STUDY OF THE BIOLOGICAL ACTIVITY OF 2-CHLORO-N-TOLYLACETAMIDE AND N-(TOLYL)-Α-PICOLINAMIDES OBTAINED BY N-ACYLATE OF TOLUIDINE ISOMERS WITH CHLOROACETYL CHLORIDE AND PICOLINE ACID IN THE PASS ONLINE PROGRAM

ABSTRACT

The biological activity of 2-chloro-N-tolylacetamides synthesized by the N-chloroacetylation reaction of toluidine isomers with chloroacetyl chloride and N-(tolyl)-α-picolinamides synthesized by the N-acylation reaction with picolinic acid was studied in the PASS online program [https://www.way2drug.com/PassOnline/index.php]. The effect of the change of the acyl group in the isomers 2-chloro-N-tolylacetamide and N-(tolyl)-α-picolinamides on the biological activity of the amide derivatives was examined and compared. The cytotoxic effect of N-(tolyl)-α-picolinamide isomers was observed in vitro against epithelial cervical cancer, breast adenocarcinoma, and laryngeal adenocarcinoma in laboratory conditions.

АННОТАЦИЯ

Биологическая активность 2-хлор-N-толилацетамидов, синтезированных реакцией N-хлорацетилирования изомеров толуидина с хлорацетилхлоридом, и N-(толил)-α-пиколинамидов, синтезированных реакцией N-ацилирования с пиколиновой кислотой, изучалась в онлайн-программе PASS [https://www.way2drug.com/PassOnline/index.php]. Было изучено и сравнено влияние изменения ацильной группы в изомерах 2-хлор-N-толилацетамида и N-(толил)-α-пиколинамидов на биологическую активность производных амида. Цитотоксический эффект изомеров N-(толил)-α-пиколинамида наблюдался in vitro против эпителиального рака шейки матки, аденокарциномы молочной железы и аденокарциномы гортани в лабораторных условиях.

Keywords: acyl group, 2-chloro-N-tolylacetamide, PASS online program, N-(tolyl)-α-picolinamide, Pa and Pi values.

Ключевые слова: ацилная группа, oнлайн-программа PASS, N-(толил)-α-пиколинамид, значения Pa и Pi, 2-хлор-N-толилацетамид.

Introduction. Today, the development of new, more effective, and safe drugs is one of the priority tasks of organic chemistry, and in the production of such drugs, the PASS, GUSAR and PharmaExpert programs are widely used, which allow predicting various types of biological activity with high accuracy based on the structural formulas of compounds belonging to various chemical classes. Based on these programs, the initial approximate primary biological activities of organic compounds are revealed and based on the primary results obtained  based on the program, the biological activities of substances are studied in practice. Synthesis based on the analysis of theoretical studies avoids the waste of time and unnecessary reagents.

Review of relevant literature. One of the most important properties of chemical compounds is their biological activity, the presence of which allows or, conversely, prevents the practical application of a chemical compound in medicine, cosmetics, food industry, agricultural chemistry, and other fields [1]. The functional possibilities of amides attract great attention in current chemical and industrial research not only due to their broad biological and pharmacological activity, but also because they are versatile reagents for the synthesis of various useful molecules. When reviewing the literature,  more than 50% of known drugs contain amide units. Among them, picolinamide-based anticancer drugs such as sorafenib and regorafenib (multikinase inhibitors), 2-anilinobenzothiazole derivatives (BRAFV600E and C-RAF inhibitor), and BLZ-945 (FMS inhibitor) contain N-methylpicolinamide and are its main components [2-6]. Nowadays, it is important to develop effective methods for the synthesis of amide-linked compounds from carboxylic acids and their halogen anhydrides. In this regard, convenient methods for the synthesis of amides based on the reactions of aniline and some substituted anilines with chloroacetyl chloride and picolinic acid have been developed, and the structure of the synthesized amide derivatives has been confirmed by IR, 1H NMR, 13C NMR spectroscopy, mass spectrometry and X-ray structural analysis [7-9]. Preliminary estimates of the primary biological activities of the obtained amide derivatives were studied in the PASS online program, and preliminary conclusions were drawn.

Research methodology. In this article, the biological activities of 2-chloro-N-tolylacetamides synthesized by the N-chloroacetylation reaction of toluidine isomers with chloroacetyl chloride and N-(tolyl)-α-picolinamides synthesized by the N-acylation reaction with picolinic acid were studied using the PASS online program [https://www.way2drug.com/PassOnline/index.php] and the influence of the structure of the amides on the biological activity was compared.

The studied amide derivatives were synthesized by the acylation reactions of toluidine isomers with chloroacetyl chloride and picolinic acid. Three different isomeric amides were synthesized by the reaction of toluidine isomers with picolinic acid.

