RU2279877C2 - Adaptogen - Google Patents

Abstract

FIELD: organic chemistry, medicine, biochemistry.

SUBSTANCE: invention proposes using acizol representing bis-(1-vinylimidazol-N) zinc diacetate as an adaptogen. Acizol is known early as antidote. Acizol exceeds plant Eleutherecoccus senticosus by activity with respect to indices of functional state of the body adaptation systems due to providing the higher intensity of tissue respiration, improvement of energetic metabolism and regenerative processes.

EFFECT: enhanced effectiveness and valuable medicinal and biochemical properties of adaptogen.

1 dwg, 8 tbl

Description

The present invention relates to medicine, in particular to a new adaptogen.

Adaptogens include herbal preparations (native or in the form of pure active substances) that have a low-specific effect on the central nervous system functions, endocrine regulation, metabolic processes and increase the body's adaptation to adverse conditions [1].

The mechanism of adaptogenic effects (including glycerides of Eleutherococcus) is due to the weakening of negative biochemical and functional changes during stress reactions and the activation of adaptive synthesis of RNA and proteins, leading to improved energy metabolism and recovery processes. For the development of a noticeable effect, a long time and regular intake are required (peak after 4-6 weeks with daily intake).

The most widely used and affordable is the extract of Eleutherococcus [2].

The disadvantage of this known method is the relatively low efficiency and the need for a long and regular intake.

The objective of the invention is a new adaptogenic agent with higher efficiency.

This problem is solved by using acizole as an adaptogen.

It is known to use acizole, bis (1-vinylimidazole-N) zinc diacetate, as an antidote for carbon monoxide poisoning [3].

The choice of directions for studying the pharmacological activity of the drug “Atsizol” was determined by the alleged indications for its use, the main of which is the adaptation of the body to adverse and extreme environmental factors (ionizing radiation and activation of free radical reactions, oxidative damage, hypoxia, energy-deficient states). The same considerations guided the choice of comparison drugs.

From comparative load tests, we used models of antagonism with hexenal (anti-narcotic effect) by the duration of hexenal sleep [4], taking into account the duration of restoration of the ability to rectilinear movement after rotation according to K. G. Vasiliev [5], the maximum duration of static work (keeping your body with white mice on a vertical grid), the duration of swimming mice with cargo (dynamic work) [6]. Of the models of extreme factors, we used modeling of hypoxia in a closed volume ("can hypoxia") [7].

Acyzole and a reference drug were administered daily per os for two weeks to white non-linear mice using an atraumatic-tip dispenser at 10 a.m. Doses of the drugs were extrapolated from humans to rodents and amounted to: Atsizol - 10 mg / kg, Eleutherococcus - 1 ml / kg.

The dynamics of the weight of animals was determined on a VLR-500 scale.

The content of reduced glutathione in the liver was determined iodometrically [8], sulfhydryl groups of the blood serum were determined by amperometric titration according to N. S. Rubina [9], and the glycogen content was determined by the Samody method [10].

Blood for biochemical studies was obtained after decapitation of animals.

The content of total protein and lipids, cholesterol, bilirubin of blood serum was determined using the Bio-La-Test kits of the Czech company "Lahema". The glucose level in the blood and organ homogenates was determined by the ortotoluidine method.

The intensity of tissue respiration in organ homogenates was determined by the Warburg manometric method [11, 12]. The content of ATP is chromatographic [13].

Statistical processing of the experimental results was performed according to Student-Fisher.

Hexenal antagonism

The antagonism of drugs with hexenal was determined after two weeks of their daily oral administration to mice. The control group of animals received placebo - distilled water in the same volume. Hexenal was administered subcutaneously at a dose of 70 mg / kg. The test results are presented in table 1.

The data obtained indicate that Atsizol has a significant anti-narcotic effect (the duration of hexenal sleep decreased), that is, the drug activates the detoxifying monooxygenase system of the liver.

