RU2508098C1 - Anticonvulsant and method for preparing it - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, particularly to pharmacology and pharmaceutics, and concerns an anticonvulsant representing the amino acid glycine immobilised on detonation nanodiamond particles 2-10 nm in size, and to a method for preparing it.

EFFECT: improved efficacy of the drug preparation.

4 cl, 5 dwg, 7 tbl, 3 ex

Description

The invention relates to medicine, in particular to pharmacology, and relates to an anticonvulsant, which is glycine immobilized on particles of detonation nanodiamonds.

Epilepsy is the second most common neurological disease after a stroke [1]. It represents a heterogeneous symptom complex, a chronic disease characterized by the periodic occurrence of seizures. Their causes may be a large number of diseases with neurological manifestations [1]. About 50 million people (up to 1% of the world's population) suffer from epilepsy. Her drug therapy should provide effective control of seizures, but also maintaining a good quality of life, which is largely determined not only by the effectiveness, but also by the safety of treatment. It is known that in about 25% of patients, convulsions are not controlled by the main anticonvulsants. Moreover, monotherapy of epilepsy and other paroxysmal conditions is ineffective in 20-30% of cases. As a rule, combinations of anticonvulsants are used [1-3]. This, in turn, increases the risk of various, including serious side effects, which reduces the quality of life of patients. Therefore, the search and development of new low-toxic agents for the prevention and treatment of epilepsy and other paroxysmal conditions is an extremely urgent task.

It is known that the replaceable amino acid glycine (NH ₂ CH ₂ COOH), being the central neurotransmitter of the inhibitory type of action, exhibits a sedative effect and improves metabolic processes in brain tissues [2]. In modern therapeutic practice, glycine is used as a means of reducing the craving for alcohol, reducing withdrawal symptoms, depressive disorders, increased irritability, and normalizes sleep, as well as in the complex treatment of cerebrovascular disorders [2]. The pharmacological action of glycine is based on the effect of amplification of metabolic and neurotransmitter processes that occur due to an increase in its endogenous synthesis. It is possible to increase the intracellular synthesis of glycine only using signal transmission pathways due to interaction with receptor systems. Its interaction with glycine receptors leads to the opening of chlorine channels, hyperpolarization of the membrane and the spread of inhibition. Along with this, glycine is able to act as an allosteric coagonist of glutamate receptors. By binding at a specific site, it enhances the ability of glutamate and N-methyl-D-asparagus (NMDA) to open the cation channel [4, 5].

The pharmacopoeial glycine used is prescribed in the form of tablets (0.1 g each) under the tongue 3-4 times a day.

Known glycine immobilized on particles of detonation nanodiamonds with a size of 2-10 nm, used as a binder component in polymer composites [6, 7]. The method of obtaining it is as follows [7]. A portion of the nanodiamond is placed in a constant helium current reactor and annealed at a temperature of 150-470 ° C for 3-4 hours. Then, the nanodiamond samples are fluorinated at a temperature of 50-500 ° C for 1-24 hours by contacting with a mixture of gaseous fluorine and hydrogen . To obtain glycine immobilized on nanodiamond particles, fluorinated nanodiamonds are sonicated in o-dichlorobenzene for 20-30 minutes, glycine ethyl ester hydrochloride (NH ₂ CH ₂ COOH ₂ CH ₃ · HCl) and a few drops of pyridine are added. The resulting mixture was stirred at a temperature of 130-140 ° C for 8-12 hours. The resulting product was filtered, washed with ethanol and dried under vacuum at 70 ° C.

An additional characteristic of this substance is the size of its particles in suspension, equal, according to the data of dynamic light scattering (DLS), 310 nm [7].

