Immunomodulatory means not apply
(57) Abstract:The invention relates to medicine, and is concerned medicines, which have immunomodulatory effects that are used for non-injection use for the prevention and treatment of diseases caused by impaired immune functions. As specified funds proposed the use of erythropoietin. The drug is more effective in comparison with the known analogues, has no contraindications, and largely has focused specialized action, allowing significantly more likely to avoid side effects. table 2. The invention relates to medicine and relates to medico-biological drugs that have anti-inflammatory and immunomodulatory effects, used for non-injection use for the prevention and treatment of any disease due to impaired immune functions. As any pathology is based on or associated with the changing reactions of the immune system, the drug according to the invention can be applied in all areas of clinical medicine, the most vaccinee application involves the use of a medicinal product in all cases, except for the introduction into the tissue and the body cavity is isolated from the external environment. When this drug is manufactured in all possible dosage forms: liquid (e.g., medicine, drops, lotions, rinses, bath, injection, solutions, etc ), solid (e.g., powders, tablets, pills, candles, beads, pellets, granules, sticks, capsules, suppositories, etc), soft (e.g., ointments, pastes, gels, creams, liniments, plasters, applications, etc), gaseous (for example, aerosols, gases, vapors, etc).In accordance with the classification of drugs by drug group drug according to the invention applies to the following pharmacotherapeutic group:
13.5 - Stimulators of hematopoiesis (excluding iron preparations).2 - Tools for the treatment of skin diseases.5 - preparations for treatment of diseases of the organs of the respiratory system.6 - preparations for treatment of diseases of the sense organs.8 - Tools immunomodulatory, immunoglobulins, vaccines and phages.The ability to stimulate the immune response of the organism have derived nucleic acids, as well as a number of biogenic drugs. Sposobnaya for their wide application in complex therapy of infectious diseases and inflammatory diseases, at low current regeneration processes and a number of other diseases.Especially important has become in recent years the study of the immunological properties of endogenous compounds produced by the body itself. These compounds mobilize the immune power of the body to combat pathological processes.One of the most important endogenous immunomodulators are interferons.Known Interferon human leukocyte (Reference Vidal, M.: OVPEE-Attraversare, 2000, C. B-268). Composition: active substance group of endogenous proteins produced by leukocytes of donor human blood under the influence of viruses. Pharmacological action: antiviral, immunomodulatory, antiproliferative. Used for prevention and treatment of influenza, SARS, viral diseases of the eye in the form of an aqueous solution. For treatment of influenza and ARVI interferon instilled into the nasal passages, it can also be used in inhalations.Known human leukocyte Interferon in the candlelight. The active substance group of endogenous proteins produced by leukocytes of donor human blood under the influence of viruses. Pharmacological action: antiviral, immunome immunomodulatory drugs is their heavy portability and have many side effects, in particular: suppression of blood - Interferon alpha N1 (Reference Vidal, M.: OVPEE-Attraversare, 2000, S. E.-178). Not being anti-inflammatory, immunomodulating agents only stimulate the production of proinflammatory cytokines. The drugs Interferon ineffective in Allergy, autoimmunity, as protivovospolitelnyh funds.From the funds immunomodulators not the application should allocate drugs created on the basis of extracts of chamomile (Reference Vidal, M.: OVPEE-Attraversare, 2000, C. B-123, B-205): Kamalogam (108 mg chamomile extract, 1 g of ointment) or Kamillosan (1% ointment dry extract of chamomile flowers), or simply chamomile flowers. As is well known (B. N. Leskov, Chiradeep A. N., Gorlina N. K. The Novozhenov Century, "Clinical immunology for clinicians M., 1997, S. 128), extracts of chamomile have immunomodulatory activity. The disadvantage of the application of these products is their low immunomodulating activity, the difficulty of standardization provided effect in connection with plant origin raw materials. Finally, the effect of chamomile is not specific and are aimed at different parts of the immune system that do not accurately affect specific reaction sys., C. B-290). The active ingredient in this drug - glucosaminilmuramildipeptide representing the main structural unit of the cell wall of bacteria. The drug is used for immunomodulate and has dogowosajy effect. In small doses it stimulates Pro-inflammatory reactions of the immune system (activation of phagocytosis, T-and b-lymphocytes, production of proinflammatory cytokines, such as tumor necrosis factor). In large doses it is used for the treatment of cutaneous autoimmune diseases by inhibition of the synthesis of proinflammatory cytokines. Disadvantages