174 results about "Low molecular mass heparin" patented technology

Aiming at the conditions that heparin has severe bleeding side effects in clinical practice and clinical application thereof is restricted, the invention discloses a technique for producing ultra-low molecular heparin sodium (calcium). The technique comprises the following steps of: taking heparin sodium solution, adding sodium nitrite solution for cracking; adjusting the lysis buffer by using alkaline; absorbing impurities by using an anion-exchange column; washing for obtaining ultra-low molecular heparin calcium; carrying out filtration by using an ultrafiltration membrane and obtaining a precipitate by using alcohol; and after desalting, dehydration, re-precipitation, cooling and drying, obtaining a finished product of ultra-low molecular heparin calcium. The product has better and safer antithrombotic effect under low level of anticoagulation, and can be widely used for preventing and treating diseases such as deep vein thrombosis, pulmonary embolism, disseminated intravascular coagulation, and the like.

The invention relates to the field of cell engineering and particularly relates to a method for separating human adipose-derived stem cells from human adipose tissues. According to the method, by adding a small amount of low-molecular-weight heparin calcium into normal saline, a mixed solution is obtained and used as a cleaning fluid to clean the human adipose tissues, residual impurities such as red blood cells in the human adipose tissues can be well removed and thus human adipose-derived stem cells with relatively higher purity can be separated, the content of the parenchyma cell is significantly reduced, the differentiation induction of the human adipose-derived stem cells is facilitated, the consumption amounts of collagenase I, trypsase and a serum-free culture medium can be reduced and the cost of the process is also decreased. In addition, by centrifuging and digesting the human adipose tissues for 1 hour at a speed of 150r/min, the human adipose-derived stem cells with significantly better activity can be obtained.

The invention relates to a method for detecting low-molecular-weight heparin by combining ion pair reversed phase chronmatogaphy and mass spectrum. The method comprises the steps of: detecting all compositions of low-molecular-weight heparin by combining ion pair reversed phase chronmatogaphy and high-resolution mass spectrum, separating main compositions from a sample through the ion pair reversed phase chronmatogaphy, obtaining precise molecular weight by the high-resolution mass spectrum, and calculating information of a carbohydrate chain sequence, including structures of two ends, length of a carbohydrate chain and substitution number of ethanoyl and sulfate, thus carrying out fine surface features on the low-molecular-weight heparin. The method has great practical values on improving the heparin detection level and ensuring drug safety.

The invention discloses a process for preparing high-purity low-molecular heparin sodium and belongs to the field of bioengineering. The process mainly comprises the following steps of: salting-out and impurity removal, supplementary material warming and oxidization, gel elution, ion exchange, ethanol precipitation, vacuum drying and the like. The problems that the existing heparin sodium structure analog is hard to remove, the purity can not achieve the requirements for medicinal use, the preparation process is complicated, and the like are solved. The process has the beneficial effects that by adopting a supplementary material warming and oxidization method, the preparation and the decolorization of the low-molecular heparin sodium are realized, and the heparin sodium structure analogue is effectively removed by using weakly-basic anion exchange resin; and the process has the advantages of shortened production period, saved cost, simple process, easiness in operation and is suitable for industrial production.

The preparation process of low molecular weight heparin calcium includes the following steps: dissolving refined heparin sodium in acetic acid aqua and adding sodium nitrite to the solution for cracking for 4-6 hr to obtain cracked solution; neutralizing the solution with calcium hydroxide to pH 5.5-6.0; filtering to eliminate insoluble matter, precipitating the solution with saturated ethanol solution of calcium chloride and collecting the precipitate; dissolving the precipitate in injection water and regulate pH with hydrochloric acid to 0.3-1.0; neutralizing the solution to pH 6.0-6.5; filtering to eliminate insoluble matter, precipitating the filtrate with saturated ethanol solution of calcium chloride and collecting the precipitate; and conventional decompression drying to obtain low molecular weight heparin calcium product. The product has average molecular weight of 3000-5000 D, Xa resisting activity not lower than 90 IU/mg, Xa resistance/IIa resistance ratio of 3-5 and nitrite content lower than 2 ppm.

The invention provides a freeze-dried preparation for treating colpitis. The preparation includes an antimicrobial peptide freeze-dried powder and an antibacterial peptide basic liquid. The antibacterial peptide freeze-dried powder comprises the following components according to the total mass percentage: 0.1%-8% of an antibacterial peptide human beta defensin, 0.01%-0.2% of a low molecular weight heparin sodium and 1%-15% of a stabilizing agent; the antibacterial peptide basic liquid comprises the following components according to the total mass percentage: 0.01%-0.4% of hyaluronic acid, 0.5%-4% of menthoxypropanediol, 0.5%-5% of a honey extract, 0.5%-5% of PCA sodium, 0.5%-5% of dissolved protease, 0.3%-0.7% of 1,2 hexanediol, 1.5%-6% of oat beta dextran, 1%-5% soybean isoflavone, and the balance of deionized water or water for injection. The antimicrobial peptide freeze-dried preparation provided by the invention has broad-spectrum antibacterial activity, and strong killing effect on bacteria, fungi, viruses, protozoa and cancer cells, and even can enhance immunity, accelerate wound healing process; in addition, the preparation has obvious curative effect, and no rebound or side effect.