A feature of this substance is the presence on the surface of nanodiamond particles, in addition to glycine molecules, also fluorine atoms. Although the authors declare their number to be less than 1% at., In reality it has been experimentally established that the fluorine concentration on the surface of a nanodiamond can reach 14% at. and more. This is due to the fact that the CF bond (E _st = 115 kcal / g-atom) is strong and the fluorine derivatives of carbon are inert with respect to many substances. Therefore, during the chemical immobilization of glycine on the surface of a nanodiamond containing fluorine atoms, glycine molecules are replaced by fluorine atoms only partially. Moreover, it is known that the presence of fluorine and its derivatives in the organic matter increases its toxicity and can change the indices of the microsomal xenobiotics biotransformation system in the liver [8]. Thus, the presence of fluorine atoms in the nearest nanostructured analog of nanodiamond, fullerene (C ₆₀ ), increases its general toxicity by 2.4–5 times [9]. In addition, fluorine and its compounds are able to accumulate in various environmental objects and be present in them in various quantities [10]. Therefore, glycine immobilized on nanodiamond particles that contain fluorine atoms is undesirable in medical practice as a medicine.

Obtaining glycine immobilized on particles of detonation nanodiamonds that do not contain fluorine atoms, with an increased dispersion in a suspension used as a medicine, as well as reducing the level of environmental and endoecological danger, simplifying and cheapening the method for its preparation, is an urgent and practically significant task.

The aim of the invention is to increase the effectiveness of the anticonvulsant effect of glycine without manifesting any side and toxic effects and expanding the range of anticonvulsant drugs.

The use of glycine immobilized on detonation nanodiamond particles as an anticonvulsant is not described in the scientific and patent literature.

The goal is achieved using the anticonvulsant described in accordance with the invention, which is glycine immobilized on particles of detonation nanodiamonds with a size of 2-10 nm, not containing fluorine atoms and having a shell thickness of up to 1 nm, with a glycine content of up to 21 ± 3 wt%. , and how to obtain it.

The described anticonvulsant in the form of glycine immobilized on particles of detonation nanodiamonds that do not contain fluorine atoms on its surface is an ultrafine powder (Figure 1) of a dark gray or dark gray color with a greenish or dark blue shade with a particle size of 2 to 10 nm, having a shell with a thickness of up to 1 nm (Figure 2), the size of the aggregates in an aqueous suspension of up to 100 nm (Figure 3) and a glycine content of up to 21 ± 3% by weight, which is part of the surface shell.

Figure 1 clearly shows the presence of the described anticonvulsant agent of an ultrafine structure of particles with a size smaller than the resolution of the used device (from 20 nm).

A micrograph of the particles of the described anticonvulsant was obtained on a Zeiss Ultra Plus field emission scanning electron microscope (Carl Zeiss, Germany).

Figure 2 shows that the particle size of the described anticonvulsant, coated with a thickness of up to 1 nm, is 2-10 nm.

A micrograph of the particles of the described anticonvulsant was obtained with a Jeol 1011 transmission electron microscope (JEOL, Japan).

In figure 3. the particle size distribution curve in the suspension of the described anticonvulsant is shown, from which it follows that the particle sizes in the suspension do not exceed 100 nm.

The particle size distribution of the described anticonvulsant in suspension was measured by DLS on a ZetaSizer instrument (Malvem Instruments, USA). The abscissa represents the logarithmic scale of particle size in nm. The ordinate axis is the percentage of particles with specific sizes.

The elemental composition of the particle surface of the described anticonvulsant according to X-ray photoelectron spectroscopy (XPS) is given in Table 1.

Table 1 Elemental composition and binding energies of surface atoms of the described anticonvulsant Name of characteristic Chemical elements C O N Atomic% 77.5-94.5 4-14 1,5-8,5 Communication energies, eV 285.2 ± 0.5 530.7 ± 0.5 399.8 ± 0.5

The surface of the described anticonvulsant was studied on a LAS-3000 instrument (Riber, France) equipped with an OPX-150 hemispherical analyzer. To excite photoelectrons, nonmonochromatized x-ray radiation of an aluminum anode (AlK _α = 1486.6 eV) was used at a tube voltage of 12 kV and an emission current of 20 mA. Calibration of the photoelectron peaks was carried out along the C 1s carbon line with a binding energy of 285 eV. The vacuum in the working chamber was 6.7 · 10 ^-8 Pa. To obtain a high vacuum, an ion pump was used.

