31 results about "In vitro metabolism" patented technology

The invention discloses a probiotic fermentation calcium supplement. The probiotic fermentation calcium supplement is obtained by selecting soybeans, germinating, grinding with a colloid mill, adding a certain amount of water, carrying out high pressure extraction and enzymolysis treatment, adding glucose, peptone and calcium, sterilizing, then fermenting by virtue of multiple probiotics, centrifuging and taking centrifugal liquid, adding a sweetening agent and prebiotics and sterilizing. The probiotic fermentation calcium supplement has the advantages that the probiotics are utilized for carrying out in vitro metabolism, secondary metabolites enter intestinal tracts to be directly absorbed by human body to take effect, negative effects brought about as thalli can not be colonized are avoided, and lactobacilli in the probiotics are utilized for natural metabolism on inorganic calcium to produce organic calcium lactate, so that calcium can be absorbed by the human body more easily, and the probiotic fermentation calcium supplement is a safe, green, environment-friendly, organic and healthy calcium supplement.

The invention relates to an LC-MC method used for determining NNK metabolites in liver microsomes and specifically to an LC-MC method used for analyzing and determining 4-(methyl nitroso)-1-(3-pyridyl)-1-butanone (NNK) metabolites in liver microsomes, which belongs to the field of biochemical analysis. The method provided in the invention mainly comprises the following steps: (1) preparation of an incubation system for liver microsomes; (2) sample treatment; (3) sample analysis and determination of NNK metabolites in the liver microsomes by using the LC-MC method. The invention has the following advantages of simple pre-treatment, a fast analysis speed, good selectivity, high detection sensitivity and capacity of analyzing and comparing the metabolism of NNK in different kinds of liver microsomes, can lay a methodological foundation for further research on in-vivo and in-vitro metabolism of NNK and other nitrosamine compounds and has critical significance.

The invention relates to a culturing method for body fluid derived tumor cells and a tumor medium. The method comprises the steps of enriching body fluid tumor cells; mixing the body fluid tumor cellswith an extracellular matrix; and in the presence of a basic medium, adding corresponding pleural effusion, ascites or cerebrospinal fluid and the like to carry out 3D cell culture. By the method, apatient derived tumor analog (PDTA) can be obtained effectively, and a novel detecting tool is provided for detection of sensitivity of clinical tumor patients to drugs and measurement of in-vitro metabolic stability, metabolic profiling, toxicity and the like of drug candidates.

The invention belongs to the technical field of genetic engineering and particularly discloses a soybean low-phosphorus-resistance related gene GmACP2, encoded protein and application of the GmACP2. The nucleotide sequence of the gene is shown by the Seq ID NO. 1. The expression quantity of the gene GmACP2 in the low-phosphorus-resistance material is remarkably higher than that of the low phosphorus sensitive material; the carrier which can guide expression of an exogenous gene in the plant is used, the gene GmACP2 is guided into the soybean hairy root, and the soybean phosphorus content, the dry matter accumulation and the phosphorus absorption and utilization ratio can be remarkably improved. The disclosed gene can be guided into the plant as a target gene, the phosphorus in-vitro metabolism balance capability of a transgenic plant is improved, and great significance is provided for cultivating soybean varieties with efficient phosphate use.

The invention discloses naphthoquinone compound alkannin, which can be used for quantitatively testing the enzyme activity of glucuronic acid transferase UGT1A9 and UGT1A8 as UGT1A9 and UGT1A8 specific probe substrate. The use is implemented by the following step of: selecting alkannin as a specific probe substrate, carrying out the UGT catalytic reaction of a specific substrate by using a UGT in-vitro incubation system, quantitatively testing the enzyme activity of UGT1A9 and UGT1A8 in biological samples and cells by quantitatively testing the lost amount of the substrate or the generated amount of the product. The substrate can be used for quantitative evaluation of the enzyme activity of UGT1A9 / 1A8 in different species, individuals and biological samples, calibration of propofol anesthetic metabolizing capacity of certain tissues, and calibration and estimation of the elimination rate of compounds which are mainly metabolized by UGT1A9 in specific organisms based on in-vitro metabolism dynamic data.

