Dry fucus extract, method for preparing it, and base anticoagulant ointment

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely an agent possessing anticoagulant activity. A method for preparing a dry fucus extract possessing anticoagulant action by the complex treatment of Fucus vesiculosus. The dry fucus extract possessing anticoagulant activity representing a polysaccharide complex containing fucoidan having the following composition: neutral monosaccharides - fucose, xylose, mannose, galacose, glucose, uronic acid; as well as polyphenols and sulphates in specific amount. The anticoagulant ointment containing the dry fucus extract.

EFFECT: agents described above possess pronounced anticoagulant action.

4 cl, 4 ex

 

The invention relates to medical and therapeutic practices, chemical-pharmaceutical, cosmetic industry, including the development of anticoagulants vegetable origin.

A method of obtaining the extract fukunaga by RF patent for the invention №2126688 from 27.02.1999, the Method consists of grinding fucus algae, extraction and filtration of the extract. Before extraction of algae treated with hot water, extraction is carried out in alkaline solution of soda ash in alkalinity of 0.7-0.8%, a temperature of 65±5°C for 4-5 h After filtration, evaporated extract and dried when the steam pressure is 0.19±0.01 MPa. The extraction of alkaline solutions fucus extract mainly contains alginates, with this method extract extract fucoidan does not occur. An extract of such a composition cannot be used as an anticoagulant drug.

It is known tool that has anticoagulant and immunotropic action for Russian patent for invention №2247574 from 26.12.2002, It is an extract obtained by extraction with water brown seaweed Fucus evanescens at a temperature of 20-60°C with subsequent precipitation with ethanol. The tool has a molecular mass of 20000-40000 daltons and contains a neutral sugars: fucose - 70-80%, galactose - 2-9%, xylose - 5-10% of the glucose - 2-7%, and-sulfate - 20-22%. Anticoagulant effect of the claimed fucoidan from Fucus evanescens is characterized by the following parameters when tested in vitro: activated partial thromboplastin time (APTT) from 80 to 900 C, thrombin time (TV) from 40 to 400, depending on the dose. On the anticoagulant action it is comparable to heparin, which also has a dose-dependent effect. Fucoidan receive the following way. Fresh or frozen (after depascale) algae treated with ethanol for degreasing and pigment removal. Then the seaweed is dried in the air, roughly pulverized and treated with water (1:20 wt/vol) at room temperature (5 hours), and then at 50-60°C under stirring for 5 hours. The extract was concentrated under reduced pressure to 1/4-1/5 of the initial volume and bring the pH of the suspension to a value of 3.0. Then separated from the precipitate by centrifugation, adjusted the pH of the supernatant to a neutral titration with sodium hydroxide, dialist against water. Then wikidot sodium salt of fucoidan two volumes of 96% ethanol. The precipitate three times washed with 50% aqueous ethanol and dried in 96% ethanol. Get the target product with the output of 7.5-10% (by weight of dry algae). Received fucoidan is a cream coloured powder, soluble in water, dimethyl sulfoxide, nearest the orim in alcohol, acetone, hexane, sulfuric ether. A study was conducted anticoagulant action of fucoidan: effect of fucoidan on the indices of coagulation tests in vivo and in vitro, which showed that the optimal concentration of fucoidan for the manifestation of maximum anticoagulant effect is in the range of 50-100 μg/ml Similar dose-dependent effect has, as is well known, and heparin. A feature of this method is the use as raw materials of the far Eastern algae F. evanescens containing 7-10% of fucoidan certain structure and composition, depending on the pharmacological properties of the polysaccharide. In addition, the application of the extraction of raw materials, inefficient methods of maceration (soaking) reduce process yields of target products, in particular of fucoidan. The deposition of alcohol is also accompanied by large losses of final products. Anticoagulant action of fucoidan from seaweed F. evanescens was studied by intravenous injection of the drug, its topical application was not considered.

The closest way to extract fucus is a method for complex processing of fucus vesiculosus by RF patent for the invention №2337571 from 11.08.2006, the Method is dry thallus algae are crushed, extracted with a mixture of methylene chloride with ethyl alcohol in which the rate was 94.2:5,86% to conventional Soxhlet extractions, concentrated in vacuo on a rotary evaporator, dried in vacuum on a rotary evaporator, dried in a vacuum drying Cabinet with receiving lipid-pigment complex - thick extract, then the meal is extracted by percolation method 85-90%ethanol with forced circulation of the extractant every hour infusion for 3-5 h, filtered, concentrated in vacuo on a rotary evaporator, crystallized, filtered on a suction filter, dried and purified by the method of column chromatography using alumina and receive mannitol cleared, then the remaining meal is extracted by the method of circulating the percolation 5-15%aqueous ethanol at pH 1-2 and time of infusion 6-9 hours at a temperature of 40-60°C, concentrated in vacuo on a rotary evaporator, dried in a vacuum drying Cabinet and get polysaccharide complex - dry extract of fucus, then the meal is extracted with a 1.5%solution of sodium carbonate is treated with sulfuric acid, concentrated on a rotary evaporator, and purified by dialysis through a cellophane membrane is dried in a vacuum drying Cabinet or by lyophilization and get the polysaccharide alginate sodium. The disadvantage of this method is the length of the extraction process, low pH (or high acidic solutions extractant), which can lead to hydrolysis of the final product - fucoidan, as well as strong the second heating of the extractant, requiring significant energy cost when extracting fucoidan.

