RU2758333C1 - Quinazolin-4(3h)-one dimethoxybenzylsulfo derivative with analgesic, antiparkinsonian, anxiolytic, psychostimulating activity - Google Patents

Abstract

FIELD: organic chemistry.

SUBSTANCE: invention relates to the field of organic chemistry, namely to a heterocyclic compound 2-(3,4-dimethoxybenzyl-6-sulfo)quinazolin-4(3H)-one, corresponding to formula (1). The invention also relates to a method for its preparation.

EFFECT: obtained a new heterocyclic compound with analgesic, antiparkinsonian, tranquilizing, psychostimulating activity.

2 cl, 11 tbl

Description

The present invention relates to a new dimethoxybenzylsulfo-derivative of quinazolin-4 (3H) -one, characterized by high analgesic, antiparkinsonian, anxiolytic and psychostimulating activity in combination with low toxicity, and therefore can find application in the pharmaceutical industry and medicine. It concerns the use of a benzyl thio derivative of quinazolin-4 (3H) -one for the production of drugs that can be used as pain relievers, antiparkinsonian, psychostimulating drugs, and daytime tranquilizers. The compound can be used for the preparation of drugs for the treatment of pain syndrome; parkinsonism; with neuroses, neurosis-like states, psychosomatic disorders, accompanied by emotional stress disorders and asthenia. The invention expands the number of drugs with psychostimulating activity.

Prior art

The results of a scientific search for substances that affect the central nervous system, among purposefully synthesized biologically active compounds, are in demand and are determined by the urgency of the problem of creating new domestic original drugs. The creation of new analgesic drugs - safe and highly effective - is an urgent task for pharmacology and pharmacy, which can be solved by the synthesis and research of new biologically active compounds [1-4]. Derivatives of 4- (3) quinazolinone are known that affect the central nervous system, but have significant toxicity [5].

In the search for neurotropic substances, the emphasis is placed on the targeted synthesis of compounds exhibiting agonistic activity at dopamine receptors. The increase in the number of people of older age groups in the structure of society is accompanied by an increase in the incidence of Parkinson's disease, which has become a social problem. In the pharmacotherapy of Parkinson's disease in the early stages, it is recommended to use dopaminomimetic agents that can exhibit neuroprotective activity, protect neurons from negative effects, and restore damaged nerve cells. Neuroprotective activity has been established for a number of quinazolinone-4 derivatives [6,7]. Researchers are interested in hydroxy- and methoxyphenyl derivatives of quinazolinone-4, which may have dopaminergic activity.

The development of new drugs with anti-anxiety effects will make it possible to improve the drug treatment of anxiety disorders, which often accompany depressive disorders and other neuropsychiatric pathological conditions. Existing anxiolytics have a pronounced sedative effect, amnestic effect, cause adverse changes in the blood picture, etc., are not recommended for activities associated with the need for concentration. In this regard, compounds that combine 2 or more types of positive action in the spectrum of activity will be advantageous.

The closest in structure to the claimed compound are hydroxy and methoxy derivatives of 4- (3) quinazolinone, which have dopaminergic activity [8].

These data indicate that the search for new effective and safe analgesic, antiparkinsonian substances with anxiolytic properties among 3H-quinazolin-4-one derivatives is promising.

The present invention relates to a new sulfo derivative of quinazolin-4 (3H) -one, which has analgesic, antiparkinsonian, psychostimulating properties in combination with tranquilizing properties and practically no toxicity.

EFFECT: obtained a new sulfo derivative of 4- (3H) quinazolinone, which has an analgesic effect, which in tests with formalin, "vinegar cramps" is superior to the comparison drugs metamizole sodium and lidocaine. The compound exhibits antiparkinsonian properties, surpassing the anticataleptogenic effect of bromocriptine, levodopa in the haloperidol catalepsy test. The resulting compound also has a tranquilizing effect, which is superior to the reference drug diazepam in the "elevated cruciform labyrinth", "open field" tests, multiparametric testing. The newly obtained compound has a psychostimulating effect, superior in the "lateral position" test to the prototype caffeine sodium benzoate. Compound 1 was obtained by a new method.

The claimed object is 2- (3,4-dimethoxybenzyl-6-sulfo) quinazolin-4 (3H) -one of formula (1), which has analgesic, antiparkisonic, anxiolytic and psychostimulating activity.

The method of obtaining 2- (3,4-dimethoxybenzyl-6-sulfono) -quinazolin-4 (3H) -one (1) is based on the interaction of 2- (3,4-dimethoxybenzyl) - (3H) -quinazolin-4-one [ 9] with concentrated sulfuric acid at room temperature, followed by the isolation of the reaction product by diluting the reaction mixture with water.

SCHEME OF OBTAINING

The new compound, the thio derivative of 4- (3H) -quinazolinone, exhibits analgesic, antiparkinsonian, anxiolytic and psychostimulating effects.

