53 results about "Urokinase" patented technology

The invention provides a separation and purification method of urokinase, and relates to the technical field of separation and extraction. The separation and purification method comprises the following steps: S1, regulating the pH value of male urine to 8.5, standing, and taking supernate; s2, mixing the supernate with an adsorption material, and eluting with an eluent to obtain a urokinase crude product solution; the adsorption material is a core-shell silica gel polymer, Fe3O4atSiO2-NH2 is taken as a core, and polystyrene is taken as a shell; the eluent is a sodium citrate buffer solution with the concentration of 0.5 to 0.15 M, acetonitrile with the concentration of 1 to 5 weight percent and EDTA (Ethylene Diamine Tetraacetic Acid) with the concentration of 0.5 to 1.5 mM; and S3, carrying out anion exchange column chromatography on the urokinase crude product solution, eluting with an eluent, and precipitating to obtain the urokinase. The urokinase prepared by the method has the advantages of high polymer urokinase content, high recovery rate and high activity yield.

Methods and compositions for treating acute kidney injury in a subject are provided. The methods include measuring or having measured a level of soluble urokinase plasminogen activator receptor (suPAR) in a biological sample from the subject, determining or having determined the level of suPAR in the sample compared to a control suPAR level, and administering a therapeutically effective amount of an agent that antagonizes soluble urokinase plasminogen activator receptor (suPAR) to the subject having an elevated level of suPAR relative to the control suPAR level.

This invention belongs to the field of urea preparation technology and discloses a large-particle urea, composed of the following raw materials in a specific ratio: a 50% concentration of dilute urine solvent, a urokinase activity inhibitor, and a pigment, in a ratio of 100:6:5. This solution, through the careful design of the 50% concentration of dilute urine solvent, the urokinase activity inhibitor, and the pigment, enhances the soil dissolution rate and utilization rate. The addition of the urokinase activity inhibitor effectively inhibits the rapid hydrolysis of urea, reduces ammonia nitrogen loss, improves nitrogen utilization, mitigates negative environmental impacts, and contributes to soil ecological balance. The use of large-particle morphology and strict moisture control technology ensures the product's moisture resistance and storage stability, extending its shelf life. During production, the temperature is precisely controlled at 30℃, which avoids urine crystallization, ensures the full dissolution of the urokinase activity inhibitor, avoids odors generated at high temperatures, reduces environmental pollution, and improves the product's environmental friendliness.

Methods and compositions for treating acute kidney injury in a subject are provided. The methods include measuring or having measured a level of soluble urokinase plasminogen activator receptor (suPAR) in a biological sample from the subject, determining or having determined the level of suPAR in the sample compared to a control suPAR level, and administering a therapeutically effective amount of an agent that antagonizes soluble urokinase plasminogen activator receptor (suPAR) to the subject having an elevated level of suPAR relative to the control suPAR level.

This invention discloses a fusion protein targeting the urokinase-type plasminogen activator receptor and its application in the field of anti-aging. The fusion protein is composed of a hepatocyte growth factor active domain and a humanized anti-uPAR single-domain antibody linked by a flexible linker peptide, and its specific amino acid sequence is SEQ ID NO:3. This fusion protein can specifically bind to uPAR, which is highly expressed on the surface of senescent cells, with intramolar affinity, achieving targeted delivery of the hepatocyte growth factor active domain. In vitro experiments have demonstrated that this protein can significantly reverse the aging phenotype of human fibroblasts, effectively reduce aging-related β-galactosidase activity, p16INK4a protein expression, and interleukin-6 secretion, and promote cell proliferation. In a rapidly aging mouse model, this protein can systematically improve age-related physiological functional decline. The fusion protein of this invention achieves specific targeting and efficient treatment of senescent cells, providing a new solution for the development of anti-aging drugs.

