31 results about "Chymase" patented technology

The invention relates to a medicine for skin regeneration and repair and a preparation method thereof, and belongs to the technical field of biological medicine, the medicine comprises cyperus esculentus extract peptide, brooklet anemone root tremella extract, rhodomyrtus tomentosa root-suberect spatholobus stem-radix cyathulae composite extract, pomegranate peel polyphenol and the like. The cyperus esculentus extracted peptide is a product which is more than 1kDa and is obtained by performing enzymolysis on cyperus esculentus through cellulase, alpha-chymotrypsin and ficin protease in sequence and then performing purification through a G-50 sephadex column; the tremellodon gelatinosum tremella extract is a product which is less than 3kDa and is obtained by performing common enzymolysis on tremellodon gelatinosum tremella sporocarp through cellulase and bromelain; the rhodomyrtus tomentosa root-suberect spatholobus stem-radix cyathulae composite extract is a product obtained by performing gradient extraction on rhodomyrtus tomentosa root, suberect spatholobus stem and radix cyathulae through an ethanol water solution to obtain an extract, and performing polyamide resin column adsorption and LH-60 sephadex column purification in sequence. A good repairing system is constructed by multiple components, and wound repairing and healing are accelerated.

The invention discloses a urate oxidase mutant (rAfUOX) with trypsin and chymotrypsin tolerance and application of the urate oxidase mutant (rAfUOX). The invention provides a urate oxidase mutant with trypsin and chymotrypsin tolerance by modifying a key amino acid residue in a wild type aspergillus flavus urate oxidase molecule through a protein engineering technology. The activity half-life period of the recombinant aspergillus flavus urate oxidase mutant rAfUOXK49Q after trypsin digestion is increased by 244% compared with the activity half-life period of the wild type rAfUOX, and the activity half-life period of the recombinant aspergillus flavus urate oxidase mutant rAfUOXK49Q after chymotrypsin digestion is increased by 191% compared with the activity half-life period of the wild type rAfUOX. Other enzymatic properties are basically consistent with those of the wild type rAfUOX.

The disclosure relates to pharmaceutical formulations and method of using chymase inhibitors for the prevention and treatment of mast cell-related disease states in which chymase is involved. More particularly the present disclosure is in the field of treating or preventing or reducing the symptoms of mast cell-related diseases, including atopic dermatitis (AD), chronic urticaria (CU), Eosinophilic Gastrointestinal Disorders (EGIDs), and prurigo nodularis (PN), using a chymase inhibitor, including a chymase inhibitor that prevents cleavage of stem cell factor (SCF) and its interaction with receptor tyrosine kinase (c-kit), thereby reducing mast cell activation and proliferation.

The application discloses a kind of low-toxicity broad-spectrum antibacterial peptide with WW as symmetry center, the antibacterial peptide is that WK is repeated twice and symmetrically distributed in two sides, and WW is used as symmetry center, obtain antibacterial peptide WKWKWWKWKW-NH2 (N2W2), then the tryptophan of C terminal and N terminal of N2W2 is replaced by leucine and / or isoleucine;The antibacterial peptide can basically maintain the excellent antibacterial activity of parent peptide, and their hemolytic activity is greatly reduced, and the safety is improved. The antibacterial activity of IL-N2W2 is highest, but the stability needs to be improved, so D-type amino acid replacement is carried out on IL-N2W2 to obtain D-IL-N2W2. D-IL-N2W2 maintains the high antibacterial activity and low toxicity of IL-N2W2, and can still completely inhibit bacterial growth after incubation with high-concentration trypsin or chymotrypsin, has very great potential in relieving antibiotic resistance, and has good application prospect in preparing clinical antibacterial drugs.

The present invention covers the use of chymase inhibitors in general and more in particular substituted bicyclically substituted uracils of general formula (I) as described and defined herein, and 3-methylbenzo-[b]thiophene)-2-sulfonamido derivatives of general formula (II) for manufacturing pharmaceutical compositions for the treatment or prophylaxis of stroke, pulmonary embolism, deep or superficial vein thrombosis, thrombotic microangiopathy, thrombotic microangiopathy in hypercoagulable states after infection, inflammation, transplantation, disseminated intravascular coagulation, vaccine-induced immune thrombotic thrombocytopenia, vascular access site thrombosis or occlusion.

