RU2540464C2 - Water-soluble salamide derivative possessing tranquilising, nootropic and analgesic activity - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention can be used in chemical-pharmaceutical industry for preparing effective tranquilising salamide preparations and analgesics. What is presented is using salicyloyl morpholine sodium salt of formula (I)

as a tranquilising, nootropic and analgesic agent possessing low gastric toxicity.

EFFECT: implementing the declared application with the therapeutic index of salicyloyl morpholine of formula (I) is 3 times higher than that of mexidole, and 9,5 times higher than that of aspirin.

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Description

The invention relates to new water-soluble derivatives of salicylic acid amide (SC) and its use as a tranquilizing, nootropic and analgesic drug.

Salicylmorpholide (N-salicyloylmorpholine, CAS 3202-84-4) has an anti-inflammatory effect (US 3265575), but is not used in medical practice. In medical practice, among the used salicylamides, distant analogues of the claimed compound are known, such as salicylamide (CAS 65-45-2), which is prescribed orally as an analgesic and antipyretic, oxafenamide (CAS 526-18-1) - enhances the formation and secretion of bile, It has an antispasmodic effect and relieves or reduces biliary tract spasms, tiaprid and sultoprid, which have an antidopaminergic effect, are used externally as painkillers and anti-inflammatory drugs, sodium salt silic acid (enterzalil, diuretin, sodium salicylate, CAS 54-21-7) has an analgesic, antipyretic and anti-inflammatory effect [Mashkovsky MD Medicines: a manual for doctors. - T.1. - M .: New wave. - 2002. - S. 74, 167, 318, 504]. Acetylsalicylic acid (ASA) or Aspirin is the oldest non-steroidal anti-inflammatory drug with anti-inflammatory, analgesic, antiplatelet and antipyretic effects, and it is widely used for febrile conditions, headache, migraine, neuralgia [Kolosova O.A. Acetylsalicylic acid in the treatment of migraine // Journal. neur. and a psychiatrist. - 1998. - No. 4. - S. 44-46]. The lethal dose of aspirin, causing the death of 50% of animals (LD ₅₀ ), is 245.7 mg / kg [RU 2382763 C2]. Side effects of salicylates include nausea, gastralgia, allergic reactions, impaired liver function, Reye syndrome, etc. [Mashkovsky MD Medicines: a manual for doctors. - T.1. - M .: New wave. - 2002. - P.165].

Along with the listed therapeutic effects, which are manifested in almost all physiological systems of the body, salicylic acid and its derivatives also have a neurotropic effect. So, in the studies, the peculiarities of the neurotropic effect of cobalt salicylate and zinc salicylate on neurons of the grape snail were revealed [Korenyuk II. The effect of salicylic acid and its salts on the electrical activity of neurons of the grape snail / II Korenyuk, D. Khusainov, V. F. Shulgin // Neurophysiology - 2005. - T.37, No. 2 - S.142-150] .

However, in medical practice there are no SC drugs that would have a tranquilizing and nootropic effect.

The purpose of the invention is to obtain a water-soluble highly effective derivative of SC amide, combining the main effects inherent in tranquilizing agents, with a nootropic and analgesic effect, and characterized by low toxicity and acting in small doses.

The invention consists in the synthesis of the sodium salt of N-salicyloylmorpholine of the formula

and its use as a tranquilizing, nootropic and analgesic.

In studies of the tranquilizing and nootropic effects of the compound, doses of 10 and 50 mg / kg diluted in an isotonic solution of 0.9% NaCl at room temperature were used. The substance was administered once intraperitoneally 60 minutes before the start of the experiment. The results of the studies were statistically processed, the significance of the differences was evaluated using Student's criterion with Bonferroni correction.

Example 1. N-salicyloylmorpholine. In a three-necked reactor equipped with a stirrer and two dropping funnels, 57.9 mmol of morpholine are dissolved in 20 ml of 6 N. caustic soda solution. Within 30 minutes at a temperature of 5-10 ° C, 57.9 mmol of freshly prepared salicyloyl chloride and 6 N are simultaneously added. NaOH (to maintain pH 9.0 for universal indicator paper). After one hour exposure at a temperature of 5-10 ° C, the reaction mixture was extracted with ether (2 × 30 ml) and the aqueous layer was acidified with 1N HCl to pH 2.0 on universal indicator paper. Then it is cooled to 4 ° C, the precipitate is filtered off, washed with 60 ml of ether and dried at room temperature. The resulting technical N-salicyloylmorpholine was recrystallized from ethyl acetate. Get a white amorphous substance (yield 72%) with so pl. 165-168 ° C.

