RU2438691C1 - Medication possessing haemostimulating, antimutagenic, antitumour, cerebroprotective, antihypoxic, nootropic, anxiolytic and antineurotic action - Google Patents

Abstract

FIELD: medicine, pharmaceutics. ^ SUBSTANCE: invention relates to pharmaceutical industry, in particular to medication, which has haemostimulating, antimutagenic, antitumour, cerebroprotective, nootropic, anxiolytic and antineurotic action. Medication, which has haemostimulating, antimutagenic, antitumour, cerebroprotective, nootropic, anxiolytic and antineurotic action, represents a product of extraction of hairy root culture of roots of skullcap Baikal Scutellaria baicalensis Georg 70% with ethanol at defined conditions. ^ EFFECT: medication has increased haemostimulating, antimutagenic, antitumour, cerebroprotective, nootropic, anxiolytic and antineurotic action. ^ 11 tbl, 5 ex

Description

The invention relates to medicine, specifically to pharmacology.

The high frequency of occurrence and insufficient effectiveness of the treatment of diseases of the blood system and hematological complications, including iatrogenic in nature, associated with antitumor cytostatic therapy, as well as pathological conditions of the central nervous system, including disorders of higher nervous activity, is the basis for the development of new drugs possessing hemostimulating, antimutagenic, antitumor and psychotropic effects.

There are a large number of drugs with hemostatic, antimutagenic, antitumor, cerebroprotective, antihypoxic, nootropic, anxiolytic and anti-neurotic effects [1].

The closest in nature of origin and technical result to the proposed tool is an extract of the root of a natural plant of Scutellaria baicalensis Scutellaria baicalensis Georg [2]. This tool is selected as a prototype.

The disadvantage of this tool is its lack of hemostimulating, antitumor, cerebroprotective, antihypoxic, nootropic, anxiolytic and anti-neurotic activity and its lack of antimutagenic effect.

There is a technology for in vitro cultivation of the tissue of the roots of Scutellaria baicalensis Scutellaria baicalensis Georg, which provides a large mass of plant materials using a bioreactor for biotechnological cultivation of plant explants (Bioplant LLC, Tomsk).

We first revealed pronounced hemostimulating, antimutagenic, antitumor, cerebroprotective, antihypoxic, nootropic, anxiolytic and anti-neurotic activities of Scutellaria baicalensis Georg.

The problem solved by this invention is the expansion of the arsenal of drugs with hemostimulating, antimutagenic, antitumor, cerebroprotective, antihypoxic, nootropic, anxiolytic and anti-neurotic effects, and increasing the effectiveness of the treatment of tumor diseases and pathological conditions characterized by impaired hematopoiesis and disorders of the central nervous system.

The problem is achieved by using as an hemostatic, antimutagenic, antitumor, cerebroprotective, antihypoxic, nootropic, anxiolytic and anti-neurotic drug culture extract of Scutellaria baicalensis Georg, Scutellaria baicalensis root, which is a product of hairy root extraction (“hairy roots” in 70% culture 1) , 5 hours at a ratio of raw materials: extractant 1: 20 at a temperature of 80-85 ° C, followed by evaporation and drying to a moisture content of 5%.

New in the present invention is the use of an extract of tissue culture of the roots of Scutellaria baicalensis Scutellaria baicalensis Georg as a hemostimulating, antimutagenic, antitumor, cerebroprotective, antihypoxic, nootropic, anxiolytic and anti-neurotic agent.

The original tool we used - an extract of tissue culture of the roots of Scutellaria baicalensis, was developed and obtained by the Scientific Research Institute of Pharmacology SB RAMS (Tomsk) together with the Institute of Plant Physiology named after K.A. Timiryazev (Moscow).

The toxicity of antitumor drugs used in modern oncology significantly limits their practical application. Almost all drugs of this group have a pronounced damaging effect on the hematopoietic tissue and cause, to one extent or another, dysfunctions of the central nervous system and higher nervous activity, which are determined to a large extent by damage to the genetic apparatus of various cells of the body [3, 4]. In addition, the constant increase in the frequency of occurrence of diseases of the mental sphere, including the consequences of injuries and hypoxic damage to the central nervous system, and the insufficient effectiveness of their therapy with existing drugs [1] indicate the urgency of developing new psychotropic drugs.

It is known that the root extract of the natural plant of Scutellaria baicalensis Scutellaria baicalensis Georg has hemostimulating, antitumor, antimetastatic, anxiolytic, cerebroprotective action and some types of psychotropic activity [2]. At the same time, this tool does not have an antimutagenic effect, which, of course, affects the severity of its therapeutic properties. In addition, there is the difficulty of obtaining a homogeneous extract composition, associated with significant differences in the content of biologically active substances in natural plants, determined by climatogeographic and other environmental factors [5]. In contrast, the technology of tissue cultivation in vitro allows you to get the most homogeneous cultures, including those with given characteristics, by creating the same external conditions (temperature, composition of the nutrient medium, etc.) [6].

Nevertheless, information about the possibility of obtaining a pharmacologically more active extract of Scutellaria baicalensis Georg and the appearance of antimutagenic properties as a result of using the root culture of this plant as a substrate does not exist.