The closest anticoagulant is heparin ointment. Pharmacological properties: Anticoagulant drug, has local anti-inflammatory and antiexudative action. Warns local thrombus formation, increases vascular permeability; accelerates resorption of hematomas; promotes the repair of damaged connective tissue and cartilage cells. Indications: thrombophlebitis, phlebitis superficial veins, periferic varicose ulcers, varicose ulcers, venous leg ulcers; status after phlebosclerosis, phlebectomy; furuncle, carbuncle, lymphadenitis; hematoma, contusions, the divergence of the old post-operative joints, to improve the healing of joints after trauma, burns, surgery; osteoarthritis of the knee and finger joints; chondropathy kneecap; aseptic infiltration, local swelling. Contraindications: hypersensitivity, hemorrhagic diathesis, different forms of purple, thrombocytopenia, hemophilia, tendency to bleeding, pregnancy. Side effects: allergic reactions (skin rash, itching, urticaria, angioedema). When applied to large surfaces possible hemorrhagic complications (due to resorptive action).

Dry EXT the act of fucus, as is known, is a polysaccharide complex, containing fucoidan.

A method of obtaining a dry extract of fucus by complex processing of fucus vesiculosus, as known, is that dry thallus algae are crushed, extracted with a mixture of methylene chloride with ethyl alcohol in the ratio was 94.2:5,86% to conventional Soxhlet extractions, concentrated in vacuo on a rotary evaporator, dried in vacuum on a rotary evaporator, dried in a vacuum drying Cabinet with receiving lipid-pigment complex - thick extract, then the meal is extracted by percolation method 85-90%ethanol with forced circulation of the extractant every hour infusion for 3-5 h, filtered, concentrated in vacuum on a rotary evaporator, crystallized, filtered on a suction filter, dried and purified by the method of column chromatography using alumina and receive mannitol cleared, then the remaining meal is extracted by the method of percolation 5-15%aqueous ethanol, concentrated and dried, yielding polysaccharide complex - dry extract of fucus, then the meal is extracted with a 1.5%solution of sodium carbonate is treated with sulfuric acid, concentrated on a rotary evaporator, and purified by dialysis through a cellophane membrane is dried in a vacuum drying Cabinet or by lyophilization and receive p is lished sodium alginate.

Ointment anticoagulant, as known, includes active and auxiliary substances, which use purified water, emulsifier and preservatives.

The objective of the invention is the improvement of complex processing of fucus algae in the Barents sea, including fucus vesiculosus, with the aim of expanding Arsenal of herbal medicines with anticoagulant action.

Technical results of the invention are: the acceleration of the extraction process and obtaining a dry extract of fucus with certain physico-chemical characteristics, enhancing its anticoagulant (heparinate) action, as well as receiving anticoagulant treatment-and-prophylactic means in the form of ointments on the basis of the obtained extract that reduce side effects while using that contribute to the expansion of anticoagulant means of plant origin.

The technical result according to the first claim is achieved by the fact, to obtain a dry extract of fucus possessing anticoagulant effect, algae meal is extracted with 4-6 hours at a temperature of 30-40°C and pH=3-4 percolation method with ultrasonic treatment, which is carried attached to the outside of percolator source of ultrasound, when Atomflota ultrasound produce for 15-20 minutes every hour infusion, then the extract is filtered, concentrated by ultrafiltration installation and dried in the spray dryer.

The technical result according to the second claim is achieved by the fact that the dry extract of fucus possessing anticoagulant effect obtained by the method according to claim 1, represents a polysaccharide complex, containing fucoidan following composition: neutral monosaccharides - fucose 41-44%, xylose 2-2,5%, mannose 3-3,5%, kalakota 4-4,5%, glucose 2%, uronic acids 3-3,5%; and polyphenols 18-20% and sulfates 21-25%.

The technical result for the third claim is achieved by the fact that the ointment anticoagulant includes dry extract of fucus according to claim 2, lanolin, olive oil, TWEEN 80, preservative, purified water, in the following ratio of components, wt.%:

dry extract of fucus - 0,15-30;

lanolin - 1-10;

olive oil - 10-30;

TWIN 80 - 1-5;

preservatives - 1-1,5;

purified water - the rest.

The ointment may contain the following ratio of components, wt.%:

dry extract of fucus - 0,15;

lanolin - 10;

olive oil - 10;

TWIN 80 - 1;

preservatives - 1;

purified water - the rest.

