36 results about "Recombinant Hirudin" patented technology

The invention, belonging to the field of cosmetics, relates to a new use of leech, specifically to the use of leech as main raw material for cleaning skin, a skin cleaning composition prepared based on the characteristic and a preparation method. According to the invention, microcirculation of skin is improved by means of the blood circulation-promoting and stasis-dissolving effect of leech extract, natural hirudin and recombinant hirudin in order to relax skin as well as thoroughly cleanse dirt, blackheads and whiteheads in pores and aged cuticles (commonly referred to as dead skin) on the surface of skin and the like, therefore, superior foundation is laid for basic skincare. Based upon the above new use, the leech extract, the natural hirudin and the recombinant hirudin can be prepared to cosmetic cleansing products, including face cleansing cream, face cleansing gel, scrubbing cream, cream for removing dead skin, body wash, makeup remover, perfumed soap, etc.

The invention discloses a recombinant bacterium for producing sulfo-recombinant hirudin as well as a preparation method and application of the recombinant bacterium, and belongs to the technical field of genetic engineering. In order to improve the synthesis yield of the sulfated recombinant hirudin and reduce the production cost, the invention provides the recombinant bacterium for producing the sulfo-recombinant hirudin, wherein the recombinant bacterium overexpresses a sulfotransferase gene and a hirudin gene, and an original strain is escherichia coli; and the nucleotide sequence of the hirudin gene is one of SEQ ID No.1 to SEQ ID No.3, and the nucleotide sequence of the sulfotransferase gene is as shown in SEQ ID No.4. According to the method, the activity of the recombinant hirudin is remarkably improved, a one-step purification method of the hirudin is realized, a recombinant hirudin product with relatively high purity can be obtained at one time, and the pressure of subsequent purification is greatly reduced.

The invention relates to the field of preparation, in particular to a leech colon targeted oral preparation prepared by taking leech extract, natural hirudin and recombinant hirudin as active ingredients and a preparation method thereof. The invention aims at solving the technical problems that a leech colon targeted oral preparation which is released and absorbed and gives curative effect to be played at a colon part is provided and bioavailability of oral administration is improved. The preparation method comprises the following steps of: preparing the active ingredients into a coating front midbody, then applying a colon targeted coating system onto the coating front midbody, wherein the coating system comprises an inner coating layer and an outer coating layer; the inner coating layer is mixture of 1 part of guar gum, 1-3 parts of sodium alginate and 1-3 parts of inulin, the mixture is dissolved with water and is prepared into coating solution with the concentration of 2-5%, and weight of the coating is increased by 3-9%; and the outer coating layer is mixture of a film forming agent and a porogenic agent, the outer coating layer is in a weight proportion relation that 6-8 parts of the film forming agent and 2-4 parts of the porogenic agent are adopted, and the weight of the coating is increased by 3-15%. The oral preparation provided by the invention can be released and absorbed at the colon part, and the bioavailability can be improved.

The invention discloses a repairing eye cream. The repairing eye cream contains the following raw materials in parts by weight: 35-40 parts of cyclopentasiloxane, 24-28 parts of recombinant hirudin, 15-20 parts of a repairing agent, 1-3 parts of hydroxydecyl ubiquinone, 8-10 parts of an aloe extracting solution, 5-7 parts of butanediol and 30-40 parts of water. The repairing eye cream has excellent preservation stability and good repairing effect and is capable of effectively moistening eye skin, fading eye microgrooves and black eyes and eliminating fat granules.

The invention discloses a leech tyrosine sulphonate transferase gene and application thereof, and belongs to the field of biomedicine. The leech tyrosine sulphonate transferase gene is characterized in that on the basis of a hirudo nipponia transcriptome database, a leech tyrosine sulphonate transferase cDNA sequence is excavated and identified through a bioinformatics means, and on this basis, the leech tyrosine sulfotransferase is synthesized through codon optimization, gene synthesis and microbial cell expression, and the recombinant hirudin is catalyzed by the leech tyrosine sulfotransferase to be subjected to sulfonation modification to obtain the natural hirudin. The catalytic ability of the transferase is greatly improved compared with that of acyl sulfonate transferase, human-derived tyrosine sulfonate transferase and bovine-derived tyrosine sulfonate transferase. Compared with other tyrosine sulfonic acid transferase, the leech tyrosine sulfonic acid transferase obtained in the invention has stronger substrate preference for recombinant hirudin, is suitable for large-scale preparation of natural hirudin, and provides a new way for synthesis of natural hirudin.

The invention discloses a recombinant hirudin fusion protein with targeting and long-acting functions and a coding gene and application thereof. The fusion protein is formed by fusing a 'procoagulant platelet' targeting binding peptide, hirudin and an albumin binding domain. The fusion protein has the property of a prodrug, can be quickly activated in vivo and exerts an antithrombotic effect; the effect of the fusion protein is equivalent to that of the hirudin, but the duration of the anticoagulant effect is significantly prolonged compared with that of the hirudin, the bleeding time is significantly reduced compared with that of the hirudin and is close to the level of normal saline, the systemic bleeding risk is very low, and the fusion protein has a broad application prospect.

