78 results about "Nucleotidase" patented technology

Compounds that modulate the conversion of AMP to adenosine by 5′-nucleotidase, ecto, and compositions containing the compounds and methods for synthesizing the compounds, are described herein. The use of such compounds and compositions for the treatment and / or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, that are mediated by 5′-nucleotidase, ecto is also provided.

The invention relates to the field of an external diagnostic reagent, and particularly relates to a high-sensitivity kit for detecting 5'-nucleotidase. The kit for detecting 5'-nucleotidase comprises a 5'-nucleotidase R1 reagent, a 5'-nucleotidase R2 reagent and a 5'-nucleotidase calibration product, wherein the 5'-nucleotidase R1 reagent comprises a buffer solution, a stabilizing agent, a preservative agent, an enzyme activator, 4-AAP(4-aminoantipyrine), 18-crown ether-6 and UAO (Uricase), PNP (Purine Nucleoside Phosphorylase), XOD (Xanthine Oxidase) and POD (Peroxidase) reaction enzyme systems; the 5'-nucleotidase R2 reagent comprises a buffer solution, a stabilizing agent, a preservative agent, PO3- and a Trinder developing system; and the 5'-nucleotidase calibration product comprises a buffer solution, a stabilizing agent, a preservative agent and 5'-nucleotidase. The detection kit provided by the invention is enhanced in detection sensitivity by 40% compared with a 'PNP-XOD-POD' enzyme system, and has good application prospects.

The invention relates to treating a disease or condition in which it is desirable to inhibit 5′-methylthioadenosine phosphorylase (MTAP) and / or 5′-methylthioadenosine nucleosidase (MTAN). The invention particularly relates to the co-administration of 5′-methylthioadenosine (MTA), or a prodrug of MTA, with one or more MTAP / MTAN inhibitors. Included among the diseases treatable are prostate cancer and head and neck cancer.

This invention relates to antibodies that bind an epitope present on CD73 expressed at the surface of cells, including tumor cells, and that inhibit the enzymatic (ecto-5′ nucleotidase) activity of the CD73 enzyme. Such agents can be used for the treatment of diseases such as cancers.

This disclosure relates to antibodies that bind an epitope present on CD73 expressed at the surface of cells, including tumor cells, and that inhibit the enzymatic (ecto-5′ nucleotidase) activity of the CD73 enzyme. Such agents can be used for the treatment of diseases such as cancers.

The invention provides a fermentation method for preparing nuclease P1, which using directly penicillium citrinum sporogon as seed, fermenting by means of fed-batch sugar supplement to improve activity of the nuclease P1, the mehod without first and second order seed culture shorts production cycle greatly and raises efficiency. The invetion also uses obtained enzyme for preparing 5'-mononucleotide with 85-90% rate of enzymatichydrolysis and 15 mg / ml of producing rate of nucleotide. The invention also provides a penicillium citrinum 3.2788-Y01 of nuclease P1, which fungus stroing number is CGMCC NO.1943, the stem can raise one times active unit of the nuclease P1.

The present invention provides ectonucleotidase inhibitors represented by the following formula, including ecto-nucleotide triphosphate diphosphohydrolase (NTPDase) inhibitors and ecto-5′-nucleotidase (ecto-5′-NT) inhibitors, namely nucleotide mimetics as selective NTPDase or ecto-5′-NT inhibitors. It also provides methods for preparations of said compounds. Furthermore provided are pharmaceutical and diagnostic compositions comprising said compounds, and the use of said compounds in a medicament for treating diseases associated with ectonucleotidase activity and / or P1 or P2 receptors.

The present invention relates to a microorganism producing inosine, which is one of purine nucleoside, an important material for 5′-inosinic acid synthesis, and method for producing inosine using the same. More particularly, the present invention relates to a recombinant microorganism of Corynebacterium genus producing inosine at high concentration by inactivating the gene encoding nucleoside hydrolase II and by enhancing the expression of the gene encoding 5′-nucleotidase, which still retains the characteristics of Corynebacterium ammoniagenes CJIP2401 (KCCM-10610).

The invention relates to a method for determining activity of 5′-nucleotidase, in which a biological sample is incubated in a manner and under conditions known per se with a nucleotide pentose monophosphate as substrate, the liberated inorganic phosphate is converted into a colored complex by treating it with ammonium molybdate and a reducing agent, color intensity is measured by a known method, and from the measured value the activity of 5′-nucle-otidase or the amount of inorganic phosphate liberated within unit time, as a figure proportional to the activity of 5′-nucleotidase, is calculated with a known calculation and / or with the aid of a calibration curve. According to the invention 5′-AMP, 5′-CMP, 5′-UMP, 5′-GMP, 5′-IMP and 5′-TMP are used as nucleotide pentose monophosphates, and measurement is performed on all of these six substrates. The invention also relates to a reagent kit for performing the above method.