Fucus algae is a promising raw material for production of BAS. Stocks fucoidan on the coast of the Barents sea is estimated by experts as a significant (over 300 tons), and these species of algae have a wide advantage is defamiliarisation, grow in handy for harvesting locations in the intertidal zone. F. vesiculosus is representative of fucus on the coast of the Barents and White seas. Currently under extensive study anticoagulating properties of fucoidan (Raskin and others, 1991; Colliec et al.,1991; Nishino et al., 1991; Mauray et al., 1995; Giraux et al., 1998; Chevolot et al., 1999; Boissonvidal et al., 2000; Chevolot et al., 2001), which found that fucoidan affects blood rheology, like heparin. Heparin ointment is used as an anticoagulant drug, has local anti-inflammatory and antiexudative action. Warns local thrombus formation, increases vascular permeability; accelerates resorption of hematomas; promotes the repair of damaged connective tissue and cartilage cells. However, heparin is the drug of animal origin, and its application in the clinic can cause some side effects: reduction of blood coagulation, thrombocytopenia, the occurrence of gastrointestinal bleeding, allergic reactions and other Heparin poorly penetrates the intact skin and mucous membranes, due to this drug almost has a weak system resorptive effect. With long-term use on large surface possible hemorrhagic complications. Given the shortcomings of medicinal PR is parathas "heparin ointment", the actual search of new high-performance, low-toxic anticoagulants direct action of plant origin. The greatest recognition as such substances received connection polysaccharide nature, and in particular, algal polysaccharide fucoidan, and photoidentified dry extracts of algae.

Getting fucoidan most promising to be implemented in the integrated use of algal feedstock. Stage complex technology to obtain the thick extract and mannitol are both stages of cleaning in obtaining a dry extract of fucus.

Dry extract of fucus (SEF) in the nearest analogue is obtained from fucus algae meal of the Barents sea (F. vesiculosus, F. distichus, F. serratus, A. nodosum) within the complex technologies, which are as follows: dry thallus algae are crushed, extracted with a mixture of organic extractants, concentrated in vacuo on a rotary evaporator, dried in vacuum on a rotary evaporator, dried in a vacuum drying Cabinet with receiving lipid-pigment complex - thick extract, then successively extracted with ethanol, water-alcohol mixture and sodium hydroxide to obtain mannitol, dry extract and sodium alginate.

During the processing of fucus algae species of fucus vesiculosus for LIPI the but-pigment complex (dense fucus extract) extraction was carried out with a mixture of methylene chloride with ethyl alcohol in the ratio was 94.2:5.8% to conventional Soxhlet extractions. To obtain mannitol algal meal was extracted by percolation method 85-90% ethanol with forced circulation of the extractant every hour infusion for 3-5 hours. To obtain a dry extract of fucus remaining meal were extracted by the method of circulating the percolation 5-15%aqueous ethanol, pH=1-2. Time infusion 6-9 h, the temperature of 40-60°C. Then concentrated in vacuo on a rotary evaporator and dried in a vacuum drying Cabinet.

A method of obtaining a dry extract of fucus possessing anticoagulant effect, for the purposes of the present invention is as follows.

By the present method the meal algae were extracted 5-15%aqueous ethanol at pH=3-4 percolation method with ultrasonic treatment. The extraction time is 4-6 hours, the temperature of 30-40°C. the ultrasound Source was fixed on the housing of the extractor-percolator with its outer side. During infusion conducted the periodic (every hour) enable install on for 15-20 minutes Then the extract was filtered and concentrated by ultrafiltration installation. Dried in the spray dryer. The result is a dry extract of fucus is a powder from light brown to dark brown in color, contains fucoidan, polyphenols, easily soluble in water to form viscous solutions brown, physico-chemical features, the properties of which are shown in table 1.

However, with the intensification of the process of extraction of target components (polysaccharides, polyphenols and others), reducing the total time of extraction. Due to local heating during the ultrasonic treatment, the temperature of the extraction process can also be reduced. Reduced the total time of extraction, the reaction medium changed from acidic to slightly acidic, the optimal conditions of extraction time and temperature. Thus, the terms of the percolation fucus differ from similar method, temperature, pH, time. The result has been a change in the chemical composition of the active substances in the extract, and ultimately in the final product is a dry extract. Extraction with ultrasound accelerates the process of extraction of raw materials, providing a more complete extraction of active substances - polysaccharide fucoidan and polyphenols.

Emerging ultrasonic waves create alternating pressure, cavitation and sound the wind. The result is faster the swelling material and the dissolution of the cells, increases the speed of flow of the particles of raw materials, diffusion in the boundary layer is turbulent and vortical flows. Molecular diffusion inside the particles and diffusion in the boundary layer is almost replaced by convection, which then leads to the intensification of mass transfer. As a result of cavitation occurs the destruction of cell structures, which speeds up the process of transition of active substances in the extractant due to their leaching.

