61 results about "Urate oxidase" patented technology

The present invention provides a manganese metal-organic framework material with oxidase properties, a preparation method, and an application thereof. The material is prepared by the following method: using 1,3,5-trimethic acid as an organic ligand, manganese acetate tetrahydrate as a manganese source, and a water-ethanol solution as a solvent, to obtain a manganese metal organic framework (MOF) precursor via a solvothermal synthesis technique; the manganese MOF precursor is post-synthetically modified using an oxidant, wherein the oxidant is a mixed solution of an aqueous alkali metal hydroxide solution and H2O2, and the ratio of the manganese MOF precursor to the alkali metal hydroxide is (400-600) mg:(0.5-2) mmol. By using specific types and amounts of manganese MOF precursors and oxidants, the resulting metal-organic framework nanomaterial has specific enzyme-like properties, can exert the function of uricase, and has optimal and high uricase activity under neutral pH conditions.

The invention provides a manganese metal organic framework material with oxidase characteristics as well as a preparation method and application thereof, and the material is prepared by the following method: taking 1, 3, 5-trimesic acid as an organic ligand, manganese acetate tetrahydrate as a manganese source and a water-ethanol solution as a solvent, and obtaining a manganese MOFs precursor through a solvothermal method synthesis technology; the manganese MOFs precursor is subjected to post-synthesis modification with an oxidizing agent, the oxidizing agent is a mixed solution of an alkali metal hydroxide aqueous solution and H2O2, and the dosage ratio of the manganese MOFs precursor to the alkali metal hydroxide is (400-600) mg: (0.5-2) mmol. The manganese MOFs precursor and the oxidizing agent with specific types and using amounts are adopted, so that the obtained metal organic framework nano material has specific enzyme-like characteristics, can play a role of urate oxidase, has optimal urate oxidase activity under a neutral pH condition, and is relatively high in activity.

The invention provides a human urate oxidase mutant as well as a preparation method and application thereof. The amino acid sequence of the mutant is as shown in SEQ ID NO.1 in a sequence table. The preparation method comprises the following steps: (a) denaturation pretreatment of an inclusion body; (b) denaturation of the inclusion body; and (c) a protein renaturation method is adopted, the preparation process does not need a column chromatography step, the production cost is saved, and the obtained mutant protein is relatively high in purity, has urate oxidase activity, can be used for oxydative degradation of uric acid, and has important clinical application value.

The invention discloses a urate oxidase mutant and application thereof in improving the thermal stability of urate oxidase, and belongs to the technical field of biology. The urate oxidase mutant disclosed by the invention is T68L / T75S / E222D / K299E, T68L / T75S / K299E or T75S / K299E, and the urate oxidase mutant disclosed by the invention is T68L / T75S / K299E. According to the method, the key sites influencing the thermal stability are mined in the modes of directed evolution, high-throughput screening, multiple virtual screening and the like, and the stability is improved through mutation. And finally, a plurality of key sites and mutants influencing the stability are excavated, and the thermal stability is improved.

The invention discloses a urate oxidase mutant and application thereof in improving the heat stability of urate oxidase and the trypsin resistance, and belongs to the technical field of biology. The urate oxidase mutant disclosed by the invention is Y47F / T68L / T75W / D111G / K204F / F259I / K299E, T68L / T75W / K204F / K299E or T68L / T75W / K299E. The invention further discloses a preparation method of the urate oxidase mutant. According to the method, the key sites influencing the thermal stability are mined in the modes of directed evolution, high-throughput screening, multiple virtual screening and the like, and the stability is improved through mutation; the key sites influencing the enzymolysis resistance are excavated through molecular dynamics simulation and solvent accessible surface area calculation, and the enzymolysis resistance effect is improved through mutation. Finally, a plurality of key sites and mutants influencing the stability and the enzymolysis resistance are excavated, and the thermal stability and the enzymolysis resistance are improved.

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.

Provided is a method for improving the stability of urate oxidase. The inventors have discovered that combining an active ingredient urate oxidase or chemically modified urate oxidase with a stabilizer xanthine through a non-covalent bond can significantly improve the in vivo and in vitro stability of urate oxidase and chemically modified urate oxidase in the form of a tetramer, thereby effectively improving the stability of urate oxidase.

