RU2605265C1 - Analgesic and anti-inflammatory agent based on 5-butyl-6-hydroxy-2-methyl pyrimidine-4(3h)-one - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine and pharmacy and represents analgesic and anti-inflammatory agent, containing 5-butyl-6-hydroxy-2-methylpyrimidine-4(3H)-one as active substance.

EFFECT: agent is low-toxic, exhibits considerably higher anti-inflammatory and analgesic activity than preparations of comparison, widely used in medicine.

1 cl, 4 ex

Description

The invention relates to medicine and pharmacy and relates to a new analgesic and anti-inflammatory agent based on a dihydroxypyrimidine derivative, namely 5-butyl-6-hydroxy-2-methylpyrimidin-4 (3H) -one (C ₉ H ₁₄ N ₂ O ₂ ) of formula I :

2,5-disubstituted 6-hydroxypyrimidin-4 (3H) -ones, their structure, properties and methods of preparation are widely known [Livingston P. Ferris II, Anthony R. Ronzio // Contribution from the Chemistry Department of the University of Colorado. 1940. P. 606-607; Kaushik S. / Patel, Kishor N. Raval, Shivani P. Patel, Aswin G. Patel, Snehal V. Patel // AvailableOnlinethrough. 2012. P. 170-182]. However, nothing is known about their biological activity, and, in particular, about the analgesic and anti-inflammatory properties.

The anti-inflammatory activity of such compounds belonging to the class of dihydroxypyrimidines as oxymethyluracil is known [D.N. Lazareva, V.A. Davydova // Experimental and Clinical Pharmacology. 2005. S. 53-55], as well as a number of 1,3-alkyl-5-hydroxy-6-methylpyrimidin-2,4 (1H, 3H) -dione [Patent 2051150 RF, C07D 413/06, C07D 211 / 22, A61K 31/505. Pyrimidine acyclonucleosides exhibiting immunotropic and anti-inflammatory activity].

The closest in structure and pharmacological properties to the claimed agent is oxymethyluracil of formula II (prototype)

possessing anti-inflammatory activity against histamine (64.8%), serotonin (75.0%), bradycardin (65.7%), carrageenan (50.0%) and trypsin (97.8%) edema.

The disadvantages of this compound include a fairly high toxicity, as well as the absence of any data on its analgesic activity.

The objective of the invention is the expansion of the arsenal of analgesic and anti-inflammatory drugs, the search for a new agent based on dihydroxypyrimidine, which has low toxicity, has analgesic and anti-inflammatory activity, exceeding in their indications the indicated activities used separately in medical practice.

This problem is achieved by creating an analgesic and anti-inflammatory agent based on 5-butyl-6-hydroxy-2-methylpyrimidin-4 (3H) -one.

The invention is illustrated by examples of synthesis and study of pharmacological properties.

Example 1. Method for the synthesis of 5-butyl-6-hydroxy-2-methylpyrimidin-4 (3H) -one.

In a round-bottomed three-necked 250 ml flask equipped with a reflux condenser, a dropping funnel and a thermometer, 9.86 g (0.17 mol) of acetamidine (acetamidine, due to its high price, can be obtained from the acetimidoether and alcoholic ammonia solution [E .V. Kuvaeva, E.V. Fedorova, V.V. Zaitsev, I.P. Yakovlev, V.I. Zakharov, T.L. Semakova. - Journal of Organic Chemistry, St. Petersburg, 2011.V. 48. P. 221 -225] and isolated from acetamidine hydrochloride by reaction with an aqueous solution of potash in absolute ether), and 50 ml of benzene are added (GOST 5955 -75) absolute. The stirrer is turned on and 43.34 g (0.22 mol) of freshly prepared [Organic Syntheses, Coll. Vol. 4, p. 261 (1963); Vol. 34, p. 26 (1954)] 2-butyl malonoyl dichloride. The reaction mass is heated at a temperature of 50 ° C for 60 minutes. As the reaction proceeds, a precipitation of the target pyrimidine is observed. The resulting 5-butyl-6-hydroxy-2-methylpyrimidin-4 (3H) -one is purified by reprecipitation. The precipitate is dissolved in 100 ml of a 15% sodium hydroxide solution, and then acidified with 26% hydrochloric acid to pH 4. The precipitate is filtered off and dried in an oven at 70 ° C. A fine cream-colored powder is obtained. The product yield was 87% based on acetamidine.

