50 results about "Anticoagulant activity" patented technology

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 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.

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 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.

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 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.

This application discloses a complex polysaccharide, its preparation method, and its applications. It discloses both a novel use of sulfated kudzu polysaccharide as a raw material for preparing drugs with anticoagulant effects and a complex polysaccharide comprising sulfated polysaccharide and polysaccharide fragments; wherein the sulfated polysaccharide includes sulfated kudzu polysaccharide and / or sulfated Bletilla striata polysaccharide; and the polysaccharide fragments include glycosaminoglycan fragments. This application modifies the structure of natural polysaccharides to improve their structural properties, preparing and screening complex polysaccharides with anticoagulant activity. The components in the complex polysaccharide work synergistically, significantly enhancing the anticoagulant effect and broadening the application fields of polysaccharides, primarily for the development of anticoagulant drugs and anticoagulant medical devices.

The present invention discloses a bio-fermentation and enzymatic hydrolysis production technology for extracting crude sodium heparin from the mucosa of the pig small intestine, belonging to the technical field of production for extracting crude sodium heparin. The present invention uses a bio-active enzyme controlled bio-fermentation technology combined with an enzymatic hydrolysis method to extract heparin sodium biomacromolecules with anticoagulant biological activity. The anticoagulant activity yield in the resulting enzymatic hydrolyzate is 114,000 USPU / root, and the yield of the dried solid crude sodium heparin obtained is 88,843 USPU / root, or 1,126 roots / 100 million units. Compared with the process of enzymatic hydrolysis alone without the combined bio-active enzyme bio-fermentation technology, the yield is increased by 22.2%. This demonstrates that the bio-active enzyme controlled bio-fermentation technology combined with the enzymatic hydrolysis extraction technology has significant advantages in industrial production. At the same time, the salt usage in production is 1.2%, which is a 66% decrease compared to the common salt usage in existing industry methods, reducing the difficulty of wastewater treatment and facilitating industrial application.

This invention discloses an earthworm protein peptide with in vitro thrombin-inhibiting activity and its preparation method, belonging to the field of functional food technology. The method first pre-treats earthworm raw materials to obtain earthworm protein; then, it uses pepsin and alkaline protease for stepwise enzymatic hydrolysis under varying temperature and pH, and ultrafiltration to collect short peptide solutions with a molecular weight below 3 kDa; the short peptide solution is specifically captured using an affinity chromatography material coupled with bovine thrombin, followed by elution, nanofiltration desalting concentration, and spray drying to obtain the final product. This invention fully releases the peptides through a dual enzymatic hydrolysis process and utilizes thrombin affinity chromatography to directionally enrich peptides with specific inhibitory activity, effectively solving the problems of low content and poor targeting of active ingredients in earthworm protein peptides in existing technologies, and significantly improving the purity and in vitro anticoagulant activity of the product.

The present invention belongs to the field of genetic engineering and synthetic biology, and specifically relates to a tyrosine sulfated highly active recombinant hirudin, brewer's yeast engineered bacteria and applications. The present invention uses brewer's yeast as a chassis cell, and through genetic engineering modification, introduces tyrosine protein sulfonate transferase and related enzymes and transport systems to achieve efficient sulfate modification of the Tyr63 site of hirudin, significantly improving its anticoagulant activity. This method overcomes the limitations of the high cost of extracting natural hirudin and the lack of modification function of existing recombinant technologies. In addition, the tyrosine sulfated highly active recombinant hirudin provided by the present invention exhibits excellent antioxidant efficacy in the field of beauty and skin care, and experiments have shown that its antioxidant activity is equivalent to that of natural hirudin. The present invention provides an innovative solution for the low-cost, large-scale production of highly active hirudin, and has potential for application in both medicine and cosmetics.

The invention discloses a whey protein peptide with high anticoagulant activity and a blood fat reducing effect and a preparation method thereof. The preparation method comprises the following steps: firstly carrying out pulsed electric field pretreatment on a whey protein aqueous solution, then hydrolyzing the pretreated whey protein aqueous solution by using bromelain, and freeze-drying to obtain the whey protein peptide. According to the method, bromelain is selected to carry out enzymolysis on the whey protein aqueous solution, and the whey protein aqueous solution is pretreated by using the pulsed electric field before enzymolysis, so that the use of the pulsed electric field is beneficial to promoting the exposure of peptide chain segments with anticoagulation and blood fat reduction functions, and the anticoagulation and blood fat reduction capabilities of the whey protein peptide are remarkably improved. The method breaks through the limitation of the traditional process, and overcomes the problems of low efficiency and limited active peptide release of a single enzymolysis method through the synergistic effect of the pulsed electric field pretreatment and the enzymolysis technology. The non-thermal effect of the pulsed electric field can directionally change the space structure of the whey protein, more enzyme cutting sites are exposed, and meanwhile protein denaturation caused by high-temperature treatment is avoided.

The present disclosure provides compositions and methods related to nucleic acid molecules having therapeutic aptamers. In particular, the present disclosure provides nucleic acids molecules comprising one or more aptamers having anticoagulant activity, as well as corresponding nucleic acid antidotes, for the modulation of blood coagulation in the context of disease and surgical intervention.

The invention belongs to the technical field of biological medicine, and discloses application of an active substance in preparation of a VKOR inhibitor and / or a vitamin K antagonist and an anticoagulant drug. The invention provides application of an active substance shown in a formula (1) or pharmaceutically acceptable salt thereof in preparation of a vitamin K epoxide reductase inhibitor. The active substance has VKOR inhibitory activity, can prevent liver from synthesizing blood coagulation factors and activating anticoagulant factors by blocking reduction of oxidized vitamin K and inhibiting circulation of vitamin K, has potential anticoagulant activity, and has the advantages of mild anticoagulant effect, low probability of occurrence of adverse hemorrhage events, good anticoagulant effect, high safety and the like. The compound has a good application prospect in preparation of anticoagulants and / or medicines for treating thromboembolic diseases.

The invention provides an anticoagulant polypeptide OM20 and application thereof, and belongs to the technical field of biological medicine. The invention provides a polypeptide OM20 with anticoagulant activity, the polypeptide takes SEQ ID No.1 as a basic sequence, and modification, substitution, addition or deletion and modification can be carried out on the basis of the basic sequence. The polypeptide molecule disclosed by the invention can be used for remarkably inhibiting plasma recalcification coagulation time and remarkably delaying APTT time, and has a function of preventing thrombosis. The OM20 targets thrombin, and has the advantages of no bleeding risk, no cytotoxicity and high safety. In conclusion, the OM20 has anticoagulant activity, can be developed into a potential anticoagulant drug molecule as a resource, and is applied to prevention and treatment of thrombosis of human beings and other animals. Secondly, the OM20 acts on the thrombin in a targeting manner, and can be applied to research on thrombotic diseases caused by the thrombin and used as a potential therapeutic drug.

This invention provides a sheep milk casein anticoagulant peptide, its preparation method, and its applications. The sheep milk casein inhibitory peptide is VKETMVPK, VLPVPQK, or EMPFPK. This invention addresses the problems of low safety and high cost of existing clinical anticoagulant drugs, and the low utilization rate of high-value sheep milk casein resources. It fills the technological gap in the preparation of sheep milk casein anticoagulant peptides. The provided sheep milk casein inhibitory peptide has high safety, avoiding the drawbacks of existing drug treatments; efficiently taps the value of sheep milk resources, reducing resource waste; fills a technological gap, establishing a systematic preparation system; has clear and efficient anticoagulant activity with a comprehensive mechanism of action; and has wide applications and great market potential.