68 results about "P450 Enzymes" patented technology

The present invention provides methods and systems or apparatuses, to analyze multiple molecular and clinical variables from an individual diagnosed with a psychiatric disorder, such as post-traumatic stress disorder (PTSD), in order to optimize medication selection for therapeutic response. Molecular co-variables include polymorphisms in genes including those involved in central control and mediation of the hypothalamic-pituitary axis (HPA) stress response, the density of methylation in regulatory regions of said polymorphic genes, polymorphisms in genes that encode cytochrome P450 enzymes responsible for drug metabolism, and drug-drug and drug-gene interactions. Clinical co-variables include but are not limited to the sex, age and ethnicity of that individual, medication history, family history, diagnostic codes, Pittsburgh insomnia rating score, and Charlson index score. The system makes a determination based on unstructured and structured data types derived from internal and external knowledge resources to determine psychotropic drug choice that best matches the molecular and clinical variation profile of an individual patient. The decision support system provides a therapeutic recommendation for a clinician based on the patient's variation profile.

The present application provides methods and compositions for improving the bioavailability of a lipid-containing antiviral compound, and in particular, an antiviral lipid-containing compound. In one embodiment, pharmaceutically acceptable compositions are provided that include an antiviral lipid-containing compound, or salt, ester, or prodrug thereof and one or more bioavailability enhancing compounds, such as inhibitors of cytochrome P450 enzymes.

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.

The present invention provides methods, compositions, substrates, and kits useful for analyzing the metabolic activity in cells, tissue, and animals and for screening test compounds for their effect on cytochrome P450 activity. In particular, a one-step and two-step methods using luminogenic molecules, e.g. luciferin or coelenterazines, that are cytochrome P450 substrates and that are also bioluminescent enzyme, e.g., luciferase, pro-substrates are provided. Upon addition of the luciferin derivative or other luminogenic molecule into a P450 reaction, the P450 enzyme metabolizes the molecule into a bioluminescent enzyme substrate, e.g., luciferin and / or luciferin derivative metabolite, in a P450 reaction. The resulting metabolite(s) serves as a substrate of the bioluminescent enzyme, e.g., luciferase, in a second light-generating reaction. Luminescent cytochrome P450 assays with low background signals and high sensitivity are disclosed and isoform selectivity is demonstrated. The present invention also provides an improved method for performing luciferase reactions which employs added pyrophosphatase to remove inorganic pyrophosphate, a luciferase inhibitor which may be present in the reaction mixture as a contaminant or may be generated during the reaction. The present method further provides a method for stabilizing and prolonging the luminescent signal in a luciferase-based assay using luciferase stabilizing agents such as reversible luciferase inhibitors.

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.

Methods are disclosed for developing models used to rapidly predict metabolic stability and regioselectivity of drug molecules. Training sets, based on a sample of molecules with known reaction rates and / or activation energies, are used along with structural descriptors of the molecules in order to develop mathematical models of metabolism based on regression analysis of the activation energies and descriptors. The resulting models are then used to predict the metabolism of other molecules. The invention is particularly useful in developing simple models of cytochrome p450 enzyme metabolism.

The present invention relates to p450 enzymes and nucleic acid sequences encoding p450 enzymes in Nicotiana, and methods of using those enzymes and nucleic acid sequences to alter plant phenotypes.

A method of regulating the activity of human cytochrome P450 isozyme CYP2A6 to control nicotine metabolism or decrease to production of carcinogens from procarcinogens, such as those present in tobacco smoke, in an individual by selectively inhibiting CYP2A6. Various prophylactic (i.e., prevention and treatment) compositions and methods are also described, including an improved oral nicotine composition and method comprising the use of nicotine together with an inhibitor of the CYP2A6 enzyme. Furthermore, it has been discovered that the presence in an individual of a mutant allele of human cytochrome P450 enzyme CYP2A6 (referred to throughout this specification as “CYP2A6” for brevity) is predictive of an individual who: (i) has a decreased risk of becoming a smoker, (ii) will smoke less if he / she becomes dependent, and / or (iii) may be at relatively lower risk for cancer due to both decreased smoke exposure and decreased CYP2A6-mediated activation of tobacco smoke and other procarcinogenic substrates. This invention provides diagnostic methods for predicting tobacco dependence risk and risk for cancers related to CYP2A6 substrates in an individual by analysing for the presence of a mutant genotype for human cytochrome P450 enzyme CYP2A6 in an individual, ranging from gene duplication (multiple copies of CYP2A6) to single or even no copies due to null alleles or gene deletion.

The present invention relates to the production of chemical species in bacterial host cells. Particularly, the present invention provides for the production of chemical species in Escherichia coli (E. coli) host cells that functionally express engineered P450 enzymes.

