The way to obtain therapeutic agents that regulate cell differentiation
(57) Abstract:The invention relates to biotechnology and can be used in pharmacology and medicine. The method consists in homogenizing the muscle tissues of the embryo of the animal, after which the homogenate was incubated for 10-20 h, the extraction is carried out, and then separating the precipitate, and the resulting solution was further diluted with saline to achieve a concentration of protein in it 15-25 mg/ml, add the preservative, separating the resulting suspension, a solution was incubated for 48 h, sterilized, termostatic it stand 30-40 days at a temperature not exceeding 15oC remove the resulting suspension and the solution is subjected to interaction with immobilized proteolytic enzyme. The technical result is obtained a highly effective tool in the treatment of diseases associated with impaired differentiation of cells. 1 Il. The invention relates to biotechnology and can be used in pharmacology and medicine.Differentiation of cells occurs in various physiological processes in the body and primarily with violations functions of the immune system and the consequences of this naree closest to the claimed solution is the method of obtaining remedies regulating the differentiation of cells and intended for the treatment and prevention of cancer (French Patent N 2451193, 1979).The known method consists in grinding the embryos of animals, their homogenization in physiological solution, obtaining an extract of embryonic complex substances, and separating the resulting precipitate by filtration, then the filtrate add an antibiotic. As a result of implementation of the specified sequence of actions receive drug treatment full embryonic complex. The treatment received by the drug is in the necessary course of injections.The prototype method is quite simple in execution, but it cannot provide a highly effective drug, as no cleavage of macromolecular precursors and activation of regulatory peptides. The resulting preparation contains the formed elements and high molecular weight proteins that largely determines its side effects, manifested in allergies and in her terrible complication is anaphylactic shock.The present invention is to provide a method for obtaining a therapeutic agent that regulates the differentiation of cells, to which their protein-peptide components and neutralizing other, provides high efficiency of the final product and excludes its side effects.The problem is solved in that in a method of producing therapeutic agents that regulate the differentiation of cells, which consists in grinding the embryonic tissues of the animal, their homogenization and extraction of physiological solution, separating the precipitate from the solution, according to the invention as embryonic tissues take the muscle tissue of the animal, the homogenate was incubated for 10 to 20 hours, and after extraction, the treatment solution is performed within 24 0.5 hours, then after separation of the resulting precipitate the resulting solution was further diluted with saline to achieve a concentration of protein in it 15 - 25 mg/ml and add to it the preservative in an amount not less than 0.05%, after separation of the resulting suspension solution was incubated for 48 hours, sterilized and thermostatic it for at least 3 days at a temperature of 36oC, and the solution was incubated 30 - 40 days at a temperature not exceeding 15oC, and remove the resulting suspension, and the solution is subjected to interaction with immobilized proteolytic enzyme, with the mode specified Sanogo method includes especially the protein-peptide complex (BOD), which is an instrument with a certain specificity and required quality.Use as raw material embryonic muscle soft tissue of the animal is responsible for the specificity of the obtained funds. Keeping these tissues homogenate for 10 to 20 hours due to biosynthesis extreme proteins, the nature of which depends on the quality of therapeutic agent. If more time is specified, the deterioration of the quality of the product, because formed necrotic proteins that cause toxicosis. With less time also receive the drug is of low quality.After extraction BOD in saline solution proposed to stand the solution for 24 hours under these conditions, the maximum weight of the extraction, the extract solution outside the specified time dramatically changes the quality of the final product. In the process of aging are formed predecessors active proteins, which determine the specificity of the drug.At this stage of the biosynthesis of a precipitate of insoluble proteins (tissue, plasma and other), they are separated, and the resulting solution was further diluted with saline to achieve the iza.Adding to the obtained solution of the preservative in an amount not less than 0.05% is intended for antimicrobial, antifungal, etc., disinfection of the final product.The formed suspension, representing proteins, membranes, structural proteins, lipid particles and other, separate, since the subsequent process of autolysis will apply to them, that adversely affects the quality of therapeutic agent.The remaining solution was incubated for 48 hours. Experimentally proved, that this is enough time to complete the processes of aggregation of particles. The resulting solution is sterilized to remove viruses, bacteria, mantles, fungi, etc., Sterilization can be performed by any one adopted in this field by the method in accordance with who requirements.The following actions are obtained by solution of the BOD are to autolysis. The temperature of the solution is carried out for not less than 3 days at a