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Synthesis and antioxidant properties of new (2,4- and 3,4-dimethoxyphenyl)-1,2,4-triazoles
expand article infoDmitro V. Dovbnya‡, Andriy H. Kaplaushenko‡, Yuliia S. Frolova‡, Evheniy S. Pruglo‡
‡ Zaporizhzhіa State Medical University, Zaporizhzhia, Ukraine

Corresponding author: Yuliia S. Frolova ( yuliia_hulina@ukr.net )

Academic editor: Plamen Peikov
© 2022 Dmitro V. Dovbnya, Andriy H. Kaplaushenko, Yuliia S. Frolova, Evheniy S. Pruglo.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation: Dovbnya DV, Kaplaushenko AH, Frolova YS, Pruglo ES (2022) Synthesis and antioxidant properties of new (2,4- and 3,4-dimethoxyphenyl)-1,2,4-triazoles. Pharmacia 69(1): 135-142. https://doi.org/10.3897/pharmacia.69.e74107
Open Access

Abstract

The purpose of the work is to develop preparative methods for the synthesis of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles, to investigate the reaction of acid hydrolysis, to receive the physical-chemical properties of the synthesized compounds, and to study antioxidant activity of new compounds. Preparative methods for the synthesis of (5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles have been developed for which studied the reaction of acid hydrolysis, resulting in the production of carboxylic acids. The structure of the obtained substances was confirmed by modern physical-chemical methods. The antioxidant activity of the synthesized compounds was evaluated in vitro by the method of the non-enzymatic initiation of BOD with salts of iron (II).

Keywords

1, 2, 4-triazole, synthesis, biological activity, antioxidant activity

Introduction

In the 21st century, pharmacy is very popular and advanced science all over the world. There are many medicines with different pharmacological activities, but not even enough. Therefore, the search, synthesis, and implementation of new drugs with a wide range of biological activity and low toxicity is currently an urgent task of pharmacy.

Analysis of the current literature (Kaplaushenko 2008, 2013, 2015; Samelyuk and Kaplaushenko 2014, 2015; Kaplaushenko et al. 2016; Samelyuk 2016; Hulina and Kaplaushenko 2018; Sahu et al. 2018; Safonov 2020) indicates the prospect of finding biologically active substances among 2-((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)acetonitriles. The works of ZSMU scientific school based on dissertations (Kaplaushenko 2008; Samelyuk 2016) and articles (Kaplaushenko 2013; Hulina and Kaplaushenko 2018) show that the discussed class of compounds serves as a basis for the creation of potential original drugs, with some already being actively used in medicine.

It is impossible to neglect the work of domestic authors (Samelyuk and Kaplaushenko 2014, 2015; Kaplaushenko et al. 2016; Sahu et al. 2018; Shcherbyna et al. 2019; Safonov 2020), which presents the results of the pharmacological activity of 5-R-1,2,4-triazole-3-ylthiol. But the pharmacological activity in a number of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles has not been sufficiently studied. Therefore, synthesis, study of physical-chemical and biological properties of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano- , benzo)nitriles are scientific novelty, theoretical and practical importance.

To obtain ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles (compounds 2.1–2.11, Fig. 1) the interaction between the corresponding halogennitriles (chloroacetonitrile, 3-chloropropanonitrile, 4-chlorobutanonitrile, 2-chlorobenzonitrile, 4-amino-2-chlorobenzonitrile, 3-(chloromethyl)benzonitrile) with 5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-thione (compounds 1.1, 1.2, Fig. 1) in an alkaline alcohol medium were used.

Figure 1. 

Scheme of the synthesis of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles (compounds 2.1–2.11).

The synthesized nitriles (2.1–2.11) (Table 1) are crystalline substances of yellow (2.1, 2.7, 2.11), white (2.2, 2.10), orange (2.3, 2.9), brown (2.4, 2.5, 2.8), black (2.6) colors, slightly soluble in water, soluble in alkaline solutions, as well as in organic solvents and mineral acids. For the analysis substances (2.1–2.11) were recrystallized from ethanol.

