39 results about "CYP1A2" patented technology

The invention discloses SNP rs762551 of a CYP1A2 gene and application thereof in relevant drug metabolism activity detection, and also provides a method for prejudging drug metabolism activity. The method comprises the steps of detecting whether the cytochrome oxidase P4501A2 gene (CYP1A2) and a transcript of a human individual have variation or not compared with a normal cytochrome oxidase P4501A2 gene and a normal transcript, and if so, showing that the drug metabolism activity of the individual is different from common or normal people, for the individual. The invention also discloses a corresponding detection kit used for prejudging the drug metabolism activity of the individual.

The invention relates to a preparation method, a detection method and application of a probe drug composition for determination of metabolic activity of cytochrome P450. The composition mainly comprises a preparation made with a specific probe with major isoforms of CYP450, i.e. CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A4, as an active component. Cocktail probe drug solution is prepared, the probe drug composition is injected into an animal or liver microsomes for in vitro co-incubation, and the concentration of each probe drug is determined to assess the metabolic activity of the CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A4. In the early stage of research and development of new drugs, the effects of the drugs on the activity of each isoform of the cytochrome P450 are screened in a high-throughput way, and the interactions of the drugs can be predicted. In the stage of clinical research, the testing can be performed with the probe drug composition in an in-vivo probe method, and the effects of the drugs on the in-vivo metabolic activity of different isoforms of the human liver CYP450 can be examined.

The invention relates to an absolute quantitative method for biomass spectra of CYP450 enzyme hypotypes, and belongs to the technical field of proteomics. In the method, on the basis of liquid chromatography (LC) / mass spectrometry (MS) / MS, the absolute quantification of four CYP450 enzyme hypotypes, namely CYP1A2, CYP2B6, CYP3A4 and CYP3A5 in systems such as liver microsome and the like is realized simultaneously by a strategy that CYP450 generates specific peptide quantitative enzymes by pancreatic enzyme hydrolysis. The method mainly comprises the following steps of: preprocessing a proteinsample, preparing a standard curve, measuring the protein sample and preparing a quality control sample. The absolute quantitative method has excellent linear relations, is high in accuracy and repeatability, is sensitive, precise and reliable, and can be used for medicinal metabolism and the evaluation of the mutual effect of medicaments.

The invention belongs to the technical field of molecular biological gene polymorphism detection, and in particular relates to a kit and a method for genotyping detection. The kit consists of a firstprimer pair and a first molecular beacon probe for ADH1B gene rs1229984 site, a second primer pair and a second molecular beacon probe for ALDH2 gene rs671 site, a third primer pair and a third molecular beacon probe for CYP1A2 gene rs762551 site, and a fourth primer pair and a fourth molecular beacon probe for TMPRSS6 gene rs855791 site. With the application of the kit provided by the invention,four genotypes corresponding to the genes and corresponding metabolic absorption types can be rapidly and conveniently detected.

The invention provides 2, 3, 5, 7-tetrasubstituted dihydro-pyrazolo piperidine derivative and a preparation method and application thereof. The derivative is 2, 3-bis(substituted phenyl)-5-subsituted arylmethyl-7-substituted benzylidene dihydro-pyrazolo piperidine derivative, having the following formula (I). The preparation method includes using substituted arylmethyl amine and methyl acrylate as raw materials; subjecting the materials to Michael addition, Dieckmann condensation and hydrolysis-decarboxylation sequentially; allowing for Aldol reaction with substituted benzaldehyde to obtain intermediate N-substituted arylmethyl-3, 5-bis(substituted benzylidene)-4-piperidone; allowing for condensation with substituted phenylhydrazine to obtain a compound according to the formula (I). The derivative is efficient in inhibiting multiplication of various carcinoma cell lines such as leukemia, esophagus cancer, ovarian cancer and breast cancer in human, is well stably metabolic in liver microsomes of human and rat, is free of direct and competitive inhibition on five enzymes of liver microsomes, such as CYP3A4, CYP2D6, CYP2C9, CYP1A2 and CYP2C19, is highly bioavailable, is low in toxicity to normal cells, and is available for the preparation of drugs for the cancers.