The fact of using the extract of tissue culture of the roots of Scutellaria baicalensis with the achievement of a new technical result, which consists in the effective stimulation of hematopoiesis, as well as antitumor, cerebroprotective, antihypoxic, nootropic, anxiolytic and anti-neurotic effects against the background of the appearance of anti-mutagenic properties, is not obvious for a specialist.

The experiment showed unpredictable results.

The claimed essential features in the aggregate showed new properties that are not explicitly derived from the prior art in this field. The present invention can be used in experimental medicine with access to practical health care. An identical set of features was not found in the study of the state of the art in patent and medical literature.

Based on the foregoing, the claimed technical solution should be considered relevant criteria: "Novelty", "Inventive step", "Industrial applicability".

The experiments were performed on 234 outbred mice and 672 linear mice (CBA / CaLac and C57BL / 6 lines), as well as Drosophila melanogaster flies. The mice were obtained from the nursery of the experimental biomedical modeling department of the Research Institute of Pharmacology SB RAMS (certificate is available).

The study of the pharmacological properties of the drug was carried out in accordance with the "Guidelines for the experimental (preclinical) study of new pharmacological substances" [7].

An extract of the roots of the natural Scutellaria baicalensis (EPSB) was used as a control.

Example 1

The roots of Scutellaria baicalensis were cultured - (hairy root culture) - under sterile conditions in 300 ml conical flasks containing 200 ml of liquid nutrient medium, with regular root explant transplants every 6 weeks. A device for the biotechnological growth of plant explants was a system of: a sealed chamber, a reservoir for a nutrient medium, inlet and outlet pipelines, a valve that regulates the supply of a nutrient medium, a spray device, filters for sterilization of incoming and outgoing gases, a pump for supplying a nutrient medium, a valve ( Bioplant LLC, Tomsk). The grown roots were lyophilized, dried to a moisture content of 7-8% and extracted with 70% ethanol in a water bath under reflux for 1-3 hours at a ratio of raw materials: extractant 1:20 at a temperature of 80-85 ° C. The resulting extract was evaporated on a rotary evaporator to a thick syrupy liquid and then dried in a thermostat at a temperature of 55 ° C to a moisture content of 5%. The output of the extract from raw materials is 20%, the content of the sum of flavonoids in terms of Baikalin is 30-35%.

Example 2

The hematopoiesis stimulation efficiency was studied in CBA / CaLac mice using a model of cytostatic myelosuppression induced by the introduction of MTD (maximum tolerated dose) of cyclophosphamide (250 mg / kg) in 0.3 ml of physiological saline. On the background of modeling myelosuppression, experimental animals were orally administered once a day for 9 days at a dose of 50 mg / kg with a culture extract of Scutellaria baicalensis (CSCM). Control animals in a similar mode received the extract of the roots of natural Scutellaria baicalensis ("Scutellaria baicalensis extract", GNTsLS, Kharkov, Ukraine). The dose used and the regimen for administering the agents in preliminary experiments were determined to be the most effective for the extract of Scutellaria baicalensis.

Blood system indices were determined using standard and cultural research methods on days 3, 5, and 10 after modeling myelosuppression [8].

The introduction of cyclophosphamide naturally led to the development of severe myelosuppression. There was a sharp drop in the content of cellular elements in the bone marrow and peripheral blood (Tables 1, 2).

The introduction of both drugs was accompanied by a significant acceleration of the regeneration of hematopoietic tissue. However, the most significant shifts were observed in the group of mice that received the extract of the culture of the Baikal shlemnika. So, the number of immature and mature neutrophilic granulocytes, as well as erythroid cells in the bone marrow in this case was 2.8 (5th day); 1.9 (5th day) and 3.7 (10th day) times higher than those in the control group, respectively. Moreover, these changes were a reflection of the reaction from the pool of the ancestral cells of granulomonocytic (CFU-GM) and erythroid (CFU-E) hematopoiesis cells (Table 3).

Thus, the culture extract of the roots of Scutellaria baicalensis tissue had pronounced hemostimulating properties, determined by the effect on the regulation system of hematopoiesis [9]. Moreover, the effectiveness of this tool significantly exceeds that of the standard extract of Scutellaria baicalensis.

Example 3

The study of antimutagenic properties.

The experiments were performed on CBA / CaLac mice and Drosophila melanogaster flies.

To evaluate the antimutagenic properties of Scutellaria baicalensis extract, the method of accounting for chromosomal aberrations in mouse bone marrow cells and the somatic mosaicism system for Drosophila melanogaster, recommended for such studies, were used [7].

Damage to the genome was carried out by a single and course (5 times after 48 hours) intraperitoneal injection: 1) cyclophosphamide (CF) at a dose of 20 mg / kg, or 2) cisplatin (CP) at a dose of 6 mg / kg, or 3) paclitaxel (PT ) at a dose of 40 mg / kg.

