RU2582966C2 - Means for reducing alcohol motivation in alcohol dependence - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular to pharmacology, and relates to a new use of a known substance hydrochloride of N-2-adamantyl-hexamethyleneimine (himantane) as a means for reducing alcohol motivation in alcohol dependence. It is shown that a single administration himantane rats were dose-dependently reduce the severity of alcohol-deprivation effect in chronic administration and reduces the average consumption of ethanol from the 3^rd day of the experiment. In addition, himantane hinders the realization of the acute effects of ethanol, blocking stimulation of motor activity and restoring incoordination with mild alcohol intoxication. None of the doses studied himantane not affect the duration of anesthesia in mice alcohol.

EFFECT: technical result consists in reducing alcohol motivation for alcohol dependence wherein himantane have no negative side effects such as the development of drug dependence and drug potentiation of ethanol effects.

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Description

The invention relates to medicine, in particular to pharmacology, and relates to a new use of the known substance of N-2-adamantyl-hexamethyleneimine hydrochloride (gimantan) as a means to reduce alcohol motivation in alcohol dependence.

Currently, strategies for the search, development and creation of new original drugs for the prevention and treatment of alcohol dependence continue to remain relevant for modern psychopharmacology and narcology, given the widespread and high socio-economic significance of this disease (Medical, social and economic consequences of drug addiction and alcoholism / E.A. Koshkina et al. - M .: Per SE, 2008 .-- 288 p.). The data on alcoholism published in the report of the Research Institute of Rosstat for 2011 show that in fact there are 12 million alcohol abusers in the country, while the arsenal of pharmacological agents for suppressing alcohol motivation and attraction is limited and is not very effective.

Excessive alcohol consumption over a long period of time can lead to psychological and mental disorders, as a result of a complex dynamic process that involves numerous adaptive changes affecting the neurobiological systems of “rewards” and responses to stress (Vengeliene V. et al., Br. J. Pharmacol. 2008; 154 (2): 299-315). Among the biological and environmental influences that form alcohol motivation, stress is considered as one of the most important factors determining alcohol consumption. Repeated repetition of “intoxication-withdrawal” cycles significantly changes the nature of the stress response, including dysregulation of the hypothalamic-pituitary-adrenal system (Heilig M, Koob GF. Trends Neurosci. 2007; 30 (8): 399-406). In recent years, much attention has been paid to the role of pathological dependence in terms of the formation of sustainable motivation and a return to alcohol consumption in the event of a breakdown in remission. This is due to the fact that the affective (psychological) components of the acute withdrawal syndrome and prolonged withdrawal symptoms seem to affect alcohol addiction to a greater extent than the physiological symptoms of alcohol deprivation (Heilig M. et al., Addict Biol. 2010; 15 (2): 169-84).

The disruption of the regulatory functions of the glutamatergic system is today considered as a key pathophysiological factor in the formation of alcoholism, as well as a mechanism that can be a target for pharmacotherapy of this disease (De Witte P. et al. Neuroscience and Biobehav. Rev. 2003; 27: 189-197; Spanagel R, Kiefer F. Trends in Pharmacol. Sci. 2008; 29: 109-115). Microdialysis has shown an increase in the extracellular glutamate content in modeling ethanol withdrawal syndrome in rats (Dahchour A, et al. Psychiatry Research. 1998; 82: 107-114), and this increase proportionally increased with subsequent “cancellation-withdrawal” cycles (Dahchour A, De Witte P. Alcohol Clin Exp Res. 2003; 27: 465-470). With the administration of a TBOA glutamate reuptake inhibitor, an increase in ethanol consumption is observed (Kapasova Z, Szumlinski KK Alcoholism: Clinical and Experimental Research. 2008; 32: 617-631), and vice versa, acamprosate, a functional antagonist of glutamate NMDA receptors, is currently approved a drug for treating alcoholism in the United States and several European countries (Spanagel R, Kiefer F. Trends in Pharmacol. Sci. 2008; 29: 109-115).

