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Wladyslawa A Daniel - One of the best experts on this subject based on the ideXlab platform.
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Cytochrome P450 2D cyp2D enzyme dysfunction associated with aging and serotonin deficiency in the brain and liver of female dark agouti rats
Neurochemistry International, 2022Co-Authors: Anna Haduch, Przemyslaw J Danek, Wojciech Kuban, Renata Puklo, Natalia Alenina, Joanna Golebiowska, Piotr Popik, Michael Bader, Wladyslawa A DanielAbstract:Among the enzymes that support brain metabolism, Cytochrome P450 (CYP) enzymes occupy an important place. These enzymes catalyze the biotransformation pathways of neuroactive endogenous substrates (neurosteroids, neurotransmitters) and are necessary for the detoxification processes. The aim of the present study was to assess changes in the CYP2D activity and protein level during the aging process and as a result of serotonin deficiency in the female brain. The CYP2D activity was measured in brain and liver microsomes of Dark Agouti wild type (WT) female rats (mature 15-week-old and senescent 18-month-old rats) and in tryptophan hydroxylase 2 (TPH2)-deficient senescent female rats. The CYP2D activity in mature WT Dark Agouti females was independent of the changing phases of the estrous cycle. In senescent WT females rats, the CYP2D activity and protein level were decreased in the cerebral cortex, hippocampus, cerebellum and liver, but increased in the brain stem. In the other examined structures (frontal cortex, hypothalamus, thalamus, striatum), the enzyme activity did not change. In aging TPH2-deficient females, the CYP2D activity and protein levels were decreased in the frontal cortex, hypothalamus and brain stem (activity only), remaining unchanged in other brain structures and liver, relative to senescent WT females. In summary, the aging process and TPH2 deficit affect the CYP2D activity and protein level in female rats, which may have a negative impact on the compensatory capacity of CYP2D in the synthesis of serotonin and dopamine in cerebral structures involved in cognitive and emotional functions. In the liver, the CYP2D-catalyzed drug metabolism may be diminished in elderly females. The results in female rats are compared with those obtained previously in males. It is concluded that aging and serotonin deficiency exert sex-dependent effects on brain CYP2D, which seem to be less favorable in females concerning CYP2D-mediated neurotransmitter synthesis, but beneficial regarding slower neurosteroid metabolism.
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chronic treatment with asenapine affects Cytochrome P450 2D cyp2D in rat brain and liver pharmacological aspects
Neurochemistry International, 2021Co-Authors: Przemyslaw J Danek, Anna Haduch, Ewa Bromek, Wladyslawa A DanielAbstract:Abstract Neuroleptics have to be used for a long time to produce a therapeutic effect. Cytochrome P450 2D (CYP2D) enzymes mediate alternative pathways of neurotransmitter synthesis (i.e. tyramine hydroxylation to dopamine and 5-methoxytryptamine O-demethylation to serotonin), and metabolism of neurosteroids. The aim of our present study was to examine the influence of chronic treatment with the new atypical neuroleptic asenapine on CYP2D in rat brain. In parallel, liver CYP2D was investigated for comparison. Asenapine added in vitro to microsomes of control rats competitively, but weakly inhibited the activity of CYP2D (brain: Ki = 385 μM; liver: Ki = 36 μM). However, prolonged administration of asenapine (0.3 mg/kg sc. for 2 weeks) significantly diminished the activity and protein level of CYP2D in the frontal cortex, nucleus accumbens, hippocampus and cerebellum, but did not affect the enzyme in the hypothalamus, brain stem, substantia nigra and the remainder of the brain. In contrast, asenapine enhanced the enzyme activity and protein level in the striatum. In the liver, chronically administered asenapine reduced the activity and protein level of CYP2D, and the CYP2D1 mRNA level. In conclusion, prolonged administration of asenapine alters the CYP2D expression in the brain structures and in the liver. Through affecting the CYP2D activity in the brain, asenapine may modify its pharmacological effect. By increasing the CYP2D expression/activity in the striatum, asenapine may accelerate the synthesis of dopamine (via tyramine hydroxylation) and serotonin (via 5-methoxytryptamine O-demethylation), and thus alleviate extrapyramidal symptoms. By reducing the CYP2D expression/activity in other brain structures asenapine may diminish the 21-hydroxylation of neurosteroids and thus have a beneficial influence on the symptoms of schizophrenia. In the liver, by reducing the CYP2D activity, asenapine may slow the biotransformation of concomitantly administered CYP2D substrates (drugs) during continuous treatment of schizophrenia or bipolar disorders.
