Antiallergic agent

FIELD: medicine; pharmacology.

SUBSTANCE: antiallergic agent is produced by toluene extraction of the birch bark and contains the active components such as betulin, lupeol and O-caffeate taken in particular quantities.

EFFECT: agent is efficient against allergy and doesn't reveal side effects.

11 tbl, 9 ex

 

The technical field

The invention relates to the field of medicine and more specifically to the search for new means of control actions.

The level of technology

On the socio-economic impact, impact on health and quality of life of allergic diseases are one of the leading places in the structure of human pathology and represent global medico-social problem. According to the literature, present the problem of allergies - the problem of national security, 25-40% of the Russian population have different allergic diseases over the last 30 years in economically developed countries, the incidence of allergic complications doubling every ten years. The socio-economic costs of allergic diseases in Russia is $ 2.6 billion.$ per year.

There are quite a number of widely known antihistamine drugs used in modern medicine, however, the problem of the search for new effective agents against various diseases of allergic origin continues to be relevant. Especially important is the task of identifying new protivoallergennyh natural means, in particular of vegetable origin, with minimal side effects when used.

In folk medicine for a number of plants has protivovesa is positive and anti-allergic properties [http://eastbook.by.ru/minaeva/g14/d24.htm], including figures and birch, indicating the possibility of their use for the preparation of antiallergenic from the leaves of birch and birch SAP.

Anti-inflammatory properties of birch bark possesses due to its constituent Pyh the triterpenoids: betulin and lupeol, uvaol, Betulinol and oleanolic acids and other minor components. Allergenic activity of products derived from this plant, is associated with the content of betulin (see patent RU 2254032), and other triterpenoids - oleanolic acid.

Anti-allergic activity of triterpenoids (as blockers H1histamine receptors) is disclosed by Japanese authors (JP 10-287582), which were examined in this direction bark extract, consisting of the triterpenoids, with the prevalence among them of betulin. However, in the aforementioned work does not reveal the composition of the studied extracts and indicates a different mechanism of action.

The invention

The objective of the present invention to provide a control means on the basis of the composition of triterpenoids with a different mechanism of action than the known allergen antihistamines, and studied composition in the above patents.

Another objective of the present invention is the obtaining of such control means, who, without being certain indicators such the most effective drugs as suprastin and claritin (loratadine), does not give their inherent side effects and more active than the company.

Solved the above problems by the present invention by providing a composition triterpenes certain structure, namely (wt.%):

betulin65-71
lupeol12-16
3-O - caveat betulin5-15
related compounds to100%

The composition is obtained by extraction of the bark - top birch bark method patent RU No. 2192879 belonging to the applicant of this application.

Example 1.

Extraction of the powdered bark was carried out with toluene for about 1.5 hours with constant stirring and the temperature of 90-110°C. After completion of the extraction process was carried out by filtering through the mass of the fabric, at a temperature of 40-50°C.

Of the meal was partially distilled toluene, and the remaining mass is cooled within 6-10 hours to a temperature 15-5°C, after which they were re-fabric filtration and washing of the mass on the filter with toluene. Then was carried out by vacuum distillation of toluene and received the final product in the form of yellowish crystals. who received the extract was subjected to high performance liquid chromatography (HPLC) under the following conditions: column Separon S 6× C18(3,3·150 mm), eluent: acetonitrile/methanol/ water+phosphoric acid 40/40/20+0.6 ml per 100 ml of the mixture, the flow rate of eluent 0.5 ml/min

Detection was carried out using UV detector at 210 nm. The composition of the extract according to the results of HPLC (%):

betulin - 65

lupeol - 12

3-O-caveat betulin - 15

related compounds - the rest is up to 100%

Example 2.

Got an extract of example 1 and subjected to additional cleaning with ethyl alcohol, then washing with water and drying. To receive crystals from cream to white. (If additional treatment increased the content of betulin and lupeol and decreased content Kopiatu betulin, gave yellow crystals). According to the results of HPLC extract had the following composition (%):

betulin - 71

lupeol - 16

3-O-caveat betulin - 5

related compounds - the rest is up to 100%

Examples of research antiallergic activity

The assessment of specific allergenic action of the composition according to the invention were conducted according to the guidelines approved by the Pharmacological Committee of the Russian Federation "guidelines for the assessment of the allergenic properties of pharmacological substances" and "guidelines for the study of immunotropic activity of pharmacological substances" (2000). In this work, we used male Guinea pigs albinos the nursery of the Russian Academy of medical Sciences (RAMS) "Central" weighing 250-300 g and mice male line of the IAS weighing 18-20 g from nursery RAMS "Pillar", after two weeks of quarantine in vivarium research Institute of pharmacology of RAMS. Animals were kept under standard conditions with free access to feed and water. As the main feed were used piketirovany pelleted feed. Comparators were selected two effective drugs with different mechanisms of allergen pharmacological action: claritin (loratadine) and suprastin (chloropyramine).

Research methods

The total reaction anaphylaxis in Guinea pigs-albino

Given the current idea that allergic sensitization, stimulation of production of IgE antibodies is called selective development of Th2-helper cells, as well as experimental evidence of allergenic properties of ovalbumin (13)was selected as the model of sensitization intact Guinea pigs of 0.6% solution of protein chicken eggs (CJD), the major allergenic component of which is ovalbumin.

To obtain anaphylaxis reactions of animals from the control and experimental groups were immunized orally with a 0.6% solution of protein chicken eggs (CJD) in the dose of 1 ml per 250 g of body weight for three days (dissolved in physiological solution CJD Guinea pigs administered orally through a soft plastic tube (6).

In the first series of experiments, 12 days after the achala immunization CJD experimental animals for three days in a/b was introduced 1st experimental group of Guinea pigs studied composition at a dose of 50 mg/kg, The 2nd experimental group of animals was injected drug comparison, suprastin, at a dose of 50 mg/kg, control animals were injected with 1% starch gel. On the following day the 1st experimental group of Guinea pigs was injected studied composition at a dose of 50 mg/kg, 2nd experimental group - suprastin, at a dose of 50 mg/kg, control animals were injected with the same volume of solvent. After 2 hours, the animals of all groups intracardiac introduced CJD in the dose of 1 mg per 300 g body weight, and then recorded the development of anaphylactic reactions.

In the second series of experiments, 12 days after the beginning of immunization CJD experimental animals for three days in a/b was introduced 1st experimental group of Guinea pigs studied composition at a dose of 50 mg/kg, 2nd experimental group of animals was injected betulin dose of 50 mg/kg, control animals were injected with 1% starch gel. On the following day the 1st experimental group of Guinea pigs was injected into/b studied composition at a dose of 50 mg/kg, 2nd experimental group - betulin dose of 50 mg/kg, control animals were injected with the same volume of solvent. After 2 hours, the animals of all groups intracardiac introduced CJD in the dose of 1 mg per 300 g body weight, and then recorded the development of anaphylactic reactions.

Additionally, it was studied the effect of a/W introduction the comparison drug, claritin, dose of 30 mg/kg, anaphylactic reaction in Guinea pigs, immunity avannah 6% solution of CJD.

Calculation of anaphylactic index Weigle was carried out according to the following formula:

,

where N is the number of Guinea pigs, whose death;

N1the number of Guinea pigs that have developed severe shock;

N2the number of Guinea pigs who had developed a mild shock.

N3the number of Guinea pigs who had developed a weak shock.

N4- Guinea pigs who have not come of shock.

When the death of all animals in the group index Weigle is 4(++++). In severe shock 3(+++), moderate shock - 2(++), weak shock - 1(+), in the absence of anaphylactoid reactions in Guinea pigs - index 0 (3).

The reaction of inflammation on the concanavalin

The reaction of inflammation on the line And (psevdoallergicakie reaction) based on the ability of the Con And non-specific, i.e. without participation of the reagents with allergens on the membranes of target cells, as the result of direct action on receptors on the membranes of mast cells and basophilic leukocytes to release inflammatory mediators (histamine, serotonin, leukotrienes, and others).

The experiments were conducted on mice male line of the IAS weighing 18-20 g were Used two routes of administration of test substances intraperitoneal and oral administration, the introduction was carried out in 1% starch gel. Intraperitoneal injection of the investigated composition was administered in doses of 5 mg/kg, 50 mg/K and 100 mg/kg, betulin was administered in doses of 10 mg/kg and 100 mg/kg, Comparators, suprastin and claritin, were administered in doses of 5 mg/kg and 50 mg/kg by oral route of administration studied composition was administered in doses of 5 mg/kg, 50 mg/kg and 100 mg/kg, betulin was administered in doses of 50 mg/kg and 100 mg/kg, suprastin and claritin - in doses of 5 mg/kg and 50 mg/kg Animals of the control group in a similar way have introduced the appropriate amount of starch gel. Given the pharmacokinetic data from study pharmacologically effective compounds, Con A (20 μl of a solution at a concentration of 5 mg/ml) was administered to mice in the experimental and control groups after 2 hours subplantar (in the ball of the rear foot), in the contralateral limb, the same volume of saline. An hour later, the mice were scored, we determined the mass of legs and calculated the index of the reaction of inflammation (Andp) by the formula:

,

where Ropis the mass of the foot, the pad which was introduced by Kohn And;

Ptois the mass of the foot, the tip of which was injected with saline.

