Method for producing agent showing anticancer activity, and agent produced by this method (versions)

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to chemical-pharmaceutical industry. The developed versions of the method for producing the agent showing anticancer activity enables basidiomycete cultivation shorter from 2-3 weeks to 7 days and higher mycelium yield due to developing a composition of a nutrient medium, cultivation environment, sowing mycelium preparation, as well as higher efficacy of the agent. The method for producing the agent provides preparing the sowing basidiomycetes mycelium, preparing a processing medium, sowing the processing nutrient medium containing carbon and nitrogen sources, mineral salts with the prepared sowing mycelium, culturing basidiomycete to produce a submerged culture and recovering the agent showing anticancer activity. The sowing basidiomycetes mycelium is prepared in two stages, first of which is executed on a dense nutrient medium containing a corn extract, wheat water and agar, while the second one - on a fluid medium containing glucose, soya flour, potassium dihydrophosphate, magnesium sulphate and arachidonic acid. The prepared processing medium contains vegetable oil and soya flour in amounts 10-27 g/l and 16-27 g/l of water respectively, as well as potassium dihydrophosphate and magnesium sulphate in amounts 2.0-3.5 g/l and 0.2-0.4 g/l respectively. The basidiomycetes used are Ganoderma lucidum (Curt.:Fr.) P.Karst or Hypsizygus ulmarius (Bull.) Redhead, or Trametes versicolor (L.:Fr.) Pilat. The agent showing anticancer activity is recovered from the submerged culture prepared by cultivation or of the basidiomycete mycelium. The efficacy of the anticancer agent expressed by tumour growth inhibition in experiments in vivo makes 43-83%.

EFFECT: creation of the agent showing anticancer activity refers to the method for producing said agent by basidiomycetes cultivation in the submerged culture.

24 cl, 1 tbl, 22 ex

 

The invention relates to medicine, namely to obtain funds with antitumor activity by cultivation of basidiomycetes in submerged culture.

It is known that metabolites of many species of basidiomycetes have medicinal properties, in particular anti-tumor activity, including direct inhibitory effect on the proliferation and growth of tumor cells, enhancing antitumor immunity, anticarcinogenic effect and others, antioxidant, antiviral, hepatoprotective, immunomodulatory effects. In recent years in Russia, USA, China, Japan, Korea are working on obtaining medicines on the basis of basidiomycetes. The development of new drugs is constrained by the complex expensive technology of cultivation of basidiomycetes and excretion of these substances with stable biological properties, in particular anticancer.

Education basidiomycetes valuable for pharmacology of substances depends on the used species of basidiomycetes and their cultivation (preparation of inoculum, the cultivation conditions), and the process of excretion of metabolites.

For the treatment of cancer using chemical substances such as drugs having a high toxicity that og is unicipal their application.

Toxicity of anticancer agents derived from edible or non-edible (non-poisonous) basidiomycetes according to literature data, or no, or very low (Chang &But, 1986; Su et al., 1987).

Currently, much attention is paid to the development of the technology of cultivation of basidiomycetes with the aim of obtaining the maximum number of mycelium under cultivation in the shortest possible time and methods selection of anticancer agents.

A method of obtaining funds with antitumor activity, from the mycelium of the fungus Ganoderma lucidum. The mycelium is produced by cultivation of Ganoderma lucidum in submerged culture on a nutrient medium containing 50 g glucose, 20 g peptone, 0.87 g of potassium dihydrophosphate, 0.5 g of magnesium sulfate, 10 mg of iron chloride (II), 7 mg of manganese chloride, 10 mg of zinc sulfate, 4 mg of zinc chloride per 1 liter of water, pH of 5.5.

Pre-prepared seed. For that particular mushroom is cultivated at 180 rpm and 25°C for 10 days to obtain pellets obtained culture subcultured in fresh medium in an amount of 5% and re-cultured for 10 days. Obtained in two stages seeds sown fresh medium and cultured under the same conditions for 7 days. The mycelium is separated from the culture fluid and from it produce proteoglycan G009 - cf is a rotary, with antitumor activity, clean it and determine the antitumor activity against sarcoma 180, vaccinated male mice (EN 2082755 C1, 19.08.1997).

The disadvantage of this method is the long process of preparation of inoculum (up to 20 days), prolonged culturing of the fungus (7 days) and the complex process of allocating funds with antitumor activity.

A method of obtaining funds with antitumor activity from the mycelium of basidiomycetes selected from the group Ganoderma lucidum, Hypsizygus ulmarium, Kuhneromyces mutabilis, Omphalotus olearius, Panus conchatus, Piptoporus betulinus, Pleurotus eryngii, Trametes zonata.

The basidiomycetes first grown on agar medium at 27°C and then subcultured in liquid medium and cultivated in submerged culture at 27°C, 180 rpm for 2-3 weeks. Medium containing 2% glucose, 0.1% peptone, 0.1% of yeast extract, 0.1% of potassium dihydrophosphate, 0.1% magnesium sulfate and trace elements (iron sulfate (II), manganese sulfate, zinc sulfate, copper sulfate). From submerged culture produce mycelium, dried and dry mycelium is extracted with a means of having antitumor activity. As extractant use methanol, ethanol, acetonitrile, ethyl acetate, chloroform, dichloromethane, or a mixture thereof (US 20060057157, 16.03.2006).

The disadvantage of this method is perceived by the Oia funds with antitumor activity, is a long-term cultivation of basidiomycetes (2-3 weeks) and low efficiency of the funds received in the in vitro system. Antitumor activity of extracts from the mycelium of basidiomycetes conducted on cells myelogenous leukemia human K 562 line cells accounted for 53.8-69,0%. In the system in vivo received means tested were not.

The challenge aimed invention is the reduction in the duration of the process of cultivation of the basidiomycetes, increase the yield of mycelium and more effective anti-cancer agents.

The task in the part of the first variant of the invention is solved due to the fact that the method of obtaining funds with antitumor activity, comprising preparing a seed mycelium of the basidiomycete, preparing the production environment, sowing production nutrient medium containing sources of carbon, nitrogen, mineral salts, prepared seed mycelium cultivation of basidiomycete then get immersed in the culture, according to the invention in the preparation of production nutrient medium as a carbon source, use vegetable oil as a source of nitrogen in soybean flour in amounts, respectively 10-27 and 16 to 27 g/l of water, and in which the quality of mineral salts used potassium dihydrophosphate and magnesium sulfate in quantities respectively equal to 2.0 to 3.0 and 0.2-0.3 g/l of water.

As vegetable oils can be used sunflower, olive, soybean, linseed, peanut butter, or a mixture thereof.

The introduction of the protection of vegetable oil in the claimed amount contributes not only to shorten the duration of the cultivation process and increase the yield of mycelium, but also reduces the process of foam formation during cultivation.

Soy flour can be used from soybean varieties as genetically modified and unmodified.

Preparation of inoculum of basidiomycete can be carried out in two stages, the first of which is on a sterile nutrient medium containing corn steep 10-15 ml, agar-agar - 10-20 g and a decoction of wheat grains up to 1 liter, and the second sterile liquid nutrient medium containing (g/l): glucose and soy flour in amounts, respectively 17-23 and 8-15, potassium dihydrophosphate of 2.0 to 3.0, the magnesium sulfate of 0.2-0.4, water to 1 liter, at a temperature of 24-30°C for 2-4 days with aeration when this liquid nutrient medium before sterilization was incubated for 8-12 hours at room temperature.