It is known that hydrolysis of nitriles can be performed by two methods: alkaline and acid hydrolysis. Considering the work of scientists (Kaplaushenko 2013; Samelyuk and Kaplaushenko 2015; Shcherbyna 2019), it can be concluded that in acid hydrolysis the target product is obtained with a higher percentage of yield. Therefore, we subsequently studied the reaction of acid hydrolysis for aceto(propano-, benzo)nitriles, resulting in acid.

The preparation of this class of compounds was carried out by the interaction of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, benzo)nitriles (compounds 2.1 , 2.7, 2.10, Fig. 2) with acid chloride, water was used as a solvent.

Figure 2. 

Scheme of the synthesis of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propanoic, benzoic)acids (compounds 2.12–2.14).

The obtained 2-((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)acetate(propanoic, benzoic)acids (Table 1) are brown (2.12), orange (2.13), or white (2.14) crystalline substances, soluble in water (heated), in alkali metal, and in organic solvents. For the analysis acids (2.12–2.14) were recrystallized from propanol-water 2:1.

Toxicity

At the first stage of the study of biological activity of the derivatives of 5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-thione the acute prediction of toxicity was performed with the program GUSAR-online. It was done to weed out potentially toxic substances as unpromising objects experimental pharmacological screening. Computer prediction of acute toxicity of the derivatives of 5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-thione were carried out according to structural formulas compounds in the online version of GUSAR-online.

The on-line prognosis was performed for 14 compounds derived from 5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-thione.

Antioxidant activity

The method of evaluation of AOA was used in the non-enzymatic initiation of BOD with salts of iron (II) (Pruglo 2017).

It has been chosen this method because it was the most accessible. With this method, antioxidant activity can be determined without animals. Also, the method of evaluation of AOA has made it possible to predict antihypoxic and antiischemic activity. For such drugs as Trifuzol, Avestim, Thiotriazolin, Thiometrizol, and many others, this research method was also used (Bushueva et al. 2017).

The egg lipoprotein suspension (ELS) was used as the substrate. ELS was prepared by homogenizing egg yolk with phosphate buffer (pH = 7.4). To the suspension was added the test compounds at a concentration of 10-3 mol / l. The free radical oxidation reaction is initiated by the addition of FeSO4 × 7H2O solution. The mixture was incubated for 60 min at 37 °C. The reaction was stopped with a 20% solution of trichloroacetic acid with trilon B. After centrifugation for 30 min. a solution of thiobarbituric acid (TBA) was added to the supernatant and boiled in a water bath for 60 minutes. The colored complex of TBA-active products (TBA – AP) is extracted with the addition of n-butanol. Spectrophotometry determines the concentration of TBK-AP. Antioxidant activity (in percent) is determined by the formula:

AOA=E0-E1E0×100%

where AOA – antioxidant activity, %

E0 – the optical density of the control solution;

E1 – the optical density of a solution containing the test compound (vitamin C).

((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles (compounds 2.1–2.11)

A mixture of 0.03 mol of 5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-thione (compounds 1.1–1.2) and 0.03 mol of sodium hydroxide solution in 50 ml of methanol, which was heated to complete dissolution of the corresponding thione (1.1, 1.2), 0.03 mol of the corresponding halogennitrile (chloroacetonitrile, 3-chloropropanitrile, 4-chlorobutanonitrile, 2-chlorobenzonitrile, 4-amino-2-chlorobenzonitrile, 3-chlorobenzonitrile) were added to the reaction mixture and were heated to a neutral medium.

The primary precipitate which had been formed of sodium chloride was filtered off. After complete cooling ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles (compounds 2.1–2.11) were filtered off, washed with diethyl ether and dried.