The invention relates to a method for decolorizing nucleotide enzymatic hydrolysate. The method comprises the following steps of: performing pigment adsorption on the nucleotide enzymatic hydrolysate through hyper-crosslinked resin SX-01; and after adsorption is saturated, and regenerating the hyper-crosslinked resin SX-01 by using a regenerant, wherein the hyper-crosslinked resin SX-01 has the following structure units: the skeleton of the resin is polystyrene, the average grain size is 0.3 to 1 mm, the water content is 32 to 49 percent, the aperture is 1.5 to 4 nm, the most probable aperture is 1.8 nm, the average specific surface area is 950 to 1,800 m<2> / g, and the average pore volume is 0.54 to 0.82 cm<3> / g. By adoption of the method, the decolorization ratio of the enzymatic hydrolysate reaches over 80 percent, the treatment quantity reaches 11 times of bed volume (BV), the loss ratio of four kinds of nucleotides is lower than 5 percent, and the consumption quantity of the regenerant is 5 BV.

The invention relates to a 5'-nucleotidase diagnosis reagent box of Enzymatic Recycling Method, and the invention also relates to method principle for detecting the solution of 5'-nucleotidase, constitute and component of reagent, which belongs to the technical field of medical checking measurements. The reagent box in the invention can be dry powder state, and used after dissolution; it also can be formulated to be liquid agent for direct usage. The component of reagent box mainly contains: buffering liquid, alpha- ketoglutaric acid, reduction type coenzyme, Adenosine Deiminase EC 3.5.4.4, glutamate dehydrogenase EC 1.4.1.2, EC 1.4.1.3, EC 1.4.1.4, Glutamate oxidase EC 1.4.3.7, EC 1.4.3.11, peroxidase EC 1.11.1.7, reduction type chromogen assembly, stabilizer and anti-interference agent; and the component can be mixed to form single-reagent reagent box, two-reagent reagent box, and three-reagent reagent box; by series of enzymatic reaction, the colorless reduction type chromogen assembly is oxygenated to form color dye, and the content of dye can be measured by visible light analyzer at wavelength of 400-700nm to reflect the concentration of 5'-nucleotidase directly. Comparing with present technique, the invention can be spread easily, and because the produced dye has higher molar extinction coefficient, the sensitivity is high, and the precision is well.

The invention discloses a xanthine dehydrogenase intercepting body and an application thereof. The protein comprises xanthine dehydrogenase intercepting body small subunit, xanthine dehydrogenase intercepting body intermediate subunit, and xanthine dehydrogenase intercepting body large subunit. The affinity of xanthine dehydrogenase intercepting body is increased by 19% by comparing a wild substrate (xanthine), the conversion number is increased by 115%, the catalysis efficiency is increased by 166%, and the temperature toleration is increased by 11 DEG C. The xanthine dehydrogenase intercepting body is in favor of developing the novel application field by combining with other enzymes, and is suitable for sample detection for detecting xanthine and hypoxanthine, the xanthine dehydrogenase intercepting body can be used for detecting inorganic phosphoric acid by combining with PNP enzyme, can be used for detecting adenosine deaminase by combining with PNP, XOD and POD enzymes and detecting 5'-nucleotidase, and is suitable for industrial application. The xanthine dehydrogenase intercepting body is in favor of increasing enzyme catalysis efficiency and reducing cost, and is in favor of industrial application.

Aortic valve stenosis (AS) is a chronic process related to a progressive mineralization of the aortic root and valve cusps. We found in human AS valves a high level of expression and enzymatic activity of ectonucleotide pyrophosphatase / phosphodiesterase-1 (ENPP-1), which correlated to the degree of mineralization. In vitro, inhibition of ENPP activity with ARL 67156 significantly reduced calcification of isolated valve interstitial cells. In a rat model of cardiovascular calcification, ARL 67156 significantly reduced calcification of the aortic root and valve cusps. This is the first study to demonstrate that increased expression and activity of ENPP-1 promotes the mineralization process in AS valves. Hence, inhibition of ectonucleotidase may represent a novel target of therapy for this frequent and serious cardiovascular disease.

The invention provides a recombinant microorganism for producing beta-nicotinamide mononucleotide (NMN) and a method for producing NMN by using the recombinant microorganism, the strain of the recombinant microorganism contains one or more or all of the following characteristics: (1) adding nicotinamide into a fermentation culture medium, and transforming by the recombinant microorganism to generate NMN; and (2) the nicotinamide phosphoribosyltransferase is overexpressed. And (3) deletion or inactivation or enzyme activity reduction of genes for encoding alkaline phosphatase and nucleotidase on the recombinant microorganism genome.