Research results antiallergenic action of the composition according to the invention showed the presence of a control biological activity, as evidenced by the following examples.

The influence of the composition of the invention and betulin on exudative edema of the foot rats on carragenan in sravnenie is with dexamethasone

In experiments were used outbred white male rats weighing 170-200 g Animals three experimental groups for 60 min before the injection of carrageenan in b/W was injected composition at a dose of 50 mg/kg, betulin dose of 50 mg/kg) or dexamethasone at a dose of 1.4 mg/kg, which when recalculated for rats E.J.Freirech et al. (5) corresponds to a daily dose for a human is 15 mg of the Control animals was similarly introduced a 1% solution of starch. Then the animal experimental and control groups in the right ball of the rear foot was injected with 0.1 ml of 1% solution of carrageenan (Sigma), in the left control the paw - 0.1 ml of solvent (saline). During the next 4 hours the animals of the control and experimental groups every hour micrometer measured swelling of the feet of the hind limbs. In addition, after 4 hours of experience, immediately after slaughter of animals both hind paws cut off the ledge of the bones below the junction of the small and the tibia and above the heel joint. The severity of acute exudative inflammation was assessed at the end of the experience on the mass difference test (Pop) and control (Pto) feet in the conventional method.

Study of the effect of the composition according to the invention for anaphylaxis isolated bodies on the model of anaphylactic contraction of smooth muscle of the isolated preparation of the ileum (the ileum) Guinea pigs

Guinea pigs albinos weighing 250-300 g were senzibilizirani three Perekrest lisovanim by ovalbumin (Sigma). Ovalbumin was administered three times a day in the form of a mixture consisting of 0.25 ml of physiological solution containing 5 mg of ovalbumin and 0.25 ml of complete adjuvant's adjuvant (Difco). The first injection was made subcutaneously in the region of the hind limbs; two subsequent intramuscularly in the thigh. The experiments were placed in 4 weeks after the last sensitizing injection.

For the preparation of the segments of the intestine of animals was scored by cervical dislocation, dissected abdominal cavity and was isolated ileum, which was placed in Krebs solution. Then cut out the segments of the ileum length of about 2 cm, washed the contents of the intestine with Krebs solution and was allocated the top layer of longitudinal muscles. Thus obtained sample of the longitudinal muscle of the ileum were fixed at one end to the holder with platinum electrodes, transferred into the working chamber with Krebs solution, with the temperature of 37°and the constant passing through a solution of Carbogen (95-96% O2, 5-4% CO2). The second end of the sample was fixed on the sensitive element of mechanotronic. After a 20-minute stabilization period in the periodic update of the Krebs solution in the chamber to confirm the viability of the sample to the electrodes was applied rectangular pulses with a frequency of 0.1 Hz, a duration of 1 MS and a voltage of 0 C. Then recorded the reduction of the sample of the longitudinal muscle of the ileum of the Guinea pig induced by a solution of histamine (10-7M) (7) in the absence or presence of the compounds under study. Reduction of the drug were recorded using mechanotronic converted electrical signals were recorded on a tape recorder. After registration, the effect of the incubation solution in the cell was replaced with a flowing system. The measurements were performed at least three segments of the ileum of each animal.

In the first series of experiments, investigational drugs, composition, suprastin and loratadine, was introduced into the solution after obtaining stable reactions to histamine for 5-15 minutes before the next injection of the agonist. The composition was dissolved in 96,6% ethyl alcohol was added so that the final concentration of the drug was 2×10-3mg/ml In the interval between adding the analyte change Krebs solution in the chamber was made every 5 minutes

In the second series of experiments, the composition is incubated with ileum within one hour and within one day (in Krebs solution, the concentration of the studied drug was 4×10-2mg/ml) at room temperature, to the control samples of ileum was added the appropriate amount of solvent. Next conducted ODA is a division of contractile activity of longitudinal muscle of the ileum, caused by histamine release.

The determination of the activity of NF-kB

Cell line U937 were cultured in cell DMEM medium containing content of bovine serum 10% and before the experiment was planted in 24-hole Board 200 cells/well. Using TransFast transfection reagent (Promega, USA) according to the Protocol of the manufacturer of the cells were transferrable reporter gene luciferase under the promoter containing NF-kB binding elements (pNF-KB-LUC; Stratagene, USA), and the vector containing the gene β-galactosidase (pSV-β-galactosidase control vector; Promega, USA; to assess the efficiency of transfection).

48 hours after transfection vector pSV-β-galactosidase cells were washed in PBS and fixed in 2% formaldehyde/0.2% glutaraldehyde the solution prepared in fosfato-buffered saline (pH 7.4)for 5 min at 4°C. For the analysis of activity β-galactosidase cells were washed in PBS and incubated with the substrate for β-galactosidase o-nitrophenyl-β-D-galactopyranoside at 37°to the development of yellow color. The degree of coloration, which determines the efficiency of transfection, cheated on microplankton reader (PerkinElmer Wallac 1420 VICTOR2 Multilabel Counter) at a wavelength of 420 nm. After transfection plasmid pNF-kB-LUC cells were added to the inventive composition at a final concentration of 50 μg/ml, 10 μg/ml, 5 μg/ml and 1 μg/ml and incubated with him when C and 5% CO2. After 1 hour, the cells were added TNFα(100 ng/ml) and incubated for another 3 hours under the same conditions. After this period, cells were washed in PBS, was destroyed lytic buffer (Promega, USA) and centrifuged at 12000g (1 min, 4°). The supernatant was collected in a sterile 0.5-ml microtubes and frozen to -70°C. To evaluate the activity of luciferase supernatant (5 μg dissolved protein) were made in 96-well polystyreny tablet (PerkinElmer Wallac) with 10 μl/well of reporter lyse buffer. Luminometer (PerkinElmer Wallac 1420 VICTOR2 Multilabel Counter) was programmed for inclusion in each well 50 μl Luciferase Assay Reagent (Promega) and measuring the light intensity after 2 seconds after application. All samples were placed in three Parallels. The activity of nuclear factor kV expressed in terms of activity β-galactosidase (which determines the efficiency of transfection) to the number of conventional light units obtained from the evaluation of the activity of luciferase. The purpose of the correct assessment of the results of the experiment were placed two controls: cells transfected with plasmid pFC-MEKK with constitutive active luciferase gene (positive control), and cells transfected with the plasmid pCIS-CK (Staratagene, USA) with the absence of NF-kB-binding elements in the gene for luciferase (negative control).

Statistical processing of data

Preparation of primary data and the calculations were performed in the environment of a statistical software package (PSP) STATSTICA (version 6.0) for WINDOWS. The results are presented as arithmetic mean and its average error. The significance of differences on the t-criterion of student.

Example 3.

The data presented in table 1, show that/b introduction of the composition at a dose of 50 mg/kg 2 times suppressed the intensity of systemic reactions anaphylaxis in animals of experimental groups compared with control. Index reaction in control group of animals was 2.1 (4 Guinea pigs identified severe anaphylactic shock, 3 - moderate shock, 3 - weak shock). At a dose of 50 mg/kg in 3 pigs developed a mild shock, 4 weak shock, 3 - anaphylactic reactions have not revealed (index reaction by Weigle was 1.0). When in b/W the introduction of the comparison drug, in this case, suprastin (the most effective drug in the treatment of severe forms of arginine)at a dose of 50 mg/kg in 2 animals identified moderate anaphylactic shock, 2 pigs developed a weak shock, the 6 - anaphylactic reactions have not revealed (index reaction by Weigle was 0.6).

Example 4.

In the second experiment also found that in/b introduction the composition of the invention at a dose of 50 mg/kg (table 2) 2 times suppressed the intensity of systemic reactions anaphylaxis in Guinea pigs. The index of the reaction at a dose of 50 mg/kg was 0.8 (1-x Guinea pigs identified severe anaphylactic shock, 2 - mind the i.i.d. shock, the 1st is a weak shock at 6 anaphylactic reaction was absent). When in b/W the introduction of betulin in a dose of 50 mg/kg in 2 pigs developed severe shock, 2 - weak shock, 5 - anaphylactic reactions have not revealed (index reaction by Weigle amounted to 0.9). In the control group of animals 1-a Guinea pig died, 2 - severe shock, in 2 animals identified moderate anaphylactic shock, 5 pigs developed a weak shock (index reaction by Weigle (1.9).

When studying the influence/b introduction claritin dose of 30 mg/kg on the intensity of the anaphylactic shock in Guinea pigs immunized with 6% of CJD, found that in the experimental group of 7 animals 2 Guinea pigs were killed, 2 developed severe shock, in 3 pigs has defined a moderate shock, the index of the reaction was 2.9. In the control group, 7 out of 11 animals died, in 2 animals developed severe shock, 2 - moderate shock index reactions to 3.5 (table 3).