The use of ultrasound allows you to get the hood from several minutes to several hours depending on the efficiency of its use, which depends on process parameters: intensity and exposure scoring, selection of the extractant, the ratio of the raw material and the extractant and other optimal temperature when scoring no higher than 30-60°C, to avoid the formation of air bubbles, absorbing the ultrasonic waves. As extractant preferred spirtovanie mixture with a high concentration of ethanol, which inhibits the redox processes taking place in an ultrasonic field. For many commodities optimal intensity ultrasound (frequencies 2·104-2·108with-1) is in the range of 1.5 to 2.3·104W/m2. To obtain the claimed extract was used, the average intensity of ultrasound - 2·104W/m2. It is this combination of extraction and processing methods of extraction have led to a new product - polysacharide-polyphenolic dry extract of fucus, which is characterized by a specific chemical composition, technological properties, and, most importantly, the us is population anticoagulant actions in the composition of ointments.

Confirmation of the possibility of obtaining in this way the claimed technical result is a dry extract of fucus possessing anticoagulant effect is given in the following specific examples.

Example 1.

Dry powdered thallus of fucus vesiculosus were extracted by azeotrope of methylene chloride - ethyl alcohol to conventional Soxhlet extractions 6 o'clock Extract was concentrated, dried. Meal algae were extracted by the method of percolation with ethanol 85% at a temperature of 65°C, time of infusion, 5 o'clock Extract was filtered, concentrated in vacuo on a rotary evaporator. Was led at 0÷+4 C for 24 hours of Mannitol was filtered on a suction filter and dried, purified by the method of column chromatography using alumina. Meal algae were extracted by percolation method with ultrasonic treatment 5%aqueous ethanol, pH=3-4. The time of infusion, 4 h, 40°C. including ultrasound (intensity 2·104W/m2produced for 15 minutes every hour infusion. The extract was filtered, concentrated by ultrafiltration installation and dried in the spray dryer. Next meal was treated with a 1.5% solution of sodium carbonate followed by obtaining the sodium alginate

The obtained dry extract of fucus contains neutral monosaccharides - fucose 41%, xylose 2,4%, mannose 3,1%, galak the threat of 4.5%, glucose 2,2%, uronic acids of 3.4%, polyphenols 18% and sulphate 21%.

Example 2.

Dry powdered thallus of fucus vesiculosus were extracted by azeotrope of methylene chloride - ethyl alcohol to conventional Soxhlet extractions 6 o'clock Extract was concentrated, dried. Meal algae were extracted by the method of percolation with ethanol 85% at a temperature of 65°C, time of infusion, 5 o'clock Extract was filtered, concentrated in vacuo on a rotary evaporator. Was led at 0÷+4 C for 24 h Mannitol was filtered on a suction filter and dried, purified by the method of column chromatography using alumina. Meal algae were extracted by percolation method with ultrasonic treatment 15%aqueous ethanol, pH=3-4. Time insisting 6 h, temperature 30°C. including ultrasound (intensity 2·104W/m2produced for 20 minutes every hour infusion. The extract was filtered, concentrated by ultrafiltration installation and dried in the spray dryer. Next meal was treated with a 1.5% solution of sodium carbonate followed by obtaining the sodium alginate

The obtained dry extract of fucus contains neutral monosaccharides - fucose 43%, xylose 2.1%, mannose 3,5%, galaksi 4.2%, glucose 2.0%) and uronic acids of 3.1%, polyphenols 20% and sulphate 25%.

The new product is polysacharide-polyphenol complex, which is output differs from polysaccharide complex prototype chemical composition, technological properties and pharmacological effects (anticoagulant) (see tables 1 and 2).

To assess the technological properties of militia were investigated size distribution, flowability, bulk density, residual moisture of the sample extract. Fractional composition of dry extracts of fucus obtained by the present method and the prototype presented in figure 3 and 4. The data show a significant dependence of the fractional composition from the method of preparation and drying of the extract. Lyophilized extract fine, the content of pulverulent fraction is 80,21%. Extract, dried in the spray dryer has a greater number of larger particles from 0,315 to 0,100 mm (81%), fewer pulverulent fraction of 8.1%. Technological properties of militia, obtained by the present method and the prototype shown in the table (see figure 5). Thus, the new dry extract of fucus through the use of spray drying consists of fractions with larger particles, which makes it more manufacturable product, less dust-raising, which is very important for production operations, batching, transporting, filling and packaging.

Tests were carried out pharmacological activity of the extract.

1. Acute toxicity.

Determination of acute toxicity of the extract is carried out in AK is Radicofani Testing laboratory Central Institute of traumatology and orthopedics named. R.R. Vreden method Cerberus (white, 1963) taking into account the recommendations of the pharmacological Committee for the study of General toxic effects of substances (Manual on experimental (preclinical) study of new pharmacological substances. Edited Fisenko FP Ed. The Ministry of health of the Russian Federation. 2000. 400 S.)

Dry extract of fucus (SEF) in a 1% aqueous solution is administered orally to white male mice weighing 18-20 givati was in vivarium conditions with free access to water. Administration of the extract in a 1% aqueous solution was carried out a probe into the stomach on an empty stomach at doses of 250 mg/kg; 625 mg/kg 1250 mg/kg 2500 mg/kg Period of observation of animals - 2 weeks. Calculation of LD50the Cerberus was performed according to the formula:

,

where LD50the dose of the substance that causes death in 50% of animals;

LD100the dose of the substance that causes the death of an entire group of animals;

d is the interval between each two complex doses;

z - arithmetic average of the number of animals in which the effect is observed under the influence of two adjacent doses;

m is the number of animals in each group.