In experimental animals in the first series of experiments, a culture extract of the roots of Scutellaria baicalensis at a dose of 50 mg / kg was administered once to male mice. In the second series, the test agent at a dose of 50 mg / kg was administered to males and females for 5 days. Bone marrow cells were taken for study after a single injection 24 and 48 hours later, and only 24 hours after the administration of the cytostatic and corrector. As a control, groups of mice fed the extract of the roots of natural Scutellaria baicalensis (“Scutellaria baicalensis extract”, GNCLS, Kharkov, Ukraine) and universal antimutagen α-tocopherol with a course (10 times) intragastric administration at a dose of 1 ml / kg [ 7].

During the experiment, 1.5 hours before the end of the exposure (24 hours), 0.025% colchicine, 0.01 ml per 1 g of animal weight was injected intraperitoneally to accumulate metaphases. The bone marrow from the femur was prepared according to Ford and Hamerton / 11, 12 /. In all groups, 50-100 metaphases were analyzed from each animal. Undisturbed round-shaped cells with a good spread of chromosomes, without overlap, with a modular number of 40 were selected for analysis. The percentage of damaged cells in the bone marrow cells and the aberrant chromosomes per 100 cells were taken into account. Among structural chromosome abnormalities, single, paired fragments and exchanges were noted. Cells with multiple chromosome breaks were noted.

In the experimental groups of animals 24 hours after a single intraperitoneal administration of cytostatics, a significant increase in the level of aberrant chromosomes was observed - 72.53 ± 5.05% (of cyclophosphamide at a dose of 20 mg / kg), 37.00 ± 2.61% (of cisplatin in dose of 6 mg / kg), 7.23 ± 1.68% (of paclitaxel at a dose of 40 mg / kg) (Table 4).

With the introduction of experimental animals an hour before the introduction of cytostatics ECSB, a significant decrease in the level of aberrations was observed. So, the value of this indicator after 24 hours was 34.75 ± 4.17% (in combination with cyclophosphamide), 15.60 ± 0.51% (in combination with cisplatin) and 1.78 ± 0.48% (in combination with paclitaxel). Antimutagen Efficiency Factor (PEA) was 0.52, respectively; 0.57 and 0.75. Similar shifts occurred after 48 hours (Table 4).

EKSB also had a pronounced protective effect during its course administration (Table 5).

When studying the cytogenetic activity in EPSB and α-tocopherol, a complete absence of the antimutagenic effect was found in the first case and a significantly less pronounced effect in the second (Table 6).

In addition, the antimutagenic effect of the culture extract of Scutellaria baicalensis was confirmed using the method of somatic mosaicism on Drosophila melanogaster using yellow and singed markers [7].

During the experiment, antitumor drugs caused a significant increase in the number of females with mutations, while the addition of ECB to the nutrient medium at a concentration of 0.1% (which corresponds to a dose of 50 mg / kg for mice [7]) abolished the development of spontaneous mutations (χ ² = 17.77 compared to cyclophosphamide and χ ² = 6.26 compared to cisplatin). The PEA was respectively 0.61, 0.41 (Table 7).

In general, the results of the study indicate a pronounced gene protective effect (antimutagen) of the extract of the culture of the roots of Scutellaria baicalensis.

Example 4

In accordance with the requirements of the Guidelines for the Experimental (Preclinical) Study of New Pharmacological Substances [7], C57BL / 6 female mice with transplantable Lewis lung carcinoma studied the effect of ECB on the growth of the main tumor node and the development of metastases without cytostatic therapy and in the background intraperitoneal administration of cyclophosphamide at a dose of 125 mg / kg (on the 10th day after tumor inoculation).

An experimental animal was orally administered once a day for 9 days at a dose of 50 mg / kg with an extract of the culture of the roots of Scutellaria baicalensis (EKSM). Control animals in a similar mode received the extract of the roots of natural Scutellaria baicalensis ("Scutellaria baicalensis extract", GNTsLS, Kharkov, Ukraine). The dose used and the regimen for administering the agents in preliminary experiments were determined to be the most effective for the extract of Scutellaria baicalensis.

Evaluation of the effectiveness of therapeutic effects, which was carried out at the end of the course of treatment on the 21st day after tumor inoculation, showed that the isolated administration of C57BL / 6 female mice of both types of Scutellaria baicalensis did not lead to a decrease in the mass of the primary tumor (Table 8).

At the same time, the use of ECB and EPSB reduced the degree of dissemination of Lewis lung carcinoma, and in the case of ECB, there was also a decrease in the area of metastatic lung damage.

The proposed tool significantly increased the effectiveness of cytostatic therapy (table 8). In this case, not only the potentiating effect of the drug with respect to the antimetastatic effect of cyclophosphamide was observed, but also a significant increase in the inhibition of the growth of the primary tumor.

In particular, there was a decrease in the frequency of metastasis from 54% (only CF) to 33% and 20% with the additional introduction of the natural plant extract and its culture, respectively. In this case, corresponding changes in the number and area of metastases per 1 mouse were observed. In addition, inhibition of tumor growth was observed from 25% (CF) to 38% with the combined use of cytostatic with ECB, while EPS did not affect this indicator (Table 8).

Thus, the introduction of ECBB had a inhibitory effect both on the growth of the tumor node and on the process of dissemination and inhibition of hematogenous metastasis.