The amount of glycine in the described anticonvulsant is determined as follows. Mixtures of nanodiamonds with different glycine contents are prepared. Take samples of each mixture of equal weight. Their PC spectra are recorded, the most intense characteristic signals are selected on them, which are correlated with the IR spectrum bands of the initial glycine. Then build calibration curves of the dependence of the signal intensity in the IR spectrum on the glycine content in the sample. Further, according to the intensity of the selected characteristic bands of the investigated anticonvulsant, the quantitative glycine content in it is determined from the calibration curves. According to the data obtained, the average value of the glycine content in the described anticonvulsant is determined.

The described method of obtaining an anticonvulsant is as follows. Detonation nanodiamonds are annealed in a stream of hydrogen gas at a temperature of 500-1200 ° C for 1-8 hours, then subjected to liquid-phase chlorination with molecular chlorine by photochemical treatment with visible light at a temperature of 50-70 ° C for 36-60 hours, followed by washing with carbon tetrachloride by centrifugation and drying under vacuum. Chlorine-modified nanodiamonds are dissolved in a polar solvent to form a suspension. Tertiary amine and glycine are added and the resulting mixture is sonicated for 5-60 minutes, followed by exposure at 50-80 ° C for 12-48 hours, centrifugation, washing with solvent and drying. Ultrasound treatment is carried out for 5-60 minutes, triethylamine is used as a tertiary amine and pyridine, a lower aliphatic alcohol, a water-alcohol mixture or water are used as a polar solvent.

The described anticonvulsant does not contain fluorine atoms (Table 1), as well as atoms of other halogens, in an amount exceeding the error of the device (0.1% at.), Since in the process of obtaining an anticonvulsant, all chlorine atoms are replaced by glycine molecules and leave from the surface of a nanodiamond in the form of HCl molecules.

In the process of pharmacological studies, work was carried out to study the specific anticonvulsant effect of the described drug and its harmlessness in comparison with pharmacopoeial glycine.

Evaluation of the anticonvulsant effect was carried out according to the Guidelines for the study of the pharmacological activity of substances described in [11]. For this, two methods were used with a different mechanism for the occurrence of seizures: electroshock and chemical exposure. The technique of antagonism with seizures caused by the maximum electric shock (exposure to electric shock) simulates seizures of the Grand Mal type. The antagonism technique with corazole (chemical exposure) simulates seizures of the Petit Mal type. These methods are basic models in the search and study of substances with anticonvulsant activity.

The study of the anticonvulsant activity of the described drug was carried out on outbred white mice males weighing 24-28 g.

To obtain seizures, we used a certified device - Rodent Shocker RS, type221 (Harvard Apparatus, GmbH, Germany). After the maximum electroshock, the animal developed tonic extension of the fore and hind limbs, ending in death. The ability to prevent the development of tonic extension and reduce the death of animals was evaluated.

It was found that conducting maximal electroshock in control animals caused tonic extensia, ending in the death of 80% of mice, and only 20% of the animals survived after electroshock (Table 3). The described anticonvulsant at a dose of 10 mg / kg reliably protected 60% of mice from death, and at a dose of 15 mg / kg - 70% of mice after maximal electroshock (Table 3). Pharmacopoeial glycine in doses of 10 and 15 mg / kg did not affect seizures and did not eliminate the death of mice with maximum electroshock - all animals showed tonic extension and subsequent death (Table 3).

Thus, the described anticonvulsant at a dose of 10-15 mg / kg has a distinct anticonvulsant effect in the act electric shock test, statistically reliably preventing the development of tonic extension and death of animals.

To study the anticonvulsant activity of the described agent in the antagonism test, with chemical exposure, corazole was used, which was injected subcutaneously into the cervical region of the back. 40 minutes before the administration of corazole, the described agent was introduced and distilled water as a control.

It was found that the described agent at a dose of 15 mg / kg caused a statistically significant increase (1.54 times) in the latent period of the appearance of the first generalized clonic seizures and an increase (2.64 times) in the life expectancy of animals compared to the control (Table 4) . Along with this, the described tool significantly reduced by 2 times the number of dead animals (table 4). Pharmacopoeial glycine at a dose of 15 mg / kg did not exert anticonvulsant effect in the antagonism test with corazole (Table 4).

The data obtained indicate the presence of the described anticonvulsant at a dose of 15 mg / kg of pronounced anticonvulsant activity in the antagonism test with corazole and the absence of an anticonvulsant effect in pharmacopeia glycine at a dose of 15 mg / kg.