The invention provides a dihydrofuro-indole compound with a structure as shown in a formula I, or a derivative thereof, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a solvate thereof, or a prodrug molecule thereof, or a deuteride thereof, or a tritide thereof, and application of the dihydrofuro-indole compound or the derivative thereof, or the stereoisomer thereof, or the pharmaceutically acceptable salt thereof, or the solvate thereof, or the prodrug molecule thereof, or the deuteride thereof, or the tritide thereof. The dihydrofuro-indole compound or the derivative thereof provided by the invention is a compound with a novel structure, and the compound has relatively good inhibitory activity on urate oxidase and urate transporter protein, realizes XOR / URAT1 dual inhibition, has a good uric acid reducing effect, is good in safety, has a very good pharmacokinetic property, and can be used for preparing a medicine for treating uric acid. The compound is high in druggability, can be used for preparing uric acid reducing medicines, and is used for preventing and / or treating gout or hyperuricemia. # imgabs0 #

The invention discloses a uricase mutant. The uricase sequence from aspergillus flavus and the uricase sequence from candida are taken as templates, multiple rounds of random mutation are carried out, sequencing verifies that the key amino acid site of the catalytic center is mutated, and the catalytic activity is enhanced. In-vivo and in-vitro uric acid reducing activities of the two uricase mutants are further verified, and the two uricase mutants can significantly reduce the content of uric acid and are expected to become new uricase drugs for treating hyperuricemia.

The present invention provides novel uricase conjugates comprising a first domain and a second domain. The first domain comprises a uricase polypeptide and the second domain is an unstructured random coil polypeptide domain comprising at least about 100 amino acids. The invention also provides methods of using the uricase conjugates of the invention, e.g., for the treatment of tumor lysis syndrome and / or gout.

The invention belongs to the technical field of uric acid detection, and relates to a preparation method and application of a uric acid and pH dual-electrode detection sensor. The preparation method comprises the following steps: mixing a soluble manganese salt with a complexing agent, adding a soluble hexacyanoferrate solution, aging and standing at room temperature, centrifuging, drying, and pyrolyzing at high temperature to obtain the Mn2O3 / Fe2O3 material. Coating on the surface of a first working electrode, and then continuously dropwise adding a mixed solution containing a cross-linking agent and urate oxidase; and uniformly mixing a chloroauric acid solution with a hydrochloric acid solution and an aniline solution with a hydrochloric acid solution, carrying out electro-deposition on a second working electrode, and drying at low temperature to obtain the sensor. The sensor designed based on the double working electrodes shows excellent stability, reproducibility and anti-interference performance in sweat uric acid monitoring. The sensor can realize rapid identification and dynamic response of uric acid in a real sweat sample, and is expected to be used for integration of wearable equipment to realize continuous gout early warning.

A strain 44XB capable of producing urate oxidase and a use thereof, relating to the technical field of microorganisms and medicines. A strain 44XB capable of producing urate oxidase is preserved in the Korean Collection for Type Cultures (KCTC), with the preservation number: KCTC 43631, the preservation date: 26 September 2023, the preservation address: 181 lpsin-gil. Jeongeup-si, Jeonbuk 56212, South Korea, and the classification name: Ornithinibacillus sp. A use of the strain 44XB in preparation of drugs for treating hyperuricemia and gout. The strain 44XB can produce highly active urate oxidase, and can be used for microecological therapy of hyperuricemia to reduce serum uric acid in hyperuricemia, and thus has the potential to be used in microecological treatment of hyperuricemia and gout.

The application provides a uric acid oxidase secretion signal peptide and application thereof, and relates to the technical field of genetic engineering. In view of the problem that uric acid oxidases reported in the current literature are difficult to secrete in Pichia pastoris, the endogenous signal peptide of Pichia pastoris is screened, a signal peptide with strong secretion capacity is obtained, and the signal peptide is optimized in a targeted manner, a signal peptide mutant with strong secretion capacity is obtained, uric acid oxidase can be better secreted to the extracellular, and then the extracellular enzyme activity of uric acid oxidase is improved.

