UDC 547.792'854.4-145.55:615.276.015.11

A. S. Gotsulya, A. V. Samko, O. I. Panasenko

Anti-inflammatory activity of 2-(5-((theophylline-7'-yl)methyl)-4-R-4H-1,2,4-triazole-3-ylthio)acetic acid salts

Zaporozhye State Medical University

Abstract. Conducted the research of the inflammatory activity of the salts of 2-(5-((theophylline-7'-yl)methyl)-4-R-4Н-1,2,4-triazole-3-ylthio)acetate acid on the model of acute inflammation "formalin-induced edema of the extremities" of rats. Inflammatory reaction was modeled using 2,0% aqueous formalin solution. As a reference-preparation used sodium diclofenac. The research results have been processed by modern statistical methods of analysis. As the research results shows, the most active are 2-(5-((theophylline-7'-yl)methyl)-4-R-4Н-1,2,4-triazole-3-ylthio)аcetate acid and it’s diethyletanolammonium salt. Among the studied substances should also be noted a piperidine 2-(5-((theophylline-7'-yl)methyl)-4-phenyl-4Н-1,2,4-triazole-3-ylthio)acetate, which shows a high anti-inflammatory activity.

Key words: triazoles, theophylline, anti-inflammatory agents

Entry. The most common and frequently used in a number of diseases group of drugs NSAIDs are used to relieve inflammation or as an analgesic. The most active anti-inflammatory effect possessed by those NSAIDs non-selectively inhibit the cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2), disrupting the synthesis of prostaglandins and tromboksanov and, unfortunately, have a number of unwanted effects associated with ulcerogenic action of drugs. It is known that among guut NSAIDs selective COX-2 anti-inflammatory effect is much lower, but almost no side effects on the gastrointestinal mucosa, but leaves open the question of the use of selective COX-2 inhibitors in patients with risk of cardiovascular and cerebrovascular pathology.

As well as anti-inflammatory drugs myristoleate GCS, mechanism of action of which is based on the suppression of the transcription of the genes encoding the synthesis of proinflammatory proteins. The severity of the complications of GKS based on the level of their application and duration of use. Most often with long-term use of corticosteroids objnum effect is the development of candidiasis of the oral cavity, and one of the main complications of GCS is the occurrence of osteoporosis.

Thus, it is important to search for highly effective medicines, which could exhibit pronounced anti-inflammatory effect, but have a smaller segment of side effects. [1, 2, 3].

The aim of this work is to evaluate anti-inflammatory activity of salts of 2-(5-((theophylline-7'-yl)methyl)-4-R-4H-1,2,4-triazole-3-ylthio)acetic acid model of acute inflammation "formalin-induced edema of the limbs" of laboratory rats.

Materials and methods research. For the study was selected of 14 salts of 2-(5-((theophylline-7'-yl)-methyl)-4-phenyl-4H-1,2,4-triazole-3-ylthio)-acetic acid (tab.1), was synthesized at the Department of Toxicological and inorganic chemistry of Zaporozhye State Medical University. All experiments were conducted on intact white nonlinear rats of both sexes weighing from 190 to 260 grams for 7 animals in each group.

To achieve the purpose, an aqueous solution or suspension of the studied salts, stable tween-80 for water-insoluble compounds (at a dose of 1/10 of LD₅₀ per 1 ml of solution/suspension or 100 g. animals), was administered per os 30 min before the introduction of flogogen. Modeling the inflammatory response conducted by introducing as flogogen of 2.0% aqueous formalin solution [5]. Formalin was injected in a volume of 0.1 ml sub plantar in one of the hind limbs. Evaluation of anti-inflammatory activity was performed after 2 and 4 hours after administration of formalin by measuring volume of limbs volumetrically method. The limb volume expressed in cm³. As a reference drug used was diclofenac sodium (2,5% solution, ampoules of 3 ml, "Darnitsa", Ukraine), which was administered at a dose of 8 mg/kg [5].