The invention provides application of a preclinical pharmacokinetic key technology and research system in cefoperazone sodium and sulbactam sodium, comprising the steps of (1) after the cefoperazone sodium and sulbactam sodium with a certain concentration is administrated to an experimental animal for a certain time, collecting one or more biological samples in blood, urine and faeces; (2) treating the biological samples obtained in step (1) by adopting liquid-liquid extraction, albumen precipitation, and solid phase extraction technologies to obtain corresponding solutions; (3) analyzing the prepared solutions in step (2) by adopting an LC-MS(liquid chromatography-mass spectrography) and an LC-MS / MS((liquid chromatography-tandem mass spectrometry). According to the application provided by the invention, the Caco-2 cell can also be adopted to detect the membrane permeability of the medicine and cell absorptive capacity; or the medicine with a certain concentration treats primary culture cerebral microvascular endothelial cells to detect the blood-brain barrier permeability of the medicine; or by adopting a whole animal, S9, human intestinal microsome and monoclonal purified enzyme, in vitro metabolism stability of the medicine is detected and a metabolite is identified.

The present invention provides a method for high throughput determination of the in vitro metabolism stability of a compound, and applications thereof. According to the present invention, the high-throughput problem is solved by using the 96-well plate; the reaction is initiated by adding NADPH in the reverse order while the termination reagent is added to terminate the reaction at the same time so as to solve the acetonitrile volatilization problem; and the control group with no NADPH addition is designed, such that the recovery rate can be calculated so as to find and screen the compounds independent of NADPH metabolism.

The invention discloses a method of quickly searching an in-vitro metabolite of a target compound and researching the subtype of a metabolic enzyme thereof by means of in-vitro human liver microsome incubation with combination of LC-Q-TOF / MS detection. The method is mainly used for researching of medicine-medicine interaction and mainly includes the steps of in-vitro incubating the target compound by the human liver microsomes, performing data collection and quick analysis to a sample by means of LC-Q-TOF / MS, and quickly searching the metabolites of the target compound and determining the subtype of the metabolic enzyme participating in the metabolism thereof. The method comprehensively considers the in-vitro human liver microsome incubation conditions and conditions of chromatography and mass spectrum data collection. In addition, with combination of related software for quickly analyzing the data, the metabolite of the target compound can be more accurately and quickly searched and found, and the subtype of the metabolic enzyme participating in the metabolism can be comprehensively determined, thus providing evidence for researching the medicine-medicine interaction.

The invention belongs to the technical field of medicine detection, discloses an LC-MS / MS (liquid chromatography-tandem mass spectrometry) method for determining cordycepin metabolite in liver microsome, and particularly relates to the LC-MS / MS method for analyzing and determining the cordycepin metabolite in the liver microsome. The LC-MS / MS method mainly includes the steps of (1) incubation of the liver microsome containing cordycepin; (2) preprocessing of a sample; (3) analysis of the sample, thereby determining the cordycepin metabolite in the liver microsome. The LC-MS / MS has the advantages of convenience in operation, sample processing simplicity, high analysis speed, high specificity and high flexibility, provides technical support for researching extrametabolites of the cordycepin in the liver microsome while laying a methodological foundation for researching in-vivo and in-vitro metabolites of the cordycepin, and accordingly has a great significance.

The invention discloses a shikonin oxime derivative containing an aza-side chain, a preparation method and medical application thereof. The structure of the derivative is shown in a formula (I), the formula (I) is shown in the description, wherein R represents a nitrogen-containing heterocyclic ring; the nitrogen-containing heterocyclic ring comprises a substituent-containing pyridine, pyrimidine,pyrazine, pyridazine, thiazole, benzazole, morpholine, piperidine, piperazine, N-methylene-N-hydroxyethyl piperazine, N-methylene-N-hydroxybutyl piperazine and a physiologically acceptable salt thereof; n is any positive integer ranging from 1 to 4. A compound disclosed by the invention is simple in preparation method, mild in reaction condition, and relatively high in yield. The prepared shikonin naphthazarin parent nucleus hydroxymethylated carbonyl oxime derivative containing the aza-side chain has the advantages of novel structure and good water solubility. Experimental studies on in-vitro metabolism and antitumor activity show that the compound plays a role through a prodrug mechanism and is relatively strong in antitumor activity.

The invention discloses flavonoid compounds which can treat neurodegenerative disease and have the structures represented by the following structure general formula I, II and III, and preparation methods and an application thereof. Physicochemical properties and in-vitro metabolism experiments prove that the compounds have better physicochemical and metabolic properties compared with scutellarin and seutellarein, and can be applied in preparation of drugs for treating the neurodegenerative disease.