A study of the acute toxicity of the described anticonvulsant was conducted according to the Guidelines for the study of acute toxicity described in [12].

During the experiment, the following indicators were recorded: the nature of the coat, changes in the condition of the mucous membranes, ptosis of the upper eyelid, increased urination, increased bowel movements, increased salivation, piloeraction, vocalization, lateral position, rhythm and depth of respiratory movements, aggressiveness, timidity, tremor, convulsions, change in thresholds of pain reaction, change in posture, catalepsy, impaired coordination of movements in the test of a rotating rod, holding for 5 seconds on an inverted mesh platform, erelezanie with inverted mesh platform up, the presence of pineal, corneal reflex, sedation, stereotypes and grooming, the death of the animal.

Statistical processing of the results was carried out using statistical packages “BioStat” for Windows. Group averages and standard error indicators were calculated.

The obtained results convincingly show that the described anticonvulsant when administered intraperitoneally to mice at doses of 75, 150 and 225 mg / kg, as well as pharmacopoeial glycine did not cause signs of intoxication and death of animals over 14 days. observations.

As a result of the pharmacological study, it was found that the described anticonvulsant has a pronounced specific anticonvulsant effect superior to the anticonvulsant effect of pharmacopeia glycine, and the absence of side effects and toxic effects in a dose exceeding the therapeutic by more than 20 times.

The described anticonvulsant not only improves the therapeutic efficacy of the pharmacopeia drug glycine, but also expands the range of effective and safe anticonvulsants.

A brief description of the graphic materials.

figure 1. An electron micrograph of the described anticonvulsant obtained using a scanning electron microscope.

Figure 2. An electron micrograph of the described anticonvulsant obtained using a transmission electron microscope.

Figure 3. The particle size distribution of the described anticonvulsant in an aqueous suspension according to the DLS method.

Figure 4. IR spectra of mixtures of nanodiamonds with glycine used to construct calibration curves. I, II, III - spectra of mixtures with a glycine content of 1: 1.75: 2.5, respectively. Within the framework, characteristic peaks are highlighted.

Figure 5. Calibration curves for each characteristic band of the IR spectrum of a mixture of nanodiamonds with glycine, a, b, c - calibration curves for bands 1407, 1332 and 504 cm ^-1 , respectively.

The invention is illustrated by the following examples.

Example 1

300 mg of the initial detonation nanodiamond are annealed in a stream of hydrogen gas at a rate of 3.0 l / h at a temperature of 1000 ° C for 6 hours. Then, the annealed nanodiamond is subjected to liquid phase chlorination with molecular chlorine dissolved in 40 ml of CCl ₄ to 6 wt%. Cl ₂ . The chlorination reaction is carried out by photochemical exposure to visible light for 60 hours at a temperature of 60 ° C. The sample was then washed with CCl ₄ by centrifuging the suspension at 6000 rpm and dried under a pressure of 0.1 mm. Hg to constant weight. Then, a suspension is prepared from the chlorinated nanodiamond using 40 ml of a water-alcohol mixture (water: methanol = 1: 1), into which 300 mg of glycine as a free amino acid (NH ₂ CH ₂ COOH) is added with 1 ml of triethylamine. The resulting mixture was sonicated (50 W) for 60 minutes and kept under constant stirring at a temperature of 65 ° C for 30 hours. The resulting product was washed with a large amount of ethanol, centrifuged and dried under vacuum at 70 ° C overnight. The residual moisture content of the product is 2.2%. The yield of the target product is 279 mg (93%). The product is a dark gray with a bluish tint ultrafine powder (Figure 1) with primary particle sizes of 2-10 nm (Figure 2) having a shell of the surface layer up to 1 nm. In suspension, the particle size of the powder does not exceed 100 nm (Figure 3). The elemental composition of the particle surface of the obtained product is given in table.2.

table 2 XPS data of the obtained product Name of characteristic Chemical elements C O N Atomic% 80.1 ± 0.1 11.5 ± 0.1 8.4 ± 0.1 Communication energies, eV 285.2 ± 0.5 530.7 ± 0.5 399.6 ± 0.5

