aromatic C 3,5 -H); 6.53 (dd, J: 3.57, 1 .71 Hz, 1 H, furan C ₄ - H); 6.22 (d, J: 3.57 Hz, 1 H, furan C ₃ -H); 4.46 and 4,06 (2s, 2H, SCH ₂ ), 2.33 (s, 3H, PhCH ₃ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168.77 and 163.58 (C=O), 163.20 (d, J: 248.23 Hz), 151.96, 147.84, 147.67 and 144.48 (C=N), 145.32, 141.40, 140.28, 131.67, 130.70 (d, J: 9.19 Hz), 130.15 (d, J: 2.75 Hz), 129.89, 127.31 , 1 17.45 (d, J: 23.46 Hz), 1 12.12, 1 1 1.77, 35.69 and 35.63, 21.49.

- ESI-HRMS: mass: m/z 435.1 165, calculated [M+H] ⁺ : m/z 436.1243, found [M+H] ⁺ : m/z 436.1249, calculated [M- H] ⁺ : m/z 434.1087, found [M-H] ⁺ : m/z 434.1102; calculated for C22H18FN5O2S.H2O: 58.27%C 4.45%H 15.44%N, Found: 57.98%C 4.228%H 15.09%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(3,4-dimethoxy phenyl)methylidene]acetohydrazide (Formula 2j)

When R" = in Formula 2a-t, the molecule shown in Formula 2j is obtained.

Formula 2j It was synthesized as a white powder substance with a yield of 60-95% and its melting temperature is between 200- 202.5°C.

- FTIR v (cnr ¹ ): 3168, 3108 (N-H stretching band), 3082, 3029, 3003 (ar C-H stretching band), 2981 , 2925, 2837 (aldehyde C-H asymmetric and symmetric stretching band), 1678 (amide C=O stretching band), 1598 (hydrazone C=N stretching band), 1509 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1462, 1435, 1385 (aldehyde C-H asymmetric and symmetric stretching band), 1221 (C-S stretching band), 1 143 (aromatic C-F stretching band), 844 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1 .62 and 1 1 .56 (2s, 1 H, CONH); 8.1 1 and 7.92 (s, 1 H, N=CH); 7.77 (d, J: 1.8 Hz, 1 H, furan C ₅ -H); 7.63-7.59 (m, 2H, FPhC ₂ ,s-H); 7.49-7.45 (m, 2H, FPhC ₃ ,5-H); 7.30 (d, 1 H, J: 1.94 Hz, aromatic C ₂ -H); 7.22 (dd, 1 H, J: 8.31 ; 1 .94 Hz, aromatic C ₆ -H); 7.02 (d, 1 H, J: 8.31 Hz, aromatic C ₅ -H); 6.53 (dd, J: 3.5, 1 .8 Hz, 1 H, furan C ₄ -H); 6.22 (d, J: 3.5 Hz, 1 H, furan C ₃ -H); 4.45 and 4.05 (2s, 2H, SCH ₂ ), 3.8 (s, 3H, Ph-3OCH ₃ ); 3.79 (s, 3H, Ph-4OCH ₃ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168.70 and 163.40 (C=O), 163.20 (d, J: 248.12 Hz), 151.72, 151.05, 149.43, 147.84, 147.79 and 144.36 (C=N), 145.32, 141.39, 130.71 (d, J: 9.18 Hz), 130.15 (d, J: 3.24 Hz), 127.1 1 , 121.56, 1 17.45 (d, J: 22.99 Hz), 1 12.13, 1 1 1.98, 1 1 1.84, 109.21 , 56,01 , 55.89, 35.67 and 35.35

- ESI-HRMS: mass: m/z 481.1220, calculated [M+H] ⁺ : m/z 482.1298, found [M+H] ⁺ : m/z 482.1304, calculated [M- H] ⁺ : m/z 480.1 141 , found [M-H] ⁺ : m/z 480.1159; calculated forC ₂₃ H ₂ oFN ₅ 04S: 57.37%C 4.19%H 14.54%N, Found: 57.21 %C 4.370%H 14.53%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4-methoxy)methylidene]aceto hydrazide

(Formula 2k)

When RII = in Formula 2a-t, the molecule shown in Formula 2k is obtained.

Formula 2k

It was synthesized as a white powder substance with a yield of 60-95% and its melting temperature is between 194- 195.5°C. FTIR v (cnr ¹ ): 3105 (N-H stretching band), 3078 (ar C-H stretching band), 2966 (aldehyde C-H asymmetric and symmetric stretching band), 1681 (amide C=O stretching band), 1666 (hydrazone C=N stretching band), 1602 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1453, 1377 (aldehyde C-H asymmetric and symmetric stretching band), 1221 (C-S stretching band), 1 173 (aromatic C-F stretching band), 831 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1 .54 and 1 1 .47 (s, 1 H, CONH); 8.05 and 7.87 (s, 1 H, N=CH); 7.70 (d, J: 1 .7 Hz, 1 H, furan C ₅ -H); 7.58-7.64 (m, 2H, FPhC ₂ , ₆ -H); 7.53 (d, J: 8,76 Hz, 2H, aromatic C ₂ , ₆ -H); 7.39 (t, J: 8,7 Hz, 2H, FPhC ₃ ,5-H); 6.92 (d, J: 8,76 Hz, 2H, aromatic C ₃ ,5-H); 6.46 (dd, J: 3.5, 1 .7 Hz, 1 H, furan C ₄ -H); 6.14 (d, J: 3.5 Hz, 1 H, furan C ₃ -H); 4.37 and 3,97 (2s, 2H, SCH ₂ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168.64 and 163.44 (C=O), 163.21 (d, J: 248.32 Hz), 161.21 , 151.74, 147.84, 147.53 and 144.28 (C=N), 145.31 , 141.40, 130.69 (d, J: 9.18 Hz), 130.15 (d, J: 2.74 Hz), 128.92, 126.95, 1 17.45 (d, J: 23.39 Hz), 1 14.76, 1 12.12, 11 1.77, 55.73, 35.70 and 35.66