Example 5

Study of cerebroprotective and psychotropic activities.

The effect of ECSB on the neuropsychiatric status of mice that suffered hypoxic trauma, animals with posthypoxic encephalopathy, and its effect on behavior in conflict situations were investigated.

When studying cerebroprotective properties against hypoxia, animals were divided into 4 groups. During the experiment, part of the animals died during the modeling of hypoxia (these animals were not taken into account in the evaluation of the results). Some animals were rejected in the process of developing a conditioned passive avoidance reflex (passive avoidance reaction). Hypoxic effect was carried out on a hermetic volume hypoxia model [10]. The pressure chamber was a glass vessel with a calibrated volume of 500 ml (± <1.0%) and a hermetically sealed lid. The animals were placed in a pressure chamber, after which its lid was tightly closed. The mice were there until the death of a convulsive seizure or the first agonal inhalation, after which they were removed and allowed to "catch their breath." To obtain pronounced encephalopathy, hypoxic exposure was modeled twice with an interval of 48 hours. In this case, when using a 500 ml pressure chamber in animals, immediately after a hypoxic injury, a quick (within 2-3 hours) restoration of all functions is observed. Functional disorders from the side of the central nervous system caused by hypoxic exposure were assessed by the change in the orientation and research behavior in the open field and the safety of passive avoidance reaction.

With this formulation of the technique, within 10-15 days after the last hypoxia of the mouse, the mice exposed to it practically do not differ from normal animals, however, from 14-21 days on, mnemonic activity begins to appear in them, and by the end of the fourth week there is a partial death. Assessment of the condition of the animals by the safety of passive avoidance reaction was carried out 48 hours later, at 7, 14, 21, and 28 days after the development of the reflex. The death of animals was taken into account by the 35th day of the experiment. The results were evaluated by the proportion of animals with a preserved reflex and by the proportion of animals that died by the end of the fifth week after a hypoxic injury. Approximate research behavior in the open field and the production of passive avoidance reaction was carried out 1 hour after repeated hypoxic injury.

The “open field” experimental setup was a 40 × 40 × 20 cm camera with a square floor and white walls [11]. Its floor, divided into 16 squares, had a circular hole in each square with a diameter of 3 cm. From above, the camera was illuminated by an electric incandescent lamp with a power of 100 watts, located at a height of 1 m from the floor. The mouse was placed in one of its corners and for 3 minutes, separately in the first and two subsequent minutes, the number of movements from square to square (horizontal activity), the number of hind legs rising (vertical activity), and the number of hole examinations (mink) were recorded reflex), the number of washings (grooming) and the number of bowel movements according to the number of fecal balls (boluses), and the coefficient of asymmetry of behavior was calculated in the form of the ratio of the number of horizontal movements to total motor activity and expressed as a percentage. The results of the first and two subsequent minutes of testing were evaluated separately and in total.

The passive avoidance reaction method is based on the suppression of the innate reflex of preference for the dark space available in rodents [11]. The experimental setup was a camera consisting of two compartments: large - lighted and small - dark. The animal was placed in the light compartment and soon (after 10-20 seconds), due to the innate reflex of preference for dark space, it passed into the small compartment, after which the door connecting the two compartments was closed and fed to the floor of the dark compartment, consisting of parallel alternating electrodes, electric current pulses with a duration of 50 MS, a frequency of 5 Hz and an amplitude of 50 mA. After 10 seconds, the door was opened and the animal could jump out into the bright compartment with the usual floor. As a result of the described procedure, a conditioned reflex of avoiding dark space was developed in animals. When checking the reproducibility of the reflex, the animals were placed in the bright compartment at the angle opposite from the entrance to the dark compartment, and observed for 3 minutes. The time of the first entry into the dark compartment (latent time of entry) and the total time spent in the dark compartment were recorded. A reflex was considered developed if, during all 3 minutes of observation, the animal had never visited the dark compartment or the latent time of approach exceeded 150 s. The quality of the reflex was judged by the proportion of animals with the presence of a reflex. Additional indicators characterizing conditioned reflex activity and behavioral status were the number of bowel movements, grooming, the number of examinations of the entrance to the dark compartment, the time spent in the light compartment, the number of visits to the dark compartment and the time spent in the dark compartment.

Animals that, after being placed in the light compartment, remained motionless, and did not approach the entrance to the dark compartment, were not taken into account when calculating the results.

On the days of checking the safety of the reflex, animals were given EPSB ("Scutellaria baicalensis extract", State Scientific Center for Pharmaceutical Medicine, Kharkov, Ukraine) and the claimed drug in order to exclude the influence of the phenomenon of dissociation, at a dose of 50 mg / kg.