Isomeric amides of three different structures were also synthesized from the reaction of toluidine isomers with chloroacetyl chloride.

The structure of the synthesized amide compounds was confirmed by IR, 1H NMR, 13C NMR spectroscopy, and Mass spectrometric methods [11]. Considering the structure of the synthesized amide compounds,  N-acyl products were formed in the presence of various acylating reagents, and the structure of the acyl group may affect the biological activity of the substance.

Analysis and results. Initially, the predicted biological activities of 2-chloro-N-tolylacetamide isomers and N-(tolyl)-α-picolinamides against some diseases were obtained and compared using the PASS online program.

Table 1.

Predicted biological activities of 2-chloro-N-tolyl acetamide and N-(tolyl)-α-picolinamide isomers against some diseases based on the program

The results of the table show that the biological properties of 2-Chloro-N-tolyl acetamide and N-(tolyl)-α-picolinamide isomers also changed with the change of the acyl group. That is, the change of the acyl group can cause the amide products to show biological activity against different types of casas. In the isomeric amides, biological activity was observed against the same type of casas. Only the Pa and Pi values ​​of the observed activity differ slightly from each other. In the 2-Chloro-N-tolyl acetamide isomers, 2-chloro-N-o-tolyl acetamide, 2-chloro-N-m-tolyl acetamide, and 2-chloro-N-p-tolyl acetamide, an increase in the Pa value was observed, which shows that as the substituent moves from the ortho- to the p- position, the biological activity index also increases. This pattern was not observed for N-(tolyl)-α-picolinamide isomers.

Based on the PASS online program, along with the results of the activity of chemical compounds against certain diseases, it is possible to obtain estimates of the side and toxic effects of a chemical compound on the body, and preliminary data on the toxic effects of the studied compound were obtained [https://www.way2drug.com/passonline] (Table 2).

Table 2.

Toxic effects of 2-Chloro-N-tolyl acetamide and N-(tolyl)-α-picolinamide isomers on the body

Analysis of the results shows that both 2-Chloro-N-tolyl acetamide and N-(tolyl)-α-picolinamide isomers cause cutaneous effects such as Cramp syndrome, peptic ulcer, aphthous ulcers, and Neutrophilic dermatosis (Sweet syndrome). N-(tolyl)-α-picolinamide isomers have less harmful effects than 2-Chloro-N-tolyl acetamide isomers.

The cytotoxic effect of the synthesized N-(tolyl)-α-picolinamide isomers was studied in vitro using the MTT method on 3 cancer cell lines (cervical epithelial carcinoma HeLa, mammary adenocarcinoma HBL-100 (ATCC NTB 124) and laryngeal adenocarcinoma HEp-2 (ATCC: CCL-23)). The substances were dissolved in DMSO (0.8% by volume) and added to the cells at concentrations of 50 and 100 μM.

Table 3.

Cytotoxic effects of N-(tolyl)-α-picolinamide isomers

The results of the test showed that none of the substances showed a pronounced cytotoxicity (IC50 value >100 μM) comparable to the effect of the reference drug in vitro. Also, none of the tested substances showed more than 10% inhibitory activity at a concentration of 50 μM in 3 cancer cell lines. N-(tolyl)-α-picolinamide isomers at a concentration of 100 μM showed a slight cytotoxicity in the HeLa cervical epithelial carcinoma cancer cell line - 24.1%, 27.3% and 26.7% cell growth inhibition, respectively. Thus, the studied N-(tolyl)-α-picolinamide isomers do not have cytotoxic activity in vitro against epithelial cervical cancer, breast adenocarcinoma and laryngeal adenocarcinoma.

Conclusions and suggestions. According to the results of the table, when comparing the biological activities of 2-Chloro-N-tolyl acetamide and N-(tolyl)-α-picolinamide isomers with each other, dramatic changes in biological activities can be observed with the change of the acyl group in the amide derivatives. The position of the methyl (-CH3) group in the benzene ring in the molecule of 2-Chloro-N-tolyl acetamide and N-(tolyl)-α-picolinamide isomers has almost no effect on the biological activity of the N-(tolyl)-α-picolinamide molecule, and their biological activities are similar regardless of the position of the methyl group in the ring. In 2-Chloro-N-tolyl acetamide isomers, the Pa and Pi values differ depending on the position of the methyl (-CH3) group in the benzene ring. The results obtained in the PASS online program serve as the basis for studying the biological activities of substances in practice. N-(tolyl)-α-picolinamide isomers do not have cytotoxic activity against epithelial cervical cancer, breast adenocarcinoma, and laryngeal adenocarcinoma in vitro under laboratory conditions.

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