Recovery of ability to rectilinear motion after rotation

A test of accounting for the duration of restoration of the ability to rectilinear movement after rotation demonstrates the effectiveness of the drug to increase the adaptation of the vestibular drug to overload. Radial acceleration was created by rotating mice in a centrifuge for 15 seconds at a speed of 600 rpm. The mice were placed in cylindrical plastic tubes, closed on the outside with a wire mesh. Usually, after centrifugation, the mice extracted from the centrifuge rotated around the longitudinal axis of the body or made maneuvering movements (moving in a circle).

In the experiment, groups of animals were used after daily oral administration for 14 days of the study drug and the reference drug. Each group consisted of 10 males. At the same time, an equal number of animals from each group were rotated. The results are presented in table 2. They indicate that Atsizol significantly reduced the duration of the recovery period after rotation, i.e. improved the ability of the vestibular apparatus to adapt to radial acceleration.

The effect of drugs on the static-mechanical endurance of mice

The effect of the drugs on the static-force endurance was studied by recording the hanging time of mice of the experimental and control groups on a vertical grid. Animals were tested after two weeks of daily drug administration. The criterion for the depletion of static strength was considered the time when the mouse could no longer support the weight of its body and fell down from the net (the dead weight of the mice averaged 28 g). The experimental results are presented in table 3. They demonstrate an increase in static physical endurance under the influence of all drugs.

The effect on the duration of swimming mice

Swimming is a heavy physical dynamic load, allowing to evaluate the adaptogens efficiency [14]. It was carried out with a load (a lead tube on a rubber ring attached to the root of the tail), equal to 5% of body weight, at a water temperature of 38-39 ° C. The criterion of fatigue and cessation of swimming was considered the first "diving" with the immersion of the nasal passages in water. In a large bath, 5 animals from each observed group were simultaneously swimming. Testing of mice was performed 14 days after daily administration of drugs (table 4).

These tables show that the drugs increase the duration of swimming, and Atsizol is not inferior to the registered analogue.

Antihypoxic effect

Respiration from a confined space - re-respiration - is a fairly adequate and simple model of acute hypoxia [15, 16]. The animal, absorbing oxygen from an enclosed space due to breathing, causes the development of its deficiency - hypoxic hypoxia, which makes it possible to evaluate the studied drug by the integrated mortality rates for a certain observation time and resistance to oxygen deficiency (maximum life expectancy).

The animals were placed in a 250 ml jar tightly closed with a glass lid lubricated with sealant. The maximum lifespan and symptoms of thanatogenesis were recorded using a stopwatch. Cans with animals during the study were in the air conditioner, ensuring the constancy of the experimental conditions (temperature + 20 ° С, humidity - 65-70%, atmospheric pressure - 760 ± 10 mm Hg, st). The controls were intact animals.

After death, the brain was obtained ex tempore in each animal, its hemogenesis was performed (400 rpm, 10 strokes of the pestle) [17], and the catalase activity, the level of diene conjugates and lipid hydroperoxides were determined in the homogenate [18, 19]. These indicators allow you to evaluate the antioxidant properties of the drug.

It turned out that the drugs significantly increase the life time of mice, while lipid peroxidation rates stabilized (levels of malondialdehyde and hydroperoxides of brain lipids decreased) and antioxidant (catalase) activity was restored, which indicates an increase in reserve antioxidant activity of the brain (table 5).

Indicators of the functional state of the body's adaptive systems

The indicators characterizing energy metabolism, lipid metabolism and antitoxic activity of the body are presented in table 6.

The presented results indicate the adaptive focus of the studied drugs.

Immobilization stress

Immobilization stress was modeled on male rats after 20 days of daily (including weekend) administration of the drug at a dose of 10 mg / kg. For this, rats were fixed on their back for 24 hours in the boxes of the Golubev chamber. The development of the adaptation syndrome was judged by body weight, weight of the thymus, adrenal glands, spleen and the number of hemorrhages (ulcers) in the gastric mucosa [20]. The controls were intact animals. The results are presented in table 7.