The application provides an electrochemical uric acid test paper and a uric acid detection method. The electrochemical uric acid test paper has simple structure, low cost, easy manufacturing process, rapid reaction, sensitive detection, wide storage temperature, wide use temperature, wide effective period, accurate result, simple operation, etc. The uric acid detection method distinguishes the test current of ascorbic acid by different time and different excitation signals, subtracts the current signal of ascorbic acid from the current signal of uric acid and ascorbic acid, eliminates the interference of ascorbic acid, is simple to operate and low in cost, does not need to add other material components, does not need the participation of uric acid oxidase and ascorbic acid oxidase, directly distinguishes the current signal by different voltages, is short in test time, reduces the complex data correction process, maintains good repeatability of data, is simple in logic operation, basically has no influence on the repeatability of test signals, and can effectively remove the interference of ascorbic acid.

The application belongs to the technical field of biology and relates to a human uric acid oxidase mutant with weakened immunogenicity and high activity. The human uric acid oxidase mutant with weakened immunogenicity and high activity is obtained by mutating an inactive human uric acid oxidase with an amino acid sequence of SEQ ID NO. 1, and has amino acid substitutions at positions 83, 85, 119, 121, 151, 222, 232, 233, 240 and 252. The human uric acid oxidase mutant has high homology with a theoretical amino acid sequence of human uric acid oxidase, has high activity while reducing immunogenicity, is suitable for long-term application in clinical treatment of hyperuricemia and gout, and can be used for preparing a drug for treating hyperuricemia and gout.

The invention discloses a urate oxidase gene derived from aspergillus cristatus and application thereof, and the urate oxidase gene is characterized in that a natural urate oxidase gene coding region sequence is obtained from the aspergillus cristatus, and the gene is optimized according to the codon preference of escherichia coli to obtain an optimized sequence. Connecting the optimized gene to a carrier through NdeI and BamHI restriction enzyme cutting sites, and constructing a wild type uox recombinant plasmid; the method comprises the following steps: replacing a sequence of uox of aspergillus cristatus for coding 243-site tyrosine (Y) with a sequence for coding lysine (K) through site-specific mutagenesis, constructing mutant Y243K recombinant plasmids, respectively transforming the recombinant plasmids into competent cells, culturing, and adding IPTG to induce target protein expression. And collecting the induced thalli, carrying out ultrasonication and centrifugation to collect the supernatant so as to obtain a crude enzyme, and purifying the crude enzyme through a nickel affinity column so as to obtain the purified UOX protein. Uric acid is used as a substrate to measure the enzyme activity of the purified UOX, and the result shows that the specific enzyme activity of the mutant Y243K is obviously higher than that of a wild type, and the activity is improved by 2.75 times, so that the mutant Y243K has higher application value.

The invention relates to the technical field of protein drugs, in particular to a urate oxidase variant and a glycosylation modification form thereof. An engineered N-glycosylation site is introduced into urate oxidase, and a pair of engineered disulfide bonds is additionally introduced into a tetramer interface, so that the obtained glycosylation modified urate oxidase variant can improve folding stability, in-vivo performance and immunological properties on the basis of maintaining enzymatic activity.

A uric acid-degrading Bacillus, a biological agent and its preparation method. The uric acid-degrading Bacillus is Bacillus sp. XRK-1, with the preservation number: CCTCC M 20232477, the preservation date: December 06, 2023, and the preservation unit: China Center for Type Culture Collection. The present invention also discloses a microbial agent produced by fermenting the Bacillus sp. XRK-1 strain and its preparation method. The uric acid-degrading Bacillus sp. XRK-1, isolated from sunflower discs, has the ability to produce urate oxidase to degrade urea, with the urate oxidase activity as high as 0.103 U / ml, and can efficiently degrade uric acid in vitro. Bacillus sp. XRK-1 grows rapidly, has strong stress resistance and high biological safety; the microbial agent prepared by its fermentation, as well as the preparation method of the microbial agent, can be used for reducing uric acid and has broad application prospects.