The Vogel conflict model is one of the most common neurosis models. It allows with sufficient reliability to predict the antieurotic properties of drugs [12]. The conflict situation was modeled by the collision of two reflexes: the drinking conditioned reflex and the unconditioned reflex of avoiding electric pain irritation. For this, the animals were previously left without water for two days, then for 4 days for 20 minutes they were daily placed in a plastic chamber 30 × 40 × 20 cm in size, in one of the end walls of which a niche with a drinking bowl was arranged, where the rat could get drunk . As a positive control (a drug whose performance indicators under the conditions of this technique characterize the validity of the results of this experimental study, a drug with known anxiolytic properties - phenazepam was used. Prototype (EPSB) at a dose of 50 mg / kg, the claimed drug in the same dose and a positive drug control - phenazepam in a dose of 1 mg / kg - was introduced, starting from the first day of training in the stomach 1 hour before placement in the chamber as a suspension in purified water, the control animals were injected with water throughout On the day of recording the results, 1 hour after the next injection of substances, the rats were again placed in the chamber and a potential difference was created between the metal floor of the chamber and the electrode lowered into the drinker so that when an attempt was made to take water, the animal received an electric shock. so that it has a suprathreshold value, increasing it during the first test testing on three test animals until the first reactions to avoid pain when the tongue touches the water. The number of water intake was recorded automatically by the number of circuit closures at the moment the tongue touched the water. The number of approaches to the drinking bowl was also recorded automatically using a photocell. The anxiolytic activity of the drugs was judged by the difference in the number of water intake, despite the shock (punished water intake), in the control and experimental groups. Additional indicators characterizing conditioned reflex, emotional and biased activity were the number of approaches to the drinking bowl, horizontal movements along the cage, grooming, and the number of vertical stands. Each animal was observed individually for 20 minutes.

Administration of drugs was started 4 days before hypoxic exposure and 5th time 1 hour before hypoxia, animals of intact and hypoxic controls received purified water. The indicated administration scheme allows us to evaluate the cerebroprotective effect, antihypoxic and nootropic effects of drugs of pharmacological substances [7].

The studies showed (Tables 9, 10) that the transferred hypoxia caused a decrease in the orientational research behavior in the open field with a shift in the coefficient of asymmetry of behavior from 54% to 72%. In the hypoxic control group, an increase in the latent time of entry into the dark chamber during the development of the reflex was noted, which indicates a violation of the orientational reflex. The use of the prototype led to a partial normalization of the orientation and research behavior in the open field, however, it was significantly inferior in this quality to the claimed tool. This is especially true of the indicative reflex, the severity of which when applying EPSB (prototype) tended to deteriorate even more, while the claimed tool had a normalizing effect on it. Manifestations of posthypoxic encephalopathy in animals of hypoxic control began from 14 days, progressively increasing by 28 days, while in this group the death of 65% of animals was noted. The use of both types of extract had a protective effect on animals, increasing the reproducibility of the reflex and reducing the death of animals, which indicated their cerebroprotective, antihypoxic and nootropic activities [7]. Moreover, the severity of effects when using the extract of the culture of the roots of Scutellaria baicalensis significantly exceeded that when using the prototype (table. 9, 10)

When studying the effect of drugs in conflict situations, phenazepam was found to have a pronounced anxiolytic effect for all the main indicators, which indicated the adequacy of the experimental conditions (Table 11) [7]. The prototype also showed anxiolytic properties, slightly inferior in activity to phenazepam in the main indicator of the method - the number of punishable water takes. The inventive tool found a pronounced anxiolytic effect, statistically significantly superior in activity to both the prototype and the positive control (the latter as a tendency), and indicating in this case also an anti-neurotic effect [7, 12].

The research results indicate the ability of ECSB to protect the central nervous system from the most common traumatic effects - hypoxia and psycho-emotional overstrain, and to correct disorders of higher nervous activity (cognitive functions) caused by pathological factors. Thus, the proposed tool exhibits cerebroprotective, antihypoxic, nootropic (antiamnestic) and anxiolytic properties.

In general, based on the data obtained, it follows that the extract of the culture of the roots of Scutellaria baicalensis has a pronounced hemostimulating, antimutagenic, antitumor, cerebroprotective, antihypoxic, nootropic, anxiolytic and anti-neurotic activities. Moreover, the breadth of the spectrum of therapeutic action of the proposed tool and its effectiveness are significantly superior to those of the extract of the roots of a natural plant.

Cited literature

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3. Voronova O. L., Pashinsky V. G. Pharmacological correction of the cytogenetic effect of cyclophosphamide. // Questions of oncology. - 1991. - N 2. - S.233-235.

4. Zhurkov B.C., Novakova Y.N., Scar R.Ya. The frequency of chromosomal aberrations in the bone marrow cells of mice when exposed to cyclophosphamide. Hygiene and sanitation, 1978, No. 1, pp. 12-14.