Provided are antibodies that specifically bind to human urokinase-type plasminogen activator receptor (uPAR). In some cases, the antibodies are cross-reactive with one or more non-human animal uPAR polypeptides, such as non-human primate uPAR, e.g., cynomolgus monkey uPAR. Fusion proteins and conjugates comprising the antibodies of the present disclosure are also provided. Methods of using the antibodies, fusion proteins, and conjugates of the present disclosure to treat conditions associated with uPAR expression and / or activity are also provided. In some embodiments, the condition associated with uPAR expression and / or activity is cancer. Non-limiting examples of such cancers include cancers characterized by cancer cells expressing uPAR on their surface, cancers characterized by stromal cells within the tumor microenvironment expressing uPAR on their surface, and the like.

Disclosed herein, are peptides that bind urokinase-type plasminogen activator receptors. The peptides may comprise amino acid sequences of IPPWEAPK (SEQ ID NO: 1), DLAQCQTPTQAAPPTPVSPR (SEQ ID NO: 2), or LHVPLMPAQPAPPK (SEQ ID NO: 3), or retro-inverso amino acid sequences of the above enumerated sequences. Also described herein, are methods of administering compounds comprising peptides that bind urokinase-type plasminogen activator receptors to subjects for the treatment of ovarian cancer and improving wound closure.

This invention relates to the field of biotechnology, specifically to a TruC-T cell that secretes and expresses IL-7 and PF4 to enhance anti-tumor activity and targets uPAR, and its application. Interleukin-7 (IL-7) can effectively promote the development, survival, and proliferation of T cells, while platelet factor 4 (PF4 / CXCL4) can regulate the immune response and chemotactically attract leukocytes to specific sites; the synergistic effect of the two can enhance the anti-tumor function and persistence of T cells. Meanwhile, urokinase-type plasminogen activator receptor (uPAR) is highly expressed in various solid tumors and hematological malignancies, making it an ideal target for tumor therapy; TruC-T cells targeting uPAR can precisely act on tumor tissue, and combined with the immune-enhancing effects of IL-7 and PF4, can significantly improve the efficacy of anti-tumor immune responses, prolong the duration of therapeutic effects, and reduce tumor recurrence.

The current invention relates to a polypeptide comprising an immunoglobulin single variable domain and an urokinase-type plasminogen activator (uPA) domain wherein the immunoglobulin single variable domain is able to bind to fibrin via a fibrin-binding domain.

The application discloses an engineered exosome targeting human urokinase-type plasminogen activator receptor uPAR and a preparation method and application thereof. Firstly, a high-affinity anti-human uPAR single-domain antibody VHH is prepared; then, a human umbilical cord mesenchymal stem cell engineering cell line expressing a VHH-PTGFRN fusion protein is constructed, and the engineered exosome is prepared and purified by combining three-dimensional hypoxic culture, chemical induction and size-exclusion chromatography technology. The engineered exosome comprises four comparative samples, wherein the core sample Exo^(VHH / miR) can simultaneously display the anti-uPAR VHH on the membrane surface to realize targeted delivery and enrich miR-193b-3p in the chamber to realize gene silencing, and the synergistic effect of the two can efficiently remove the uPAR high-expression senescent cells and inhibit the senescence-associated secretory phenotype SASP. The engineered exosome has the advantages of standardized preparation process, high yield and high purity, strong targeting, excellent curative effect and good safety, and can be used for preparing a treatment preparation for anti-aging and aging-related diseases, and has important clinical application value.

The invention provides a preparation method of an aprotinin affinity chromatographic column. Specifically, the aprotinin is used as a raw material and is coupled with CNBr activated agarose gel Focurose 4FF to prepare the aprotinin affinity chromatographic column. The coupling reaction in the preparation method is simple and easy to control, the reaction condition is mild, the operation is convenient, the repeatability is high, and industrial amplification is easy. The affinity column prepared by the invention can completely remove human urinary kallidinogenase in urokinase, and can be repeatedly used for multiple times.

The invention discloses preparation and application of macrophage targeting nanoparticles entrapped with a cell burial function accelerant. The medicine for enhancing the intercellular function of the macrophages is a liver X receptor stimulant, and the liver X receptor stimulant comprises at least one of T0901317, GW3965, Ouabagenin and LXR-623. The surface of the targeting nanoparticle is coated with a macrophage cell membrane, and meanwhile, a urokinase type plasminogen activator receptor (uPAR) targeting ligand is modified. The nanoparticle can be efficiently accumulated at an inflammatory focus part by utilizing a macrophage cell membrane, and the modified targeting ligand is used for targeting the macrophage with damaged cell burial function, so that the cell burial function of the macrophage is enhanced, the polarization of the macrophage to an anti-inflammatory phenotype is promoted, the release of anti-inflammatory and proinflammatory cytokines is regulated, and the anti-inflammatory effect of the nanoparticle is improved. And finally, the inflammatory microenvironment of a focus part is regulated and controlled, so that a good effect of treating autoimmune diseases is achieved.

