RU2496504C1 - Agent for extreme performance stimulation, method for preparing and using it - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine. The agent represents a mixture of peptides of molecular weight 500-900 Da prepared of the brain of cattle and/or pigs of one year old or younger, particularly the midbrain (mesencephalon), namely the central grey substance surrounding the Sylvian aqueduct (aquaeductus cerebri Sylvii) with the peptide mixture concentration in the agent of 5.0-10.0 mg/ml of the aqueous solution or the content of 5.0-10.0 mg/g in a witepsol suppository. The method involves grinding the cattle and/or porcine frozen midbrain, hydrolysis at +20°C in the presence of pepsin, extraction, separation of the extract from ballast substances by separation through materials with retention potential more than 2000 Da, separation of the purified extract by fluid extraction at critical temperature CO₂ 31.1°C and critical pressure 7.3±0.1 atm, separation of the fraction of 500-900 Da, dissolution in water and freeze drying. The agent is applied in the form of the solution in water in the active substance concentration of 5.0-10.0 mg/ml for single parenteral administration in a dose of 10-30 mg, or in the form of a witepsol suppository with the active substance concentration of 5.0-10.0 mg/ml for single rectal administration in a dose of 10-30 mg.

EFFECT: group of inventions provides higher performance efficiency and tolerance in trained individuals at least by 10-12% as compared with the reference group.

3 cl, 5 ex, 10 tbl, 2 dwg

Description

The invention relates to medicine and relates to medical devices for a specific effect on body functions.

Known technical solutions made at the level of inventions that increase the working capacity and endurance of persons subject to intense physical exertion [1-3].

The indicated technical solutions are a food product [1] enriched in phospholipids or protein-amino acid food products [2] or protein-peptide modules for food production [3] for nutrition of athletes or other persons leading an active lifestyle.

It is characteristic that the above food products or additives to food products, firstly, do not significantly increase working capacity and endurance, and only after prolonged (2-3 months or more) consumption of these products, and secondly, the increase in working capacity and stamina occurs mainly due to an increase in muscle mass and, thirdly, these foods are not stimulants of performance and endurance as such, i.e. do not meet the necessary requirements for work in extreme conditions.

In the available scientific, technical and patent literature, no analogues and prototype are found that meet the proposed stimulator for working in extreme conditions.

The technical result, the achievement of which the creation of this invention is aimed, is to increase the working capacity and endurance when working in extreme conditions (extreme and transcendental and prolonged physical exertion, stressful situations, etc.), for example, during special operations and similar critical events requiring maintaining high performance in conditions of extreme psycho-emotional stress.

Also the technical result, the achievement of which the creation of this invention is directed, is to obtain a highly active agent for stimulating work in extreme conditions.

The technical result is achieved by the fact that the claimed tool for stimulating work in extreme conditions is a mixture of peptides with a molecular weight of 500-900 Da, obtained from the brain of cattle or pigs under the age of one year, in particular from the midbrain (mesencephalon), namely, from the central gray matter surrounding the sylvian aqueduct (aquaeductus cerebri Sylvii) at a concentration of peptides in a medium of 5.0-10.0 mg / ml in an aqueous solution or 5.0-10.0 mg / g in a suppository.

The technical result is also achieved by the fact that the method of obtaining funds for stimulating work in extreme conditions includes the separation from the brain of cattle or pigs under the age of one year, the midbrain (mesencephalon), namely the central gray matter surrounding the sylvian aqueduct (aquaeductus cerebri Sylvii), freezing its grinding of frozen parts, homogenization, hydrolysis in the presence of pepsin, extraction and separation of the extract from ballast substances by separation through materials with with a retention capacity of more than 2000 Da and sedimentation of the fraction up to 2000 Da with 30% acetic acid solution, washing the precipitate with acetone, dissolving the precipitate in water and isolating the fraction 500-900 Da from the dissolved extract by supercritical fluid extraction at a critical temperature of CO ₂ 31.1 ° C and critical a pressure of 7.3 ± 0.1 atm., dissolving the fraction of 500-900 Da in water and drying by lyophilization and then preparing the product by dissolving the fraction of 500-700 Da at a mass content of 5.0-10.0 mg / ml in an aqueous solution or by introducing a lyophilized fraction of 500-900 Yes in vitepsol to a content of 5.0-10.0 mg / g and the subsequent formation of suppositories.