A decrease in neuronal activity, as well as a violation of the functional connection between the ventral striatum and the prefrontal cortex, is associated with an uncontrollable attraction to alcohol in patients with alcoholism in a state of withdrawal. In ethanol-preferred rats, against the background of alcohol deprivation, there is an increase in glutamatergic transmission in the dorsomedial striatum (DMS). In addition, exposure to ethanol and its abolition in ex vivo and in vivo experiments cause prolonged activation in the DMS of glutamate NMDA receptors containing NR2B subunits, contributing to an increase in the consumption of ethanol solution (Chen G. et al., Alcohol.Clin.Exp. Res. 2011; 35 (10): 1739-1748).

Table 1 presents experimental data on the action of two psychotropic drugs from the group of adamantane derivatives, the mechanism of action of which is associated with non-competitive blockade of NMDA receptors, in experimental models of alcoholism. Despite certain positive results achieved using memantine in animal experiments, the efficacy of memantine in alcohol-dependent patients has not been proven in pilot double-blind clinical trials (Evans SM et al .. Alcohol Clin Exp Res. 2007; 31 (5): 775-82). Further, using the method of nuclear magnetic resonance, it was shown that, compared with patients unburdened by the family history of alcoholism, people with a hereditary form of alcoholism are less sensitive to memantine, apparently due to the initially increased functional activity of NMDA receptors (Jamadar S. et al. Psychopharmacology (Beri). 2012 Jul; 222 (1): 129-40).

Using neuroimaging methods, neuroanatomical and neurochemical substrates of addictive behavior were identified. So, in the brain of a patient who is abusing alcohol, the amygdala is isolated, including the nucleus accumbens, the frontal orbit and the dorsal striatum, that is, the areas responsible for supporting effects, decision making and control of impulsive behavior. It was shown that a decrease in the functioning of the dopaminergic system correlates with a pathological attraction to alcohol and a breakdown in remission (Heinz A. et al., Am. J. Psychiatry. 2005; 162: 1515-15120).

           According to animal studies, with prolonged consumption of ethanol solutions, there is a decrease in dopamine synthesis and its release in the striatum against the background of alcohol withdrawal, which generally indicates a decrease in the functional activity of central dopaminergic neurons (Kaliszuk S. et al., Ann Univ Mariae Curie Sklodowska Med. 1990; 45: 163-7). Considering the fact that in vivo experiments, preference for ethanol and its excessive consumption are accompanied by dysfunction of the dopaminergic and serotonergic systems, evidence has now been obtained of the effectiveness of the simultaneous use of dopaminergic and serotonergic receptor agonists, in particular amphetamine and fenfluramine, to reduce the consumption of ethanol and prevent the occurrence of induced cancellation of ethanol seizures in laboratory animals (Mirovsky Y. et al .. Alcohol Clin Exp Res. 1995; 19 (1): 160-3; Yu YL et al., Alcohol. 1997; 14 (1): 45-8).

In the work of Cohen C. et al. (1999) irreversible MAO-B inhibitors pargilin and deprenyl weaken the reinforcing properties of ethanol and reduce the voluntary consumption of a 10% ethanol solution in rats, which also confirms the involvement of dopamine and serotonin in the control of alcohol behavior and suggests the use of MAO inhibitors that increase the content of these mediators due to the blockade of their metabolism during the pharmacotherapy of alcohol dependence (Cohen C. et al., Pharmacol Biochem Behav. 1999; 64 (3): 535-9).

