Method of obtaining medication, possessing antioxidant, cholagogic, hepato radio- and nephroprotective, antiradical, wound-healing activity

FIELD: medicine.

SUBSTANCE: wastes of confectionery industry production, milled husk of cacao beans, are drawn in water at temperature 20°C for 2 hours with following extraction by hot water (80-90°C) in battery of four extractors, thickening of extract and drying are carried out.

EFFECT: extension of medication activity spectrum and decrease of power consumption.

8 tbl

 

The invention relates to the pharmaceutical industry, namely the production of therapeutic and prophylactic drugs from edible raw materials.

The technical result of the invention is to develop a method of obtaining funds simultaneously possessing antioxidative, choleretic, hepato-, radio - and nephroprotective, antiradical, wound-healing effect.

A method of obtaining from the husk of the cocoa beans, cocoa-Vell means having hepatoprotective, choleretic, antioxidant and radioprotective action, which is as follows: 50 kg cocoa-Vell is extracted with 250 kg of hot water (80-90°C), water extraction will shuchat, dried, crushed and receive 11,2 kg light brown amorphous powder with a yield of 22.4% [1].

A significant disadvantage of the known method are significant energy costs at the stage of condensation of the extract, since the total amount warded off the water is up to 750 kg per 33,6 kg of the desired product.

The aim of the invention is the extension of the spectrum of pharmacological activity of the target product, simultaneously possessing antioxidative, choleretic, hepato-, radio - and nephroprotective, antiradical, wound healing effect and increase the yield of the target product, reducing energy and material costs stadiongasse aqueous extracts of cocoa-Vell.

This objective is achieved in that the extraction process cocoa-Vell is carried out in a battery of four percolation water method remuneratio with mandatory infusion with cold water, it is illustrated by the following example of a specific implementation.

In the battery of four extractors, equipped with a steam jacket and a stirrer, a load of 10 kg cocoa-Vell and 70 kg of water (20°C), mix and leave for 2 hours to swell. Then in the first extractor add 100 kg of hot (90°C) of water, heat the entire mass to 90°C and under stirring to keep the temperature from 80 to 90°C for three hours. Water extraction from the first extractor is drained and transferred to the second extractor with swollen raw material, heated to 90°C. and under stirring incubated three hours at 80-90°C. In the first extractor pour 100 kg of hot (90°C) of water, and with stirring, keep the temperature from 80 to 90°C. Similar operations are performed in all four extractors. From the fourth extractor as obtain the saturated extract. Using the fifth extractor does not lead to a significant increase in the content of extractives (table 1).

Table 1
Room extractor The content of extractives in extracts from different extractors, kg/%
12345
12,144/21,44
21,163/11,162,723/27,23
30,306/3,063,635/36,35
40,264/2,644,482/44,82
50,132/1,324,486/44,86

Extract from the fourth extractor concentrated to dryness and obtain the target product with the release of 4,482 kg (44,82%). The first extractor becomes the fourth. The process learned what I continue as described above to produce the desired amount of the target product.

The proposed method for obtaining the target product in contrast to the known widen the range of pharmacological activity of the target product, to increase its output in 2 times and significantly (5-6 times) to reduce the number chamaemoro extract (instead of 500-600 kg is just 100 kg)that will significantly reduce energy and material costs.

The study choleretic activity conducted by the quantity of excreted bile. Animals of the control group were injected water. As the comparison drug used official cholagogue "Flamin". To identify the hepatoprotective activity was determined hepatoprotective index in rats to carbon tetrachloride in a toxic dose.

At the same time was determined by a percentage reduction of the enzyme alanine aminotransferase (Alat). As the comparison drug used official hepatoprotective means "Zelibor" (table 2).

Antioxidant activity was determined in accordance with the method of Reference to reduce malonic dialdehyde (MDA). Drug comparison served as vitamin E (table 2).

Table 2
MedicationQuantity of ratsCholeretic the activity, % increaseQuantity of ratsHepatoprotective activity % reduction AlatQuantity of ratsAntioxidant activity, % reduction of MDA
Target product (Cavegol)1094,71023,31-043,5
The placeholder1095,51023,81044,4
Flamin1053,8
Zelibor1051,0
1029,5

In table 3 the results of studies of wound healing ability after radiation damage to the skin.