It turned out that after daily immobilization in animals treated with placebo (distilled water), body weight significantly and significantly decreased, the adrenal gland mass increased by 30%, the thymus gland mass decreased by 20%, and the spleen mass decreased by 25%. In the stomach, ulcerative lesions and hemorrhages developed. This testified to the development of the adaptation syndrome, i.e. stress. Both drugs prevented a decrease in body weight, changes in the weight of the adrenal glands, thymus gland and spleen. The number of ulcerative lesions was on average 80% less. This allows us to talk about the anti-stress activity of the drug "Atsizol" with its prophylactic purpose. The positive effect on the course of the stress reaction once again confirms the adaptogenic orientation of the pharmacological activity of the drug Atsizol.

The effect of Atsizol on physical performance and resistance to physical exertion

A study of the influence of Atsizol on physical performance, endurance and resistance of the body to prolonged intense loads was carried out with a single and course use of the drug. The state of severe physical fatigue was achieved by crossing a cross-country terrain (10 km), followed by overcoming an obstacle course of 100 m (including 50 m with a simulated “victim” weighing 70 kg).

The first study was conducted with the participation of 31 healthy men aged 18 to 23 years, 17 people in the experimental group and 14 in the control placebo group. Acizole at a dose of 120 mg (2 ml of a 6% solution) and placebo were administered orally by the double-blind method immediately before exercise.

Immediately after the cross, direct performance indicators (static and dynamic components and carpal muscle endurance), the functional state of the cardiovascular system, and well-being were examined. When studying direct performance indicators in the control group after exercise, a significant decrease in the dynamic performance indicator (up to 68 ± 5%, p <0.05) and a sharp (up to 23 ± 5%, p <0.05) drop in the static endurance indicator were revealed (Table 8). The endurance coefficient according to wrist dynamometry decreased from 0.79 ± 0.03 to 0.55 ± 0.02 units. (p <0.01).

The state of the cardiovascular system was characterized by fatigue of the myocardium (Kvaas index increased by 40% relative to the background). The activity of the sympathoadrenal system increased (heart rate increased by 121%, Robinson index by 80%). A significant decrease in the tolerance of the cardiovascular system to physical activity was revealed: the myocardial tolerance index and PWC ₁₇₀ decreased by 40%, the mechanical performance of the heart - by 27%, the specific maximum oxygen consumption - by 17%. IOC increased by the end of the load by 23% of the background value.

Thus, intense prolonged physical activity caused the development of fatigue, accompanied by a significant tension in the reserves of the cardiovascular system.

The state of the central nervous system was characterized by an increase in the subjective assessment of the degree of fatigue by almost 3 times (from 2.1 ± 0.2 to 6.9 ± 0.2 points on a 10-point scale), well-being and activity decreased by 12% and 9%, respectively (p <0.05.

Acizol intake had a significant impact on the indicators of dynamic and static endurance (an increase of 43% from the placebo level, p <0.01 and 13%, respectively, p <0.05). Acizole significantly increased the tolerance of the cardiovascular system to stress. He increased by 15% the values of the PWC ₁₇₀ test indicators and the productivity of mechanical work of the heart, by 18% - myocardial tolerance to loads (p <0.05). The stroke volume increased compared with the control by 18% with a significant decrease in the metabolic demand of the myocardium for oxygen (Robinson index decreased by 38%, p <0.01) and an increase in its functional reserves (Kvaas index decreased by 17%, p <0.05 ), which naturally led to a decrease in heart rate by 14% (p <0.01).

Such an effect of Atsizol on the studied hemodynamic indices and physiological indices allows us to explain the fact that the subjects from the experimental group went cross over time 12% faster (p <0.05) than the tested placebo groups. Moreover, this difference in running speed was most pronounced in the last third of the route, especially during overcoming an obstacle course.

