Phyto-nutrient steps

 

(57) Abstract:

The invention relates to medicine, namely to funds on the basis of biologically active substances of plant origin, and can be used as a means of providing anti-inflammatory, immunomodulatory, anabolic effects. Phyto-nutrient action contains alfalfa extract (50 - 90 wt.%) and fillers, as fillers calcium gluconate, sucrose, calcium stearate and talc (9,6 - to 53.0 wt.%). The invention provides for the extension of the scope of the drug due to the absence of contraindications and side effects. 9 table.

The invention relates to medicine, namely to funds on the basis of biologically active substances of plant origin, and can be used as a means of providing anti-inflammatory, immunomodulatory, anabolic effects.

The closest analogue of the invention is a biogenic stimulator "Biosed", which is an aqueous extract of fresh grass Sedum great. The drug enhances the metabolism and regeneration, has a tonic and anti-inflammatory action. It is used as ancillary the surgical and therapeutic practice (Mashkovsky M. D. Medicinal product. -M.: Medicine.in 1993.-H. 2.-S. 175). However, "Biosed has side effects, as well as its application can be marked hyperemia and urticaria rash, and he has contraindications (agilia and malignant neoplasms), which limits its scope.

The objective of the invention is the expansion of the means of nutrient action.

The technical result is the extension of the scope of the drug due to the absence of contraindications and side effects.

This technical result is achieved by the fact that the phyto-nutrient action contains as the basis of the condensed extract of alfalfa and as fillers calcium gluconate, sucrose, calcium stearate and talc in the following ratio, wt.%:

The condensed extract of alfalfa (in recalculation on 100% dry matter) - 50-90

Calcium gluconate - 4-25

Sucrose - 5-25

Calcium stearate and 0.2-1

Talc - 0,4-2

Condensed alfalfa extract obtained by treatment of hay - alfalfa grass extractant containing soluble salts of the metals in mg per 1 kg of plant matter, for example, in the following ratio: Mo-8,0; Ba-10,0; Pb-20,0; Co-1,05; V - 1,0; Cr-0,5; Zn is lovenow weight or dry granules with a residual humidity 10 - 15%, of which prepare a 50% solution.

The inventive herbal remedies obtained as follows. 50% solution of condensed alfalfa extract is thoroughly mixed in a porcelain mortar with a powder of calcium gluconate until smooth. Mass is spread in cassettes stainless steel and dried at a temperature (455)oC for 36 hours and Then dried mass is crushed, sieved through a sieve with the hole diameter of 1 mm and mixed with shredded mixture of sucrose, calcium stearate and talc to obtain a homogenous tablet blend. The powder thus obtained, well tabletroute, preserving the activity and properties shown GF XI edition. The finished product is a tablet from coffee to dark brown, bitter coffee taste and a peculiar smell.

Example 1. To obtain 1 kg of tablet mass of phytomedicine in a porcelain mortar mix thoroughly 900,0 g condensed extract of the herb alfalfa and 44.0 g of calcium gluconate to obtain a homogeneous mass and dried at (455)oC for 36 hours and Then the mass is crushed, sieved through a sieve with the hole diameter of 1 mm and mixed with powdered mixture of 50.0 g tabletirujut.

Example 2. Technology of preparation of the drug are similar to those described in example 1. Mixing take 500.0 g of condensed extract of alfalfa; 220,0 g of calcium gluconate; of 250.0 g of sucrose; 10.0 calcium stearate and 20.0 g of talc.

Bashkir gosmeduniversiteta experimental tests of specific and General pharmacological activity of the proposed plant-based.

Anti-inflammatory herbal remedies studied on the model carrageenans inflammation of the paws of rats (first series of experiments) and on the model of implanting a cotton swab in the subcutaneous tissue of rats (II series). I a series of experiments were conducted on three groups of rats for 6 each. Previously all animals was determined concomitance volume feet. Then the animals of the 1st group received inside pills herbal drug in a dose of 50 mg/kg, 2-nd group - ortofen in the dose of 10 mg/kg (U50), and group 3 was intact. Then all animals were caused by the inflammation of the feet subplanetary the introduction of 0.1 ml of 1% suspension carragenine and determined the amount of inflamed paws after 3, 6, 24, 48 and 72 h (table. 1).

