68 results about "Anticoagulant activity" patented technology

The invention relates to the field of establishment of fingerprints of traditional Chinese medicinal materials, in particular to an establishment method and application of a salvia miltiorrhiza polysaccharide HPLC (High Performance Liquid Chromatography) fingerprint. The establishment method comprises the following steps: preparation of a test solution: taking a salvia miltiorrhiza polysaccharide hydrolysis solution, carrying out derivatization in a reaction bottle, and passing through a 0.1-0.3 [mu] m microporous filter membrane; preparing a reference substance solution, namely weighing mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose and arabinose reference substances, and preparing into a mixed reference substance solution; performing HPLC (high performance liquid chromatography) determination; and establishing the salvia miltiorrhiza polysaccharide HPLC fingerprint spectrum. According to the method, the difference of monosaccharide composition and content in salvia miltiorrhiza medicinal materials from different producing areas and the spectrum-effect relationship between antioxidant activity and anticoagulant activity are clarified, and a certain reference basis is provided for quality control and evaluation of the salvia miltiorrhiza medicinal materials from the aspect of polysaccharide.

The invention relates to a monoclonal antibody with anticoagulant activity and application thereof, the antibody or fragment comprises a light chain variable region and a heavy chain variable region, and the amino acid sequence of the light chain variable region of the antibody or fragment is as shown in SEQ ID NO: 1; the amino acid sequence of the variable region of the heavy chain is as shown in SEQ ID NO: 3. The monoclonal antibody disclosed by the invention has the effect of inhibiting the activity of a co-coagulation pathway in a coagulation cascade reaction, and can be applied to prevention and treatment of thrombotic diseases.

The invention discloses a fusion protein rHTA and application thereof, the fusion protein rHTA is sequentially composed of a Hirudin protein, a thrombin cleavage site and an AnxV protein from the N terminal to the C terminal, and the amino acid sequence is as shown in SEQ ID NO: 4 from the 1st site to the 393th site from the N terminal. Experiments prove that the fusion protein rHTA is specifically activated by thrombin at the local part of thrombus and releases AnxV protein and Hirudin protein, and the AnxV protein can specifically recognize and combine with PS, so that the fusion protein rHTA is enriched at the thrombosis part containing platelets in a targeted manner, the Hirudin protein is locally released under the action of thrombin, local precise anticoagulation is realized, and the anticoagulation efficiency is further enhanced. Therefore, the fusion protein rHTA provided by the invention not only has high anticoagulant activity, but also realizes enrichment and release at the thrombus formation part, and reduces the bleeding risk. The method has an important application value.

The invention discloses a preparation method of a fucoidan oligomer, which realizes efficient degradation of fucoidan under specific conditions by strictly controlling the addition amount of TEMPO, sodium bromide and sodium hypochlorite. Specifically, the ratio of the addition amount of TEMPO to the mass of fucoidan sulfate is 0.5-5%, the ratio of sodium bromide to fucoidan sulfate is 5-40%, the use amount ratio of fucoidan sulfate to sodium hypochlorite is 1 g: (0.1-10) mmoL, strong acid or enzyme preparations do not need to be added, and auxiliary means such as microwaves or ultrasound are not depended on, so that the preparation process is simple, and the preparation cost is low. The oligomer with the average molecular weight lower than 80 kDa can be obtained within one hour, the oligomer with the average molecular weight about 20 kDa can be obtained within four hours, and meanwhile the content of sulfate groups is kept to be about 25%. The functional groups of the fucoidan are hardly damaged, and the prepared oligomer has a remarkable effect in the aspect of in-vitro anticoagulant activity.