Table 3
MedicationQuantity of ratsReduction of terms of epithelialization, days /%The starting dates of wool regrowthTermination of the wool regrowth
Control (nat. R-R)2041,0±1,5/10050,3±2,059,4±2,5
The placeholder2022,5±1,5/54,8828,4±2,034,2±2,5
Target product2021,8±1,5/53,1728,5±2,034,0±2,5
Atony2024,8±1,5/60,4931,0±2,540,3±2,5

Are given in table 2 and 3 the results of the research show that the target product (cavegol)obtained by the proposed the method, in comparison with the prototype almost does not reduce choleretic, hepatoprotective, antioxidant and wound healing (by radiation damage to the skin) activity.

Hepato - and nephroprotective activity of the target product was studied on a model of chronic endotoxemia (HAT) in rats induced by carbon tetrachloride (5 ml/kg 5 times a week), and on the sixth day of the additionally injected intraperitoneally bacterial lipopolysaccharide (Sal. typhimurium, Sigma, USA at a dose of 0.2 ml/kg) [2]. Mortality of animals for 30 days was 10%to 90 days - 40-50%. In the comparison group (norm) used intact animals. The experimental group of animals at 7 days was received additional cavegol at a dose of 25 mg/kg orally. As the comparison drug use Essentiale, which was administered intraperitoneally in the form of a solution of 0.1 ml/kg once a day for background modeling the HAT.

In the liver histomorphometry was calculated volume fraction (ML, mm3)occupied by connective tissue, hepatocytes (%) and ML lipids, necrosis of hepatocytes (%).

In table 4 the results of morphological studies of liver tissue. It is established that the development of the HAT is accompanied by a progressive degeneration and necrosis of hepatocytes with the reaction of the microcirculation, macrophages and lymphocytes on them. Changes perisinusoidal cells and vascular reconstruction sposob the job for keeping the liver moderately expressed interstitial cytolysis, and by the 30th day revealed signs of hepatovirus. Morphometry of the experimental group of animals treated with the target product (cavegol)showed a significant limitation of all morphometric appearances secondary liver damage when CHET (table 4). Course introduction Essentiale led to partial restriction reduce the OD of hepatocytes and increase the proportion of necrotic hepatocytes, characteristic of chronic FL. At the same time increasing the OD of the connective tissue was also delayed compared with the control (table 4). Target product (cavegol) caused similar effects, comparable with Essentiale on the terms 7 and 14 days. On the 30th day of its effect on the conservation pool of hepatocytes was higher than that of Essentiale and in relation to the slow growth of connective tissue is slightly lower, but comparable with Essentiale.

Table 4
The influence of the studied drugs on the performance of the morphometry of liver, with HAT in rats
The experimental groupThe timing of the experiment
7 days21 days30 days
OD connective tissue, % (normal is 1,8±0,5)
Control6,4±0,59,3±0,710,2±0,8
Target product (cavegol)3,0±0,3*5,4±0,6*6,4±0,6*
Essentiale3,1±0,4*5,2±0,7*6,2±0,8*
OD hepatocytes, % (norm - 63,5±2,9)
Control58,0±3,451,4±3,048,0±2,9
Target product (cavegol)63,1±3,759,2±3,7*58,0±3,3*
Essentiale61,5±2,958,0±3,1*51.4±3,0*
Necrosis of hepatocytes, % (norm - 4,5±0,5)
Control18,2±1,422,4±1,720,6±1,6
Target product (cavegol)14,0±1,1 12,5±0,8*13,8±0,9*
Essentiale12,5±1,110,3±1,0*17,1±1,4*
OD lipids, % (norm -3,8±0,4)
Control9,2±1,415,0±1,911,4±1,2
Target product (cavegol)6,0±0,7*8,4±1,1*9,0±1,0
Essentiale6,6±0,7*12,2±1,5*10,5±1,1
* Significant differences from control, p≤0,05

When assigning animals Essentiale manifestations dysmetabolic nephropathy, characteristic of chronic endotoxemia, somewhat smoothed out. The formation of nephrosclerosis was not accompanied by violations vnutripochechnykh urodynamics, the phenomena of degeneration and necrosis of the epithelium of the tubules were not as striking (table 5).

When the target product, unlike Essentiale, has been a slowing down of growth of ML connective tissue and the percentage of scleratinian glomeruli, as well as reducing the urinary space in KL the barrels. Keeping the size of the nuclei of the epithelium of the tubules, in contrast to their reduction when CHET was equally characteristic of Essentiale and kaihola.