N-salicyloylmorpholine. ¹ H NMR spectrum (DMSO-d ₆ ), δ, ppm, J (Hz): 3.193-3.232 (4H, m, NCH ₂ , morpholyl), 3.328-3.510 (4H, m, OCH ₂ , morpholyl) 6.756-7.188 (4H, m, aromatic H); 9.784 (1H, s, Ph-OH).

Example 2. Sodium salt of N-salicyloylmorpholine. 4.6 g (100 mmol) of sodium ethylate, 35 g of toluene and 20.7 g (100 mmol) of N-salicyloylmorpholine are mixed and heated at 100 ° C. for 15 minutes with stirring. After cooling, the product is separated by filtration, washed with a small amount of benzene and dried. Get a white crystalline substance (22.2 g, 97%) with a melting point above 220 ° C. Sodium salt of N-salicyloylmorpholine. The ¹ H NMR spectrum (DMSO-d ₆ ) lacks a phenolic hydroxyl signal (1H, Ph-OH).

Example 3. The study of spontaneous-motor, research activity and emotional status in the test "open field".

This method allows you to study spontaneous locomotor activity, orientation and research behavior and the level of emotional response of animals. Within three minutes of observing the animal, the following indicators were recorded: the number of crossed squares (horizontal activity), the number of getting up on the hind legs and peeping in the holes (approximate research activity), the number of exits in the central zone, the number of grooming acts and the number of fecal boluses (emotional factor) (table 1).

Example 4. The method for determining the anxiolytic effect "plus" - a maze.

The method allows to evaluate the anxiolytic (tranquilizing, antiphobic) effect of psychotropic drugs. The labyrinth is raised above the floor to a height of 70 cm and represents a square platform (10 × 10 cm) with four cross-shaped sleeves 50 cm long and 10 cm wide. Two sleeves have opaque fences 40 cm high, two sleeves are open. The rat was placed on the central platform of the maze with its tail to the experimenter. Within 2 minutes of observation, the number of visits to open arms and the time spent in them were recorded. The increase in the number of exits into open arms and the time spent in them compared with those in the control group was evaluated as a manifestation of the anxiolytic (antiphobic) action of the substance (table 2).

Example 5. A method for studying nootropic activity is the development of a conditional passive avoidance reaction (passive avoidance reaction).

The study of the nootropic activity of compounds was carried out using a modification of this method, specially developed for rats (Bures, Buresova, 1979). The development of a conditioned reaction to avoid a darkened compartment was carried out in an experimental chamber, which consisted of two adjacent compartments - a large illuminated (60 × 40 cm) and a small darkened (15 × 15 cm) equipped with an electrode floor. The compartments were interconnected using a quadrangular hole (8 × 8 cm). The rat was placed in the light compartment, with its tail toward the dark compartment, and the latent period of the first entry of the animal into the dark (dangerous) compartment was recorded for 3 minutes. Rats that preferred a bright compartment were excluded from further experiments. After three minutes in the dark compartment, the animal received a single electric pain stimulation (40 V) through the electrode floor, consisting of three pulses of 1 second duration, followed by an interval of 0.5 seconds. A rat was considered trained if, within 30 seconds after the training session, it did not enter the dark compartment of the experimental chamber.

A test for reproducing a commemorative footprint was carried out 24 hours after training. The animal was placed in the bright compartment of the chamber and the same parameters were recorded for 3 minutes as during training. The nootropic effect of the compound was manifested when reproducing the conditional passive avoidance reaction (passive avoidance reaction) and was expressed in an increase of 88.6% in the latent period of the first entry of the animal into the dark compartment and a 55% decrease in the time spent in it compared to animals in the control group (table 3).

