RU2276991C1 - Tonic and actoprotector remedy - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: remedy comprises mixture of phytoecdysteroids of 20-hydroxyecdysone and 25S-inocosteron extracted from aerial plant top of Serratula genus of Asteraceae family, preferably from leaves gathered in vegetation phase, mass budding phase or at the beginning of flowering.

EFFECT: enhanced effectiveness of treatment; wide range of medical applications.

Description

The invention relates to nutritional supplements and therapeutic drugs tonic and actoprotective effects and can be used as a new tool for asthenic and asthenodepressive conditions, accompanied by a weakening of the processes of protein synthesis, with prolonged infections and intoxications, with neurasthenia, neurosis and hypotension, as well as in athletes during intensive training for cardiovascular dysfunction, especially with severe signs of myocardial overstrain and increased protein catabolism in preparation for the competition.

Known tonic "Ekdisten" (A.S. 1312774 USSR, Tonic // Abubakirov N.K., Sultanov MB, Syrov V.N. et al .; Mamatkhanov A.U., Shamsutdinov M. - R .I., Shakirov TT The isolation of ecdysterone from the roots of Rhaponticum carthamoides // Chemistry of natural compounds. - 1980. - N4. - C528-530), the active principle of which is ecdysteroid 20-hydroxyecdysone obtained from roots with rhizomes of wild-growing or cultivated plants of the safflower levzea (Rhaponticum carthamoides (Willd.) Iljin, family Asteraceae).

The method of producing Ekdisten tonic from a plant of the genus Rhaponticum involves extraction of raw materials with 70% ethanol, removal of related substances from the concentrated extract by extraction with organic solvents, purification of the product by column adsorption chromatography on alumina and its subsequent crystallization. The product yield to the mass of raw materials is 0.1%.

The disadvantage of this analogue is the low percentage yield of ecdysteroids. In addition, the use of roots with rhizomes as raw materials reduces the reproducibility of medicinal plants, which indicates high economic costs for creating a raw material base.

The objective of the present invention is to expand the arsenal of tonic effects on a living organism, providing annually reproducible raw materials with a high content of ecdysteroids, obtaining a drug of low cost.

This is the technical result.

The technical result is achieved by using a mixture of ecdysteroids isolated from the aerial parts of plants of the genus Serratula of the Asteraceae family, as a tonic and actoprotective agent, preferably leaves collected during the vegetation phase, mass budding or the beginning of flowering.

A known mixture of ecdysteroids isolated by the method described in the patent of the Russian Federation No. 2153346, containing 20-hydroxyecdysone - at least 75% and 25S-inocosterone - at least 10% with the structural formulas:

which are active substances that exhibit physiological activity similar in magnitude to the analogue drug.

The proposed tool, called us "Serpisten", is a white crystalline powder, odorless, slightly soluble in water, highly soluble in methanol and 70% ethanol.

A mixture of phytoecdysteroids is obtained in a known manner (according to patent 2153346) as follows.

Harvesting of raw materials, preferably leaves, is carried out in the phase of vegetation, mass budding or the beginning of flowering. It was experimentally established that in these phases of development the content of active ecdysteroids is maximum. A study of the distribution of ecdysteroids in plants of the crown-sickle was carried out in middle-aged generative individuals. Crowned Serpukha far surpasses safflower rape by the content of ecdysteroids. The use of the aerial part of the plant, unlike the rhizomes of rapontica, greatly facilitates the harvesting and processing of ecdysteroid-containing plant materials.

The crushed plant material (the aerial part of the plant of the genus Serratula of the Asteraceae family) is repeatedly extracted with water at a temperature not exceeding 100 ° C for no more than 1 hour with constant stirring and the ratio of raw materials: extractant is 1:10, respectively. The thick aqueous extract was evaporated under reduced pressure, treated with a mixture of ethyl acetate-methanol taken in a ratio of 4: 1, or chloroform-methanol in a ratio of 6: 1. The procedure is repeated until complete extraction of ecdysteroids from the thick extract. The organic extracts are evaporated to dryness, then the ecdysteroids are purified by chromatography on alumina in a chloroform-methanol or ethyl acetate-methanol system of increasing polarity and recrystallization in an ethyl acetate-methanol or ethyl acetate-ethanol system in a 9: 1 ratio.

An example implementation of the method.

6 kg of crushed leaves of a plant of the genus Serratula of the Asteraceae family are poured into 60 l of water and extracted for 1 hour at a temperature of 50 ° C with constant stirring, then the extract is drained, and the plant material is subjected to re-extraction. The combined extract is evaporated in a vacuum rotary film evaporator under reduced pressure to 2 L. 2 l of the thick extract is treated three times with 8 l of a mixture of ethyl acetate-methanol in a ratio of 4: 1 with constant stirring and filtered. The filtrate was evaporated to dryness, the residues were combined and transferred to a column of aluminum oxide and eluted with a mixture of chloroform-methanol of increasing polarity. The eluate is collected in flasks of 1 l and determine the content of ecdysteroids in them by thin layer chromatography (TLC). The desired fractions containing ecdysteroids are evaporated to dryness and recrystallized in ethyl acetate-methanol (9: 1).