Death is not marked, and clinical signs of intoxication: the General condition, behavior, condition of coat, eating poop in the experimental group did not differ from intact. The autopsy found that nutrena bodies, regional lymph nodes in experimental mice within the physiological norm, the weights of the internal organs in the experimental mice did not show a statistically significant difference from those of intact animals.

Conclusion: the tested dry extract of fucus belongs to class IV, low-toxic substances according to GOST 12.1.007-76 (LD50when oral administration is >2500 mg/kg body weight).

2. Heparinate action - application in the form of ointment. The claimed ointment anticoagulant, as known, includes active and auxiliary substances, which use purified water, emulsifier and preservatives. As the active substance use dry extract of fucus according to claim 2, obtained by the method according to claim 1. The following are examples of manufactured ointments, including stated in 3 independent claim quantitative composition of components with different amounts of ingredients. Example. 1.

Ointment anticoagulant contains the following components, wt.%:

dry extract of fucus - 10;

lanolin - 10;

olive oil - 10;

TWIN 80 - 1;

preservatives - 1;

purified water - the rest (the necessary amount to obtain 100 g of finished product).

Example 2.

Ointment anticoagulant contains the following components, wt.%:

dry extract of fucus - 20;

lanolin - 5;

olive oil - 20;

TWIN 80 - 3;

preservatives - 1,5;

purified water - the rest (the necessary amount to obtain 100 g of finished product).

Example 3.

The ointment contains the following ratio of components, wt.%:

dry extract of fucus - 30;

lanolin - 1;

olive oil - 30;

TWIN 80 - 5;

preservatives - 1,5;

purified water - the rest (the necessary amount to obtain 100 g of finished product).

Example 4.

The ointment contains the following ratio of components, wt.%:

dry extract of fucus - 15;

lanolin - 10;

olive oil - 10;

TWIN 80 - 1;

preservatives - 1;

purified water - the rest.

All of the examples of execution ointments have antikoagulyantny action.

The study of anticoagulant (heparinate) actions ointments containing as main active substance dry extract of fucus obtained by the present method, in comparison with the registered medicinal product "heparin ointment" conducted by CJSC "Saint-Petersburg Institute of pharmacy".

The objects of study were: fucus extract No. 1 (the present method), fucus extract No. 2 (according to the method of the prototype and the product comparison - heparin ointment. The aim of this work was to identify heparinate activity of the investigated extracts No. 1 and No. 2 in the middle and double the th therapeutic doses and the comparison drug for cutaneous application within 7 days (medical scheme).

The study drugs were applied to the pre-released skin area. Animals were determined by prothrombin time (PT) and activated partial thrombin time (APTT) (baseline, after 7 days the introduction). Also animals were determined weight for the correction of doses of study drug. During the experiment it was found that the extract No. 1 in the middle and double therapeutic doses elongation PV and APTT (moderate resorptive action) is comparable with the effect of the comparison drug. Side effects such as hyperemia and dot hemorrhages in the use of the investigational ointments were noted.

Fucus extract No. 1

The sample is provided in a glass jar. The sample represents a soft dosage form of a dark brown color, thick consistency with a characteristic smell of seaweed, soluble in water. The drug composition: 1 g ointment contains:

The active substance - fucus extract No. 1 to 0.15,

Other ingredients: lanolin, olive oil, purified water, emulsifier, preservatives.

Fucus extract No. 2

The sample is provided in a glass jar. The sample is a soft dosage form of a light brown color, thick consistency with a characteristic smell of seaweed, soluble in water. The drug composition: 1 g ointment contains:

The active substance - extras the CT fucus No. 2 - 0,15,

Other ingredients: lanolin, olive oil, purified water, emulsifier, preservatives

Comparison tool: "Heparin ointment". The drug composition: 1 g ointment contains: Active ingredients: heparin sodium - ME 100, benzocaine (benzocaine) - 0.04 g, benzylsuccinic - 0,0008,

Other ingredients: glycerin (glycerol); vaseline; stearin cosmetic "D"; peach butter; emulsifier No. 1; Lanette®; nipagin (methylhydroxybenzoate); nipazol (propylhydroxybenzoate); purified water.

Results studies

PV and APTT was measured in the 0-th and 7-th day of the experiment, blood was taken from tail vein in Eppendorf. The results are presented in tables (see Fig.6 and 7).

The obtained data show a slight lengthening of the APTT in response to the application of heparin ointment in comparison with the baseline 1.3 times. Such a moderate impact on this indicator as observed in clinical practice is admissible, and may be a symptom of minor resorption.

Fucus extract No. 1 in average therapeutic dose showed the same results as the reference product, extending APTT compared to the baseline of 1.3 times, and on the 7th day of the study had a statistically significant difference from the intact group for this indicator.