From basidiomycetes it is advisable to use Ganoderma lucidum (Curt.:Fr.) P.Karst or Hypsizygus ulmarius (Bull.) Redhead, or Trametes versicolor (L.: Fr.) Pilat.

After culturing the obtained submerged culture can is about to dry and use as antitumor agents.

From the obtained after cultivation submerged culture, it is recommended to separate the mycelium, dried at a temperature of 30-90°C., crushed to a powder and the resulting powder to use as a tool with antitumor activity.

Selection means having antitumor activity, can be carried out by obtaining an aqueous extract of dried powdered mycelium taken in the amount of 18-100 g/l of boiling water for 2-3 hours at 1.0-1.5 ATM.

In the resulting aqueous extract can be added ethanol in the volume ratio 1:2-4, separating the precipitate and dry it, and use it as a tool with antitumor activity.

Obtained after cultivation of basidiomycete submerged culture should be autoclaved for 2-3 hours at 1.0-1.5 ATM, to separate the liquid phase and to use it as a tool with antitumor activity.

The liquid phase obtained after autoclaving submerged culture, it is advisable to dry and use as antitumor agents.

Antitumor agent obtained in the described manner, is an independent object of the invention.

The task in the part of the second variant of the invention is solved by h is about the method of obtaining funds with antitumor activity, comprising preparing a seed mycelium of each of the basidiomycetes, the preparation of sterile production medium, seeding production nutrient medium containing sources of carbon, nitrogen, mineral salts, prepared seed mycelium cultivation of basidiomycetes and allocation of funds, with antitumor activity, according to the invention carry out the cultivation of basidiomycetes species such as Ganoderma lucidum (Curt.:Fr.) P.Karst, Hypsizygus ulmarius (Bull.) Redhead, Trametes versicolor (L.: Fr.) Pilat, with each of these basidiomycetes cultured separately, prepare a production medium containing (g/l)as carbon source is vegetable oil as a source of nitrogen in soybean flour in equal amounts, respectively 10-27 and 16-27, as mineral salts of potassium dihydrophosphate and magnesium sulfate in amounts, respectively, of 2.0 to 3.0 and 0.2-0.3, and water to 1 liter.

As vegetable oils can be used sunflower, olive, soybean, linseed, peanut butter, or a mixture thereof.

The introduction of the protection of vegetable oil in the claimed amount contributes not only to shorten the duration of the cultivation process and increase the yield of mycelium, but also reduces the process of foam formation during cultivation./p>

Soy flour can be used from soybean varieties as genetically modified and unmodified.

Preparation of seed mycelium of each of these species of basidiomycetes can be carried out in two stages, the first of which is on a sterile nutrient medium, a liter which contains (in 1 liter of medium): corn steep 10-15 ml, agar - 10-20 g and a decoction of wheat grains up to 1 liter, and the second sterile liquid nutrient medium containing (g/l): glucose and soy flour in amounts, respectively 17-23 and 8-15, potassium dihydrophosphate - 2,0-3,0, magnesium sulfate - 0,2-0,4, water to 1 liter at a temperature of 24-30°C for 2-4 days with aeration. Before sterilization production nutrient medium suitable to stand at room temperature for 8-12 hours.

When the cultivation of basidiomycete Ganoderma lucidum in production nutrient medium should be introduced (g/l): vegetable oil and soy flour in amounts, respectively, equal to 10-17 and 16-20, additional carbon source in the form of glucose - 8-9, potassium dihydrophosphate - 2,0-2,5, magnesium sulfate - 0,25-0,30, water to 1 liter, and the cultivation of it is advisable to carry out at temperatures of 28-35°C for 3-5 days.

When the cultivation of basidiomycete Hypsizygus ulmarius in production nutrient medium it is recommended to enter (g/l): vegetable oil and soy is th flour in quantities respectively 20-27 and 20-27, potassium dihydrophosphate - 2,0-2,5, magnesium sulfate - 0,25-0,3, water to 1 liter, and the cultivation of it is advisable to exercise at a temperature of 25-32°C for 2-4 days.

Under cultivation of basidiomycete Trametes versicolor in production nutrient medium should be introduced (g/l): vegetable oil and soy flour in amounts, respectively 18-22 and 20-27, potassium dihydrophosphate - 2,5-3,0, magnesium sulfate - 0,20-0,25, water to 1 liter, and the cultivation is recommended to be performed at a temperature of 25-32°C for 3-4 days.

It is advisable mycelium of each basidiomycete separately dried at a temperature of 30-90°C., chop, take in the amount of 18-100 g/l and extracted with boiling water for 2-3 hours at 1.0 to 1.5 ATM, the resulting aqueous extracts were separated from the mycelium, mix in the ratio 0,5-1,0:0,5-1,0:0,5-1,0 and to use as a tool with antitumor activity.

It is recommended that the mycelium of each basidiomycete separately to dry, grind, take in quantities 18-100 g/l and extracted with boiling water for 2-3 hours at 1.0 to 1.5 ATM, to separate the aqueous extracts from the mycelium, and then each of the resulting aqueous extracts to introduce ethanol in the volume ratio 1:2-4, to separate each of the precipitate formed from the liquid phase, dried, mixed precipitation received in soo is wearing 0,5-1,0:0,5-1,0:0,5-1,0 and use the resulting mixture as a means of with antitumor activity.

The mycelium of each basidiomycete, separated from submerged culture, should be dried at a temperature of 30-90°C., crushed to a powder, mix in the ratio 0,5-1,0:0,5-1,0:0,5-1,0 and to use as a tool with antitumor activity.

A mixture of dry powdered mycelium of basidiomycetes, taken in an amount 18-100 g/l of water can be extracted with boiling water for 2-3 hours at 1.0 to 1.5 ATM, thus obtaining an aqueous extract, which can be used as a tool with antitumor activity.

It is recommended that in the resulting aqueous extract to introduce ethanol in the volume ratio 1:2-4, separating the precipitate, dry it, and use it as a tool with antitumor activity.

Appropriate after separation of the mycelium of each basidiomycete from submerged culture dried at 30 to 90°C, crushed to a powder, proektirovanii boiling water at a ratio of 18-100 g/l for 2-3 hours at 1.0-1.5 ATM, mix two of the resulting aqueous extracts of any basidiomycetes: Trametes versicolor and Hypsizygus ulmarius(1) or Trametes versicolor and Ganoderma lucidum (2), or Hypsizygus ulmarius and Ganoderma lucidum (3) in a ratio of 0.5 to 1.0:0.5 to 1.0, in the third extract of Ganoderma lucidum or Hypsizygus ulmarius, or Trametes versicolor add etano the in the volume ratio 1:2-4, the precipitate dried, add it to the mixture of aqueous extracts (1), (2) or (3), respectively, in an amount of 0.01-0.03% and to use as a tool with antitumor activity.