The chromatographic homogeneity of the target product was confirmed by chromatography of a solution of it in DMSO in the system acetone: glacial acetic acid 4: 1 (R _f = 0.61). The composition of the synthesized compound is confirmed by elemental analysis. Gross formula: C ₉ H ₁₄ O ₂ N ₂ . Found%: C - 61.04; H - 7.09%; N - 14.17%. Calculated%: C - 59.35%; H - 7.69%; N - 15.38%.

The structure of the synthesized substance was proved by physicochemical methods for the identification of organic compounds: ¹ H and ¹³ C NMR, UV and IR spectroscopy.

In the IR spectra of the substance (KBr tablets), the most characteristic are absorption bands in the region of 3500-3080 cm ^-1 , characterizing the presence of OH and NH groups, 1670-1680 cm ^-1 , characteristic of the C = O group, and 1550-1558 cm ^{- 1} , characteristic for the bond C = N.

The UV spectrum of 5-butyl-6-hydroxy-2-methylpyrimidin-4 (3H) -one in 99.6% acetonitrile has 1 absorption maximum at 262 nm.

The ¹ H NMR spectrum of the obtained compound in DMSO-d ^{6 is} characterized by the presence of proton signals (0.83 3H, s; 1.20 4H, m; 2.11 2H, t) butyl substituent, (2.66 3H, s) methyl protons groups, as well as peaks in the region of (8.96 1H, s; 9.11 1H, s) protons of NH and OH groups, respectively.

The ¹³ C NMR spectrum of this compound is characterized by the signals of the nuclei of the carbon atoms of the butyl substituent (δ 14.06; 22.38; 28.80; 48.91 ppm), the signal of the carbon atom of the methyl group (δ 29.93 ppm) , as well as carbon atoms C ⁵ (δ 99.04 ppm), C ² (δ 155.83 ppm), C ⁴ (δ 164.42 ppm) and C ⁶ (δ 175 , 04 ppm).

The structure of the obtained substance was determined using mass spectrometry. The calculated molecular weight completely coincided with the experimentally obtained (M ⁺ = 244).

Example 2. Determination of acute toxicity.

The acute toxicity of the synthesized compound was determined on non-linear white male mice with a body weight of 18-20 g. Animals were divided into groups of equal size and body weight, 10 animals each. Suspensions of the compound in water stabilized with tween 80 were administered once intraperitoneally at a dose range of 50 mg / kg to 2000 mg / kg. The survival of animals was determined by observing after 24 hours and after 48 hours from the moment of administration of the test compound. Observation of the animals was carried out for 72 hours. The development of the main symptoms and the time of death of the animals were recorded.

The calculation of the lethal dose (LD ₅₀ ) was carried out using the express method V.B. Prozorovsky [Prozorovsky VB, Prozorovskaya M.P., Demchenko V.I. // Pharmacology and toxicology. - 1978. - No. 4. - S. 497-502] and probit analysis according to the Miller-Taint method [M.L. Belenky. Elements of a quantitative assessment of the pharmacological effect. (2nd edition, revised and supplemented). State Publishing House of Medical Literature. Leningrad. 1963. p. 412]. The toxicity of the claimed substance is 1873 mg / kg. According to the classification of toxicity of drugs, compound I belongs to the class of low toxic substances [Izmerov I.F., Sanotsky I.V., Sidorov K.K. Toxicometry parameters of industrial poisons. M .: Medicine, 1977. S. 196-197]. It is more than 20 times less toxic than diclofenac sodium - a comparison drug for anti-inflammatory activity.