temperature of 36 2oC.The timing of incubation at this temperature due to reaching the maximum specificity obtained a remedy.The processes at this stage, can be opredelyaemye the drug is non-immunogenic.Further keeping the solution within 30 - 40 days at a temperature not exceeding 15oC is determined by the following factors under these conditions, gently ends the process of autolysis with simultaneous aggregation of the formed particles that separate.The remaining BOD solution is subjected to proteolysis by immobilized proteolytic enzyme, for example, on a column filled them.The invention is illustrated by specific example.To obtain therapeutic agents that regulate cell differentiation, embryonic take the muscle tissue of cattle. Selected raw materials are crushed, homogenized at a temperature not exceeding 8oC, the homogenate was incubated for 16 hours, diluted with chilled saline solution, then after extraction solution further maintained at a temperature of 4 - 8oC within 24 hours.In the process of keeping a precipitate, which was separated by centrifugation at a speed of 3000 rpm. within 1 hour at a temperature of 2 to 4oC.The resulting solution was diluted with saline to achieve a concentration of protein in it 21 mg/ml, is added dropwise a preservative which means passing the solution through a membrane filter with a pore diameter of 0.45 μm and incubated filtrate within 48 hours.After that make a sterilization solution, for example, by passing it under aseptic conditions through a membrane filter with a pore size of 0.22 μm. Thus obtained a sterile solution hermetically closed and thermostatic for 3 to 4 days at a temperature of 36oC. After incubation the solution incubated for 35 days at a temperature not exceeding 15oC, the resulting suspension was separated by centrifugation, and the filtrate is optionally passed through a filter with pore size 0.45 µm.The resulting solution BOD passed through the column (at a temperature of 25 - 30oC) filled with agarose 4B, the immobilized enzyme, such as trypsin.The rate of elution solution BOD set on the basis of matching electrophoregram of the eluate with the control electrophoregrams BOD.The resulting liquid aseptically sterilized by passing it through a membrane filter with a pore diameter of 0.22 μm, and then poured into ampoules and sealed.The resulting tool is a transparent liquid with light yellow color without a smell, which can be mixed with water or saline solution in any proportion. the oC - 1,3410 of 0.0004, pH 6,5 - 7,0.The drawing shows a control electrophoregram received the drug.Thus, in the inventive method obtained a therapeutic agent that regulates the differentiation of cells ("Probes") that is formstate and which is widely tested on patients. The tool is recognized as highly effective in the treatment of diseases associated with impaired differentiation of cells. The way to obtain therapeutic agents that regulate the differentiation of cells, which consists in grinding the embryonic tissues of the animal, their homogenization and extraction of physiological solution, separating the precipitate from the solution, characterized in that as embryonic tissues take the muscle tissue of the animal, the homogenate was incubated for 10 to 20 h, and after extraction, the treatment solution is performed within 24 0.5 h, then after separation of the resulting precipitate the resulting solution was further diluted with saline to achieve a concentration of protein in it 15 - 25 mg/ml and add to it the preservative in an amount not less than 0.05%, after separation of the resulting suspension solution was incubated for 48 h, female sterilization is the temperature, not exceeding 15oC, and remove the resulting suspension, and the solution is subjected to interaction with immobilized proteolytic enzyme, and the mode of the specified interaction set on the basis of a control sample obtained funds.
FIELD: medicine, otorhinolaryngological surgery.
SUBSTANCE: one should apply thin layer of "Solcoseryl" gel onto osseous facial walls of frontal and maxillary sinuses at the border with trepanation opening after removing pathological content out of them and before applying a transplant out of flat bone of human fetal cranial arch that exceeds the diameter of trepanation opening by 3-4 mm. Then, one should additionally fix the transplant by affecting with distal edge part of a light guide of semi-conductor laser "ATKUS-15" with contact-type technique at output power of laser radiation being 8 W at constant mode. The method enables to increase fixation density of allobrefobone to osseous walls of sinus along its whole diameter.
EFFECT: higher efficiency of fixation.
FIELD: medicine, surgery.
SUBSTANCE: the present innovation deals with local treatment of erosive-ulcerous defects of skin and mucosa, inflammatory intestinal diseases and prolongly non healing wounds, among them. The method includes the use of diploid embryonic fibroblasts cultivated beyond the body, moreover, the fibroblasts should be pre-affected with dexametason at concentration of 10-4 M/l under culture conditions for 1 h followed by washing it from preparation and changing the medium. Then one should apply the developed supernatant in the form of local application onto affected area once daily. The method enables to decrease inflammatory manifestations in wounds and erosive-ulcerous defects of skin and mucosa, stimulates regenerative processes at minimal restrictions while applying the present innovation from the side of lesion section and decreases financial expenses.
EFFECT: higher efficiency of local treatment.
7 dwg, 2 ex, 6 tbl
FIELD: veterinary science.