Fucus extract No. 2 did not have any action on the analyzed parameter.

is C the table shows, that the use of heparin ointment and fucus extract No. 1 in average and double the dose was statistically significantly prolonged prothrombin time in comparison with PV in the intact group. And compared to the base level of ROS in these groups increased in 1.4 times. That may indicate the presence of effects on hemostasis of the studied extract of fucus No. 1.

Fucus extract No. 2 in both doses affect PV slightly, as the tendency to lengthen it.

Conclusions:

Fucus extract No. 2 in the medium therapeutic doses influenced the lengthening of time in the form of trends.

Fucus extract No. 1 in average, and in therapeutic doses were statistically significantly increased the performance of both tests, acting similarly to heparin ointment.

According to the data obtained, namely, time of coagulation (PT, APTT), in intact animals, there was no change in performance, which confirms the adequacy of the methodology. Thus, fucus extract No. 1 in the studied doses (15% and 30%) promising and recommended for further advanced preclinical and clinical studies.

1. A method of obtaining a dry extract of fucus possessing anticoagulant effect by complex processing of fucus vesiculosus, which consists in the fact that the dry thallus algae are crushed, extracted with a mixture of methylene chloride with the alcohol is m ethanol in the ratio was 94.2:5,86% to conventional Soxhlet extractions, concentrated in vacuo on a rotary evaporator, dried in vacuum on a rotary evaporator, dried in a vacuum drying Cabinet with receiving lipid-pigment complex - thick extract, then the meal is extracted by percolation method 85-90%ethanol with forced circulation of the extractant every hour infusion for 3-5 h, filtered, concentrated in vacuo on a rotary evaporator, crystallized, filtered on a suction filter, dried and purified by the method of column chromatography using alumina and receive mannitol cleared, then the remaining meal is extracted by the method of percolation 5-15%aqueous ethanol, concentrate, dried and receive polysaccharide complex - dry extract of fucus, then the meal is extracted with a 1.5%solution of sodium carbonate is treated with sulfuric acid, concentrated on a rotary evaporator, and purified by dialysis through a cellophane membrane is dried in a vacuum drying Cabinet or by lyophilization and get the polysaccharide alginate sodium, characterized in that to obtain a dry extract of fucus algae meal is extracted with 4-6 hours at a temperature of 30-40°C and pH 3-4 by percolation method with ultrasonic treatment, which is carried attached to the outside of percolator source of ultrasound, with the inclusion of ultrasound produced for 15-20 min every hour of nastava the Iya, then the extract is filtered, concentrated by ultrafiltration installation and dried in the spray dryer.

2. Dry extract of fucus possessing anticoagulant effect obtained by the method according to claim 1, which represents a polysaccharide complex, containing fucoidan following composition: neutral monosaccharides - fucose 41-44%, xylose 2-2,5%, mannose 3-3,5%, kalakota 4-4,5%, glucose 2%, uronic acids 3-3,5%; and polyphenols 18-20% and sulfates 21-25%.

3. Ointment anticoagulant comprising dry extract of fucus according to claim 2, lanolin, olive oil, TWEEN 80, preservative, purified water in the following ratio, wt.%:

dry extract of fucus0,15-30
lanolin1-10
olive oil10-30
TWEEN 801-5
preservatives1-1,5
purified waterrest

4. The ointment according to claim 3, characterized in that it contains the following ratio of components, wt.%:

td align="right"> 0,15
dry extract of fucus
lanolin10
olive oil10
TWEEN 801
preservatives1
purified waterrest



 

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7 dwg, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely a method for preparing an agent possessing immunomodulatory activity. The method for preparing an agent possessing immunomodulatory activity characterized by the fact that herbal composition containing common motherwort herb, knotgrass herb, pot marigold blossom, licorice rhizome and root, Alexandrian laurel rhizome and root, rosehip, schizandra fruit, linseeds, sequentially twice extracted in 60-70% ethanol, twice in 40-50% ethanol, and once in purified water under specific conditions; then the aqueous-alcoholic extracts are concentrated in vacuum; the aqueous still residues are combined with the aqueous extract, then filtered, boiled out, purified by separation, boiled out additionally, dried in a vacuum dryer and further milled.

EFFECT: method enables preparing the agent possessing manifested immunomodulatory activity with the high content of active substances.

13 tbl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely a method for preparing an agent possessing tonic activity. The method for preparing the agent of true ginseng roots, possessing tonic activity by three extractions of the ground raw material in 40-80% ethanol with using three extraction vessels in the specific environment; at first the extraction process takes place at room temperature for 12 hours, while the third extraction is conducted at temperature 90°C for 30 minutes in each extraction vessel.

EFFECT: method enables preparing the high-saponin agent possessing manifested tonic activity.

3 dwg, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to pharmaceutical industry and deals with creation of medication of vegetable origin based on biologically active substances of Linaria vulgaris, which has diuretic and anticoagulation activity. Method of obtaining medication, which has diuretic and anticoagulation activity, consists in the following: extraction of milled grass of Linaria vulgaris with ethyl alcohol is carried out with successive drawing with 3 parts of extractant by method of remaceration with heating and connecting vacuum at the stage of discharge, under specified conditions.