Table 5
The influence of catechol and Essentiale on indicators of kidney morphometry with HAT in rats
The experimental groupThe timing of the experiment
7 days21 days30 days
OD connective tissue, % (rate of 7.3±0,7)
Control12,7±0,713,7±0,815,5±1,1
Target product (cavegol)8,9±0,6*9,1±0,8*8,4±0,7*
Essentiale12,0±0,712,2±1,015,0±0,8*
Urinary space of glomeruli (norm - 1,8±0,2)
Control1,4±0,21,1±0,4 0,9±0,2
Target product (cavegol)1,9±0,21,7±0,2*1,6±0,3*
Essentiale1,8±0,21,1±0,2*1,0±0,3*
WITH the epithelium of the tubules, μm3(norm - to 102.9±4,9)
Control104,0±9,885,9±7,0of 82.8±6,1
Target product (cavegol)110,4±10,1to 102.3±9,7the 98.9±7,8
Essentialeof 101.4±9,2of 100.2±9,596,4±7,7
Sclerotinia the glomeruli, % (normal - no)
Control01,0±0,38,0±0,8
Target product (cavegol)004,4±0,4*
Essentiale00 3,9±0,3*
* Significant differences from control, p≤0,05

Thus, when using kauhola able to partially restrict cytotoxic, vascular and fibroplasias mechanisms of tissue damage liver and kidneys, peculiar HAT. Its protective effects against liver comparable to the reference product, and in relation to kidney - exceed action Essentiale.

Antiradical activity (ARA) of the target product was determined by the degree of quenching of the chemiluminescence in the system, generating free radicals, using chemiluminometer HLM 1TS-01 [3]. When you add into the system, generating free radicals, solution of the target product in volume (V) from 1 to 30 ál can determine the degree of quenching of the chemiluminescence. The initial concentration of 1 mg/ml, the solvent used water. The amount of quenching of chemiluminescence as a percentage of control X calculated according to the formula X=100%-(·100%)/K, where a is the value of damping chemiluminescence solution of a substance To suppress the chemiluminescence of solvent.

Cavegol at a concentration of 0.24 mg/ml leads to the quenching of chemiluminescence at 42.5%.

As the comparison drug used well-known antioxidant BHT (4-hydroxy-3,5-di-tert-butyl-1-methylbenzol), which is described and antiradical the I activity [4]. Research ARA dibunola conducted under the same conditions showed no quenching of free radicals even at a concentration of 1 mg/ml

Thus, the magnitude of inhibition of chemiluminescence target product significantly exceeds the activity dibunola.

Wound healing activity of the target product burns is set to the 12 laboratory rats of both sexes at the age of 80 days. Results of experiment was used a comprehensive criterion for determining the effectiveness of the healing of burns in experimental animals.

As a first criterion was performed histological examination of burn surfaces. Sections of skin obtained at 3, 7 and 14 days of experiment. The skin samples were placed in 10% formalin solution to the posting by the standard technique and colouring with hematoxylin and eosin. This study assesses the impact of therapy on the repair processes.

The following criterion data bacteriological research in order to objectively assess the dynamics of wound healing by identifying the presence of microflora in the composition of wound and holding its qualitative assessment. The material for bacteriological analysis was 56 swabs to burn the surface of the skin of rats. As control was used 8 swabs from the burn surfaces, not p is Girgashite treatment.

Quantitative and species composition of the microflora of burn surface studied by bacteriological methods by planting swabs from the surface of the burn on differential diagnostic environment to determine the number of certain microorganisms after application of the target product and the comparison drug "Levomicol".

In the study of microbiocenosis of burn surface was taken into account: 1) the total number of microorganisms; 2) the microbes Enterobacteriaceae; 3) Staphylococcus; 4) the fungi of the genus Candida. Estimated frequency of occurrence of individual species of microorganisms depending on the biotope and products used for treatment of burns, the density of bacterial contamination in the dynamics on the 7th and 14th day of treatment of burn surface with the index of dominance, defined by the formula of Odum Yu [5].

The final criterion was to evaluate the dynamics of the size reduction of a skin defect: the area of the wound in percent, based on the beginning of treatment - 100% burn surface (table 6).

td align="center"> Target product (cavegol)
Table 6
The area of skin wounds (% of burn area)
Investigational drugs3 days7 days14 days
85,355,626,6
Levomekol (Rev. comparison)94,562,130,8
Control (without treatment)90,0*45,836,9
* In the early stages of animals that did not receive treatment, the surface area of the wound is reduced by 10-15%

Moreover burn surface viewed from the position of dominance of each species as an indicator using the frequency of occurrence [6]. It was allocated persistent species (prevalence > 50%), additional (25-30%) and casual (less than 25%).