The invention discloses lactobacillus rhamnosus capable of regulating and controlling relative abundance of acinetobacter in intestinal tracts, which belongs to the technical field of microorganisms and medicines. The lactobacillus rhamnosus CCFM1039 screened by the invention can be used for improving the intestinal health, which is specifically reflected in that (1) fructo-oligosaccharide, xylooligosaccharide and galactooligosaccharide can be utilized, and the lactobacillus rhamnosus CCFM1039 has a good oligosaccharide utilization capability; (2) various short-chain fatty acids such as aceticacid, propionic acid and butyric acid can be produced through in-vitro metabolism; and (3) the relative abundance of Acinetobacter in intestinal tracts can be reduced, so that the lactobacillus rhamnosus CCFM1039 has a huge application prospect in the preparation of products (such as foods, medicines or health care products) for improving the health of the intestinal tracts.

The invention discloses a method for detecting the metabolic performance of a new chemical entity in different species of liver microsomes, which comprises the following steps: in a reaction system containing a buffer solution, liver microsomes, a substrate, MgCl2 and NADPH, screening out an optimal reaction condition by taking the metabolic rate of the substrate as a reference to obtain a human liver microsome incubation system. According to the establishment method of the human liver microsome incubation system, rat, mouse, dog and monkey liver microsome incubation systems are sequentially established. The method comprises the following steps: performing in-vitro metabolism test on a new chemical entity in incubation systems of different species of liver microsomes, performing positive group reaction by using a substrate, and setting a negative group without adding NADPH and a blank group without adding liver microsomes; and identifying a metabolism result after testing. According to the method for detecting the metabolic performance of the new chemical entity in different species of liver microsomes, false positive and false negative results can be avoided, the reliability of an identification result is improved, and a reliable basis is provided for promoting research and development of new drugs.

The invention provides an in-vitro metabolic activity detection system based on intestinal microorganisms. The in-vitro metabolic activity detection system comprises the following steps of acquiring excrement samples, and screening the excrement samples through an excrement detector; according to a screening result, reserving excrement samples without organic lesions; carrying out activity detection on the excrement samples without the organic lesions; detecting the metabolic value of live intestinal bacteria according to an activity detection result; comparing the metabolic value of the intestinal bacteria with pre-stored information, and performing gene sequencing on the intestinal bacteria with large difference in the comparison value; determining structural characteristics of intestinal flora according to a gene sequencing result; and obtaining a biomarker at the early stage of diabetes according to the structural characteristics of intestinal flora. An intestinal microorganism in-vitro metabolic activity detection system is used for carrying out in-vitro fermentation on excrement of different subtypes and normal people, metabolic response values corresponding to different prebiotics are detected, including gas production, acid production, degradation degree of the prebiotics and promotion effect on probiotics, and therefore, the overall metabolic level of intestinal microorganisms is comprehensively reflected.

The invention discloses a novel method for researching the phase II metabolism of a flavone compound by using a model organism zebra fish. In the novel method, the zebra fish is selected as an object of drug metabolism research, experiment grouping design is carried out through an optimized experiment grouping method, drug administration is carried out by adopting an optimized drug administration mode, a metabolite of the flavone compound metabolized by the zebra fish is extracted by adopting an optimized method, the metabolite is analyzed by especially adopting an optimized analysis method, thus, the integral experimental method is high in operability and high in accuracy degree of an experimental result, the real conditions of the phase II metabolism of drugs in vivio and especially the real conditions of the phase II metabolism of the flavone compound in vivio can be objectively reflected, and the defect that a general in-vitro metabolism experiment is difficult to reflect a comprehensive result of in-vivo metabolism and the defects of large used drug quantity and high labor intensity of a general in-vivo metabolism experiment can be overcome. The experimental method has the advantages of high repeatability, small quantity of needed experimented drugs, low cost, low labor intensity, wide application range and high work efficiency by carrying out experiment research in a batched way.