- ESI-HRMS: mass: m/z 451.1 114, calculated [M+H] ⁺ : m/z 452.1 192, found [M+H] ⁺ : m/z 452.1 174, calculated [M- H] ⁺ : m/z 450.1036, found [M-H] ⁺ : m/z 450.1040; calculated for C ₂₂ Hi8FN ₅ O ₃ S: 58.53%C 4.02%H 15.51%N, Found: 58.41 %C 3.953%H 15.32%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- cyanophenyl)methylidene]acetohydrazide (Formula 2I)

When R" = in Formula 2a-t, the molecule shown in Formula 21 is obtained.

Formula 2I

It was synthesized as a white powder substance with a yield of 60-95% and its melting temperature is between 232- 234.5°C.

FTIR v (cm ¹ ): 3190 (N-H stretching band), 3101 , 301 1 (ar C-H stretching band), 2929, 2864 (aldehyde C-H asymmetric and symmetric stretching band), 2221 (C=N stretching band), 1681 (amide C=O g.b), 1575 (hydrazone C=N stretching band), 1508 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1454, 1361 (aldehyde C-H asymmetric and symmetric stretching band), 1227 (C-S stretching band), 1 153 (aromatic C-F stretching band), 839 (1 ,4-disubstituted aromatic C-H stretching band);

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 12.02 and 1 1.92 (2s, 1 H, CONH); 8.26 and 8.05 (2s, 1 H, N=CH); 7.88 (d, J=8.1 Hz, 2H, CN-PhC2, ₆ -H); 7.85 (d, J: 8.1 Hz, 2H, CN-PhC ₃ ,5-H); 7.77 (d, J: 1.8 Hz, 1 H, furan C ₅ -H); 7.64- 7.60 (m, 2H, FPhC ₂ ,6-H); 7.47 (t, J: 8.7 Hz, 2H, FPhC ₃ ,5-H); 6.53 (dd, J: 3.5, 1.8 Hz, 1 H, furan C ₄ -H); 6.24 (d, J: 3.5 Hz, 1 H, furan C ₃ -H); 4.50 and 4.12 (2s, 2H, SCH ₂ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 6 (ppm): 169.27 (C=O), 163.18 (d, J: 242.35 Hz), 151.78, 147.87, 145.64 ve 142.42 (C=N), 145.33, 141.36, 138.81 , 133.16, 130.68 (d, J: 9.23 Hz), 130.12 (d, J: 2.71 Hz), 127.89, 1 19.1 1 , 1 17.47 (d, J: 23.02 Hz), 1 12.26, 1 12.12, 1 1 1.79, 35.63 and 35.51

- ESI-HRMS: mass: m/z 446.0961 , calculated [M+H] ⁺ : m/z 447.1039, found [M+H] ⁺ : m/z 447.1043, calculated [M- H] ⁺ : m/z 445.0883, found [M-H] ⁺ : m/z 445.0898; calculated for C ₂₂ HI ₅ FN ₆ O ₂ S. 0.5H ₂ O: 58.02%C 3.54%H 18.45%N, Found: 58.49%C 3.506%H 18.47%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- hydroxyphenyl)methylidene]acetohydrazide (Formula 2m)

When R" = in Formula 2a-t, the molecule shown in Formula 2m is obtained.

It was synthesized as a white powder substance with a yield of 60-95% and its melting temperature is between 264.5- 267°C.

FTIR v (cm ¹ ): 3202 (N-H stretching band), 3064 (ar C-H stretching band), 2883, 2804 (aldehyde C-H asymmetric and symmetric stretching band), 1659 (amide C=O stretching band), 1603 (hydrazone C=N stretching band), 1509 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1451 , 1377 (aldehyde C-H asymmetric and symmetric stretching band), 1224 (C-S stretching band), 1 144 (aromatic C-F stretching band), 849 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1 .55 and 1 1 .48 (2s, 1 H, CONH); 9.93 (s, 1 H, OH); 8.08 and 7.91 (2s, 1 H, N=CH); 7.77 (d, J: 1 .7 Hz, 1 H, furan C ₅ -H); 7.64-7.60 (m, 2H, FPhC ₂ , ₆ -H); 7.53 (d, J: 8.62 Hz, 2H, OH-PhC ₂ , ₆ - H); 7.48-7.45 (m, 2H, FPhC ₃ , ₅ -H); 6.82 (d, 2H, J: 8.62 Hz, OH-PhC ₃ , ₅ -H); 6.53 (dd, J: 3.5; 1.8 Hz, 1 H, furan C ₄ - H); 6.24 (d, J: 3.5 Hz, 1 H, furan C ₃ -H); 4.45 and 4.05 (2s, 2H, SCH ₂ ) ¹³ C-NMR (APT) (125 MHz) (DMSO-ds) 5 (ppm): 168.49 and 163.28 (C=O), 163.2 (d, J: 248.57 Hz), 159.80, 151.73, 147.82, 147.94 and 144,71 (C=N), 145.32, 141.38, 130.71 (d, J: 9.18 Hz), 130.14 (d, J: 3.15 Hz), 129.07, 125.38, 117.45 (d, J: 23.09 Hz), 116.17, 112.13, 111.78, 35.70

