RU2806372C1 - Use of 3-methyl-5-oxo-n-phenyl-5h-indeno[1,2-c]pyridazine-4-carboxamide as agent with anti-inflammatory activity - Google Patents

Abstract

FIELD: organic chemistry; pharmaceuticals.

SUBSTANCE: invention relates to the use of biologically active compounds of N-aryl-3-methyl-5-oxo-5H-indeno[1,2-c]pyridazine-4-carboxamides class. The use of 3-methyl-5-oxo-N-phenyl-5H-indeno[1,2-c]pyridazine-4-carboxamide of formula (I) as an agent with anti-inflammatory activity is disclosed.

I

EFFECT: invention provides effective anti-inflammatory activity.

1 cl, 1 tbl, 2 ex

Description

The invention relates to the field of application of biologically active compounds of the class N -aryl-3-methyl-5-oxo -5H- indeno[1,2- c ]pyridazine-4-carboxamides, namely 3-methyl-5-oxo- N -phenyl - 5H- indeno[1,2- c ]pyridazine-4-carboxamide of formula ( I ):

I

possessing anti-inflammatory activity, which makes it possible to propose its use in pharmacology and medicine as an anti-inflammatory drug.

A method for the synthesis of N -aryl-3-methyl-5-oxo- 5H- indeno[1,2- c ]pyridazine-4-carboxamides is known from the prior art [An eco-friendly stereoselective synthesis of novel derivatives of indeno[1,2- b ]pyrrole and indeno[1,2- c ]pyridazine/ Vladimir L. Gein, Natalia V. Nosova, Andrei N. Yankin, Alina Y. Bazhina, Maksim V. Dmitriev // Polycyclic Aromatic Compounds. - 2021. - V. 41.- P. 540-552].

The anti-inflammatory activity of N -aryl-3-methyl-5-oxo -5H -indeno[1,2- c ]pyridazine-4-carboxamides has not been previously studied.

The objective of the invention is to search among indeno[1,2- c ]pyridazine-4-carboxamide derivatives for a compound exhibiting high anti-inflammatory activity with low toxicity.

The problem is solved by obtaining the compound 3-methyl-5-oxo- N -phenyl- 5H -indeno[1,2- c ]pyridazine-4-carboxamide ( I ), which exhibits high anti-inflammatory activity and low toxicity.

The claimed compound ( I ) is synthesized by the reaction of 3a,8b-dihydroxy-3-(1-hydroxyethylidene)-1-phenyl-1,3,3a,8b-tetrahydroindeno[1,2- b ]pyrrole-2,4-dione ( II ) with hydrazine hydrate by boiling the reagents in ethyl alcohol in the absence of a catalyst according to the scheme:

Scheme 1

I II

The synthesis of the starting 3a,8b-dihydroxy-3-(1-hydroxyethylidene)-1-phenyl-1,3,3a,8b-tetrahydroindeno[1,2- b ]pyrrole-2,4-dione ( II ) was carried out as previously described method [An eco-friendly stereoselective synthesis of novel derivatives of indeno[1,2- b ]pyrrole and indeno[1,2- c ]pyridazine/ Vladimir L. Gein, Natalia V. Nosova, Andrei N. Yankin, Alina Y. Bazhina, Maksim V. Dmitriev // Polycyclic Aromatic Compounds. - 2021. - V. 41. - P. 540-552].

The invention is illustrated by the following examples.

Example 1. Preparation of the claimed compound (I).

To a solution of 1 g (0.003 mol) 3a,8b-dihydroxy-3-(1-hydroxyethylidene)-1-phenyl-1,3,3a,8b-tetrahydroindeno[1,2- b ]pyrrole-2,4-dione ( II ) 0.0033 mol (10% excess) of hydrazine hydrate was added to 20 ml of ethyl alcohol. The reaction mixture was boiled for 15 minutes, then cooled, the precipitated crystals were filtered off and recrystallized from ethyl alcohol. Yield 73% (0.7 g), mp. 273-274°C (ethanol).

The claimed compound ( I ) is a yellow crystalline substance, soluble in dimethyl sulfoxide (DMSO), dimethylformamide (DMF), when heated - in ethyl and isopropyl alcohol, acetone, insoluble in water, stable during storage.

IR spectrum (Specord M-80 in KBr tablets), ν, cm ^-1 : 3277 (NH), 1728 (CO), 1654 (CON). ^1H NMR spectrum (Bruker AVANCE III HD 400 (400 and 100 MHz _{, respectively) in DMSO-d6} , internal standard - TMS), δ, ppm: 2.73 s (3H, _CH3 ), 7.20 t (1H, ArH, J 7.4 Hz), 7.42 t (2H, ArH, J 7.9 Hz), 7.67 d (2H, ArH, J 7.4 Hz), 7.71 d (1H, ArH, J 7.5 Hz), 7.83-7.92 m (2H, ArH), 8.16 d (1H, ArH, J 7.5 Hz), 10.66 s (1H, NH). Found, %: C 72.18; H 4.30; N 13.42. _{C19H13N3O2 . _ _} Calculated, %: C 72.31; H 4.12; N 13.33.

Example 2. Study of acute toxicity and anti-inflammatory activity of the claimed compound (I).

All research work with laboratory animals was carried out in accordance with generally accepted ethical standards for the treatment of animals adopted by the European Convention for the Protection of Vertebrate Animals Used for Research and Other Scientific Purposes [European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes, Strasbourg 1986 ].