The invention relates to a reverse-phase chromatography and mass-spectrometry combined detection method for a complete low-molecular-heparin degradation product through precolumn derivatization. All components of the complete low-molecular-heparin degradation product are detected by combining reversed-phase chromatography and a high-resolution mass spectrum; all the components in a sample are separated through the reversed-phase chromatography, and a precision molecular weight can be obtained through the high-resolution mass spectrum; besides detection of eight common heparin disaccharides, tail end modification structures and special structures such as 3-O-tetrose-hydrosulfates relevant to anticoagulation and trisaccharides obtained through peeling reaction can be also detected; therefore, the structure of each complete low-molecular-heparin degradation product can be precisely represented. According to the reverse-phase chromatography and mass-spectrometry combined detection method, the problem that only the eight common heparin disaccharides can be detected in the prior art is solved; the reverse-phase chromatography and mass-spectrometry combined detection method has an extremely large practical value in research, development and production control of low-molecular-heparin imitating drugs and guarantee of the safety of the drugs.

The invention discloses a process for producing low molecular sodium heparin by using raw sodium heparin as a raw material, which comprises the steps of: producing a sodium heparin solution; degradingthe solution; neutralizing the solution; collecting a precipitation; producing low molecular raw sodium heparin; producing low molecular fine sodium heparin and the like. The low molecular sodium heparin has the characteristics of rich sources of the raw material, high yield, stable and reliable quality, high purity, low cost, long curative effect, no toxic or side effect, simple process, convenient operation and no 'three wastes' discharge, has the effects of coagulation resistance, thrombosis resistance, tumor resistance, inflammation resistance, allergy resistance and the like and the efficiency of regulating blood lipid, has significant curative effect and wide application range, can be produced into injections, capsules, ointments and the like, is convenient for massive production, and is widely used for producing coagulation resistant low molecular sodium heparin.

The invention discloses a production method of dalteparin sodium. According to the production method, a heparin sodium coarse product is taken as a raw material, a low-molecular-weight heparin sodium coarse product is obtained through degradation, reduction and ethanol precipitation; compared with a method taking a heparin sodium fine product as the raw material, the method is wider in application range and can better control the quality of a dalteparin sodium fine product from the source. Innovatively, molecular weight and distribution of the low-molecular-weight heparin sodium coarse product before chromatography are tested through HPLC (high performance liquid chromatography), corresponding required-to-be-removed areas of macromolecules and micromolecules and corresponding volumes are calculated according to the formula, filtrate in the chromatography process is collected, a low-molecular-weight heparin sodium fine product liquid which meets the requirements is accurately and quantitatively obtained, and use of an expensive molecular weight interception filtering membrane is avoided; a one-pot process is achieved through oxidation, sterilization filtration and sediment dehydration, the operation is simplified, and time and labor cost are saved; and a new method for preparing a raw dalteparin sodium material is developed on the basis of the prior art, and industrial production of dalteparin sodium is realized.

The invention relates to a preparation process of nadroparin calcium. The preparation process comprises the following steps: S1, degrading heparin sodium; S2, reducing degradation liquid; S3, performing ultraviolet radiation, namely radiating reduction liquid by using an ultraviolet lamp to remove residual nitro compounds from the reduction liquid and reduce the content of N-NO groups; S4, performing calcium conversion and concentration, namely performing ultrafiltration on an ultrafiltration membrane by using a 5% calcium chloride solution, and concentrating the reduction liquid until the solid-liquid ratio is 1:10; S5, performing anion exchange chromatography, namely performing chromatography by adopting anion exchange resin, and collecting elution liquid; S6, freeze-drying to obtain nadroparin calcium. According to the process, the molecular weight of low-molecular-weight heparin calcium is controlled through the anion exchange chromatography, so that the molecular weight can be accurately controlled within 1% and the yield can be increased to more than 50%; moreover, the automation degree is high and the operation is convenient, so that a powerful guarantee is provided for increasing the production capacity.