EXAMPLE OF SYNTHESIS

At room temperature, with stirring, dissolve 3.0 g (0.01 mol) of 2- (3,4-dimethoxybenzyl) quinazolin-4 (3H) -one in 15 ml of concentrated sulfuric acid. The homogeneous reaction mixture is incubated for 10-12 hours at room temperature and poured into 150 g of a mixture of ice water. The formed colorless precipitate is filtered off, washed thoroughly on the filter with water and isopropyl alcohol. The air-dried product is then used without further purification. 80% yield. Mp> 300 ° C.¹H NMR (600 MHz, DMSO-d₆ , δ, ppm): 3.55 (s, 6H, 2OCH₃), 4.70 (s, 2H, CH₂), 6.94 (s, 1H, ar.), 7.38 (s, 1H, ar.), 7.20-7.55 (d. + T, 2H, ar.), 7.83 (t, 1H, ar.), 8.25 (d . 2H, ar.).

Assessment of pharmacological activity

The pharmacological activity of the compound was evaluated in four directions. The analgesic, antiparkinsonian, anxiolytic, psychostimulating effects of compound 1 have been studied.

For the research, white sexually mature male rats (5-11 per group) and females (10-12 per group) of the Wistar line weighing 200-250 g were used, which were kept with free access to water and food under standard vivarium conditions. Care, feeding, work with animals was carried out in accordance with international recommendations for the protection of vertebrate animals used for experiments or for other scientific purposes [10-12].

The control animals in each series of experiments for each method in assessing the studied activity were groups of rats - females, males, who were injected intraperitoneally with a solvent in a volume of 0.4 ml (1-2 drops of Tween-80 per 10 ml of water) or saline (control group for animals that received prototypes in similar modes - reference drugs (metamizole sodium, lidocaine, bromocriptine, diazepam). Groups of animals consisted of 5-12 individuals (experimental, control, reference drugs). All solutions of substances in all series of studies were injected intraperitoneally. C Taking into account the chronotropic activity of psychotropic drugs, experiments were carried out in the daytime (from 12-00 to 17-00) and in the evening (from 18-00 to 20-00) [13].

Statistical analysis of the research results was carried out using the software packages "Excel", "BIOSTAT", "Statistica 10". The normality of the distribution of the obtained data was determined using the Shapiro-Wilk test. Student's t-test was used with normal distribution of the obtained data. When the distribution of the research data was different from the normal, the Kruskal-Wallis test and the Wilcoxon-Mann-Whitney U-test were applied. A comparative analysis was carried out, during which the results of the experimental groups were compared with the data of the control animals (taken equal to 100%). Comparative analysis of differences in the action of substances in female animals was also carried out at the stages of the estrous cycle diestrus ½ and proestrus / estrus. Differences were taken statistically significant at p <0.05 for all types of analysis [14-16].

The analgesic activity of compound 1 was studied in formalin and vinegar cramps tests; antiparkinsonian action - on the model of haloperidol catalepsy; tranquilizing activity - in tests of an elevated cruciform labyrinth, "open field", multiparametric testing, psychostimulating activity - on the model of "lateral position" [11,12].

Analgesic activity

The influence of the claimed object on analgesic activity was carried out on models of nociceptive reactions caused by chemical stimuli (formalin test and "vinegar cramps"). The tests were used to assess the peripheral level of pain sensitivity [17].

The formalin test allows assessing the somatic pain induced by algogens and simulates the response to surgical skin incisions. The first (acute) stage develops immediately under the influence of the pain factor, manifests itself as an effect on the primary afferents of pain; the second (inflammatory) stage is delayed in time and is caused by an inflammatory response. Pain sensitivity was simulated by subplantar injection of 50 μl of an aqueous solution of 2% formalin into the dorsum of the hind right paw. Subsequently, nociceptive responses ("flinches": raising the paw, licking, biting the injection site) were recorded within 60 minutes of observation: 1) the first 10 minutes from the moment of formalin administration (acute phase); 2) from the 10th to the 60th minute of the pain reaction (inflammatory phase). The analgesic effect of the test compound was determined in total and separately for phases I and II of the pain response by reducing the number of responses in relation to the indicators of the control group of animals, taken as 100%.

The severity of peritovisceral pain was assessed under the conditions of the "Acetic cramps" model. Using this method of conventional visceral nociception, the peripheral analgesic activity of new substances was investigated using a chemical algogen to irritate the nocireceptors of the peritoneum, and specific motor responses - “cramps” - were evaluated. "Writhing" are characteristic painful behavioral responses that arise under the influence of chemical stimuli on the serous membranes, are manifested by contraction and relaxation of the abdominal muscles, stretching of the hind legs, back arching, similar to peritoneal pain. "Writhing" was induced by a single injection of 1% acetic acid solution (0.5 ml / 100 g of animal body weight; intraperitoneally). The analgesic effect was assessed by a decrease in the number of "writhing" within 15 minutes of observation after the administration of an acetic acid solution (in% to the indicators of the control group of animals, taken equal to 100%).