The invention discloses a nucleic acid molecule for coding urate oxidase, a related product and an application of the nucleic acid molecule. The invention discloses a nucleic acid molecule for coding urate oxidase. The nucleic acid molecule comprises a nucleotide sequence as shown in SEQ ID NO. 2. According to the invention, a microbe-derived urate oxidase gene sequence for escherichia coli expression is obtained for the first time, and the gene sequence is easy to clone and express and good in repeatability. The invention further discloses a recombinant vector and a bioengineering bacterium for urate oxidase expression, and the obtained urate oxidase protein is high in activity and simple to prepare.

Provided is a method for preparing a polyethylene glycol-modified urate oxidase, at least 11 of the following amino acid sites of the polyethylene glycol-modified urate oxidase have a PEG modification: T1, K3, K4, K30, K35, K76, K79, K97, K112, K116, K120, K152, K179, K222, K231, K266, K272, K285, K291, and K293, and the preparation method includes: performing a coupling reaction between urate oxidase and polyethylene glycol, wherein the polyethylene glycol is provided in the form of an acidic solution, and a molar ratio of the urate oxidase to the polyethylene glycol is 1: (56 to 94), to obtain the polyethylene glycol-modified urate oxidase.

This invention discloses a uric acid-lowering drug composition, which comprises one or more of the following: levocarnitine, levocarnitine derivatives, pharmaceutically acceptable salts of levocarnitine or its derivatives, glucocorticoids, colchicine, nonsteroidal anti-inflammatory drugs, IL-1β inhibitors, drugs that inhibit uric acid production, drugs that promote uric acid excretion, and uricase. The composition has the effects of lowering uric acid, protecting liver and kidney function, and inhibiting leukopenia.

The invention belongs to the technical field of biology, and relates to a human urate oxidase mutant with weakened immunogenicity and high activity. The human urate oxidase mutant with weakened immunogenicity and high activity disclosed by the invention is a human urate oxidase mutant obtained by mutation of inactive human urate oxidase with an amino acid sequence of SEQ ID NO.1; the amino acid sequence of the amino acid sequence is shown as SEQ ID NO.1. The amino acid sequence has 83rd, 85th, 119th, 121th, 151st, 222nd, 232, 233rd, 240th and 252nd amino acid substitutions. The human uricase mutant keeps high homology with the theoretical amino acid sequence of human uricase, has high activity while reducing immunogenicity, is suitable for long-term use in clinical treatment of hyperuricemia and gout, and can be used for preparing drugs for treating hyperuricemia and gout.

The application belongs to the technical field of biology and relates to a high-activity and high-humanization rate uric acid oxidase mutant. The high-activity and high-humanization rate uric acid oxidase mutant is a human uric acid oxidase mutant obtained by mutation of an inactivated human uric acid oxidase with an amino acid sequence of SEQ ID NO. 1, which has amino acid substitutions at positions 83, 119, 121, 151, 222, 232, 233, 240 and 252, also has amino acid substitutions at positions 208 and 219, and further has an amino acid substitution at position 112. The high-activity and high-humanization rate uric acid oxidase mutant has high homology with a theoretical amino acid sequence of human uric acid oxidase, has high activity, is more suitable for long-term use in the treatment of hyperuricemia and gout in the clinic, and can be used for preparing a drug for treating hyperuricemia and gout.

The present invention provides a dihydrofuroindole compound having a structure as represented by formula I or a derivative thereof, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a solvate thereof, or a prodrug molecule thereof, or a deuteride thereof, or a tritide thereof, and a use thereof. The dihydrofuroindole compound or the derivative thereof provided by the present invention is a compound having a novel structure. The compound has good inhibitory activity against both urate oxidase and a urate transporter, is an XOR / URAT1 dual inhibitor, has a good urate-lowering effect, good safety, good pharmacokinetic properties, and high druggability, can be used for preparing a urate-lowering medicament, and can be used for preventing and / or treating gout or hyperuricemia.

The invention provides a urate oxidase secretion signal peptide and application thereof, and relates to the technical field of gene engineering. Aiming at the problem that urate oxidase is difficult to secrete in pichia pastoris reported in current literatures, endogenous signal peptides of pichia pastoris are screened to obtain signal peptides with relatively strong secretion ability, and the signal peptides are specifically optimized to obtain a signal peptide mutant with relatively strong secretion ability; the urate oxidase can be better secreted out of cells, so that the extracellular enzyme activity of the urate oxidase is improved.