EFFECT: method makes it possible to obtain medication, which has high diuretic and anticoagulation activity.

3 tbl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to pharmaceutical industry, in particular to method of obtaining immunotropic preparation. Method of obtaining immunotropic preparation by extraction of air-dry leaves of Lophanthus anisatus Benth with aqueous ethyl alcohol, heating, cooling, filtering, solvent evaporation, residue dissolving and extraction with chloroform, filtering and drying of sediment, under specified conditions.

EFFECT: method makes it possible to obtain efficient immunotropic medication.

3 tbl, 5 ex

FIELD: medicine.

SUBSTANCE: invention refers to cosmetic and pharmaceutical industry, particularly to a method for preparing a water-in-oil extract of herbs and/or flowers. The method for preparing a water-in-oil extract of herbs and/or flowers involving pre-watering or moistening of the herbal raw material in deionised water, and further extraction of the wet or swollen raw material by single-flow percolation or static mixing with an extractant presented by a water-in-oil emulsion containing oil phase 3-15 wt % specified in a group consisting of natural milk, curdy whey or a mixture thereof or a mixture thereof with milk whey under certain conditions.

EFFECT: method enables preparing the extract with the high content of biologically active substances while being highly effective and ecologically-friendly.

5 cl, 10 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely to a method for preparing a chromogenic complex of artist's conk. A method for preparing the chromogenic complex of artist's conk involving the staged extraction of a ground fruiting body of artist's conk in an aqueous solution of sodium hydroxide, herewith the first-stage extract and pulp are prepared at first; then the pulp is extracted to prepare the second-stage extract; the first-stage and second-stage extracts are combined, and the chromogenic complex is deposited by adding hydrochloric acid to the combined extract under certain conditions.

EFFECT: method enables providing higher yield of the end product having higher antioxidant activity.

1 tbl, 4 ex

FIELD: biotechnologies.

SUBSTANCE: invention refers to cyclohexyl ammonium salt of 2-[3-methyl-7-(1,1-dioxotiethanyl-3)-1-ethyl xanthenyl-8-tio]acetic acid of the following formula: .

EFFECT: obtaining a new compound that shows antithromboembolic action and can be used in medicine.

2 cl, 2 tbl, 3 ex

FIELD: biotechnologies.

SUBSTANCE: invention refers to derivatives of oxazolopyrimidine in any of their stereoisomeric forms, or in the form of a mixture of stereoisomeric forms specified in Claim 1.

EFFECT: oxazolopyrimidine derivatives having agonistic activity in relation to Edg-1 receptor.

5 tbl, 319 ex

FIELD: chemistry.

SUBSTANCE: invention relates to N-carb(glutaminyl)oxymethylimidazo[4,5-e]benzo[1,2-c; 3,4-c']difuroxane of formula .

EFFECT: obtaining a novel compound which can be used as a medicinal preparation which inhibits thrombocyte aggregation.

1 dwg, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula (I) and/or stereoisomeric forms thereof and/or mixtures of said forms in any ratio and/or a physiologically tolerant salt of the compound of formula , where: X denotes -C(O)- or -SO2-, U denotes an oxygen atom or -(C0-C4)alkylene, A denotes an oxygen atom, -C(O)-NH-, -NH-C(O)- or -(C0-C4)alkylene, V denotes: 1) -(C2-C9)alkylene, where alkylene is unsubstituted or mono-, di- or tri-substituted, independently of each other, by an -OH group, 2) -(C3-C9)alkenylene, D denotes -(C1-C2)alkylene, Y denotes: 1) a covalent bond, 2) -(C6-C14)arylene-, or 3) Het, where Het denotes pyridyl or imidazolyl, R1 denotes: 1) a hydrogen atom, 2) -(C1-C6)alkyl, R3 denotes: 1) -(C2-C6)alkylene-NH2, 2) -(C1-C4)alkylene-SO2-(C1-C4) alkylene-NH2 or 3) -(C0-C4) alkylene-Het, where Het denotes pyridyl or piperidyl, where Het is unsubstituted or substituted with -NH2, R6 denotes: 1) a hydrogen atom, 2) -(C1-C6) alkyl, where the alkyl is unsubstituted or substituted, independently of each other, by a R16 group, 3) -(C0-C4) alkylene-Het, where Het denotes pyridyl, where -(C0-C4) alkylene and Het are unsubstituted or substituted, independently of each other, by a R16 group, 4) -(C0-C4) alkylene-phenyl, where -(C0-C4) alkylene and phenyl are unsubstituted or substituted, independently of each other, by a R16 group, or 5) -(C0-C4) alkylene-(C3-C8)cycloalkyl, R7 denotes a hydrogen atom, halogen or -(C1-C6)alkyl, R8 denotes a hydrogen atom or -(C1-C6)alkyl, R9 denotes a hydrogen atom, and R16 denotes -NH2, which are inhibitors of the active thrombin-activated fibrinolysis inhibitor, as well as a method for production thereof, a medicinal agent based thereon and use for prevention, secondary prevention and treatment of one or more disorders associated with thrombosis, embolism, hypercoagulation or fibrosis changes.