Substance 1 at a dose of 0.2 of the molecular weight in mg / kg (75.2 mg / kg), reference drugs in therapeutically effective doses: metamizole sodium - 55 mg / kg (Metamizolum natrium; analgin, solution for intravenous and intramuscular administration 500 mg / ml ampoule 2 ml; "Moskhimpharmaceuticals named after N. A. Semashko", Russia), lidocaine - 1 mg / kg (Lidocainum; ampoules 2% 2 ml; "Dalkhimpharm", Russia) was injected intraperitoneally once 40 minutes before introduction of chemical irritants. The dose of the investigated compound 1 and reference drugs were selected by the dose titration method and taking into account the literature data [18-19]. Males and females of the control groups were injected, respectively, with Tween-80 solubilizate and distilled water (0.4 ml intraperitoneally) in similar modes. Taking into account the differences in the activity of drugs depending on sex, phases of the estrous cycle, studies were carried out in male and female rats, including females in different phases of the estrous cycle (EC). Prior to testing, the phases of the estrous cycle (proestrus / estrus (PE) and diestrus ½ (D)) were determined using vaginal smears of female animals [13]. The experiments were carried out in the daytime.

When studying the analgesic activity by tests with formalin and "vinegar cramps", a high activity of compound 1 was noted (table 1, 2). Compared with the data of the group of control males, when using compound 1 in the formalin test, the sensation of pain was suppressed, more effective than analogs: more than 1.42 times metamizole sodium and 2.61 times - lidocaine. In the test "vinegar cramps" when using compound 1 in comparison with the data of males in the control group, the sensation of pain was limited more effectively than the analogue of lidocaine by 4.66 times.

When testing "vinegar writhing" against the background of the use of compound 1 in females, the frequency of pain responses was statistically significantly limited (Table 3). In this case, the effect of compound 1 was more pronounced in comparison with the effect of lidocaine.

When analyzing the analgesic activity of the studied substance 1 in female animals, taking into account the phase of the estrous cycle, a statistically significant decrease in the number of pain reactions was noted. When using compound 1 in females, the number of pain responses in general and throughout all stages of testing with formalin decreased, statistically significant in the stage of physiological rest - diestrus ½ (Table 4). At the same time, the decrease in the frequency of raising the paw, licking, and biting the injection site under the influence of compound 1 was more pronounced in comparison with the effects of the reference drug.

In the test "vinegar writhing" with the introduction of compound 1, the number of "writhings" was significantly limited in female rats in diestrus ½ (Table 5). At the same time, the decrease in the number of "vinegar writhing" was more pronounced in comparison with the action of lidocaine in female animals in the phase of the estrous cycle diestrus ½.

Consequently, compound 1 reduced the frequency of pain responses in formalin tests and "vinegar cramps" in female animals, exhibiting analgesic activity.

Conclusion: compound 1 has an analgesic activity that exceeds the effect of the reference drugs metamizole sodium, lidocaine.

Anti-cataleptic activity

The antiparkinsonian (anticataleptic) effect of compound 1 was studied in sexually mature female white rats (Wistar lines, weighing 200-250 g, 9-11 in groups) in the haloperidol catalepsy test (haloperidol at a dose of 1 mg / kg intraperitoneally) [13]. Compound 1 at a dose of 75.2 mg / kg (in suspension with Tween-80), reference preparations bromocriptine (at a dose of 0.125 mg / kg) (Bromocriptinum, 2.5 mg tablets, Gedeon Richter, Hungary), levodopa (in dose 10 mg / kg) (Syndopa, tablets 250 mg, Levodopum 250 mg + Carbidopum 25 mg, Sun Pharmaceutical Industries Ltd., India) were administered intraperitoneally simultaneously with haloperidol (Haloperidolum, tablets 1.5 mg, Biocom, Russia ). The groups of control females were injected with a neuroleptic in a similar regimen. The experiments were carried out during the daytime.

The dynamics of movement disorders in animals was determined against the background of the studied substances. Changes in the severity of haloperidol catalepsy - latent periods of time (in seconds, s) of verticalization (in the "lecturer's position") - the duration of the animals retaining an upright position on the support were assessed. The periods of time after which the animal removed one / second paw were established: 1st / 2nd results. The registration of the results was carried out after the introduction of the solubilizate of compound 1, a solution of bromocriptine comparison drugs, sindopa after 30, 60, 120, 180 minutes [11, 12]. The phases of the estrous cycle in female rats (proestrus / estrus, diestrus ½) were determined by vaginal smears.

When using a combination of compound 1 with a neuroleptic at the 120th and 180th minutes of the test, movement disorders were weakened, statistically significant at the 120th minute of testing (Tables 6, 7). The effect was more pronounced than with the introduction of comparison drugs.

Conclusion: compound 1 has anticataleptogenic activity, which exceeds the effect of bromocriptine and levodopa preparations in the studied doses.