Example 3. Determination of analgesic activity.

For experimental evaluation of analgesic activity, the model of generation of acetic acid “writhing” in male mice was used. Seizures in animals were caused by intraperitoneal administration of a 3% solution of acetic acid. The experiment was carried out on white non-linear male mice weighing 15-22 g. For the experiment, 3 groups of animals of 7 animals each were used. A suspension of 5-butyl-6-hydroxy-2-methylpyrimidin-4 (3H) -one in a mixture of DMSO with water (1: 5) (tween stabilizer 80) was administered once, intraperitoneally at doses of 1/10 LD ₅₀ 40 minutes before intraperitoneal the introduction of a 3% solution of acetic acid. The animals of the control group were injected with a mixture of DMSO: water (1: 5). Metamezole sodium at a dose of 100 mg / ml was used as a comparison drug. The time of onset of seizures and their number within 20 minutes were recorded [D. Kharkevich Pharmacology Textbook. - 10th ed. - M.: GEOTAR-Media, 2009. - S. 628-640]. The analgesic activity of the claimed substance is 48%, which is slightly lower than the metamezole sodium comparison drug, however, the test substance is more than 20 times less toxic.

Example 4. Determination of anti-inflammatory activity.

Anti-inflammatory activity was investigated on a screening model - formalin paw edema in rats.

In the experiment used white outbred male rats weighing 200-240 g in an amount of 40 pieces. Of these, 4 groups of 10 animals each were formed. The control group was administered only the phlogogenic agent. Animals of the experimental groups received the study drug in doses of 1/10 LD ₅₀ , 1/50 LD ₅₀ and the comparison drug, which was administered intraperitoneally 1 hour before the edema was created.

An aqueous 2% formalin solution was administered subplanarly in an amount of 0.1 ml to each animal.

The studied native solution of (5-butyl-6-hydroxy-2-methylpyrimidin-4 (3H) -one) was administered to rats intraperitoneally at doses of 190 and 40 mg / kg. Comparison drug diclofenac was administered at a dose of 25 mg / kg.

Experimental procedures were performed on each animal in the following sequence:

- intraperitoneal administration of the test sample: in an acute experiment, once 1 hour before the introduction of the phlogogenic substance;

- measurement of the volume of the foot of the right hind paw of the animal (source data);

- subplant implantation of formalin (0.1 ml of a 2% solution) into the animal’s right hind paw one hour after the last administration of the test sample;

- measurement of the foot volume of the right hind paw of the rat at certain intervals after administration of the phlogogenic agent (1 and 24 hours).

The paw volume was measured using a glass oncometer according to the volume of displaced water after the paws were immersed in a special tank.

The results obtained from each animal were averaged within the group and the percentage of inhibition of inflammation in the experimental group relative to the control was calculated.

A single intraperitoneal preventive administration of 5-butyl-6-hydroxy-2-methylpyrimidin-4 (3H) -one in a dose of 1/10 LD ₅₀ helps to inhibit the development of the inflammatory reaction after 1 hour by 11.0% (1/10 LD ₅₀ ) and 15.1% (1/50 LD ₅₀ ), which is inferior to the reference drug with an indication of 25%, after a day by 97.4% (1/10 LD ₅₀ ) and 96.1% (1/50 LD ₅₀ ), which is significantly exceeds the indications of sodium diclofenac widely used in practice (60.8%) with significantly lower toxicity of the claimed drug. It has also been reliably shown that a dose reduction does not lead to a decrease in the biological activity of the test compound, and in the first hours it even slightly increases the effectiveness of the claimed substance.

Thus, the claimed tool is low toxic, exhibits significantly greater anti-inflammatory activity than the reference drug, widely used in medicine. In addition, the claimed substance has a noticeable analgesic effect with significantly lower toxicity than the widely used comparison drug in practice.

Claims (1)

Analgesic and anti-inflammatory, including 5-butyl-6-hydroxy-2-methylpyrimidin-4 (3H) -one as an active substance.