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Table 1 Peripheral blood indices in CBA / CaLac mice after administration of distilled water (1), 50 mg / kg of extract of natural Scutellaria baicalensis (2) and 50 mg / kg of extract of culture of Scutellaria baicalensis (3) against the background of simulation of cytostatic myelosuppression, (X ± m ) Terms of research
vania, day Red blood cells, T / L Reticulocytes, in ppm Band neutrophils, g / l Segmented neutrophils, g / l Platelets, T / L Background 8.61 ± 0.18 14.1 ± 0.3 0.40 ± 0.06 2.26 ± 0.02 796.57 ± 77.49 3rd one 7.89 ± 0.28 * 10.7 ± 0.24 * 0.01 ± 0.01 * 0.12 ± 0.03 * 503.80 ± 25.97 * 2 7.19 ± 0.21 * 9.89 ± 0.37 * 0.01 ± 0.01 * 0.13 ± 0.03 * 557.83 ± 46.69 * 3 7.84 ± 0.25 * 12.8 ± 0.6 * # 0.02 ± 0.01 * 0.17 ± 0.02 * 505.67 ± 24.25 * 5th one 7.53 ± 0.15 * 18.4 ± 1.3 * 0.16 ± 0.04 * 0.75 ± 0.23 * 349.40 ± 36.84 * 2 7.85 ± 0.17 * 27.6 ± 2.4 * # 0.13 ± 0.05 * 1.65 ± 0.10 * # 393.33 ± 33.96 * 3 8.59 ± 0.12 # $ 37.5 ± 1.89 * # $ 0.41 ± 0.08 # $ 1.7 ± 0.16 # 321.60 ± 23.22 * 10th one 7.29 ± 0.18 * 15.6 ± 1.4 1.31 ± 0.32 * 3.13 ± 0.23 * 977.33 ± 27.66 2 7.18 ± 0.15 * 16.7 ± 0.78 * 1.71 ± 0.42 * 5.36 ± 1.15 * # 843.33 ± 100.51 3 7.33 ± 0.15 * 16.4 ± 0.97 * 1.17 ± 0.23 * 7.42 ± 0.18 * # $ 884.17 ± 33.78 * - differences are significant in relation to the background,
# - differences are significant in relation to the group receiving distilled, with p <0.05
$ - differences are significant with the group of animals that received the extract of natural Scutellaria baicalensis, at p <0.05

table 2 Dynamics of bone marrow hematopoiesis in CBA / CaLac mice (× 10 ⁶ / thigh) after administration of distilled water (1), 50 mg / kg of extract of natural Scutellaria baicalensis (2) and 50 mg / kg of extract of culture of Scutellaria baicalensis (3) against the background of modeling cytostatic myelosuppression, (X ± m) Terms of research
Niya, day Immature neutrophilic granulocytes Mature neutrophilic granulocytes Eosinophilic granulocytes Lymphoid cells Monocytes Erythroid cells background 1.10 ± 0.10 3.72 ± 0.31 0.47 ± 0.09 3.38 ± 0.30 0.48 ± 0.06 1.79 ± 0.12 3rd one 0.04 ± 0.01 * 0.11 ± 0.04 * 0 1.87 ± 0.15 * 0.55 ± 0.04 0.18 ± 0.05 * 2 0.05 ± 0.02 * 0.16 ± 0.03 * 0 1.57 ± 0.13 * 0.60 ± 0.11 0.23 ± 0.03 * # 3 0.13 ± 0.03 * # 0.15 ± 0.04 * 0 1.36 ± 0.10 * # 0.42 ± 0.09 0.31 ± 0.02 * # 5th one 2.22 ± 0.26 * 1.45 ± 0.26 * 0.30 ± 0.08 2.03 ± 0.21 * 0.86 ± 0.08 * 1.04 ± 0.16 2 4.82 ± 0.46 * # 1.69 ± 0.23 * 0.07 ± 0.07 * 2.66 ± 0.26 0.98 ± 0.20 * 1.77 ± 0.18 # 3 6.31 ± 0.52 * # $ 2.2 ± 0.11 * # $ 0.12 ± 0.08 2.28 ± 0.36 1.29 ± 0.15 * 1.98 ± 0.30 # 10th one 0.80 ± 0.18 4.65 ± 0.45 0.76 ± 0.45 1.82 ± 0.25 * 0.60 ± 0.09 0.35 ± 0.06 * 2 0.80 ± 0.11 6.57 ± 0.53 * # 0.51 ± 0.10 1.87 ± 0.17 * 0.66 ± 0.14 0.39 ± 0.03 * 3 1.29 ± 0.06 # $ 6.11 ± 0.57 * # 0.33 ± 0.10 2.39 ± 0.50 0.94 ± 0.21 1.29 ± 0.19 # $ * - differences are significant in relation to the background,
# - differences are significant in relation to the group receiving distilled, at p <0.05
$ - differences are significant with the group of animals that received the extract of natural Scutellaria baicalensis, at p <0.05

Table 3 Dynamics of the content of CFU-E and CFU-GM in the bone marrow of CBA / CaLac mice (for 10 ⁵ non-adherent myelokaryocytes) after administration of distilled water (1), 50 mg / kg of extract of natural Scutellaria baicalensis (2) and 50 mg / kg of culture extract Scutellaria baicalensis (3) on the background of modeling cytostatic myelosuppression, (X ± m) Study time, days CFU-E CFU-GM Background 6.9 ± 0.73 4.35 ± 0.38 3rd one 9.50 ± 0.50 * 8.38 ± 0.63 * 2 10.88 ± 0.81 * # 10.00 ± 0.87 * # 3 13.13 ± 0.43 * # $ 12.38 ± 0.82 * # 5th one 7.25 ± 0.70 6.63 ± 0.73 * 2 12.50 ± 0.53 * # 5.75 ± 0.75 3 18.13 ± 0.44 * # $ 10.88 ± 0.55 * # $ 10th one 6.38 ± 0.68 4.25 ± 0.56 2 5.75 ± 0.37 5.50 ± 0.42 3 7.88 ± 0.83 7.14 ± 0.70 * * - differences are significant in relation to the background,
# - differences are significant in relation to the group receiving distilled, at p <0.05
$ - differences are significant with the group of animals that received the extract of natural Scutellaria baicalensis, at p <0.05