The invention discloses a method for predicting inhibition concentration of a cytochrome P450 enzyme CYP1A2 inhibitor by utilizing simplified partial least squares. The method comprises the steps of: 1) collecting, processing and optimizing sample sets; 2) constructing molecular descriptors of the inhibitor; 3) preliminarily screening molecular descriptors of the inhibitor; 4) scaling data sets of molecular descriptors of the inhibitor again; 5), dividing data sets of molecular descriptors of the inhibitor; 6), setting up a QSAR model; and 7) predicting inhibition concentration of the cytochrome P450 enzyme CYP1A2 inhibitor.The method utilizes the method of partial least squares (PLS) to further estimate and simplify constituents of partial least squares (PLS) in the presence of serious multiple correlations of independent variables on the basis of regression modeling and uses the simplified method of partial least squares (SIMPLS) to precisely predict inhibition concentration of the cytochrome P450 inhibitor.

The invention discloses a kit for screening colorectal cancer hereditary susceptibility genes. The kit comprises colorectal cancer susceptibility gene specificity primers for amplifying a plurality of target regions in a to-be-detected sample, wherein the colorectal cancer susceptibility genes include at least one of rs10795668, MMP2, SMAD7, ADH2, ALDH2, CYP1A2, MMP-1, MTHFR, TP53, VEGF, COX-2, DNMT3B, hMLH1, LOC727677, MMP9, MTRR and TGF-beta 1. The kit can detect a plurality of regions of the colorectal cancer susceptibility genes at the same time, detecting efficiency is improved, detecting cost is reduced, and detecting accuracy and sensitivity are high.

The invention discloses a liquid phase chip detection method and a detection kit for detecting the polymorphism of cytochrome P450 1A2 (CYP1A2) genes. The method comprises the following steps of: designing primers and probes for polymorphism CYP1A2*1F (-163C>A) and CYP1A2*1D (-2467delT) of the CYP1A2 genes, hybridizing a probe-microsphere mixture formed by covalent bonding of the probes and microspheres and a polymerase chain reaction (PCR) amplification product, adding streptavidin phycoerythrin, and thus detecting fluorescence signals of different microspheres and determining the polymorphism of the CYP1A2 genes in a sample to be detected. The method and the kit have the advantages of high sensitivity, high flux, quickness and accuracy in detection, and the like, can be used for detecting and identifying the polymorphism of the CYP1A2 genes, and also provide an important reference for medicament metabolism and personalized medication.

Disclosed is a method for determining whether a dog is an extensive metabolizer or a poor metabolizer in the rate of drug metabolism, by preparing a DNA sample from a dog, and determining a base corresponding to a base at position 1117 of a canine CYP1A2 gene (i.e., at position 87 of exon 4). According to the method, a CYP1A2 gene diagnosis of dogs (particularly beagles) used in a pharmacological effect test or a toxicity test can be rapidly carried out prior to the test, and thus the dogs can be easily divided into a group having a normal metabolic ability (an extensive metabolizer group) and a group having a low metabolic ability (a poor metabolizer group).

the invention discloses a gene CYP1A2 genotypic and detecting method and relative genotypic agent box and application, which provides molecular criterion for metabolic dose.

The invention relates to a method for detecting in-vivo CYP1A2 and CYP3A4 enzyme activity in earthworm through high-performance liquid chromatography-tandem mass spectrometry, and belongs to the technical field of the enzyme activity detection. The method comprises the following steps: (1) preparing earthworm microsome protein suspension; (2) adding the earthworm microsome protein suspension prepared in the step (1) in an incubation system containing two specific probe primers, warming to start enzymatic reaction, after the enzymatic reaction is terminated, detecting two specific metabolites produced after the enzymatic reaction is terminated by utilizing high-performance liquid chromatography-tandem mass spectrometry, and respectively computing CYP1A2 and CYP3A4 enzyme activity. The detection method disclosed by the invention is high in accuracy and precision and sensitivity, strong in stability, capable of measuring multiple CYP subenzyme activities in the earthworm body, thereby providing a detection method for exploring response mode on the soil pollutant by different CYP subenzyme of the earthworm.

The invention belongs to the technical field of genotyping and particularly relates to a primer, probe, kit and method for detecting subtypes of a human gene CYP1A2. The primer and the probe comprise the following nucleotide sequences: the nucleotide sequence of a forward primer is SEQ ID NO: 1: 5'-AAACTGAGATGATGTGTGGAGG-3'; the nucleotide sequence of a reverse primer is SEQ ID NO: 2: 5'-CACGCATCAGTGTTTATCAAA-3'; the nucleotide sequence of the probe is SEQ ID NO: 3: 5'-GTGGGCCCAGGACGCATGGTAGATGGA-3'. The kit comprises the primer, the probe, dNTPs, MgCl2, a mixed enzyme and positive and negative reference substances, wherein the mixed enzyme is prepared from a UNG enzyme and DNA polymerase with the activity of flap endonuclease. According to the method for detecting the subtypes of the gene CYP1A2 by using the kit, different subtypes of a target nucleic acid SNP locus can be detected by one fluorescent probe, so that the operating process is simplified, the cost is reduced, and the detection result is high in resolution and is rapid and accurate.