As raw materials for producing peptides, parts of the brain departments of slaughtered cattle and / or pigs (cattle) no older than 1 year old, in particular the midbrain (mesencephalon), namely the central gray matter surrounding the sylvian aqueduct (aquaeductus cerebri Sylvii), are used.

The hydrolysis of the crushed parts of the brain is carried out either in an acidic medium (acetic acid + zinc chloride) or in the presence of proteolytic enzymes - serine proteases - fungal serine endoprotease, pepsin, pectophostidine (P10X), etc.

The separation of the active components of the agent during hydrolysis and separation of the hydrolyzate is carried out by ultrafiltration and preparative chromatography, and in particular by supercritical fluid extraction.

The proposed tool for stimulating work in extreme conditions is an aqueous solution of a mixture of peptides with a molecular weight of 500-900 Da obtained by supercritical fluid extraction of purified and filtered hydrolyzate of parts of the brain parts of slaughtered cattle and / or pigs (cattle) not older than 1 year , in particular the midbrain (mesencephalon), namely the central gray matter surrounding the sylvian aqueduct (aquaeductus cerebri Sylvii), with a concentration of 5.0-10.0 mg / ml or a content of 5.0-10.0 mg / g per suppository Rhee based on Witepsol.

It should also be noted that the claimed tool does not cause either addiction or addiction (addiction).

Another significant feature of the claimed tool is that it can be detected in the blood of the recipient by analytical means only during the first 15-25 minutes. after parenteral administration (and with rectal administration, it cannot be detected at all by modern analytical means).

This circumstance is crucial in the practice of stimulating athletes.

Get a tool to stimulate work in extreme conditions as follows.

In slaughtered animals (11-month-old calves and 8-month-old pigs), the brain is removed, the central gray matter of the midbrain surrounding the sylvian aqueduct (B) is excised, these parts of the brain are frozen at least to -40 ° C [4] and then crushed and homogenized in water, the homogenate is then hydrolyzed at + 20 ° C in the presence of pepsin, then the hydrolyzate is extracted, the extract is separated from the ballast substances by separation through materials with a retention capacity of more than 2000 Da, the purified extract of fluid ext by fraction at a critical temperature of CO _{2 of} 31.1 ° C and a critical pressure of 7.3 ± 0.1 atm., a fraction of 500-900 Da is separated, dissolved in water and dried by lyophilization; (yield 0.35 g / 100 g of raw material), the preparation is further prepared by dissolving the fraction of 500-900 Da in water to a concentration of 5.0-10.0 mg / ml or by introducing the fraction of 500-900 Da in vitepsol to a content of 5.0 -10.0 mg / g and subsequent molding of suppositories.

The study of the impact of the claimed funds on improving adaptogenic activity, performance and endurance of humans (and animals) in a state of high physical and psycho-emotional stress is carried out in accordance with GOST R 15.105-2001, according to standard operating procedures of laboratories and according to the "Rules of laboratory practice in the Russian Federation "(Order of the Ministry of Health of the Russian Federation No. 267 of 06/19/2003), as well as the national standard of the Russian Federation" Principles of Good Laboratory Practice tics ”GOST R 53434 - 2009.

Example 1

The change in physical performance of outbred white rats under aerobic power loads is determined.

The effect of the claimed means on physical performance is determined by measuring the duration of the run on the treadmill from the start of the run to the moment the animal develops a state of the final refusal to perform physical activity. The duration of one test cycle is 27 days.