Bertrand(1982) hydrolytic generates trophicardyl from trophicardyl acid by nucleotidase, and gains H2O2 through the reaction of the enzyme1 and enzyme2. Use of H2O2 from Trinder's reaction to combine hydrogen provider3,5-double CL-2-hydroxide benzene sulphur acid and 4- to create a amaranth in the situation of the catalase. By watching the velocity of increase of the degree of the amaranth absorbing light at 510 nms to gain 5' nucleotidase activity. The invention uses the aniline compound ADOS, ADPS, ALPS, TODB, TOOS and TOPS as hydrogen provider for Trinder's reaction and increases the reaction ability. The invention also involves a reagent box used for implementing the method mentioned above.

The invention relates to a method for reducing T cell depletion and a medicine which has the function and can be used for clinical treatment. The medicament specifically comprises an anti-CD39 antibody medicament. According to the therapeutic dose level, the anti-CD39 antibody drug can reduce the expression quantity of CD39 on the surfaces of CD45 positive immune cells. And the CD39 structural domain recognized by the anti-CD39 antibody drug is included in the CD39 structural domain combined with the group of antibodies PB1, PB2, PB3, PB4 and PB5. The anti-CD39 antibody drug can also be effectively combined to Fc gamma RIIIa of cells.

Solid forms of Compound I, which modulates the conversion of AMP to adenosine by 5′-nucleotidase, ecto, and compositions containing the compound and methods for preparing the solid forms, are described herein. The use of such solid form of Compound I and compositions for the treatment and / or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, that are mediated by 5′-nucleotidase, ecto is also provided.

The invention discloses a liquid stable 5 '-nucleotidase calibrator. The calibrator comprises the following components: a buffer solution, a stabilizer, a preservative and 5'-nucleotidase. The invention also discloses a 5 '-nucleotidase detection kit. The kit comprises a 5'-nucleotidase R1 reagent and a 5 '-nucleotidase R2 reagent. The 5 '-nucleotidase R1 reagent is an enzyme reaction system and consists of a first buffer solution, a stabilizer, a first preservative, 4-aminoantipyrine, an enzyme activator, purine nucleoside phosphorylase (PNP), xanthine oxidase (XOD) and peroxidase (POD); the 5 '-nucleotidase R2 reagent is a chromogenic system and is composed of a second buffer solution, a second preservative, inosine-5'-disodium phosphate, beta-sodium glycerophosphate and a Trinder chromogenic substrate. The invention also discloses an application of the 5 '-nucleotidase detection kit in determination of 5'-nucleotidase.

This disclosure relates to antibodies that bind an epitope present on CD73 expressed at the surface of cells, including tumor cells, and that inhibit the enzymatic (ecto-5′ nucleotidase) activity of the CD73 enzyme. Such agents can be used for the treatment of diseases such as cancers.

The invention relates to a method for determining the activity of 5'-nucleotidase, further to a 5'-nucleotidase detection kit, and belongs to the technical field of medical test determination. According to the method, 5-inosine monophosphate used as a substrate is converted and dehydrogenated by using purine nucleoside phosphorylase and xanthine dehydrogenase as tool enzymes, oxidized coenzyme isreduced to form a reduced coenzyme, and the enzyme reaction rate is calculated by detecting the generation amount of the reduced coenzyme at a wavelength of 340 nm. Compared to the method in the priorart, the method of the present invention has advantages of less reaction steps, less kinds of raw materials, strong alkaline-phosphatase resistance, stable reagent performance, wide linear range, high sensitivity and good accuracy.

The present invention provides a 5'-nucleotidase detection kit and a preparation method thereof, belongs to the field of in-vitro diagnostic reagents, and solves the low precision and stability problems of a 5'-nucleotidase detection kit in the prior art. The kit comprises a reagent R1 and a reagent R2. The present invention also provides the preparation method of the 5'-nucleotidase detection kit. The R1 reagent contains a compound stabilizer comprising TritonX-305 and AES, the compound stabilizer can improve the stability of the reagent, and also can improve the dispersion of the reagent, reduce the absorbance difference between two times of blank calibration, and improve the kit precision; A90 contained in the R1 can effectively prevent precipitation of magnesium phosphate, expensive 18-crown ether-6 is replaced, the stability of the reagent is improved, reagent blank absorbance change rate is reduced, and the cost is reduced greatly; by addition of glycine and EDTA. 2K into the reagent R2, detection sensitivity and precision of the kit can be improved, and by addition of the glycine, reagent blank absorbance is reduced.