Example 6. A method for studying analgesic activity. The method allows to evaluate the analgesic activity of compounds by changing the threshold of vocalization with a gradual increase in AC voltage before the occurrence of electric pain irritation and vocalization. After placing the animal in the installation, electric current pulses were supplied with a gradual increase in voltage (rise rate of 5 V / s, pulse duration - 0.2 seconds). The tolerance threshold for electric pain stimulation was recorded when vocalization appeared in the animal.

The inventive compound - sodium salt of N-salicyloylmorpholine - at a dose of 50 mg / kg significantly compared with the control, increased vertical and horizontal activity (by 106.1% and 90.8%, respectively) in the test "open field", which can be regarded as an increase in research activity. This compound at a dose of 50 mg / kg significantly compared with the control, increased the number of exits and the time spent in the open arms of the raised “+ maze” (by 128.8% and 47.2%, respectively), which is regarded as a manifestation of pronounced tranquilizing activity . This compound also significantly increased the rat vocalization threshold (by 88.6%), which indicates the presence of its high analgesic activity. Of particular interest is the combination of tranquilizing and analgesic activity, which have a number of similar mechanisms for their implementation (table 4).

Example 7. Determination of acute toxicity. Acute daily toxicity _(LD50) of the sodium salt of N-salitsiloilmorfolina determined on white mice weighing 25-30 g were examined substance in doses of 1000 to 3200 mg / kg. For testing, the substance substances were dissolved in an isotonic solution of 0.9% NaCl at a temperature of 20 ° C "extempora". After dissolution, the test solution was administered to mice intragastrically or intraperitoneally. Along with experimental animals that received the test substance, control animals (intact group) were kept under similar conditions. The control animals, in equivalent volumes, were injected intragastrically and intraperitoneally with a solvent, an isotonic solution of 0.9% NaCl.

After administration of the test substances, the mice were intragastrically monitored for 15 days. Moreover, observations on the first day were carried out continuously throughout the entire daylight hours, and periodically on the next day. The clinic of poisoning was noted, its duration and the number of deaths. Pay attention to behavior, nutritional activity, dynamics of body weight. When administered orally, the dead animals were opened, a macroscopic review of the internal organs was performed. At autopsy, the state of parenchymal organs was examined macroscopically (their coloration, location, size were noted). Since derivatives of salicylic acid can have an ulcerogenic effect, a macroscopic examination of organs drew attention to the state of the gastric mucosa. Studies of acute toxicity of the claimed water-soluble derivative of amide SC showed that, according to the classification of the hazard (harmfulness) of substances, the sodium salt of N-salicyloylmorpholine when administered intragastrically to mice belongs to the class of low toxic compounds [Sanotsky I.V. The harmfulness criterion in hygiene and toxicology when assessing the hazard of chemical compounds / I.V. Sanotsky, I.P. Ulanova. - M .: Medicine, 1975 - 328 p.] (Table 5).

Domestic drug "Meksidol" (emoxypine) exhibits a nootropic, neuroprotective, anxiolytic, antihypoxic, antistress, anti-alcohol, antiparkinsonian and anticonvulsant activity and others. However, the lethal dose mexidol causing the death of 50% of animals (LD ₅₀₎ of rats 820 ( 625-1025) mg / kg and for mice 475 (365-617) mg / kg, and when administered orally, more than 3000 mg / kg in rats and 2010 (1608-2513) mg / kg in mice. The therapeutic index, calculated by the ratio of LD ₅₀ / ED ₅₀ is 16.4 [T. Voronina Domestic drug of the new generation Mexidol: main effects, mechanism of action, application. - M .: 2004. - 248 p.]. Mexidol does not have an analgesic effect.

From a comparison of the doses in which the N-salicyloylmorpholine sodium salt exhibits tranquilizing and nootropic properties in combination with the analgesic effect (50 mg / kg) with LD ₅₀ (2341.124 mg / kg orally and 2550.400 mg / kg intraperitoneally), we can conclude that the drug has a wide range of therapeutic effects and significantly lower toxicity. Thus, the toxicity (when administered intraperitoneally) of mexidol is 5 times higher than that of the claimed compound, and the therapeutic index of the claimed compound is almost 3 times higher than the index of mexidol and almost 9.5 times higher than that of aspirin (table 5).