Thus, in all the above experiments, the composition according to the invention is 2 times reduces the severity of systemic reactions common anaphylaxis compared with control groups. Inhibition of anaphylactic shock with the introduction of the composition are more pronounced than the introduction of betulin and the comparison drug claritin, are widely used in various allergic diseases.

Example 5.

From the data, as the x in table 4, it follows that a single intraperitoneal injection of the composition of the invention at doses of 5 mg/kg, 50 mg/kg and 100 mg/kg in mice CBA caused significant suppression of the reaction of pseudoallergy at Stake And 45.3%56,0% and 71.7%, respectively. In a single b/W the introduction of betulin in doses of 5 mg/kg and 50 mg/kg was observed suppression of the reaction of pseudoallergy at Stake And 50.3% (p<0,01) and 39.0% (p<0,01), respectively. Introduction similarly comparative drugs claritin and suprastin, statistically significantly inhibited the reaction of inflammation on the line A. With the introduction of claritin in doses of 5 mg/kg and 50 mg/kg difference from the control values were reached 79,9% and 90.6%, and with the introduction of suprastin in the same doses and 78.6% 66,0%.

Example 6.

Single oral administration of the composition according to the invention at a dose of 50 mg/kg in mice CBA led to a pronounced suppression of the reaction of inflammation on the line And by 72.7% (table 5). A single injection per os betulin in doses of 50 mg/kg and 100 mg/kg and suprastin in doses of 5 mg/kg and 50 mg/kg did not cause suppression of the reaction of inflammation on the line A. the comparator Drug, claritin, administered orally in doses of 5 mg/kg and 50 mg/kg caused a dose-dependent suppression of the reaction of inflammation on the line And by 45.5% to 83.2%, respectively.

Thus, these allow you to indicate the presence in the composition of the invention allergen protivovospalitel the preliminary steps for intraperitoneal and oral administration, betulin and suprastin when administered orally did not possess anti-inflammatory action.

Example 7.

A single intraperitoneal injection of the composition of the invention at a dose of 50 mg/kg white outbred rats-males resulted in a significant suppression of exudative edema caused by carrageenan in the course of the experiment (table 6, see drawing). Rats of the control group of the local inflammatory response is maximally increased after 3 hours after injection of carrageenan, by this time in animals that were administered the composition according to the invention, was a significant suppression of exudative edema by 25.9%. Intraperitoneal administration of a drug comparison of dexamethasone reduced the edema by 31.5% compared with the control group after 3 hours after injection of carrageenan. With the introduction of betulin significant decrease in severity of pericardial edema by 12.0% was observed only after 2 hours after injection of carrageenan. These findings indicate the presence of a composition more pronounced compared with betulin anti-inflammatory action comparable with the action of one of the most effective anti-inflammatory drugs - dexamethasone.

Example 8.

Not detected significant changes in contractile activity of the ileum of Guinea pigs during exposure HDMI is tion according to the invention in various concentrations with samples of the longitudinal muscle of the ileum for 5 min, 1 hour and 1 day, indicating the absence of direct effect of the composition on the H1 receptors (table 8). Blockers H1-receptors, suprastin and loratadine used in various concentrations, caused a marked suppression of the spasmogenic action of histamine at the terminal ileum of Guinea pigs.

Example 9.

Table 9 shows the baseline data obtained during the experiment.

From the table it is seen that the composition according to the invention inhibits both spontaneous and induced activity of nuclear factor kB. A significant reduction of the activity of NF-kB was observed when added to the cells 5 μg/ml or more, with doses of 5 and 10 µg/ml have similar biological activity and reduce the activity of NF-kB in 2-2 .5 times, while 50 μg/ml bring it almost to zero. The translation of the tested doses of the claimed composition on betulin, shows that 50 μg/ml of the composition is 80 μm betulin, i.e. dose not achievable in vivo. More acceptable is the dose of 5 μg/ml, that is 8 μm betulin. It is 2 times suppresses basal activity of nuclear factor kB and 2.5 times TNFa-induced. The obtained results indicate that the high activity of the claimed composition in respect of nuclear factor kB.

The presented data allow to draw a conclusion about the effectiveness of the composition according to the present invention as the means of control actions, comparable with the action of drugs such as suprastin, claritin and dexamethasone, and thus about the possibility of using it as a control tool for the production of hypoallergenic action. It is important to note that the anti-allergic activity of the composition in the experiments was higher than that of its main ingredient - betulin, which indicates the joint action of the ingredients of the composition.

The mechanism of action of the claimed anti-allergic tools

The basis of the pathogenesis of allergic diseases, developing 1 type hypersensitivity, is the activation of Th2 helper cells and production of cytokines IL-4, IL-5 and IL-13, with subsequent synthesis of IgE antibodies with high affinity for fat cells and basophils. The antigen interacts with a fixed mast cells IgE antibodies, which leads to cell activation and secretion of mediators of allergic (histamine, serotonin and others). However, according to the classification P.G.H.Gell and P.R.A.Coombs there are 4 types of hypersensitivity, which are based on other mechanisms. The mechanism of action of classical antihistamines allergen preparations based primarily on the blockade of the binding of histamine to its receptor H1expressed on the cell surface. Antihistamine medicines have the higher the affinity with H 1the receptor, which leads to the preferential binding of the receptor with the drug, and not with histamine (1,4).

It was established experimentally that the mechanism of action of anti-allergic composition according to the invention differs from classical antihistamines and specified in a previously published patents, and the authors Express certain assumptions about the actual mechanism of action.

When studying the effect of the composition according to the invention for anaphylaxis isolated bodies on the model of anaphylactic contraction of smooth muscle of the ileum (the ileum) Guinea pigs are not detected significant changes in contractile activity induced by histamine, ileum of Guinea pigs at different time of exposure of the composition at different concentrations with samples of the longitudinal muscle of the ileum, indicating the absence of direct effect of the claimed composition on H1receptors (15).

In accordance with various mechanisms of pathogenesis of allergic diseases one of the main classes in the treatment of allergies in addition to antihistamine medications are corticosteroids. Steroids have proven to be highly effective against allergic and other types of inflammation, inhibiting both early and late manifestations. For anti-inflammatory effects of corticosteroids, the most important is x the ability to inhibit the synthesis of proinflammatory cytokines, prostaglandins, cellular phospholipases and adhesion molecules (1,4). In particular, activation of phospholipase A2cleavage of arachidonic acid from the phospholipids of cell membranes by cyclooxygenase leads to the synthesis of prostaglandins and lipoxygenase - to the synthesis of leukotriene family, part of which was formerly known as slow-acting substance of anaphylaxis (1). Currently searching steroide-like compounds with the ability to inhibit the activity of phospholipase A2thereby reducing the availability of substrate for the development of various forms of inflammation and with less toxicity compared with classical corticosteroids(4, 7, 9, 14). A number of authors include triterpenoids lupanovo number to the so-called phytoecdysteroids (phytoecdysteroids) due to the fact that the mechanism of action and chemical structure of triterpenoids and steroids are close enough (15, 16). Corticosteroid-allergenic drugs for chronic use lower resistance to infections, disrupt metabolism, particularly the metabolism of carbohydrates, fats and proteins, inhibit the activity of the hypothalamic-pituitary-adrenal axis, causing liver damage, neurological disorders, and have a number of other side effects. Steroide-like compounds of plant origin have t the fir side effects (16).

Anti-inflammatory activity of the compositions according to the invention primarily due to the fact that according to the literature betulin as corticosteroids is an inhibitor of phospholipase A2(7). In addition, according to the literature betulin and betulin acid have affinity for receptors glucorticosteroids, have anti-inflammatory activity comparable with dexamethasone and anti-inflammatory activity of Betulinol acid on the model of edema on carragenan more pronounced than that of betulin (17). In the presented experiments, the composition according to the invention had a more pronounced compared with betulin anti-inflammatory action comparable to the homologue hydrocortisone - dexamethasone.

As part presents the composition of the invention has a large number of triterpenoids of Kopitov, which have antioxidant, protivoopuholevymi, antimalarial and other properties(8, 10, 16, 19). In particular, the antioxidant properties of the extracts of the bark of birch trees is explained by the presence in their composition of coffeecow betulin, Cleanaway and Betulinol acids (5). It is proved that the group kofola (caffeoyl group) enhances the biological activity (18), in this case-allergenic.

According to modern concepts, selective stimulation of Th1helper cells that produce interferon γ (FN-γ ), inhibits the production of Th2cells secreting interleukin IL 4, which stimulates the synthesis of IgE antibodies (13). It is possible that suppression of systemic reactions anaphylaxis are also detected in the compositions according to the invention is the ability to selectively induce γ-interferon and thus hinder the development of Th2helper cells and the synthesis of IgE antibodies (18). Currently it is one of the promising directions in immunotherapy of allergic diseases.

It is possible to suggest another mechanism of action based on the results of a study to identify the composition according to the present invention is the ability to suppress both spontaneous and induced activity of nuclear factor kB. Adding to the cell line U937 composition in the amount of 5,10 mg/ml decreased the activity of NF-kB in 2-2,5 times. High activity in respect of nuclear factor kB may explain the pronounced anti-inflammatory, and some forms of antiallergenic effect.