Anxiolytic activity

Evaluation of the tranquilizing activity of compound 1 according to the elevated cruciform maze (PCL) test [20] was carried out in an experimental setup consisting of 4 arms, 50 × 10 cm in size, fastened at right angles to each other. Two opposite PCL sleeves are closed (fenced on both sides, dark) and the other two are open (light). The labyrinth is raised above the floor level to a height of 0.5 m. The height of the walls in the closed sleeves is 10 cm. The sleeves are open at the top. At the intersection (at right angles) of the arms, there is a platform with dimensions of 10 × 10 cm.During testing, the animals were placed on the central platform of the maze, and then behavioral parameters were visually recorded for 5 min: the total number of entries into the arms of the maze (open or closed) and the time finding them (c). The anxiolytic activity of the test compound was determined by the increased frequency of entry of the animal into the light (open) arms and an increase in the residence time in them, a decrease in the number of peepings and the time spent in the dark (closed) arms [11, 12].

The study was carried out on male white rats. Compound 1 at a dose of 37.6 mg / kg (0.1 of the molecular weight in mg / kg), the reference drug is prototype diazepam at doses of 0.1 and 0.5 mg / kg (Diazepamum, solution for intravenous and intramuscular administration 10 mg / 2 ml; ampoules 2 ml; "Relanium", "Polfa", Poland) were injected once intraperitoneally 40 minutes before the start of testing. Control males were injected intraperitoneally with physiological saline in similar modes.

With the introduction of compound 1, the time spent in the dark arms of the maze was limited, while the frequency of entry into them was reduced in comparison with the behavior parameters of the control animals of the K1 group. Against the background of the influence of compound 1, the number of entries into the dark arms was statistically significantly less than under the influence of diazepam in the studied dose (Table 8). Slow grooming was also more frequent with compound 1 (889%, P = 0.05).

Output. Compound 1 exhibited anxiolytic antiphobic activity in male rats in a cross-maze cross-labyrinth. At the same time, the antiphobic effect of compound 1 was more pronounced than that of the reference drug diazepam in the studied dose.

When studying the anti-anxiety activity of compound 1 according to the "open field" method [21], the following was recorded: 1) the latent period of exit from the center of the field; 2) horizontal motor activity - the number of crossed sectors; 3) vertical activity - the number of vertical posts; 4) research activity - the number of holes examined - "holes" (sniffing); 5) quick grooming - the number of acts of quick washing; 6) the number of boluses. The experiment took 2 minutes. On the basis of an increase in the latency period for leaving the center of the field, a decrease in vertical and horizontal activity, the “sedative” effect of the substance was judged, with an increase in exploratory activity, limitation of the frequency of fast grooming and the number of fecal boluses (vegetative representation of stress) - an “anti-anxiety” effect [22].

Evaluation of the effect of compound 1 on the behavior of male rats in the open field test revealed a statistically significant increase in exploratory activity and a decrease in the frequency of fast grooming, which indicated a decrease in the emotionality of the animals. Compared with the effect of diazepam against the background of the use of compound 1, the number of investigated "minks" was statistically significantly greater, the frequency of fast grooming significantly decreased (Table 9). Slow grooming also increased with the use of compound 1 (333%, P = 0.01).

Output. When tested in male rats in an open field, Compound 1 exhibited anti-anxiety effects. The anti-anxiety effect was more pronounced in comparison with the effect of diazepam in the studied doses.

Multivariate testing

In recent years, psychopharmacological methods for assessing the tranquilizing activity of psychotropic drugs have become widespread, based on the registration of the behavioral response of rodents exposed to stress. Along with the method of conflict situations, discriminatory tests are used. Each of the methods assesses the change in a certain behavioral response in animals, which is considered as a manifestation of anxiety, is reliably reproduced by stimuli that provoke anxiety in humans, and is suppressed by tranquilizers [11, 12].

For the purpose of a comprehensive study of the emotional behavioral status in animals, a multiparametric method for assessing anxiety-phobic states was used. The method takes into account the nature of the species-specific response of rodents to a series of ethologically adequate test stimuli that provoke manifestations of fear and anxiety, which do not cause the emergence of strong competitive motivations, easily reproducible with repeated repetition of experiments [23]. Multiparameter testing allows you to identify the main features of changes in behavioral activity and evaluate the anxiolytic effect when using methodological techniques, to get a systematic, holistic idea of the action of the test substance.

The tests of the multivariate method are based on situations that form responses to fear and anxiety in animals: 1) a collision with an unknown object ("inanimate") or a situation (1-4 tests) and 2) manipulations of the experimenter's hand (5-8 tests). The testing algorithm was the same: sequentially from 1 to 8 tests.

The first - the "step-down" test characterizes the instinctive-defensive behavior of the animal. Testing was carried out as follows: a 20 × 14 × 14 cm parallelepiped made of organic glass (box) was placed on a flat horizontal brightly lit surface. The rat was placed on the upper part of the box and the countdown of the time during which the animal descended to the floor with all four paws was started. In the case when the rat remained at the top of the box for 180 s, the action was considered not performed, the test was terminated, giving a score of 3 points.

In the second test, a two-chamber box with a size of 20 × 20 × 30 cm with a partition with a 7 × 10 cm hole in the center is used. paws.