SUBSTANCE: one should apply a honey-tissue preparation that contain emulsion out of the walls of pregnant uterus and ovaries without cow's yellow bodies 6.0 ml, natural bee honey 5.0 g, isotonic sodium chloride solution 2.0 ml sodium benzoate caffeine 1.5 g to be introduced into dorsal lumbar muscles both from the right and from the left per a half of the dosage being equal to 0.03-0.04 ml/kg either once or twice at 6-7-d-long interval. The present innovation enables to improve metabolism, normalize the work of hormonal and nervous systems, normalize functions of uterine muscles at hypo- and atonias, at delayed afterbirth and restore affected ovarian functions.
EFFECT: higher efficiency of therapy.
FIELD: veterinary medicine.
SUBSTANCE: the present innovation deals with methods to activate the activity of protective mechanisms and organs of hormonal regulations. One should introduce a tissue preparation for an animal, moreover, to obtain it is necessary to apply 2.5 g pregnant or postpartum uterus, per 1.5 g thyroid, parathyroid and thymus glands, 2.0 g pancreas, 2.5 g liver all purified against spare tissues from clinically and hematologically healthy animals from cattle group up to 4-6-yr-aged period. The organs mentioned should be reduced, then tissue mixture obtained should be mixed with fresh running water at 1:2 ratio to obtain emulsion to be thermally treated in water bath at 58-59 C. After cooling emulsion should be supplemented with formalin, ACD f-2 and natural bee honey to obtain the following ratio of components in ready-to-use product: tissue emulsion 10 g, 68.7%; ACD f-2 1.5 g, 10.3%; formalin 0.006 g, 0.4%; natural bee honey 3.0 g, 20.6%. Before being introduced for an animal one should carefully mix preparation to detect its dosage at 0.02-0.5 ml/kg animal body weight to be then introduced per Ѕ parts from the right and from the left into dorsal lumbar muscles either once or twice at interval of 7-9 d. The method enables to perform complex biostimulating and hormonal impact upon animal body due to keeping active substances of tissue preparation being obtained due to above-mentioned technique.
EFFECT: higher efficiency of prophylaxis and therapy.
FIELD: biopharmacology, medicine.
SUBSTANCE: the suggested biotransplant contains an active component: the culture of genetically unmodified neuronal stem cells (NSC) obtained out of anterior cerebral tissue of human embryos of the 1st trimester of pregnancy or periventricular cerebral area of 15-20 wk gestation and selectively multiplied under cultivating conditions up to the quantity of 10 (7) - 10 (9) pluripotent undifferentiated cells in composition of neurospheres, at the content of NSC being 50-500 mln., and 2 ml physiological solution. The method for treating ischemic insult deals with introduction of 50-500 mln. NSC in 2 ml physiological solution. The suggested biotransplant and method for treating ischemic insult enable to quickly restore and improve cerebral functions.
EFFECT: shortened terms of therapy.
5 cl, 2 ex, 1 tbl
SUBSTANCE: biotransplant has active ingredient like culture of genetically unmodified neuron trunk cells taken from anterior brain of the first pregnancy trimester human embryo or periventricular region of brain of 15-20 gestation weeks and selectively reproduced under cultivation conditions to the amount of 107-109 pluripotent non-differentiated cells in neurospheres having 50-500 mln of neuron trunk cells and 2 ml of physiologic saline. Method involves introducing 50-500 mln of neuron trunk cells and 2 ml of physiologic saline in one stage in endolumbal way.
EFFECT: enhanced effectiveness of treatment; accelerated repair and improvement of injured brain functions; stable treatment results.
5 cl, 1 tbl
FIELD: medicine, gerontology.
SUBSTANCE: beforehand one should detect human biological age and the age of human immune system to compare them with human calendar age: in case of predominance of at least one of them against calendar age one should once introduce the suspension of human fetal tissues into subcutaneous fiber of anterior abdominal wall. For this purpose one should apply fetal tissues being heterogeneous by their histogenesis and, also, preparations of placental origin. The innovation provides normalization of immunocytogram values and, thus, similarity of calendar and biological age for 1 yr.
EFFECT: higher efficiency of correction.
FIELD: medicine; medical engineering.
SUBSTANCE: 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.
EFFECT: accelerated recovery of bone tissue; positive biochemical factors dynamics; improved patient locomotor activity.
FIELD: medicine; medical engineering.
SUBSTANCE: 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.
EFFECT: enhanced effectiveness in repairing incretory function of pancreas and reducing resistance to insulin; reduced risk of nephropathy and other complications.
FIELD: biopharmacology, medicine.
SUBSTANCE: the present innovation deals with preparing the culture of genetically unmodified chondroprogenitor cells and method for treating parodontium diseases. Biotransplants, methods for their obtaining and method for treating parodontium diseases deal with introducing the suspension of mesenchymal stem and chondroprogenitor cells or their combination that contains 1-50 mln. cells/ml, intraligamentally from lateral and medial sides of every tooth and subperiosteally, into mandibular and maxillary parodontal parts.
EFFECT: higher efficiency of therapy.
13 cl, 1 ex