The invention discloses a recombinant hirudin eye drop. The recombinant hirudin eye drop takes recombinant hirudin as an effective component and sodium hyaluronate as a thickening agent, and also contains an isoosmotic adjusting agent, a metal complexing agent, a preservative and a pharmaceutically acceptable carrier; and the mass ratio of the sodium hyaluronate to the recombinant hirudin to the isoosmotic adjusting agent to the metal complexing agent to the preservative in the eye drop is 10 to 2 to 15.27 to 50 to 3. The preparation method comprises the following steps: dissolving the sodium hyaluronate, the isoosmotic adjusting agent, the metal complexing agent and the preservative into the pharmaceutically acceptable carrier to prepare a blank matrix of the eye drop, dissolving the recombinant hirudin into the blank matrix of the eye drop, mixing evenly, filtering, degerming and sub-packaging to prepare the recombinant hirudin eye drop. According to the eye drop disclosed by the invention, on one hand, the recombinant hirudin can be retainedin a focal zone for a long period of time; on the other hand, irritation is not generated to sensitive eye tissues; and meanwhile, the storage stability of the recombinant hirudin eye drop can also be ensured.

The invention belongs to the field of biotechnology and particularly relates to a low-immunogenicity recombinant hirudin mutant and a preparation method thereof. The method comprises the following steps of: identifying hirudin dominant antigen epitopes by a phage peptide library and a Pepscan (SPOTS) technology; scanning critical combining residues of dominant antigen epitopes by alanine (ALA) canning technology and then positioning accurately; and removing immunogenicity by using a site-specific mutagenesis technology so as to obtain hirudin mutant, wherein according to obtained hirudin mutant of which the anticoagulant rate activity is not changed basically but the immunogenicity and the immunogenicity are obviously reduced. The low-immunogenicity recombinant hirudin mutant and a preparation method thereof have important signification for overcoming the immunogenicity problem of the conventional hirudin medicaments, improving the clinical treatment effect of hirudin medicaments, and fully exerting clinical using values of the hirudin medicaments.

The invention relates to the technical field of separation and purification of biological medicines, in particular to a method for removing endotoxin in a recombinant hirudin protein solution. The method performs separation by adopting anion exchange chromatography and comprises the following steps: injection: the recombinant hirudin protein solution is taken for injection, the injection volume is 25%-40% of the maximum load of a chromatographic column, and washing is performed with an equilibrium solution to perform column equilibrium; elution: isocratic elution is performed with an eluent, and an outflow part is collected to obtain a recombinant hirudin eluent; and column regeneration: isocratic elution is performed with regenerated liquid, and an outflow part is collected to obtain an endotoxin eluent. According to the method, according to the strength of adsorption of recombinant hirudin and endotoxin with an anion exchanger under neutral conditions, the isocratic elution is performed by increasing salt concentration in a mobile phase, anda good removal effect is achieved by adjusting the injection volume, thus guaranteeing a relatively high protein recovery rate, and meanwhile, regeneration of the chromatographic column is implemented. The method is simple, efficient, low in cost and suitable for industrial production.

The invention belongs to the technical field of biological medicines, and relates to a recombinant hirudin microneedle drug delivery system and a preparation method thereof. The recombinant hirudin microneedle drug delivery system comprises a backing patch and a microneedle body located at the bottom of the backing patch, wherein recombinant hirudin is packaged in the microneedle body, and a microneedle matrix is a water-soluble biodegradable high polymer material. Compared with an existing drug delivery mode, the microneedle drug delivery system has the advantages that pain caused by repeatedinjection can be remarkably relieved during use, and the drug is stable in activity and convenient to store and transport.

The invention discloses tumor-specific recombinant hirudin, of which an N-terminal contains an oligopeptide sequence PLGI, which can be recognized and cleaved by matrix metalloproteinase 9 (MMP9), wherein hirudin used for recombination may be a hirudin isomer, a hirudin mutant, truncated hirudin peptide, gene-modified hirudin or hirudin fusion protein. Correspondingly, the invention provides a preparation method of the tumor-specific recombinant hirudin and applications thereof in preparation of antitumor medicines, and also provides a preparation method and an application of adoptive immune-cells for expression of the tumor-specific recombinant hirudin. The recombinant hirudin herein allows anticoagulation activity of the hirudin is released in a tumor targeted manner, thus forming a microenvironment which is not liable to form thrombus, and even dissolving formed small thrombus to achieve the functions of anticoagulation, thrombolysis and prevention of tumor cell metastasis, thereby overcoming the risk of systematic hemorrhage due to systemic administration of wild-type hirudin.

The invention provides an application of hirudin for repairing skin red blood filament or hormone face, a composition and a preparation method thereof, natural hirudin or recombinant hirudin are takenas main active ingredients, and one or two of rhubarb extract and allantoin are taken as active ingredients together for repairing skin red blood filament or hormone face problems. The invention hasthe beneficial effects that hirudin is the main active ingredient, emodin and allantoin are combined to neutralize inflammatory factors, inhibit the secretion and diffusion of inflammatory factors, rebuild skin barrier, and improve the problems of red blood filament and hormonal face.