72 results about "Nicotinamide phosphoribosyltransferase" patented technology

The invention discloses a novel method for preparing beta-nicotinamide mononucleotide (beta-NMN). According to the method, D-5-ribose phosphate, ATP and nicotinamide are used as raw materials, beta-nicotinamide mononucleotide can be efficiently and biologically synthesized by catalysis of immobilized whole-cell containing phosphoribosylpyrophosphate synthetase and nicotinamide phosphoribosyltransferase, and the concentration of the synthesized beta-nicotinamide mononucleotide can be 13.3g / L, so that the synthetic amount and conversion rate of beta-NMN enzymatic reaction can be increased, the immobilized whole-cell can be used repeatedly, and the reaction complexity and production cost can be reduced.

The invention relates to methods of treating diseases, particularly cancers, that respond favorably to the inhibition of Nicotinamide phosphoribosyltransferase (Nampt); it also relates to therapeutic methods that utilize Nampt inhibitors in combination with NAD biosynthesis precursors to intentionally kill cancer cells while limiting or minimizing toxicity to normal host cells; and it relates to methods of identifying cancers that will be most responsive to treatment with Nampt inhibitors, particularly when administered in combination with nicotinic acid.

Crystals of nicotinamide phosphoribosyltransferase, methods of making the crystals, and methods of using the crystals are disclosed. The three-dimensional structures of NMPRTases are also disclosed. Also disclosed are methods for utilizing a crystal structure of an NMPRTase for identifying, designing, selecting, or testing molecules which affect NMPRTase activity, which can be used therapeutically in the treatment of diseases and disorders such as cancer and diabetes.

The invention relates to the technical field of biological science and technology and provides a nicotinamide phosphoribosyltransferase mutant, a recombinant expression vector and a recombinant bacterium containing the mutant, and application in beta-nicotinamide mononucleotide synthesis. In the nicotinamide phosphoribosyltransferase mutant, 1 or 2 amino acid residues in amino acids selected fromamino acid equivalent positions of sites 156, 174, 203, 231, 242, 258, 272 and 405 in an amino acid sequence of a wild-type nicotinamide phosphoribosyltransferase are replaced by other amino acid residues. The nicotinamide phosphoribosyltransferase mutant disclosed by the invention increases the enzyme activity of the nicotinamide phosphoribosyltransferase, can promote synthesis of a beta-nicotinamide mononucleotide and can promote production of the beta-nicotinamide mononucleotide.

The present invention provides a method for preparing nicotinamide mononucleotide (NMN) by biocatalysis. The method includes a step of catalytically reacting a plurality of raw materials including nicotinamide, ATP, and ribose in the presence of nicotinamide phosphoribosyltransferase (Nampt), ribose phosphate pyrophosphokinase, and ribokinase, to prepare the NMN.

The invention provides a nicotinamide ribosyltransferase mutant and application thereof. Compared with an amino acid sequence SEQID NO. 2, the difference of the amino acid sequence of the mutant is that the R189th, S232th and R302th sites in the amino acid sequence SEQID NO. 2 are subjected to single mutation or in-pair combined mutation or three combined mutation. Novel nicotinamide ribosyltransferase mutant enzyme is used for synthesis and preparation of beta-nicotinamide mononucleotide. The constructed nicotinamide ribosyltransferase mutant enzyme has the advantages that the enzyme cost islow, the transformation time is short, and the process operation is simple, and has a broad large-scale industrial application prospect.

The present application discloses compounds of formula (I) wherein X is ═O, ═S, ═NH, ═NOH and ═NO-Me; A is —C(═O)—, —S(═O)2—, —C(═S)— and P(═O)(R5)—; B is, —O—, —(CH2)3-6—, and O—(CH2)2-5—; D is, —O—, —CR7R8— and —NR9; m is 0-12, n is 0-12, m+n is 1-20; p is 0-4; R1 is opt.sub. heteroaryl; and pharmaceutically acceptable salts thereof, and prodrugs thereof. The application also discloses the compound for use as a medicament for the treatment of a disease or a condition caused by an elevated level of nicotinamide phosphoribosyltransferase (NAMPRT), e.g. inflammatory and tissue repair disorders; dermatosis; autoimmune diseases, Alzheimers disease, stroke, athersclerosis, restenosis, diabetes, glomerulonephritis, cancer, cachexia, inflammation associated with infection and certain viral infections, including Acquired Immune Deficiency Syndrome (AIDS), adult respiratory distress syndrome, ataxia telengiectasia.