2-((5-(2,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)acetonitrile (2.1). Yield 98%, m.p.= 153–155 °C. Adsorption maxima in IR-spectra VC=N cycle = 1573 cm-1; VC≡N = 2247 cm-1; VO-CH3 = 2817 cm-1; VCH2s/as = 2843/2920 cm-1; VAr = 1485 cm-1; VC-S = 634 cm-1. 1H NMR (400 MHz, DMSO-d6) d 2.60–3.70 (2H, s, S-CH2); 3.82–3.89 (6H, d, O-CH3); 4.21(1H, s, CH); 6.67–7.83(3H, m, C6H3). Calcd for C12H12N4O2S %: C, 52.16; H, 4.38; N, 20.28; S, 11.60. Found %: C, 52.17; H, 4.39; N, 20.27; S, 11.59. m \ z + (+1 amp) 276,31.

3-((5-(2,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)propanenitrile (2.2). Yield 92%, m.p.= 133–135 °C. Adsorption maxima in IR-spectra VC=N cycle = 1505 cm-1; VC≡N = 2270 cm-1; VO-CH3 = 2825 cm-1; VCH2s/as = 2862/2935 cm-1; VAr = 1497 cm-1; VC-S = 622 cm-1. 1H NMR (400 MHz, DMSO-d6) d 2.78 (2H, m, CH2-CH2); 3.70 (2H, s, S-CH2); 3.83–3.90 (6H, d, O-CH3); 4.20(1H, s, CH); 6.67–7.49(3H, m, C6H3). Calcd for C13H14N4O2S %: C, 53.78; H, 4.86; N, 19.30; S, 11.04. Found %: C, 53.79; H, 4.85; N, 19.29; S, 11.05. m \ z + (+1 amp) 290,34.

4-((5-(2,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)butanenitrile (2.3). Yield 85%, m.p.= 123–125 °C. Adsorption maxima in IR-spectra VC=N cycle = 1596 cm-1; VC≡N = 2210 cm-1; VO-CH3 = 2827 cm-1; VCH2s/as = 2867/2922 cm-1; VAr = 1512 cm-1; VC−S = 583 cm-1. 1H NMR (400 MHz, DMSO-d6) d 1.85–2.76 (4H, m, CH2-CH2); 2.60–3.70 (2H, s, S-CH2); 3.84–3.90 (6H, d, O-CH3); 4.20(1H, s, CH); 6.67–7.81(3H, m, C6H3). Calcd for C14H16N4O2S %: C, 55.25; H, 5.30; N, 18.41; S, 10.53. Found %: C, 55.27; H, 5.32; N, 18.39; S, 10.51. m \ z + (+1 amp) 304,37.

2-((5-(2,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)benzonitrile (2.4). Yield 81%, m.p.= 95–97 °C. Adsorption maxima in IR-spectra VC=N cycle = 1530 cm-1; VC≡N = 2249 cm-1; VO-CH3 = 2825 cm-1; VCH2s/as = 2848/2926 cm-1; VAr = 1501 cm-1; VC−S = 678 cm-1. 1H NMR (400 MHz, DMSO-d6) d 3.83–3.89 (6H, d, O-CH3); 4.15(1H, s, CH); 6.67–7.81(3H, m, C6H3); 7.42–7.95(4H, m, C6H4). Calcd for C17H14N4O2S %: C, 60.34; H, 4.17; N, 16.56; S, 9.47. Found %: C, 60.35; H, 4.16; N, 16.55; S, 9.48. m \ z + (+1 amp) 338,39.

4-amino-2-((5-(2,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)benzonitrile (2.5). Yield 85%, m.p.= 115–117 °C. Adsorption maxima in IR-spectra VC=N cycle = 1579 cm-1; VC≡N = 2263 cm-1; VO-CH3 = 2821 cm-1; VCH2s/as = 2856/2919 cm-1; VAr = 1514 cm-1; VC−S = 608 cm-1. 1H NMR (400 MHz, DMSO-d6) d 3.80–3.85 (6H, d, O-CH3); 4.17(1H, s, CH); 5.29 (2H, s, NH2); 6.68–7.81(3H, m, C6H3); 7.42–7.95(3H, m, C6H3). Calcd for C17H15N5O2S %: C, 57.78; H, 4.28; N, 19.82; S, 9.07. Found %: C, 57.80; H, 4.30; N, 19.80; S, 9.05. m \ z + (+1 amp) 353,40.