Acute toxicity was studied on 8 nonlinear white mice weighing 21-24 g, using the express method of V.B. Prozorovsky [Prozorovsky, V. B. Express method for determining the average effective dose and its error / V. B. Prozorovsky, M. P. Prozorovskaya, V. I. Demchenko // Pharmacology and toxicology. - 1978. - No. 4. - P. 497-502.]. The test compound ( I ) was administered once intraperitoneally in 2% starchy mucus at doses of 500, 631, 794 and 1000 mg/kg. Then, the animals were monitored for 14 days, recording the intensity of motor activity, reactions to tactile, sound and light stimuli, the frequency and depth of respiratory movements, the condition of the hair and skin, the color of visible mucous membranes, water and food consumption [Guide to preclinical studies drug research / Ed. A.N. Mironova, N.D. Bunyatyan, A.N. Vasilyeva, O.L. Verstakova, M.V. Zhuravleva, V.K. Lepakhina, N.V. Korobova, V.A. Merkulova, S.N. Orekhova, I.V. Sakaeva, D.B. Utesheva, A.N. Yavorsky. M.: Grif i K, 2012. Part 1. 944 p.].

It was established that intraperitoneal administration of tested doses of the claimed compound ( I ) to animals did not cause clinical signs of its toxic effect, and also did not lead to the death of animals, therefore, LD ₅₀ for the claimed compound ( I ) is more than 1000 mg/kg. According to the classification of toxicity of substances, the claimed compound ( I ) belongs to toxicity class 5 and is a practically non-toxic compound [Sidorov, K. K. Pharmacology of new industrial chemicals. - 1973. - No. 13. - P.47 - 51.]. The data obtained allow us to conclude that the claimed compound ( I ) is superior to the comparison drugs - meloxicam and nimesulide. Thus, the minimum dose of meloxicam that can lead to the development of toxic effects when administered intraperitoneally is 1.25 mg/kg [Safety Data Sheet acc. to OSHA HCS [Electronic resource]. - Access mode: https://cdn.caymanchem.com/cdn/msds/14906m.pdf], and the average lethal dose of nimesulide when administered intraperitoneally is 163 mg/kg [Safety data sheet in accordance with GOST 30333-2007 [Electronic resource] . - Access mode: https://assets.lgcstandards.com/sysmaster%2Fpdfs%2Fh7c%2Fh14%2F10141283942430%2FSDS_MM0378.00_ST-WB-MSDS-2921057-1-1-1.PDF ].

The claimed compound ( I ) was tested for anti-inflammatory activity. The experiment was carried out on male rats weighing 220-280 g. An acute inflammatory reaction was caused by subplantar injection of 0.1 ml of 1% carrageenan solution into the hind paw of the animal [Guide to conducting preclinical studies of drugs / Ed. A.N. Mironova, N.D. Bunyatyan, A.N. Vasilyeva, O.L. Verstakova, M.V. Zhuravleva, V.K. Lepakhina, N.V. Korobova, V.A. Merkulova, S.N. Orekhova, I.V. Sakaeva, D.B. Utesheva, A.N. Yavorsky. M.: Grif i K, 2012. Part 1. 944 p.]. Foot volume in rats was measured oncometrically before administration of carrageenan (control), and then 3 and 5 hours after its administration. The test compound ( I ) was administered intraperitoneally as a suspension in 2% starch mucus at a dose of 50 mg/kg 30 min before the administration of phlogogen. The control group of animals was injected with an equivolume amount of 2% starch mucus. Nimesulide and meloxicam were used as comparison drugs in doses of 50 mg/kg each, administered intraperitoneally in 2% starch mucus. The anti-inflammatory effect was assessed by the indicator of inhibition of edema, which was expressed as a percentage of the control group. The research results are presented in Table 1.

In the course of the studies, it was revealed that the test substance ( I ) reduced the severity of carrageenan swelling of the paws of rats 3 and 5 hours after the induction of the inflammatory reaction. The severity of inhibition of the inflammatory reaction after 3 and 5 hours by the test compound ( I ) did not differ from that of the reference drug meloxicam, but was slightly inferior to the second reference drug - nimesulide.

Table 1
Anti-inflammatory activity of the compound ( I ) Test substance dose, mg/kg number of animals % inhibition of foot edema in 3 hours in 5 hours 3-methyl-5-oxo- N -phenyl- 5H -indeno[1,2- c ]pyridazine-4-carboxamide 50 6 67.22 ± 3.08* 73.01 ± 3.86* meloxicam 50 8 62.53 ± 5.95* 71.19 ± 8.08* nimesulide 50 6 87.25 ± 4.36* 93.67 ± 2.06* Note: * - p <0.05 relative to control;

Thus, the claimed compound 3-methyl-5-oxo- N -phenyl- 5H -indeno[1,2- c ]pyridazine-4-carboxamide ( I ), with low toxicity compared to comparison drugs, exhibits high anti-inflammatory activity.

The study was carried out with the financial support of the Perm Scientific and Educational Center “Rational Subsoil Use”, 2022.

Claims (4)

Use of 3-methyl-5-oxo- N -phenyl- 5H- indeno[1,2- c ]pyridazine-4-carboxamide of formula ( I ) I as a drug with anti-inflammatory activity.