It is recommended that after the separation of the mycelium of each basidiomycete dried at 30 to 90°C, crushed to a powder, proektirovanii boiling water at a ratio of 18-100 g/l for 2-3 hours at 1.0-1.5 ATM, two of the obtained aqueous extracts of any basidiomycetes: Trametes versicolor and Hypsizygus ulmarius(1) or Trametes versicolor and Ganoderma lucidum (2), or Hypsizygus ulmarius and Ganoderma lucidum (3) add ethanol in each extract in the ratio of 1:2-4, the obtained precipitates were dried, mixed in a ratio of 0.5 to 1.0:0.5 to 1.0, to dissolve the mixture in the third extract of Ganoderma lucidum or Hypsizygus ulmarius, or Trametes versicolor in an amount of 0.01-0.03% and to use as a tool with antitumor activity.

Antitumor agent, received the second option described is an independent object of the invention.

As follows from the above, an antitumor agent can be used in a dry powder form and liquid. To the resulting antitumor agent may be added various pharmaceutical additives such as vitamins, minerals, antioxidants, compatible antitumor agents.

P is tivaouane the tool may be made in the form of tablets, capsules, drink and can be used per os, microclysters.

The antineoplastic dosage depends on the disease, patient's age, weight, composition tools. In terms of the content of water-soluble polysaccharides preferred dosage for laboratory animals can be 1-3 mg of polysaccharides per kg of weight for a person to 0.08-0.25 mg/kg

The technical result is to reduce the time consuming process of cultivation of basidiomycete from 2-3 weeks up to one and increasing the yield of mycelium through the development of the production environment, cultivation conditions, and also due to selection of medium composition for obtaining seed. The inclusion of the production environment of vegetable oil prevented the foaming medium during cultivation of basidiomycete.

In addition, the developed technology selection process antineoplastic agents allowed to obtain an effective remedy, which inhibits tumor growth in in vivo studies to 83%.

The invention is illustrated by the following examples, which do not cover the whole scope of the claims, but do not restrict it.

Example 1. Preparation of seed mycelium of the basidiomycete Ganoderma lucidum.

Agar sterile medium were prepared on the basis of the decoction of wheat, which was added kukuruz the second extract, 10 ml/l and agar 20 g/L. The medium was poured in test tubes and sterilized and seeded with a piece of mycelium of the basidiomycete Ganoderma lucidum strain G1-3. The mycelium was grown for 7 days.

Was prepared in a liquid nutrient medium containing (g/l): glucose - 22, soy flour - 10, potassium dihydrophosphate - 2.0 g/l, magnesium sulfate 0.3 g/l water to 1 liter. The prepared environment is kept for 10 hours at room temperature, and then sterilized.

Prepared liquid medium were seeded pre-grown on agar medium mycelia of G. lucidum. Cultivation was carried out at 30°C for 3 days, resulting in the growing mycelium.

Example 2. Cultivation of basidiomycete Ganoderma lucidum.

Was prepared sterile production medium containing (g/l): sunflower oil - 12, soy flour - 16, glucose - 9, potassium dihydrophosphate - 2,2, magnesium sulfate - 0.3, and water to 1 liter. Wednesday was sowing seed mycelium of Ganoderma lucidum strain G1-3, prepared by the method described in example 1, were cultured in 750 ml flasks at a temperature of 32°C on a rotary shaker at 180 rpm for 4 days and got submerged culture. The yield of air-dry biomass - 22 g/l

Example 3. Preparation of seed mycelium of the basidiomycete Hypsizygus ulmarius.

Agar sterile medium were prepared on the basis of the decoction of wheat, which was added corn extract 12 ml/l and the Aga is 10 g/L. The medium was poured in test tubes and sterilized and seeded with a piece of mycelium of the basidiomycete Hypsizygus ulmarius strain Hu-2. The mycelium was grown for 7 days.

Was prepared in a liquid nutrient medium containing (g/l): glucose - 17, soy flour, 8 potassium dihydrophosphate - 2,5, magnesium sulfate and 0.2, water to 1 liter. The prepared environment is kept for 8 hours at room temperature, and then sterilized.

Prepared liquid medium were seeded pre-grown on agar medium mycelium H.ulmarius. The culture of the fungus were grown at 24°C for 2 days, resulting in the growing mycelium.

Example 4. Cultivation of basidiomycete Hypsizygus ulmarius.

Was prepared sterile production medium containing (g/l): soybean oil - 26, soy flour - 26, potassium dihydrophosphate - 2,5, magnesium sulfate - 0,25, water to 1 liter. Wednesday was sowing seed mycelium Hypsizygus ulmarius Hu-2, prepared by the method described in example 3 was cultured in 750 ml flasks at a temperature of 28°C on rotary shaker at 230 rpm for 3 days. The yield of air-dry biomass was 39 g/L.

Example 5. Preparation of seed mycelium of the basidiomycete Trametes versicolor.

Agar sterile medium were prepared on the basis of the decoction of wheat, which was added corn extract 15 ml/l and agar 15 g/L. the Medium was poured in test tubes and sterilized and C is sevali a piece of mycelium of the basidiomycete Trametes versicolor strain Tv-12. The mycelium was grown for 7 days.

Was prepared in a liquid nutrient medium containing (g/l): glucose - 20, soya flour - 15, potassium dihydrophosphate - 3.0, magnesium sulfate - 0,4, water to 1 liter. The prepared environment is kept for 12 hours at room temperature, and then sterilized.

Prepared liquid medium were seeded pre-grown on a dense mycelium T.versicolor and cultivated at 26°C for 4 days, resulting in the growing mycelium.

Example 6. The cultivation of basidiomycetes Trametes versicolor.

Was prepared sterile production medium containing (g/l): olive oil - 19, soybean flour - 21, potassium dihydrophosphate - 2,5, magnesium sulfate - 0,25, water to 1 liter. Wednesday was sowing seeds Trametes versicolor Tv-12, prepared by the method described in example 5 were cultured in 750 ml flasks at a temperature of 26°C on a rotary shaker at 200 rpm for 3 days. The yield of air-dry biomass of 27 g/l

Example 7. Cultivation of basidiomycete Hypsizygus ulmarius in the bioreactor.

Cultivation was performed in a bioreactor with a volume of 1 m3the volume of the nutrient medium 300 liters of the Production medium contained (g/l): a mixture of vegetable oils (sunflower oil and soybean oil in the ratio 4:1) - 22, soy flour - 26, potassium dihydrophosphate - 2,0, magnesium sulfate and 0.2, the water - up to 1 is itra. Sterile medium were seeded seeds Hypsizygus ulmarius Hu-2, prepared according to the method described in example 2. The strain was grown in a bioreactor for 3 days at a temperature of 28°C, rotating paddle stirrer at 200 rpm, the air flow of 1.5 volume 1 volume of medium per minute. The yield of air-dry biomass was 35 g/l

Example 8. Receiving means having antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, 7, filtered, separated mycelium. The mycelium of each of the obtained culture was dried at 40°C, crushed to a powder and used as antitumor agents. The results presented in the table.