Gimantan (N-2-adamantyl-hexamethyleneimine hydrochloride) is a new original drug created at the V.V. Research Institute of Pharmacology Zakusova ”RAMS based on the structure of adamantane, has a pronounced antiparkinsonian activity, proven in experimental models (Waldman EA and other Expert. And Klin. Farmakol. 1999; 4: 3-6; Kapitsa and other Psychopharmacol. And biol. narcology. 2005; 1: 822-827; Kapitsa I.G. et al. Exper. and Klin. Farmakol. 2011; 7: 9-12), and confirmed in the clinic in patients with initial stages of Parkinson's disease (Katunina E.V. . et al. Journal of Neurology and Psychiatry named after S.S. Korsakov. 2008; 108 (6): 24-27). In addition, in the model of the early (“domotor”) stage of Parkinson's disease, gimantan, like amantadine, prevents the development of cognitive impairment and depressive-like behavior in rats (Nepoklonov A.V. et al., Expert. And Klin. Farmakol. 2012; 11: 3-6). Table 2 presents experimental data on the complex mechanism of action of hematan on a number of neurotransmitter systems.

Thus, the presence in the spectrum of pharmacological activity of gimantan of antidepressant and anti-anxiety actions, a complex mechanism of action, including a dopaminergic component, blockade of the ion channels of the glutamate receptors of the NMDA subtype and inhibition of MAO-B, as well as the absence of negative side effects, such as the development of drug dependence and drug drug potentiation the effect of ethanol, allow it to be used as a means to reduce alcohol motivation in alcohol dependence. In the available literature to date, there are no indications of the use of gimantan for the proposed purpose.

This study was performed on male mice of the CD-1 line with a body weight of 20-22 g. (Nursery RAMS "Pillar", n = 109), inbred male mice of the DBA / 2 line with a body weight of 20-22 g. (Nursery RAMS “Pillar”, n = 41), outbred male rats (cattery RAMS “Pillar”, n = 55) at the age of 8 months with an average weight of 470-490 g in the active phase of the experiment.

Gimantan (N-2-adamantyl-hexamethyleneimine hydrochloride, a substance synthesized at the VV Zakusov Research Institute of Pharmacology RAMS RAMS) in effective doses of 5.0, 10.0 and 20.0 mg / kg was dissolved in water for injection and administered intraperitoneally (in / b) at the rate of 0.1 ml / 10 g (mouse) or 0.1 ml / 100 g (rat) of the mass of the animal.

Methodology for the formation of alcohol dependence in outbred rats.

With the exception of a few selective inbred lines, rodents have an inborn aversion to the taste of alcohol and usually avoid consuming it. However, rats and mice can be “trained” to drink relatively large volumes of alcohol using an ethanol solution with the addition of sucrose, which corresponds to a typical pattern of human alcohol behavior, when addiction to alcohol begins with the consumption of sweet alcohol-containing cocktails with a gradual transition to unsweetened drinks. Another way of forced alcoholization of animals is to provide an ethanol solution as the sole source of liquid, followed by an assessment of the level of alcohol motivation in the conditions of free choice between ethanol and water (Anokhina I.P. and Kolik L.G. Guidelines for preclinical studies of drugs. Part one . - M.: Grif and K, 2012 .-- S. 310-333).

To model the pathological attraction to ethanol, we used the method of forced alcoholization (stage 1) of animals with the provision of a 10% solution of ethanol as the only source of fluid for 20 weeks. The animals were kept in individual cages under standard vivarium conditions of the FSBI “V.V. Zakusova »Frames with the provision of briquetted ad libitum feed with an adjustable 12/12 (light / dark) light mode. Once a week, changes in body weight of animals and the amount of ethanol consumed (in g / kg) were recorded. In the process of forced alcoholization, rats consumed physiologically significant amounts of alcohol (4.93 ± 0.09 g / kg in terms of ethanol). Further, in order to simulate the “intoxication - withdrawal” cycles observed in clinical practice, the method of voluntary alcoholization (stage 2) was used, giving the animals a choice between a vessel with 10% ethanol and a vessel with water for 24 hours per day with repeated periods of 48-hour alcohol deprivation (21st, 23rd and 25th weeks from the moment of first contact with ethanol). As a result of multiple "withdrawal syndromes", the alcoholic behavior of animals, previously considered as "controlled", is gradually transformed into "uncontrolled", which allows transmitting the obtained data to the human population, in which compulsiveness is a distinctive feature of alcoholism, that is, a strong addiction and almost painful craving for alcohol (Spanagel R. Physiol. Rev. 2009; 89: 649-705).