The application provides a polypeptide with angiotensin converting enzyme inhibitory activity and a preparation method and application thereof, and relates to the technical field of active peptide preparation. The abalone is pulverized, a complex enzyme is added for primary enzymolysis, and then a specific protease mixture containing prolyl endopeptidase, chymotrypsin and protease K is added for two-step enzymolysis, so that the obtained polypeptide solution has high ACE inhibitory activity. The obtained polypeptide is analyzed and screened to obtain polypeptides with amino acid sequences as shown in SEQ ID NO. 1-5. The polypeptide has high ACE inhibitory activity, and the IC 50 0.05mg / mL-0.40mg / mL, and therefore can play a blood pressure lowering effect as an angiotensin converting enzyme inhibitor.

The invention belongs to the technical field of comprehensive utilization of oyster mushrooms, and provides an oyster mushroom active peptide for inhibiting foaming of macrophages and a preparation method of the oyster mushroom active peptide. The method comprises the following steps: taking oyster mushroom as a raw material, preparing oyster mushroom protein through alkali extraction and acid precipitation, preparing a crude product of the oyster mushroom active peptide for inhibiting foaming of macrophages through compound enzymatic hydrolysis of the oyster mushroom protein by pepsin, trypsin and chymotrypsin, and separating and purifying the crude product by adopting ultrafiltration membrane ultrafiltration and Sephadex G-15 gel column chromatography to obtain the peptide with good activity and high purity. The bioactive peptide can reduce lipid accumulation, ROS level, TNF-alpha content and IL-6 content in RAW264.7 foamed macrophages, and effectively improve lipid accumulation, oxidative stress and inflammatory state of the foamed macrophages, thereby inhibiting foaming of the macrophages.

The invention discloses a micro-ecological protective oral cleaning composition for whole-course control of dental calculus and application of the micro-ecological protective oral cleaning composition, and belongs to the technical field of oral care products. The composition contains citrate, a multienzyme mixture (at least two of lysozyme, xylanase and chymotrypsin) and a matrix additive. Citrate is chelated with calcium ions to soften and dissolve calcified dental calculus; enzymolysis of a multi-enzyme mixture inhibits early bacterial attachment and sediment accumulation and cooperates with citrate to decompose calcified dental calculus; the matrix additive ensures stable release of active ingredients. The components are synergistic, the whole process from the initial stage of dental calculus formation to calcification maturation intervenes, inhibits deposition and dissolves a calcification structure, and meanwhile, the composition can protect the micro-ecology of the oral cavity and has no obvious destructive effect on oral flora. The composition is applied to preparation of oral care products, can solve the problems of one-sided cleaning effect, poor hidden area effect and dysbacteriosis caused by sterilization of existing products, achieves the dual purposes of whole-course dental calculus control and micro-ecological maintenance, and is wide in application prospect.

The invention relates to the technical field of antibacterial peptide application, in particular to an antibacterial peptide CAMP502NC3 with high stability and application of the antibacterial peptide CAMP502NC3. The compound not only shows good antibacterial activity on staphylococcus aureus, but also can be applied to wide pH environments with pH of 2-8 and various protease environments, such as pepsin, trypsin and chymotrypsin; the compound can maintain a certain bactericidal ability under the conditions of high temperature and high salt concentration as high as 250 mM, shows high stability, and has the potential of preparing environment-stable antibacterial drugs.

Provided herein are compositions, methods, and kits related to chymotrypsin production for characterizing proteins, assessing, and monitoring disease stages and phases, predicting the likelihood of disease progression, predicting and monitoring responses to disease therapies, and treating disease conditions.

The present invention covers the use of chymase inhibitors in general and more in particular substituted bicyclically substituted uracils of general formula (I) as described and defined herein, and 3-methylbenzo-[b]thiophene)-2-sulfonamido derivatives of general formula (II) for manufacturing pharmaceutical compositions for the treatment or prophylaxis of stroke, pulmonary embolism, deep or superficial vein thrombosis, thrombotic microangiopathy, thrombotic microangiopathy in hypercoagulable states after infection, inflammation, transplantation, disseminated intravascular coagulation, vaccine-induced immune thrombotic thrombocytopenia, vascular access site thrombosis or occlusion.