The data obtained show that a single prophylactic use of Acyzole increases the stamina and resistance of the body to prolonged intense loads and improves their subjective tolerance.

In a second study, the effect of a 10-day course of acyzole (orally 120 mg once a day after meals) on physical endurance and body resistance to prolonged intense stress was studied. The study was conducted with the participation of 34 healthy men aged 18 to 23 years, 20 of them in the experimental and 14 in the placebo group. We investigated direct performance indicators (static and dynamic components and carpal muscle endurance), the functional state of the cardiovascular system, and a subjective assessment of well-being. The state of severe physical fatigue was achieved 4 days after the last dose of Atsizol and placebo, as in the previous study, by crossing a cross country (10 km) and then overcoming an obstacle course of 100 m (including 50 m with a simulator of the “injured” weighing 70 kg). The study was conducted immediately after the cross.

The results of the studies showed that after the course use of Acyzole, the indicators of dynamic and static endurance in the experimental group exceeded by 18 and 24% the similar indicators in the placebo group (Fig. 1).

Acizol intake had a positive effect on the parameters of carpal physical endurance (an increase of 23% from the placebo level, p <0.05). It was found that Atsizol increased by 16% the values of the PWC ₁₇₀ test indicators, the performance of mechanical work of the heart and myocardial tolerance to loads by 11%. A significant increase in its functional reserves was revealed (Kvaas index decreased by 17%), which naturally led to a decrease in heart rate by 14%. The Stange test was 17% higher compared with the placebo group.

These data indicate a positive effect of the course application of Atsizol on the tolerance of the cardiovascular system to stress.

In the experimental group, well-being, activity, mood were also noted, in addition, the drug significantly facilitated the tolerance of physical exertion, which was manifested in a decrease in the feeling of muscle fatigue, shortness of breath while running, a decrease in pain manifestations in the right hypochondrium, an even running pace was easier to maintain (Fig. 2 )

The subjective assessment of the feeling of fatigue was significantly lower than in the placebo group subjects.

The data obtained allow us to say that the positive effect of Atsizol on physical performance, endurance and resistance to prolonged intense exertion persists several days after the end of the course of the drug.

Thus, the above data indicate the high activity of Acyzole as an adaptogen and its advantages over the known pharmacological analogue.

Literature

1. Register of Medicinal Products of Russia. Encyclopedia of Medicines, 10th issue, 2003, p. 1063.

2. Ibid., P. 1064.

3. Patent No. 2038079 "Carbon monoxide antidote", IPC A 61 K 32/515, 1995

4. Kudrin A.N. Fiziol. zhur. USSR, 1953, vol. 39, No. 3, p. 309-318.

5. Vasiliev K.G. Gig. labor and prof. Sick., 1957, No. 2, pp. 19-24.

6. Brekhman I.I. Ginseng. L .: 1957, p. 30-33.

7. Guide to physiology. Human adaptation to extreme environmental conditions. Ed. O.G. Gazenko. M., "Science", 1979, S. 333-336.

8. Ellouk-Achard S. et al. Ex vivo and in vitro models in acetaminophen hepatotoxicity studies. Relationship between glutathione depletion, oxidative stress and disturbances in calcium homeostasis and energy metabolism. Archives of Toxicology. Supplement. 1995, 17, pp. 209-214.

9. Timer M., Gedrich I. Pharmacology and Toxicol., 1969, No. 5, p. 602-604.

10. Kigel G.B. Kharabajahyan A.V. Indicators of biological norms for laboratory animals. Rostov-on-Don, 1978, 95 pp.

11. Clinical evaluation of laboratory tests. Ed. N.U. Titsa, M., "Medicine", 1986, 480 p.

12. Umbrayt V.V., Burris R.Kh. et al. Monometric methods for studying tissue metabolism. M., 1957, 356 p.

13. Laboratory research methods in the clinic. Handbook Ed. V.V. Menshikov. M., "Medicine", 1987, p.141-143.

14. Rylova M.L. Research methods for the chronic effect of harmful environmental factors in the experiment. M .: "Medicine", 1964, 228 p.