The invention belongs to the technical field of biochemistry, and particularly relates to an efficient enzymatic synthesis method of holothurian glycosaminoglycan. According to the invention, a high-efficiency enzymatic synthesis approach of the sea cucumber glycosaminoglycan is established, sulfated fucose and chondroitin sulfate are used as raw materials, and the sea cucumber glycosaminoglycan with anticoagulant activity is efficiently prepared by using the alpha 1, 3-fucosyltransferase mutant and L-fucose kinase / GDP-fucose pyrophosphorylase and adopting a one-pot multi-enzyme synthesis strategy. And important technical support is provided for development of novel anticoagulant drugs.

To provide clinical anticoagulants that are advantageous for use as surgical anticoagulants and are of sufficiently high molecular weight to greatly reduce rapid renal clearance.SOLUTION: The present disclosure provides a single-stranded nucleic acid molecule that includes at least one A-form double-helical structure and at least one crossover region, at least one kissing-loop region, and at least one nucleic acid aptamer having anticoagulant activity. The present disclosure also provides nucleic acid molecules comprising one or more aptamers having anticoagulant activity, as well as corresponding nucleic acid antidotes, for modulation of blood coagulation relating to disease and surgical intervention.SELECTED DRAWING: Figure 1

The present invention relates to an anticoagulant heparin-derived oligosaccharide biphenyl dimer, and a preparation method therefor and a use thereof. The anticoagulant heparin-derived oligosaccharide biphenyl dimer has a structure as represented by general formula I, and a benzene ring can be replaced with substituted phenyl, an aromatic heterocyclic ring or a substituted aromatic heterocyclic ring. The heparin-derived oligosaccharide biphenyl dimer of the present invention has few synthesis steps and a significantly high overall yield, has antithrombin-dependent potent and specific anti-Xa factor activity and no obvious anti-IIa activity, anticoagulant activity being efficiently neutralized by a protamine, and has excellent pharmacokinetics characteristics. Therefore, the present invention is suitable for preparing novel anticoagulant and antithrombotic drugs having cost advantages, and high quality and safety.

The invention provides a fucosan sulfate compound as well as a preparation method and application thereof, and belongs to the technical field of organic chemistry and medicinal chemistry. The anticoagulant activity of the fucosan sulfate compound is high, the anticoagulant activity of the compound 29, the anticoagulant activity of the compound 30, the anticoagulant activity of the compound 37 and the anticoagulant activity of the compound 58 are high, EC2.0 * multiplied by APTT reaches 13.6 mu g / mL, 14.5 mu g / mL, 16.4 mu g / mL and 12.9 mu g / mL respectively, and the activity of the EC2.0 * multiplied by APTT is close to that of an anticoagulant drug enoxaparin commonly used in clinic. However, different from enoxaparin acting on a common blood coagulation pathway, safety problems such as bleeding, allergy and gastrointestinal tract side reactions exist, and fucoidan only acts on an endogenous blood coagulation pathway, so that the risk of bleeding is greatly reduced from the mechanism. In addition, the fucoidan is derived from invertebrates and cannot be degraded by enzymes in mammals, so that oral administration is expected to be realized, and the compliance of patients is greatly improved.

This invention relates to the field of biomedicine, and particularly to a hirudin gene, expression vector, engineered bacteria, recombinant protein, and its applications and production methods, all possessing anticoagulant, thrombolytic, and antioxidant effects. The nucleotide sequence of the hirudin gene is shown in SEQ ID NO: 1. The amino acid sequence of the recombinant protein is shown in SEQ ID NO: 2. The recombinant protein of the hirudin gene provided by this invention, possessing anticoagulant, thrombolytic, and antioxidant effects, exhibits a high concentration of recombinant hirudin, significantly increasing the yield of recombinant hirudin. The recombinant hirudin also demonstrates strong in vitro anticoagulant and thrombolytic activity, indicating broad market prospects.

The subject of the invention is a block copolymer of poly(sodium styrene sulphonate) and poly(acrylic acid) functionalized with a phenylazopyrazole substituent defined by formula (I). The invention also comprises the use of a copolymer for the preparation of a drug for use in controlling the blood clotting process, and its use in the treatment or prevention of diseases requiring control of the blood clotting process.