Thus, the method allows to obtain the substance "Serpisten", which is a mixture of phytoecdysteroids containing at least 75% 20-hydroxyecdysone and at least 10% inocosterone (white crystalline powder, odorless, slightly soluble in water (1.9 g / l) , it is soluble in 70% ethanol (61 g / l) and methanol (75 (g / l)), which is used as a tonic and actoprotective agent.The yield is 42 g, or 0.7%, to the mass of raw materials, which accounts for 70% of the theoretically calculated yield.

A known method for the isolation of 20-hydroxyecdysone and 25S-inocosterone (RF patent 2155599, which allows to obtain individual compounds from a mixture of phytoecdysteroids and mix them in an effective amount.

Verification of the tonic and actoprotective effect of the substance "Serpisten" was carried out in laboratory conditions on laboratory mice of two groups: white outbred and linear CBA. A total of 113 animals of three ages were studied: adults (6 months), young (2.5 months) and immature (1 month). The body weight of the animals was: white sexually mature males 25.3 ± 3.8 g, immature mice of both sexes 10.2 ± 1.3 g, linear males CBA 18.7 ± 2.6 g. Mice were kept in individual cells on a standard diet feeding in the same vivarium conditions (20 ° C), All animals were pre-screened for body weight, endurance (maintaining body weight on a pole) and swimming ability. The tested substance "Serpisten" in the form of an aqueous solution (at the rate of 0.2 ml per 20 g of body weight) was administered to mice per os in doses of 1 mg / kg and 10 mg / kg for 5 and 10 days. Control animals were kept under the same conditions at the same time and received per os distilled water (placebo). The mass of each animal was measured daily; accordingly, the injected volume of the solution was adjusted to it. A total of five groups of animals were studied: control (placebo) and four experimental groups that received per os Serpisten in final doses: 5 mg / kg, 10 mg / kg, 50 mg / kg, and 100 mg / kg. In all animals, control and experimental measurements of the parameters of strength and endurance under dynamic and static load were carried out, the reactivity of higher nervous activity was measured and the central nervous system tone was evaluated. Physical performance and the effect of the substance “Serpisten” on it was designated as the actoprotective effect of the substance “Serpisten” "and evaluated by standard methods (Elizarova, 1962). The endurance to dynamic load in animals was evaluated by the swimming time in the standard test (swimming to failure) when water temperature 30 ° C. Static load was measured by the maximum weight of the load that the animal can lift, and by the time of hanging on a pole.The tone of the central nervous system and the effect of the substance “Serpisten” on it started as the “central nervous system-tonic effect of the substance" Serpisten "" and evaluated by the speed of finding the animals out of the double T-shaped labyrinth (Tolmen, 1980). All measurements were carried out twice: on the first and fifth day of testing, which excluded the possibility of learning animals. Statistical The results of individual pairwise comparisons of physiological parameters before and after exposure to the “Serpisten” substance were processed. The significance of the differences was assessed using Student's t-test.

Example 1. In mice, the actoprotective effect of the substance "Serpisten" on increasing the dynamic and static performance of animals is studied. In immature mice at a dose of 5 mg / kg, swimming endurance (dynamic performance) increased by 80%. In adult white mice at a dose of 10 mg / kg, endurance increased on average by 63%, and in CBA mice by 107%. Static performance (load retention) under the influence of the substance "Serpisten" increased on average by 42%. Increased endurance to extreme physical exertion persists in animals and 7 days after cessation of drug administration.

Example 2. In mice study the central nervous system-tonic effect of the substance "Serpisten". Despite the available dose, interspecific, and individual differences in sensitivity to the action of "sickle," we observed a high central nervous system tonic effect of the drug in both sexually and immature mice. An acceleration of the orientational-research reaction and memory stimulation were found in animals that received the Serpisten substance, which was expressed in enhancing the search, remembering the route and accelerating the way out of the maze (“mental performance”). An increase in central nervous system tone developed simultaneously with an increase in physical performance in animals after administration of the drug (coefficient corr. Equal to 0.58). Under the influence of Serpisten at a dose of 50 mg / kg, the maze time in adult animals was reduced by 40%, and in immature white mice, with a total dose of 5 mg / kg, the maze was doubled compared to the control (p <0.05) .

Serpisten is currently undergoing clinical trials. Pharmacopoeia articles on raw materials, substance and dosage form are issued.

Claims (1)

The use of a mixture of phytoecdysteroids 20-hydroxyecdysone and 25S-inocosterone isolated from the aerial parts of plants of the genus Serratula of the Asteraceae family, preferably leaves collected during the vegetation phase, mass budding or the beginning of flowering, as a tonic and actoprotective agent.