The invention relates to a human liver microsome and cell co-culture system and a construction method and application thereof. Sterility of a human liver source is very difficult to ensure, and when human-derived liver microsome is prepared, the sterile liver microsome is hardly obtained. In order to carry out in-vitro metabolism test research on human-derived liver microsome and cell co-culture, the invention provides a co-culture method capable of enabling the germy human liver microsome to be in direct contact with sterile cells, and successfully constructs the human liver microsome and cell co-culture system. Tests prove that the construction method disclosed by the invention can ensure that pollution sources in the human liver microsome cannot influence the sterile environment of the cells, and the sterile state of normal growth of the cells is maintained. Moreover, the method is suitable for in-vitro metabolism research of allogenic substances (such as nicotine and smoke), and lays a foundation for in-vitro metabolism research of the combined action of the human-derived liver microsome and the cells.

The invention provides Ocotillol type esterification derivatives represented by formula (I-R) or formula (I-S) and Ocotillol type esterified derivatives represented by formula (II-R) or formula (II-S). The compounds represented by the formula (II-R) or the formula (II-S) are prepared from the compounds represented by the formula (I-R) or the formula (I-S) through deprotection. The invention further provides a preparation method of the compounds represented by the formula (I-R), the formula (I-S), the formula (II-R) or the formula (II-S) and pharmaceutically acceptable salts of the compounds, and application of the compounds in preparation of anti-inflammatory drugs or anti-inflammatory drug compositions. The Ocotillol type esterified derivatives provided by the invention are obviously superior to the existing clinical drug hydrocortisone sodium succinate in the aspect of inhibiting the generation of an inflammatory signal molecule NO. Moreover, the Ocotillol type esterification derivatives are high in biological friendliness and good in drug safety, show a longer half-life period in in-vitro metabolic stability evaluation, and can improve the safety and stability of the patent drug.

The invention discloses a health food with the effects of removing spots and acne and improving varicosity, which is prepared from (by weight parts) mung bean 2, red bean 2 and black sesame seed 1. The inventive health food has the advantages of simple formulation, easily accessible raw materials, simple preparation method, low cost, and no toxic and adverse effects; and has the effects of promoting in vitro metabolism of free radicals, removing spots and acne and improving varicosity.

The invention provides a kind of reconstructive E.coli CGMCC1667 and reconstructive aminoglycoside enzyme of double functional modification induced by reconstructive E.coli CGMCC1667, and a kind of metabolism model of reconstructive aminoglycoside enzyme of double functional modification, which is disclosed as the application in selecting antibiotic of reconstructive aminoglycoside enzyme of double functional modification and inhibitor of reconstructive aminoglycoside enzyme of double functional modification. This model can research acylation and phosphorylation of antibiotic by reconstructive enzyme of double functional modification. It is charateristic in that it is convenient, quick, accurate and has high degree of automatization, and it can distinct original type and catabolite. It can not only selecting antibiotic of reconstructive aminoglycoside enzyme of double functional modification and inhibitor of reconstructive aminoglycoside enzyme of double functional modification, and also evaluate the stability of new medicine to enzyme of double functional modification.

The invention provides an application of the preclinical drug metabolism and pharmacokinetics key technology and a research system to cefoxitin. The application includes the steps that 1, after cefoxitin in a certain concentration is fed to an experimental animal for a certain time, one or more biological samples are collected from blood, urine and feces; 2, the biological samples obtained in the step 1 are treated with the liquid-liquid extraction technology, the protein precipitation technology and the solid phase extraction technology, and corresponding solutions are prepared; 3, the solutions prepared in the step 2 are analyzed with liquid chromatogram-mass spectrum (LC-MS) and liquid chromatogram-tandem mass spectrum (LC-MS / MS). According to the application, the membrane permeation performance and the cell absorbing capacity of the cefoxitin can be further detected through Caco-2 cells, or detection of the in-vitro metabolism stability and identification of metabolite of the cefoxitin are achieved through the whole animal, S9, human intestine microsome and monoclonal purifying enzymes.

The invention discloses naphthoquinone compound alkannin, which can be used for quantitatively testing the enzyme activity of glucuronic acid transferase UGT1A9 and UGT1A8 as UGT1A9 and UGT1A8 specific probe substrate. The use is implemented by the following step of: selecting alkannin as a specific probe substrate, carrying out the UGT catalytic reaction of a specific substrate by using a UGT in-vitro incubation system, quantitatively testing the enzyme activity of UGT1A9 and UGT1A8 in biological samples and cells by quantitatively testing the lost amount of the substrate or the generated amount of the product. The substrate can be used for quantitative evaluation of the enzyme activity of UGT1A9 / 1A8 in different species, individuals and biological samples, calibration of propofol anesthetic metabolizing capacity of certain tissues, and calibration and estimation of the elimination rate of compounds which are mainly metabolized by UGT1A9 in specific organisms based on in-vitro metabolism dynamic data.