In the next phase of the research was to study the clinical effectiveness and safety of use of the compositions according to the invention. For this purpose use Superantigens 50 and Superantigens 25" in bronchial asthma (BA). Both bad at its core containing composition according to the present invention. BAD Super is nicox 50 contains 50 mg of the composition and fructose, "Superantigens 25" - 25 mg of the composition and fructose.

Methods and scope of research

Clinical testing BAD "Superantigens 25 and Superantigens 50" conducted on the basis of 5 therapeutic (allergic) Department of the MUSES GKB №57 of Moscow.

Inclusion criteria of patients in the study were:

age from 18 to 60 years,

confirmation of the diagnosis of "asthma" clinical and instrumental methods of examination, consent to treatment and examination,

the absence of concomitant diseases that have a significant impact on the course of the underlying pathology and requiring additional assignments (diabetes, tuberculosis, malignant tumors).

The study included 39 patients with bronchial asthma.

To assess the clinical effectiveness of drugs in treatment of patients developed original scheme diaries of medical observation and questionnaires for patients in whom the clinical manifestations of the disease was assessed in points.

The use of drugs agreed upon by all patients received the informed consent of each patient.

In accordance with the Protocol, a survey of patients:

Laboratory:

1. Standard clinical analysis of a blood (the General).

2. Determination of immunoglobulin E.

3. Cytologies the e study of induced sputum and nasal secretions (in bronchial asthma and allergic rhinitis).

Instrumental:

- Spirography, including the test with the use of bronchodilators.

- Perfluorotri.

- Determination of nitric oxide in exhaled air.

The studies were conducted before and after treatment.

Brief characteristics of patients with bronchial asthma

The study included 39 patients with bronchial asthma of varying severity in the acute stage.

When admitted to hospital, all patients were observed typical clinic of bronchial asthma (BA) with violation of the General conditions (to the state average severity), increased breathing rate (from 21 to 30 min), change of percussion sound (boxed tint, boxed) and the emergence of auscultatory changes (breathing weakened, hard; wheezing with various quantitative characteristics).

Laboratory examination of patients conducted in accordance with the General standards and the study Protocol. In General, the analysis of blood in 10 cases (5 in each group) showed increased level of leukocytes to 12.9-13,4·106, mainly in patients received corticosteroids, often (4 cases), moderate eosinophilia - up to 8%.

The amount of immunoglobulin E when intake was increased in 25 of the 39 patients; the level of increase was different from 187 UNITS to more than 1000 IU(threshold method).

Patients divided into two subgroups depending on the intended treatment investigational BAD: "Superantigens 50" (21 patients) or Superantigens 25" (18 patients). Then each subgroup divided taking into account clinical evidence of systemic glucocorticosteroids (SGX).

In one case the patient on the 6th day of taking the "Superantigens 50" appeared urticarnae rash, extinct after withdrawal of the drug. The exact relationship with the drug difficult to ascertain, because in those days the patient ate different food, including allergenic nature. This patient in group observations were not included.

Thus, observed the following groups:

I gr. (10 patients) received "Superantigens 50" background SGX,

II gr. (10 patients) received "Superantigens 50" without SGX,

III gr. (9 patients) received "Superantigens 25" background SGX,

IV gr. (9 patients) received "Superantigens 25" without SGX.

Patients of groups I and II from the first day of hospital stay were appointed "Superantigens 50": 1 capsule 2 times a day, 15-20 minutes before meals; daily dose of 100 mg treatment ranged from 15 to 20 days depending on duration of stay in the hospital.

Ill III and IV groups from the first day of hospital stay were appointed "Superantigens 25": 1 capsule 2 times a day, 15-20 minutes before meals; daily dose is 50 mg. Course l the treatment ranged from 15 to 20 days.

As a basic treatment patients received, depending on the severity of the disease, broncholytic and inhaled corticosteroids (IGX) (beclamethasone dipropionate, fluticasone propionate in doses recommended GINA 2002). On the background of exacerbation and hospitalization in the hospital 19 patients received SGX.

In group II, 8 patients out of 10 received only a "Superantigens 50", two patients also had ingalirovti broncholytic; IGCs was not appointed. Thus, this group can be considered as monotherapy group.

In group IV only Superantigens 25" received 1 patient, the combination with bronholitikami - three, the combination with IGCs and bronholitikami - five patients.

The control group (16 patients, 8 with the inclusion of SGX without them) received only basic therapy.

To evaluate the clinical effect was used diaries medical supervision and the original questionnaire for the sick.

In the diary observations physician daily was evaluated available leading symptoms of asthma in points:

I Shared state:

- Satisfactory - 3 points

- Moderate - 2 points

- Heavy - 1 point

II respiratory rate:

- 20 - 0 points

- 21-25 - 1 point

- 26-30 - 2 points

- 31 and more - 3 points

III Percussion:

- pulmonary - 1 point

with boxed shade - 2 points

- boxed - 3 points

p> IV Breath:

- vesicular - 1 point

- weakened - 2 points

- hard - 3 points

V Rales:

no - 0 points

- single whistling during forced exhalation - 1 point

- a few whistling on the exhale - 2 points

- multiple whistling, humming - 3 points

Patients also assessed their health, noting its in the questionnaires at 1, 5, 10 days of treatment and at discharge (15-20 days) (in points):

I. General condition

1 point: very poor

2 points: bad

3 points: satisfactory

4 points: good

5 points: excellent

II. The frequency of asthma attacks (once a day, week, month)

III. Cough

0 points: no

1 point: some coughing

2 points: daily moderate

3 points: rare paroxysmal

4 points: paroxysmal daily

5 points: constant, persistent cough that interfere with sleep

IV. Shortness of breath

0 points: no

1 point: rarely under load

2 points: rarely alone

3 points: constant

V. wheezing when breathing

0 points: no

1 point: only a limited period of time (morning, evening), with exercise

2 points: often, several episodes during the day

3 points: constant

VI. The feeling of stuffiness in the chest

0 points: no

1 point: only a limited period of time (morning, evening), when the physical load

2 points: often, several episodes during the day

3 points: constant

In the assessment of clinical effectiveness study SUPPLEMENTS were taken into consideration objective manifestations of BA, the results of the survey of patients, reflecting the health and functional abilities of the patients in taking medications, and laboratory and instrumental data.

The results of the STUDY EFFICACY BAS IN COMPLEX THERAPY of BRONCHIAL ASTHMA

"Superantigens 50"

"Superantigens 50" were appointed in complex treatment of 20 patients with bronchial asthma. In the study group included 16 women and 4 men aged from 18 to 60 years; the average age was 45.3±2, 87.

Diagnosed forms of bronchial asthma: atopic - 8, endogenous - 6, mixed - in 6 patients. Current: slight persisting - in 5, moderate in 12, heavy in 3 patients (Table 10).

When referring all patients had typical clinical picture of BA corresponding forms and severity.

In group I - patients with initial severe disabilities requiring assignment SGX positive dynamics of clinical manifestations of ad, due, mainly, hormonotherapy, came quickly. Average time normalization of the General condition (from moderate to satisfactory) was 3.0±0,31 day, the disappearance of od is the loud - 3,1±0,90 day. Percussion was pulmonary, on average, 6,3±1.47 days, auscultatory picture normalized later: vesicular breath were heard through 3-6-10 days (average: 7,6±1.69 days), dry rales disappeared through 5-17 days (average - by 9,7±1.76 days).

In the control group of patients treated with SGX (8 people), these figures were: normalization of the General state - 3,6±0.45 days, the disappearance of dyspnea - 4,0±0.97 day, normalization of percussion sound - 7,4±1.51 day, auscultatory picture 8,2±1.51 day for breathing, 10,2±1.84 days for the disappearance of wheezing. Thus, the indicators in the main group was normalized a bit faster, which can be noted as a trend (without statistical significance).

In group II the original condition of the patients was slightly better: a state of moderate severity detected in 3 of 10 patients, normalization of it happened for 1.4±0.28 days (control - 2.4±0.30 day; the significance of differences with a p<0,05). Respiratory rate in 6 cases was initially normal, and the rest is normalized, on average, for 6 days (control - 6, 2 days). Percussion sound with boxed shade was observed in 8 patients, normalization occurred within 6-11 days, and 4 patients boxed shade survived until discharge from the hospital (in the control - similar). Vesicular respiration is their source was found in 3 patients, others in the treatment process, it became vesicular - on average, 13.4±0,18 day (control - 14.1±0,31 day). Dry rales were auscultatory all patients at admission; the timing of their disappearance ranged from 7-8 to 15 days, for an average of 10.3±0.81 days; control and 11.6±0.92 days.

"Superantigens 25"

"Superantigens 25" were appointed in complex treatment of 18 patients with bronchial asthma. In the study group was composed of 11 women and 7 men aged from 18 to 60 years; mean age was 44,3±3,22 year.

Diagnosed forms of bronchial asthma: atopic - 5, endogenous - 4, mixed - in 9 patients. Current: slight persisting - in 6, moderate - 8, heavy - 4 patients (table 11).