15. Guide to physiology. Ecological physiology of man. Human adaptation to extreme environmental conditions. Ed. O.G. Gazenko. M., "Science", 1979, S. 333-336.

16. Guide to practical exercises by pathological physiologists. Ed. O.M. Pavlenko. M., "Medicine", 1974, p. 174-175.

17. Tyson CA, Luman KD, Stephens RJ Age-related differences in G-SH-shuttle enzymes in NO ₂ -or O ₃ -exposed rat lungs Arch. Oh _s . Env. Health, 1982, Vol. 37., No. 3, pp. 167-176.

18. Research methods in occupational pathology. Ed. O.G. Arkhipova. M., 1988, p. 156-158.

19. Modern methods in biochemistry. Ed. V.N.Orekhovich. M., "Medicine", 1977, S. 62-64.

20. Kigel G.B., Kharabadzhakhyan A.V. and other indicators of the biological norm for laboratory animals. Rostov-on-Don, 1978, p. 25-28.

Table 1
The duration of the hexenal sleep of mice after preliminary administration of drugs (M ± m) Groups of animals Sleep duration, min The control 24.8 ± 2.1 Acizole 15.5 ± 1.3 * Eleutherococcus 20.2 ± 2.0 * - significant differences from control at P <0.05 table 2
The duration of the restoration of the ability to rectilinear translational movement of mice after rotation in a centrifuge for 15 seconds at a speed of 600 rpm (M ± m) Groups of animals Recovery time, sec The control 210 ± 8 Acizole 145 ± 5 * Eleutherococcus 165 ± 7 * * - significant differences from control at P <0.05 Table 3
The physical endurance of mice in the test of static power load own weight (M ± m) Groups of animals Hanging time, min The control 22.0 ± 1.5 Acizole 28.5 ± 1.0 * Eleutherococcus 27.5 ± 1.0 * * - significant differences from control at P <0.05

Table 4
The duration of swimming mice with cargo after drug administration (M ± m) Groups of animals Duration of swimming, min The control 27.8 ± 5.5 Acizole 43.8 ± 6.6 * Eleutherococcus 40.2 ± 6.3 * * - significant differences from control at P <0.05 Table 5
The effectiveness of the drugs in the test "can hypoxia" (M ± m) Groups of animals Indicator Life span, min Malonic dialdehyde, nmol / mg protein Catalase, μmol H ₂ O ₂ / mg min Hydroperoxides of lipids, units opt. Densities at 480 im Intact - 2.77 ± 0.26 9.57 ± 0.58 0.11 ± 0.01 The control 34.2 ± 2.6 4.61 ± 0.55 3.50 ± 0.50 0.50 ± 0.04 Acizole 49.0 ± 1.0 * 3.19 ± 0.47 * 5.55 ± 0.50 * 0.25 ± 0.03 * Eleutherococcus 44.2 ± 1.0 * 3.47 ± 0.41 4.76 ± 0.58 * 0.40 ± 0.03 * - significant differences from control at P <0.05