The invention belongs to the technical field of biology, and particularly relates to separation and application of proteasome-rich human placenta-derived vascular stem cell outer vesicles. The extracellular vesicles provided by the invention are derived from human placenta source blood vessel promoting stem cells, and the human placenta source blood vessel promoting stem cells show that markers PAI-1 are positive, MECOM is positive, CD201 is positive, CD44 is positive, CD73 is positive, HLA-ABC is positive, CD34 is negative, CD31 is negative and CD326 is negative. Compared with human umbilical cord mesenchymal stem cell outer vesicles, the human placenta-derived vascular stem cell outer vesicles provided by the invention are rich in proteasomes and angiogenic protein factors, wherein the proteasomes have high chymotrypsin-like activity. Moreover, in-vivo and in-vitro experimental data show that the human placenta-derived vascular stem cell outer vesicles provided by the invention are rich in proteasomes, and can degrade accumulation of HIF and inhibit cell apoptosis; the medicine has a treatment effect on myocardial infarction.

The present application relates to the technical field of antibacterial peptide application, and particularly relates to an antibacterial peptide CAMP502NC3 with high stability and application thereof.The antibacterial peptide CAMP502NC3 not only exhibits good antibacterial activity on staphylococcus aureus, but also can maintain certain bactericidal capacity in a wide pH environment of pH 2-8, a variety of protease environments, for example, pepsin, trypsin, chymotrypsin, and a salt concentration condition of up to 250 mM, and shows high stability and has the potential to prepare environment-stable antibacterial drugs.

The present application belongs to the field of bioactive peptides. The present application provides an angiotensin converting enzyme inhibitory peptide and a screening method and application thereof, and the amino acid sequence of the inhibitory peptide is GKGLW and / or GDGLKW. The screening method of the inhibitory peptide of the present application uses soybean protein as raw material, adopts targeted alkaline protease and chymotrypsin double enzymolysis technology for preparation and screening, realizes rapid screening of ACE inhibitory peptides, and provides experimental basis for structure-activity relationship. The inhibitory peptide has strong blood pressure lowering activity, high safety and no toxic side effects. The determination of the stability of the digestive tract shows that the inhibitory peptide has good gastrointestinal stability in the gastrointestinal digestion, and can still maintain good activity after entering the blood.

The application discloses a kind of central symmetry β-hairpin antibacterial peptide with KW repeat sequence with R n As corner, and application thereof, the antibacterial peptide is with R, RR or RRR as corner, WK repeat unit is alternately arranged in two sides, its structure general formula is (WK) x R n (KW) x -NH2, marked as R n W x ;Wherein, n=1,2,3;X=2,3.In vitro antibacterial experiment and hemolytic activity experiment show that this kind of antibacterial peptide has strong antibacterial activity and almost negligible hemolytic toxicity.After D-type amino acid full replacement of R2W3 sequence, the antibacterial activity of D-R2W3 is further enhanced, the hemolytic toxicity is low, and it can resist the hydrolysis of trypsin and chymotrypsin, has very great potential in relieving antibiotic resistance, and has good application prospect in preparing clinical antibacterial drugs.

The invention discloses a fluorescence sensing kit for detecting chymotrypsin activity and application thereof, and belongs to the technical field of biology. On the basis of copper ions and hydrogen peroxide, a non-fluorescent substance o-phenylenediamine can be slowly oxidized into a fluorescent product 2, 3-diaminophenazine, after the copper ion-casein compound is introduced, the compound can form mimetic peroxidase, the mimetic peroxidase and the hydrogen peroxide are combined to accelerate the reaction rate, and the fluorescence intensity is remarkably enhanced. The chymotrypsin can shear a substrate casein, so that a copper ion-casein compound is fragmented, the combination effect of the copper ion-casein compound is changed, the catalytic activity of the compound is influenced, generated fluorescent products are reduced, and the fluorescence intensity is reduced, a chymotrypsin activity detection method is established, and the activity of the chymotrypsin is determined by detecting the fluorescence intensity. According to the method, chymotrypsin activity detection and inhibitor screening can be efficiently and accurately achieved, and a new method is provided for rapid detection of chymotrypsin activity.