The animals were divided into experimental and control groups, taking into account body weight, the amount of alcohol consumed, the severity of the ethanol withdrawal syndrome, and were kept under standard vivarium conditions in individual cells until the start of the experiment.

Estimated average daily ethanol solution intake.

The most common method for studying the reinforcing properties of ethanol in laboratory animals when modeling alcohol dependence is the measurement of the amount of liquids consumed while providing unlimited access to a vessel with an ethanol solution of a selected concentration and a vessel with water for 24 hours a day.

Assessment of the level of alcohol motivation is carried out according to the average daily ethanol consumption (based on 1 kg of animal weight per day). Since the effort required to obtain ethanol is minimal, this technique allows you to evaluate only the motivational component of alcohol behavior (Tabakoff B. and Hoffman P. L. Alcohol Res Health. 2000; 24 (2): 77-84).

The experiments were performed on rats consuming relatively large amounts of 10% ethanol (4.00 ± 0.29 g / kg per day). The effect of gimantan in an effective dose of 20.0 mg / kg / day on voluntary alcohol consumption by animals with stable alcohol motivation, formed during 20 weeks of forced alcoholization, was evaluated. The daily alcohol consumption of individual animals was measured, then the average daily alcohol consumption (in g / kg) in the corresponding group of animals was calculated and the background consumption data (before injection) was compared with the data on the consumption of the ethanol solution by the animals while using the study drug.

Estimation of alcohol consumption in ADE conditions.

Alcohol-deprivation effect (ADE) - the reaction of animals to the repeated provision of free access to alcohol during the withdrawal period in the form of a temporary increase in its consumption compared to the previous period of alcohol deprivation.

The model allows to quantify the level of voluntary alcohol consumption after different periods of deprivation of animals access to ethanol: at the stage of formation of alcohol motivation or at the stage of development of physical dependence on ethanol in the conditions of free choice between ethanol and water.

During the deprivation period, animals in individual cages equipped with drinking bowls with an ethanol solution and water were deprived of access to the ethanol solution. 48 hours after ethanol withdrawal, the animals were again provided with an ethanol solution, after which the assessment of individual fluid intake was carried out under the same conditions as the assessment of voluntary alcohol consumption in the conditions of free choice between ethanol solution and water. The amount of liquids in the drinkers after providing ethanol is measured: 90 minutes after the start of the individual test; 24 hours after the start of an individual test; with a test lasting more than a day - every subsequent 24 hours. The amplitude of ADE was estimated as the difference between the rate of alcohol consumption in the first 90 minutes after deprivation and those before deprivation, as well as the rate of recovery of alcohol consumption to the level before deprivation of alcohol. The alcohol consumption was determined and expressed in g / kg.

Assessment of the effect of the studied substance on the ethanol-induced hyperlocomotor reaction (test "ethanol-induced stimulation of motor activity").

This test is used to identify the pathophysiological mechanisms underlying the formation of the addictive state. In mice, ethanol in low and moderate doses stimulates motor activity, which, according to some authors, mimics alcohol-induced euphoria in humans (Wise RA and Bozarth MA Psychol. Rev. 1987; 94: 469-492; Koob GF TIPS, 1992 ; 13: 177-184).

The registration of motor activity was carried out in an ORTO-VARIMEX 4 actometer (Columbus Instruments, USA) equipped with 8 pairs of IR sensors at a height of 1.27 cm from the floor, located along the perimeter of 40 × 40 cm of the experimental setup, under natural light for 10 minutes. Immediately after the introduction of ethanol (20% solution at a dose of 2.0 g / kg, volume 20 ml / kg body weight, ip), the mice were placed in OPTO-VARIMEX 4 and the total distance (in cm) covered by the animals was automatically recorded. When assessing the effect of gimantan at a dose of 20.0 mg / kg on the acute stimulating effect of ethanol, the drug was administered 30 minutes before ethanol injection.