Example 9. Receiving means having antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, 7, filtered, separated mycelium. The mycelium of each of the obtained culture was dried at a temperature of 30°C, crushed to powder. The dry powder of each basidiomycete filled with distilled water at the rate of 30 g/l, autoclaved at 1.2 ATM for 2 hours, filtered and received water extracts each basidiomycete, which was used as antitumor agents. The results presented in the table.

Example 10. Receiving environments is TBA, with antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, was filtered, separated mycelium. The mycelium of each of the obtained culture was dried at 50°C, crushed to powder. The dry powder of each basidiomycete filled with distilled water, and the powder was taken in a quantity of 40 g/l, were extracted with water for 3 hours at 1.5 ATM, filtered, received water extracts. In each of the obtained water extract of mycelium of Ganoderma lucidum, Trametes versicolor, Hypsizygus ulmarius was administered ethanol in the ratio of 4 volumes of ethanol to 1 volume of the aqueous extract. The formed precipitation, representing the polysaccharide fraction was separated by centrifugation and liofilizirovanny, and thus, anti-cancer drugs. Antitumor properties of the funds studied separately. The results presented in the table.

Example 11. Receiving means having antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, autoclaved at 1.5 ATM for 3 hours, each separated liquid phase by filtration and received an antitumor agent in a liquid extract. A portion of each of the liquid extract was dried by spray drying, receiving antineoplastic agents in powder form. Efficiency recip is the R in liquid and dry form of anticancer agents presented in the table.

Example 12. Receiving means having antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, was filtered, separated mycelium. The mycelium of Ganoderma lucidum, Trametes versicolor, Hypsizygus ulmarius was dried at a temperature of 90°C, crushed to a powder, mixed in a ratio of 0.50:to 0.75:1.00 and used as antitumor agents. The results presented in the table.

Example 13. Receiving means having antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, was filtered, separated mycelium. The mycelium of Ganoderma lucidum, Trametes versicolor, Hypsizygus ulmarius was dried at a temperature of 30°C, crushed to a powder, mixed in a ratio of 1:1:1. The mixture of dry mycelium was flooded with distilled water, and the mycelium was taken in a quantity of 50 g/l, was extracted with boiling water at 1.2 ATM for 2 hours, filtered, received aqueous extract, which was used as antitumor agents. The results presented in the table.

Example 14. Receiving means having antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, was filtered, separated mycelium. The mycelium of Ganoderma lucidum, Trametes versicolor, Hypsizygus ulmarius was dried at 40°C, crushed to Poroskov asnago state, mixed in the ratio of 0.75:0,75:1,00. The mixture of dry mycelium was flooded with distilled water, and the mycelium was taken in the amount of 40 g/l, was extracted with boiling water at 1.3 ATM for 2 hours, filtered, received aqueous extract. In the resulting aqueous extract was added to ethanol at a rate of 1:2. The precipitate polysaccharide fraction was separated by centrifugation, liofilizirovanny and used as antitumor agents. The results presented in the table.

Example 15. Receiving means having antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, was filtered, separated mycelium. The mycelium of each of the obtained culture was dried at 40°C, crushed to powder. The dry powder of each basidiomycete filled with distilled water at the rate of 100 g/l, was extracted with boiling water at 1.0 ATM for 3 hours, filtered, received water extracts. The resulting aqueous extracts of the mycelium of Ganoderma lucidum, Trametes versicolor, Hypsizygus ulmarius was mixed in a ratio of 1:1:1 and used as antitumor agents. The results presented in the table.

Example 16. Receiving means having antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, shown is ovali, separated mycelium. The mycelium of each of the obtained culture was dried at a temperature of 30°C, crushed to powder. The dry powder of each basidiomycete filled with distilled water at the rate of 100 g/l, was extracted with boiling water at 1.0 ATM for 3 hours, filtered, received water extracts. In each of the obtained aqueous extracts of the mycelium was added ethanol in the ratio 1:4. The obtained precipitation of the polysaccharide fraction of Ganoderma lucidum, Trametes versicolor, Hypsizygus ulmarius was separated by centrifugation, liofilizirovanny and mixed in the ratio of 1.0:0.5 to 1.0 second and used as antitumor agents. The results presented in the table.

Example 17. Receiving means having antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, autoclaved at 1.5 ATM for 3 hours, filtered and the separated liquid phase. Thus obtained from submerged culture of Ganoderma lucidum, Trametes versicolor, Hypsizygus ulmarius liquid phase was mixed in a ratio of 1:1:1 and received an antitumor agent in liquid form. Portion of the liquid extract was dried in the spray dryer, receiving antitumor agent in powder form. The effectiveness of anti-cancer agents in liquid and dry form presented in the table.

Example 18. Receiving means having protivoopujolevy the howl activity.

Each submerged culture obtained in examples 2, 4, 6, was dried on the freeze dryer at core temperature change, including multiple temperature drops to minus 30°C. was Obtained antitumor agents in the form of a dry powder. Antitumor properties of the funds studied separately. The results presented in the table.

Example 19. Receiving means having antitumor activity.

Dry powder of freeze-dried submerged cultures of basidiomycetes, obtained in example 18, were combined in the ratio 1:1:1 and studied the antitumor properties of composite tools. The results presented in the table.

Example 20. Receiving means having antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, was filtered, separated mycelium. The mycelium of each of the obtained culture was dried at 45°C, crushed to powder. The dry powder of each basidiomycete filled with distilled water at the rate of 45 g/l, was extracted with boiling water at 1.5 ATM for 2 hours, filtered, received water extracts. Water extracts Trametes versicolor and Hypsizygus ulmarius was mixed in a ratio of 0.75:1.00 each. In water extract of mycelium of Ganoderma lucidum was added ethanol in the ratio 1:4. Poluchennykh Ganoderma lucidum, representing the polysaccharide fraction was separated by centrifugation, liofilizirovanny and was added in the amount of 0.02% in a mixture of extracts of Trametes versicolor and Hypsizygus ulmarius, receiving antitumor agent. The effectiveness of anti-cancer drugs are presented in the table.

Example 21. Receiving means having antitumor activity.

Each submerged culture obtained in examples 2, 4, 6, was filtered, separated mycelium. The mycelium of each of the obtained culture was dried at a temperature of 37°C, crushed to powder. The dry powder of each basidiomycete filled with distilled water at the rate of 18 g/l, was extracted with boiling water at 1.2 ATM for 3 hours, filtered, received water extracts of Ganoderma lucidum, Trametes versicolor, Hypsizygus ulmarius. In the extracts of Ganoderma lucidum and Trametes versicolor were added ethanol in the ratio 1:2. The obtained precipitation Ganoderma lucidum, Trametes versicolor, representing the polysaccharide fraction was separated by centrifugation, liofilizirovanny, mixed in the ratio of 1:0.5, it was added to the resulting mixture in an aqueous extract of Hypsizygus ulmarius in an amount of 0.01% was used as antitumor agents. The results presented in the table.

Example 22. Evaluation of antitumor activity of the funds received by the examples 8-21.