ESFHRMS: mass: m/z 437.0957, calculated [M+Hp: m/z 438.1036, found [M+H] ⁺ : m/z 438.1038, calculated [M- Hp: m/z 436.0879, found [M-Hp: m/z 436.0895; calculated for C ₂₁ H ₁₆ FN ₅ O ₃ S. 0.5 HzO: 56.5%C 3.84%H 15.69%N, Found: 56.53%C 3.748%H 15.62%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1,2,4-triazol-3- y1]sulfanyl]-N ^, -[(3- hydroxyphenyl)methylldene]acetohydrazlde (Formula 2n)

When R In Formula 2a-t, the molecule shown In Formula 2n Is obtained.

It was synthesized as a white powder substance with a yield of 60-95% and Its melting temperature Is between 232- 234.5°C.

FTIR v (cm ^-1 ): 3157 (N-H stretching band), 3073 (ar C-H stretching band), 2962, 2830 (aldehyde C-H asymmetric and symmetric stretching band), 1666 (amide C=O stretching band), 1612 (hydrazone C=N stretching band), 1510 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1440, 1383 (aldehyde C-H asymmetric and symmetric stretching band), 1225 (C-S stretching band), 1155 (aromatic C-F stretching band), 841 (1 ,4-dlsubstituted aromatic C-H stretching band)

¹ H-NMR (500 MHz) (DMSO-cfa) 5 (ppm): 11.64 (s, 1 H, CONH); 9.65 (2s, 1 H, OH); 8.09 ve 7.93 (2s, 1 H, N=CH); 7.77 (d, J: 1.65 Hz, 1 H, furan C ₅ -H); 7.63-7.61 (m, 2H, FPhCzj-H); 7.47 (t, J: 8.7 Hz, 2H, FPhCs, ₅ -H); 7.24 (t, J: 7.81 Hz, aromatic C ₅ -H); 7.16 (s, 1 H, aromatic Cz-H); 7.08 (d, J: 7.81 Hz, 1 H, aromatic C ₅ -H); 6.82 (d, J: 7.81 Hz, 1 H, aromatic C<-H); 6.53 (dd, J: 3.53; 1.65 Hz, 1 H, furan C<-H); 6.24 (d, J: 3.53 Hz, 1 H, furan C ₅ -H); 4.49 and 4.06 (2s, 2H, SCHz)

^1S C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168.79 and 163.69 (C=O), 163.22 (d, J: 248.72 Hz), 158.13, 151.73, 147.83, 147.73 and 144.69 (C=N), 145.30, 141.38, 135.60, 130.69 (d, J: 9.19 Hz), 130.35, 130.11 (d, J: 3.21 Hz), 119.00, 117.88, 117.47 (d, J: 23.40 Hz), 113.02, 112.12, 111.80, 35.71

ESFHRMS: mass: m/z 437.0957, calculated [M+Hp: m/z 438.1036, found [M+Hp: m/z 438.1039, calculated [M- Hp: m/z 436.0879, found [M-Hp: m/z 436.0895; calculated for CziHisFNsOsS: 57.66%C 3.69%H 16.01%N; Found: 57.19%C 3.62%H 15.81%N. 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1,2,4-triazol-3-yl] sulfanyl]-N'-[(4- chlorophenyl)methylidene]acetohydrazide (Formula 2o)

When R" = in Formula 2a-t, the molecule shown in Formula 2o is obtained.

Formula 2o

It was synthesized as a white powder substance with a yield of 60-95% and its melting temperature is between 1 17-119°C.

FTIR v (cm ¹ ): 3178 (N-H stretching band), 3080 (ar C-H stretching band), 2954 (aldehyde C-H asymmetric and symmetric stretching band), 1672 (amide C=O stretching band), 1615 (hydrazone C=N stretching band), 1509 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1448, 1382 (aldehyde C-H asymmetric and symmetric stretching band), 1223 (C-S stretching band), 1 125 (aromatic C-F stretching band), 843 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1.82 and 1 1.74 (2s, 1 H, CONH); 8.20 and 8.01 (2s, 1 H, N=CH); 7.77 (m, 1 H, furan C ₅ -H); 7.73 (d, J: 8.59 Hz, 2H, CIPhC2, ₆ -H), 7.63-7.60 (m, 2H, FPhC ₂ ,6-H), 7.52-7.45 (m, 4H, FPhC ₃ ,5-H ve CIPhC ₃ ,5-H); 6.53 (m, 1 H, furan C ₄ -H); 6.24 (d, J: 3.58 Hz, 1 H, furan C ₃ -H); 4.47 and 4.08 (2s, 2H, SCH ₂ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d6) 5 (ppm): 168.97 and 163.85 (C=O), 163.21 (d, J: 248.18 Hz), 151.89, 147.85, 146.34 and 143.13 (C=N), 145.32, 141.36, 134.92, 133.30, 130.68 (d, J: 9.22 Hz), 130.12 (d, J: 3.22 Hz), 129.35, 128.95, 1 17.46 (d, J: 23.30 Hz), 1 12.12, 1 1 1.80, 35.65 and 35.58

- ESI-HRMS: mass: m/z 455.0619, calculated [M+H] ⁺ : m/z 456.0697, found [M+H] ⁺ : m/z 456.0703, calculated [M- H] ⁺ : m/z 454.0540, found [M-H] ⁺ : m/z 454.0557; calculated for C21H15CIFN5O2S.I H2O: 53.22%C 3.62%H 14.78%N, Found: 52.9%C 3.631 %H 14.53%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1,2,4-triazol-3- yl]sulfanyl]-N'-[(4- fluorophenyl)methylidene]acetohydrazide (Formula 2p)

When R" = in Formula 2a-t, the molecule shown in Formula 2p is obtained.