The teachings provided herein generally relate to compositions comprising rutaecarpine derivatives that activates CYP1A2 through enzyme induction. The uses for such a derivative can include removing caffeine from a subject, improving sleep, treating insomnia, treating caffeine toxicity, treating caffeine addiction and withdrawal symptoms, and the like. Caffeine is just one example of a substrate that can be removed using the derivatives taught herein, and other examples, including theophylline, are provided herein.

The invention discloses SNP rs11632814 of a CYP1A2 gene and application thereof in relevant drug metabolism activity detection, and also provides a method for prejudging drug metabolism activity. The method comprises the steps of detecting whether the cytochrome oxidase P4501A2 gene (CYP1A2) and a transcript of a human individual have variation or not compared with a normal cytochrome oxidase P4501A2 gene and a normal transcript, and if so, showing that the drug metabolism activity of the individual, for the individal, is different from common or normal peopl. The invention also discloses a corresponding detection kit used for prejudging the drug metabolism activity of the individual.

The invention relates to a novel cocktail probe medicament combination comprising caffeine, metoprolol, chlorzoxazone, tolbutamide and midazolam. The influence of hepatic ischemia preconditioning and hepatic ischemia-reperfu-sion injury on medicament metabolism activity of rat CYP1A2, CYP2C9, CYP2E1, CYP2D6, and CYP3A4 in vivo is evaluated through the change of pharmacokinetics parameters of the combination.

The invention discloses a N-substituted benzyl tetrahydropyridine with -5-substituted indole and preparation method and application thereof, and the structure is shown in the general formula (I): substituted benzene methylamine (ethylamine) and methyl acrylate are raw materials, and an intermediate N-substituted benzylpiperidine (phenylethylpiperidine)-4-ketone is obtained by three steps of reaction such as Michael addition, Dieckmann condensation and hydrolysis decarboxylation or the like in sequence, and the object (I) is obtained by condensation reaction of the intermediate and 5-substituted indole. The compound (I) can effectively inhibit proliferation of human leukemia K562, Jurkat, U937, THP-1 cell line, the human esophagus cancer ECA-109 cell line, human liver cancer SMMC-7721 cell line, human ovary cancer HO-8910 cell line, human breast cancer MCF-7 cell line, breast cancer MDA-MB-231 cell line; the compound has a good metabolism stability in the human and rat liver microsomes; The compound does not have mechanical inhibition effects for five enzymes of human liver microsomes such as CYP3A4, CYP 2D6, CYP2C9, CYP1A2 and CYP2C19 or the like; the compound can induce the cell cycle G2 / M retardance and promote cancer cell apoptosis and inhibit cancer cell propagation.

The present invention relates to the furanochalcone class of compounds of general formula A. The present invention particularly relates to the synthesis of furanochalcones and their CYP1A1, CYP1A2 and CYP1B1 inhibitory activity. In addition, the invention relates to the prevention or treatment of cancer caused by polyaromatic hydrocarbons (PAHs), 4-nitroquinoline-1-oxide, and N-nitroso-N-methylurea, heterocyclic amines, estrogen and 17β-estradiol, resulting from the inhibition of CYP1A1, CYP1A2 and CYP1B1 enzymes.

The invention discloses a gene polymorphism detection kit for guiding psychiatric medication. The kit comprises a specific primer group and a fluorescent probe group, wherein the specific primer group and the fluorescent probe group are used for detecting the rs1065852 site of a CYP2D6 gene, the rs1414334 site of an HTR2C gene, the rs1800497 site of an ANKK1 gene, the rs762551 site of a CYP1A2 gene, the rs489693 site of an MC4R gene and the rs1799978 site of a DRD2 gene. According to the invention, the primer group and the fluorescent probe group are utilized, and the template DNA is specifically amplified by using a fluorescent PCR amplification technology, so that the polymorphism of a plurality of sites of a plurality of genes related to psychotropic drugs can be accurately detected at the same time; ARMS primer design is adopted, mismatched bases are introduced, the specificity of primer amplification is improved, the sensitivity is high, and accurate genotyping can be conducted on human template DNA at the lowest DNA initial amount of 10 ng; the detection speed is high and the efficiency is high: the PCR reaction only needs less than 1 hour to complete the amplification reaction, the cost is lower, the required reagent consumables are all clinically common reagents, the cost is lower, and the clinical popularization is convenient.