A second series of experiments conducted on three groups of rats for 10 each. Under a light Nembutal anesthesia (30 mg/kg) all animals in subcutaneous cleaopatra dose of 50 mg/kg, 2nd group - ortofen in the dose of 10 mg/kg, and 3-I was intact. Then, animals were scored by decapitation, removed cotton swabs, weighed them, and dried in a thermostat at 60oC for 3 days and weighed again. The difference of weight of the "wet" and "dried" tampon judged on the impact of drugs on the exudative phase of inflammation, and by the difference of weight "dried" and the original weight of the tampon judged on the impact of drugs on the proliferative phase of inflammation (table. 2).

Thus, herbal drug has a pronounced anti-inflammatory activity, showing antiflogistic, antiexudative and antiproliferative activity.

To assess the impact of herbal remedies on the immune system following methods were used:

1. To determine the weights of spleen and thymus.

Among those that were slain by the method of the cranio-cervical dislocation animals took the thymus and spleen was determined by their weight on torsionic scales, and then found a weighting factor according to the formula:

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2. Determining the number of antibody productive cells (AFC).

The effect of herbal remedies on the primary humoral immune response was evaluated by counting of AFC in the spleen according to the method of N. K. Eine and A. A. Nordin (1963) monitor lizard (EB). Mice were immunized by intraperitoneal introduction 5 108EB by 20.0 mass mouse in 0.5 ml of isotonic sodium chloride. On day 5, animals were killed, spleens homogenized in 10 ml of Hanks solution (NGOs "plant"), the suspension was passed through a filter. Made calculations of splenocytes in the camera Goriaev and counted their number in the whole spleen. Mix to fill the cells were prepared by adding 0.6 ml of Hanks solution, 0.1 ml of 10% suspension of DL in physiological solution, 0.1 ml of the suspension of splenocytes and 0.2 ml of a compliment Guinea pigs (NGOs the plant), diluted in the ratio 1: 5 isotonic sodium chloride. Then the cells are incubated in a thermostat at 37oC for 1 h After incubation was calculated the number of zones of hemolysis in each cell that corresponds to the number of the KLA. Calculates the number of the KLA throughout the spleen and 106the splenocytes.

3. Cellular immune response was studied on the model of hypercapitalist delayed-type (GST) to sheep erythrocytes by P. H. Lagrange et al (1974). Mice were senzibilizirani subcutaneous injection of 2 to 107EB in 0.2 ml of physiological sodium chloride solution. After 4 days, the animals were injected under the skin of the right foot 108EB in a volume of 0.02 ml left about cervical dislocation, cut off the legs at the level of the ankle joint and weighed on a torsion elastic scales. Then found % weight gain experienced legs compared to control.

4. The functional activity of macrophages and neutrophils was evaluated in NBT-test. Mice received blood and suspension peritoneal macrophages by adding heparin at a final concentration of 50 U/ml

For the production of spontaneous NBT-test) was mixed with equal volumes of blood or suspension of peritoneal macrophages and 0.1% solution narasinga of tetrazole in buffered saline solution, incubated at 37oC for 30 minutes and Then cooked preparations, which were stained with methyl green. For the production of induced NBT-test took 0.05 ml of blood or suspension of peritoneal cells in 0.05 ml suspension of Candida albicans and 0.1 ml narasinga of tetrazole. The following stages consistent with the methodology of spontaneous NBT-test.

Each drug was calculated 100 polymorphonuclear leukocytes or macrophages and determined how many of them are the inclusion of deformation dark blue color that corresponded to the percentage of activated phagocytes.

5. The phagocytic activity of macrophages and neutrophils and is certain mice heparinised buffered saline solution. As a source of neutrophils used heparinised blood. The suspension of cells was mixed with a suspension of killed Candida albicans in physiological solution at a ratio of 10 microbial cells in 1 leukocyte and incubated at 37oC for 30 minutes and Then from a mixture smears were prepared, fixed and stained by Romanovsky-Institute. Calculated % phagocytophila of cells and the intensity of phagocytosis (phagocytic index) - number of Candide, absorbed one phagocyto (Paster E. U. and others, 1989).

