The method of producing mielopid |
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IPC classes for russian patent The method of producing mielopid (RU 2041716):
 Peptidemia fraction of spleen mammals possessing immunostimulatory activity and its preparation / 2033796
The invention relates to the field of immunology, immunopharmacology and can be used in medicine, veterinary medicine, intensive care, intensive therapy
The method of treatment of purulent and burn wounds / 2016577
The invention relates to medicine, in particular to methods for treating wounds
 Means for stimulation of reparative processes in the liver after surgery on it in terms of the destruction of the body131j / 2005479
The invention relates to medicine, namely to the treatment of liver pathology in terms of the defeat of radionuclides (in particular131I)
 "painkiller "mielopid"" / 2000789
 Method for preventing postsplenectomic syndrome in experiment / 2247567
Embryonic spleen should be sampled, washed in nutritive medium № 199 to be placed into fresh medium № 199 to obtain homogenate in teflon homogenizer followed by centrifuging; then one should isolate the upper, medium and inferior layers, suck off medium layer and the upper part of inferior layer; the cell mixture obtained should be diluted in nutritive medium № 199 to be then introduced by injections into mesentery of small intestine or rectus muscle of abdomen. The present innovation favors the activation of immune system in patients undergone splenectomic operation and in those in case of surgical immunodefficient state due to high functional and regenerating activity of transferred embryonic splenic cells.
 Method for treating the cases of non-operable cardiologic patients / 2250772
Method involves using mononuclear autologic marrow fraction containing 6-9x104 mesenchyma cells per 1 ml or autologic mesenchyma trunk cells. The cells are separated from brain bioptate in the amount of 106 cells/kg of patient body mass. The preparations are intracoronarily introduced in fractions at a rate of 3-5 ml/min into the right coronary artery. The introduction is also carried out in intra-arterial mode in jets or in drops.
Method for treating the cases of allergic diseases / 2250773
Method involves introducing autologic mesenchyma trunk cells as a single intravenous drop dose or in the amount of 1 mln cells per 1 kg of patient body weight.
Method for treatment of suppurative wound / 2257211
Method involves every day covering a wound with an agent comprising of pig spleen homogenate and oxygenated with perfluorane taken in the following amounts: 100 g of pig spleen homogenate and 10 ml of oxygenated perfluorane. Invention promotes to accelerating sanitation of suppurative wounds due to activation of topical immunity and activation of regenerating processes. Invention can be used in treatment of suppurative wounds.
Method for treating noninvasive cancer of urinary bladder / 2257219
The present innovation deals with treating locally spread tumor diseases of urinary bladder due to applying either chemo- and/or radiation therapy. Moreover, one should intravenously once inject by drops autologous mesenchymal stem cells (AMSC) at the quantity of 1 mln cells/kg patient's body weight. Moreover, in case of chemotherapy AMSC should be injected during the period before the 20-th d after the last injection of chemopreparation, in case of radiation therapy - 12-15 d against the day of irradiation. In case of chemoradiation therapy AMSC should be injected after chemotherapy before the course of radiation therapy. This method enables to carry out chemoradiation therapy in patients with severe accompanying diseases that prevent such a therapy, and prevent the development of general toxic complications of chemoradiation therapy and medicinal and radiation-caused cystitis.
Biotransplant and method for treating the cases of osteoporosis / 2265442
Biotransplant has genetically unmodified mesenchyma stem cell culture as active component obtained from fetal donor autologous material. The tissue is subjected to disaggregation and the produced cell suspension is resuspended and cultivated on growth medium containing transferrin, insulin, fibroblast growth factor and heparin to accumulate mature stroma in cell culture. Method involves intravenously dropping mesenchyma stem cell culture in the amount of 50 to 500 mln in 50-100 ml of physiologic saline.
Biotransplant and method for treating the cases of diabetes mellitus of type i and ii / 2265443
Biotransplant has genetically unmodified mesenchyma stem cell culture as active component obtained from fetal donor autologous material and genetically unmodified fetal myoblast culture. Method involves intravenously dropping mesenchyma stem cells in the amount of 50 to 500 mln in 50-100 ml of physiologic saline. The fetal myoblast culture is intramuscularly introduced at a dose of 100 mln of cells per 10 kg.