Formula 2p

It was synthesized as a white powder substance with a yield of 60-95% and its melting temperature is between 129-131 °C.

FTIR v (cm ¹ ): 3180 (N-H stretching band); 3078 (ar C-H stretching band); 2964 (aldehyde C-H asymmetric and symmetric stretching band), 1671 (amide C=O stretching band); 1600 (hydrazone C=N stretching band), 1509 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands); 1427, 1383 (aldehyde C-H asymmetric and symmetric stretching band), 1232 (C-S stretching band), 1 155 (aromatic C-F stretching band), 839 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 6 (ppm): 1 1.7 (2s, 1 H, CONH); 8.19 and 8.01 (2s, 1 H, N=CH); 7.75-7.72 (m, 2H, N=C-FPhC2, ₆ -H); 7.63-7.60 (m, 2H, FPhC ₂ ,6-H); 7.78 (m, 1 H, furan C ₅ -H); 7.47 (t, J: 8.74 Hz, 2H, FPhC ₃ ,5-H); 7.31 -7.26 (m, 2H, N=C-FPhC ₃ ,5-H); 6.53 (dd, J: 3.57; 1.77 Hz, 1 H, furan C ₄ -H); 6.23 (d, J: 3.57 Hz, 1 H, furan C ₃ - H); 4.47 and 4.07 (2s, 2H, SCH2)

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168.90 and 163.75 (C=O), 163.50 (d, J: 247.76 Hz), 163.20 (d, J: 248.61 Hz), 151.70, 147.85, 146.52 and 143.25 (C=N), 145.32, 141.37, 130.99 (d, J: 2.78 Hz), 130.69 (d, J: 9.23 Hz), 130.13 (d, J: 3.23 Hz), 129.51 (d, J: 8.70 Hz), 1 17.46 (d, J: 23.54 Hz), 1 16.33 (d, J: 22.05 Hz), 1 12.12, 1 1 1.79, 35.65 and 35.59

- ESI-HRMS: mass: m/z 439.0914, calculated [M+H] ⁺ : m/z 440.0992, found [M+H] ⁺ : m/z 440.0976, calculated [M- H] ⁺ : m/z 438.0836, found [M-H] ⁺ : m/z 438.0840; calculated for C21H15F2N5O2S. H2O: 55.14%C 3.75%H 15.31 %N, Found: 55.22%C 4.025%H 15.21%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4- bromophenyl)methylidene]acetohydrazide (Formula 2r) in Formula 2a-t, the molecule shown in Formula 2r is obtained.

Formula 2r It was synthesized as a white powder substance with a yield of 60-95% and its melting temperature is between 120-122°C.

FTIR v (cm ¹ ): 3178 (N-H stretching band), 3079 (ar C-H stretching band), 2961 , 2925 (aldehyde C-H asymmetric and symmetric stretching band), 1671 (amide C=O stretching band), 1615 (hydrazone C=N stretching band), 1509 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1447, 1381 (aldehyde C-H asymmetric and symmetric stretching band), 1222 (C-S stretching band), 1 125 (aromatic C-F stretching band), 842 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1.88 and 1 1.79 (2s, 1 H, CONH); 8.23 and 8.04 (2s, 1 H, N=CH); 7.83 (d, J: 1 .67 Hz, 1 H, furan C ₅ -H); 7.71 -7.65 (m, 6H, FPhC ₂ , ₆ -H ve BrPhC2, ₃ ,5,6-H ); 7.52 (t, J: 8.69 Hz, 2H, FPhC ₃ , ₅ - H); 6.58 (dd, J: 3.58; 1 .67 Hz, 1 H, furan C ₄ -H); 6.29 (d, J: 3.58 Hz, 1 H, furan C ₃ -H); 4.52 and 4.13 (2s, 2H, SCH ₂ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168.97 and 163.86 (C=O), 163.20 (d, J: 248.78 Hz), 151.88, 147.85, 146.43 and 143.23 (C=N), 145.32, 141.35, 133.64, 132.27, 130.68 (d, J: 9.18 Hz), 130.1 1 (d, J: 2.76 Hz), 129.18, 123.70, 1 17.47 (d, J: 23.02 Hz), 1 12.12, 1 11.80, 35.66 and 35.58

- ESI-HRMS: mass: m/z 501.0239, calculated [M+H] ⁺ : m/z 502.0133, found [M+H] ⁺ : m/z 502.0178, calculated [M-H] ⁺ : m/z 500.0161 , found [M-H] ⁺ : m/z 500.0033; calculated for C2iHi ₅ BrFN ₅ O2S. 0,5H ₂ O: 49.52%C 3.17%H 13.75%N, Found: 49.08%C 3.468%H 13.12%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(2-hydroxyphenyl)methylidene ]acetohydrazide

(Formula 2s)

When R" = in Formula 2a-t, the molecule shown in Formula 2s is obtained.

It was synthesized as a white powder substance with a yield of 60-95% and its melting temperature is between 191 - 192.5°C.