2-((5-(3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)acetonitrile (2.6). Yield 98%, m.p.= 173–175 °C. Adsorption maxima in IR-spectra VC=N cycle = 1522 cm-1; VC≡N = 2235 cm-1; VO-CH3 = 2815 cm-1; VCH2s/as = 2847/2923 cm-1; VAr = 1482 cm-1; VC−S = 665 cm-1. 1H NMR (400 MHz, DMSO-d6) d 2.60–3.56 (2H, s, S-CH2); 3.82–3.84 (6H, d, O-CH3); 4.18(1H, s, CH); 6.93–7.45(3H, m, C6H3). Calcd for C12H12N4O2S %: C, 52.16; H, 4.38; N, 20.28; S, 11.60. Found %: C, 52.14; H, 4.36; N, 20.30; S, 11.62. m \ z + (+1 amp) 338,39. m \ z + (+1 amp) 276,31.

3-((5-(3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)propanenitrile (2.7). Yield 99%, m.p.= 176–178 °C. Adsorption maxima in IR-spectra VC=N cycle = 1537 cm-1; VC≡N = 2236 cm-1; VO-CH3 = 2827 cm-1; VCH2s/as = 2863/2930 cm-1; VAr = 1587 cm-1; VC−S = 679 cm-1. 1H NMR (400 MHz, DMSO-d6) d 2.78 (2H, m, CH2-CH2); 2.81 (2H, s, S-CH2); 3.83–3.85 (6H, d, O-CH3); 4.20(1H, s, CH); 6.98–7.49(3H, m, C6H3). Calcd for C13H14N4O2S %: C, 53.78; H, 4.86; N, 19.30; S, 11.04. Found %: C, 53.80; H, 4.84; N, 19.29; S, 11.05. m \ z + (+1 amp) 290,34.

4-((5-(3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)butanenitrile (2.8). Yield 66%, m.p.= 97–99 °C. Adsorption maxima in IR-spectra VC=N cycle = 1512 cm-1; VC≡N = 2243 cm-1; VO-CH3 = 2824 cm-1; VCH2s/as = 2845/2943 cm-1; VAr = 1488 cm-1; VC−S = 657 cm-1. 1H NMR (400 MHz, DMSO-d6) d 1.87–2.78 (4H, m, CH2-CH2); 2.83 (2H, s, S-CH2); 3.84–3.86 (6H, d, O-CH3); 4.22(1H, s, CH); 6.97–7.47(3H, m, C6H3). Calcd for C14H16N4O2S %: C, 55.25; H, 5.30; N, 18.41; S, 10.53. Found %: C, 55.26; H, 5.31; N, 18.40; S, 10.52. m \ z + (+1 amp) 304,37.

2-(((5-(3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)methyl)benzonitrile (2.9). Yield 56%, m.p.= 112–114 °C. Adsorption maxima in IR-spectra VC=N cycle = 1555 cm-1; VC≡N = 2231 cm-1; VO-CH3 = 2811 cm-1; VCH2s/as = 2840/2942 cm-1; VAr = 1505 cm-1; VC−S = 640 cm-1. 1H NMR (400 MHz, DMSO-d6) d 3.70 (2H, s, S-CH2); 3.83–3.85 (6H, d, O-CH3); 4.20(1H, s, CH); 6.98–7.49(3H, m, C6H3); 7.34–7.55(4H, m, C6H4). Calcd for C18H16N4O2S %: C, 61.35; H, 4.58; N, 15.90; S, 9.10. Found %: C, 61.37; H, 4.60; N, 15.88; S, 9.08. m \ z + (+1 amp) 338,39.