Evaluation of the influence of the studied substance on muscle relaxation induced by ethanol (test "rotating rod").

The experiments were performed on outbred male mice. The installation is a rod 6 cm in diameter, raised to a height of 60 cm, with a fixed rotation speed (6.5 rpm). When assessing the effect of pharmacological substances on ethanol-induced ataxia, mice were pre-tested at least three times (each exposure time is 3 minutes) to achieve stable performance. Ethanol was administered (20% solution at a dose of 2.0 g / kg, volume 20 ml / kg body weight, ip) 45 minutes before testing. The main recorded indicator is the retention time on the rod (Rustay N.R. et al. Proc Nati Acad Sci USA. 2003; 100 (5): 2917-22.4). The ability of animals to maintain equilibrium and increase the residence time on a rotating rod under the action of the studied substance, introduced against the background of ethanol, is regarded as the ability to reduce the toxic effect of ethanol and exert a sobering effect. Conversely, a decrease in the retention time on a rotating rod under the action of the studied substances indicates an increase in muscle relaxation induced by ethanol. Gimantan in the range of effective doses of 5.0, 10.0 and 20.0 mg / kg was administered 30 minutes before testing (against the background of the action of a test dose of ethanol).

Assessment of the influence of the studied substance on the duration of sleep caused by ethanol (test "ethanol anesthesia")

The experiments were performed on outbred male mice. The animals were injected with a 25% solution of ethanol in a test dose (5.5 g / kg, ip), which caused the development of alcohol anesthesia, which lasts for 1-2 hours, which allows one to detect both a weakening and an increase in the action of ethanol. Animals were placed on a flat horizontal surface and the time of onset and end of alcoholic anesthesia was recorded by adopting a “lateral position” and, accordingly, an independent sustainable exit from it. The studied substance was administered against the background of ethanol anesthesia. The control group of animals was injected with a solution of ethanol in a / b test dose and an isomeric amount of the solvent used for the test substance (water for injection). The volume of injected 25% solution of ethanol (V) in accordance with its required dose (D) taking into account the weight of the animal (M) was calculated by the formula:

V (ml) = (D (g / kg) × M (g)) / 250.

The test "ethanol anesthesia" is used for a total assessment of the activity of ethanol-metabolizing systems and the sensitivity of the central nervous system to ethanol.

Statistical processing of the results was carried out using a one-way ANOVA analysis of variance followed by the Duncan test and Student t-test. The graph data are presented as mean values and standard error of mean values ± SEM.

The invention is illustrated by the following examples.

Example 1. The effect of gimantan on the alcohol deprivation effect (ADE) in rats with sustained alcohol motivation.

The experiments were performed on outbred male rats (n = 37) with an alcohol dependence formed over a period of 6 months. When modeling the state of acute alcohol withdrawal, it was found that gimantan in the dose range from 5.0 to 20.0 mg / kg with a single systemic administration reduces the severity of the alcohol deprivation effect in a dose-dependent manner. The maximum activity of the drug compared with the control group (p <0.001) appears at a dose of 20.0 mg / kg (Fig. 1). A similar action has previously been described for non-competitive NMDA receptor antagonists such as acamprosate, memantine, and neramexane (Spanagel R., Physiol. Rev. 2009; 89: 649-705).

A subsequent evaluation of the effect of hematan on the restoration of ethanol consumption in the first 24 hours after the alcohol deprivation effect in rats with stable alcohol motivation revealed a tendency to a decrease in ethanol consumption, especially at doses of 10.0 and 20.0 mg / kg (Fig. 2), which allowed the use of the drug in further studies with a chronic route of administration, it is at a maximum dose of 20.0 mg / kg.

Example 2. The effect of gimantan on the average daily consumption of ethanol.