The utility model discloses a urokinase eluent recovery device which comprises a base, a mounting rack is arranged above the base, an elution tank is fixedly arranged on the base, four groups of symmetrically distributed liquid collecting tanks are arranged on the mounting rack, the liquid collecting tanks and the elution tank are correspondingly distributed, a mounting ring is arranged above the elution tank, and the mounting ring is arranged on the mounting rack. A mounting ring is arranged below the elution tank, a scraping rod is fixedly mounted on the mounting ring, the scraping rod is attached to the inner wall of the elution tank, a cleaning block is arranged on the scraping rod, the cleaning block is in sliding connection with the scraping rod, a rotating ring is arranged below the elution tank, a plurality of groups of annularly distributed bottom plates are arranged below the elution tank, and the plurality of groups of bottom plates are rotationally connected with the elution tank through mounting shafts; the multiple sets of annularly-distributed bottom plates are matched with the scraping rods and the cleaning blocks, so that complete cleaning of kieselguhr on the inner wall of the elution tank is achieved, and the situation that the next use of the eluent recovery device is affected due to the fact that residual kieselguhr cannot be completely cleaned is avoided.

The present invention relates to a hybrid protein comprising a urokinase catalytic domain and a VHH specifically binding to vWF for use in treating a patient having elevated serum lactate dehydrogenase (LDH) levels or at risk thereof following or during a traumatic event. The treatment comprises administering the hybrid protein to reduce the serum LDH levels of the patient to a target range of less than 280 U / L. The present invention also relates to means and methods for the prevention and / or treatment (including, as mentioned herein, reducing and / or limiting the damage caused by) of tissue damage, especially tissue damage associated with the occurrence and / or formation of thrombi in circulation.

The invention relates to the technical field of urokinase, and discloses a urokinase purification method and a chromatographic column type adsorption device for extraction, the purification method comprises the following steps: S1, taking a urokinase raw material, carrying out microfiltration, adding a chitosan quaternary ammonium salt solution, carrying out directional flocculation, centrifuging, and collecting a supernatant; s2, adding the supernate in the step S1 into a weak anion exchange chromatographic column, eluting, collecting eluent with active components, and desalting the eluent; s3, adding the eluent desalted in the step S2 into a gel filtration chromatography column, eluting, and collecting the eluent; s4, the eluent in the step S3 is added to a Ni-IMAC metal chelating chromatographic column, and purified urokinase is obtained after elution. According to the method, multiple chromatographic columns are used for chromatography, and cost and operation complexity are reduced.

Provided herein are methods for safe and effective thrombolysis in therapy for human subjects with symptoms of a potential stroke or acute myocardial infarction (“AMI”) using a sequential administration of a low dose bolus of human tissue plasminogen activator (“tPA”) followed by an infusion of a mutant form of human pro-urokinase (“proUK”).

The invention relates to the technical field of separation and extraction of natural products, in particular to a method for extracting unimodal urokinase from human urine. The method sequentially comprises the following steps: human urine collection, silica gel chromatography, precipitation, aprotinin chromatography, pasteurization for virus inactivation, anisole chromatography, molecular sieve chromatography and freeze-drying. A novel purification process is designed, a specific purification technology is adopted to ensure that a final product shows a single peak in chromatographic analysis, the content of impurity protein in a high-purity product is extremely low, anaphylactic reaction and immunogenicity can be reduced, the medication safety of a patient can be improved, and the method can be directly used for development of a freeze-drying preparation or an injection and simplification of a downstream preparation process. The implementation of the scheme provided by the invention can promote the upgrading of the thrombolytic drug industry chain in China and improve the international competitiveness. The method has the advantages of high finished product purity, few impurities, simplicity and convenience in operation, concise extraction steps, reduced production time, high efficiency, small environmental pollution, low production cost and the like.