The test is carried out as follows. Preliminary training of animals at the stand (treadmill, 1 day, 10 min at an angle of inclination of the treadmill tape 20 ° and its speed 23 m / min), the introduction of funds to animals (within 21 days), observation of animals after stopping the introduction of funds (7 days) . A repeated test cycle with the introduction of funds is carried out 30 days after the 21st day (termination of administration of the drug). The cycle is thus 21 days, 30 days, 21 days, etc.

Form 2 groups of laboratory rats of 12 animals each. The first group receives the claimed drug, the second - (control) - saline. Drugs are administered rectally. Doses of administration: agents 0.29 mg / goal., 40 minutes after administration of the agent - run 15 minutes at a speed of 27 m / min and with a tape inclination of 20 °. The voltage is 40 V. The indicators are taken on the 1st, 8th, 15th, 21st and 27th day of the experiment. The main indicator is the total contact time of the rat with the electric platform and the running time.

The endurance of rats when running on a treadmill is presented in Table 1.

Table 1 The inventive tool 1st day 8th day 15th day 21st day 27th day running (sec) 254 ± 25 258 ± 18 259 ± 28 229 ± 15 206 ± 11 Control group running (sec) 241 ± 26 203 ± 23 168 ± 21 133 ± 13 143 ± 13

As can be seen from Table 1, the endurance of rats compared with the control group increases significantly.

Figure 1 (see page 8 of the description) presents the effect of the claimed funds on the endurance of experimental animals from the original (100%). Areas of 20 days represent the use of the claimed drug, 30 days - a break in the application. On the abscissa axis, the test time (days) is postponed and on the ordinate axis, the change in endurance as a percentage of the initial value.

Figure 2 (see page 8 of the description) presents the effect of the claimed funds on the endurance of experimental animals in contact with an electrical stimulating platform treadmill. Test time (days) is plotted on the abscissa axis and contact duration is plotted on the ordinate axis. Lines 1 and 2 show the effect of the claimed drug and data for the control group, respectively.

Example 2

To identify the pharmacological activity of the claimed drug, a test of "despair" or forced swimming, which reflects the state of depression of animals. The forced swimming test is a combined hard form of stress that combines physical and emotional components.

In the experiment, animals of both groups are subjected to stress - swimming in a pool with a load. Water temperature + 24 ° C.

The test results are presented in Table 2

table 2 The inventive tool 1st day 8th day 15th day 22nd day 27th day First swim 79.7 ± 5.4 125.5 ± 6.2 178.0 ± 6.4 218.0 ± 6.6 197.8 ± 4.7 Repeated 58.2 ± 4.2 91.9 ± 5.3 144.0 ± 7.6 187.3 ± 5.4 162.0 ± 3.3 Third 66.8 ± 4.3 101.3 ± 5.5 147.8 ± 3.8 181.6 ± 6.7 176.3 ± 4.2 Control group First swim 63.8 ± 3.3 66.8 ± 2.6 71.0 ± 3.2 71.9 ± 3.9 91.4 ± 3.3 Repeated 49.3 ± 3.5 46.4 ± 2.4 52.7 ± 2.7 54.5 ± 3.0 68.7 ± 3.2 Third 53.9 ± 2.7 54.7 ± 2.2 62.3 ± 3.0 62.3 ± 3.4 79.8 ± 2.9

Example 3 (Hypoxia Test)

A measure of the sensitivity of animals to acute hypobaric hypoxia is the life expectancy at the "height". Each animal is "raised" to a critical height (11.5 thousand m) at a speed of 165 m / s, where it was located to an agonal state.

Designations:

Runway - the time of the first fall (transition to a lying position), characterizing the threshold of the body's reaction to this effect.

VZh - life time at "height" (before the appearance of agonal breathing).