The experiments were performed on outbred male rats (n = 18) with an alcohol dependence formed during 6 months (consumption 4.00 ± 0.29 g / kg / day in terms of pure ethanol). The effect of gimantan at the most effective dose of 20.0 mg / kg in chronic systemic administration on the average daily intake of a 10% ethanol solution was studied under the conditions of free choice between ethanol and water for 24 hours a day. It was shown that gimantan reduces the amount of ethanol consumed by animals, compared with the control group during the entire period of administration of the drug. A statistically significant effect develops from the 3rd day of the experiment (Fig. 3).

Example 3. The effect of hematan on the stimulation of motor activity caused by ethanol.

The experiments were performed on intact inbred male mice of the DBA / 2 line (n = 41), characterized by increased sensitivity to the stimulating effect of ethanol. During preliminary testing of mice in an actometer, it was shown that the basic level of motor activity of animals from the control and experimental groups does not differ. With acute administration of ethanol at a dose of 2.0 g / kg, ip, caused a significant increase in motor activity in animals from the control group compared with the background value (p <0.001). Gimantan at a dose of 20.0 mg / kg, without affecting the level of motor activity per se, statistically significantly (p <0.01) prevented the formation of an ethanol-induced hyperlocomotor reaction (Fig. 4).

The results are consistent with data on the ability of antagonists of NMDA receptors MK-801 and ifenprodil to block the stimulatory effect of ethanol in DBA / 2J mice (Broadbent J. et al., Psychopharmacology (Berl). 2003; 167 (3): 225-34).

Example 4. The effect of hematan on ataxia caused by ethanol.

The experiments were performed on outbred male mice (n = 68). In contrast to diazepam, a tranquilizer traditionally used to correct disorders related to alcohol dependence and causing increased muscle relaxation induced by ethanol in the “rotating rod” test, gimantan in doses of 10.0 and 20.0 mg / kg in a dose-dependent manner restored impaired coordination of movement and increased the retention time on the “rotating rod” on the background of the action of ethanol compared with the control group (p <0.05, p <001), which indicates the effectiveness of gimantan in light alcohol intoxics nations (Fig. 5).

Example 5. Evaluation of the effect of hematan on the duration of ethanol anesthesia.

The experiments were performed on outbred male mice (n = 41). When injecting a narcotic dose of ethanol 5.5 g / kg, the hypnosedative effect of ethanol is observed, which manifests itself in an increase in the time spent in the lateral position compared to background values (p <0.05). Gimantan in none of the studied doses of 10.0 and 20.0 mg / kg had a statistically significant effect on the duration of ethanol anesthesia, which allows us to conclude that there is no effect of gimantan on the system of ethanol-metabolizing enzymes (Fig. 6), which distinguishes the drug from another non-competitive memantine NMDA receptor antagonist, which, like benzodiazepine tranquilizers, potentiates the duration of alcohol sleep (Beleslin DB et al .. Alcohol. 1997 Mar-Apr; 14 (2): 167-73).

Based on the data obtained, it can be concluded that gimantan is an effective tool to reduce the craving for alcohol, since the drug: 1) with a single administration weakens the craving for alcohol in terms of alcohol deprivation effect; 2) with chronic administration for 2 weeks, it reduces alcohol motivation, reducing the average daily ethanol consumption in animals with a developed alcohol dependence. In addition, gimantan prevents the realization of the acute effects of ethanol, blocking the stimulation of motor activity and restoring impaired coordination of movements with mild alcohol intoxication.

Description of the drawings.

FIG. 1. The effect of gimantan on the alcohol deprivation (ADE) effect in rats with sustained alcohol motivation.

The abscissa axis shows the background values of ADE before the introduction of hematan (1) and experimental (2) data, the ordinate axis shows the amount of ethanol consumed (in g / kg) for 90 minutes after 48-hour alcohol deprivation. Experimental groups: 1st column — control group (water for injection), 2nd column — hemantan at a dose of 5.0 mg / kg, 3rd column — hemantan at a dose of 10.0 mg / kg, 4th column — hemantan at a dose 20.0 mg / kg.