When referring all patients had typical clinical picture of BA corresponding forms and severity.

In group III patients receiving SGX and Superantigens 25 on the background of moderate and severe AD, the General condition of the source was assessed as moderate, normalization has come through 2-8 days, average - by 3,7±0,31 day. At the same time normalized breathing rate. Normalization percussion painting took 8,6±2,02 day, the normalization of breathing - 10,7±1.84 days, the disappearance of wheezing - 10,7±1,62 day. In the control group experienced a similar phenomenon (see above).

In patients of group IV (similar to II gr.) overall the condition, respiratory rate and percussion original picture were little changed, their normalization occurred in 5-6 days. However, 2 patients boxed shade percussion, as in group II, survived until discharge, as well as dry rales.

The average duration of clinical manifestations were: violation of the General state - 1,9±0.29 days, changes in breathing rate to 5.0±1.91 days, changes of percussion sound is 6.5±1.67 days of breathing patterns to 11.2±2,14 days, dry rales in the lungs - 8,9±1,na.

Analysis of clinical results of monotherapy with drugs. In group II patients actually received monotherapy "Superantigens 50" (only in 2 cases out of 10 supplemented with bronholitikam faradila). Individual assessment of well-being and objective status of patients before and after treatment have made it possible to observe positive results in half of the patients (5 cases), moderate positive dynamics - 3, no objective dynamics - in 2 patients. In group IV ("Superantigens 25") about the true monotherapy can speak only in one case; positive dynamics of clinical manifestations of ad. The remaining patients were used broncholytic (3; all with positive symptoms), or a combination of broncholytics with ihx (5, 2 of them - with the positive changes of all parameters, 3 - positive Dina is IR subjective parameters).

Changes in the health and condition of patients in the treatment process was monitored daily as the attending physician and by the patients.

Analysis of objective and subjective indicators in patients with BA who received "Superantigens 50 and Superantigens 25" on a background basis-treatment with the inclusion SGX (I and group III)showed that the main therapeutic effect, of course, had SGX. Some differences in the speed of normalization of the General condition marked by prolonged intake of dietary supplements: 10-20 days of treatment self-assessment by patients of their condition was slightly higher than in the control group (p<0,05).

In the group of patients treated without the use of SGX, there are some differences in the occurrence of positive changes in objective and subjective parameters. When alone "Superantigens 50" (group II, 10 patients) on the 20th day of treatment (before discharge from hospital) awarded the best auscultatory picture (normalization of breathing, the disappearance of wheezing (p<0,05). Patients on the 20th day of treatment noted in the questionnaires better feeling compared with the control, 10 and 20 days of treatment, the disappearance of difficulty in breathing (p<0,05).

In group IV to consider the use of "Superantigens 25" monotherapy is not possible, since the majority of patients received a bronchodilator, and half of the patients also, and beclomethasone. However, it should be noted that certain significant differences in clinical parameters from control in these patients referred to the same parameters: a greater number of patients already on day 5 auscultative vesicular breathing had returned to normal health (judging by the scores at the 5th, 10th and 20th days of treatment). Similarly to group II shortness of breath, wheezing and congestion of the chest disappeared in the earlier period (p<0,05; p<0,01).

Thus, the results of the clinical analysis showed that the use in the treatment of patients with BA preparations BAS "Superantigens 50 and Superantigens 25" helps somewhat earlier, in comparison with control, normalization of well-being, and in applying it for 10-20 days and some indicators of breath (auscultatory and subjective data).

Portability "Superantigens 50 and Superantigens 25". Drugs Superantigens 50 and Superantigens 25", in General, was well tolerated by the patients. Noted above case of allergic reaction (urticaria), is not significantly correlated with the use of "Superantigens 50".

The results of clinical and laboratory-functional observations allow us to recommend BAD "Superantigens 50 and Superantigens 25" for use in complex therapy of Bo is lnyh bronchial asthma. According to clinical observation, both tools contributed to an earlier normalization of well-being, as well as some indicators of breath (auscultatory and subjective data). Analysis of laboratory data revealed no negative effects of medication BAS on parameters to be monitored, in some cases there was a positive dynamics of indicators such as eosinophilia, serum total immunoglobulin E level of nitric oxide in exhaled air and indicators of external respiration function (these laboratory values had a tendency to normalization, is not statistically confirmed).

Table 1
DoseThe number of animals in the groupIndex of response WEIGLE
Control102,0
Composition:

100 mg/kg

1000 mg/kg
10

10
1,1

1,3
Table 2
DoseThe number of animals in the groupIndex of response WEIGLE
To the Troll 102,1
Composition 50 mg/kg101,0
Suprastin: 50 mg/kg100,6
Table 3
DoseThe number of animals in the groupIndex of response WEIGLE
Control101,9
Composition:

50 mg/kg
100,8
Betulin: 50 mg/kg90,9
Table 4
DoseThe number of animals in the groupIndex of response WEIGLE
Control113,5
Claritin 30 mg/kg72,9

Table 5
The dose of the pharmacologically effective compounds, theThe number of animals in the groupIndex reaction% of control
Control1015,9± 1,7100
Composition:
5 mg/kg108,7±1,9 (p<0,05)54,7
50 mg/kg107,0±1,1 (p<0,01)44,0
100 mg/kg104,5±1,4 (p<0,01)28,3
Betulin:
5 mg/kg107,9±1,2 (p<0,01)49,7
50 mg/kg109,7±1,3 (p<0,01)61,0
Claritin:
5 mg/kg

50 mg/kg
10

10
3,2±1,1 (p<0,01)

1,5±0,5 (p<0,01)
20,1

9,4
Suprastin:
5 mg/kg103,4±1,1 (p<0,01)21,4
50 mg/kg105,4±1,7 (p<0,01)34,0

Table 6
DoseThe number of animals in the groupIndex reaction
To ntrol 1013,8±1,6
Composition: 5 mg/kg1012,4±2,2
Control1014,3±2,0
Composition:
50 mg/kg

100 mg/kg
10

10
3,9±1,3 (p<0,01)

10,6±2,1
Betulin:
50 mg/kg1014,6±2,5
100 mg/kg1014,0±2,5
Claritin:
5 mg/kg

50 mg/kg
10

10
7,8±2,2 (p<0,05)

2,4±0,8 (p<0,01)
Suprastin:
5 mg/kg

50 mg/kg
10

10
16,4±1,9

13,7±3,3

Table 7
Time measurement of swelling (in hours)Swelling of the foot ×10-1mm
Control (n=8)Composition 50 mg/kg,/b (n=8)Betulin 50 mg/kg,/b (n=8)Dexamethasone 1.4 mg/kg,/b (n=8)
033,3±1,434,8± 1,632,0±0,929,8±0,7
143,5±1,937,3±1,6*38,3±0,8*32,0±0,9**
249,0±4,036,8±1,6*41,8±2,836,0±0,8**
353,7±5,939,8±1,4*44,8±2,336,8±1,1*
452,3±5,737,0±1,6*41,5±2,030,7±1,1**
Note: the significance of differences from control: * p<0,05; ** - p<0,01; n - number of animals in the group

td align="center"> Loratadine 0.01 mg/ml
Table 8
GroupThe exposure time of connectionsContractile activity of longitudinal muscle portion of the ileum (%)
Control100
Track5 min89,1±3,8
Suprastin 1×10-10mg/ml5 min46,5±4,1
Suprastin 2×10-3mg/ml5 min26,5±1,5
Suprastin 0.2 mg/ml5 minFull suppression
15 min28,4±3,2
Control (alcohol)1 hour100
BAS1 hour90,1±17,1
Control (alcohol)24 hours100
BAS24 hours108,3±11,2

Table 9

SampleLuciferase (intensity of illumination)β-galactosidase (OD 420 nm)Luciferase/β-galactosidaseLuciferase/βGal.
Rooms holes123123123(M±m; n=3)
Cells without stimulation4705605070,070,0620,0586714903787478166±1201
Composition (1 µg/ml)2223566150.0360.0390.0646175912396148304±1766
Composition (5 µg/ml)153 1081830.0350.0330.054360327236603764±523
Composition (10 µg/ml)1531221570.050.0370.043060330539353433±428
Composition (50 µg/ml)3.73.41.70.0020.0020.0011843172216521739±92
Control (TNFα)731.590912540.0590.0690.05812398131742162115731±4858
BAS (1 μg/ml) +TNFα7586003900.0570.050.051330012002794911084±2650
Composition (5 μg/ml) +TNFα1876064130.0480.0730.0643890829664586214±2102
Composition (10 μg/ml) +TNFα1107733149 0.1010.0417964725636446288±2200
Composition (50 μg/ml) +TNFα3530.0020.0040.0031327115511541212±94
Positive control182413687760.0750.0650.03524320210462216322510±1580
Negative control0.91.11.40.0490.0480.04419233124±6

Table 10
Group I (n=10); Superantigens 50+basis-treatment with the inclusion SGXGroup II (n=10) Superantigens 50+basis-treatment without the use of SGX
Form BA:
Atopic44
Endogenous24
Mixed42
The severity of ASTHMA:#x0200A;
Easy persisting for14
Moderate66
Heavy3-

Table 11
Group III (n=9); Superantigens 25+basis-treatment with the inclusion SGXGroup IV (n=9) Superantigens 25+basis-treatment without the use of SGX
Form BA:
Atopic23
Endogenous31
Mixed45
The severity of ASTHMA:
Easy persisting for-6
Moderate53
Heavy4-

Literature:

1. Gushchin I.S. - Allergic inflammation and its pharmacological control. - M, Farmersbranch, 1998, s-180.

2. Deryabin p. g, Spout N.N., E.I. Isaev, Kondrashin N.G., Inozemtsev A.N., Presnov GA, Topoleva T.V. - interferon-inducing properties of birch bark extract, soda is containing betulin, and its effect on the infection caused by hepatitis C virus in tissue culture cells and in the body of the infected mice - Problems of Virology RAMS, 2005, No. 5, s-16.