In addition, the salt water-soluble form of the compound (sodium salt of N-salicyloylmorpholine), unlike Aspirin, allows its use in the form of injections, avoiding the effects on the gastrointestinal tract.

Table 1 The effect of sodium salt of N-salicyloylmorpholine on the behavior of rats in the "open field", (M ± m) Compound Dose mg / kg Vertical motor activity (the number of stands on the hind legs) Horizontal motor activity (number of crossed squares) The number of peeping holes The number of grooming acts The number of exits to the central zone Number of boluses The control 0 7.0 ± 0.83 25.38 ± 4.67 2.25 ± 0.42 7.38 ± 0.7 0.38 ± 0.17 2.75 ± 0.72 Sodium salt of N-salicyloylmorpholine 10 8.75 ± 1.74 35.25 ± 6.09 2.25 ± 0.42 5.38 ± 0.83 0.38 ± 0.72 1.75 ± 0.52 fifty 14.43 ± 1.05 * 48.43 ± 6.83 * 2.86 ± 0.59 3.43 ± 0.9 0.57 ± 0.34 1.43 ± 0.63 * - the data are statistically reliable, Student's test with Bonferroni correction, p <0.05

table 2 The anxiolytic effect in the test "plus" -labyrinth of the sodium salt of N-salicyloylmorpholine Compound Dose mg / kg Exit to open sleeves Stay in open sleeves The control 0 1.25 ± 0.49 21.25 ± 8.88 Sodium salt of N-salicyloylmorpholine 10 1.14 ± 0.31 16.00 ± 5.49 fifty 2.86 ± 0.24 * 31.29 ± 5.39 * * - the data are statistically reliable, Student's test with Bonferroni correction, p <0.05

Table 3 Anti-amnestic effect in the test of the conditional reaction of passive avoidance of the sodium salt of N-salicyloylmorpholine Compound Dose mg / kg Latent period of entry into the dark compartment Stay in the dark compartment The control 0 19.13 ± 3.2 160.88 ± 3.2 Sodium salt of N-salicyloylmorpholine 10 108.0 ± 31.44 * 72.0 ± 31.44 * fifty 35.0 ± 15.2 145.0 ± 15.2 * - the data are statistically reliable, Student's test with Bonferroni correction, p <0.05

Table 4 The effect of the sodium salt of N-salicyloylmorpholine on the vocalization threshold, (M ± m) Compound Dose mg / kg Vocalization threshold, volt The control 0 32.57 ± 2.17 Sodium salt of N-salicyloylmorpholine 10 28.57 ± 3.01 fifty 61.43 ± 5.88 * * - the data are statistically reliable, Student's test with Bonferroni correction, p <0.05

Table 5 Comparative characteristics of the effectiveness of the sodium salt of N-salicyloylmorpholine / mexidol and the sodium salt of N-salicyloylmorpholine / acetylsalicylic acid (aspirin) LD ₅₀ , mg / kg

Therapeutic index $$\frac{L{D}_{\mathrm{fifty}}}{E{D}_{\mathrm{fifty}}}$$

Sodium salt of N-salicyloylmorpholine (intraperitoneally) 2550,400 * (2294,611 ÷ 2834,701) 51.00 Mexidol (intraperitoneally) 475 (365-617) 16,4 Sodium salt of N-salicyloylmorpholine (intragastrically) 2341.124 * (3676.349 ÷ 1490.844) 46.82 Acetylsalicylic acid (aspirin) (intragastric) 245.7 * (287.4 ÷ 210.1) 4.91 Attitude $$\frac{\mathrm{from}\mathrm{about}lb}{me\mathrm{to}\mathrm{from}\mathrm{and}d\mathrm{about}l}$$

5.37 $$\frac{5,37}{\mathrm{one}}$$

3.11 Attitude $$\frac{\mathrm{from}\mathrm{about}lb}{\mathrm{but}\mathrm{from}P\mathrm{and}R\mathrm{and}n}$$

9.53 $$\frac{9,53}{\mathrm{one}}$$

9.54 * - data are statistically significant, p <0.05

Claims (1)

The use of the sodium salt of salicyloylmorpholine of the formula

as a tranquilizing, nootropic and analgesic.