The present invention relates to the extraction and purification method of low molecular weight heparin for treating blood coagulation and thrombus. The coarse low molecular weight heparin material is dissolved in NaCl solution and chromatographically separated with molecular sieve gel layer to obtain the chromatographic separated liquid, and the chromatographic separated liquid is alcohol precipitated, dewatered and dried to obtain low molecular weight heparin segment. Before chromatography, the coarse product is irradiated with ultraviolet lamp or reacted with oxidant sodium hypochlorite or hydrobromic acid to eliminate harmful ¿CN-NO- radical, and treated with strong anionic exchange resin to eliminate methanol and other chemical impurity. The present invention has the low molecular weight heparin product reaching relevant international standard, and the method is simple and high in yield.

The invention relates to an engineering strain construction method for improving the activity of heparinase I. An amino acid sequence is optimized according to spatial structure analysis on the heparinase I to obtain Hep169 with the activity improved by 48% compared with the heparinase; a Hep169 gene is optimized according to codon preference, a gene DNA is obtained through artificial synthesis and is cloned into an expression vector to perform fusion expression with SUMO and other labels, host cell conversion and screening are performed to establish a soluble genetic engineering production system of the Hep169, analysis results show that a target protein obtains an efficient soluble expression, has very good biological activity and can efficiently crack heparin to generate low-molecular-weight heparin. By the adoption of the method, the highly active HepI is provided, a new method is provided for efficient soluble expression gene engineering production of the heparinase I, the production cost of the low-molecular-weight heparin and other drugs can be effectively reduced, and the engineering strain construction method has wide application prospect.

The invention relates to a preparation method for low-molecular heparin originated from a new species. The preparation method comprises the following steps: (1) fully dissolving heparin sodium prepared from raw materials bovine lungs, ox intestines and/or goat intestines in water, and adding a benzethonium chloride solution to prepare heparin benzethonium chloride salt; (2) dissolving the heparin benzethonium chloride salt in dichloromethane, adding benzyl chloride, stirring the mixture to react, reducing the temperature and adding a sodium acetate methanol solution to prepare heparin benzyl ester; (3) dissolving the heparin benzyl ester in water, adding sodium hydroxide, reducing the temperature after reaction, adjusting the pH to neutral, adding sodium chloride, and then adding alcohol to prepare an enoxaparin sodium crude product; and (4) dissolving the enoxaparin sodium coarse product in water, and purifying and drying the mixture to obtain enoxaparin sodium. According to the preparation method provided by the invention, the condition that the low-molecular heparin can be obtained by heparin sodium prepared from bovine lungs, ox intestines and goat intestines is found for the first time, and the prepared low-molecular heparin is lower in molecular weight compared with that of existing low-molecular heparin, so that the biological activity is higher and the low-molecular heparin has a wide market application prospect.

The invention relates to an ion pair antiphase chromatography and mass spectrometry combined detection method for low-molecule heparin part degradation products. All components of the low-molecule heparin part degradation products are detected in a manner of combining ion pair antiphase chromatography and high-resolution mass spectrometer; all the components in a sample are separated through the ion pair antiphase chromatography, and a precise molecule weight is obtained through the high-resolution mass spectrometer, so that sugar chain sequence information, including structures of two ends, lengths of sugar chains and substitution numbers of acetyl and sulfuric acid, of all the components is calculated, so that the structures of the low-molecule heparin part degradation products can be finely represented. The ion pair antiphase chromatography and mass spectrometry combined detection method can detect the structures of the two ends, the lengths of the sugar chains and the substitution numbers of the acetyl and the sulfuric acid of the low-molecule heparin part degradation products; the problem in the prior art that only an ultraviolet absorption atlas for degradation of low-molecule heparin parts can be obtained is solved; the ion pair antiphase chromatography and mass spectrometry combined detection method has a high practicability value in improvement of the detection level of heparin and guarantee of drug safety.

The invention discloses a process for preparing heparin sodium through membrane separation. The process comprises the steps: intestinal mucosa enzymolysis, ceramic membrane filtration, ion exchange column chromatography, nanofiltraiton membrane concentration, alcohol precipitation and drying, wherein the optimal condition for the intestinal mucosa enzymolysis is formed by the proper proportion of intestinal mucosa to water, enzymolysis temperature and pH value of a system. A protease can take the optimal enzymolysis effect, the structure and activity of heparin are not damaged when the glucosidic bond linked between protein and the heparin is hydrolyzed, meanwhile, fewer low-molecular heparins are generated, the activity of the heparin can be improved by 20%, and the generated low-molecular heparins can be reduced by 10%. Due to the adoption of the ceramic membrane filtration, protein impurities and the heparin in an enzymatic hydrolysate can be effectively separated, so that the heparin loss is greatly reduced and can be reduced by 5-8% as comparison with that of the traditional process, the production cost is reduced, the discharged wastewater can be reduced by 50-60%, the resin utilization ratio can be increased by 40-50%, the consumption of auxiliary materials is low, the quality of the heparin sodium is stable, the process is convenient to operate, and the defects such as high impurity content in the enzymatic hydrolysate, serious waste liquid discharge pollution, high personnel utilization rate and the like are overcome.