The third testing was carried out using a "house", which is a transparent organic glass chamber with a size of 16 × 15 × 15 cm, one of the walls of which had an opening for the free exit of the animal from the enclosed space. The rat was placed in a "house", the hole was closed and placed in the center of a brightly lit area. After 15 seconds, the “house” was opened and the time of the animal's exit was recorded.

Research on tests 4-8 was carried out in an "open field".

In the fourth test, the occurrence of fear and anxiety in front of an open and unfamiliar space was assessed, associated with a decrease in motor activity. The animal was placed in the center of the field and the time during which the rat crossed the four central squares with all paws was recorded. If the animal did not leave the center of the field for 60 s, the action was considered not performed, the test was terminated, but observation was continued (5-8 tests).

The piging of the animals was assessed for the action of the experimenter's hand. Spontaneous boating and boating, caused by any external stimulus, is often observed in rats with a pathological feeling of fear and anxiety, which is a reliable indicator of the emotional state of the animal.

Tests 5 - 8 make it possible to assess the behavior of the animal on the action of the experimenter's hand: approaching and stroking. Testing was performed by slowly bringing the hand closer to the rat's face so that the animal could see the hand. Approximation and stroking were performed 2-3 times in a row. The intensity of the response was assessed in the range from 0 to 3 points (in the tests of the scale, discrete ranking was used with an interval of 1 point). The greater the severity of the response corresponded to the higher (in points) score, the absence of the response - the lower score, and, consequently, the lower anxiety-phobic level.

The answers were evaluated in points: according to tests 1-2: 0 - 0 <t <30 s; 1 - 30 <t <60 s; 2 - 60 <t <180 s; 3 - does not descend (does not pass) in 180 s. For test 3: 0 - 0 <t <15 s; 1 - 15 <t <30 s; 2 - 30 <t <180 s; 3 - does not go beyond 180 s. For test 4: 0 - 0 <t <15 s; 1 - 15 <t <30 s; 2 - 30 <t <60 s; 3 - does not leave for 60 s. In the case of fluctuations in the animal when performing tests 2 and 3 (incomplete passed the exit, peeking), the assessment was increased by 0.5 points. If the rat did not perform the actions according to tests 1-4, a score of 3 points was given. The intervals between tests 1-3 were 15-20 minutes.

According to tests 5-8, the absence of reactions of the animal in response to the "living object" was assessed at a minimum - 0 points. When a reaction to a contact stimulus (stroking) occurred, a smaller number of points was awarded - 1 than to a distant one (approaching a hand) - 2 points. The retention of these reactions (in case of occurrence) after the termination of the action of stimuli was assessed at the maximum - 3 points.

The total indicator for tests 1-4 (IDA - the index of physical activity) allows one to assess the "sedative" (increase in the indicator, decrease in physical activity) or activating (decrease in indicators, increase in physical activity) effect. The total score for tests 5-8 (index of emotional reactivity-IES) allows one to judge about the "anti-anxiety" (decrease in indicators, decrease in emotional reactivity) or "anxiogenic" (increase in indicators, increase in emotionality) effect of the test substance [22, 24].

Against the background of the use of compound 1 in male animals, an increase in the latent time of leaving the center of the field was noted, at the same time, the transitions through the opening of the box were accelerated. At the same time, the intensity of the reactions of stinging, hiding, vocalizing and pressing the ears significantly decreased, the index of emotional reactivity (IER) significantly decreased. In the group of animals treated with diazepam, the latent time of the tests was reduced in relation to the data of the control group, and the IDA was limited. Compared with the effect of diazepam against the background of the use of compound 1, the severity of the responses of animals when exposed to stimuli decreased, the indicator of emotional reactivity decreased (table 10).

Output. In multivariate testing of male rats, Compound 1 exhibited anti-anxiety effects upon single administration. The anti-anxiety effect was more pronounced in comparison with the effect of diazepam in the studied doses.

Psychostimulating activity

The psychostimulating effect of compound 1 was studied on sexually mature male white rats (Wistar lines, weighing 200–250 g, 7–10 in groups) in the lateral position test [11, 12]. The technique makes it possible to reveal the presence of a psychostimulating / hypnogenic effect of substances on the lateral position of animals against the background of high doses of hypnotic drugs. The technique is based on the ability of animals to remain in an “uncomfortable” position under the influence of high doses of hypnogenic substances. Animals of the control group, when turning over into an uncomfortable position for them on their backs, immediately reflexively return to their normal position. If the substances under study have a hypnotic effect, the animals remain in the "lateral position" - an uncomfortable position on the side or on the back. In the course of observation, researchers record the latency period of the lateral position and its duration.

1 , 0 g of active substance in a vial with a capacity of 20 ml; SINTEZ, JSC, Russia). Reference drugs were introduced similarly: caffeine sodium benzoate at a dose of 0.5 mg / kg (Coffeinum natrii benzoas; injection solution 100 mg / ml ampoule 1 ml; Novosibkhimpharm AO, Russia) and chlorpromazine at a dose of 0.7 mg / kg mg / kg (Chlorpromazinum; solution for injection 25 mg / ml, ampoules 1 ml; Novosibkhimpharm AO, Russia). Doses of the studied compounds and reference drugs were selected by the dose titration method and taking into account the literature data. Animals of the control group were injected with one thiopental sodium 30 mg / kg intraperitoneally.

