68 results about "P450 Enzymes" patented technology

Nucleic acids encoding cytochrome P450 variants are provided. The cytochrome P450 variants of have a higher alkane-oxidation capability, alkene-oxidation capability, and/or a higher organic-solvent resistance than the corresponding wild-type or parent cytochrome P450 enzyme. A preferred wild-type cytochrome P450 is cytochrome P450 BM-3. Preferred cytochrome P450 variants include those having an improved capability to hydroxylate alkanes and epoxidate alkenes comprising less than 8 carbons, and have amino acid substitutions corresponding to V78A, H236Q, and E252G of cytochrome P450 BM-3. Preferred cytochrome P450 variants also include those having an improved hydroxylation activity in solutions comprising co-solvents such as DMSO and THF, and have amino acid substitutions corresponding to T235A, R471A, E494K, and S1024E of cytochrome P450 BM-3.

The present invention provides a set of oligonucleotide probe for detecting CYP450 enzyme gene hot mutant site and its uses, belonging to clinical molecular diagnosis field. The probe is designed at whole gene sequence of each subtype enzyme of CYP450 and has relative high sensitivity and specifity. The present invention also provides a gene chip for detecting cytochrome P450 enzyme series mutant sites and can detect gene typing of DNA specimen. The inventive probe can be used in P450 genetype diagnosi, clinical medicament and preventing drug adverse reaction.

Disclosed is a method for detecting CYP450 enzymatic activity by means of micro-liver tissue incubation in vitro, which is characterized in that: a liver trace puncturing method used in clinic takes micro-liver tissues of 0.1 to 0.2g, or kills a mouse to take out the liver in an animal experiment, and builds a micro-liver tissue in vitro incubation system, then uses a one-probe medicine to perform the liquid phase chromatography tandem mass spectrometry for detecting probe substrates and concentration of corresponding metabolites, and obtains the activity of P450 enzyme according to metabolite ratios thereof. The method has the advantages of: 1. micro scale: only micro-liver tissues of 0.1 to 0.2g are needed for detecting cytochrome P450enzyme activity, and can be used for animal experiment and checking clinic micro-liver tissue liver drug enzyme activity; 2. time saving and convenience: costly ultracentrifugation equipment used in the conventional method is saved, costs and time for detection are greatly saved, and liver enzyme metabolic condition simulation is better; and 3. establishing indexes for evaluating the enzyme metabolic activity: metabolite ratios.

Derivatives of dillapiol, sesamol and related monolignans having the following general formula:

These compounds have synergistic properties, inhibit cytochrome P450 enzymes such as human CYP3A4, and can be used as pesticide synergists or pharmaco-enhancers. Accordingly, methods for increasing the efficacy and/or bioavailability of a pharmaceutically active agent and for increasing the potency of a pesticide are described, as are synergistic pesticidal and pharmaceutical compositions.

The invention discloses a primer group and a kit for detecting genetic typing of CYP3A5. The primer group for detecting CYP3A5 gene typing comprises a CY3A5*1 primer pair, a CY3A5*2 primer pair, a CY3A5*3 primer pair, a CY3A5*4 primer pair, a CY3A5*5 primer pair, a CY3A5*6 primer pair and an internal reference primer pair. The application of the kit helps to realize good typing for CYP3A5 gene; and in clinical application of organ transplantation, good guidance effects on individualized use and secure use of immunosuppressant are realized.

This invention relates to a drug-metabolizing enzyme heterogenesis expression system used for out-of-body drug metabolism property evaluation, exactly to say a yeast expression system of coexpressing CYP and CPR, the yeast expression vector which respectively has CPR DNA fragment and CYP DNA fragment is transduced into yeast cell, this two expression vector express in yeast cell by means of occlusal pattern and liberation respectively , custom-crafted yeast cell is fermented and induced, getting yeast cell product which is purpose expression system, it can be used for CYP enzyme system out-of-body drug metabolism property evaluation. The constructed new Saccharomyces cerevisia co-expression system can be extensively used for oxidation reduction enzyme system such as cytopigment P450 enzyme system, the latter one is extensively used for screening, application and development of drug metabolism involved in ADME/T and pharmacokinetics.

The present invention discloses a P450 enzyme of anisodus acutangulus. 4-(1-methyl-2-pyrrolidyl)-3-oxobutanoic acid may be converted into tropinone due to the combined catalysis of the P450 enzyme anda cytochrome P450 reductase. The method disclosed by the present invention is environment-friendly and mild in reaction condition and has an industrial development prospect.