The product relates to Bacillus megaterium ALA2 cytochrome P450 enzyme gene and methods of recombinant plasmid construction and enzyme purification and the nucleotide sequence cypI of the gene is mentioned in SEQIDNO.1. Through escherichia coli expression carrier selection and gene directed modification, the expression level of recombinant protein substantially is increased by a selected pTrc99A carrier and is 3.8 times higher than the expression level of recombinant protein with a selected pET20b carrier, and the enzyme activity reaches 700U / mL. The modified coding Bacillus megaterium ALA2 cytochromes P450 enzyme gene cypI is inserted in pYrc99A and transformed into escherichia coli. Then the enzyme activity expressed in escherichia coli raises 32% in comparison with that of original gene cyp and reaches 1020U / mL (figure-2).

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.

A method of regulating the activity of human cytochrome P450 isozyme CYP2A6 to control nicotine metabolism or decrease the production of carcinogens from procarcinogens, such as those present in tobacco smoke, in an individual by selectively inhibiting CYP2A6. Various prophylactic (i.e., prevention and treatment) compositions and methods are also described, including an improved oral nicotine composition and method comprising the use of nicotine together with an inhibitor of the CYP2A6 enzyme. Furthermore, it has been discovered that the presence in an individual of a mutant allele of human cytochrome P450 enzyme CYP2A6 (referred to throughout this specification as 'CYP2A6' for brevity) is predictive of an individual who: (1) has a decreased risk of becoming a smoker, (ii) will smoke less if he / she becomes dependent, and / or (iii) may be at relatively lower risk for cancer due to both decreased smoke exposure and decreased CYP2A6 -mediated activation of tobacco smoke and other procarcinogenic substrates. This invention provides diagnostic methods for predicting tobacco dependence risk and risk for cancers related to CYP2A6 substrates in an individual by analyzing for the presence of a mutant genotype for human cytochrome P450 enzyme CYP2A6 in an individual, ranging from gene duplication (multiple copies of CYP2A6) to single or even no copies due to null alleles or gene deletion.

A method of regulating the activity of human cytochrome P450 isozyme CYP2A6 to control nicotine metabolism or decrease the production of carcinogens from procarcinogens, such as those present in tobacco smoke, in an individual by selectively inhibiting CYP2A6. Various prophylactic (i.e., prevention and treatment) compositions and methods are also described, including an improved oral nicotine composition and method comprising the use of nicotine together with an inhibitor of the CYP2A6 enzyme. Furthermore, it has been discovered that the presence in an individual of a mutant allele of human cytochrome P450 enzyme CYP2A6 (referred to throughout this specification as “CYP2A6” for brevity) is predictive of an individual who: (1) has a decreased risk of becoming a smoker, (ii) will smoke less if he / she becomes dependent, and / or (iii) may be at relatively lower risk for cancer due to both decreased smoke exposure and decreased CYP2A6 -mediated activation of tobacco smoke and other procarcinogenic substrates. This invention provides diagnostic methods for predicting tobacco dependence risk and risk for cancers related to CYP2A6 substrates in an individual by analyzing for the presence of a mutant genotype for human cytochrome P450 enzyme CYP2A6 in an individual, ranging from gene duplication (multiple copies of CYP2A6) to single or even no copies due to null alleles or gene deletion.

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 invention related to 3-alkoxy-substituent-2-pyrazinyl formamide compounds with a structure represented by the general formula I, and an application thereof in preparing antiviral medicines. The compounds provided by the invention perform antiviral effects in vivo through a process of being metabolized into T1105 or T705 by esterase or P450 enzyme. Compared with prototype medicines T1105 or T705, the compounds with the general formula I have substantial advantages of high bioavailability and long in-vivo action time.

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.

The present disclosure teaches that the recombination of homologous sequences of P450 enzymes, with the aid of SCHEMA to predict a resulting protein structure, is able to generate libraries of chimeras with significant functional diversity. Additionally, the members of these libraries demonstrate superior or unexpected new properties, which correlate with other factors that are observable in the library. Thus, the making of libraries of optimized P450 enzymes, the analysis of libraries to identify an optimized subset, and the optimized chimeras with improved or altered functionalities are all taught in the present disclosure.

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.

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 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.

A method of regulating the activity of human cytochrome P450 isozyme CYP2A6 to control nicotine metabolism or decrease to production of carcinogens from procarcinogens, such as those present in tobacco smoke, in an individual by selectively inhibiting CYP2A6. Various prophylactic (i.e., prevention and treatment) compositions and methods are also described, including an improved oral nicotine composition and method comprising the use of nicotine together with an inhibitor of the CYP2A6 enzyme.Furthermore, it has been discovered that the presence in an individual of a mutant allele of human cytochrome P450 enzyme CYP2A6 (referred to throughout this specification as “CYP2A6” for brevity) is predictive of an individual who: (i) has a decreased risk of becoming a smoker, (ii) will smoke less if he / she becomes dependent, and / or (iii) may be at relatively lower risk for cancer due to both decreased smoke exposure and decreased CYP2A6-mediated activation of tobacco smoke and other procarcinogenic substrates. This invention provides diagnostic methods for predicting tobacco dependence risk and risk for cancers related to CYP2A6 substrates in an individual by analysing for the presence of a mutant genotype for human cytochrome P450 enzyme CYP2A6 in an individual, ranging from gene duplication (multiple copies of CYP2A6) to single or even no copies due to null alleles or gene deletion.