The invention provides compositions for the diagnosis or treatment of neoplasias, including lymphomas, leukemias, brain cancers (e. glioblastomas, medulloblastomas), breast cancer, colon cancer, and pancreatic cancer, and methods of use therefor.

The present invention discloses a Nicotinamide phosphoribosyltransferase (nampt) mutant and use thereof. The present invention relates to a nicotinamide phosphoribosyltransferase (Nampt) mutant artificially obtained through genic site-directed mutation. An object of the present invention is to provide a Nampt mutant having a catalytic activity higher than that of a conventional wild type parent, wherein the enzymatic activity of the Nampt mutant provided in the present invention is 1.2-6.9 times of the enzymatic activity of the parent.

Belonging to the pharmaceutical field, the invention relates to application of nicotinamide phosphoribosyltransferase (NAMPT) in preparation of neuroprotective and rehabilitation drugs. Experiments of the invention show that NAMPT recombinant protein can penetrate the blood-brain barrier to significantly reduce the volume of ischemic brain injury and alleviate white matter injury after ischemic brain injury; the NAMPT recombinant protein can significantly improve long-term behaviors after ischemic brain injury; the NAMPT recombinant protein can raise revascularization related factors to enhance revascularization after brain hypoxia and neuron regeneration after brain hypoxia, and promote post-injury neurologic function reestablishment; and by mediating oligodendrocyte regeneration and alleviating the demyelination effect of local lysolecithin, the NAMPT recombinant protein can promote restoration of white matter after injury and strengthen post-injury neurologic function reestablishment. Results show that the NAMPT recombinant protein plays a long-term neuroprotective role in drugs treating cerebral stroke, traumatic brain injury and multiple sclerosis, and can be used as a neuroprotective drug to treat acute and chronic injury caused by brain stroke.

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.

The invention discloses an enzyme gene suitable for efficiently synthesizing an NAD (Nicotinamide Adenine Dinucleotide) derivative in microorganisms, and belongs to the technical field of gene engineering and bioengineering. According to the invention, nicotinamide phosphoribosyltransferase gene VpNadV shown as any one of SEQ ID NO.1-7 is expressed in escherichia coli, and a pncC gene, a ushA gene, an nadR gene and a purR gene are knocked out, so that the accumulation amount of the NAD derivative in escherichia coli cells is increased, and fermentation conditions are optimized, so that the accumulation amount of the NAD derivative is further increased, the NAD derivative with the concentration of 1g / L or above can be obtained under a 5L fermentation system, and the enzyme gene has wide application prospects in the fields of foods, medicines, cosmetics, feeds and textiles.

A method of making nicotinamide mononucleotide (NMN), nicotinamide mononucleotide derivatives, or mixtures thereof is disclosed. The method involves the in vitro artificial enzymatic pathways comprised: the generation of alpha-D-ribose-1-phosphate from numerous substrates followed by the synthesis of nicotinamide mononucleotide catalyzed by nicotinamide riboside phosphorylase and nicotinamide riboside kinase or the generation of 5-phospho-alpha-D-ribose-1-diphosphate from nucleotides followed by the synthesis of nicotinamide mononucleotide catalyzed by nicotinamide phosphoribosyltransferase. The multiple enzymes were reconstituted in one pot, wherein in-situ removal of byproducts that can be converted to other non-inhibitory chemicals with supplementary enzymes push the overall biotransformation toward the synthesis of nicotinamide mononucleotide. Furthermore, nicotinamide mononucleotide can be converted to its derivatives—nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate.

The present application discloses a compound of the formula (I) wherein Q is optionally substituted pyridyl; p is 0-6. Y is formulae (i), (ii) and (iii) where X is ═O, ═S and ═N—CN, r is 1-12, R is —Z-A, Z is a single bond, —S(═O)2—, >P═O, >C═O, —C(═O)NH—, and —C(═S)NH—; and A is hydrogen, C1-12-alkyl, C3-12-cycloalkyl, —[CH2CH2O]1-10—(C1-6-alkyl), C1-12-alkenyl, aryl, heterocyclyl, and heteroaryl; B is a single bond, —NRN—, —S(═O)2— and —O—; wherein RN is selected from hydrogen, C1-12-alkyl, C3-12-cycloalkyl, —[CH2CH2O]1-10—(C1-6-alkyl), C1-12-alkenyl, aryl, heterocyclyl, and heteroaryl; s is 0-6; and Cy is aryl, cycloalkyl, heterocyclyl, and heteroaryl. The compounds are useful for use as a medicament for the treatment of a disease or a condition caused by an elevated level of nicotinamide phosphoribosyltransferase (NAMPRT).