To determine the mass fraction of glycine in the resulting product, 3 mixtures of nanodiamond with glycine are prepared with a content of the latter 1: 1.75: 3.5, respectively. For each mixture, a weighed mass of 0.0035 g is taken and carefully ground in a mortar with 0.090 g of KBr. 0.070 g of the resulting mixture is pressed into a tablet and its IR spectrum is removed (Figure 4). Characteristic bands are selected at 1407, 1332 and 504 cm ^-1 , respectively, and calibration graphs are constructed for them (Figure 5). The intensity of the corresponding characteristic bands in the IR spectrum of the obtained sample with a mass of 0.0035 g was 0.23, 0.22, and 0.10 pu, respectively. From the calibration curves a, 6, c in FIG. 5, the glycine content in the obtained sample is determined, which is 0.00057 ± 8-10 ^-5 g. Therefore, the mass fraction of glycine in the sample sample is 21 ± 3% of the mass.

Example 2

The study of the specific anticonvulsant activity of the described agent.

The study was performed on white outbred adult mice males weighing 25-28 g. Animals were divided into 8 groups of 10 animals each, a total of 80 mice.

Animals were obtained from the Central laboratory animal nursery "Pillar", Moscow region. The keeping of animals corresponded to the rules of laboratory practice when conducting preclinical studies in the Russian Federation (GOST 3 51000.3-96 and 51000.4-96), the normative document "Sanitary rules for the design, equipment and maintenance of vivariums", approved by the Chief State Sanitary Doctor on 04/06/1973 No. 1045 -73, and Order of the Ministry of Health of the Russian Federation No. 267 of 06/19/2003 “On approval of laboratory practice rules” (GLP) in compliance with the International Recommendations of the European Convention for the Protection of Vertebrate Animals used in experimental studies (1997). The animals were kept in a vivarium at a temperature of 20-22 ° C, with a light cycle of 12 hours light and 12 hours dark periods, in T / 4A plastic cages measuring 580 × 375 × 200 mm with a stainless steel top lid and dust-free wooden bedding shavings. The animals were kept with constant access to feed and water using the full ration of extruded briquetted feed (GOST for feed P 50258-92) and drinking water. When conducting the experiments, the requirements of the Commission on the Ethics of Attitude to Animals of the Russian National Committee on Bioethics at the Russian Academy of Sciences and the ethical standards set forth in the "International Recommendations for Biomedical Research Using Animals" (1985) were taken into account. The experiments were carried out in the first half of the day.

The animal was subjected to maximum electroshock through special corneal (ophthalmic) electrodes by applying electrical stimulation (500/150 V / mA, 143-144 mA, 0.6 s). After the maximum electroshock, the animal developed tonic extension of the fore and hind limbs, ending in death. The ability of the described anticonvulsant to prevent the development of tonic extensia and to reduce the death of animals after conducting maximum electric shock was evaluated. Pharmacopoeial glycine was used as a comparison drug. The test substances were administered to animals once, intraperitoneally in a volume of 0.1 ml per 10 g of mouse weight 40 minutes before the maximum electroshock. The animals in the control group were injected with distilled water. Statistical processing of the results was carried out using statistical packages "BioStat" for Windows.

It was found that conducting maximal electroshock in control animals caused tonic extensia, ending in the death of 80% of mice, and only 20% of the animals survived after electroshock (Table 3). The described anticonvulsant at a dose of 10 mg / kg reliably protected 60% of mice from death, and at a dose of 15 mg / kg - 70% of mice after maximal electroshock (Table 3). Pharmacopoeial glycine in doses of 10 and 15 mg / kg did not affect seizures and did not eliminate the death of mice with maximum electroshock - all animals showed tonic extension and subsequent death (Table 3).

Table 3 Anticonvulsant activity of the described agent in the maximum electroshock test Substance, Dose, mg / ml % of animals without tonic expansion % surviving animals, ratio: (number of surviving mice / number of mice in the group) The control - 10% (1/10) 20% (2/10) Anticonvulsant 10 70% (7/10) * 60% (6/10) * Anticonvulsant fifteen 70% (7/10) * 70% (7/10) * Pharmacopoeial Glycine 10 0% (0/10) 0% (0/10) Pharmacopoeial Glycine fifteen 0% (0/10) 0% (0/10) * - p <0.05 significance of differences by the χ2 criterion relative to control

Thus, the anticonvulsant in doses of 10 and 15 mg / kg has a distinct anticonvulsant effect in the act electric shock test, statistically reliably preventing the development of tonic extension and death of animals.