The funds received were experienced in the experience of the x in vivo in adult male mice-hybrids (C57B1/6×DBA/2)F 1(hereinafter B6D2F1with transplantable solid T-cell lymphoma R. Mice were obtained from nursery RAMS "Kryukovo". After receipt of mice kept in quarantine for 21 days. Lymphoma was supported in syngeneic conditions in ascitic form serial intraperitoneal passages in mice DBA/2. In the experience of the tumor was inoculated under the skin of the right-side 106cells at day "0". Treatment was started 48 hours after inoculation of the tumor. Received funds administered orally using vnutrisubstratnogo probe daily 1 time per day for 10 days. Dry powdered biomass of fungi and freeze-dried submerged cultures of the fungi were tested at a dose of 50 mg/kg/day, preparations were prepared by adding chopped mushroom material in starch paste so that the daily dose for each animal was kept in 0.3 ml of paste. The polysaccharide fraction was used in a dose of 2 mg/kg/day in the form of aqueous solutions, the daily dose of the solution was 0.3 ml/mouse. The dose of aqueous extracts of the mycelium of Ganoderma lucidum, Trametes versicolor and Hypsizygus ulmarius was 0.25 to 0.3 ml/kg/day, the solids content in water extracts ranged from 14 to 18 mg/ml Dried using spray drying aqueous extracts of the mycelium of fungi was tested at a dose of 4.8 mg/kg/day, sample dry extracts were dissolved in water, the volume of injected solution was 0.3 ml/m is nil. The number of mice in the control and experimental groups were 10 and 8 animals, respectively. During the experiments followed the General condition of the mice, the change of body weight, only the tumor, the dynamics of tumor growth. The tumor weight was calculated by the formula M (mg)=(a×b×C)/2, where M is the calculated tumor weight, a, b, C - 3 of the greatest perpendicular diameters of the tumor site in mm. Inhibition of tumor growth (TRO) was calculated by the formula: TRO (%)=(Mto-Mabout)/(Mto)×100, where Mtoand Mabout- the average weight of tumors in the control and experience respectively. The calculation was carried out according to figures obtained on the 14th day experience. The significance of differences of average values of tumor mass was determined by t-criterion Student. For reliable took differences at P≤0,05. The results are presented in the table. The table shows antitumor activity funds obtained from submerged cultures of the fungi.

Table
ExampleMedicationSRW, %
8The dry mycelium of Ganoderma lucidum (cultivation in flasks)51
Dry mycelium Tramees versicolor (cultivation in flasks) 65
Dry mycelium Hypsizygus ulmarius (cultivation in flasks)55
Dry mycelium Hypsizygus ulmarius (cultivation in the bioreactor)63
9Water extract of mycelium of Ganoderma lucidum57
Water extract of mycelium of Trametes versicolor56
Water extract of mycelium Hypsizygus ulmarius80
Water extract of mycelium of Hypsizygus ulmarius, grown in the bioreactor77
10A polysaccharide fraction of mycelium of Ganoderma lucidum71
A polysaccharide fraction of the mycelium of Trametes versicolor74
A polysaccharide fraction of mycelium Hypsizygus ulmarius72
11The liquid extract obtained from submerged culture of Ganoderma lucidum65
The liquid extract obtained from submerged culture of Trametes versicolor 56
The liquid extract obtained from submerged culture Hypsizygus ulmarius66
The dry extract obtained from submerged culture of Ganoderma lucidum62
The dry extract obtained from submerged culture of Trametes versicolor60
The dry extract obtained from submerged culture Hypsizygus ulmarius67
12The mixture of dry micelles Ganoderma lucidum, Trametes versicolor and Hypsizygus ulmarius60
13A water extract of a mixture of micelles Ganoderma lucidum, Trametes versicolor, Hypsizygus ulmarius73
14A polysaccharide fraction of a mixture of micelles Ganoderma lucidum, Trametes versicolor, Hypsizygus ulmarius, 2 mg/kg/day79
15The mixture of aqueous extracts of the mycelium of Ganoderma lucidum, Trametes versicolor and Hypsizygus ulmarius68
16A mixture of polysaccharide fractions of Ganoderma lucidum mycelium, Trametes versicolor and Hypsizygus ulmarius83
17The liquid mixture of extracts obtained from submerged cultures of Ganoderma lucidum, Trametes versicolor and Hypsizygus ulmarius68
The dried mixture of extracts obtained from submerged cultures of Ganoderma lucidum, Trametes versicolor and Hypsizygus ulmarius64
18Freeze-dried submerged culture of Ganoderma lucidum50
Freeze-dried submerged culture of Trametes versicolor67
Freeze-dried submerged culture Hypsizygus ulmarius43
19The mixture is freeze-dried submerged cultures of Ganoderma lucidum, Trametes versicolor, Hypsizygus ulmarius77
20The mixture of aqueous extracts of the mycelium of Trametes versicolor and Hypsizygus ulmarius and the polysaccharide fraction of the mycelium of Ganoderma lucidum79
21A mixture of polysaccharide fractions of Ganoderma lucidum mycelium and Trametes versicolor and water extract of mycelium Hypsizygus ulmarius75

From the presented data it follows that the reduced duration cooked what I seed mycelium from two weeks to 9-10 days, reduces the duration of the cultivation process to 2-4 days with a simultaneous increase in the yield of mycelium and effectiveness of the funds received with antitumor activity.

1. The method of obtaining funds with antitumor activity, comprising preparing a seed mycelium of the basidiomycete, preparing the production environment, sowing production nutrient medium containing sources of carbon, nitrogen, mineral salts, prepared seed mycelium cultivation of basidiomycete then get immersed in the culture, characterized in that in the preparation of production nutrient medium as a carbon source, use vegetable oil as a source of nitrogen in soybean flour in amounts, respectively 10-27 g/l and 16 to 27 g/l of water and mineral salts using potassium dihydrophosphate and magnesium sulfate in amounts respectively 2.0-3.0 g/l and 0.2-0.3 g/l of water.

2. The method according to claim 1, characterized in that the preparation of seed mycelium of the basidiomycete carried out in two stages, the first of which is on a sterile nutrient medium containing (in 1 liter of medium): corn steep 10-15 ml, agar - 10-20 g and a decoction of wheat grains up to 1 l of medium, and the second sterile liquid nutrient medium containing (g/l): glucose and soy mu is the in quantities respectively 17-23 and 8-15, potassium dihydrophosphate - 2,0-3,0, magnesium sulfate - 0,2-0,4, water up to 1 liter at a temperature of 24-30°C for 2-4 days with aeration, with the liquid nutrient medium before sterilization was incubated for 8-12 h at room temperature.

3. The method according to claim 1, characterized in that from basidiomycetes use of Ganoderma lucidum (Curt.:Fr.) P.Karst or Hypsizygus ulmarius (Bull.) Redhead, or Trametes versicolor (L.:Fr.) Pilat.

4. The method according to claim 1, characterized in that after culturing the obtained submerged culture is dried and used as antitumor agents.

5. The method according to claim 1, characterized in that after culturing obtained from submerged cultures separate the mycelium, dried at a temperature of 30-90°C., crushed to a powder, and the obtained dry powder is used as antitumor agents.

6. The method according to claim 5, characterized in that the dry powdered mycelium taken in the amount of 18-100 g/l, by extraction with boiling water for 2-3 h at 1.0 to 1.5 ATM receive water extract of mycelium used as a means with antitumor activity.