3. ADO VA, Goryachkin L.A. - the Suppression of allergic reactions of low-molecular compounds - Minsk, 1971, p.40.

4. Manual immunopharmacology edited by Dale M.M., Formen D.K., M, Medicine, 1998, 246-259.

5. A pointer NM, Tsymbal, I.N., Petrenko NI, etc - Intermolecular interactions dipeptide derivatives of triterpenoids and natural quinones with α-tocopherol. - Biomed. chemistry, 2003, v.49, issue 1, p.26.

6. Hlopushina YEAR, Krinsky A.V., Kovalenko, L.P. and other - Impact of sikarin on the pharmacokinetics of antipyrine in the intact and sensitized Guinea pigs - bul. the experts. Biol., 1991, No. 7, p.67-69.

7. Bernard P, Scior T, Didier, Hibert M, Berthon J.Y. - Ethnopharmacology and bioinformatic combination leads for discovery: application to phospholipase A(2) inhibitors - Phyto-chemistry, 2001, Vol.58, p.865-874.

8. Cheng J., Bai, Y.J., Zhao Y.Y., Wang, C., Cheng M. - Studies on the phenylpropanoids from Eucoinmia ulmoides - Zhongguo Zhong Yao Za Zhi, 2002, Jan; 27(1), p.38-40.

9. Chumkaew P., Kato, S., Chantrapromma, K. - A new triterpenoid ester from the fruits of Bruguiera parviflora - Chem Pharm Bull (Tokyo), 2005, Jan; 53(1), p.95-96.

10. Da Cunha P.M., D. Duma, Assreuy J. Buzzi F.C. et al. - Caffeic acid derivatives: in vitro and in vivo anti-inflammatory properties - Free It. Res., 2004, Nov; 38(11), p.1241-1253.

11. Faquim-Mauro E.L., Macedo M.S. - Induction of IL-4-dependent, anaphylactic-type and of IL-4-Independent, non-anaphylactic-type IgG1antibodies is modulated by adjuvants - Int. Immunology, 2000, Vol.12, No. 12, p.1733-1740.

12. Francis G., Krem Z, Makkar H.P.S., Becker K.-The biological action of in saponins in animal systems: a reviev - Brit. J. Nutrition, 2002, Vol.88, p.587-605.

13. Kimber I., N.I. Kerkvliet, Taylor S.L. et al. - Toxicology of protein allergenicity: prediction and characterization - Toxicol. Sci-1999, Vol.48, P.157-162.

14. Klein R. - Phytoecdysteroids - J. of the American Herbalists Guild (JAHG), Fall/Winter 2004, p.18-28.

15. Merlos m, Giral M., D. Balsa - Rupatadine, a New Potent, Orally Active Dual Antagonist of Histamine and Platelet-Activating Factor - J. Pharmacol. Exp. Ther., 1997, Vol.280, Issue 1, p.114-121.

16. Nuntanakorn P., Jiang C., Einbond L.S. et al. - Polyphenolic Constituents of Actaea racemosa - J Nat. Prod., 2006, 24, 69(3), R-318.

17. Patocka J. - Biologically active pentacyclic triterpenes and their current medicine signification - J. of Applied Biomedicine, 2003, No. 12, ISSN 1214-0287, p.7-12.

18. Wang, K.W., Sun C.R., X.D. Wu, Y.J. Pan - Novel Bioactive Dammarane Caffeoyl Esters from Celastrus rosthornianus - Planta Med., 2006, Mar., 72(4), p.370-372.

19. Watkins R.W., Tumey D.B. - A report for the forestry commission and the government-industry forum for non-food crops - a Review of current knowledge on the economic potential of chemical products from the main commercial tree species in the UK, CSL, July, 2003, p.17-19.

The remedy for allergies, obtained by toluene extraction of birch bark, containing as active components betulin, lupeol, 3-O-caveat betulin and related substances the following amount, wt.%:

betulin65-71
lupeol12-16
3-O-caveat betulin5-15
related substancesthe rest is up to 100%



 

Same patents:

Indanol derivatives // 2323937

FIELD: chemistry.

SUBSTANCE: invention relates to novel compounds of general formula (I): (where R1 and R2 may be identical or different, and each is a 1,3-substituted aryl with substituents from group α; R3 stands for any of the following groups: -CO-R4, -CO-O-R4, -CO-NH-R4, -CO-CH2-N(Ra)Rb, -(CH2)m-CO-R5, -(CH2)m-R5, -CO-NH-CO-N(Ra)Rb, -CO-NH-SO2-N(Ra)Rb, -CO-NH-CO-(CH2)m-N(Ra)Rb, or -CO-NH2; R4 stands for a lower alkyl, cycloalkyl, cycloalkyl substituted with 1-3 substituent from group α, lower alkenyl, lower alkynyl, halogen-substituted lower alkyl, hydroxyl-substituted lower alkyl, lower alkoxyalkyl, lower aliphatic acyloxyalkyl or lower alkoxycarbonylalkyl; R5 stands for hydroxyl, -OR4 or -N(Ra)Rb; Rа and Rb may be identical or different, each of them stands for hydrogen, hydroxyl, lower alkoxy group, hydroxyl-substituted lower alkoxyl, hydroxyl-substituted lower alkoxyalkyl, lower alkoxy lower alkoxyalkyl, cyano lower alkyl, cyano lower alkoxyalkyl, carboxy lower alkyl, carboxy lower alkoxyalkyl, aliphatic lower alkoxycarbonyl lower alkoxyalkyl, carbamoyl lower alkyl group, carbamoyl lower alkoxyalkyl, lower aliphatic acylamino lower alkyl, lower aliphatic acylamino lower alkoxyalkyl, lower alkylsulphonylamino lower alkyl, lower alkylsulphanylamino lower alkoxyalkyl, (N-hydroxy-N-methylcarbamoyl) lower alkyl, (N-hydroxy-N-methylcarbamoyl) lower alkoxyalkyl, (N-lower alkoxy-N-methylcarbamoyl) lower alkyl, (N-lower alkoxy-14-methylcarbamoyl) lower alkoxyalkyl or R4, or both, including associated nitrogen, stand for nitrogen-containing heterocyclic group or nitrogen-containing 1-3 substituted heterocyclic group with substituents from group α; m is an integer from 1 to 6; А stands for carbonyl; В stands for straight bond; D stands for oxygen atom; Е stands for С14 alkylene; n is an integer from 1 to 3; and α group is a group of substituents, which consist of halogen atoms, lower alkyls, hydroxy lower alkyls, halogen lower alkyls, carboxy lower alkyls, lower alkoxyls, hydroxy lower alkoxyls, hydroxy lower alkoxyalkyls, lower alkoxycarbonyls, carboxyls, hydroxyls, lower aliphatic acyls, lower aliphatic acylamines, (N-hydroxy-N-methylcarbamoyl) lower alkyls, (N-lower alkoxy-N-methylcarbamoyl) lower alkyls, hydroxy lower aliphatic acylamines, amines, carbamoyls and cyano groups), or pharmacologically suitable salt thereof. Invention also relates to pharmaceutical composition and method for disease prevention and treatment.

EFFECT: preparation of novel biologically active compounds.