To study the anticonvulsant activity of the described agent in the antagonism test, with chemical exposure, corazole was used, which was injected subcutaneously into the cervical region of the back. Due to the high hygroscopicity of corazole and the instability of the current dose, the dose-response curve for corazole was determined at the first stage of the study. Animals were observed for 60 min after injection of corazole with the registration of generalized seizures with loss of the reflex of inversion and death of animals. On the basis of these data, the dose causing convulsions in 97% of animals was calculated by probit analysis, which amounted to 105 mg / kg (ED ₉₇ ). Evaluation of the anticonvulsant effects of the substances according to the test of antagonism with corazole was carried out according to the indicators: the latent period of the appearance of the first generalized clonic seizures, the life expectancy of animals after administration of corazole and the percentage of death of animals.

The described anticonvulsant and pharmacopeia glycine were administered once intraperitoneally in a volume of 0.1 ml per 10 g of mouse weight 40 minutes before the administration of corazole. The animals in the control group were injected with distilled water. Statistical processing of the results was carried out using statistical packages "BioStat" for Windows. Group averages and standard error indicators were calculated.

It was established that in control animals, corazole caused first generalized clonic convulsions, then tonic convulsions and the death of animals. Convulsive manifestations and death of 100% of the control animals were observed after the administration of corazole at a dose of 105 mg / kg (Table 4). This dose was used to study the anticonvulsant effect of the described drug.

The described agent at a dose of 15 mg / kg caused a statistically significant increase (1.54 times) in the latent period of the appearance of the first generalized clonic seizures and an increase (2.64 times) in the life expectancy of animals compared to the control (Table 4). Along with this, the described tool significantly reduced (2 times) the number of dead animals (table 4). Pharmacopoeial glycine at a dose of 15 mg / kg did not exert anticonvulsant effect in the antagonism test with corazole (Table 4).

The data obtained indicate the presence of pronounced anticonvulsant activity in the test of antagonism with corazole of the described drug in a dose of 15 mg / kg (intraperitoneally, once) and the absence of an anticonvulsant effect in pharmacopeia glycine in a dose of 15 mg / kg.

Table 4 The study of the anticonvulsant activity of the described drug on the model of seizures caused by corazole Substance, dose The latent period of the first convulsions, min Life span, min Death,% The control 2.0 ± 0.15 14.41 ± 2.96 100% (10/10) Anticonvulsant, 15 mg / kg 3.08 ± 0.39 * 38.09 ± 4.62 * 50% (5/10) # Pharmacopoeial glycine, 15 mg / kg 2.2 ± 0.11 17.23 ± 3.16 90% (9/10) # - p <0.05 significance of differences by the χ2 criterion relative to the control; * - p <0.05 significance of differences according to the Mann-Whitney criterion compared with the control group.

Example 3

The study of side effects and toxic effects of the described anticonvulsant.

The study was conducted on white mongrel sexually mature male mice weighing 20-24 g, 2-3 months old. The experimental animals were kept as in Example 2. A total of 42 animals were used — 7 groups of 6 mice each.

Changes in behavior, reflexes, possible side effects, signs of intoxication, and death of animals were recorded after 1, 4, 24 hours, 4, 10, and 14 days. after intraperitoneal administration of the described anticonvulsant in comparison with the pharmacopoeial substance of glycine in the same doses.

As a result of the study, it was found that the described anticonvulsant when administered intraperitoneally to mice at doses of 75, 150 and 225 mg / kg did not cause signs of intoxication and death of the animals for 14 days. (tab. 5-7). At the same time, the described anticonvulsant also did not cause a change in the coat or mucous membranes in mice. There was also no ptosis of the upper eyelid, increased urination, defecation, salivation, piloerection, vocalization, lateral position. Within the normal range, there was a rhythm and depth of respiratory movements, there was no aggressiveness, timidity, tremor, convulsions, catalepsy, stereotype and grooming. No change in posture was observed. The animals retained pineal, corneal and pain reflexes. For all 14 days. observations, the animals were kept on an inverted mesh platform for 5 s (Table 5-7).