The research results are processed by statistical methods of the analysis using, in particular, the standard software package of Microsoft Office 2016 (Microsoft Excel) and "STATISTICA^® for Windows 6.0". Calculated average arithmetic (M) and standard error (±m). The reliability of intergroup differences according to the experimental data was established using a student's t-test. Used 3 the level of statistical significance of research results, p<0.05; p<0.01; p<0,001 [4, 6, 8]

Table 1

Salts of 2-(5-((theophylline-7'-yl)methyl)-4-R-4H-1,2,4-triazole-3-ylthio)-acetic acid

№	R	X^+	№	R	X^+
1	СН3	Na^+	8	С6Н5	½ Ca^2+
2	СН3		9	С6Н5	½ Zn^2+
3	С2Н5		11	С6Н5
4	С2Н5		10	С6Н5
5	С2Н5		12	С6Н5
6	С2Н5		13	С6Н5
7	С6Н5	Н^+	14	С6Н5

Table 2

The anti-inflammatory activity of the salts of 2-(5-((theophylline-7'-yl)methyl)-4-R-4Н-1,2,4-triazole-3-ylthio)acetate acid

№	Compound/ group	Dose of compound mg/kg	The volume of paws of rats after 2 h, cm^3	Anti-inflammatory activity, %	· The statistical significance of the results		The volume of paws of rats after 4 h, cm^3	Anti-inflammatory activity, %	The statistical significance of the results
					in relation to the control	in relation to diclofenac			in relation to the control	in relation to diclofenac
1	Control of phus. sol.	-	0,46±0,027	0	-	р = 0,006	1,33±0,051	0	-	р=0,00027
2	Ddiclofenac	8	0,34±0,015	25,36	р = 0,006	-	0,81±0,058	38,78	р=0,00027	-
3	2	45	0,19±0,019	59,06	р = 7,39*10^-5	р = 0,000357	1,29±0,079	7,52	р=0,32	р=0,0038
4	6	46	0,19±0,023	59,38	р=1,2*10^-4	р = 0,00081	1,09±0,076	18,15	р=0,032	р=0,023
5	10	55	0,47±0,043	-3,12	р = 0,78	р = 0,02206	1,11±0,062	16,76	р=0,027	р=0,0104
6	14	53	0,38±0,028	17,19	р=0,08	р = 0,264	1,20±0,082	9,99	р=0,0066	р=0,212
7	Control	-	0,39±0,039	0	-	-	1,35±0,063	0	-	-
8	1	121	0,72±0,044	-83,33	р=0,0008	р = 7,81*10^-5	1,26±0,099	6,36	р=0,4868	р=0,0057
9	3	45	0,63±0,027	-61,84	р=0,0013	р = 2,9*10^-5	1,21±0,067	10,59	р=0,1621	р=0,0030
10	7	105	0,19±0,036	51,99	р=0,006	р = 0,0058	0,80±0,175	40,57	р=0,0214	р=0,946
11	13	50	0,21±0,027	46,41	р=0,00627	р = 0,00389	0,92±0,112	32,10	р=0,0116	р=0,456
12	Control	-	0,37±0,028	0	-	-	1,38±0,091	0	-	-
13	5	49	0,57±0,027	-53,26	р=0,0013	р = 0,000126	1,20±0,083	13,43	р=0,1756	р=0,0068
14	11	50	0,20±0,039	46,74	р=0,00819	р = 0,011302	1,13±0,063	18,08	р=0,0585	р=0,0073
15	Control	-	0,40±0,18	0	-	-	1,14±0,091	0	-	-
16	4	55	0,35±0,030	10,47	р=0,27	р = 0,67822	1,31±0,097	-15,16	р=0,2364	р=0,00312
17	8	115	0,16±0,009	60,65	р=7,35*10^-6	р = 1,41*10^-5	0,80±0,134	29,70	р=0,0749	р=0,932103
18	9	157	0,50±0,033	-27,44	р=0,23	р = 0,00235	1,37±0,079	-20,05	р=0,1018	р=0,000777
19	12	53	0,13±0,016	67,51	р=1,12*10^-5	р = 2,5*10^-5	0,62±0,080	45,74	р=0,00362	р=0,08815