- FTIR v (cm- ¹ ): 3161 (N-H stretching band), 3010 (ar C-H stretching band), 2969, 2928, 2853 (aldehyde C-H asymmetric and symmetric stretching band), 1672 (amide C=O stretching band), 1617 (hydrazone C=N stretching band), 151 1 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1453, 1386 (aldehyde C-H asymmetric and symmetric stretching band), 1225 (C-S stretching band), 1 154 (aromatic C-F stretching band), 844 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1.53 and 10.88 (2s, 1 H, CONH); 10.01 (s, 1 H, OH); 8.34 ve 8.23 (2s, 1 H, N=CH); 7.69 (d, J: 1.7 Hz, 1 H, furan C ₅ -H); 7.57 (dd, 1 H, J: 7.67; 1.5 Hz, OHPhCe-H); 7.55-7.52 (m, 2H, FPhC ₂ ,6-H); 7.39 (td, J: 8,7; 1 ,95 Hz, 2H, FPhC ₃ , ₅ -H); 7.22 (td, 1 H, J: 7,67; 1 ,5 Hz, OHPhC ₄ -H); 6.84 (d, 1 H, J: 7,67 Hz, OHPhC ₃ -H); 6.78 (t, 1 H, J: 7,67 Hz, OHPhC ₅ -H); 6.46 (dd, J: 3.6; 1.8 Hz, 1 H, furan C ₄ -H); 6.17 (d, J: 3.7 Hz, 1 H, furan C ₃ -H); 4.38 ve 3.99 (2s, 2H, SCH ₂ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168.55 and 163.62 (C=O), 163.25 (d, J: 248.75 Hz), 157.78, 151.65, 147.96, 147.83 and 141.95 (C=N), 145.36, 141.31 , 131.98, 130.69 (d, J: 9.16 Hz), 130.13 (d, J: 2.79 Hz), 129.67, 1 19.83, 1 19.05, 1 17.49 (d, J: 23.01 Hz), 1 16.83, 1 12.14, 11 1.89, 35.71 and 35.37

- ESI-HRMS: mass: m/z 437.0957, calculated [M+H] ⁺ : m/z 438.1036, found [M+H] ⁺ : m/z 438.1018, calculated [M- H] ⁺ : m/z 436.0879, found [M-H] ⁺ : m/z 436.0883; calculated for C21H16FN5O3S: 57.66%C 3.69%H 16.01%N, Found: 57.12%C 3.932%H 15.81%N.

2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(4-dimethyl aminophenyl)methylidene]acetohydrazide (Formula 2t)

When in Formula 2a-t, the molecule shown in Formula 2t is obtained.

Formula 2t

It was synthesized as a white powder substance with a yield of 60-95% and its melting temperature is between 257- 258.5°C.

FTIR v (cm ¹ ): 3161 (N-H stretching band), 3074 (ar C-H stretching band), 2895 (aldehyde C-H asymmetric and symmetric stretching band), 1666 (amide C=O stretching band), 1607 (hydrazone C=N stretching band), 1510 (aromatic C=C stretching, amide II N-H bending and C-N stretching combination bands), 1445, 1384 (aldehyde C-H asymmetric and symmetric stretching band), 1221 (C-S stretching band), 1 151 (aromatic C-F stretching band), 844 (1 ,4-disubstituted aromatic C-H stretching band)

- ¹ H-NMR (500 MHz) (DMSO-d ₆ ) 5 (ppm): 1 1.44 and 1 1.39 (2s, 1 H, CONH); 8.03 and 7.87 (2s, 1 H, N=CH); 7.78 (d, J: 1.7 Hz, 1 H, furan C ₅ -H); 7.63-7.59 (m, 2H, FPhC ₂ ,6-H); 7.51 -7.44 (m, 6H, FPhC ₃ ,5-H and N(CH ₃ ) ₂ PhC ₂ , 3,5,6- H); 6.54 (dd, J: 3.5; 1.7 Hz, 1 H, furan C ₄ -H); 6.24 (d, J: 3.5 Hz, 1 H, furan C ₃ -H); 4.43 and 4.03 (2s, 2H, SCH ₂ ), 2.97 (s, 6H, -N(CH ₃ ) ₂ )

- ¹³ C-NMR (APT) (125 MHz) (DMSO-d ₆ ) 5 (ppm): 168.26 and 162.97 (C=O), 163.20 (d, J: 248.36 Hz), 152.03, 151.88, 148.41 and 145.18 (C=N), 147.82, 145.32, 141.43, 130.71 (d, J: 9.23 Hz), 130.18 (d, J: 2.78 Hz), 128.62, 121.68, 1 17.45 (d, J: 22.96 Hz), 1 12.22, 1 12.13, 1 11.83, 40.20, 35.76 and 35.73

- ESI-HRMS: mass: m/z 464.1430, calculated [M+H] ₊ : m/z 465.1509, found [M+H] ⁺ : m/z 465.1515, calculated [M- H] ⁺ : m/z 463.1352, found [M-H] ⁺ : m/z 463.1369; calculated for C ₂₃ H ₂ I FN ₆ O ₂ S: 59.47%C 4.56%H 18.09%N, Found: 59.40%C4.771 %H 18.04%N. The synthesis phases of 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide (Formula 1 ) and 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(substituted phenyl)methylidene]acetohydrazide (Formula 2a) that are the subject of the invention and their derivatives (Formula 2a-t) are shown below as Reaction i, Reaction ii, Reaction iii, Reaction iv, Reaction v, respectively.