The present application relates to the technical field of medicine, and provides a potency calibration method of haena polysaccharide.The present application adopts p-nitroaniline colorimetric method to test the haena polysaccharide activity curve, wherein the abscissa of the haena polysaccharide activity curve is the reciprocal of the weight of haena polysaccharide, and the ordinate is the logarithm of the residual activity unit of FVIII; the weight of haena polysaccharide for inhibiting the iFXase (endogenous factor X enzyme) activity of 1 IU FVIII is taken as 1 potency unit, and the potency of haena polysaccharide is obtained according to the haena polysaccharide activity curve.The potency calibration method provided by the present application can accurately calibrate the anticoagulant activity intensity of haena polysaccharide, and has important significance in aspects of guaranteeing drug efficacy, drug safety and quality control.

The invention discloses a recombinant leech polypeptide with high anticoagulant activity as well as a preparation method and application thereof. A PelB-T2Y-N47K-N52Y mutant is constructed by carrying out structural optimization on a hirudin HV2 sequence, a polypeptide product with relatively high anticoagulant activity is obtained after induced expression and nickel affinity chromatography purification are carried out in a prokaryotic expression system, and the anticoagulant activity of the polypeptide product reaches 9135.8 ATU / mg and is remarkably superior to that of an initial sequence HV2. Meanwhile, the polypeptide shows a lower IC50 value in a hydroxyl radical scavenging experiment, and has the activity of inhibiting generation of AGEs in a non-enzymatic saccharification model. Molecular docking and binding free energy analysis show that the combination of the mutant and thrombin (PDB ID: 4HTC) is more stable. The leech polypeptide disclosed by the invention has the potential of being developed into a multifunctional bioactive peptide preparation, and is suitable for the related fields of anticoagulation, antioxidation and anti-saccharification.

The present invention relates to preparation method for the scale up production of a safe bovine heparin composed by a distinctively selected unfractioned bovine heparin polymers with low 6-O-desulfated glucosamine content and a porcine-like anticoagulant activity and protamine neutralization, and methods of its production and application. This safe bovine heparin (SB Heparin) has a comparable structure and function to the porcine heparin, the clinical usage reference, preventing clinical usage impairments as a safe pharmaceutical product, allowing its use as interchangeably drugs.

The present invention relates to binding molecules capable of reversing the anti-coagulant activity of direct oral anticoagulants in-vivo and in-vitro. The present invention, more particularly concerns binding molecules for reversing the factor Xa inhibition activity of a factor Xa inhibitor such as in a patient to whom a factor Xa inhibitor is administered. The present invention is further related to binding molecules capable of reversing the anti-coagulant activity of direct oral anticoagulants, for use in medicine and diagnostics. More particularly, the present invention concerns binding molecules for use in the treatment or prevention of bleeding, and for use in the detection and quantification of such molecules, e.g. in human plasma. The present invention further provides a method for detecting a factor Xa inhibitor in a sample.

The invention belongs to the field of genetic engineering and synthetic biology, and particularly relates to tyrosine sulfation modified high-activity recombinant hirudin, saccharomyces cerevisiae engineering bacteria and application. According to the invention, saccharomyces cerevisiae is taken as a chassis cell, tyrosine protein sulfotransferase and related enzymes and transport systems are introduced through genetic engineering modification, efficient sulfation modification of the hirudin Tyr63 site is realized, and the anticoagulant activity of the hirudin Tyr63 site is remarkably improved. The method overcomes the limitations of high extraction cost of natural hirudin and lack of modification function in the existing recombination technology. In addition, the tyrosine sulfation modified high-activity recombinant hirudin provided by the invention shows an excellent antioxidant effect in the field of beauty and skin care, and experiments prove that the antioxidant activity of the tyrosine sulfation modified high-activity recombinant hirudin is equivalent to that of natural hirudin. The invention provides an innovative solution for low-cost and large-scale production of high-activity hirudin, and has the application potentials of medicines and cosmetics.