When using a combination of compound 1 and thiopental sodium, the animals were not asleep (table 11). In addition, sleep duration was statistically significantly reduced. The effect was more pronounced than with the introduction of a comparison drug with the psychostimulating effect of caffeine sodium benzoate.

Conclusion: Compound 1 possesses psychostimulating activity exceeding the effect of sodium benzoate caffeine in the studied dose.

Acute toxicity

Determination of acute toxicity and calculation of LD ₅₀ in male and female mice were carried out according to the Behrens method and it was found that for compound (I) for female mice LD50 = 1526.85 mg / kg of animal body weight, for male mice LD ₅₀ = 750 mg / kg body weight of animals with intraperitoneal injection.

The LD _{50 of} metamizole sodium for mice is 250 mg / kg (intraperitoneally) [25]. LD _{50 of} lidocaine: for rats it was 317 mg / kg, for mice it was 220 mg / kg (orally) [26]. LD _{50 of} bromocriptine: for rats - 72 mg / kg (intravenously) [27]; bromocriptine mesylate: for rats - 10.5 mg / kg (intravenously), for mice - 189 mg / kg (intravenously) [28].

LD _{50 of} diazepam for mice is 700 mg / kg, for rats - 1200 mg / kg (orally) [29].

Thus, the test compound (1) is low-toxic, belongs to the IV class of toxicity according to K.K. Sidorov. (1973) [30].

Output. The research results show that the studied compound 1 has analgesic, anticataleptic, tranquilizing and psychostimulating activity.

List of sources

1. The Concise Guide to PHARMACOLOGY 2015/16: G Protein-Coupled Receptors. CGTP Collaborators [Text] / S. P. H. Alexander [et all] // Br. J. Pharmacol. - 2015. - Vol. 172. - P. 5744-5869. doi: 10.1111 / bph.13348.

2. Al-Hasani R. Molecular Mechanisms of Opioid Receptor-Dependent Signaling and Behavior / R. Al-Hasani, M. R. Bruchas // Anesthesiology. - 2011. - Vol. 115. - P. 1363-1381.

3. Simonson B. Pharmacology and anti-addiction effects of the novel κ opioid receptor agonist Mesyl Sal B, a potent and long-acting analogue of salvinorin / B. Simonson [et all] // Br. J. Pharmacol. - 2015. - Vol. 172. - P. 515-531. doi: 10.1111 / bph.12692.

4. Emelyanova T.G. Anti-inflammatory and analgesic activity of Tyr-Pro dipeptide and its analogues / TG Emelyanova [et al.] // Academic journal of Western Siberia. - 2014. - T. 10, No. 4 (53). - S. 88-89.

5. Guzman D.S. Evaluation of thermal antinociceptive effects and pharmacokinetics after intramuscular administration of butorphanol tartrate to American kestrels (Falco sparverius) / D. S. Guzman, T. L. Drazenovich, B. KuKanich, G. H. Olsen // American journal of veterinary research. - 2014. - Vol. 75, No. 1. - P. 11-18.

6. Codonidi I.P. Synthesis and biological activity of N-heterocyclic derivatives of 1,4-dihydro-4-oxopyrimidine / IP Codonidi [et al.] // Questions of biological, medical and pharmaceutical chemistry. - 2012. - No. 4. - S. 19-26.

7. Aminoquinazoline cannabinoid receptor modulators for treatment of disease and their preparation / Gahman, Timothy C .; Thomas, David J .; Lang, Hengyuan; Massari, Mark E. // PCT Int. Appl. (2008), WO 2008157500 A1 20081224.

8. Ovakimyan A.G., Bicherov A.A., Kodonidi I.P., Oganesyan E.T., Manvelyan E.A., Bicherov A.V., Zaichenko S.B., Tyurin R.V., Manvelyan M.M. Prediction, synthesis and study of dopaminergic activity of hydroxy- and methoxyphenyl derivatives of 4- (3) quinazolinone // Modern problems of science and education. - 2015. - No. 2-2. - S. 511.

9. Manvelyan E.A., Manvelyan M.M., Oganesyan E.T., Kodonidi I.P., Zaichenko S.B., Bicherov A.A., Bicherov A.V. 2-benzyl derivative of 4- (3 H) -quinazolinone, which has analgesic, antiparkinsonian, antihypoxant, tranquilizing effects. - Patent for invention No. 2715884. Date of issue: 24.01.2020.

10. Chervonskaya G.P., Pankratova G.P., Mironova L.L. Ethics of a biomedical experiment in preclinical research // Toxicological Bulletin. - 1998. - No. 3. - C. 2-8.

11. Guidelines for conducting preclinical studies of drugs. Part one / edited by A.N. Mironov. - M .: Grif i K, 2012 .-- 944 p.