Formula 1 Formula 2a-t

The synthesis method of 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide (Formula 1 ), which is one of the molecules that are the subject of the invention, comprises the process steps of

I. obtaining N-(4-fluorophenyl)-2-(furan-2-carbonyl)hydrazine-1 -carbotioamide as a result of adding 0.03 mol 4- fluorophenyl isothiocyanate to the absolute ethanol solution of 0.03 mol furan-2-carbohydrazide and heating it under reflux in a water bath for 1 -4 hours, filtering and drying the precipitate formed after 1 day (Reaction i), ii. obtaining 4-(4-Fluorophenyl)-5-(furan-2-yl)-2,4-dihydro-3H-1 ,2,4-triazole-3-thion as a result of heating N-(4- fluorophenyl)-2-(furan-2-carbonyl)hydrazine-1 -carbotioamide in 2N sodium hydroxide (NaOH) under reflux for 2- 4 hours and precipitation of the resulting 4-(4-fluorophenyl)-5-(furan-2-yl)-2,4-dihydro-3H-1 ,2,4-triazole-3-thion with 12.5% hydrochloric acid (HCI) and then washing the precipitated product with distilled water and drying it (Reaction ii), iii. obtaining ethyl ((4-fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl)sulfanyl)acetate as a result of preparating a suspension of 0.005 mol 4-(4-fluorophenyl)-5-(furan-2-yl)-2,4-dihydro-3H-1 ,2,4-triazole-3-thion in ethanol and adding 0.005 mol potassium carbonate (K2CO3) and 0.0055 mol ethyl bromoacetate to the said suspension and heating in a water bath under reflux for 5-6 hours, then pouring the reaction medium into ice water and keeping it in the refrigerator for 1 day and then filtering and drying the precipitated ethyl ((4-fluorophenyl)-5-(furan-2-yl)-4H- 1 ,2, 4-triazol-3-yl)sulfanyl)acetate. (Reaction iii), and iv.obtaining 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide molecule (Formula 1 ) that is the subject of the invention as a result of heating 0.005 mol ethyl ((4-fluoropheny l)-5-(fu ran-2-y l)-4H- 1 ,2,4- triazol-3-yl)sulfanyl)acetate and 0.028 mol of hydrazine hydrate under reflux in an ethanolic environment for 5-6 hours (Reaction iv).

N-(4-fluorophenyl)-2-(furan-2-carbonyl)hydrazine-1 -carbotioamide, 4-(4-Fluorophenyl)-5-(furan-2-yl)-2,4-dihydro-3H- 1 ,2,4-triazol-3-thion, ethyl ((4-fluoropheny l)-5-(fu ran-2-yl)-4H-1 ,2,4-triazol-3-yl)sulfanyl)acetate and 2-[[4-(4-Fluorophenyl)- 5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide molecule (Formula 1 ) that is the subject of the invention are obtained with an efficiency of 60-95%. The synthesis method of 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(substituted phenyl)methylidene]acetohydrazide derivatives (Formula 2a- t), which are the molecules that are the subject of the invention, comprises the process steps of

I. obtaining N-(4-fluorophenyl)-2-(furan-2-carbonyl)hydrazine-1 -carbotioamide as a result of adding 0.03 mol 4- fluorophenyl isothiocyanate to the absolute ethanol solution of 0.03 mol furan-2-carbohydrazide and heating it under reflux in a water bath for 1 -4 hours, filtering and drying the precipitate formed after 1 day (Reaction i), ii. obtaining 4-(4-Fluorophenyl)-5-(furan-2-yl)-2,4-dihydro-3H-1 ,2,4-triazole-3-thion as a result of heating N-(4- fluorophenyl)-2-(furan-2-carbonyl)hydrazine-1 -carbotioamide in 2N sodium hydroxide (NaOH) under reflux for 1 - 4 hours and precipitation of the resulting 4-(4-fluorophenyl)-5-(furan-2-yl)-2,4-dihydro-3H-1 ,2,4-triazole-3-thion with 12.5% hydrochloric acid (HCI) and then washing the precipitated product with distilled water and drying it (Reaction ii), iii.obtaining ethyl ((4-fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl)sulfanyl)acetate as a result of adding 0.005 mol potassium carbonate (K2CO3) and 0.0055 mol ethyl bromoacetate to a suspension of 0.005 mol 4-(4- fluorophenyl)-5-(furan-2-yl)-2,4-dihydro-3H-1 ,2,4-triazole-3-thion in ethanol and heating in a water bath under reflux for 5-6 hours, then pouring the reaction content into ice water and keeping it in the refrigerator for 1 day, and then, filtering and drying the precipitated ethyl ((4-fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3- yl)sulfanyl)acetate (Reaction iii), iv.obtaining 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide molecule (Formula 1 ) that is the subject of the invention as a result of heating 0.005 mol ethyl ((4-f luoropheny l)-5-(f u ran-2-y l)-4H-1 ,2,4- triazol-3-yl)sulfanyl)acetate and 0.028 mol of hydrazine hydrate under reflux in an ethanolic environment for 5-6 hours (Reaction iv), and v.obtaining the 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(substituted phenyl)methylidene]acetohydrazide derivatives (Formula 2a-t) that are subject of the invention in different reaction flasks as a result of adding 0.003 mol of aromatic aldehyde compound to the solution of 0.003 mol 2-[[4-(4- fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide (Formula 1 ) in absolute ethanol and boiling the reaction mixture under reflux in a water bath for 5-6 hours and further cooling, filtering the precipitated hydrazide-hydrazones and purifying them by washing with boiling ethanol.