The invention discloses SNP rs4646418 of a CYP1A2 gene and application thereof in relevant drug metabolism activity detection, and also provides a method for prejudging drug metabolism activity. The method comprises the steps of detecting whether the cytochrome oxidase P4501A2 gene (CYP1A2) and a transcript of a human individual have variation or not compared with a normal cytochrome oxidase P4501A2 gene and a normal transcript, and if so, showing that the drug metabolism activity of the individual is different from common or normal people, for the individual. The invention also discloses a corresponding detection kit used for prejudging the drug metabolism activity of the individual.

The invention belongs to the field of gene diagnosis, and discloses a real-time fluorescence PCR method of detecting a rs762551 site of a CYP1A2 gene, and a primer-and-probe combination of the real-time fluorescence PCR method. A sequence of one forward primer FpG is 5'-GGTGAGCTCTGTGGGGC-3', a sequence of the other forward primer FpG is 5'-GGTGAGCTCTGTGGGGA-3', a sequence of a reverse primer Rp is5'-GCTGAGGGTTGAGATGGAGAC-3', and a sequence of a probe Probe is 5'-Cy5-ACGCATGGTAGATGGAGCTTAG-BHQ2-3'. After a PCR is completed, results can be analyzed through whether or not an amplification curveexists and in combination with a Ct value. The real-time fluorescence PCR method of detecting the rs762551 site of the CYP1A2 gene is applied to polymorphism detection of the rs762551 site, and has the advantages of being easy to operate, high in resolution, free of pollution, and high in throughput.

The invention provides construction of a model of co-expression uptake of a carrier and a drug-metabolizing enzyme. The construction comprises the following steps: amplifying a human gene OAT1cDNA used as a template to obtain a human OAT1 gene segment, connecting the obtained human OAT1 gene segment to a carrier pcDNA3.1(+) to obtain a plasmid pcDNA3.1(+) / OAT1, transfecting MDCK (Madin Darby Canine Kidney) cells, and screening to obtain MDCK-hOAT1 cells in which the OAT1 is highly expressed; connecting a human CYP1A2 gene segment to a carrier pcDNA3.1(+) / Hygro(+) to obtain a plasmid pcDNA3.1 / Hygro(+) / CYP1A2, transfecting the MDCK-hOAT1 cells, screening by using a hygromycin B to obtain cells of co-expression of OAT1 and CYP1A2, then carrying out mRNA verification on the cells and also carrying out functional verification by virtue of a classic substrate and an inhibitor. The cell model can be applied to cellular-level screening based on the medical toxic effects of OAT1 and CYP1A2, and therefore, prediction of the synergistic effect of the two proteins in a medicine disposition process is realized and a basis can be provided for clinical rational medications. The cell model is rational in design and high in repeatability.

The invention provides novel application of a qi-tonifying and heart-rate-restoring preparation for injection and especially to application of the qi-tonifying and heart-rate-restoring preparation for injection in preparing medicines exerting induction action on the activity of the liver microsomes, CYP1A2 and CYP3A4.

the invention discloses a gene CYP1A2 genotypic and detecting method and relative genotypic agent box and application, which provides molecular criterion for metabolic dose.

The present invention relates to a preparation method of CYP1A2 polypeptide and antihuman CYP1A2 polypeptide antibody. The amino acid sequence of CYP1A2 polypeptide is GRERRPRLSDRPQLP. The preparation method of antihuman CYP1A2 polypeptid antibody includes the following steps: (1) analysis of CYP1A2 antigen epitope; (2) synthesis of CYP1A2 polypeptide; (3) CYP1A2 polypeptide and carrier protein crosslinking; (4) preparing rabbit anti-CYP1A2 polypeptide antibody; and (5) collecting and separating to obtain serum containing antibody, purifying antibody so as to obtain anti-human CYP1A2 polypeptide antibody.