of drug: contraindicated in pregnancy, there are cases of drug intolerance; high probability of allergic reactions in connection with the microbial origin of the product; the ability hyperthermic reactions. As is well known (B. N. Leskov, Chiradeep A. N. , Gorlina N. K. The Novozhenov Century, "Clinical immunology for clinicians", M, 1997, S. 128), the effect of preparations of bacterial origin occurs through Macapagal system, when a result of the activation of antigen presenting macrophage cells produce factors that stimulate immune response. The inhibition of the immune response under the influence of large doses of Licopid due razvitiya develops the condition, similar to the "beyond the break", when under the influence of high doses of antigen is pathological response inhibition. Despite the potential clinical effect, this effect cannot be called physiological and even upon receipt of a positive clinical effect may develop complications related to the imbalance of immunological functions.The closest drug the same destination to the claimed invention, the set of essential characteristics is known immunomodulatory drug Prednisolone (M. D. Mashkovsky. Medicinal product. In two volumes.T. 2.-Ed. 13-e, new. -Kharkov: Torsing, 1997, page 33). Prednisolone is a glucocorticosteroid hormone with pronounced anti-inflammatory and immunomodulatory effects when used in small doses.Indications: used for rheumatism, non-infectious arthritis, bronchial asthma, acute lymphatic and maloletnog leukemia, infectious mononucleosis, neurodermatitis, eczema, and other indications for the use of glucocorticosteroids. Used reinjection in the form of ointments, creams, tablets.Assign Prednisolone orally in the form of t is mikroboy etiology (eczema, itching, dermatiti and so on) using 0.5% prednizolonovuyu ointment, which is applied in a thin layer on the skin. Active substance - Prednisolone (0.005 g).In practice eye (keratitis, conjunctivitis, irity and others) using eye drops containing 0.3% solution of hydrochloride 21-(deoxy-N-methyl-N-piperazinil)-prednisolone. The active substance is a solution of the hydrochloride 21-(deoxy-M-methyl-M-piperazinil)-prednisolone.In small doses (up to 5 mg per day for the average person) prednisolone immunomodulatory effect. Concentration in the media for external use (creams, ointments, drops) is designed to immunosuppression (B. N. Lozova, S. M. Shergin. Structural-functional organization of the immune system. Novosibirsk, Nauka, 1981, 226 S.; B. N. Leskov, Chiradeep A. N., Gorlina N. K. , Novozhenov Century, Clinical immunology for clinicians. M., 1997, 128 S.). However, prednisone is a synthetic analogue of the natural hormone cortisol, so its effect on the immune system has dogowosajy effect.The standard used regimens of prednisolone is used to suppress immunological reactions (including inflammation and allergies), as is a strong Inno-vascular system and hemostasis, from the blood, bone, eye, and including, on the part of the immune system: suppression of protective reactions by lowering resistance to infection, slow healing of wounds. Immunosuppressive drugs can be very effective when applied to overcome tissue incompatibility and treatment of immune diseases, but this property is common to all glucocorticosteroids, prevents the use of prednisolone in acute viral infections, glaucoma, parasitic diseases, erosive and ulcerative processes of the mucous of the upper parts of the gastrointestinal tract (GIT), lymphadenitis, systemic mycosis, etc., i.e. diseases that require stimulation of the immune processes.The disadvantage of prednisolone is also the inadmissibility of prolonged use and abrupt termination, due to habituation to the body. This is because the mechanism of action of prednisone is associated with inhibition of protein synthesis and cell division, and this property is universal with respect to all cells. Hence the braking regeneration, wound healing, increasing the likelihood or stimulation of the infectious process and is of renala effective anti-inflammatory and immunomodulating drugs non-injection use.The problem is solved by the creation of new medicines, with more effective anti-inflammatory and immunomodulatory effects in comparison with the known analogues, no contraindications, and largely has focused specialized action, allowing significantly more likely to avoid side effects.This technical result in the implementation of the invention is achieved in that in the known anti-inflammatory and immunomodulating agent not use containing the active substance and the target additive, as active substances applied erythropoietin.Erythropoietin (EPO) is an important hormone of the body of mammals, including humans, produced by the interstitial cells of the kidney and regulating erythropoiesis in mammals. EPO can be obtained by means of biotechnology - recombinant RAC (recombinant human erythropoietin), by chemical synthesis or can be isolated from human tissue or animal.Recombinant human erythropoietin (RAC) receive, for example, the method of introduction of the cloned gene, erythropoetin the mm wscc/P/D) (patent 2089611). The strain deposited in the special collection transplantable somatic cells of vertebrates of the all-Union collection of cell cultures under number SCC (p) D from 36.04.94.It is established that EPO has anti-inflammatory and immunomodulatory properties (C. A. Kozlov. Clinical immunology in the clinic of internal diseases. Novosibirsk, 1997.