N-(4-fluorophenyl)-2-(furan-2-carbonyl)hydrazine-1 -carbotioamide, 4-(4-Fluorophenyl)-5-(furan-2-yl)-2,4-dihydro-3H- 1 ,2,4-triazol-3-thion, ethyl ((4-f luoropheny l)-5-(fu ran-2-yl)-4H-1 ,2,4-triazol-3-yl)sulfanyl)acetate and 2-[[4-(4-Fluorophenyl)- 5-(fu ran-2-y l)-4H- 1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide molecule (Formula 1 ) are obtained with an efficiency of 60-95% (1.1 g).

Figure 1 shows the comparative column chart showing the effect of 24-hour application of 2-[[4-(4-Fluorophenyl)-5-(furan- 2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide (Formula 1 ) and and 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4- triazol-3-yl]sulfanyl]-N'-[(substituted phenyl)methylidene]acetohydrazide derivatives (Formula 2a-t), which are the molecules that are the subject of the invention, and cisplatin and osimertinib, which are used as standard, on the viability of MCF-7 cells. In other words, looking at Figure 1 , it is understood that the molecules that are the subject of the invention show inhibition levels close to standard drugs on MCF-7 breast cancer cell lines.

Figure 2 shows the comparative column chart showing the effect of 24-hour application of 2-[[4-(4-Fluorophenyl)-5-(furan- 2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide (Formula 1 ) and and 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4- triazol-3-yl]sulfanyl]-N'-[(substituted phenyl)methylidene]acetohydrazide derivatives (Formula 2a-t), which are the molecules that are the subject of the invention, and cisplatin and osimertinib, which are used as standard, on the viability of HEK293 cells. In Figure 2, the molecules that are the subject of the invention show toxic effects on HEK293 healthy cell lines at higher concentrations than standard drugs. In other words, the possible side effects of the molecules that are the subject of the invention are less than standard drugs.

Figures 3-101 show FTIR, ¹ H-NMR and ESI-HRMS (negative and positive) spectrums of 2-[[4-(4-Fluorophenyl)-5-(furan- 2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide (Formula 1 ) and 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3- yl]sulfanyl]-N'-[(substituted phenyl)methylidene]acetohydrazide) derivatives (Formula 2a-t), which are the molecules that are the subject of the invention.

Table 1. IC50 values of 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide (Formula 1 ) and 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'-[(substituted phenyl)methylidene]acetohydrazide) derivatives (Formula 2a-t), which are the molecules that are the subject of the invention and cisplatin and osimertinib used as standards on MCF-7 and HEK293 cells.

As can also be clearly seen from Table 1 , while the lethal effects of cisplatin and osimertinib molecules, which are highly effective molecules on the market, on normal cells are higher, the toxicity of the molecules subject to the invention is much less than the molecules in question, in other words, the possible side effects of the molecules subject to the invention are much lower than those of cisplatin and osimertinib molecules.

Table 2. Inhibition activities of 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide (Formula 1 ) and 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-y Ijsulfany l]-N'-[(substituted phenyl)methylidene]acetohydrazide) derivatives (Formula 2a-t), which are the molecules that are the subject of the invention on the cyclooxygenase enzymes COX-1 and COX-2.

As can be clearly seen from Table 2, the molecules that are the subject of the invention show similar/close effects to the molecules available on the market with proven selectivity and effectiveness.

Table 3. Cyclooxygenase (COX) IC50 values of active ingredients.

Some studies have been conducted to analyse the effects of 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3- yl]sulfanyl]acetohydrazide (Formula 1 ) and 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3-yl]sulfanyl]-N'- [(substituted phenyl)methylidene]acetohydrazide) derivatives (Formula 2a-t), which are the molecules that are the subject of the invention, on the MCF-7 breast cancer cell line in humans. "Dulbecco's modified Eagle medium I High Glucose" (DMEM/High, Gibco 41966) was used for the propagation of the HEK293 and MCF-7 cell lines used in the experiments. DMEM/high glucose content contains pyruvate, L-Glutamine and 4.5g/L D-glucose. The medium to be used in the experiments was prepared by adding 10% (v/v) fetal bovine serum (FBS, Gibco) and 1 % (v/v) antibiotic-antimycotic solution (Gibco) to the sterile media solution. Additionally, cells were obtained from ATCC. After cell opening, the cell culture flask was placed in the 37°C incubator. 24 hours after cell thawing, the viability of the cells in the cell culture flask was checked according to their adhesion status with an inverted microscope. When the cells proliferated enough to cover the surface by 80 percent, passage procedures were carried out.

In the preparation of subcultures of the cells, the medium in the 25 cm ² cell culture dish was first removed and washed with 5 ml of "Dulbecco Phosphate Buffered Saline" (D-PBS). After the washing process, D-PBS was removed from the medium. To ensure that the cells were lifted off the surface of the culture vessel, 1 mL Trypsin/EDTA was added and the culture vessel was placed in the incubator, and the process was physically and slowly tapping the edge of the culture vessel by hand to separate the cells from the surface and each other. The complete removal of the cells from the surface of the culture dish was checked with an inverted microscope. After the separation process was completed, 5 ml of the medium was placed into the culture container and pipetted to terminate the trypsin activity. Cell counting was done with a hemocytometer (Neubauer slide). Cells were placed in a new culture dish and the top was made up to 7 ml with fresh medium. The cells were placed in a 37°C incubator (Thermo Scientific, Heracell™ 1501) providing more than 90% relative humidity and 5% CO2, and this process was repeated two or three times a week for cell continuity. Subculturing of the cells was carried out in a sterile cabinet with laminar air flow (Heal Force HFsafe-1200 version 2.01 ).