The invention belongs to the technical field of biological medicine, and discloses a construction method of a CYP enzyme humanized rat animal model for drug metabolism research and application of the CYP enzyme humanized rat animal model. The construction method comprises genotype identification, humanized gene expression identification and humanized gene function verification. The method comprises the following steps of firstly, constructing a CYP enzyme humanized rat by utilizing a CRISPR system, including selection of a gene editing target spot, in-vitro synthesis and transcription of sgRNA and Cas9 mRNA, construction of a homologous recombination template, preparation of a pseudopregnant female rat, in-vitro microinjection and transplantation of a single-cell embryo and reproduction of the rat, finally obtaining a homozygote CYP gene humanized rat, then performing CYP enzyme expression detection on the homozygous humanized rat from a protein level, and performing metabolic function verification to prove that the CYP humanized rat is successfully constructed. The CYP1A2 humanized rat has metabolic characteristics more similar to those of people, the species difference of CYP1A2 mediated drug metabolism between the rat and the people is eliminated, and a new rat animal model is provided for research on CYP mediated drug metabolism.

The invention discloses an application of a reagent kit in accordance with ASMT / CYP1A2 molecules to forecasting clinical prognosis and immunity characteristics of solid tumors. The inventor performs bioinformation analysis of large samples through researching transcriptomes and genome data of 6658 tumor samples in all of 14 kinds of entity tumors, the relationship of the ratio level of ASMT / CYP1A2and clinical prognosis, tumor mutation level and tumor neoantigen load of patients in different entity tumors is confirmed, wherein the clinical prognosis of the patients suffering from breast cancerand gastric cancer in a High Index group is more considerable; the multifactor analysis result shows that the result of ASMT / CYP1A2 is an independent forecasting index of the clinical prognosis of the tumor patients; in addition, in patients suffering from bladder cancer, breast cancer and prostatic cancer, the prognosis of patients in a Low Index group is poor, but more mutational load and / or tumor neoantigen exists, which prompts that the patients in the Low Index group can obtain benefits from immunotherapy possibly.

The present invention discloses a gene polymorphism detection primer set, a kit and a detection method of a caffeine metabolic capacity related gene CYP1A2. The gene detection primer set comprises three detection primers and two internal control primers. The present invention provides the primer composition for detecting CYP1A2 gene mutations of samples of fresh blood, etc., and when the primer composition is used for detecting a C5246A mutation of the CYP1A2 gene, the primer composition has advantages of high sensitivity, strong specificity, short time consumption, etc. The kit is simple instructure design and convenient to use. The partition design enables the detection process to be less susceptible to contamination and detection results are more accurate. The detection method is short in time consumption, high in sensitivity and strong in specificity.

The invention relates to an absolute quantitative method for biomass spectra of CYP450 enzyme hypotypes, and belongs to the technical field of proteomics. In the method, on the basis of liquid chromatography (LC) / mass spectrometry (MS) / MS, the absolute quantification of four CYP450 enzyme hypotypes, namely CYP1A2, CYP2B6, CYP3A4 and CYP3A5 in systems such as liver microsome and the like is realized simultaneously by a strategy that CYP450 generates specific peptide quantitative enzymes by pancreatic enzyme hydrolysis. The method mainly comprises the following steps of: preprocessing a proteinsample, preparing a standard curve, measuring the protein sample and preparing a quality control sample. The absolute quantitative method has excellent linear relations, is high in accuracy and repeatability, is sensitive, precise and reliable, and can be used for medicinal metabolism and the evaluation of the mutual effect of medicaments.

The invention relates to a novel cocktail probe medicament combination comprising caffeine, metoprolol, chlorzoxazone, tolbutamide and midazolam. The influence of hepatic ischemia preconditioning and hepatic ischemia-reperfu-sion injury on medicament metabolism activity of rat CYP1A2, CYP2C9, CYP2E1, CYP2D6, and CYP3A4 in vivo is evaluated through the change of pharmacokinetics parameters of the combination.

The invention discloses a method for predicting inhibition concentration of a cytochrome P450 enzyme CYP1A2 inhibitor by utilizing simplified partial least squares. The method comprises the steps of: 1) collecting, processing and optimizing sample sets; 2) constructing molecular descriptors of the inhibitor; 3) preliminarily screening molecular descriptors of the inhibitor; 4) scaling data sets of molecular descriptors of the inhibitor again; 5), dividing data sets of molecular descriptors of the inhibitor; 6), setting up a QSAR model; and 7) predicting inhibition concentration of the cytochrome P450 enzyme CYP1A2 inhibitor.The method utilizes the method of partial least squares (PLS) to further estimate and simplify constituents of partial least squares (PLS) in the presence of serious multiple correlations of independent variables on the basis of regression modeling and uses the simplified method of partial least squares (SIMPLS) to precisely predict inhibition concentration of the cytochrome P450 inhibitor.