- S. 20), affecting the differentiation of fat cells (Irani A. M., Schwartz L. B. Human mast cell heterogeneity // Allergy Proc. - 1994. - Vol. 15, N 6. - R. 303-308), causing an increase in the number of granulocytes and the reduction of T-lymphocytes in the blood (COA T., Narita J., S. Honda et al. Erythropoietin dosage the expansion of c-kit+progenitors for myeloid and erythroid cells, but not for lymphoid cells in the bone marrow and liver // Eur. J. Haematol. - 1999. - Vol. 63, No. 5. - P. 306-312), increasing the sensitivity of tumor cells to the action of PC-cells (Miyajima J. , Imai Y., Nakao M. et al. Higher susceptibility of erythropoietin-producing renal cell carcinomas to lysis by lymphokine-activated killer cells // J. Immunother. Emphasis Tumor Immunol. - 1996. - Vol. 19, N 6. - P. 399-404), by adjusting the ratio of Th1/Th2 dysbalance cells (Matsumoto A., Seki Y. , Kubo M. et al. Suppression of STAT5 functions in liver, mammary glands, and T cells in cytokine-inducible S2-containing protein 1 transgenic mice // Mol. Cell Biol. - 1999. -Vol. 19, N 9. - P. 6396-6407), participating in the regulation of the response of T-lymphocytes to interleukin-2 (Moriggl R., D. J. Topham, S. Teglund et al. Stat5 is required for IL-2-induced cell cycle progression of peripheral T cells // Immunity. - 1999. - Vors on human erythroid colony-forming cells // Exp. Hematol. - 1997. -Vol. 25, N 3. - P. 193-198). Anti-inflammatory effects of EPO due to its influence directly on cell effectors immunopathological (Pro-inflammatory and allergic) reactions, because he is anti-inflammatory cytokine.The advantage of EPO in its high immunomodulatory activity: his current dose of 1000 IU per person per day (about 8-10 IU/kg of body weight) is 0.5 mg (according to the technological regulations 1 mg contains 2000 ME activity), while prednisolone is used at the minimum dose of 5 mg, and in the usual dose is from 30 to 1000 mg.EPO is easily tolerated, by class of toxicity refers to non-toxic substances that do not dose-dependent.Side effects in non-injection use of the new drugs are not available.EPO is standardized in units of activity (ME) and is measured in ME and/or mg Standardization according to the requirements of Pharm. Articles are carried out in vitro by electrophoresis and isoelectrofocusing and in vivo in standard models in animals (linear mouse).EPO has a specific effect. It stimulates the differentiation and proliferation of cells of hemopoiesis in the direction of erythroid Rostock, and in relation cleto is Yaya on other reactions of the immune system (for example, antitumor immunity or anti-infective immunity). This allows accurately to define indications for use of new drugs and to avoid unwanted side effects.To obtain medicines that meets the invention, the active substance - EPO is mixed with the target additive in accordance with accepted pharmaceutical methods of compounding.Target additive - a substance or complex of substances, allowing to obtain the necessary dosage form, do not possess the desired biological activity.The activity and effectiveness of the drug does not depend on the form of targeted supplements, which may have different natural and synthetic forms depending on the dosage form of the drug, as determined solely by the action of the active substance EPO.The drug according to the invention can be applied to children and pregnant women, it does not cause hyperthermia reactions. Allergic reactions are noted only with long-term subcutaneous (parenteral) administration of the medicinal product with the EPO as an active substance. Since EPO has direct selective effect on the individual response systems who we are and not conducive razblokirovka immunological functions.The examples below show the comparative results of drug treatment with the active substance - RAC (recombinant erythropoietin) and drugs with other active ingredients.Example 1.We investigated the efficiency of a drug with the EPO as an active substance in experimental allergic conjunctivitis.Methods: Guinea pigs (n = 68) were sensitized intraperitoneally injection of ovalbumin. 3 weeks after sensitization lower concentration of ovalbumin (10 μg/ml) was administered daily for 3 weeks. 