This disclosure relates to nanoparticles coated with fusion proteins comprising a domain that binds a cancer marker and a domain comprising a toxic polypeptide. In certain embodiments, the targeted cancer marker is urokinase plasminogen activator receptor (uPAR) insulin-like growth factor 1 receptor (IGF1R), EGFR, HER2, and / or other member of the ErbB family of receptors. In certain embodiments, the molecule that binds a cancer marker is an amino terminal fragment of uPA or variant capable of binding uPAR and / or IGF1 or variant capable of binding IGF1R. In certain embodiments, the toxic polypeptide is a bacterial exotoxin.

Disclosed are novel urokinase plasminogen activator receptor (uPAR) binding molecules and soluble urokinase plasminogen activator receptor (suPAR) binding molecules and methods of their use.

The invention discloses a CHO cell line and a method for producing recombinant human low molecular weight urokinase (rh-LLW-uPA) by using the CHO cell line. The method has high productivity and enables the production of low molecular weight urokinase having improved characteristics in terms of safety, quality and stability.

The application belongs to the technical field of biotechnology, and particularly relates to a method for extracting crude urokinase based on macroporous resin. The method significantly improves the specific activity and shelf life of the crude urokinase through a dynamic pH segmentation protection technology. In the process of lectin affinity chromatography, the high flow rate condition of the first stage pH 8.5-9.5 ensures the efficient and specific binding of urokinase and lectin, and the rapid removal of impure proteins; the dynamic adjustment of the second stage pH from 8.5-9.5 linearly to 7.5-8.5 in combination with low flow rate operation provides a mild microenvironment for urokinase molecules, effectively protecting the integrity of the sugar chain structure of the molecules. The added polyol and disaccharide complex protective agent can stabilize the protein conformation, prevent sugar chain degradation and modification, and reduce the impact of pH mutation on the protein structure. The addition of the complex protective agent further enhances the stability of the sugar chain structure, preventing enzymatic and chemical degradation during the chromatography process.

The present invention relates to the a positron-emitting imaging agent for use in the prognosis of a head and neck cancer (HNSCC) in a subject by PET imaging of the cancer, wherein said imaging agent comprises a uPAR binding peptide coupled via the chelating agent NOTA or DOTA to the radionuclide 68Ga or 64Cu.

The application provides a composite microsphere, a preparation method and application thereof, and a method for purifying urokinase, and belongs to the fields of bioengineering and chemical engineering.The raw material of the composite microsphere comprises modified nanometer ferroferric oxide, polylactic acid, polylysine and arginine.The composite microsphere comprising the modified nanometer ferroferric oxide, the polylactic acid, the polylysine and the arginine is used as a chromatographic column filler, and a chromatographic column is carried out on a urokinase raw material solution, so that the adsorption capacity for endotoxin can be effectively improved, and the activity of the urokinase is improved.

The present invention provides Therapeutic application of 64Cu labelled uPAR binding conjugates for site-specific targeting of the Urokinase Plasminogen Activator Receptor (uPAR) in treatment of cancer diseases associated with high uPAR expression.

The application discloses screening of hemostatic active components in Euphorbiaceae plants and application thereof, and based on ultrafiltration mass spectrometry technology and taking urokinase-type plasminogen activator as a key target enzyme, a hemostatic active component, breynol, of cyclopentanone isocoumarins in Euphorbiaceae plants is screened for the first time. The breynol shows strong specific binding to urokinase plasminogen activator, and the activity inhibition (IC 50 ) of the breynol to the urokinase plasminogen activator is 0.187±0.000 mM, which is superior to that of the positive control tranexamic acid (IC 50 =2.425±0.001 mM). The breynol is reported for the first time in the research of hemostatic activity of Euphorbiaceae plants, and can be used for preparation of daily or clinical hemostatic drugs.