2-((5-(3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)benzonitrile (2.10). Yield 93%, m.p.= 93–95 °C. Adsorption maxima in IR-spectra VC=N cycle = 1538 cm-1; VC≡N = 2247 cm-1; VO-CH3 = 2816 cm-1; VCH2s/as = 2863/2928 cm-1; VAr = 1503 cm-1; VC−S = 701 cm-1. 1H NMR (400 MHz, DMSO-d6) d 3.87–3.89 (6H, d, O-CH3); 4.15(1H, s, CH); 6.67–7.95(4H, m, C6H4); 6.91–7.42(3H, m, C6H3). Calcd for C17H14N4O2S %: C, 60.34; H, 4.17; N, 16.56; S, 9.47. Found %: C, 60.32; H, 4.15; N, 16.58; S, 9.49. m \ z + (+1 amp) 352,41.

4-amino-2-((5-(3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)benzonitrile (2.11). Yield 94%, m.p.= 121–123 °C. Adsorption maxima in IR-spectra VC=N cycle = 1509 cm-1; VC≡N = 2252 cm-1; VO-CH3 = 2828 cm-1; VCH2s/as = 2861/2917 cm-1; VAr = 1511 cm-1; VC−S = 597 cm-1. 1H NMR (400 MHz, DMSO-d6) d 3.81–3.83 (6H, d, O-CH3); 4.13(1H, s, CH); 5.28 (2H, s, NH2); 6.37–7.54(3H, m, C6H3); 6.98–7.49(3H, m, C6H3). Calcd for C17H15N5O2S %: C, 57.78; H, 4.28; N, 19.82; S, 9.07. Found %: C, 57.77; H, 4.29; N, 19.83; S, 9.06. m \ z + (+1 amp) 353,40.

((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propanoic, benzoic)acid (compounds 2.12–2.14)

1 Mol of the corresponding nitrile (2.1, 2.7, 2.10), 65 ml of concentrated chloride acid were loaded into a glass of 250 ml and left at room temperature for 5 days for dissolved. Then 200 ml of water were added. The precipitates of the synthesized compounds were filtered off and air-dried.

2-((5-(2,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)acetic acid (2.12). Yield 82%, m.p.= 143–145 °C. Adsorption maxima in IR-spectra VC=N cycle = 1600 cm-1; VCH3COOH = 1698 cm-1; VO-CH3 = 2821 cm-1; VCH2s/as = 2843/2929 cm-1; VAr = 1614 cm-1; VC−S = 637 cm-1. 1H NMR (400 MHz, DMSO-d6) d 3.38 (2H, s, S-CH2); 3.83–3.90 (6H, d, O-CH3); 4.22(1H, s, CH); 6.76–7.49(3H, m, C6H3); 12.34(1H, s, COOH). Calcd for C12H13N3O4S %: C, 48.81; H, 4.44; N, 14.23; S, 10.86. Found %: C, 48.82; H, 4.45; N, 14.21; S, 10.86. m \ z + (+1 amp) 295,31.

3-((5-(3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)propanoic acid (2.13). Yield 44%, m.p.= 183–185 °C. Adsorption maxima in IR-spectra VC=N cycle = 1588 cm-1; VCH3COOH = 1760 cm-1; VO-CH3 = 2827 cm-1; VCH2s/as = 2848/2937 cm-1; VAr = 1605 cm-1; VC−S = 659 cm-1. 1H NMR (400 MHz, DMSO-d6) d 2.60 (2H, m, CH2-CH2); 2.71 (2H, s, S-CH2); 3.83–3.85 (6H, d, O-CH3); 4.20(1H, s, CH); 6.98–7.49(3H, m, C6H3); 12.17(1H, s, COOH). Calcd for C13H15N3O4S %: C, 50.48; H, 4.89; N, 13.58; S, 10.36. Found %: C, 50.47; H, 4.88; N, 13.60; S, 10.36. m \ z + (+1 amp) 309,34.