The invention belongs to the technical field of experimental model preparation, and provides a construction method of an in-vitro articular cartilage superficial layer injury model aiming at proteoglycan and collagenous fibers in a superficial cartilage layer (SFZ) in order to simulate cartilage surface component loss and structural damage in the early stage of OA morbidity, and the in-vitro articular cartilage superficial layer injury model is constructed by taking a pig knee joint cartilage as a material. A mixed solution of 0.01 mg / ml chymotrypsin and 0.5 mg / ml collagenase D is used as digestive enzyme, the surface of an in-vitro articular cartilage column is treated for 20 min, proteoglycan and collagenous fibers in SFZ are accurately destroyed, and an in-vitro articular cartilage superficial layer damage model is constructed; histological staining, microscopic observation and mechanical detection results all show that the method can cause SFZ structure damage and mechanical property reduction on the premise of not influencing deep tissues. The model has the advantages of simplicity and convenience in operation, low cost and controllable injury range, and provides a reliable experimental tool for research on early pathological mechanisms of osteoarthritis and exploration of related diagnosis and treatment means.

The application discloses a preparation method of balsam pear seed peptides by enzymolysis, and belongs to the technical field of peptide preparation. The preparation method comprises the following steps: after balsam pear seeds are crushed, water is added for soaking treatment to obtain material A; cellulase and pectinase are added to the material A for pretreatment, enzyme inactivation is performed, and a primary enzymolysis liquid is obtained; compound protease is added to the primary enzymolysis liquid for step-by-step enzymolysis, enzyme inactivation is performed, solid-liquid separation is performed, and an enzymolysis liquid is obtained; the enzymolysis liquid is dried to obtain polypeptide powder; the polypeptide powder is extracted by ethanol, filtered, and dried to obtain the balsam pear seed peptides; and the compound protease in step (3) is bromelain, pepsin and carboxypeptidase B. Compared with the prior art, the balsam pear seed peptides prepared by the application have better alpha-glucosidase inhibiting capacity, are close to chemical hypoglycemic drugs, can be used as natural alpha-glucosidase inhibitor raw materials, and have lower side effects.

The invention provides chymotrypsinogen and application thereof, and particularly relates to a protein tag for improving the activation efficiency of recombinant chymotrypsinogen. Compared with naturally extracted human chymotrypsinogen, the recombinant chymotrypsinogen fused with the protein tag has significantly improved activation efficiency. The invention also relates to a recombinant chymotrypsinogen containing the protein tag, a nucleic acid molecule for coding the recombinant chymotrypsinogen, a vector containing a nucleotide sequence of the nucleic acid molecule, a nucleic acid construct and a host cell. In addition, the invention also relates to a method for expressing the recombinant chymotrypsinogen.

The invention relates to a fluorescent probe for protease activity detection, and particularly discloses a fluorescent probe DCD-7F for detecting chymotrypsin activity as well as a preparation method and application of the fluorescent probe DCD-7F. The invention relates to a fluorescent probe for detecting chymotrypsin activity, which uses a DCDNH2 luminophore as a parent nucleus structure of ratio fluorescence signal change and heptaflutolanamide in DCD-7F as a recognition group to replace a traditional ester group low-specificity probe hydrolysis site. Spectrum test screening finds that the DCD-7F probe has high specificity and selectivity when being used for chymotrypsin activity detection. Based on excellent ratio spectral characteristics of DCD-7F in detection of chymotrypsin activity, the DCD-7F is applied to real-time detection of chymotrypsin activity in a pancreatitis animal model, and early diagnosis, prevention and treatment of pancreatitis can be effectively assisted by reasonably applying a DCD-7F fluorescence imaging technology.

The present invention provides a Golgi-targeted near-infrared fluorescent probe for detecting chymotrypsin, as well as its preparation method and application. The structural formula is as follows: #imgabs0#, wherein R is selected from #imgabs1#. The probe of the present invention uses a hemicyanine derivative as a parent nucleus, introduces p-aminobenzenesulfonamide as a Golgi-targeting group, and uses 4-bromobutyrate or 3-phenylpropionate as a recognition group for detecting chymotrypsin. Chymotrypsin-induced ester group cleavage is accompanied by a significant enhancement of the fluorescent signal. The probe can effectively detect chymotrypsin from different proteases, amino acids, and yin and yang examples, with good selectivity and high sensitivity. The probe is accompanied by a distinct color change visible to the naked eye when acting on chymotrypsin, and exhibits good Golgi targeting capability, resulting in successful applications for chymotrypsin detection and imaging in biological systems.