12. Guidelines for experimental (preclinical) study of new pharmacological substances [Text] / ed. R. U. Khabrieva. - M., 2005 .-- 512 p.

13. Manvelyan E.A., Baturin V.A., Kolodiychuk E.V. Gender differences in the effectiveness of psychotropic and cardiotropic drugs: monograph. - Stavropol, 2016 .-- 130 p.

14. Glantz S. Biomedical statistics: [trans. from English]. - Moscow: Practice, 1999.

15. Rebrova O.Yu. Statistical analysis of medical data. Application of the STATISTICA application package. - Moscow: MediaSPHERE, 2006.

16. Grzhibovsky A.M., Ivanov S.V., Gorbatova M.A. Comparison of quantitative data of two independent samples using the STATISTICA and SPSS software: parametric and nonparametric criteria // Science and Health. - 2016. - No. 2. - S. 5-29.

17. Voronina T.A., Guzevatykh L.S. Methodological recommendations for the study of the analgesic activity of drugs // Guidelines for conducting preclinical studies of drugs. - M .: Grif and K, 2012 .-- S. 197-218.

18. Means with a local anesthetic effect Pat. 2313341 Rus. Federation: IPC A61K31 / 4188 / A61P23 / 02 / Galenko-Yaroshevsky A. P. [and others]; applicants and patent holders FGOU VPO "Southern Federal University", Non-state OU VPO "Kuban Medical Institute", FGU "Interdisciplinary Scientific and Technical Complex" Eye Microsurgery "named after acad. S.N. Fedorov "Fed. health agencies. and social development; No. 2006140600/15; Appl. 16. 11. 2006; publ. 12/27/2007. Bul. No. 38 - 8 p: ill.

19. Dubrovin A.N. [and etc.]. Synthesis and properties of 2-substituted amides of cinchoninic acid // Modern problems of science and education. - 2012. - No. 4. https://science-education.ru/ru/article/view?id=6630.

20. Rodgers R. J., Johnson N. J. Factor analysis of spatiotemporal and ethological measures in the murine elevated plus-maze test of anxiety // Pharmacol. Biochem. Behav. - 1995. - Vol. 52, N2. - P. 297-303.

21. Buresh J., Bureshova O., Houston J.P. - Techniques and basic experiments in the study of the brain and behavior. - Moscow: Higher school, 1991 .-- 398 p.

22. Manvelyan E.A., Anisimova N.A., Baturin V.Α. Sexual dissimilation of behavior under stress effects of varying intensity and changes in the function of the adrenal glands (monograph), Stavropol, 2013. - 107 p.

23. A multiparameter method for a comprehensive assessment of anxiety-phobic states in rats / V. I. Rodina [et al.] // Zhurn. higher. nerve. activities to them. I.P. Pavlova. - 1993. - No. 5. - S.1006-1017.

24. Oganesyan E.T., Kodonidi I.P., Sochnev V.S., Manvelyan E.A., Sysa V.Yu., Manvelyan M.M. A new N-arylsulfamide derivative of o-benzoylaminobenzoic acid with anxiolytic, actoprotective and antidepressant activity. - Patent for invention No. 2643356. Date of issue: 1.02.2018.

25. SAFETY DATA SHEET. Metamizole (sodium salt). Access mode: https://www.caymanchem.com/msdss/15771m.pdf (date of access 12/24/2018).

26. Toxicological Data on Ingredients: Lidocaine. Access mode: http: // www.sciencelab.com/msds.php?msdsId=9924492 (date of treatment 12/18/2018).

27. Material Safety Data Sheet. Bromocriptin. sc-337602. Access mode: http://datasheets.scbt.com/sc-337602.pdf (date of access 24.12.2018).

28. Material Safety Data Sheet. Bromocriptine (mesylate). sc-200395. Access mode: http://datasheets.scbt.com/sc-200395.pdf (date of access 24.12.2018).

29. Diazepam International Program on Chemical Safety Poisons Information. Access mode: http://www.inchem.org/documents/pims/pharm/pim181.htm (date of access 06.06.2018).

30. Guidelines for experimental (preclinical) study of new pharmacological substances / ed. VP Fisenko [and others] - M .: IAA Remedium, 2000. - 398 p.

The research was carried out with the support of the Fund for Assistance to Small Innovative Enterprises in Science and Technology (Fund for Assistance to Innovation; contract No. 12575GU / 2017 of 18.04.2018).

Table 1 - Influence of compound 1 on formalin test parameters in male rats (%)

Table 2 - The effect of compound 1 on the performance of the test "vinegar cramps" in male rats (%)

Table 3 - The effect of compound 1 on the performance of the test "vinegar cramps" in female rats

Substances M ± m / Δ% Control (distilled water), n = 12 28.17 ± 4.78 1, n = 10 9.0 ± 2.62 Δ% to control - 68.0 ** Δ% to lidocaine - 18.9 Lidocaine, n = 10 11.1 ± 3.89 Δ% to control - 60.6 *

Note - 100% - data from the control group of animals. Relative to the indicators of the control group of female rats and the comparison group: * - P <0.05, ** - P <0.01 (Student, Wilcoxon-Mann-Whitney, Kruskal-Wallis tests).