Table 4 Evaluation of the cytogenetic activity of paclitaxel, cisplatin and cyclophosphamide and their combinations with Scutellaria baicalensis in a test for accounting for chromosome aberrations in bone marrow cells of CBA / CaLac mice, X ± m. Experiment Conditions Study Dates The number of aberrations, (%) Polyploid cells, tetroloids, (%) The proportion of damaged cells (%) Cyclophosphamide 24 h 72.53 ± 5.05 0.20 ± 0.20 34.36 ± 1.38 48 h 10.82 ± 2.27 0.20 ± 0.20 8.33 ± 1.45 DF + ESCB 24 h 34.75 ± 4.17 * 0.25 ± 0.25 18.00 ± 1.29 * 48 h 4.50 ± 1.19 * 0.00 ± 0.00 4.00 ± 0.91 * CPU 24 h 37.00 ± 2.61 0.00 ± 0.00 24.2 ± 1.1 48 h 12.2 ± 0.42 0.00 ± 0.00 10.9 ± 0.3 CPU + EXB 24 h 15.60 ± 0.51 * 0.00 ± 0.00 12.00 ± 1.00 * 48 h 4.00 ± 0.55 * 0.20 ± 0.20 4.00 ± 0.55 * PT 24 h 7.23 ± 1.68 5.83 ± 1.51 6.56 ± 1.50 48 h 4.66 ± 0.93 19.90 ± 2.50 3.65 ± 0.50 PT + EXB 24 h 1.78 ± 0.48 * 15.02 ± 0.82 * 1.78 ± 0.48 * 48 h 1.73 ± 0.23 * 21.83 ± 0.57 1.73 ± 0.23 * * - significance of differences between experiment and cytostatic at p <0.05

Table 5 Assessment of the cytogenetic activity of cisplatin and cyclophosphamide and their combinations with the course administration of Baikal Scutellaria after 24 hours in a test for accounting for chromosome aberrations in bone marrow cells of mice - males of the CBA / CaLac line, X ± m. Experiment Conditions The number of aberrations, (%) Polyploid cells, tetroloids, (%) The proportion of damaged cells (%) CF 72.53 ± 5.05 0.20 ± 0.20 34.36 ± 1.38 CF + EXB 40.20 ± 2.20 * 0.00 ± 0.00 18.00 ± 1.29 * CPU 37.00 ± 2.61 0.00 ± 0.00 24.20 ± 1.10 CPU + EXB 17.60 ± 3.03 * 0.00 ± 0.00 11.40 ± 1.69 * * - significance of differences between experiment and cytostatic at p <0.05

Table 6 Evaluation of the cytogenetic activity of Cyclophosphamide and its correction with α-tocopherol in the test for accounting for chromosomal aberrations in bone marrow cells of male CBA mice Experiment Conditions Study Dates The number of aberrations, (%) Polyploid cells, tetroloids, (%) The proportion of damaged cells (%) CF 24 h 80.6 ± 5.96 0.20 ± 0.20 32.0 ± 1.3 48 h 3.60 ± 1.70 0.00 ± 0.00 3.20 ± 1.50 CF + α-tocopherol 24 h 68.5 ± 6.61 0.00 ± 0.00 26.5 ± 1.9 * 48 h 1.16 ± 0.32 0.00 ± 0.00 1.16 ± 0.32 CF + EPSB 24 h 81.6 ± 4.20 0.00 ± 0.00 31.60 ± 1.20 48 h 3,50 ± 0,30 0.00 ± 0.00 3,50 ± 0,60 * - significance of differences between experience and control at p <0.05

Table 7 Accounting for somatic mosaicism on Drosophila melanogaster using yellow and singed markers Drug dose The number of viewed females The number of females with mutations χ ² compared with control χ ² compared with the cytost. Stains "sn ³ " Stains "y" Spots "y sn ³ " Total spots CF 1096 16 67 0 83 58.19 - CF + EXB 1299 fourteen 31 0 45 22.54 17.77 PT 1336 26 37 0 63 40.54 - PT + EXB 535 5 13 0 eighteen 19.23 3.43 CPU 1154 61 28 3 92 61.85 - CPU + EXB 1000 42 5 four one 36.25 6.26 Eksb 769 four 3 one 8 0.86 - Control (Na Cl) 937 one 3 0 four - -