Analysis of the microflora, allowing to divide it into a permanent and more, presented in table 7.

Table 7
The frequency of occurrence of ecological groups of microorganisms in the microbiocenosis of burn surfaces rats
No. biotopeEnvironmental groups, %
StaphilococcusStreptococcusEnterobacteriaceaeFungi of the genus Candida
Target product (kaweka)62,525,075,025,0
Levomekol87,525,062,537,5
Control100,037,587,575,0

The main representatives sown with the surface layer of burn surface, are staphylococci. Thus, staphylococci are the dominant representatives of burn surface, with the largest share among Staphylococcus belongs to the tendency of development, S.epidermidis, S.aureus (table 8).

Table 8
The degree of dominance of species of staphylococci in the community structure of burn surface
No. biotopeSpecies of staphylococci,%
S. saprophyticusS. epidermidisS. aureus
Target product (cavegol)41,842,515,7
Levomekol23,631,345,1
Control3,441,8of 54.8

The analysis of the dominance structure of staphylococci showed that the use of the target product prevailed saprophyte aureus representative of the resident flora. In contrast to burn the surface of the comparison group absolutely dominated Staphylococcus aureus (54,8%) and epidermal Staphylococcus with hemolytic properties. In habitats that are processed product comparison-levomekol, dominated Staphylococcus aureus (45,1%), which complicates the course of wound and burn process and causes of purulent complications.

When studying the total amount of bacteria in the dynamics found that this index was significantly decreased by 14 days of the experiment when using the target product, namely from 131, 5mm±17,1 CFU/cm2- on the 7th day of the experiment, to 100.9±12,3 CFU/cm2/sup> - on the 14th day (p<0,05). The treatment of burn surfaces product comparison levomekol not resulted in significant reduction of total bacterial contamination. The decrease in the density of microbial associations on the 14th day of the experiment was recorded on the burn surfaces that are not treated with drugs. 3 day burn all surfaces were covered with a thick scab, and some of them on day 7, moved in the third phase of healing - epithelialization, formation and reorganization of the scar.

These histological studies show that the healing of burn surface occurred during the application of both drugs, as well as in the control sample not treated with drugs. However, the rate of epithelialization was different. On the 3rd day of the experiment showing signs of epithelialization, as evidenced by basophilic staining micropreparative under a scab. The presence of cell-leukocyte infiltration actively affect the healing time and, accordingly, the rate of epithelialization. Thus, cell infiltration was slightly evident when using the target product and increased when using levamisole (drug comparison), extending the period of epithelialization. On day 7, marked clavate basophilic proliferation aimed at healing a burn on top of the spine, the most pronounced under the influence of the target product. Under the epithelial layer appears immature granulation tissue, with a rich vascularization, and growth of muscle fibers under the basal membrane. It should be noted that the use of the drug levomekol epithelialization occurs with elements of pathological regeneration, manifested in the form of growth of the epithelium around the hair follicle.

14 day registered sawtoothed epithelium with almost complete closure of the burn defect. Under kapitalizirovana plots are observed proliferation of connective tissue. The peculiarity of burn surface when applying the target product is lasting scab (until complete epithelialization), resulting in aseptic management of wounds and explains the low level of bacteriological contamination.

Thus, evaluating the degree of healing of burn wounds, you can come to the conclusion that most early epithelialization and closure of burn defect occur when using the target product (kaihola), the area of the burn and the rate of epithelialization was higher and was accompanied by the absence of purulent complications.

Thus, the inventive method allows to obtain the target product with both antioxidative, choleretic, hepato-, radio - and nephrops tectorum. antiradical activity, wound healing activity, as well as to improve the yield of the target product, to reduce energy and material costs at the stage of condensation of aqueous extracts of cocoa-Vell.

Literature

1. RF patent №2033176, CL AC 35/78.

2. Novochadov NR. Actual problems of morphology. - Tr. Siberian honey. University, Tomsk, 2002, s-106.

3. Pashkov A.N. Clinical laboratory diagnostics. M: Medicine. 5-6, 1992, pp.62-63.

4. Mashkovsky PPM Medicines. M., Ed. 14-E. LLC "New Wave", 2004.

5. Odum Yu Ecology: TRANS. from English. - M.: Mir, 1986. - 376 S.

6. Sytnik SR Environmental characteristics of the community // IMEI. - 1990. No. 9. - P.27-31.

The method of obtaining funds, possessing antioxidative, choleretic, hepato-, radio-, nephroprotective, antiradical and wound-healing effect, by infusion in water at 20°C for 2 h, followed by extraction with hot water (80-90°C) waste production confectionery industry - shredded husks of cocoa beans in the battery of four extractors, thickening of the extract and drying.