After preparing subcultures of HEK293 and MCF-7 cells, cytotoxicity analyses of 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H- 1 ,2,4-triazol-3-yl]sulfanyl]acetohydrazide (Formula 1 ) and 2-[[4-(4-Fluorophenyl)-5-(furan-2-yl)-4H-1 ,2,4-triazol-3- yl]sulfanyl]-N'-[(substituted phenyl)methylidene]acetohydrazide) derivatives (Formula 2a-t), which are the molecules that are the subject of the invention, were also performed. In this method, first of all, by means of the mitochondrial dehydrogenase enzyme activity in living cells, the tetrazolium ring of the MTT [3-(4,5-dimethylthiazole-2,5- diphenyltetrazolium)] molecule was broken and the MTT molecule was transformed into water-insoluble formazan crystals. After the formed formazan crystals were dissolved with the help of dimethyl sulfoxide (DMSO), absorbance was measured in a spectrophorometer. For cytotoxicity analysis, the following steps were performed respectively. First of all, 200 pl of cell suspensions of 5.104 cells/well were placed in each well of the 96-well culture dishes. The culture vessel was incubated in an oven at 37°C providing 5% carbon dioxide (CO2). The medium on the cells attached to the surface of the wells was removed at the end of the 24 ^th hour. First of all, the viability of the media containing 50 pM substance was analysed, and then the effects of different concentrations were tested. 200 pl each of the medium containing different concentrations of agent was applied to the experimental group cells, and 200 pl of only the medium (DMEM/High, Gibco 41966) was applied to the control group, and they were kept in the incubator (37°C, 5% CO2) and the medium on the cells was removed at the end of the 24-hour incubation period. Subsequently, 30 pl of MTT stock solution (5mg/ml, prepared in PBS) was added and kept in the incubator for 4 hours. 150pl isopropanol was added to dissolve the formazan crystals and the culture container was shaken in a shaker (Heidolph Unimax 1010) at room temperature for solution homogeneity. In the next step, the absorbance of the resulting coloured product at 570 nm wavelength was measured on a microplate reader (Spektramax I3) and the results were evaluated according to the equation below. Experiments were carried out in three repetitions and in three separate periods.

In order to test the COX-1 and COX-2 enzyme activities of the synthesized molecules, the substances were dissolved in DMSO, then the effect of DMSO was eliminated and the DMSO ratio in the solutions was reduced to below 1 % with the buffer solution included in the kit. It was tested whether the synthesised substances inhibit COX-1 (sheep recombinant) and COX-2 (human recombinant). In this experiment, the reaction between PGG2 and 10-acetyl-3,7- dihydroxyphenoxazine (ADHP) produces the highly fluorescent compound resorufin. The %inhibition values were calculated by measuring the fluorescence resulting from the experiment. The enzyme inhibition experiment was carried out in accordance with the kit (COX Fluorescent Inhibitor Screening Assay Kit (Cayman, 700100)) manufacturer's instructions and enzyme inhibitions of the substances were measured in a multimode microplate reader (Spektramax i3) with an excitation wavelength of 530-540 nm and an emission wavelength of 585-595 nm. Enzyme inhibition of the substances at a concentration of 10 pM was evaluated by comparing them with standards.

All experiments were performed at three different times and in at least three replicates. Statistical data were evaluated using GraphPad Prism software version 7.02 (La Jolla, California, USA). Differences between groups were evaluated using one-way analysis of variance (ANOVA) and Dunnet's multiple comparison test, (at least P< 0.05).

Viability tests to determine the cytotoxic effects of the agents used in the study on MCF-7 and HEK293 cells were performed in 3x3 replicates. The control group in the experiments was determined as the group in which no agent was applied and was considered 100% alive. One-way ANOVA test was applied to the data obtained to analyse the consistency between repetitions within the experimental groups. The significance of the groups compared to the control group was evaluated with the Dunnett test and Tukey test. As a result of literature searches, the application concentrations and times of the agents were decided. The concentration values (IC50) of the agents applied to the cells that halved the cell viability were calculated. To calculate the IC50 value of the agents, the sigmoidal dose response variable slope was calculated using the GraphPad Prism® 7.0 program.

In order to determine the cytotoxic effects of substances on MCF-7 and HEK293 cells, media containing substances at different concentrations (50-0.1 pM) were added to 24-hour cultures. The medium of the control group, to which no substance was applied, was also refreshed and data was obtained by performing the MTT test after the 24-hour incubation period. The % viability results of the experiments where 50 pM substance was applied are shown in Figure 1 and Figure 2. IC50 results of the substances are shown in Table 1 .

In order to test the inhibition effects of the molecules that are the subject of the invention on COX-1 and COX-2 enzymes, procedures were carried out as recommended by the manufacturer. For this purpose, first the enzymes and substances were incubated, and then 200 pl of substance-enzyme cocktail containing 20 pM substance was placed in 96-well plates. Enzyme activity measured for each substance was carried out in triplicate and at three different times. One-way ANOVA test was applied to analyse the consistency between repetitions of experimental groups. The significance of the groups compared to the standard inhibitors group (SC-560 for COX-1 , DuP-697, celecoxib and indomethacin for COX -2) was evaluated with Dunnett's test. COX-1 and COX-2 enzyme inhibitions of the substances at 20 pM concentration are shown in Table 2, and IC50 values are shown in Table 3. All values show statistical significance (at least P<0.05).

References

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