Disclosed herein is a method for treating a patient with Quazepam that reduces the risk of an adverse interaction between the Quazepam and drug that is a substrate of the cytochrome P450 enzyme isoform 2B6 (CYP2B6 substrate drug), e.g., Bupropion. The method includes determining if the patient to be treated with Quazepam is being treated with a CYP2B6 substrate drug, and prescribing or treating the patient with Quazepam based on the determination.

The invention provides an inducing and preparation method of rat liver S9. The inducing and preparation method comprises the following steps of (a) selecting a rat, injecting two types of inducing agents into an abdominal cavity by different types and dosages according to the weight of the rat, and determining the optimum inducing method; (b) killing the rat, removing residual blood in the livers under the sterile condition, and fetching out the livers; (c) under the sterile condition, adding a pre-freezing storage buffer liquid according to a certain liver-wet weight ratio, fully shearing, and equalizing pulp in an ice bath; (d) centrifuging a tissue liquid after pulp equalizing at low speed, collecting a supernatant and a precipitate, centrifuging the supernatant liquid at high speed, and collecting the supernatant liquid, so as to obtain the liver S9 for the first time; (e) adding the pre-freezing storage buffer liquid into the collected precipitate according to a certain ratio, fully grinding under the sterile and ice bath conditions, crushing cells by ultrasonic waves under different conditions, centrifuging at high speed, and collecting the supernatant liquid, so as to obtain the liver S9 again; (f) detecting the contents of protein and enzyme in the liver S9 by a BCA (bicinchoninic acid) method and a CO (carbon monoxide) reduction method under the different conditions. The inducing and preparation method of the rat liver S9 has the advantages that the output of liver S9 is increased, and the content of enzyme P450 is obviously increased.

The present invention relates to a combined pharmaceutical formulation, which is such designed that the release of each ingredient may be controlled to a predetermined release rate by applying the principle of the so-called chronotherapy, where drugs are administered in such a way that the activities of the drugs are expressed at intervals. The formulation of the present invention comprises statin-based lipid-lowering agent and dihydropyridine-based calcium channel blocker that affects cytochrome P450 enzyme as active ingredients, and is such constituted that the release rates of the aforementioned ingredients are different, thus preventing antagonism and side effects, while maintaining the synergistic effect, which leads to the convenience in medication.

The present invention relates to methods for improving the pharmacokinetics of certain compounds useful as inhibitors of the HIV protease enzyme by inhibiting the enzyme activity of cytochrome P450. The present invention also relates to compositions comprising certain compounds useful as inhibitors of the HIV protease enzyme and at least one agent that inhibits the enzyme activity of cytochrome P450.

The invention discloses an NADH analog dependent cytochrome P450 reductase and an application thereof. An NADH analogue is used as a cofactor and reducing power to catalyze electron transfer and reduce cytochrome P450 to complete catalytic circulation. The enzyme is subjected to fusion expression with different types of cytochrome P450 enzymes to construct the NADH analog-dependent, heterozygous and self-sufficient cytochrome P450 enzyme, the obtained NADH analog-dependent cytochrome P450 enzyme can be coupled with oxidoreductases of a regenerated NADH analogue, and the NADH analog is used tocatalyze corresponding substrates of different families of cytochrome P450 to be converted into products. The NADH analog-dependent cytochrome P450 reductase can be used for constructing a biologicalorthogonal metabolic pathway independent of natural cofactor NAD(P)H to realize uncoupling of P450 enzyme-catalyzed energy consumption and endogenous energy metabolism.

The invention provides a cytochrome P450 enzyme mutant and application thereof. Protein modification is carried out on P450 enzyme activity and reverse Markov oxidation selectivity from a Bacillus megaterium wild type strain by means of directed evolution, so that the enzyme activity and selectivity are improved, and a series of P450 enzyme mutants capable of being used for industrial production are developed.

Pharmaceutical compositions are provided that can act as boosters to improve the pharmacokinetics of drugs that undergo in vivo degradation by cytochrome P450 enzymes. Methods of inhibiting cytochrome P450 enzymes are provided that can be used for improving the treatment of diseases by preventing degradation of drugs or other molecules by cytochrome P450. Specifically, methods of inhibiting metabolic degradation of atazanavir sulphate for administering to a patient suffering from HIV infection are disclosed.

The invention discloses a mutation-modified bacillus megaterium ALA2 cytochrome P450 enzyme, and a preparation method and application thereof. The amino acid sequence is shown as SEQ ID NO. 1. Throughdirectional transformation of P450BM-3 monooxygenase genes, the utilization efficiency of recombinant protein to coenzymes NADH is improved greatly by three selected mutation points R966L/K972L/W1046H, 2.4 times of the utilization efficiency of original enzymes to the coenzymes NADH, and the enzyme activity reaches 11415 U/mg.