7. The method according to claim 6, characterized in that the resulting aqueous extract add ethanol in the volume ratio 1:2-4, separating the precipitate, dry it, and use as a tool with about ipooponyou activity.

8. The method according to claim 1, characterized in that obtained after cultivation of basidiomycete submerged culture autoclave for 2-3 h at 1.0 to 1.5 ATM, the liquid phase is separated and used as a means with antitumor activity.

9. The method according to claim 8, characterized in that the liquid phase obtained after autoclaving submerged culture, dried and used as a means with antitumor activity.

10. Antitumor agent obtained according to any one of claims 1 to 9.

11. The method of obtaining funds with antitumor activity, comprising preparing a seed mycelium of basidiomycetes, preparation of sterile production medium, seeding production nutrient medium containing sources of carbon, nitrogen, mineral salts, prepared seed mycelium cultivation of basidiomycetes, followed by the separation of the mycelium obtained from submerged cultures, characterized in that from basidiomycetes use basidiomycetes species of Ganoderma lucidum (Curt.:Fr.) P.Karst, Hypsizygus ulmarius (Bull.) Redhead, Trametes versicolor (L.:Fr.) Pilat, each of these basidiomycetes cultured separately, prepare a production medium containing (g/l)as carbon source is vegetable oil as a source of nitrogen in soybean flour in quantities sootvetstvenno 10-27 and 16-27, as mineral salts - potassium dihydrophosphate and magnesium sulfate in amounts respectively of 2.0 to 3.0 and 0.2-0.3, and water up to 1 L.

12. The method according to claim 11, characterized in that the preparation of seed mycelium of each of these basidiomycetes carried out in two stages, the first of which is on a sterile nutrient medium containing (in 1 liter of medium): corn steep 10-15 ml, agar - 10-20 g and a decoction of wheat grains up to 1 l of medium, and the second sterile liquid nutrient medium containing (g/l): glucose and soy flour in amounts, respectively 17-23 and 8-15, potassium dihydrophosphate - 2,0-3,0, magnesium sulfate - 0,2-0,4, water up to 1 liter at a temperature of 24-30°C for 2-4 days with aeration.

13. The method according to claim 11, characterized in that before sterilization production nutrient medium was incubated for 8-12 h at room temperature.

14. The method according to claim 11, characterized in that the cultivation of basidiomycete Ganoderma lucidum in production nutrient medium is injected (g/l): vegetable oil and soy flour in amounts, respectively, equal to 10-17 and 16-20, additional carbon source in the form of glucose - 8-9, potassium dihydrophosphate - 2,0-2,5, magnesium sulfate - 0,25-0,30, water to 1 l, and the cultivation is carried out at 28-35°C for 3-5 days.

15. The method according to claim 11, characterized in that the cultivation of basidiomycete Hypsizygus ulmarius in production is public medium is injected (g/l): vegetable oil and soy flour in quantities respectively 20-27 and 20-27, potassium dihydrophosphate - 2,0-2,5, magnesium sulfate - 0,25-0,30, water to 1 l, and the cultivation is carried out at a temperature of 25-32°C for 2-4 days.

16. The method according to claim 11, characterized in that the cultivation of basidiomycete Trametes versicolor in production nutrient medium is injected (g/l): vegetable oil and soy flour in amounts, respectively 18-22 and 20-27, potassium dihydrophosphate - 2,5-3,0, magnesium sulfate - 0,20-0,25, water to 1 l, and the cultivation is carried out at a temperature of 25-32°C for 3-4 days.

17. The method according to claim 11, characterized in that the mycelium of each basidiomycete separately dried at a temperature of 30-90°C., crushed, take in the amount of 18-100 g/l of water and extracted with boiling water for 2-3 h at 1.0 to 1.5 ATM, the resulting aqueous extracts are mixed in a ratio 0,5-1,0:0,5-1,0:0,5-1,0 and use as a tool with antitumor activity.

18. The method according to claim 11, characterized in that the mycelium of each basidiomycete separately dried, crushed, take in the amount of 18-100 g/l of water and extracted with boiling water for 2-3 h at 1.0 to 1.5 ATM, in each of the resulting aqueous extracts injected ethanol in the volume ratio 1:2-4, each of the precipitate formed is separated from the liquid phase, dried, mixed precipitation received in relation 0,5-1,0:0,5-1,0:0,5-1,0 and use in quality is TBE means, with antitumor activity.

19. The method according to claim 11, characterized in that the separated from submerged culture of the mycelium of each basidiomycete dried at a temperature of 30-90°C., crushed to a powder, mixed in a ratio 0,5-1,0:0,5-1,0:0,5-1,0 and the resulting mixture is used as a means with antitumor activity.

20. The method according to claim 19, characterized in that the mixture of dry powdered mycelium of basidiomycetes, taken in an amount 18-100 g/l of water, extracted with boiling water at a ratio of 2-3 h at 1.0 to 1.5 ATM, get a water extract, which is used as a means with antitumor activity.

21. The method according to claim 20, characterized in that the resulting aqueous extract is injected ethanol in the volume ratio 1:2-4, separating the precipitate, dried and used as a means with antitumor activity.

22. The method according to claim 11, characterized in that after separation of the mycelium of each basidiomycete from submerged culture, it is dried at a temperature of 30-90°C., crushed to a powder, take in the amount of 18-100 g/l), extracted with boiling water for 2-3 h at 1.0 to 1.5 ATM, the resulting aqueous extracts of any two basidiomycetes: Trametes versicolor and Hypsizygus ulmarius(1) or Trametes versicolor and Ganoderma lucidum (2), or Hypsizygus ulmarius and Ganoderma lucidum 3) are mixed in a ratio of 0.5 to 1.0:0.5 to 1.0, in the remaining third aqueous extract of Ganoderma lucidum or Hypsizygus ulmarius, or Trametes versicolor add ethanol in the volume ratio 1:2-4, the precipitate is dried and add it to the mixture of aqueous extracts (1), (2) or (3), respectively, in an amount of 0.01-0.03% and use as a tool with antitumor activity.

23. The method according to claim 11, characterized in that after separation of the mycelium of each basidiomycete from submerged culture it is dried at a temperature of 30-90°C., crushed to a powder, take in the amount of 18-100 g/l of water and extracted with boiling water for 2-3 h at 1.0 to 1.5 ATM, the resulting aqueous extracts of any two basidiomycetes: Trametes versicolor and Hypsizygus ulmarius (1) or Trametes versicolor and Ganoderma lucidum (2), or Hypsizygus ulmarius and Ganoderma lucidum (3) add ethanol in the volume ratio 1:2-4, the precipitate is dried, mixed in the ratio of 0.5 to 1.0:0.5 to 1.0, add it to the remaining third aqueous extract of Ganoderma lucidum or Hypsizygus ulmarius, or Trametes versicolor in an amount of 0.01-0.03% and use as a tool with antitumor activity.

24. Antitumor agent obtained according to any one of § § 11-23.



 

Same patents:

FIELD: medicine.