Results and their discussion. The results of studies given in table 1 found that 2-(5-((theophylline-7'-yl)methyl)-4-R-4H-1,2,4-triazole-3-ylthio)acetic acid and its salt diethylethanolamine exceed power of the anti-inflammatory action of the reference drug as a 2 (26,63 % - 42,15 %, respectively, P ≤ 0,01) and 4 hours (by 1,79 % and of 6,96 %, respectively, P ≤ 0,05). Also it should also be noted piperidine 2-(5-((theophylline-7'-yl)methyl)-4-phenyl-4H-1,2,4-triazole-3-ylthio)-acetate, which showed high anti-inflammatory effect, the strength of which is close to the action of diclofenac sodium (6,68% less active P ≤ 0,05). 4 Hours connection 2, 5, 6, 8, 10, 11, 13 and 14 also showed a slight anti-inflammatory effect (in the range of 6,36 % - 32,10 %). 2 hours after the introduction of flogogen reduced the level of edema 7 connections more efficiently than the standard comparison. It was found that the presence of the cation diethylethanolamine in the molecule 2-(5-((theophylline-7'-yl)methyl)-4-R-4H-1,2,4-triazole-3-ylthio)acetate contributes to the identification of the most strong anti-inflammatory action. The lack of cation diethylethanolamine in the molecule 2-(5-((theophylline-7'-yl)methyl)-4-R-4H-1,2,4-triazole-3-ylthio)acetate acid was slightly reduced performance. Introduction cation morpholine to the synthon reduces the anti-inflammatory effect obtained salt is less effective than the reference drug.

Conclusions

1. According to the results of the pharmacological screening of salts of 2-(5-((theophylline-7'-yl)methyl)-4-R-4H-1,2,4-triazole-3-ylthio)acetic acid revealed a number of compounds with anti-inflammatory activity, which approach or exceed the activity of the reference drug.

2. Diethylethanolamine 2-(5-((theophylline-7'-yl)methyl)-4-R-4H-1,2,4-triazole-3-ylthio)acetate was the most active compound: when the application of reduced manifestations formalin edema within 4 hours by 45,74 %, which is 6,96% more active diclofenac (38,78 %, p = 0,00027).

Literature

1. Иманаева А. Я. Изучение противовоспалительной активности липоевой кислоты на модели адъювант-индуцированного артрита крыс / А. Я. Иманаева, Л. Н. Залялютдинова, Д. Э. Цыплаков // Biological sciences. Fundamental research. – 2013. – №6. – P. 596.

2. Димефосфон проявляет противовоспалительную и антиоксидантную активность на модели хронического аутоиммунного воспаления / И. Х. Валеева, Л. Е. Заганшина, В. Н. Хазиахметова, А. Ф. Титаренко // Экспериментальная и клиническая фармакология. – 2011, №3. – С.13 – 16.

3. Калитник А. А. Противовоспалительная активность хитозана и его низкомолекулярного производного / А. А. Калитник, П. А. Марков, А. В. Володько // Медицинский академический журнал. – 2012. – Т. 12, №3. – С. 60.

4. Лапач С. Н. Статистические методы в медико-биологических исследованиях с использованием Excel / С. Н. Лапач, А. В. Чубенко, П. Н. Бабич. – [2 изд.], перераб. и доп. – К. : Морион, 2001. – 408 с.

5. Методичні вказівки по доклінічному вивченню лікарських засобів: Під ред. А. В. Стефанова. – К., 2001. – 567 с.

6. Реброва О. Ю. Статистический анализ медицинских данных. Применение пакета прикладних программ STATISTICA / О. Ю. Реброва. – М.: МедиаСфера, 2002. – 312 с.

7. Руководство по экспериментальному (доклиническому) излучению новых фармакологических веществ / Под общей редакцией чл.-корр. РАМН проф. Р. У. Хабриева. – 2-изд., перераб. и доп. – М.: Медицина, 2005. – 832 с.

8. Сернов Л. Н. Элементы экспериментальной фармакологии / Л. Н. Сернов, В. В. Гацура. – М.: ВНЦ БАВ, 2000. – 352 с.