The invention relates to a combined drug, comprising a certain amount of nicotinamide phosphoribosyltransferase (NAMPT) depressor and a certain amount of NQO1 substrate. The NAMPT depressor and the NQO1 substrate have synergistic effects in the anti-tumor process, and the action of the composition in the case of combined utilization of the NAMPT depressor and the NQO1 substrate is superior to the superposition when all the drugs are independently used. The combined drug can be applied to treatment of the non-small cell lung cancer and other cancers. Therefore, a novel treatment method for treating the non-small cell lung cancer is provided by the invention.

The invention belongs to the field of chemical medicine, and discloses a benzo nitrogen hetero-aromatic ring compound represented by formula I, or pharmaceutically acceptable salts, stereisomers, racemic compounds, prodrugs, or solvates thereof. The invention also discloses applications of the benzo nitrogen hetero-aromatic ring compound in preparation of drugs used for treating diseases caused byprotein kinase and / or nicotinamide phosphoribosyltransferase abnormal activity. The benzo nitrogen hetero-aromatic ring compound represented by formula I or the salts thereof possess tyrosine kinaseand Nampt double inhibition effects, can be taken as effective components in treatment or prevention of tumor, are excellent in curative effect, and low in toxic or side effect.

The present application discloses a method for the treatment or for alleviating the symptoms of a cancer in a subject comprising the steps of a) determining the level of Nicotinic acid phosphoribosyltransferase (NAPRT) in said subject; and b) 1) in the event of a level of NAPRT which is lower than a predetermined threshold value, treating said subject sequentially / simultaneous with i) an effective amount of a nicotinamide phosphoribosyltransferase inhibitor (NAMPRTi), and ii) an effective amount of a nicotinic acid, a nicotinic acid precursor or a prodrug of nicotinic acid; or 2) in the event of a level of NAPRT which is higher than or equal to a predetermined threshold value, treating said subject with i) an effective amount of a NAMPRTi in the absence of sequential / simultaneous treatment with ii) an effective amount of a nicotinic acid, a nicotinic acid precursor or a prodrug of nicotinic acid.

The invention belongs to the field of biological medicine detection, and particularly relates to a biomarker for auxiliary diagnosis of gastric cancer and application of the biomarker, nicotinamide phosphoribosyltransferase is adopted as the marker for auxiliary diagnosis of gastric cancer and is used for preparing a diagnostic reagent or kit for detecting gastric cancer. Wherein a sample used for detecting the gastric cancer by the diagnostic reagent or the kit is serum, plasma or body fluid. The biomarker has the advantages that the drawn ROC curve shows that compared with CEA and CA19-9, the NAMPT has better specificity and sensitivity, and an experimental basis is provided for taking the NAMPT as a gastric cancer early-stage risk prediction index. And the positive rate of the three combined auxiliary diagnosis of the gastric cancer is 90%. The level of NAMPT in serum of a gastric cancer patient before an operation is detected by adopting an enzyme-linked immunosorbent assay, and retrospective analysis finds that the survival rate of the gastric cancer patient with the high NAMPT level before the operation is remarkably poor. This indicates a potential marker for poor prognosis of high NAMPT gastric cancer.

The invention discloses a method for preparing crosslinked NAMPT (Nicotinamide Phosphoribosyltransferase) and screening a NAMPT inhibitor and relates to the field of inhibitors. The method comprises the following steps: preparing a NAMPT recombinant protein; reducing the NAMPT recombinant protein with a reducing agent; carrying out a reaction between the reduced protein and a crosslinking agent so as to obtain the crosslinked NAMPT the active center of which is plugged; compounding a first protein solution and a second protein solution; preparing a first solution to be analyzed, a first contrast solution, a second solution to be analyzed and a second contrast solution; respectively measuring the fluorescence intensities of the first solution to be analyzed, the first contrast solution, the second solution to be analyzed and the second contrast solution by using a fluorescence spectrophotometer; and calculating the descending percentage of the fluorescence intensity of the first solution to be analyzed and the second solution to be analyzed. The method comprises few steps, is simple to operate and low in cost; and the external environment hardly influences a reaction process. Thus, the method is good in repeatability and the result is relatively accurate.