The invention discloses a novel method for studying medicament metabolism by using model organism zebrafish. The method comprises the following steps of: selecting zebrafish as the target for medicament metabolism study, performing experiment grouping design by an optimized experiment grouping method, performing medicament administration in an optimal medicament administration way, extracting medicament metabolites metabolized through the zebrafish by an optimal method, and analyzing the metabolites by an optimal analytical method. The whole experiment method has the advantages of strong operability, capability of objectively reflecting the real metabolism condition of a reaction medicament in vivo, high accuracy of experiment result, capability of overcoming the defect of difficult embodiment of a general in-vitro metabolism experiment in body metabolism comprehensive result and the defects of large dosage and high labour intensity of the general in-vitro metabolism experiment, strong repeatability, small dosage of a tested medicament, low cost, low labour intensity, massive experiment study and high working efficiency.

The invention discloses a shikonin oxime derivative containing an aza-side chain, a preparation method and medical application thereof. The structure of the derivative is shown in a formula (I), the formula (I) is shown in the description, wherein R represents a nitrogen-containing heterocyclic ring; the nitrogen-containing heterocyclic ring comprises a substituent-containing pyridine, pyrimidine,pyrazine, pyridazine, thiazole, benzazole, morpholine, piperidine, piperazine, N-methylene-N-hydroxyethyl piperazine, N-methylene-N-hydroxybutyl piperazine and a physiologically acceptable salt thereof; n is any positive integer ranging from 1 to 4. A compound disclosed by the invention is simple in preparation method, mild in reaction condition, and relatively high in yield. The prepared shikonin naphthazarin parent nucleus hydroxymethylated carbonyl oxime derivative containing the aza-side chain has the advantages of novel structure and good water solubility. Experimental studies on in-vitro metabolism and antitumor activity show that the compound plays a role through a prodrug mechanism and is relatively strong in antitumor activity.

The invention provides application of a preclinical pharmacokinetic key technology and research system in ceftizoxime sodium, comprising the steps of (1) after the ceftizoxime sodium with a certain concentration is administrated to an experimental animal for a certain time, collecting one or more biological samples in blood, urine and faeces; (2) treating the biological samples obtained in step (1) by adopting liquid-liquid extraction, albumen precipitation, and solid phase extraction technologies to obtain a corresponding solution; (3) analyzing the prepared solution in step (2) by adopting an LC-MS(liquid chromatography-mass spectrography) and an LC-MS / MS((liquid chromatography-tandem mass spectrometry). According to the application provided by the invention, the Caco-2 cell can also be adopted to detect the membrane permeability of the medicine and cell absorptive capacity; or the medicine with a certain concentration treats primary culture cerebral microvascular endothelial cells to detect the blood-brain barrier permeability of the medicine; or by adopting a whole animal, S9, human intestinal microsome and monoclonal purified enzyme, in vitro metabolism stability of the medicine is detected and a metabolite is identified.

The invention provides application of a pre-clinical pharmacokinetics key technology and a research system to pemetrexed. The application comprises the steps that 1, after pemetrexed with certain concentration is given to an experimental animal for a certain period of time, one or more biological samples in blood, urine and excrement are collected; 2, the biological samples obtained in the step 1 are treated through liquid-liquid extraction, protein precipitation and solid-phase extraction technologies to be prepared into corresponding solutions; 3, liquid chromatography-mass spectrum (LC-MS) and liquid chromatography-tandem mass spectrum (LC-MS / MS) are adopted for analyzing the solutions prepared in the step 2. According to the application, Caco-2 cells can be adopted for detecting membrane permeating performance and the cell absorbing ability of pemetrexed; or pemetrexed with the certain concentration treats primary culture cerebral microvascular endothelial cells so as to detect blood-brain barrier permeability of pemetrexed; or the overall animal, the overall animal, S9, intestine and liver particle bodies and monoclonal purification enzymes are adopted for detecting the in-vitro metabolism stability of pemetrexed and identifying metabolite.