18 cl, 117 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel compounds of formula I or pharmaceutically suitable salt or solvate thereof, where dashed line stands for additional bond, а is a number from 0 to 2, b is a number from 0 to 2, n is 2, p is 2, r is 1, М1 stands for nitrogen, М2 stands for С(R3), X stands for either a bond or alkylene group with number of carbon atoms from 1 to 6, Y stands for -С(О)- group, Z stands for a bond, or alkylene group with number of carbon atoms from 1 to 6, or alkenylene group with number of carbon atoms from 1 to 6, or -С(O)-, -CH(CN)-, -SO2- or СН2С(O)NR4- group, R1 stands for groups, R2 stands for six-membered heteroaryl ring with one or two heteroatoms chosen independently of each other from either nitrogen atom or N-O group, other atoms of the cycle being carbon, five-membered heteroaryl ring with one, two, three or four heteroatoms chosen independently of each other from nitrogen, oxygen or sulphur, other atoms of the cycle being carbon, R32 stands for substituded quinoline group, R32 stands for substituted aryl group, heterocycloalkyl group, cycloalkyl group with number of carbon atoms from 3 to 6, alkyl group with number of carbon atoms from 1 to 6, group, where the said six-membered heteroaryl ring or the said five-membered heteroaryl ring may be R6-substituted, R12 independently of others is chosen from an alkyl group with number of carbon atoms from 1 to 6, hydroxyl group or fluorine atom, provided in case R12 stands for hydroxyl or fluorine the rest of R12 cannot be bonded to a nitrogen-bonded carbon atom, or two R12 substituents form an alkyl bridge with number of carbon atoms from 1 to 2, which bonds two non-adjaicent carbon atoms of the ring, R13 independently of the others is chosen from an alkyl group with number of carbon atoms from 1 to 6, hydroxyl group, alcoxy group with number of carbon atoms from 1 to 6, or fluorine atom, provided in case R13 stands for hydroxyl or fluorine the rest of R13 cannot be bonded to a nitrogen-bonded carbon atom, or two R13 substituents form an alkyl bridge with number of carbon atoms from 1 to 2, which bonds two non-adjacent carbon atoms of the ring. See description for meaning of the other structural elements. Invention relates also to pharmaceutical compositions, as well as to application of compounds of formula I.

EFFECT: preparation of novel biologically active substances and pharmaceutical compositions.

20 cl, 659 ex

FIELD: organic chemistry, medicine, neurology, pharmacy.

SUBSTANCE: invention relates to novel hydrogenated pyrrolo[4,3-b]indoles of the general formula (1): , their racemates, optical isomers, geometric isomers, pharmaceutically acceptable salts and/or hydrates that can be used, for example, in treatment and prophylaxis of different neurodegenerative diseases, such as Alzheimer's syndrome. In the general formula (1): a dotted line with accompanying unbroken line represents ordinary or double bond; R1 and R2 represent independently of one another substitutes of amino group chosen from hydrogen atom, possibly substituted (C1-C6)-alkyl substituted possibly with aryl, possibly substituted phenyl, possibly substituted carbonylamino or thiocarbonylamino group, substituted acyl, possibly substituted aryl sulfonate wherein substituted in indicated R1 and R2 are chosen from (C1-C6)-alkyl, halogen atoms, nitro, carboxy, alkoxy group, aryl; R1n represents one or some similar or different substituted of cyclic system chosen from hydrogen atom, alkyl, aryl, cyano group, halogen atom, 5-6-membered nitrogen-containing heteroaryl. Also, invention relates to methods for synthesis of these compounds, pharmaceutical compositions and their using, and to using compounds in libraries with their using.

EFFECT: valuable medicinal properties of compounds and pharmaceutical compositions, improved methods of synthesis.

20 cl, 2 tbl, 12 ex

FIELD: pharmaceutics.

SUBSTANCE: the present innovation deals with developing compositions for preventing or treating pollinosis, allergic rhinitis, atopic dermatitis, asthma or urticaria. Composition for preventing or treating pollinosis, allergic rhinitis, atopic dermatitis, asthma or urticaria containing two types of medicinal raw material out of Cucurbita moschata and Carthamus tinctorius seeds and, at least, one medicinal raw material chosen out of Plantago asiatica, Lonicera japonica, Coix lachrymal-jobi var. ma-yuen. Composition for preventing or treating pollinosis which should be applied for introduction for a pollinosis-suffering mammal, annually before pollinosis season that contains two types of medicinal raw material out of Cucurbita moschata and Carthamus tinctorius seeds and, at least, one medicinal raw material chosen out of Plantago asiatica, Lonicera japonica, Coix lachrymal-jobi var. ma-yuen as an efficient component. Method for obtaining a composition for preventing or treating pollinosis, allergic rhinitis, atopic dermatitis, asthma or urticaria containing two types of medicinal raw material out of Cucurbita moschata and Carthamus tinctorius seeds and, at least, one medicinal raw material chosen out of Plantago asiatica, Lonicera japonica, Coix lachrymal-jobi var. ma-yuen deals with the following stages: plant raw material should be mixed, extracted; the extract should be purified and concentrated and, if necessary, one should add the fillers followed by granulation. Method for obtaining a composition for preventing or treating pollinosis, allergic rhinitis, atopic dermatitis, asthma or urticaria containing two types of medicinal raw material out of Cucurbita moschata and Carthamus tinctorius seeds and, at least, one medicinal raw material chosen out of Plantago asiatica, Lonicera japonica, Coix lachrymal-jobi var. ma-yuen deals with mixing plant raw material, if necessary, with the fillers, the mixture obtained should be mixed with a plastifying solvent followed by granulation and drying. Method for preventing or treating pollinosis deals with the fact that a pollinosis-suffering patient should be annually introduced before pollinosis season with a composition that contains two types of medicinal raw material out of Cucurbita moschata and Carthamus tinctorius seeds and, at least, one medicinal raw material chosen out of Plantago asiatica, Lonicera japonica, Coix lachrymal-jobi var. ma-yuen. Dietetic food product or functional food product for preventing or treating pollinosis, allergic rhinitis, atopic dermatitis, asthma or urticaria obtained due to introducing two types of medicinal raw material out of Cucurbita moschata and Carthamus tinctorius seeds and, at least, one medicinal raw material chosen out of Plantago asiatica, Lonicera japonica, Coix lachrymal-jobi var. ma-yuen as an efficient component. Dietetic food product or functional food product for preventing pollinosis that contains two types of medicinal raw material out of Cucurbita moschata and Carthamus tinctorius seeds and, at least, one medicinal raw material chosen out of Plantago asiatica, Lonicera japonica, Coix lachrymal-jobi var. ma-yuen as an efficient component. Dietetic food product for animals or functional food product for animals for preventing or treating pollinosis, allergic rhinitis, atopic dermatitis, asthma or urticaria containing two types of medicinal raw material out of Cucurbita moschata and Carthamus tinctorius seeds and, at least, one medicinal raw material chosen out of Plantago asiatica, Lonicera japonica, Coix lachrymal-jobi var. ma-yuen as an efficient component. The above-mentioned compositions and dietetic products are highly efficient for preventing or treating pollinosis, allergic rhinitis, atopic dermatitis, asthma or urticaria.

EFFECT: higher efficiency of prophylaxis or therapy.

23 cl, 9 dwg, 15 ex, 2 tbl

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention elates to novel derivatives of imidazolidine of the formula (I): , wherein symbols B, E, W, Y, R, R2, R3, R30, e and h have values given in claim 1 of the invention. Compounds of the formula (I) represent pharmaceutically active substances. Compounds of the formula (I) relates to inhibitors of adhesion and migration of leukocytes and/or antagonists of adhesion receptor VLA-4 that relates to group of integrins. Proposed compounds can be used in treatment of diseases that are induced or associated with undesirable degree of leukocyte adhesion and/or migration of leukocytes, or wherein interaction between cells or between cells and matrix are very significant and important and these interactions are based on interaction of VLA-4 receptors and ligands. Except for, invention relates to methods for synthesis of compounds of the formula (I) and pharmaceutical compositions containing compounds of the formula (I).

EFFECT: valuable medicinal properties of compounds and pharmaceutical composition, improved method of synthesis and preparing.

12 cl, 19 ex

FIELD: organic chemistry, chemical technology, medicine, pharmacy.

SUBSTANCE: invention relates to novel hydrogenated azepino[4,3-b]indoles of the general formula (1): , their racemates, optical isomers, geometric isomers and their pharmaceutically acceptable salts and/or hydrates. In the general formula (1) a dotted line with accompanying unbroken line represents a simple or double bond; R1 and R2 represent independently of one another substitutes of amino-group chosen from hydrogen atom, possibly substituted (C1-C8)-alkyl possibly substituted with aryl, 5-6-membered azaheterocyclyl, (C1-C8)-alkoxycarbonyl, possibly substituted phenyl, possibly substituted carbonylamino- or thiocarbonylamino-group, substituted acyl, (C1-C8)-alkylsulfonyl, possibly substituted arylsulfonyl and wherein substitutes in indicated R1 and R2 are chosen independently from (C1-C8)-alkyl, halogen atoms, nitro-, carboxy-, alkoxy-group, aryl; Rin represents one or some similar or different substitutes of cyclic system chosen from hydrogen atom, (C1-C8)-alkyl, (C6-C10)-aryl, halogen atom, 5-6-membered azaheterocyclyl. Also, invention relates to methods for synthesis of these compounds, their using and pharmaceutical composition and libraries of compounds. Synthesized compounds possess neuroprotective, cognitive-stimulating and anti-histaminic properties and can be used in treatment of different neurological disorders, allergic and autoimmune diseases, for example, for memory improvement.

EFFECT: valuable medicinal and biological properties of compounds and pharmaceutical composition, improved method of synthesis.

25 cl, 2 tbl, 12 ex

FIELD: medicine, dermatology, pharmaceutical industry, pharmacy.