 

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16 cl, 1 tbl, 1 dwg, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention concerns medicine. A soft drink on the basis of herbs consists of the concentrated vegetative extract obtained from an admixture of herbs, chosen of Sida sps., Boerhaavia diffusa, Vitis vinifera, Tinospora cordifolia and Withania somnifera, along with green palm sugar, the agent causing a fermentation, and carbonised water, in which proportion Sida sps.: Boerhaavia diffusa: Vitis vinifera: Tinospora cordifolia: Withania somnifera in a powdery admixture makes (15-20 : (5-10 : (15-20 : (5-10 : (5-10); Proportion of wt/wt: green palm sugar: the concentrated vegetative extract makes from 1 : 3 to 1 : 4; wt/wt, a proportion of carbonised water: the admixture of the concentrated vegetative extract, green palm sugar and the agent causing a fermentation, makes from 1 : 3 to 1 : 5. Perform: (a) obtaining of parts of Sida sps., Boerhaavia diffusa, Vitis vinifera, Tinospora cordifolia and Withania somnifera; (b) crushing of parts of plants and their mixing with obtaining of a powdery admixture; (c) water addition to a powdery admixture from a stage (b) for obtaining of a water extract; (d) concentration of a water extract from the stage (c); (e) filtration of the concentrated extract from the stage (d); (f) admixture of green palm sugar to the filtered extract from the stage (e); (g) addition of Sacromyces strain and the agent causing fermentation, to the admixture from the stage (f); (h) fermentation of the admixture from the stage (g) during the period of time from 3 till 6 days; (i) filtration of the fermented admixture from the stage (h); (j) concentration of the fermented filtrate from the stage (i) with obtaining of the basic solution and (k) mixing of the basic solution (j) with the carbonised water in wt/wt proportion from 1 : 3 to 1 : 5 with obtaining of a soft drink on the basis of herbs.

EFFECT: creation of a soft drink on the basis of herbs.

11 cl, 5 tbl, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention concerns the pharmaceutical, food and cosmetic industry. Perform extraction of the raw material crushed with water with obtaining of a water extract and pulp. The pulp, obtained after extraction of raw materials by the water, is extracted in two steps with ethyl alcohol. At the first step of extraction the constrictor is extracted within 4-6 hours using the following proportion: ethyl alcohol 1:(5-7) with concentration of alcohol of 30% either 50%, or 70%, at temperature of 60-75°C, obtaining an ethanolic extract of the first step of extraction and pulp; then, this pulp is repeatedly extracted within 4-6 hours with ethyl alcohol using the following proportion: ethyl alcohol 1:(3-5) with concentration of alcohol of 30% either 50%, or 70%, at temperature of 60-75°C, obtaining an ethanolic extract of the second step of extraction and pulp. After that the ethanolic extract of the first step of extraction is aggregated with the ethanolic extract of the second step of extraction.

EFFECT: increase in active substances content in an extract.

1 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: prevention method of acute respiratory infections is developed for children with frequent recurrent diseases of respiratory tract. It consists in simalteneous administration of arbidol - 1 tablet (50 mg) twice a week during 3-4 weeks combined with selenium-active - 0.5 tablet (25 mcg of selenium) once a day during 3-4 weeks. In the course of 12 months the method helps to reduce frequency and general time duration of respiratory diseases by factor of 1.9 due to simulteneous correction of immune and metabolic state disorders.

EFFECT: reduction of frequency and general time duration of respiratory diseases.

4 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely, to experimental pharmacology. Method includes introduction of actoprotective preparation reamberinum intravenously in doze of 10 mg/kg of animal weight in morning hours 40 minutes before carrying out Porsolt tests and tests of vertical screen-net.

EFFECT: extending arsenal of means for enhancing work capacity of laboratory animals in experiment.

2 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention refers to composition for prevention, symptom relief and/or treatment of skin and hair/animal fur diseases caused by inflammatory reactions, environment hostilities, ageing or cancer. Specifically the present invention concerns applications of flavonone compounds or derivatives in food, cosmetic or pharmaceutical compositions.

EFFECT: improved human or domestic animal skin and animal fur.

16 cl, 1 tbl, 5 ex, 4 dwg

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