The invention discloses a preparation method of Panax stipuleanatus protein, which comprises the following steps: adding 0.01 mol / L NaOH solution with pH of 9 into Panax stipuleanatus powder, uniformly mixing, centrifuging at 4 DEG C, and taking filter residue; adding an ethanol solution with the volume concentration of 70-80% into the filter residues, extracting and centrifuging at 4 DEG C to take the filter residues, adding a NaCl solution with the mass concentration of 2-4% into the filter residues, extracting and centrifuging at 4 DEG C, taking the filter residues again, adding water into the filter residues, extracting and centrifuging at 4 DEG C, and taking supernate; the supernatant is dialyzed for 12-24 h at the temperature of 4 DEG C with a dialysis bag with the molecular weight cut-off of 10 kDa, inner filtrate is collected, freeze drying is conducted, then the panax stipuleanatus protein is prepared, anticoagulant activity detection is conducted on the panax stipuleanatus protein, and the protein has the good anticoagulant effect; the preparation method disclosed by the invention is simple to operate, provides a new way for preparing a novel anticoagulant drug, and has a wide application prospect.

The invention belongs to the field of genetic engineering and synthetic biology, and particularly relates to tyrosine sulfation modified high-activity recombinant hirudin, saccharomyces cerevisiae engineering bacteria and application. According to the invention, saccharomyces cerevisiae is taken as a chassis cell, tyrosine protein sulfotransferase and related enzymes and transport systems are introduced through genetic engineering modification, efficient sulfation modification of the hirudin Tyr63 site is realized, and the anticoagulant activity of the hirudin Tyr63 site is remarkably improved. The method overcomes the limitations of high extraction cost of natural hirudin and lack of modification function in the existing recombination technology. In addition, the tyrosine sulfation modified high-activity recombinant hirudin provided by the invention shows an excellent antioxidant effect in the field of beauty and skin care, and experiments prove that the antioxidant activity of the tyrosine sulfation modified high-activity recombinant hirudin is equivalent to that of natural hirudin. The invention provides an innovative solution for low-cost and large-scale production of high-activity hirudin, and has the application potentials of medicines and cosmetics.

The present invention relates to a monoclonal antibody having an anticoagulant activity and the use thereof. The antibody or a fragment comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region of the antibody or fragment has an amino acid sequence as shown in SEQ ID NO: 1, and the heavy chain variable region has an amino acid sequence as shown in SEQ ID NO: 3. The monoclonal antibody of the present invention has the effect of inhibiting the activity of a common coagulation pathway in the coagulation cascade reaction, and can be used in the prevention and treatment of thrombotic diseases.

The invention discloses an anticoagulant active site of massa medicata fermentata fujianensis as well as a preparation method and application thereof, and belongs to the field of biological medicines. The massa medicata fermentata fujianensis is used for preparing anticoagulant drugs for the first time. The method comprises the following steps: soaking a fermented mass raw material with methanol, performing reflux extraction, and recovering a solvent under reduced pressure to obtain a total extract; and separating the total extract by adopting macroporous adsorption resin column chromatography to obtain a water part, a 30% methanol part, a 50% methanol part and a pure methanol part. Platelet aggregation detection results show that all the parts can inhibit ADP-induced platelet aggregation, and the anticoagulant effect of the 30% methanol part is most prominent and is similar to that of a positive drug. According to the preparation method of the anticoagulant active part of the massa medicata fermentata fujianensis, the anticoagulant active part in the massa medicata fermentata fujianensis can be effectively separated and enriched, and the obtained anticoagulant active part can be applied to preparation of anticoagulant drugs and has good industrial application prospects and clinical value.

The present invention provides antibodies against activated protein C (APC). Certain disclosed antibodies inhibit the anticoagulant activity of APC while preserving its beneficial cytoprotective functions. The present invention also provides nucleic acids, vectors, and host cells for producing the antibodies disclosed herein, as well as methods of using the antibodies to treat medical conditions such as bleeding, sepsis, and inflammation.