The invention belongs to the technical field of medicines and relates to a compound low-molecular heparin sodium liposome gel and a preparation method and application thereof. The compound low-molecular heparin sodium liposome gel comprises low-molecular heparin sodium, asiaticoside, lipid components, a membrane softening agent and gel matrix components, wherein the low-molecular heparin sodium and asiaticoside are pharmacological active ingredients, and based on the total gel of 100g, the content of the low-molecular heparin sodium is 5000-35000IU, and the content of the asiaticoside is 0.1-2.5g. The preparation method comprises the following steps of: (1) preparing a compound low-molecular heparin sodium liposome by employing an ethanol injection-ultrasonic method or ethanol injection-high pressure homogenizer method; (2) preparing a blank gel; (3) adding the compound low-molecular heparin sodium liposome into the blank gel, uniformly stirring, and obtaining the compound low-molecular heparin sodium liposome gel. The gel is mainly used for treating skin injury and scar.

The invention relates to the technical field of stem cells, in particular to a mesenchymal stem cell injection as well as a preparation method and application thereof. The mesenchymal stem cell injection comprises mesenchymal stem cells, human serum albumins, low-molecular weight heparin calcium, vitamin C, lentinan, dimethyl sulfoxide and a menstruum. The mesenchymal stem cell injection provided by the invention can effectively maintain the activity of the stem cells, can increase the content of striatal dopamine and can effectively treat the Parkinson's disease.

The invention relates to a preparation method of a novel source of low molecular weight heparin from nitrous acid degradation. The steps are as follows: (I) completely dissolving raw materials in water, adding sodium nitrite to prepare a reaction solution A; (II), adding sodium borohydride, and reacting to prepare a reaction solution B; (III) adding an alcohol solution, conducting solid-liquid separation, drying to prepare a solid A, and then dissolving the solid A in water to prepare a reaction solution C; (IV) replacing sodium ions with calcium ions by cation resin exchange to prepare a reaction solution D; and (V) adding alcohol solution to the reaction solution D, fully mixing, conducting solid-liquid separation, purifying, and drying to prepare the low molecular weight heparin. The invention can fundamentally promote the diversification of species sources for low molecular heparin, expand the supply of raw materials for heparin drugs, and achieve a higher degree of stabilization of the market of heparin and low molecular weight heparin drugs.

The invention relates to the application of pioglitazone and heparin or low molecular heparin or pharmaceutically acceptable salts thereof or derivatives thereof to preparing a pharmaceutical composition, in particular to the application to preparing a medicament for preventing or treating fatty liver. the purpose of the pioglitazone and the heparin or the low molecular heparin or the pharmaceutically acceptable salts thereof in lessening fat deposition in rat hepatocytes and the synergy of a pioglitazone and heparin or low molecular heparin compound in preventing or treating the rat fatty liver are proved by animal tests. The invention provides a new candidate medicament for searching a medicament for lessening fat deposition in hepatocytes and treating diseases caused by clinic lipid metabolism, such as fatty liver, and the like and therefore enriches the prior art.

The invention relates to the technical field of stem cells and especially relates to an umbilical cord mesenchymal stem cell injection liquid, a preparation method and an application thereof. The injection liquid comprises umbilical cord mesenchymal stem cells, human albumin, low molecular heparin calcium, vitamin C, panax japonicus majoris saponin and a solvent. The injection liquid is prepared from the umbilical cord mesenchymal stem cells, so that the materials are convenient to obtain and the method does not violate ethical issue. The injection liquid can effectively prolong preservative time of the cells and the preserved stem cells have high activity, so that the injection liquid is convenient to use. After refusion, the injection can promote differentiation and maturation of the mesenchymal stem cell to liver cells, thus reducing incidence rate of GVHD.

The invention discloses an umbilical cord mesenchymal stem cell preserving fluid. The umbilical cord mesenchymal stem cell preserving fluid comprising the following components at different concentrations: low-sugar DMEM culture medium of 5 to 10% (V/V), human serum albumin of 15 to 25g/L, sodium chloride of 8 to 10g/L, low-molecular heparin sodium of 2000 to 3000AXaIU/mL, glucose of 20 to 50g/L, vitamin C of 100 to 150mg/L, and salidroside of 10 to 30mu M. The umbilical cord mesenchymal stem cell preserving fluid can effectively preserve the umbilical cord mesenchymal stem cells under the condition of 4 DEG C and keep stable cell phenotypes. All the components of the umbilical cord mesenchymal stem cell preserving fluid are safe and non-toxic to a human body, subsequent use of the umbilical cord mesenchymal stem cells is facilitated, and the umbilical cord mesenchymal stem cell preserving fluid is very suitable for preservation and transportation of the umbilical cord mesenchymal stemcells and has a wide application prospect.