Method for treating acute myocardial infarction at st segment lifting / 2266125
One should introduce the suspension of autologous mononuclear medullary cells without preliminary cultivation in to the mouth of coronary artery nourishing infarction area. Cell suspension should be introduced at the quantity of 100-150 mln. cells immediately after stenting coronary artery due to technique of passive passage. The procedure should be performed on the 14th - 21st d against the onset of the disease mentioned. The method provides efficient counterbalance of cardiomyocytes due to applying valuable stem cells at excluding complications induced by arterial occlusion.
Method for production of conditioning medium having colony-promoting activity / 2270022
CBA/CaLac intact mouse spleen is homogenized in medium containing 95 % of RPMI-1640 and 5 % of ethyl silicate; filtered through nylon grid; washed by centrifugation with RPMI-1640. at 1500 rpm for 10-15 min; final concentration of living splenocites is adjusted to 2x106 colony/ml using media containing 10 % of ethyl silicate, 1 % of L-glutamine; 0.2 % of gentamycine, 0.05 % of mercaptoethanol, 89 % of RPMI-1640; obtained product is cultivated at 37°C for 6 days, centrifuged and supernatant containing target product is separated followed by storage at -20°C. Before filtering 1 ml of distilled water is added in sterile spleen homogenate and after 25 sec 10 ml of RPMI-1640 medium is added. Target product is stored for 1 month or less.
Cell elimination by using viruses / 2270685
Invention relates to uses of viruses capable of reducing undesired cells in ex vivo mixtures of normal marrow or peripheral blood cells and tumor cells, such as leucosis or lymphoma cells due to interaction of abovementioned mixture with vesicular stomatitis virus. Invention also relates to method for malignant tumor treatment in mammalian. Claimed method includes sampling of mammalian marrow or peripheral blood cells; interaction of said cells ex vivo with vesicular stomatitis virus; mieloablative therapy; and transplantation of purified hematopoietic cells into mammalian. In another embodiment invention relates to method for malignant tumor treatment in mammalian having transplant of marrow or peripheral blood stem cells. Said method includes interaction ex vivo of collected transplant cell with vesicular stomatitis virus and administration of these purified cells in mammalian. Method of present invention makes it possible to reduce risk of malignant tumor backset in mammalian with transplanted hematopoietic cells.
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(57) Abstract: The invention relates to medicine, in particular, to a method for immunomodulator of mielopid. The proposed method is that cells in the bone marrow pigs are cultivated in a nutrient medium, separating the supernatant by centrifugation, the latter is subjected to ultrafiltration membranes with a threshold permeability 10.000 Yes at temperatures from +4°C to room temperature, then concentrated by lyophilization, hold gel chromatography on Sephadex G 25, selected fractions with molecular weight of 600 to 3000 D re-concentrated by lyophilization. The proposed method is compared with the known, allows you to get mielopid more immunostimulatory activity that entails a reduction in therapeutic doses of the drug and increasing the number of doses received from the same number of bone marrow cells. table 2. The invention relates to medicine, namely to immunology and can be used in the production of immunologically active drug mielopid. Described is a method of obtaining mielopid, formerly known as stimulator antitelomerase, by separating the cells from the bone marrow pigs, their cultivation in mittelhammer on column filled with Sephadex G-50. Faction, erwerbende in release of cytochrome G with a molecular weight of about 13000 Da, concentrated by lyophilization. Also known is a method of obtaining mielopid by culturing bone marrow cells in a nutrient medium, separating the supernatant by centrifugation, concentration, gelfiltration concentrated supernatant to a column of Sephadex G-25, the sampling fractions (mol.weight of 1000-2000 and desalting on a column of Sephadex C-10. The finished product is dried by lyophilization. The disadvantage of this method is a little out of the drug (from 1 kg nizatidine bones, 1,0.1010bone marrow cells receive 25-30 doses of the drug) and its low specific activity. Mielopid standardized so that 1 dose of drug contained 3 mg protein, determined by the method of Lowry. The stimulation index was not less than 1.6 at concentrations of 50 µg/ml. The purpose of the invention increased immunostimulatory activity mielopid. This objective is achieved in that the bone marrow cells of pigs are cultivated in a nutrient medium, the supernatant is separated by centrifugation, subjected to ultrafiltration membranes hold gel chromatography on Sephadex G-25, selected fractions with molecular weight of 600-3000 Da re-concentrated by lyophilization. Distinctive perelom of the method is that the