6 weeks to provoke allergic inflammation used 20 mg/ml of ovalbumin.All animals were divided into 3 groups.Group a ( control 1, n = 30), animals are as protector of inflammation were treated with placebo (0,9% solution of sodium chloride, the active substance is sodium chloride).Group b (control 2, n = 22), animals that received drops DEXAPOS (0,1%) eye drops containing 1 mg of active substance - dexamethasone - 21 - sulfobenzoate sodium).Group C experience, n = 16), animals that received 0.1% solution of EPO (pharmaceutical preparations is vospalitelnoe effect was applied for 30 min before the application of the allergen, after 30 minutes, then every hour for 7 hoursAfter 30 min, 8 and 24 h after application of the allergen in the biopsy of conjunctiva to determine the level of inflammatory response.The data obtained are presented in table.1.Found that after 30 min after application of the allergen on the conjunctiva inflammation levels in the control groups and the group, the preprocessed solution with EPO did not differ.After 8 h after application of the allergen is the total number of cells, characterizing the inflammatory process (neutrophils and eosinophils), was significantly different in all groups. In group a this indicator was 213,2 +/- 75.5 cells in field of view. In group - 173.5 metric +/- 28,9, and infiltration overwhelmingly dominated by neutrophils, In the group experience (C) this indicator was 45,8 +/-10,1 cells in field of view.After 24 h after injection of the allergen level of eosinophils was in the control group And 75,7 +/- 15,5, in the control group In at 25.7 +/- 9,9, and group experience - 23,3 +/8.9bn cells/ field of view.The level of neutrophils: in group a - 168,5 +/- 27,8; in group 197,2 +/- 34,7; in the group With - 39,9 +/- 5,5.It should be stated that an objective examination of the animals of both control groups showing signs of purulent conjunctivitis is elite marked anti-inflammatory and antiallergic effect as drugs with EPO as an active substance, and glucocorticoids (dexamethasone), but the effect of glucocorticoids was manifested to a greater extent in respect of eosinophils, but not neutrophils and the total impact on the infiltration was lower. In addition, severe infiltration by neutrophils is one of the signs of suppuration, as confirmed by objective data.Thus, the introduction of the solution with the EPO as an active substance to a large extent can prevent and inhibit inflammation that occurs in experimental allergic conjunctivitis, which confirms its more effective anti-inflammatory effect.Example 2.We examined 14 patients with chronic atopic dermatitis and symptoms of extemalization. All patients were men aged 18 to 49 years. Disease duration ranged from 6 months to 12 years. Patients were examined in the acute phase of the disease. All patients received conventional treatment, including the use of oral antihistaminic preparations, calcium preparations, the proportion of patients with locally advanced 0,5% prednizolonovuyu ointment in patients with severe itch - tranquilizers.All patients were divided into 2 groups: a control group that received EPO (drug active substance EPO).Monitoring the effectiveness of treatment was carried out on the basis of subjective feelings of patients and results of inspection through 14, and 21 days from the start of treatment, then after 3, 6, 12 months.Found that after 14 days of use of the drug EPO in patients group In signs of the disappearance of the inflammatory manifestations. This is manifested in the reduction of local hyperemia, the disappearance of itching, in the case of moist eczema marked decrease in exudation. A positive trend was observed in 7 patients. After the 3 week course of treatment with EPO remission was as follows: up to 3 months in 3 patients; up to 6 months in 1 patient; over the years - in 3 patients.In the control group (group a) after 14 days of treatment, a positive trend was observed in 4 patients after 21 days, another 2 patients. In 1 patient the effect of the treatment were observed. After the 3 weeks of treatment remission was as follows: up to 3 months in 6 patients. In 1 patient, as already mentioned, the effect of the treatment has not been reached.Thus, the external use of medicines with the EPO as an active substance has a significant clinical effect in chronic atopic dermatitis. A comparative study of the influence of the clinical effect of the medicinal product with EPO, that, obviously, is connected with its more specific influence on the pathological reaction of the immune system.Given the applicant's analysis of the prior art, including searching by the patent and scientific and technical information sources, and identify sources that contain information about the equivalents of the claimed invention. Has allowed to establish that the applicant had not discovered similar, characterized by signs, identical to all the essential features of the claimed invention. The definition from the list of identified unique prototype, as the most similar in essential features analogue, has identified a set of essential in relation to perceived technical result of the distinctive features in the claimed medicinal product set forth in the claims.Therefore, the claimed invention meets the condition of "novelty."Anti-inflammatory and immunomodulatory effects created pharmaceutical composition is determined solely new active substance EPO, and other components (target additive) are neutral media from a range of traditionally used in compositions to create the next person, produced by interstitial cells of the kidney and regulating erythropoiesis in mammals. EPO can be obtained by means of biotechnology - recombinant BER (recombinant human erythropoietin), by chemical synthesis or can be isolated from