EFFECT: improved properties of compounds.

7 cl, 1 tbl, 9 ex

FIELD: chemistry.

SUBSTANCE: described are novel triazolopyridazines of general formula , stereoisomeric or tautomeric forms thereof and physiologically acceptable salts thereof, where Q1 denotes H, -C1-6alkyl, optionally substituted with fluorine, or -C3-6cycloalkyl; Q2 and Q3 independently denote H, -C1-6alkyl; R1-R3 independently denote H, -C1-6 alkyl, -C3-6cycloalkyl, -O-C3-6cycloalkyl, -O-C1-8alkyl, a heterocyclic residue etc; R4-R8 independently denote H, -C1-6 alkyl, -OH, -O-C1-8 alkyl, halogen, SF5 etc, a method for production thereof and use as medicinal agents.

EFFECT: compounds have antithrombotic activity and particularly inhibit the protease-activated receptor.

6 cl, 2 tbl, 242 ex

FIELD: medicine.

SUBSTANCE: present invention refers to medicine, particularly to pharmacology, and describes a method for correction of disturbed functional activity of platelets, consisting in using Melaxen as a corrector for the disaggregation and hyperaggregation state of the platelets to be orally administered into white non-linear mature rats in a dose of 1 mg/kg once a day within the 7-day therapeutic course.

EFFECT: invention aims at extending the range of preparations having an ability to control the aggregation properties of platelets depending on the nature of the haemostatic disorders.

4 tbl, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to anticoagulant compounds of formula (I) : and pharmaceutically acceptable salts thereof, wherein R represents C1-C8 alkyl substituted by at least one halogen specified in chlorine or fluorine.

EFFECT: invention refers to pharmaceutical compositions containing the compounds and salts thereof, as well as to using the compounds and salts thereof for treating bleeding disorders, to a method of treating, inhibiting epoxyredutase, vitamin K and coagulation factor synthesis.

31 cl, 3 tbl, 11 ex, 10 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: there are described new triazolium salts of general formula (I) stereo- and tautomer forms and physiologically acceptable salts thereof, wherein X- C-R1 or N, R1-hydrogen, C1-6alkyl or halogen; A- is an anion of a pharmacologically acceptable organic or inorganic acid; Q1 is hydrogn, -C1-6alkyl optionally substituted, -C3-6cycloalkyl, -C(O)-O-R11 or - C(O)-R11; R11 -C1-6alkyl; Q2 and Q3 are hydrogen; R2- R9 independently mean hydrogen, -C1-6alkyl optionally substituted, -O-(C1-8)alkyl optionally substituted, etc., and using the above compounds as a drug preparation.

EFFECT: compounds possess antithrombotic activity, particularly, they inhibit the protease-activated receptor 1 (PAR1), and may be used in treating the diseases such as myocardial infarction, angina, stroke, and others.

4 cl, 2 tbl, 86 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a solid oral pharmaceutical formulation containing a hydrolised form of 5-chloro-M-({(5S)-2-oxo-3-[4-(3-oxo-4-morpholinyl)phenyl]-1,3-oxazolidin-5-yl}methyl)-2-thiophenecarboxamide in the amount of 1 to 60%. The formulation contains sodium lauryl sulphate as a wetting agent and hydroxypropylmethyl cellulose as a hydrophilic binding agent. The pharmaceutical formulation is presented in the form of a tablet.

EFFECT: formulation according to the invention provides higher biological availability of the active ingredient.

17 cl, 2 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a biologically active substance possessing the antiaggregant properties. The technical effect: what is produced is a drug preparation of a new compound of (2E)-3-[1-(2-hydroxy-3-piperidin-1-ylpropyl)-1 H-indol-3-yl]-1-(2-thienyl)prop-2-en-1-one hydrochloride of formula I possessing antiaggregant action:

.

EFFECT: substance may be used in medicine for producing a drug preparation for preventing the conditions associated with a high thrombogenic potential of blood.

1 cl, 2 tbl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, namely to pharmacy and concerns the development of an ointment containing mechanically activated amorphous or amorphous crystalline calcium gluconate, and may be used in the integrated treatment of skin diseases related to calcium deficiency, namely to the integrated treatment of dermatoses: psoriasis, atopic dermatitis, eczema and keratoderma. As an active ingredient, the ointment contains mechanically activated amorphous or amorphous crystalline calcium gluconate; an ointment base is Vaseline, glycerol, emulsifier No.1, Lutrol F 127 and purified water in the following proportions, wt %: mechanically activated amorphous or amorphous crystalline calcium gluconate-5.0; Vaselin-10.0; glycerol-10.0; emulsifier No.1-8.0; lutrol F 127-2.0 and purified water up to 100.0.

EFFECT: ointment possesses high therapeutic activity, provides the uniform release of the active ingredient that is mechanically activated amorphous or amorphous crystalline calcium gluconate, the manifested prolonged action and applicability by the patient.

4 ex

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