selection of the target product are carried out by ultrafiltration on membranes with a threshold permeability of 10,000 Da at a temperature of +4aboutWith up to the room. In more detail, the method is carried out as follows. Bone marrow from the rib bones of pigs after their crushing wash by shaking in a thermostatted shaker at 37aboutC for 1 h in serum-free medium 199. Next, cells are washed twice by centrifugation and placed in a medium for culturing. Media for cultivation is based on 199 environment. The cultivation is carried out in a thermostatted shaker or roller installation at 37aboutC for 20 hours, the Supernatant is separated from the cell mass by centrifugation at a temperature of +4aboutWith speeds of 8,000 rpm for 20 min. The obtained culture supernatant of bone marrow cells subjected to ultrafiltration cassettes firm Millipor" threshold permeability 10000 Da at a temperature of +4aboutWith up to room temperature and the excess pressure of 0.5 ATM system. Received on o dissolved in sterile pyrogen-free water, centrifuged, the clarified concentrate is subjected to microfiltration, and then gel-chromatography on a column filled with Sephadex G-25, the selected fraction with a molecular mass of 600-3000 Da again subjected to lyophilization. From 1 kg of raw material (1,h10cells) receive 250-300 doses of the drug. Verification of the claimed technical solution for compliance with a criterion of "Inventive step" showed that neither the patent nor in scientific and technical literature not found in the set of features described in the claims. The following are specific examples of the execution of the method and analysis of the biological properties of mielopid. P R I m e R 1. 25 kg tipped rib bones of pigs crushed in sterile conditions. To homogenizate add 3 liters of sterile 199 environment. The suspension is shaken on a shaker at 37aboutC for 1 h to flush the bone marrow cells. Filtration separates containing cell supernatant volume of 3 L. the Cells are washed twice by centrifugation 199 medium at the temperature of +4aboutC. the resulting cell volume of 50 ml (15-25.1010) pour 25 l and 199 medium, bringing the cell concentration up to 1010cells/liter and incubated for 20 h at 37aboutaboutC. the Obtained filtrate volume 23 l freeze-dried at a temperature of (-30aboutC)-0aboutC. the Dry product is dissolved at +4aboutWith 350 ml of pyrogen-free sterile water for 30 min under vigorous stirring. Nerastvorim the precipitate was separated by centrifugation at 8000 rpm at +4aboutFor 20 minutes the Supernatant volume of 300 ml sterile filtered over a filter with a pore diameter of 0.2 μm and applied to a column of Sephadex G-25 size h see Gel chromatography is carried out in sterile conditions at +4aboutWith using of the eluate 10 mm phosphate buffer pH 7,00, selecting a fraction (mol. the mass corresponding to 2000 Da. The concentration of mielopid in the resulting eluate was adjusted to 0.1-0.2 mg/ml by adding sterile 10 mm phosphate buffer pH 7.0. The solution mielopid sterile filtered through a filter with a pore diameter of 0.2 μm, poured into vials of 3 ml and freeze-dried at a temperature of (-30)-0aboutC. The product yield is 300 doses of the drug with a protein content by Lowry 0.3-0.6 mg/dose. The coefficient of stimulation is thus 1,9. P R I m m e R 2. The method is carried out as described in example 1 but conducting the ultrafiltration on membranes with a threshold permeability 10000 Da when the stimulation of 1.8. P R I m e R 3. The effect of the stimulation of antibody productions estimate at the peak of the immune response to sheep erythrocytes in vitro, by adding the test material in the cell culture of lymph nodes obtained from mice on the fourth day after immunization. The stimulation coefficient calculated as the ratio of the number of antibody productive cells (AFC) in the culture with the test sample to the amount of the KLA in the control culture. Anticlockwise effect of mielopid by the well-known and proposed methods are given in table.1. From the data table.1 shows that mielopid obtained by the proposed method has a 100-fold greater specific activity. Anticlockwise effect of mielopid at different temperatures of the ultrafiltration process is shown in table.2. The process of ultrafiltration at a temperature above room temperature or below +4aboutWith leads to lower output and loss of activity of the drug. Mielopid obtained by the proposed method is 100 times more specific immunostimulirutuyu activity that can reduce the dose of an administered drug in the treatment and from the same number of bone marrow cells by the CSOs brain pigs, their incubation in a nutrient medium, separation of the supernatant, followed by separation of the target product, wherein the target product is optionally purified by ultrafiltration on membranes with a threshold permeability of 10,000 D and at a temperature from +4oWith up to the room and gelfiltration on Sephadex G-25 and selected fraction with molecular weight of 600 to 3,000 Da.
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