Table 6
Indicators of the functional state of the body's adaptive systems Indicators Groups of animals The control Acizole Eleutherococcus Body weight g 27.0 ± 1.4 26.5 ± 1.3 27.2 ± 1.1 Total lipids, serum, g / l 3.6 ± 0.2 3.5 ± 0.1 2.9 ± 0.1 * Cholesterol, serum, mmol / l 1.64 ± 0.32 1.59 ± 0.33 1.60 ± 0.28 Bilirubin total, serum, mmol / l 3.1 ± 0.2 2.9 ± 0.2 3.4 ± 0.1 -SHGR, serum, mg% 1634 ± 83 1680 ± 80 1686 ± 79 Reconstituted glutathione, liver, mg% 165 ± 12 166 ± 10 153 ± 10 Glycogen, liver, mg% 2560 ± 110 2520 ± 115 2400 ± 115 Glucose, serum, mg% 92 ± 15 91 ± 18 83 ± 13 Glucose, muscle, mg% 135 ± 27 145 ± 37 155 ± 29 ATP, heart, µmol / g 2.5 ± 0.2 3.0 ± 0.2 * 2.3 ± 0.1 ATP, muscles, mmol / g 5.3 ± 0.2 5.9 ± 0.4 * 5.1 ± 0.3 Intensity of tissue respiration, heart, l O _2/100 mg / hour 63 ± 6 67 ± 6 60 ± 6 Intensity of tissue respiration, the muscles, l O _2/100 mg / hour 21 ± 3 28 ± 5 22 ± 2 * - significant differences from control at P <0.05

Table 7
Body mass, adrenal gland, thymus, spleen and the number of hemorrhages in the gastric mucosa in male rats in the immobilization stress test A drug No. of animals Body weight g M ± m Adrenal glands, mg M ± m Thymus mg M ± m Spleen, mg M ± m Hemorrhage M ± m one 2 3 four 5 6 7 8 9 10 eleven 12 Control (intact animals) one
2
3
four
5
6
7
8 160
170
165
175
180
155
150
160


164 ± 4 38
42
37
35
40
35
33
37


37 ± 1 13
fourteen
eleven
10
fifteen
eleven
10
9


12.0 ±
1.0 240
220
230
210
215
230
210
210


200 ± 5


-


0 Placebo (stress) one
2
3
four
5
6
7
8 140
135
150
150
145
155
150
147


146 ± 3

P _1,2 <
0.05 48
52
56
58
57
58
54
fifty


54 ± 2

P _1,2 <
0.05 9
8
9
7
8
5
7
8


8.0 ±
0.5

P _1,2 <
0.05 170
200
150
165
170
175
160
155


168 ± 6

P _1,2 <
0.05 12
10
8
fourteen
fifteen
10
9
eleven


11 ± 1

P _1,2 <
0.05

Continuation of table 7 one 2 3 four 5 6 7 8 9 10 eleven 12 Acizole 10 mg / kg (stress) one
2
3
four
5
6
7
8 165
170
150
150
155
170
165
150


159 ± 3

P _1.4 >
0.05
P _2.4 <
0.05 fifty
42
39
40
42
36
35
37


40 ± 2

P _1.4 >
0.05
P _2.4 <
0.05 12
fourteen
eleven
9
8
10
eleven
12


10.9 ± 0.7

P _1.4 >
0.05
R _2.4 >
0.05 170
190
200
180
210
230
210
215


201 ± 8

P _1.4 >
0.05
P _2.4 <
0.05 one
0
2
one
3
2
four
5


2 ± 1


P _2.4 <
0.05

Table 8
The effect of acizole on indicators of physical endurance, performance and hemodynamic parameters after prolonged exercise (% of background) Indicator Placebo N = 14 Acizole N = 17 % to placebo Static endurance 23 ± 5 * 33 ± 5 143 ** Dynamic stamina 68 ± 3 * 77 + 5 113 * Carpal endurance 68 ± 5 ** 79 ± 15 116 PWC ₁₇₀ 62 ± 5 * 73 ± 5 115 * The IOC 123 ± 8 * 124 ± 13 101 With 104 ± 5 123 ± 7 118 * IPC specific 83 ± 3 * 87 ± 5 104 Heart mechanical performance 73 ± 5 * 83 ± 3 114 * ITM 60 ± 3 ** 71 ± 3 * 118 * IR 140 ± 8 ** 116 ± 10 83 IR 180 + 22 * 112 ± 18 62 ** Sample Stange 69 ± 19 * 81 ± 15 115 * Note: The difference is statistically significant: * - p <0.05, ** - p <0.01

Claims (1)

The use of acizole as an adaptogen.

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