GDP - the time of posture recovery after the animal “descends” from the “height”

KEZ - coefficient of effectiveness of protection (lifetime in experience / lifetime in control)

The test results of acute hypobaric hypoxia are presented in Table 3

Table 3 Indicator 1 day 8 day 15 day 22 day 27 day The inventive tool Runway, sec 10 ± 1,1 10.8 ± 1.0 9.2 ± 1.0 9.2 ± 1.2 10.4 ± 0.9 VZh, sec 104.2 ± 6.2 113.8 ± 5.4 128.8 ± 6.4 191.3 ± 5.9 186.3 ± 6.5 GDP, sec 24.6 ± 2.3 22.1 ± 2.1 17.9 ± 1.9 16.3 ± 1.5 17.1 ± 0.9 Control group Runway, sec 7.92 ± 0.74 8.75 ± 0.65 7.5 ± 0.75 8.75 ± 0.65 10 ± 0.62 VZh, sec 105.8 ± 6.2 92.9 ± 5.5 95.8 ± 3.7 107.1 ± 6.3 97.1 ± 5.1 GDP, sec 24.2 ± 2.6 21.3 ± 1.5 23.3 ± 2.3 25.8 ± 2.0 25.4 ± 1.9

Protection effectiveness indicators (CEC) * for acute hypobaric hypoxia (to its own background indicators) are presented in Table 4

Table 4 Groups KEZ 8 day 15 day 22 day 27 day The inventive tool 1.09 1.24 1.83 1,59 Control group 1,0 1,0 1,0 1,0

Example 4

The impact of the proposed drug on large laboratory animals (mini-pigs) at maximum physical exertion.

Form 2 groups of 6 heads of mini-pigs weighing 18 ± 1.9 kg, males, age 8 months. The first group receives the claimed drug, the second - (control) - placebo. Drugs are administered rectally. Dose of administration: 0.28 mg / kg, a rectal form of placebo was administered to the control group.

Animals are forced to run on a treadmill (Torneo T-203). The duration of the run and the distance covered are measured.

The endurance indicators of mini-pigs when running on a treadmill are presented in Table 6

Table 6 The inventive tool 1st day 8th day 15th day 21st day 27th day running time (min) 26.0 ± 1.1 28.1 ± 1.0 29.8 ± 0.6 31.0 ± 1.2 31.3 ± 1.4 distance traveled (m) 2600 ± 130 2900 ± 120 3100 ± 100 3300 ± 150 3400 ± 100 control group duration (min) 26.6 ± 1.2 27.0 ± 1.4 27.9 ± 1.3 28.2 ± 0.9 28.2 ± 1.9 distance traveled (m) 2500 ± 120 2600 ± 150 2700 ± 150 2900 ± 150 2900 ± 150

It should be noted that, along with an increase in endurance, mini-pigs also increased their speed of movement.

Example 5

Tests of the effects of the claimed funds are carried out on volunteer athletes.

Cyclists are selected (12 people with close anthropometric data, age 20-22), swimmers are also selected (12 people with close anthropometric data, age 18-20 years). Both cyclists and swimmers are divided into 2 groups of 6 people each. The first group receives the claimed drug, the second - (control) - saline. Drugs are administered rectally. Dose of administration: 10.0 mg.

The effect of the claimed means on the physical performance of cyclists is determined by measuring the duration of the “races” on the adjustable-load bicycle ergometer (911S) from the start of the “race” until the athlete develops a state of final refusal to perform physical activity. Table 7 presents the effect of the claimed funds on the physical performance of cyclists at maximum speed (spurt)

Table 7 The inventive tool 1st day 8th day 15th day 21st day 27th day the duration of the "race" (min) 4.2 ± 0.1 5.6 ± 0.2 6.1 ± 0.3 6.7 ± 0.1 6.6 ± 0.2 control group the duration of the "race" (min) 4.3 ± 0.3 4,5 ± 0,1 4.6 ± 0.2 4.6 ± 0.5 4,5 ± 0,1

Table 8 presents the effect of the claimed funds on the physical performance of cyclists at the stayer speed

Table 8 The inventive tool 1st day 8th day 15th day 21st day 27th day distance traveled (m) 7200 ± 130 8900 ± 120 10400 ± 120 11600 ± 100 10900 ± 100 control group distance traveled (m) 6300 ± 120 6500 ± 150 6900 ± 100 7100 ± 100 6900 ± 150

The influence of the claimed means on the overall physical endurance of swimmers (crawl) is determined by measuring the duration of swimming at a speed of 60-62% of the competition from the start of the swim until the athlete develops a state of refusal to perform physical activity. Table 9 shows the effect of the claimed drug on the overall physical endurance of swimmers.