Table 5 The study of possible side effects and death of mice 1 hour after the administration of pharmacopoeial glycine and the described anticonvulsant (indicator of change in the group in%) Indicators Groups The control Anticonvulsant / dose, mg / kg Pharmacopoeial glycine / dose, mg / kg 75 150 225 75 150 225 Changing the nature of the coat 0 0 0 0 0 0 0 Change in the condition of the mucous membranes 0 0 0 0 0 0 0 Ptosis of the upper eyelid 0 0 0 0 0 0 0 Increased urination 0 0 0 0 0 0 0 Increased bowel movements 0 0 0 0 0 0 0 Increased salivation 0 0 0 0 0 0 0 Presence of piloerection 0 0 0 0 0 0 0 The presence of vocalization 0 0 0 0 0 0 0 Lateral position 0 0 0 0 0 0 0 Violation of the rhythm and depth of respiratory movements 0 0 0 0 0 0 0 Aggressiveness 0 0 0 0 0 0 0 Change in response to tapping a cell 0 0 0 33.3 * 0 0 33.3 * Increased shyness 0 0 0 0 0 0 0 Tremor 0 0 0 0 0 0 0 Cramps 0 0 0 0 0 0 0 Change pain thresholds 0 66.7 * one hundred* one hundred* 0 one hundred* one hundred*

Indicators Groups The control Anticonvulsant / dose, mg / kg Pharmacopoeial glycine / dose, mg / kg 75 150 225 75 150 225 reactions Posture change, catalepsy 0 0 0 0 0 0 0 Lack of coordination in the test of a rotating rod 0 16.7 fifty* fifty* 33.3 * 33.3 * fifty* 5 s holding on an inverted mesh platform one hundred one hundred one hundred one hundred one hundred one hundred one hundred Climbing from an inverted mesh platform one hundred 66.7 * 66.7 * fifty* 66.7 * 66.7 * 66.7 * The presence of a pineal reflex one hundred one hundred one hundred one hundred one hundred one hundred one hundred The presence of a corneal reflex one hundred one hundred one hundred one hundred one hundred one hundred one hundred Sedation 0 66.7 * one hundred* one hundred* 0 one hundred* one hundred* The presence of stereotype 0 0 0 0 0 0 0 The presence of grooming 0 0 0 0 0 0 0 Animal death 0 0 0 0 0 0 0 * - P <0.05 - relative to the control by the criterion χ2

Table 6 Study of possible side effects and death of mice 24 hours after the administration of pharmacopoeial glycine and the described anticonvulsant (indicator of change in the group in%) Indicators Groups The control Anticonvulsant / dose, mg / kg Pharmacopoeial glycine / dose, mg / kg 75 150 225 75 150 225 Changing the nature of the coat 0 0 0 0 0 0 0 Change in the condition of the mucous membranes 0 0 0 0 0 0 0 Ptosis of the upper eyelid 0 0 0 0 0 0 0 Increased urination 0 0 0 0 0 0 0 Increased bowel movements 0 0 0 0 0 0 0 Increased salivation 0 0 0 0 0 0 0 Presence of piloerection 0 0 0 0 0 0 0 The presence of vocalization 0 0 0 0 0 0 0 Lateral position 0 0 0 0 0 0 0 Violation of the rhythm and depth of respiratory movements 0 0 0 0 0 0 0 Aggressiveness 0 0 0 0 0 0 0