human tissue or animal.To obtain medicines that meets the invention the active substance EPO is mixed with the target additive in accordance with accepted pharmaceutical methods of compounding.Target additive - a substance or complex of substances, allowing to obtain the necessary dosage form, do not possess the desired biological activity.The activity and effectiveness of the drug does not depend on the form of targeted supplements, which may have different natural and synthetic forms depending on the dosage form of the drug (e.g. alcohol, water, oil, gel and other).Below are examples of specific compositions with various forms of the target additives, where the active substance is applied RAC (recombinantly Erythropoietin).Example 3.The pharmaceutical composition representing ointment with EPO as an active substance and fill Beeswax - 4-8
Olive oil - 2-6
Lanolin - 10-20
The method of preparation.The active substance is produced by way of introducing the gene cloned human erythropoietin comprising the plasmid PSVdepoL Mo in cells of transplantable lines of Chinese hamster ovary tk(strain wscc/P/D). Components in the reactor are fed through the dispensers. In the reactor (at t=70oC) melted by sequentially depositing components, starting with the high-melting waxes. The resulting mixture is cooled to t=40oC. Then, the cooled mixture contribute with active stirring, recombinant human erythropoietin. Then make filtering and filling.Example 4.The pharmaceutical composition representing suppositories (rectal, vaginal, urethral sticks, with RAC as active substance and filled with oil-based, containing the following ingredients in the following their ratio, wt.%:
REC - 0,5
Cocoa butter - 2-6
Solid fat basis up to 100
The tool is prepared as follows. Active substance get in the way described above. The components in the machine for the manufacture and packaging of suppositorien. Then make filtering and filling.Example 5.Pharmaceutical composition with RAC as the active substance and the filler gel base containing the following ingredients in the following their ratio, wt.%:
REC - 0,5
Sorbitanoleat - 2-5
Lanolin alcohol - 1-3
High molecular weight fatty alcohols - 4-6
Olive oil - 15-20
Glycerin - 2-4
Water up to 100
The tool is prepared as follows. Active substance get in the way described above. The components in the reactor are fed through the dispensers. In the reactor consistently contribute components and mix. Gelling components are dispersed in water at t=50oC, after swelling and uniform distribution, i.e., the formation of a dispersion. A homogeneous mass is neutralized and cooled with stirring to room temperature. Then with stirring, bring an active substance.Example 6.Pharmaceutical composition, representing a solution for inhalation, with RAC as active substance and filled with water-based, contains the following ingredients in the following their ratio, wt.%:
REC - 0,5
Theophylline - 2-4
oC, the dissolution of sodium chloride, the active substance, sterilizing filtration of the resulting solution, pouring the solution into a bottle and seal.Example 7.The pharmaceutical composition representing eye drops (nose drops), with RAC as active substance and filled with water-based, contains the following ingredients in the following their ratio, wt.%:
REC - 0,01-0,5
Sodium chloride 0,9-1
Water up to 100
The tool is prepared as follows. Active substance get in the way described above. The method of obtaining the claimed dosage forms includes the following sequence of operations: the dissolution of the active substance and sodium chloride in water for injection by heating the solution up to t=50oWith the sterilizing filtration of the resulting solution, filling the solution into a bottle and seal.Thus, the above data confirm that the implementation of the use of the claimed invention the following cumulative conditions:
- farmacevticheskie for use in medicine, namely, for non-injection use for the prevention and treatment of any disease due to impaired immune functions;
- for the claimed medicinal product, as it is described in the independent clause sets out the claims, confirmed the possibility of its implementation using the steps described in the application and known before the priority date tools and techniques;
- a means of embodying the invention in its implementation, is able to achieve the technical result: the creation of a new pharmaceutical composition having a more effective anti-inflammatory and immunomodulatory effects in comparison with the known analogues, no contraindications, and largely has focused specialized action, allowing significantly more likely to avoid side effects. The use of non-injection forms of erythropoietin as an immunomodulator.
FIELD: medicine, otorhinolaryngology.
SUBSTANCE: one should treat deformation in laryngeal and tracheal lumen due to excessive growth of granulation tissue in the sites of their lesions. One should introduce hormonal preparations, moreover, one should apply Diprosan as a hormonal preparation injected once intramucosally at 0.1 ml/sq. cm of granulation tissue, but not more than 0.3 ml.
EFFECT: higher efficiency of therapy.
3 ex, 1 tbl