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the activity of brain and liver Cytochrome P450 2D cyp2D is differently affected by antidepressants in the chronic mild stress cms model of depression in the rat
Biochemical Pharmacology, 2018Co-Authors: Anna Haduch, Marta Rysz, Mariusz Papp, Wladyslawa A DanielAbstract:Abstract The effect of two second-generation antidepressants escitalopram and venlafaxine on the activity of brain and liver Cytochrome P450 2D (CYP2D) involved in the metabolism of psychotropics and neurotransmitters was determined in the chronic mild stress (CMS) model of depression. Escitalopram or venlafaxine (10 mg/kg ip/day each) were administered to control and CMS rats for 5 weeks. The activity of CYP2D was studied by measurement of the rate of bufuralol 1′-hydroxylation in microsomes derived from the liver or different brain structures. The obtained results indicate that CMS and the studied antidepressants had different effects on the CYP2D activity depending on the location of the enzyme. In the brain, CMS produced an increase in the CYP2D activity in the hippocampus. Chronic escitalopram or venlafaxine had no effect on the CYP2D activity in the brain of nonstressed rats, however, the antidepressants increased the enzyme activity in the frontal cortex, hypothalamus and cerebellum of stressed animals. In the liver, CMS did not affect the CYP2D activity, while chronic escitalopram or venlafaxine significantly decreased the CYP2D activity and protein level in nonstressed and stressed rats. We conclude that: 1) CMS stimulates the CYP2D activity in the hippocampus and triggers the stimulatory effect of antidepressants on CYP2D in other brain structures; 2) the local brain metabolism of CYP2D substrates (neurosteroids, neurotransmitters, psychotropics) may be enhanced by CMS and/or antidepressants; 3) in contrast to the brain, the liver metabolism of CYP2D substrates may be slower during long-term treatment with escitalopram or venlafaxine.
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the Cytochrome P450 2D mediated formation of serotonin from 5 methoxytryptamine in the brain in vivo a microdialysis study
Journal of Neurochemistry, 2015Co-Authors: Anna Haduch, Ewa Bromek, Marta Kot, Katarzyna Kaminska, Krystyna Golembiowska, Wladyslawa A DanielAbstract:The Cytochrome P450 2D (CYP2D) mediates synthesis of serotonin from 5-methoxytryptamine (5-MT), shown in vitro for cDNA-expressed CYP2D-isoforms and liver and brain microsomes. We aimed to demonstrate this synthesis in the brain in vivo. We measured serotonin tissue content in brain regions after 5-MT injection into the raphe nuclei (Model-A), and its extracellular concentration in rat frontal cortex and striatum using an in vivo microdialysis (Model-B) in male Wistar rats. Naive rats served as control animals. 5-MT injection into the raphe nuclei of PCPA-(tryptophan hydroxylase inhibitor)-pretreated rats increased the tissue concentration of serotonin (from 40 to 90% of the control value, respectively, in the striatum), while the CYP2D inhibitor quinine diminished serotonin level in some brain structures of those animals (Model-A). 5-MT given locally through a microdialysis probe markedly increased extracellular serotonin concentration in the frontal cortex and striatum (to 800 and 1000% of the basal level, respectively) and changed dopamine concentration (Model-B). Quinine alone had no effect on serotonin concentration; however, given jointly with 5-MT, it prevented the 5-MT-induced increase in cortical serotonin in naive rats and in striatal serotonin in PCPA-treated animals. These results indicate that the CYP2D-catalyzed alternative pathway of serotonin synthesis from 5-MT is relevant in the brain in vivo, and set a new target for the action of psychotropics.