Indicators Groups The control Anticonvulsant / dose, mg / kg Pharmacopoeial glycine / dose, mg / kg 75 150 225 75 150 225 Change in response to tapping a cell 0 0 0 0 0 0 0 Increased shyness 0 0 0 0 0 0 0 Tremor 0 0 0 0 0 0 0 Cramps 0 0 0 0 0 0 0 Change in pain response thresholds 0 0 0 0 0 0 0 Posture change, catalepsy 0 0 0 0 0 0 0 Lack of coordination in the test of a rotating rod 0 0 0 0 33.3 * 16.7 33.3 * 5 s holding on an inverted mesh platform one hundred one hundred one hundred one hundred one hundred one hundred one hundred Climbing from an inverted mesh platform one hundred one hundred one hundred one hundred one hundred one hundred one hundred The presence of a pineal reflex one hundred one hundred one hundred one hundred one hundred one hundred 66.6 * The presence of a corneal reflex one hundred one hundred one hundred one hundred one hundred one hundred one hundred Sedation 0 0 0 0 0 0 0 The presence of stereotype 0 0 0 0 0 0 0 The presence of grooming 0 0 0 0 0 0 0 Animal death 0 0 0 0 0 0 0 * - P <0.05 - relative to the control by the criterion χ2

Table 7 The study of possible side effects and death of mice after 14 days. after the administration of pharmacopeia glycine and the described drug (indicator of change in the group in%) Indicators Groups The control Anticonvulsant / dose, mg / kg Pharmacopoeial glycine / dose, mg / kg 75 150 225 75 150 225 Changing the nature of the coat 0 0 0 0 0 0 0 Change in the condition of the mucous membranes 0 0 0 0 0 0 0 Ptosis of the upper eyelid 0 0 0 0 0 0 0 Increased urination 0 0 0 0 0 0 0 Increased bowel movements 0 0 0 0 0 0 0 Increased salivation 0 0 0 0 0 0 0

Indicators Groups The control Anticonvulsant / dose, mg / kg Pharmacopoeial glycine / dose, mg / kg 75 150 225 75 150 225 Presence of piloerection 0 0 0 0 0 0 0 The presence of vocalization 0 0 0 0 0 0 0 Lateral position 0 0 0 0 0 0 0 Violation of the rhythm and depth of respiratory movements 0 0 0 0 0 0 0 Aggressiveness 0 0 0 0 0 0 0 Change in response to tapping a cell 0 0 0 0 0 0 0 Increased shyness 0 0 0 0 0 0 0 Tremor 0 0 0 0 0 0 0 Cramps 0 0 0 0 0 0 0 Change in pain response thresholds 0 0 0 0 0 0 0 Posture change, catalepsy 0 0 0 0 0 0 0 Lack of coordination in the test of a rotating rod 0 0 0 0 0 0 0 5 s holding on an inverted mesh platform one hundred one hundred one hundred one hundred one hundred one hundred one hundred Climbing from an inverted mesh platform one hundred one hundred one hundred one hundred one hundred one hundred 66.7 * The presence of a pineal reflex one hundred one hundred one hundred one hundred one hundred one hundred one hundred The presence of a corneal reflex one hundred one hundred one hundred one hundred one hundred one hundred one hundred Sedation 0 0 0 0 0 0 0 The presence of stereotype 0 0 0 0 0 0 0 The presence of grooming 0 0 0 0 0 0 0 Animal death 0 0 0 0 0 0 0 * - P <0.05 - relative to the control by the criterion χ2

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Claims (4)

1. An anticonvulsant, which is glycine, immobilized on particles of detonation nanodiamonds with a size of 2-10 nm, having a shell thickness of up to 1 nm, with a glycine content of up to (21 ± 3) wt.%. 2. The method of obtaining an anticonvulsant according to claim 1, characterized in that the detonation nanodiamond is annealed in a stream of hydrogen gas at a temperature of 500-1200 ° C for 1-8 hours, subjected to liquid-phase chlorination with molecular chlorine by photochemical exposure to visible light at a temperature of 50- 70 ° C for 36-60 hours, followed by washing with carbon tetrachloride, centrifugation and drying, the resulting chlorine-modified nanodiamond is dissolved in a polar solvent to form a suspension, a tertiary amine is added and glycine, the resulting mixture is sonicated, followed by aging at 50-80 ° C, centrifugation, washing with a solvent and drying. 3. The method according to claim 2, where the ultrasound treatment is carried out for 5-60 minutes and keeping at 50-80 ° C is carried out for 12 to 48 hours 4. The method according to claim 2 or 3, where triethylamine is used as a tertiary amine and pyridine, a lower aliphatic alcohol, a water-alcohol mixture or water are used as a polar solvent.

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