Table 8 The effect of Scutellaria baicalensis extracts on the development of Lewis lung carcinoma in female C57BL / 6 mice Groups Tumor mass (X ± m), g Inhibition of tumor growth,% The frequency of metastasis,% The number of metastases per 1 mouse (X ± m) Area of metastases per 1 mouse (X ± m), mm ² The control 4.56 ± 0.18 - one hundred 13.00 ± 0.50 14.83 ± 2.57 Eksb 4.15 ± 0.22 - one hundred 8.50 ± 0.90 * 7.38 ± 1.27 * EPSB 4.41 ± 0.23 - one hundred 10.6 ± 0.95 * 14.32 ± 2.3 CF 3.42 ± 0, N * 25.0 54 4.81 ± 0.22 * 5.97 ± 0.64 * CF + EXB 2.81 ± 0.07 * # 38,4 twenty 2.3 ± 0.17 * # 2.56 ± 0.27 * # CF + EPSB 3.44 ± 0.15 24.6 33 4.0 ± 0.1 * # 5.2 ± 0.72 * * - the significance of the difference between the indicator and the control group is noted at p <0.05,
# - the significance of the difference between the groups of mice that received only CF and CF together with extracts was noted, at p <0.05,

Table 9

The effect of the drink in comparison with piracetam on the indicators of the orientation and research behavior of outbred male mice in the open field after suffering a hypoxic injury in three minutes of observation

Group
observation dose Total motor activity Horizontal activity Vertical activity Mink reflex Grooming Defecation Asymmetry coefficient Cont
intact role 68 ± 9 * 37 ± 5 * 4.0 ± 1.5 23.3 ± 3.1 1.2 ± 0.6 1.8 ± 0.3 * 54 ± 3 * Cont
the role of hypoc
sic 36 ± 4 26 ± 2 1.4 ± 0.4 6.9 ± 1.0 1.9 ± 0.5 0.2 ± 0.1 72 ± 2 Prototype 46 ± 6 29 ± 4 * 3.5 ± 1.0 * 12.3 ± 2.0 * 0.9 ± 0.3 0.1 ± 0.1 63.7 ± 3 * The inventive tool 60 ± 4 * 35 ± 4 * 4,5 ± 0,6 19.5 ± 2.0 * # 0.5 ± 0.2 0,0 ± 0,0 58 ± 3 * * - the differences are significant in comparison with the hypoxic control at p <0.05

Table 10

The effect of nitrous oxide solution in comparison with piracetam on the safety of the conditioned reflex of passive avoidance of outbred male mice after a hypoxic injury

Observation groups, dose Hypoxia time, sec Latent time of entry into the dark chamber during reflex development, sec The proportion of animals with a preserved reflex when checked through (in%) The proportion of animals that died by the end of 35 days, in% 48 hours 7 days 14 days 21 days 28 days In-line control - 12 ± 2 * one hundred* one hundred* one hundred* 92 one hundred* 0 * Hypoxic control 57.2 ± 4.1 49 ± 2 83 83 33 17 17 65 Prototype 61.3 ± 5.6 68 ± 7 one hundred* one hundred* 83 * 67 * 46 twenty* The inventive tool 75.9 ± 4.8 * # 32 ± 4 * # one hundred* one hundred one hundred* 92 * # 92 * # 0 * # * - the differences are significant in comparison with the hypoxic control at p <0.05;
# - the differences are significant in relation to the group receiving the prototype, with p <0.05

Table 11

The impact of the proposed drug in comparison with the prototype and the positive control (phenazepam) on the behavioral indicators in rats in a conflict situation in arbitrary units (conflict)

Indicators Observation group The control Phenazepam 1 mg / kg Prototype (EPSB) Hairy Root Tissue Culture Extract (EXB) Horizontal activity 45 ± 14 61 ± 9 78 ± 20 59 ± 13 Vertical activity 17.7 ± 2.7 14.3 ± 0.4 30.3 ± 7.6 17.7 ± 4.5 Grooming 16.56 ± 3 1.9 ± 0.6 15.6 ± 5.5 16.8 ± 4.4 The number of boluses (fecal balls) 0 ± 0 0 ± 0 0 ± 0 0 ± 0 Fade time 393 ± 32 134 ± 23 * 100 ± 21 * 88 ± 26 * Time spent at the drinking bowl, sec 96 ± 66 204 ± 46 84 ± 18 218 ± 42 * # Number of drinking approaches 4.6 ± 1.3 13.5 ± 2.6 6.4 ± 1.3 4.8 ± 0.9 Number of Water Takes 30 ± 4 71 ± 7 * 58 ± 7 92 ± 8 * # Drinking time, sec 38 ± 3 52 ± 5 * 100 ± 13 187 ± 27 * # Total motor activity 79 ± 16 91 ± 6 123 ± 31 93 ± 26 * - differences are significant in comparison with the control at p <0.05;
# - the differences are significant in relation to the group of animals that received the prototype at p <0.05.

Claims (1)

An agent with hemostimulating, antimutagenic, antitumor, cerebroprotective, antihypoxic, nootropic, anxiolytic and anti-neurotic effects, characterized in that it is a product of hairy root extraction of the Scutellaria baicalensis Georg Scutellaria baicalens roots roots 70% with ethanol for 1 to 3 hours: extractant 1:20 at a temperature of 80-85 ° C, followed by evaporation and drying to a moisture content of 5%.