37 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.

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 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 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 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.

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.

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 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.

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.

The invention provides an anticoagulant polypeptide HS9 and application thereof, and belongs to the technical field of functional polypeptides. The amino acid sequence of the anticoagulant polypeptide HS9 is as shown in SEQ ID NO: 1. The anticoagulant polypeptide HS9 has strong anticoagulant activity, has the efficacy of inhibiting thrombosis in gestation period and reducing bleeding risk, and also has the characteristics of simple structure and convenience in preparation. Therefore, the invention provides application of the anticoagulant polypeptide HS9 in preparation of antithrombotic drugs or drugs for preventing and / or treating blood coagulation dysfunction diseases.

The present application relates to the field of living extraction technology, in particular to a leech living body collection and anticoagulant activity substance synchronous extraction process, which comprises the following steps: a large number of living leeches are poured into a shunt box and are shunted by a shunt channel and then fall into a row of a plurality of object cavities; a motor drives a transfer cylinder to rotate intermittently to pick up the living leeches, and the transfer cylinder adsorbs the leeches; when the leeches fall into a plurality of collection cavities of a collection box, the leeches are attracted by a blood-containing artificial membrane and bite, and the saliva is secreted by the current stimulation of an electrode sheet; the saliva flows into a liquid collecting box and is filtered by a filter membrane in the liquid collecting box, the filtered saliva is directly introduced into a low-temperature container through a bottom port connecting pipeline of the liquid collecting box for collection, and the leech anticoagulant is obtained by centrifugal impurity removal. The funnel structure of the shunt box is combined with the flow guide sleeve to increase the single processing capacity, the air pressure adsorption and the intermittent rotation mechanism of the transfer cylinder are used to complete individual transfer, and the electrode sheet and the artificial membrane integrated stimulation system are used to achieve directional saliva collection.

The application discloses a fish swim bladder chondroitin sulfate-Dermatan sulfate hybrid chain and a preparation method and application thereof, and relates to the technical field of medicines. The fish swim bladder chondroitin sulfate (CS) / dermatan sulfate (DS) hybrid chain with anticoagulant activity is composed of iduronic acid, N-acetylgalactosamine, glucuronic acid and N-acetylglucosamine, and the CS / DS hybrid chain is derived from a fish swim bladder, is easily soluble in water, has strong anticoagulant activity, and can be used for preparing medicines for preventing and / or treating thrombotic cardiovascular diseases.

This invention discloses a high-yield, dual-effect earthworm protein small molecule peptide and its preparation method, specifically relating to the field of biomedical technology. The preparation method includes: S1, synergistic enzymatic hydrolysis: earthworm protein undergoes autolysis, followed by enzymatic hydrolysis with a complex enzyme of flavor protease and alkaline protease; S2, charge-size bi-stage membrane separation: the hydrolysate is sequentially passed through a 5kDa first-stage ultrafiltration membrane and a 1kDa second-stage ultrafiltration membrane modified with a positive charge; S3, dual-activity-guided purification: using anticoagulant activity and dipeptidyl peptidase-IV inhibitory activity as tracking indicators, the highly active components are collected. The small molecule peptide obtained by this invention has a molecular weight of 400-800 Da, contains novel sequences shown in SEQ ID NO:1 and SEQ ID NO:2, and simultaneously possesses anticoagulant activity and DPP-IV inhibitory activity, with a yield of 17.2-19.1%, which is more than 30% higher than conventional methods. The process of this invention is stable and has good prospects for industrialization.

The present invention relates to supramolecular agents presenting on demand reversibility of activity and related methods. The invention in particular relates to supramolecular agents with anticoagulant activity which can be reversed with the use of an antidote agent.

A leech extract is a salivary gland extract of poecilobdella manillensis, and is obtained through living body cleaning, gland taking and smashing, purification, separation and multi-step purification. The invention further discloses biological polypeptide composite herbal extract shampoo containing the leech extract and a preparation method of the biological polypeptide composite herbal extract shampoo. The anticoagulant activity of the leech extract is about 1500U, the anticoagulant thrombolytic efficacy is doubled compared with that of a common leech extract, the blood coagulation time can be prolonged, thrombosis can be inhibited, and the fibrinolytic activity can be improved; the effects of improving scalp microcirculation, preventing fibrinogen from forming thrombus, dredging blood capillaries, increasing scalp blood flow, promoting hair follicle nutrition supply and reducing alopecia are achieved.

An anticoagulant phenyl-linked heparin oligosaccharide dimer, a preparation method therefor and use thereof are provided. The anticoagulant phenyl-linked heparin oligosaccharide dimer has a structure as represented by general formula I, and the phenyl may be replaced with substituted phenyl, heteroaryl or substituted heteroaryl. The described phenyl-linked heparin oligosaccharide dimer may be synthesized in fewer steps, has a significantly higher total yield, potent and specific antithrombin-dependent anti-Factor Xa activity without significant anti-FactorIIa activity, the anticoagulant activity may be efficiently neutralized by protamine sulfate, has excellent pharmacokinetic profiles, making it suitable for the preparation of novel anticoagulant and antithrombotic drugs that are cost-effective, higher-quality and safer.

This invention discloses a method for preparing alginate sulfate oligomers. By strictly controlling the addition amounts of TEMPO, sodium bromide, and sodium hypochlorite under specific conditions, highly efficient degradation of alginate sulfate is achieved. Specifically, the mass ratio of TEMPO to alginate sulfate is 0.5-5%, the mass ratio of sodium bromide to alginate sulfate is 5-40%, and the mass ratio of alginate sulfate to sodium hypochlorite is 1 g:0.1-10 mmol / L. No strong acid or enzyme preparations are required, and no microwave or ultrasonic auxiliary methods are relied upon. Oligomers with an average molecular weight below 80 kDa can be obtained within 1 hour, and oligomers with an average molecular weight of approximately 20 kDa can be obtained within 4 hours, while maintaining a sulfate content of approximately 25%. The method causes almost no damage to the functional groups of alginate polysaccharides, and the prepared oligomers exhibit significant effects in in vitro anticoagulant activity.