Table 4 - The effect of compound 1 on the formalin test parameters in female rats (at the stage of diestrus ½)

Substances Formalin test (M ± m / Δ%) All period Phase 1 (10 minutes) Phase 2 (50 minutes) Control 22.0 ± 14.79 12.0 ± 7.76 10.0 ± 7.05 1 3.0 ± 3.0 3.0 ± 3.0 0 ± 0 Δ% to control - 86.4 * - 75.0 - Δ% to metamizole - 81.2 (P = 0.057) - 58.3 - Δ% to lidocaine - 91.1 *** - 81.8 * - Metamizole sodium 16.0 ± 5.01 7.2 ± 2.22 8.8 ± 3.93 Δ% to control - 27.3 - 40.0 - 12.0 Lidocaine 33.75 ± 4.5 16.5 ± 3.18 17.25 ± 1.44 Δ% to control + 53.4 + 37.5 + 72.5

Notes - 100% - data of control groups of females in the stage of estrous cycle diestrus ½. Significantly relative to the group of female rats treated with the solvent, and the comparison groups in the phase of the estrous cycle diestrus ½: * - P <0.05; *** - P <0.001 (Student's, Wilcoxon-Mann-Whitney, Kruskal-Wallis tests).

Table 5 - The effect of compound 1 on the number of "vinegar writhing" in female rats in the stages of diestrus ½ and proestrus / estrus

Substances Stages of the estrous cycle The number of "vinegar writhing" (M ± m / Δ%) Control Diestrus ½ 28.29 ± 6.89 Proestrus / estrus 26.33 ± 6.08 1 Diestrus ½ 5.0 ± 3.52 Δ% to control - 82.3 * Δ% to lidocaine - 45.9 Proestrus / estrus 13.0 ± 3.26 Δ to control - 50.6 Δ% to lidocaine +5.4 Lidocaine Diestrus ½ 9.25 ± 6.5 Δ% to control - 67.3 * Proestrus / estrus 12.33 ± 5.28 Δ% to control - 53.2

Notes - 100% - data of control groups of females in the corresponding phases of the estrous cycle. Relative to the control group of female rats in the corresponding stages of the estrous cycle: * - P <0.05 (Student, Wilcoxon-Mann-Whitney, Kruskal-Wallis tests).

Table 6 - Effect of compound 1 on the cataleptogenic activity of haloperidol (c) in female rats

Table 7 - The effect of compound 1 on the cataleptogenic activity of haloperidol (c) in female rats at the stage of the estrus / estrus cycle

Table 8 - The effect of compound 1 and diazepam on the parameters of the behavior of rats in the elevated cruciform maze

Substances The number of entries into the illuminated arms Time in illuminated sleeves Number of visits to dark sleeves Time in dark sleeves Control group (K1) (solvent) n = 10 0.8 ± 0.42 10.3 ± 4.93 1.8 ± 0.29 289.7 ± 4.93 Compound 1; n = 9 0.56 ± 0.24 1.33 ± 0.88 1.22 ± 0.28 # 238.33 ± 35.24 Δ% to K 1 - 30.0 - 87.1 - 32.2 * - 17.7 ** Control group (saline) (K2) n = 7 0.86 ± 0.26 26.43 ± 8.57 1.29 ± 0.18 273.57 ± 8.57 Diazepam 0.1 mg / kg; n = 6 1.83 ± 0.17 66.67 ± 10.54 1.50 ± 0.22 233.33 ± 10.54 Δ% to K2 + 112.8 * + 152.3 * +16.3 - 14.3 *

Note - 100% - indicators of the behavior of male rats treated with a solvent (Tween-80 solution; K1 - a control group for animals receiving compound 1) and saline (K2 - a control group for rats receiving diazepam). * - the differences are statistically significant in relation to the data of the corresponding control group: * - at p <0.05; ** - at p <0.01; # - the differences are statistically significant in relation to the data of the group receiving diazepam (Wilcoxon-Mann-Whitney test).

Table 9 - Effect of compound 1 on the behavior of male rats in the "open field" with a single administration

Table 10 - The effect of compound 1 on the behavior indicators of male rats during multivariate testing

Table 11. Effect of Compound 1 on the behavior indicators of male rats in the "lateral position" test

Claims (3)

1. A method of obtaining 2- (3,4-dimethoxybenzyl-6-sulfo) quinazolin-4 ( 3H ) -one

corresponding to formula (1), in which Me is a methyl radical based on the reaction of 2- (3,4-dimethoxybenzyl) - ( 3H ) -quinazolin-4-one with concentrated sulfuric acid at room temperature, followed by the isolation of the reaction product upon dilution of the reaction mixture with water. 2. 2- (3,4-dimethoxybenzyl-6-sulfo) quinazolin-4 ( 3H ) -one corresponding to formula (1), having analgesic, antiparkinsonian, tranquilizing, psychostimulating activity.