SUBSTANCE: invention relates to field of medicine and deals with anti-sense oligonucleotides for treatment and/or prevention of, at least, one of the following diseases: asthma, hypereosinophilia. Essence of the invention includes oligonucleotides aimed against sequences of nucleic acids, coding receptor, selected from group, which consists of receptor CCR3 and common subunit of receptors IL-3, IL-5 and GM-CSF with sequences SEQ ID NO:1 and SEQ ID NO:14.

EFFECT: reduction of toxicity in comparison with hormonal medications.

39 cl, 8 ex, 11 tbl, 21 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to composition for treatment of malignant neoplasm in patient, which contains epothilone - (1S, 3S, 7S, 10R, 11S, 12S, 16K)-7,11-dihydroxy-3-(2-methyl-benzotiasol-5-yl)-10-(prop-2-en-1-yl)-8,8,12,16-tetramethyl-4,17-dioxabicyclo[14.1.0]heptadecan-5,9-dion, hydroxypropyl-β-cyclodextrine or simple sulphobutyl ether of β-cyclodextrine and fillers, selected from mannitol and trometamol. Invention also relates to method of claimed composition obtaining, which includes dissolving epothilone in alcohol, dissolving hydroxypropyl-β-cyclodextrine or simple sulphobuthyl ether of β-cyclodextrine in water solution together with mannitol and trometamol, bringing pH of water solution to value 5-9 by means of hydrochloric acid and mixing obtained solutions.

EFFECT: invention also relates to method of malignant neoplasm treatment in patient, which includes intravenous infusion of claimed composition during period of approximately 30 minutes in dose from 10 mg/m2 to 35 mg/m2.

17 cl, 11 ex, 2 dwg

FIELD: medicine.

SUBSTANCE: invention concerns a method for detecting sensitivity of a biological sample containing human lung cancer cells, a combination of a epidermis growth factor receptor inhibitor and a chemotherapeutic agent, by analysing the biological sample for superexpression of phosphorylated ACT protein and phosphorylated MAPC protein.

EFFECT: extended range of methods for evaluating sensitivity to EGFR inhibitors therapy.

10 cl, 1 ex, 11 dwg, 3 tbl

FIELD: medicine.

SUBSTANCE: EGFL7 antibody is described. Offered is a pharmaceutical composition for vessel development modulation containing an effective amount of EGFL7 antibody. There are presented a polynucleotide coding said EGFL7 antibody, an expression vector containing said polynucleotide, a host cell able to express exogenous genetic material containing said vector. There is described a method of angiogenesis reduction or inhibition in a subject having a pathological condition associated with angiogenesis including the introduction to the subject of an effective amount of EGFL7 antibody or the pharmaceutical composition containing said antibody. There is offered a method for providing higher efficacy of an antiangiogenic agent in the subject having the pathological condition associated with angiogenesis including the introduction to the subject of EGFL7 antibody or the pharmaceutical composition containing said antibody.

EFFECT: invention extends the range of products available for vessel development modulation.

46 cl, 9 dwg, 2 tbl, 2 ex

Thienopyridines // 2415859

FIELD: chemistry.

SUBSTANCE: invention relates to pharmaceutically suitable salts which are given in claim 1. The invention also relates to medicinal agents based on the said compounds, which are HSP90 inhibitors.

EFFECT: novel compounds and medicinal agents based on said compounds are obtained, which can be used to treat diseases influenced by inhibition, regulation or modulation of HSP90.

3 cl, 1 tbl, 16 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula or pharmaceutically acceptable salt thereof; where Q is selected from a group consisting of R1 denotes H; each R2 independently denotes ZR; each JQ independently denotes ZQ, MQ, (LQ)-ZQ or (XQ)-MQ; each LQ and XQ independently denotes C1-6alkyl, where each carbon of the alkyl group is independently substituted in up to 2 cases -NR-, -NHR-, -NRC(O)O-; where each XQ is independently and optionally substituted with 0-2 JXQ; each ZR and ZQ independently denotes H; a C1-6alkyl group, a 5-7-member saturated or completely unsaturated monocyclic ring, having 0-2 nitrogen atoms; or a 9-member saturated bicyclic ring system having 1 nitrogen atom; where each is ZQ independently and optionally substituted with 0-3 JZQ; MQ denotes halogen, CN or N(R)2; each JLQ, JXQ and JZQ independently denotes V, M, (LV)-V, (LM)-M, halogen, C1-3alkoxy; each R independently denotes H, C1-6alkyl group; each LV and LM independently denotes C1-6alkyl, interrupted in up to 1 case by -C(O)-; where each V independently denotes H; C1-6alkyl group, a 5-6-member saturated or completely unsaturated monocyclic ring; where each V is independently or optionally substituted with 0-1 JV; each JV denotes NH2; each M independently denotes halogen, OH, O(C1-6alkyl), NH2, provided that when R1 and R2 - H, Q is not The invention also relates to a composition based compounds of formula I, a method of inhibiting activity of Aurora B protein kinase, FLT-3 protein kinase, a method of treating a proliferative disorder, particularly leukaemia or lymphoma and a method of producing compounds of formula I.

EFFECT: novel benzimidazole derivatives are obtained, which can be used as inhibitors of protein kinase Aurora B and FLT-3.

28 cl, 2 tbl, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to new compounds described by formula in which radical and symbol values are specified in the patent claim, and their pharmaceutically acceptable salts. These compounds inhibit tompomyosine-related kinases (Trk), and can find application in treating a malignant growth, such as breast cancer, rectal cancer and prostate cancer. Also, the invention relates to a method for producing these compounds, a based pharmaceutical composition and to methods of application thereof.

EFFECT: preparation of the pharmaceutical composition which can find application in treating a malignant growth.

18 cl, 134 ex

FIELD: chemistry.

SUBSTANCE: invention relates to an improved method of producing N-{5-[4-(4-methyl piperazinomethyl)benzoylamido]-2-methylphenyl)-4-(3-pyridyl)-2-pyrimidine amine of formula (I) (Imatibin) in form of a free base or acid addition salt. Said compounds have anti-tumour activity and can be used, for example, when treating leukaemia. The method involves reduction of N-(2-methyl-5-nitrophenyl)-4-(3-pyridyl)-2-pyrimidine amine of formula (IV) in the presence of a chemical reducing agent, reaction of the obtained N-(2-methyl-5-nitrophenyl)-4-(3-pyridyl)-2-pyrimidine amine of formula (II) with a dihydro-halide salt of 4-(4-methylpiperazinomethyl)benzoylhalide of formula (III) in the presence of an inert organic solvent which is not a base-acid acceptor to obtain a hydro-halide salt of imitinib of formula (I), where n equals 1, 2 or 3 and Hal denotes bromine, chlorine, fluorine or iodine in hydrated form, which, if needed, optionally undergoes further conversion to a free base or some other acid addition salt.

EFFECT: method simplifies the production and extraction process, the process takes place under mild conditions, the obtained hydrohalide salt of imatibin is virtually not soluble in organic solvents and can be easily extracted from the reaction mass.