The invention provides urokinase-loaded exosome-like nano-vesicles as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. According to the urokinase-loaded exosome-like nano-vesicle disclosed by the invention, a thrombolytic drug and inherent active ingredients of radix puerariae are packed in one vesicle, so that the urokinase-loaded exosome-like nano-vesicle is obtained; in the urokinase-loaded exosome-like nano-vesicle, active ingredients of radix puerariae and urokinase can generate a synergistic effect, so that deep vein thrombosis is effectively dissolved, meanwhile, the bleeding risk is reduced, the half-life period of urokinase is prolonged, damaged vascular endothelium is repaired in time, the recurrence rate of thrombosis is reduced, and the urokinase-loaded exosome-like nano-vesicle can be used for treating deep vein thrombosis. The invention provides a new direction and strategy for the development of safe deep vein thrombosis treatment drugs, and has good practicability.

The invention belongs to the technical field of biology, and particularly relates to a method for extracting an ulinastatin crude product based on macroporous resin. According to the method, the specific activity and shelf life of the ulinastatin crude product are remarkably improved and prolonged through a dynamic pH segmented protection technology. In the agglutinin affinity chromatography process, high-efficiency specific binding of ulinastatin and agglutinin is ensured under the high flow rate condition of pH 8.5-9.5 in the first stage, and impure protein is rapidly removed; in the second stage, dynamic adjustment of linearly reducing the pH value from 8.5-9.5 to 7.5-8.5 is matched with low-flow-speed operation, so that a mild microenvironment is provided for ulinastatin molecules, and the completeness of a sugar chain structure of the ulinastatin molecules is effectively protected. The added polyhydric alcohol and disaccharide composite protective agent can stabilize protein conformation, prevent sugar chain degradation and modification, and reduce the impact of pH mutation on the protein structure. The addition of the composite protective agent further enhances the stability of the carbohydrate chain structure, and prevents enzymolysis and chemical degradation in the chromatography process.

The invention belongs to the technical field of urine collection, and particularly discloses a self-cleaning urine collection urinal and a use method thereof. A self-cleaning urine collecting urinal comprises a urinal body and a bypass pipe, a water inlet of the urinal body is communicated with a water inlet pipe, a liquid outlet of the urinal body is communicated with a liquid discharging pipe, the bypass pipe and the water inlet pipe are provided with electromagnetic valves, the liquid discharging pipe is provided with a separating and discharging mechanism, and the separating and discharging mechanism is connected with the bypass pipe. A liquid outlet of the liquid discharging pipe is connected with a first liquid inlet of the separating and discharging mechanism, a water outlet of the bypass pipe is connected with a second liquid inlet of the separating and discharging mechanism, and a flushing water discharging opening and a urine discharging opening are formed in the bottom of the separating and discharging mechanism. Through the electromagnetic valve, the bypass pipe and the separating and discharging mechanism, urine collection and automatic and independent discharging of flushing water are achieved, subsequent extraction of urokinase from urine is facilitated, and self-cleaning is also facilitated.

The present invention relates to a method of treating a glomerular kidney disease in a human patient in need thereof, comprising administering to the patient an isolated antibody or an antigen binding fragment thereof that binds to soluble urokinase plasminogen activator receptor (suPAR). The present invention also relates to a method of reducing proteinuria associated with a kidney disease in a patient in need thereof, comprising administering to the patient an isolated antibody or an antigen binding fragment thereof that binds to suPAR. The present invention also relates to a method of inhibiting suPAR in a patient in need thereof, having a glomerular kidney disease, with the same isolated antibody or fragment thereof.

This invention discloses a regenerable urokinase silica gel adsorbent, its preparation method, and its applications, belonging to the field of new materials. The regenerable urokinase silica gel adsorbent is prepared through the following steps: Step 1, preparing a composite template emulsion; Step 2, preparing a sol-gel system; Step 3, preparing a silica gel precursor; Step 4, preparing the regenerable urokinase silica gel adsorbent. This invention avoids the use of expensive chemical ligands and precipitation reagents, and the regeneration process only requires inexpensive buffer solutions without the need for high-concentration salts or denaturants. Therefore, the material and process costs of this invention are significantly reduced compared to existing technologies. Simultaneously, the simplified process greatly reduces water and electricity consumption and chemical waste discharge, providing a solid technical foundation for the large-scale, low-cost, and continuous production of urokinase.