A heparin-like ternary copolymer containing zwitterionic, sulfonic, and carboxyl side chains is prepared by atom transfer radical polymerization (ATRP) from a first monomer containing a sulfonic acid group and an olefinic group, a second monomer containing an oxygen-carbon chain-linked zwitterionic group and an olefinic group, and a third monomer containing an oxygen-carbon chain-linked carboxyl side chain and an olefinic group. This ternary copolymer has a heparin-like structure and utilizes zwitterionic groups that mimic the hydrophilicity of cell membrane lecithin and negatively charged sulfonic acid groups with anticoagulant activity to inhibit both intrinsically and exogenously induced coagulation reactions. Anticoagulant coatings prepared from this copolymer can largely address the problem of zwitterionic polymers alone having poor anticoagulant properties and being unable to meet the anticoagulant requirements of biomedical devices. It also provides an effective solution to the multiple side effects and adverse reactions that systemic anticoagulants are prone to.

The invention belongs to the technical field of anticoagulant drugs, and particularly relates to lumbricus protein with anticoagulant activity as well as a preparation method and application of the lumbricus protein. The lumbricus protein provided by the invention contains at least one of a c71616 protein with an amino acid sequence as shown in SEQ ID No. 1, a c128686 protein with an amino acid sequence as shown in SEQ ID No. 2, a c58213 protein with an amino acid sequence as shown in SEQ ID No. 3 and a c118923 protein with an amino acid sequence as shown in SEQ ID No. 4. The protein can effectively inhibit thrombin activity and interfere formation of fibrin so as to act on a key link of a blood coagulation process. The protein can be obtained through recombinant expression and purification, and has high expression efficiency and stability, so that the protein can be used as an active component to be applied to development of anticoagulant drugs, surface anticoagulant coatings and other products.

The present invention discloses an anticoagulant peptide, its preparation method and application. The present invention optimizes the extraction and separation method of anticoagulant activity of Eupolyphaga sinensis, thereby detecting ultra-trace anticoagulant peptide RF7 from complex crude peptide extracts, and using the NanoLC-Q-Orbitrap-MS / MS method to perform high-throughput analysis and identification of the peptide sequences of the screened active anticoagulant peptides. Through the search and comparison of the intelligent data processing system and the polypeptide database, the biological information and pharmaceutical components of the anticoagulant peptide are comprehensively analyzed. Finally, by constructing molecular level, cell and animal models, pharmacokinetic and pharmacodynamic studies are carried out on the anticoagulant peptides with obvious activity to evaluate the safety and effectiveness of the drug, laying a research foundation for the development and drug-likeness evaluation of new anticoagulant drugs.

The application discloses a salt-free production technology of heparin sodium crude product extracted by enzyme hydrolysis method of biological enzyme fermentation, and belongs to the technical field of production of heparin sodium crude product extracted from pig small intestinal mucosa. The application adopts controllable biological fermentation technology of bioactive enzyme combined with enzyme hydrolysis method to extract heparin sodium biological macromolecule with anticoagulant biological activity. The yield of anticoagulant activity in the obtained enzyme hydrolysis liquid is 96,000 USPU / root, and the yield of dried solid heparin sodium crude product is 82,000 US standard units / root. The technical scheme of the application greatly improves the yield of heparin sodium crude product, simultaneously reduces 73% of wastewater and 99% of chloride ion content, greatly reduces the difficulty of wastewater treatment, makes it possible to change waste into treasure by comprehensive recycling of wastewater, and is favorable for industrial application.

The preparation method comprises the following specific steps: S1, weighing L-borneol crystal substances, D-borneol crystal substances and Baijiu, and mixing the L-borneol crystal substances, the D-borneol crystal substances and the Baijiu; s2, putting the mixture into a pressure cooker filled with distilled water, heating the interior of the pressure cooker to a boiling state of 100-105 DEG C by using big fire, and continuously cooking for 4-8 hours by using small fire; and S3, carrying out cooling crystallization treatment to obtain the racemic borneol.