The invention relates to the technical field of biology, in particular to a system and a method for expressing and purifying recombinant protein with 3'-nucleotidase as a tag and application of the recombinant protein. The system is characterized in by comprising a 3'-nucleotidase nucleotide sequence and a protein over-expression vector, a 3'-nucleotidase nucleotide sequence added with a protease recognition site, a protease nucleotide sequence matched with the protease recognition site, a lysis solution containing at least one of Li<+> and Ca<2+> ions, an eluent of a chelating agent containing at least one of Li<+> and Ca<2+> ions, and PAP (Prostatic Acid Phosphatase) agarose gel. The system disclosed by the invention is a simple, high-efficiency and economic system for purifying a target recombinant protein.

Methods of identifying compounds that modulate the conversion of AMP to adenosine by 5′-nucleotidase, ecto, and that possess particular pharmacokinetic characteristics are described herein. Methods of such compounds, and compositions comprising same, for the treatment and / or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, are also provided.

The invention relates generally to the field of sodium and lithium ion detection. In particular, the invention provides chimeric proteins, nucleic acids encoding chimeric proteins, methods and kits for assaying for sodium ions and for lithium ions in a sample, using inter alia, a 3′(2′),5′-bisphosphate nucleotidase.

Compounds that modulate the conversion of AMP to adenosine by 5′-nucleotidase, ecto, and compositions containing the compounds and methods for synthesizing the compounds, are described herein. The use of such compounds and compositions for the treatment and / or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, that are mediated by 5′-nucleotidase, ecto is also provided.

The invention provides a method for preparing nucleosides of nicotinic acid or derivatives thereof, nicotinic acid adenine dinucleotide and nicotinic acid mononucleotide, an enzyme composition and application. The method for preparing the nucleosides of the nicotinic acid or the derivatives thereof comprises the step of carrying out reaction by using 5'-nucleotidase and mononucleotide of the nicotinic acid or the derivatives thereof or salt of the mononucleotide as a substrate, wherein the amino acid sequence of the 5'-nucleotidase is shown as SEQ ID NO.1. Compared with a chemical synthesis method, the method is safer and more reliable, and is more environment-friendly because the use of an organic solvent can be avoided. Nicotinamide nucleoside can be more effectively produced so as to meet increasing demands.

The invention discloses a kit for determining 5'-ribonuclease. The kit is composed of a reagent R1 and a reagent R2 which are independent of each other. The reagent R1 is prepared from the following components: a buffer solution, a preservative, a protective agent, an anti-interference agent, a surfactant, a reaction enhancer, peroxidase, purine nucleoside phosphorylase, xanthine oxidase, a chromogen substance and water; and the reagent R2 is prepared from the following components: a buffer solution, a protective agent, a reaction enhancer, a preservative, disodium inosinate, 4-aminoantipyrineand water. The kit is high in analysis sensitivity, good in repeatability and high in anti-interference capability, the linearity can reach up to 400U / L, the detection of abnormal specimen results conforms to the actual concentration, and the accuracy of normal specimen test results is not affected.

The invention relates to the technical field of biology, in particular to a system and a method for expressing and purifying recombinant protein with 3'-nucleotidase as a tag and application of the recombinant protein. The system is characterized in by comprising a 3'-nucleotidase nucleotide sequence and a protein over-expression vector, a 3'-nucleotidase nucleotide sequence added with a protease recognition site, a protease nucleotide sequence matched with the protease recognition site, a lysis solution containing at least one of Li<+> and Ca<2+> ions, an eluent of a chelating agent containing at least one of Li<+> and Ca<2+> ions, and PAP (Prostatic Acid Phosphatase) agarose gel. The system disclosed by the invention is a simple, high-efficiency and economic system for purifying a target recombinant protein.

The invention is about a method of measuring the activation of 5í»-phosphonuclease, and it also concerns the reagent box of 5í»-phosphonucleasediagnosis. This invention belongs to the field of medical testing and measuring technology. The reagent box is consisted of buffer solution, carnine monophosphate, adenosine triphosphate, phosphoenolpyruvate, reduced coenzyme, carnine kinase, pyruvate kinase, lactate dehydrogenase and stabilizer. Firstly, we cause an enzyme-coupled reaction through mixing the sample and the reagent according to a certain proportion of volume; secondly, put the final reactant under the biochemical analyzer and test the absorbance variational situation (speed) of dominant wavelength; then we can get theactivation of 5í»-phosphonuclease. By using this invention, we can get the necessary measuring result with high sensitiveness and fine precision through biochemical analyzer, and the result would not be contaminated by material of internal and exogenous sources. Thus, this method can be conveniently promoted and applied.