2-((5-(3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)benzoic acid (2.14). Yield 47%, m.p.= 187–189 °C. Adsorption maxima in IR-spectra VC=N cycle = 1593 cm-1; VCH3COOH = 1741 cm-1; VO-CH3 = 2818 cm-1; VCH2s/as = 2867/2922 cm-1; VAr = 1608 cm-1; VC−S = 692 cm-1. 1H NMR (400 MHz, DMSO-d6) d 3.85–3.87 (6H, d, O-CH3); 4.15(1H, s, CH); 6.98–7.49(3H, m, C6H3); 7.69–8.30(4H, m, C6H4); 12.75(1H, s, COOH). Calcd for C17H15N3O4S %: C, 57.13; H, 4.23; N, 11.76; S, 8.97. Found %: C, 57.15; H, 4.25; N, 11.74; S, 8.95. m \ z + (+1 amp) 357,38.

In the IR spectra of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles (compounds 2.1–2.11) there are absorption bands of vibrations of nitrile groups C=N at 2270–2210 cm-1, scissor strips at 2870–2840 and at 2950–2915 cm-1, methylene groups, as well as the bands inherent in aromatics and C = N-groups in the range of 1530–1505 and 1596–1550 cm-1 (Fig. 3).

Figure 3. 

IR spectra of 2-(((5-(3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)methyl)benzonitrile (2.9).

Absorption bands in the IR spectra of all synthesized acids (2.12–2.14) can be caused by the presence of –C=N- groups at 1600–1588 cm-1, absorption bands of the aromatic ring at 1614–1605 cm-1, in the IR spectra of acids there are CH2-COOH groups at 1760–1698 cm-1 (Kazitsyna 1979).

1H NMR spectra of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles (compounds 2.1–2.11) were characterized by the presence of multiplet signals of aromatic protons at 6.67–7.95 ppm, duplicate signals of protons of methoxy groups at 3.80–3.90 ppm. Singlet signals of the thiomethylene group at 2.60–3.70 ppm were also supported, which was confirming the passage of the alkylation reaction on the Sulfur atom (Samelyuk and Kaplaushenko 2014).

1H NMR spectra of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propanoic-, benzoic)acids (compounds 2.12–2.14) were characterized by extended singlets of protons of the carboxyl groups at 12.17–12.75 ppm, singlet signals of protons of the S-CH2 groups at 2.71–3.38 ppm, which was confirmed the passage of the alkylation reaction, as well as characteristic singlet signals of protons of the methoxy groups at 3.83–3.90 ppm (Kazitsyna 1979).

The identity of the synthesized compounds was proved by chromatographic mass spectrometry. However, only one peak corresponding to the molecular weight of the product of the interaction m \ z + (+1 amp) was detected (Figs 4, 5).

Figure 4. 

Mass spectra of 4-((5-(2,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)butanenitrile (2.3).

Figure 5. 

Mass spectra of 2-((5-(2,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)acetonitrile (2.1).

Toxicity

According to the results of the GUSAR-online, for derivatives of 5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-thione average lethal dose LD50 was when administered: intraperitoneally – from 273.0 to 865.0 mg / kg, intravenously – from 124.2 to 398.8 mg / kg, orally – from 348.1 to 1722.0 mg / kg and subcutaneously – from 586.0 to 2341.0 mg / kg (Table 1).

Table 1.
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Prediction of acute toxicity of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles using GUSAR-online prognosis.

Number compounds Rat IP LD50 (mg/kg) Rat IV LD50 (mg/kg) Rat Oral LD50 (mg/kg) Rat SC LD50 (mg/kg)
2.1 384,700 in AD 187,200 in AD 1420,000 out of AD 733,300 in AD
2.2 384,400 in AD 209,300 in AD 1650,000 out of AD 632,800 out of AD
2.3 409,900 out of AD 216,400 in AD 978,800 in AD 941,500 out of AD
2.4 446,300 in AD 164,800 in AD 1722,000 in AD 1742,000 out of AD
2.5 540,800 in AD 233,800 in AD 787,700 in AD 708,900 out of AD
2.6 259,800 in AD 166,200 in AD 348,100 in AD 636,400 in AD
2.7 391,000 out of AD 188,100 in AD 1348,000 out of AD 1377,000 in AD
2.8 398,000 out of AD 199,600 in AD 1701,000 out of AD 1544,000 out of AD
2.9 865,000 in AD 124,200 in AD 1195,000 in AD 2341,000 in AD
2.10 684,900 in AD 138,200 in AD 1268,000 in AD 1464,000 in AD
2.11 574,400 in AD 195,400 in AD 1000,000 in AD 1654,000 out of AD
2.12 840,300 in AD 335,800 in AD 1458,000 in AD 1929,000 in AD
2.13 273,000 in AD 398,800 in AD 1014,000 in AD 586,000 in AD
2.14 455,400 in AD 340,500 in AD 1235,000 in AD 1043,000 in AD