39 cl, 3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to an improved method of producing N-{5-[4-(4-methyl piperazinomethyl)benzoylamido]-2-methylphenyl)-4-(3-pyridyl)-2-pyrimidine amine of formula (I) (Imatibin) in form of a free base or acid addition salt. Said compounds have anti-tumour activity and can be used, for example, when treating leukaemia. The method involves reduction of N-(2-methyl-5-nitrophenyl)-4-(3-pyridyl)-2-pyrimidine amine of formula (IV) in the presence of a chemical reducing agent, reaction of the obtained N-(2-methyl-5-nitrophenyl)-4-(3-pyridyl)-2-pyrimidine amine of formula (II) with a dihydro-halide salt of 4-(4-methylpiperazinomethyl)benzoylhalide of formula (III) in the presence of an inert organic solvent which is not a base-acid acceptor to obtain a hydro-halide salt of imitinib of formula (I), where n equals 1, 2 or 3 and Hal denotes bromine, chlorine, fluorine or iodine in hydrated form, which, if needed, optionally undergoes further conversion to a free base or some other acid addition salt.

EFFECT: method simplifies the production and extraction process, the process takes place under mild conditions, the obtained hydrohalide salt of imatibin is virtually not soluble in organic solvents and can be easily extracted from the reaction mass.

39 cl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new cyclopenta[b]benzofuranyl derivatives of formula wherein substitutes R1, R2, R3, R4, R5, R6 and R7 and n are specified in the patent clam. These compounds exhibit properties of NF-kB-activity and/or AP-1 inhibitor/modulator. Also, the inventive subject matter are methods for preparing intermediate compounds thereof, a pharmaceutical composition containing them, administration thereof for prevention and/or treatment of inflammatory and autoimmune diseases, neurodegenerative diseases and hyperproliferative diseases caused by NF-kB- and/or AP-1-activity, and a method for prevention and/or treatment of said diseases.

EFFECT: preparation of new cyclopenta[b]benzofuranyl derivatives.

21 cl, 3 tbl, 151 ex

FIELD: medicine.

SUBSTANCE: invention relates to cosmetology and represents antiperspirant composition, which contains carrier-substrate and water-soluble or dispersing in water tiomer and has nonaqueous dispersive phase with dispersed in it tiomer, where tiomer is product of polyethylenamine tioletion.

EFFECT: invention insures effective antiperspirant properties.

10 cl, 24 ex, 5 tbl

FIELD: medicine.

SUBSTANCE: invention relates to dentistry and represents composition for oral cavity care, which includes lactoferrine and lactoperoxydase, which differs in the fact that: it additionally contains licorice extract, lactoperoxydase activator and filler, and, composition components are in definite ratio in wt %.

EFFECT: invention insures high efficiency of inhibition of dental deposit formation.

18 cl, 8 ex, 12 tbl

FIELD: medicine.

SUBSTANCE: invention relates to dentistry and represents composition for oral cavity care, which includes lactoferrine and lactoperoxydase, which differs in the fact that: it additionally contains licorice extract, lactoperoxydase activator and filler, and, composition components are in definite ratio in wt %.

EFFECT: invention insures high efficiency of inhibition of dental deposit formation.

18 cl, 8 ex, 12 tbl

Product for peeling // 2416390

FIELD: medicine.

SUBSTANCE: invention relates to field of cosmetology and represents product for individual peeling, which contains a reservoir for individual application, into which placed are mixture-forming components, of oil phase, moistening additives, peeling, preservatives and water, which differ in the fact that: as peeling applied are microballs of silica, which have form close to spherical with size from 0.1 mcm to 20 mcm.

EFFECT: invention insures increase of safety and efficiency in application.

1 dwg

Product for peeling // 2416390

FIELD: medicine.

SUBSTANCE: invention relates to field of cosmetology and represents product for individual peeling, which contains a reservoir for individual application, into which placed are mixture-forming components, of oil phase, moistening additives, peeling, preservatives and water, which differ in the fact that: as peeling applied are microballs of silica, which have form close to spherical with size from 0.1 mcm to 20 mcm.

EFFECT: invention insures increase of safety and efficiency in application.

1 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to synthetic polymer chemistry. The nanocomposite contains a matrix in form of a cross-linked salt of hyaluronic acid which is modified with sulphur-containing compounds and nanoparticles of a noble metal as filler. A film of the cross-linked salt of hyaluronic acid which is modified with sulphur-containing compounds is obtained through chemical reaction of the salt of hyaluronic acid with a mixture of two sulphur-containing compounds and with a cross-linking agent, under conditions with pressure between 50 and 300 MPa and shear deformation in a mechanical reactor at temperature between 20 and 30°C. The reactor used to obtain the film is a Bridgman anvil.

EFFECT: invention enables to obtain a range of new bioactive nanocomposites with quantitative output and in the absence of a liquid medium, where the method does not require high energy, labour and water consumption and significantly increases efficiency of the composite; in particular, resistance to decomposition in the presence of hydroxyl radicals is 2-3 times higher compared to the control result.

16 cl, 7 ex

FIELD: medicine.

SUBSTANCE: invention relates to field of veterinary. Method consists in the following: applied is "EVL-Se-COMPOSITION" solution which is introduced into animals by group method with water one time per day in single dose 0.05 ml during 10 days, 5 days before weaning and 5 days after weaning.

EFFECT: application of claimed method makes it possible to realise prophylaxis of gastrointestinal diseases, increase average daily weight gain, does not cause side effects.

3 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to pharmacology and haematology. A hemostimulant represents hyaluronidase immobilised on low-molecular polyethylene glycol of molecular weight 400 to 4000 Da by directed accelerated electron flow 2.5 MeV with absorbed dose 2 to 10 kGy and dose setup rate 1.65 kGy/hour. For hemopoiesis stimulation, hyaluronidase immobilised as stated above is introduced either parenterally, or orally 1 time a day for 1-5 days in dose 10-1000 Unit/kg.

EFFECT: use of hyaluronidase immobilised with using an electron-beam synthesis technology enables higher safety and efficiency of hemopoiesis stimulation.

2 cl, 3 tbl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, particularly to a method for producing an agent for treating hysteromyoma. The method for producing the agent for treating hysteromyoma by vegetable oil extraction of crude drug containing Hungarian monkshood (root), Atragene sibirica L. (herb), bugbane (root), redhaw hawthorn (fruits), meadow clover (blossom), Paeonia anomala L. (root), Ramischia secunda (herb). The method of treating hysteromyoma.

EFFECT: method enables producing an effective agent for treating hysteromyoma.

2 cl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, particularly to a method for producing an agent for treating hysteromyoma. The method for producing the agent for treating hysteromyoma by vegetable oil extraction of crude drug containing Hungarian monkshood (root), Atragene sibirica L. (herb), bugbane (root), redhaw hawthorn (fruits), meadow clover (blossom), Paeonia anomala L. (root), Ramischia secunda (herb). The method of treating hysteromyoma.

EFFECT: method enables producing an effective agent for treating hysteromyoma.

2 cl, 3 ex

FIELD: medicine.

SUBSTANCE: invention relates to cosmetology and represents antiperspirant composition, which contains carrier-substrate and water-soluble or dispersing in water tiomer and has nonaqueous dispersive phase with dispersed in it tiomer, where tiomer is product of polyethylenamine tioletion.

EFFECT: invention insures effective antiperspirant properties.

10 cl, 24 ex, 5 tbl

Up!