Table 9 The inventive tool 1st day 8th day 15th day 21st day 27th day swimming time (min) 25.0 ± 1 27.0 ± 3 31.0 ± 2 35.0 ± 3 34.0 ± 3 control group swimming time (min) 24.0 ± 2 25.0 ± 3 24.0 ± 2 23.0 ± 5 22.0 ± 4

The effect of the proposed tool on the speed capabilities of swimmers is determined by measuring the distance traveled over 5 minutes of swimming at a speed of presumably ~ 95% of the competition. Table 10 presents the effect of the claimed funds on the speed capabilities of swimmers.

Table 10 The inventive tool 1st day 8th day 15th day 21st day 27th day Speed (km / h at 95% load) 4.85 ± 0.1 5.8 ± 0.1 6.2 ± 0.1 6.7 ± 0.1 6.5 ± 0.1 control group Speed (km / h at 95% load) 4.9 ± 0.1 5.0 ± 0.1 5.1 ± 0.1 5.0 ± 0.1 4.7 ± 0.1

As can be seen from the description, examples of the impact of the claimed drug on performance in extreme conditions (especially when exposed to extreme physical and psycho-emotional stress), as well as from comparison with control examples, the claimed tool stimulates an increase in performance and endurance in trained people by at least 10-12 % compared to the original,

Information sources

1. RU 2370158 C2, M.C. A23L 1/30, publ. 2010 year

2. RU 2375923 C2, M.C. A23L 1/30, publ. 2010 year

3. RU 2388350 C2, M.C. A23L 1/30, publ. 2010 year

Claims (3)

1. A tool to stimulate work in extreme conditions, characterized in that it is a mixture of peptides with a molecular weight of 500-900 Da, obtained from the brain of cattle or pigs under the age of one year, in particular from the midbrain (mesencephalon), namely, from the central gray matter surrounding the sylvian aqueduct (aquaeductus cerebri Sylvii), when the concentration of the mixture of peptides in the agent is 5.0-10.0 mg / ml in an aqueous solution or in the content of 5.0-10.0 mg / g in suppositories on Vitepsol based. 2. The method of obtaining funds for stimulating work under extreme conditions according to claim 1, characterized in that the brain is removed from slaughtered animals, the central gray matter of the midbrain (mesencephalon) surrounding the sylvian aqueduct (aquaeductus cerebri Sylvii), these parts of the brain are excised frozen at least to -40 ° C, then crushed and homogenized in water, the homogenate is hydrolyzed at 20 ° C in the presence of pepsin, then the hydrolyzate is extracted, the extract is separated from the ballast substances by separation through material s with retention capacity of over 2000 Da, separated purified extract fluid extraction under the critical temperature of CO ₂ 31,1 ° C and critical pressure (7,3 ± 0,1) atm., separated fraction 500-900 Yes, it is dissolved in water and dried by lyophilization and then prepare the tool by dissolving the obtained fraction in water at a concentration of 5.0-10.0 mg / ml or by introducing the obtained fraction into vitepsol to a content of 5.0-10.0 mg / g, followed by molding suppositories. 3. The use of funds to stimulate work in extreme conditions according to claims 1 and 2 in the form of an aqueous solution with an active substance concentration of 5.0-10.0 mg / ml for a single parenteral administration in a dose of 10-30 mg or in the form of a suppository based Vitepsol with an active substance content of 5.0-10.0 mg / g for a single rectal administration in a dose of 10-30 mg.

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