Antioxidant activity

For the study of antioxidant activity, ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles (2.1–2.11) were chosen, because acids did not give high values of this activity. This fact was confirmed by the scientists of our scientific school of ZSMU (Kaplaushenko 2008; Samelyuk 2016). ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propanoic, benzoic)acids (2.12–2.14) did not show high indicators of antioxidant activity, but their derivatives: salts, nitriles, ethers were quite active.

The results of the determination of AOA in model experiments under conditions Fe2+ -induced POL are presented in table 1. It can be seen: 3 compounds in varying degrees of expression can inhibit the generation of free radicals.

Moderate antioxidant activity among the studied of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles possessed in compounds 2.2, 2.10 which reduced the level TBC – AP by 12.07–13.55% (p < 0.001).

Compounds 2.1, 2.8 had high AOA, which reduced the TBC-AP content by 16.55–16.71% (p < 0.001).

The most pronounced AOA had methylammonium 3-((5-(3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)propanenitrile (2.7), which reduced the TBC-AP content by 29.03% (p < 0.001), but did not reach the level of the ascorbic reference drug by this ability acid by 7.94% (Table 2).

Table 2.
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Antioxidant activity of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propano-, butano-, benzo)nitriles on their derivatives in vitro at non-enzymatic initiation of VRO.

Compound Optical density (λ = 232 нм) M ± m (n = 7) АОА, %
Control 0.69±0.01 0
Vitamin C 0.43±0.0 36.97
2.1 0.57±0.01 16.55
2.2 0.6±0.01 12.07
2.3 0.69±0.01 -0.6
2.4 0.7±0.01 -5.55
2.5 0.78±0.01 -13.49
2.6 0.68±0.01 0.65
2.7 0.49±0.01 29.03
2.8 0.57±0.01 16.71
2.9 0.73±0.0 -5.55
2.10 0.59±0.01 13.55
2.11 0.68±0.01 0.44

In future research, we plan to carry out the synthesis of the derivatives of ((5-(2,4- and 3,4-dimethoxyphenyl)-3H-1,2,4-triazole-3-yl)thio)aceto(propanoic, benzoic)acids, such as salts and esters. As the analysis of the previous works of our scientific school of ZSMU (Kaplaushenko 2008; Samelyuk 2016) was shown that salts and esters were exhibited the highest indicators of antioxidant activity. Salts were showed high performance, but short-term activity. Esters had a longer-lasting effect but had not high enough.

Conclusions

  1. New ((5-(2,4- and 3,4-dimethoxyphenyl)-3 H-1,2,4-triazole-3-yl)thio)nitriles were synthesized.
  2. Preparative methods for the synthesis of ((5-(2,4- and 3,4-dimethoxyphenyl)-3 H-1,2,4-triazole-3-yl)thio)aceto(propanoic, benzoic) acids have been developed.
  3. The structure of the obtained compounds was confirmed by elemental analysis by IR spectroscopy, clear bands of oscillations of nitrile groups of CN at 2270–2210 cm -1 were revealed, which confirms the production of the target product and 1H NMR spectra, and their individuality by chromato-mass spectrometry.
  4. Pharmacological screening did not make it possible to identify compounds whose antioxidant activity exceeded that of ascorbic acid. Further optimization of the structure to improve their activities is currently in progress.

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