The experimental results were processed statistically using the criterion Student. Experiments conducted on 286 outbred mice of both sexes. Herbal drug introduced enterline at doses of 3 and 300 mg/kg, in one series was used dose of 30 mg/kg of the Reference drug is served prodigiosin at a dose of 30 μg/mouse (PL. 3-8).

Studies have shown that the introduction of the drug after immunization EB as once and for 4 days caused a decrease in weight and the weight ratio of the spleen. Observed reduction of the appropriate indicators for the thymus (table.3) with the exception of the weighting factor for thymic at 4 times the introduction of 300 mg/kg of the drug. The introduction of the drug to immunization not have had the mode of administration (table.4). This allows us to conclude that the connection does not show lymphotoxin action, and change other parameters, such as perfusion of the organ. On humoral immune response emitted by the drug has a suppressor effect in the introduction to inductive phase (PL.4). At the same time, the drug does not change the number of AFC in the spleen during the course of the introduction, both before and after the introduction of the antigen that is associated with effects on T and not on B-lymphocytes, when administered within 4 days after immunization (inductive and productive phase) depression of the immune response is not detected.

To clarify the mechanism of action of the drug on the primary humoral immune response to the drug was administered enterline at a dose of 300 mg/kg one hour after antigen. As hypoimmune used a dose of sheep red blood cells 2107by 20.0 mouse narkomanii 2108, hyperimmune 2109(Sibirjak S. V. et al, 1991). In the performed series of experiments, significant inhibition of the immune response in the control was observed only when using hypoimmune sensitizing dose. Hyperimmune dose caused only a tendency to decrease number of AFC in the spleen. The drug, providing a suppressor effect in terms of normoalbuminuria on the number of KLA as the .5).

Studies have shown, four the introduction of the drug after sensitization was significantly suppressed the reaction GST (PL.6). Minimum dose caused a tendency to suppression GST in a single application. Four-time administration of the drug at a dose of 300 mg/kg reduced the ability of polymorphonuclear leukocyte to adapt under the influence of a stimulant (Candida albicans), which is manifested in the suppression of the stimulated NBT-test (table.7). A single injection of phytomedicine no effect on NBT-test, as a compensatory response does not have time to develop.

The peroral dose of 300 mg/kg for 4 days prior to the determination of phagocytosis had no effect on neutrophil and macrophage phagocytosis (Candida albicans) (table.8).

Thus, the obtained data testify to the immunomodulatory activity of herbal remedies.

The anabolic action of herbal drug was studied on 18 Mature rats male Wistar rats, which were divided into 3 groups of 6 each. group 1 animals received inside the drug at a dose of 200 mg/kg, group 2 - methyluracil in the same dose within 30 days, and group 3 was intact. Then, animals were weighed and scored by decapitation, collected blood from the corresponding effect of the drug was tried to increase the weight and mass m. levator ani to change the content of protein in the serum and in the muscle (table.9). Protein was determined by Lowry (Kolb C. R., Kamyshnikov C. C., 1976).

Thus, phytomedicine statistically significantly increases muscle mass and protein level in the blood, and the body weight of animals.

It is established that the proposed phytomedicine has no embryotoxicity.

1. Phyto-nutrient steps for oral administration containing plant extract and fillers, characterized in that as the plant extract contains a condensed extract of alfalfa in the following ratio of components, wt.%:

The condensed extract of alfalfa - of 50.0 to 90.0

Fillers - 9,6 - 53,0

2. Phyto under item 1, characterized in that the filler contains calcium gluconate, sucrose, calcium stearate and talc in the following ratio of components, wt.%:

The condensed extract of alfalfa - of 50.0 to 90.0

Calcium gluconate - 4,0 - 25,0

Sucrose - 5,0 - 25,0

Calcium stearate - 0,2 - 1,0

Talc - 0,4 - 2,0 e

 

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