The invention discloses 4,5-dihydroisoxazole derivatives as NAMPT inhibitors. The present invention provides 4,5-dihydroisoxazole derivatives of formula (I) (as shown in the description), which may be therapeutically useful, more particularly as NAMPT inhibitors, wherein, ring A, L1, L2, X1, X2, X3, Z, R1, R2, R3, R4, R5, m, n, p and q have the meanings given in the specification and pharmaceutically acceptable salts thereof that are useful in the treatment and prevention of diseases or disorder, caused by an elevated level of nicotinamide phosphoribosyltransferase (NAMPT) in a mammal. The present invention also provides preparation of the compounds and pharmaceutical formulations comprising at least one of the substituted 4,5-dihydroisoxazole derivatives of formula (I) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

The invention relates to application of a diagnosis marker for diagnosing the breast cancer. Nicotinamide phosphoribosyltransferase (NAMPT), vascular endothelial growth factors (VEGF) and human epidermal growth factor receptor-2 (HER2) are combined to serve as the diagnosis marker. The positive rate of NAMPT in single immunohistochemical pathological diagnosis on the breast cancer is 54%, the positive rate of VEGF in single immunohistochemical pathological diagnosis on the breast cancer is 65%, and the positive rate of HER2 in single immunohistochemical pathological diagnosis on the breast cancer is 60%. The positive rates are low in single diagnosis on the breast cancer for the NAMPT, VEGF and HER2; the positive rate of diagnosis on the breast cancer through combination of the NAMPT, VEGF and HER2 is 90%. An enzyme linked immunosorbent assay (ELISA) is adopted for detecting expression conditions in serum before and after an operation performed on a patient with the breast cancer, and the detection result shows that NAMPT, VEGF and HER2 before the operation performed on the patient with the breast cancer are 6.64 times, 1.76 times and 2.52 times that of a health control group respectively (p value is smaller than 0.001), and after the operation, NAMPT, VEGF and HER2 are all decreased.

The present disclosure is directed to methods (e.g., in vitro methods) for use of nicotinamide phosphoribosyltransferase (NAMPT) as a biomarker in radiation-induced lung injury (RILI). Provided herein is an in vitro method for the diagnosis, prognosis, and / or monitoring of RILI in a human subject by providing a tissue or plasma sample from the subject and detecting the level of NAMPT therein, wherein a higher level of NAMPT in the tissue or plasma sample from the subject compared to a healthy control or a reference value is indicative for the presence of RILI in the subject. Further provided herein is a method of detecting NAMPT in a human subject by obtaining a biological sample from the subject, detecting the presence of NAMPT in the sample by contacting the sample with a capture agent that specifically binds NAMPT, and detecting binding between NAMPT and the capture agent.

The invention discloses an extraction and purification method of nicotinamide ribose phosphate transferase, which comprises the following steps: (1) carrying out fermentation culture on an NAMPT production strain to obtain fermentation liquor, centrifuging, and collecting thalli in the fermentation liquor; (2) adding pure water into the thalli, adjusting the pH value to 7.2-7.4 by using ammonia water, passing through a homogenizer, centrifuging, and separating to obtain supernate; (3) sequentially carrying out activated carbon decoloration and ceramic membrane filtration treatment on the supernate to obtain filtrate; (4) adding a nonionic detergent into the filtrate to remove inclusion bodies so as to obtain crude enzyme liquid; and (5) carrying out affinity chromatography on the crude enzyme liquid by adopting a nickel column, collecting eluent, carrying out electrodialysis desalination on the eluent, concentrating, crystallizing and drying, and extracting and purifying to obtain the NAMPT. By adopting the method disclosed by the invention, the NAMPT can be effectively extracted and purified from the fermentation liquor containing the nicotinamide phosphoribosyltransferase, and the yield and the purity of the NAMPT are remarkably improved.

The invention discloses a dioxobenzothiazole compound represented by a general formula I (shown in the description) or a pharmaceutically acceptable salt, a preparation method of the benzothiazole quinone compound and application of the benzothiazole quinone compound in the preparation of anti-tumor drugs. The structure of the compound is represented by a formula (shown in the description), wherein n represents 2-8, and X is selected from -CH2-, -NH-, -O- or formula; and R is selected from formula shown in the description. Compared with the prior art, a mother nucleus structure of a natural product pronqodine A is spliced with a key pharmacophore of an NAMPT inhibitor by virtue of a splicing principle so as to form a molecule. The compounds have very good anti-tumor activities in moleculeand cell levels, are capable of generating a large number amount of active oxygen substances in high-expression NQO1 tumor cells and effectively inhibiting the activity of nicotinamide phosphoribosyltransferase (NAMPT) and can be used for preparing anti-tumor drugs.