The present application relates to a kind of skin regeneration repair medicine and preparation method thereof, belong to biological medicine technical field, and medicine includes oil palm extraction peptide, tiger palm silver ear extract, Gangdeng root-milletia nitida-Siberian milkvetch composite extract and pomegranate peel polyphenol etc.Oil palm extraction peptide is oil palm sequentially by cellulase, alpha-chymotrypsin and ficin enzymolysis, then by G-50 dextran gel column purification and obtain the product above 1kDa;Tiger palm silver ear extract is tiger palm silver ear fruit body by cellulase and bromelain common enzymolysis, and obtain the product below 3kDa;Gangdeng root-milletia nitida-Siberian milkvetch composite extract is Gangdeng root, milletia nitida and siberian milkvetch by gradient extraction of ethanol aqueous solution and obtain extract, sequentially by polyamide resin column adsorption and LH-60 dextran gel column purification and obtain the product.Multiple components construct good repair system, accelerate wound repair healing.

A method for screening and preparing antioxidant peptides from mare's milk whey protein and uses thereof. Potential antioxidant polypeptides are screened based on in silico enzymolysis and computer-based prediction, followed by molecular docking with the keap1 protein. The feasibility of obtaining antioxidant peptides is then validated by simulating gastrointestinal digestion using pepsin, trypsin, and chymotrypsin. Subsequently, high-purity antioxidant peptides are chemically synthesized and subjected to in vitro chemical antioxidant activity assays.

The present invention provides a class of HSA fluorescent probes and their synthesis and applications. The fluorescent probe is an enzymatic HSA fluorescent probe having a structure represented by formula (I). The fluorescent probe of the present invention has high selectivity and high sensitivity, capable of distinguishing HSA from other enzymes (such as chymotrypsin, lipase, protease, β-galactosidase, pepsin, acetylcholinesterase, butyrylcholinesterase, carboxylesterase, trypsin, DNA enzyme, alkaline phosphatase, lysozyme, etc.), with a detection limit as low as 0.27 μg / mL. Therefore, the fluorescent probe of the present invention can achieve qualitative and quantitative detection of HSA in serum, urine, or cells.

Provided herein are methods of making a peptide aptamer library of partially random peptide aptamers. The peptide aptamer library can be designed with tryptic and chymotryptic sites spaced to create domains. In each domain, a defined subset of amino acids are randomly ordered alongside invariant amino acids. Also provided herein are methods to identify peptide aptamers that bind a target using the peptide aptamer library disclosed herein.

The invention provides a marine bioactive polypeptide as well as preparation and application thereof, and relates to the technical field of bioactive peptide preparation. According to the method, the abalone is crushed, the compound enzyme is added for primary enzymolysis, then the specific protease mixture containing prolyl endopeptidase, chymotrypsin and protease K is added for secondary enzymolysis, and the obtained polypeptide liquid has high ACE (angiotensin converting enzyme) inhibition. The obtained polypeptide is analyzed and screened to obtain the polypeptide with the amino acid sequence as shown in SEQ ID NO.1-5. The polypeptide disclosed by the invention has high ACE (angiotensin converting enzyme) inhibition, and IC50 (50% of IC50) of the polypeptide to ACE is 0.05 mg / mL-0. 40mg / mL, so that the polypeptide can be used as an angiotensin converting enzyme inhibitor to play a role in reducing blood pressure.

Provided are a chymotrypsinogen and the use thereof. The present invention specifically relates to a protein tag capable of improving the activation efficiency of a recombinant chymotrypsinogen. Compared with naturally extracted human chymotrypsinogen, the recombinant chymotrypsinogen in which the protein tag is fused has a significantly improved activation efficiency. The present invention also relates to a recombinant chymotrypsinogen comprising the protein tag, a nucleic acid molecule encoding the recombinant chymotrypsinogen, a vector comprising the nucleotide sequence of the nucleic acid molecule, a nucleic acid construct and a host cell. In addition, the present invention also relates to a method for expressing the recombinant chymotrypsinogen.

The invention relates to an extraction method of a micromolecular protein compound and application of the micromolecular protein compound in the field of skin care. According to the method, a goat embryo is taken as a raw material, and a protein compound taking Ubiquitin as a main material and EFS, ALB, BIN1, H2AJ, FBLN1 and KPNA3 as auxiliary materials is obtained through tissue crushing, chymotrypsin hydrolysis and hydrophobic chromatography enrichment. The compound has hydrophobic and hydrophilic amphoteric characteristics and is used in skin care products to realize skin barrier penetration and activity exertion.