SUBSTANCE: invention relates to a composition used in treatment and prophylaxis of allergic diseases and non-allergic inflammatory diseases in mammals and containing winter-hardy kiwi extract. Also, invention relates to a method for preparing the composition containing winter-hardy kiwi extract and involving diluting milled and dried winter-hardy kiwi with water or lower alcohols, heating a diluted winter-hardy kiwi and extraction of product to obtain winter-hardy kiwi extract. Also, invention relates to a method for treatment, relive or prophylaxis of allergic diseases and non-allergic inflammatory diseases in mammals involving administration in a mammal winter-hardy kiwi extract for treatment or prophylaxis of at least one symptom of allergic disease or non-allergic inflammatory disease. Also, invention elates to a method for treatment and prophylaxis of allergic diseases and non-allergic inflammatory diseases in mammals involving prescription or administration in mammal winter-hardy kiwi extract for treatment or prophylaxis of at least one symptom of allergic disease or non-allergic inflammatory diseases in mammal. Also, invention relates to using winter-hardy kiwi extract for treatment, relieve or prophylaxis of allergic diseases or non-allergic inflammatory diseases in mammals. Above described winter-hardy kiwi extract and a composition based on thereof show effectiveness in treatment, relieve or prophylaxis of allergic diseases or non-allergic inflammatory diseases in mammals.

EFFECT: valuable medicinal properties of composition.

58 cl, 5 dwg, 11 tbl, 11 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to novel compounds of the formula (I): wherein X represents nitrogen atom (N) or -CH; R1 represents cycloalkyl, aryl, heterocycle, aralkyl, heterocyclic alkyl or oxy-derivative, or group of the formula: ; R2 represents -NR4R5, -OR4; R3 represents tetrazole, -CN, -CH2OH or -CO-R7; R4 represents hydrogen atom (H), -G1-R8 or group of the formula: or wherein G1 represents -CO, -CH2, -SO2; R5 represents H, (C1-C4)-alkyl; R7 represents hydroxy-, amino-, hydroxylamino- or oxy-derivative or amino-derivative; R8 represents aryl, heterocycle, cycloalkyl, aralkyl. Also, invention describes a pharmaceutical composition designated for treatment of VLA-4-dependent inflammatory diseases based on proposed compounds. Also, invention considers using these compounds for treatment and preparing an agent for treatment of VLA-4-dependent inflammatory diseases. Invention provides synthesis of novel compounds possessing useful biological properties.

EFFECT: improved and valuable medicinal properties of compounds and pharmaceutical composition.

26 cl, 9 tbl, 5 ex

FIELD: medicine, allergology.

SUBSTANCE: invention relates to treatment of allergic diseases, in particle, bronchial asthma. Invention proposes using black rice extract or pelargonidine, or cyanidine-glycoside for development of a therapeutic agent. These agents are used in the developed therapeutically effective doses. Invention provides effective treatment based on inhibition of accumulation of eosinophyles in cells of damaged tissues and organs.

EFFECT: enhanced and valuable medicinal properties of composition.

13 cl, 1 tbl, 4 dwg, 6 ex

FIELD: organic chemistry, pharmaceuticals.

SUBSTANCE: invention relates to new chemical compound of general formula I , or salts, or hydrates thereof. In formula I R1 represents group of formula -G1-R1a (wherein G1 represents single bond, oxygen, sulfur; R1a represents C1-C10-alkyl optionally substituted with halogen or C3-C8-cycloalkyl); R2, R3 and R4 are independently hydrogen or -G20-R20 (wherein G20 represents single bond, oxygen, sulfur, sulfinyl or sulfonyl; R20 represents C1-C6-alkyl optionally substituted with 1-3 halogen atoms or C3-C8-cycloalkyl); R5 and R6 are independently -X5-X6-X7 group (meanings of X5, X6 and X7 are as defined in specification) or R3 and R4 may together form pyrrol ring optionally substituted with C1-C6-alkyl; Ar represents phenyl, 1,3-benzodioxolyl, naphthyl, pyridyl, optionally substituted with 1-3 substituents (as defined in specification) Compounds of formula I has antagonistic activity in relates to receptors of corticotrophin releasing factor (CRF). Also disclosed are compounds, characterized by preferable structures, pharmaceutical compositions, using such compounds, intermediates for production thereof and method for treatment of various diseases mediated by CRF.

EFFECT: new compounds as antagonists of CRF receptors.

33 cl, 1 tbl, 316 ex

FIELD: medicine; veterinary.

SUBSTANCE: homeopathic drug Lyarsin is used as a pharmaceutical agent. 0.7 mL of Lyarsin per head is administered intramuscularly to the animals suffering from hepatosis, at the first trimester of pregnancy, twice in 3 days.

EFFECT: method reduces number of stillborn puppies and increases brood per pupped dam.

3 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: film dosage form sticking to mucous, containing at least on active substance from hemp. The form contains the polymeric matrix, which is used as a reservoir for the active substance and sticking to mucous. The form can have the multilayer structure. It can also contain the non-active ingredients, odorants or aromatisors. The film form is used in treatment of pain in such diseases as carcinomatosis, AIDS, trauma and other disorders.

EFFECT: invention provides better tolerance to administered substances and rapid onset of administered drug, simplifies administration procedures.

19 cl

FIELD: medicine.

SUBSTANCE: therapeutic-cosmetic agent for the external use contains pantohematogen, water, polymeric gel-forming component, as well as dimethyl sulfoxide and water. The components are taken in particular quantitative ratio.

EFFECT: agent shows stable efficiency in treatment of osteoarthrosis and spinal osteochondrosis.

7 cl, 2 dwg

FIELD: medicine.

SUBSTANCE: therapeutic-cosmetic agent for the external use contains pantohematogen, water, polymeric gel-forming component, as well as dimethyl sulfoxide and water. The components are taken in particular quantitative ratio.

EFFECT: agent shows stable efficiency in treatment of osteoarthrosis and spinal osteochondrosis.

7 cl, 2 dwg

FIELD: medicine.

SUBSTANCE: invention relates to the compositions for care of the oral cavity, which contain the components of the Indian Mulberry, known as Morinda citrifolia L., for treatment of single or multiple abnormalities of the oral cavity and dental abnormalities, including the periodontal diseases, such as gingivitis and periodontitis, tooth destruction, bad smell and other disorders and irritations of the oral cavity.

EFFECT: composition provides efficient care of the oral cavity.

36 cl, 20 tbl, 11 ex

FIELD: medicine; cosmetology.

SUBSTANCE: invention relates to the agents for elimination of grey hair. The agent contains the chromogenic agent, which contains the beer yeast, hop and water in the following correlation, in mass%: 5-10 of the beer yeast, 2.5-5 of hop, water - the rest.

EFFECT: these components are harmless and provide efficient restoration of dark color in grey hair.

1 tbl, 3 ex

FIELD: medicine; cosmetology.

SUBSTANCE: invention relates to the agents for elimination of grey hair. The agent contains the chromogenic agent, which contains the beer yeast, hop and water in the following correlation, in mass%: 5-10 of the beer yeast, 2.5-5 of hop, water - the rest.

EFFECT: these components are harmless and provide efficient restoration of dark color in grey hair.

1 tbl, 3 ex

FIELD: medicine; cosmetology.

SUBSTANCE: invention relates to the cosmetic agents, namely cosmetic agent for care and maintenance of the natural hair function; contains the routine cosmetic raw materials, differs in containing the 2-furanon derivative , at least one polymeric compound and at least one compound of protein hydrolysates and their derivatives.

EFFECT: invention efficiently maintains natural hair function.

6 cl, 32 ex, 1 dwg

FIELD: medicine; cosmetology.

SUBSTANCE: invention relates to the cosmetic agents, namely cosmetic agent for care and maintenance of the natural hair function; contains the routine cosmetic raw materials, differs in containing the 2-furanon derivative , at least one polymeric compound and at least one compound of protein hydrolysates and their derivatives.

EFFECT: invention efficiently maintains natural hair function.

6 cl, 32 ex, 1 dwg

FIELD: medicine; cosmetology.

SUBSTANCE: invention relates to the cosmetic agents, namely cosmetic agent for care and maintenance of the natural hair function; contains the routine cosmetic raw materials, differs in containing the 2-furanon derivative , at least one polymeric compound and at least one compound of protein hydrolysates and their derivatives.

EFFECT: invention efficiently maintains natural hair function.

6 cl, 32 ex, 1 dwg

FIELD: medicine, oncology, amino acids.

SUBSTANCE: invention relates, in particular, to the development of an antitumor preparation based on natural substances. Invention relates to an amino acid preparation comprising at least one modified essential amino acid obtained by treatment of amino acid by ultraviolet radiation (UV) at wavelength 250-350 nm for 12-80 h at temperature 15-30oC or with ozone at temperature 15-25oC. The modified amino acid has no toxicity for health cells. Also, invention relates to a method for preparing such preparation. Invention provides the development of an antitumor preparation based on modified amino acids and expanded assortment of antitumor preparations being without cytotoxicity for normal cells.

EFFECT: valuable medicinal antitumor properties of preparation.

8 cl, 4 tbl, 2 dwg, 4 ex

Up!