The invention belongs to the technical field of biology, and relates to a high-activity human urate oxidase mutant with immunogenicity weakening. The human urate oxidase mutant with high activity and immunogenicity weakening, disclosed by the invention, is a human urate oxidase mutant obtained by mutation of inactive human urate oxidase with an amino acid sequence of SEQ ID NO.1; the amino acid sequence of the human urate oxidase mutant is SEQ ID NO.2, and the human urate oxidase mutant has 83rd, 85th, 119th, 121st, 151st, 208th, 219th, 222nd, 232, 233rd, 240th and 252nd amino acid substitutions. The human uricase mutant keeps high homology with the theoretical amino acid sequence of human uricase, has high activity while reducing immunogenicity, is suitable for long-term use in clinical treatment of hyperuricemia and gout, and can be used for preparing drugs for treating hyperuricemia and gout.

The invention discloses a high-sensitivity electrochemical luminescence sensor for detecting uric acid as well as a preparation method and a detection method thereof, and belongs to the technical field of detection. A zirconium-based metal organic framework (Zr-TPE MOF) and urate oxidase (UOx) are sequentially dispensed on the surface of a working electrode to serve as modification materials, the Zr-TPE MOF realizes luminous intensity multiplication in an aggregation state through an intramolecular motion limitation (RIM) mechanism, and UOx can react with a to-be-detected sample uric acid in a buffer solution to generate H2O2; the latter can co-react to promote redox electron transfer on the surface of the competitive electrode so as to influence luminescence. The luminescent material based on the novel tetraphenyl ethylene (TPE) derivative has the advantages of high sensitivity, low background signal, strong specificity, simplicity and convenience in operation and the like, and has obvious advantages compared with a previously developed sensor for detecting uric acid. According to the invention, quantitative detection of uric acid can be realized, and an innovative solution is provided for clinical high-sensitivity uric acid detection and early warning of metabolic syndrome.

The present invention relates to the field of sensor analysis and detection technology, and in particular to a self-powered sensor and its preparation method and application. A dual-MOF-derived iron-doped carbon nanotube catalyst is prepared by a solvothermal method and high-temperature calcination. The catalyst is used for the cathode of a zinc-air battery to show excellent power density, energy density, and durability, with performance superior to that of a commercial Pt / C catalyst. Urate oxidase UAO is loaded on the cathode catalyst to form a cathode sensing interface. UAO consumes oxygen to catalyze the oxidation of uric acid, competing with the oxygen reduction process of the zinc-air battery cathode, resulting in a decrease in the output signal. A zinc-air battery self-powered electrochemical sensor based on an oxygen competition model is constructed for uric acid detection. The present invention broadens the application of zinc-air batteries in self-powered sensors, and the constructed sensor has a wide linear range, high sensitivity, and low detection limit for uric acid detection.

Provided is an mRNA-liposome complex. The mRNA-liposome complex includes a liposome and a nucleic acid. The nucleic acid includes at least one mRNA encoding urate oxidase or recombinant urate oxidase. The liposome includes at least two lipids, and the at least two lipids include a core lipid. The liposome has a pKa ranging from 6.0 to 6.3. A molar percentage of the core lipid in a total molar amount of the liposome is not less than 15%.

The present application belongs to the field of biotechnology, and relates to a human urate oxidase mutant with high activity and weakened immunogenicity. The human urate oxidase mutant with high activity and weakened immunogenicity is obtained by mutation of an inactive human urate oxidase with an amino acid sequence of SEQ ID NO. 1; the human urate oxidase mutant with an amino acid sequence of SEQ ID NO. 2 has amino acid substitutions at positions 83, 85, 119, 121, 151, 208, 219, 222, 232, 233, 240 and 252. The human urate oxidase mutant of the present application has high homology with a theoretical amino acid sequence of human urate oxidase, has high activity while reducing immunogenicity, is suitable for long-term application in clinical treatment of hyperuricemia and gout, and can be used for preparing a drug for treating hyperuricemia and gout.