The invention discloses tanshinone derivatives with antithrombotic activity as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The tanshinone derivative is obtained by structurally modifying tanshinone IIA and cryptotanshinone, the polarity and pharmacological activity are remarkably improved, the bioavailability is greatly improved, the problem that the tanshinone IIA and the cryptotanshinone are limited in clinical application is solved, and the problems that the tanshinone IIA and the cryptotanshinone are high in lipid solubility, difficult to prepare into proper dosage forms, affected in absorption and distribution of drugs, short in half-life period, required to be frequently administered and poor in bioavailability are solved. And inconvenience is brought to the patient. Part of tanshinone derivatives have remarkable anticoagulant activity; the synthetic method is simple to operate and controllable in condition, a series of derivatives can be efficiently synthesized, and the derivatives have the biological activities of anticoagulation, platelet aggregation resistance, thrombosis resistance and the like, are the key for the antithrombotic drugs to play a therapeutic role, provide efficient and safe candidate drugs for cardiovascular and cerebrovascular diseases, and have remarkable technical advantages and huge market potential.

The present invention provides antibodies against activated protein C (APC). Certain disclosed antibodies inhibit the anticoagulant activity of APC while maintaining its beneficial cytoprotective functions. The present invention also provides nucleic acids, vectors, and host cells for producing the antibodies disclosed herein, as well as methods of using the antibodies to treat medical conditions such as hemorrhage, sepsis, and inflammation.

The invention discloses a high-anticoagulation active peptide and a preparation method and application thereof, and relates to a shrimp paste high-anticoagulation active protein component based on compound enzyme assisted rapid fermentation and a preparation method thereof, and a preparation method of a high-anticoagulation active shrimp paste. The method for rapidly preparing the shrimp paste with high anticoagulation activity and assistance of enzymolysis is obtained by screening enzymolysis-assisted fermentation conditions such as shrimp producing areas, enzyme types, enzymolysis temperature, enzymolysis time, enzymolysis pH and the like. After purification is carried out by combining ultrafiltration, gel chromatography and non-denaturing electrophoresis, the anticoagulant active peptides PSEPSKPVTCKPR and GIAEGCDYPWR are identified by combining proteomics and a virtual screening technology. According to the invention, not only is the preparation method of the high-anticoagulant-activity enzymatic-method-assisted rapid fermentation shrimp paste obtained, but also two novel anticoagulant peptides are innovatively separated from the shrimp paste, and a novel biological agent development basis is provided for treatment of thrombotic diseases.

The present invention provides antibodies against activated protein C (APC). Certain disclosed antibodies inhibit the anticoagulant activity of APC while preserving its beneficial cytoprotective functions. The present invention also provides nucleic acids, vectors, and host cells for producing the antibodies disclosed herein, as well as methods of using the antibodies to treat medical conditions such as bleeding, sepsis, and inflammation.

The invention relates to the technical field of microorganisms, and particularly discloses a marine bacterium Bacillus sp.4SP1 as well as a plasmin production method and application thereof, the strain is preserved in China General Microbiological Culture Collection Center on November 6, 2024, the preservation number is CGMCC No.32524, the preservation unit address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing, the preservation number is CGMCC No.32524, and the preservation number is CGMCC No.32524. Institute of Microbiology, Chinese Academy of Sciences; the bacterial colony of the bacterium is white and oval, has a rough surface, a convex center and irregular edges, and is gram-positive bacterium; the optimal growth conditions of the strain 4SP1 are as follows: the yeast is 5g / L, the tryptone is 10g / L, the pH value is 7.0, and the culture temperature is 40 DEG C. The plasmin produced by the marine bacterium Bacillus sp.4SP1 disclosed by the invention has thrombolysis and anticoagulant activity, and has a good application prospect.