The ordinate indicates the amount of alcohol consumed in 90 minutes (in g / kg). ** - p <0.01, ** - p <0.001 statistically significant differences compared with the control group, ANOVA, Duncan test; the number of animals in the group is 9-10; mean ± SEM.

FIG. 2. The effect of gimantan upon a single administration on the restoration of ethanol consumption in the first 24 hours after the alcohol deprivation effect in rats with stable alcoholic motivation.

The abscissa shows experimental groups: 1st column — control group (water for injection), 2nd column — hemantan at a dose of 5.0 mg / kg, 3rd column — hemantan at a dose of 10.0 mg / kg, 4th column - gimantan in a dose of 20.0 mg / kg.

The ordinate indicates the amount of alcohol drunk 24 hours after 48-hour alcohol deprivation (in g / kg).

There are no statistically significant differences compared with the control group; ANOVA; the number of animals in the group is 9-10; mean ± SEM.

FIG. 3. The effect of gimantan at a dose of 20.0 mg / kg in chronic systemic administration on the average daily intake of ethanol.

The abscissa shows the days of the experiment from 0 (background values before injection) to 14, when the amount of consumed fluids was measured. Experimental groups of animals: solid line with a square marker - water for injection (control); dashed line with a round marker - gimantan in a dose of 20.0 mg / kg (experiment).

The ordinate indicates the amount of ethanol consumed per day (in g / kg).

*** -p <0.001, ** -p <0.01 statistically significant differences compared with the control group according to the Student t-test, the number of animals in each group is 9, mean ± SEM.

Figure 4. Effect of hematan on ethanol-induced hyperlocomotor response in DBA / 2 mice.

The following groups of animals are represented along the abscissa: 1st column — water for injection (background); 2nd column - ethanol at a dose of 2.0 g / kg (control); 3rd column - gimantan in a dose of 20.0 mg / kg; The 4th column is ethanol at a dose of 2.0 g / kg + gimantan at a dose of 20.0 mg / kg.

The y-axis is the distance traveled in cm.

*** - p <0.001, ** - p <0.01, statistically significant differences compared with the control group according to the ANOVA univariate analysis of variance, Duncan test, the number of animals in each group is 9-12, mean ± SEM.

Figure 5. The effect of hematan on muscle relaxation induced by ethanol in the test "rotating rod" in mice of the line CD-1.

On the abscissa axis of groups of animals that received: a white column - water for injection (background); column with vertical hatching - ethanol in a dose of 2.0 g / kg (control); a column with a horizontal hatching - ethanol + gimantan in a dose of 5.0 mg / kg; column with a diagonal hatching - ethanol + gimantan in a dose of 10.0 mg / kg; the black column is ethanol + gimantan in a dose of 20.0 mg / kg.

On the y-axis - the equilibrium retention time on a rotating rod in seconds (sec).

* -p <0.05, ** - p <0.01 statistically significant differences compared with the control group according to the univariate analysis of variance ANOVA, Duncan test; the number of animals in each group is 12-18; mean ± SEM.

6. Evaluation of the effect of hematan on the duration of ethanol anesthesia in CD-1 mice.

The abscissa represents the groups of animals that received: 1st column - water for injection (background); 2nd column — ethanol at a dose of 5.5 g / kg (control); 3rd column - ethanol at a dose of 5.5 g / kg + gimantan at a dose of 10.0 mg / kg; The 4th column is ethanol at a dose of 5.5 g / kg + gimantan at a dose of 20.0 mg / kg.

On the y-axis - the time spent in a lateral position in minutes (min).

* -p <0.05 statistically significant differences compared with background values, according to ANOVA, Duncan test; the number of animals in each group is 10-11; mean ± SEM.

Claims (1)

The use of N-2-adamantyl-hexamethyleneimine hydrochloride (gimantan) as a means to reduce alcohol motivation in alcohol dependence.

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