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the effect of psychotropic drugs on Cytochrome P450 2D cyp2D in rat brain
European Journal of Pharmacology, 2011Co-Authors: Anna Haduch, Ewa Bromek, Wladyslawa A DanielAbstract:The aim of the study was to investigate the influence of selected antidepressants and neuroleptics on the protein level and activity of Cytochrome P450 2D (CYP2D) in rat brain. The obtained results showed that imipramine, fluoxetine, nefazodone, thioridazine and perazine, added to brain microsomes of control rats, inhibited CYP2D activity to a lower extent (K(i)=255-485μM) than when added to liver microsomes (K(i)=1-45μM), which may result from their stronger affinity for liver CYP2D2 (K(i)=2.7 and 1.25μM for imipramine and fluoxetine, respectively) than for brain CYP2D4 (K(i)=25 and 10μM for imipramine and fluoxetine, respectively), as well as from their high non-specific binding in brain microsomes. Two-week treatment with fluoxetine evoked decreases in the level and activity of CYP2D in the striatum and the nucleus accumbens. In contrast, fluoxetine increased CYP2D expression in the cerebellum, while nefazodone considerably enhanced the activity (but not the protein level) of CYP2D in the truncus cerebri. Imipramine and mirtazapine (active in the liver) did not affect brain CYP2D. Chronic thioridazine decreased CYP2D activity in the substantia nigra and nucleus accumbens, but significantly increased that activity in the striatum and cerebellum. Clozapine significantly enhanced CYP2D activity in the truncus cerebri. In conclusion, psychotropics influence CYP2D in the brain, but their effect is different than in the liver and depends on the cerebral structure. The observed psychotropics-brain CYP2D interactions may be important for the metabolism of neurosteroids and monoaminergic neurotransmitters, and for the local biotransformation of drugs.
Anna Haduch - One of the best experts on this subject based on the ideXlab platform.
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Cytochrome P450 2D cyp2D enzyme dysfunction associated with aging and serotonin deficiency in the brain and liver of female dark agouti rats
Neurochemistry International, 2022Co-Authors: Anna Haduch, Przemyslaw J Danek, Wojciech Kuban, Renata Puklo, Natalia Alenina, Joanna Golebiowska, Piotr Popik, Michael Bader, Wladyslawa A DanielAbstract:Among the enzymes that support brain metabolism, Cytochrome P450 (CYP) enzymes occupy an important place. These enzymes catalyze the biotransformation pathways of neuroactive endogenous substrates (neurosteroids, neurotransmitters) and are necessary for the detoxification processes. The aim of the present study was to assess changes in the CYP2D activity and protein level during the aging process and as a result of serotonin deficiency in the female brain. The CYP2D activity was measured in brain and liver microsomes of Dark Agouti wild type (WT) female rats (mature 15-week-old and senescent 18-month-old rats) and in tryptophan hydroxylase 2 (TPH2)-deficient senescent female rats. The CYP2D activity in mature WT Dark Agouti females was independent of the changing phases of the estrous cycle. In senescent WT females rats, the CYP2D activity and protein level were decreased in the cerebral cortex, hippocampus, cerebellum and liver, but increased in the brain stem. In the other examined structures (frontal cortex, hypothalamus, thalamus, striatum), the enzyme activity did not change. In aging TPH2-deficient females, the CYP2D activity and protein levels were decreased in the frontal cortex, hypothalamus and brain stem (activity only), remaining unchanged in other brain structures and liver, relative to senescent WT females. In summary, the aging process and TPH2 deficit affect the CYP2D activity and protein level in female rats, which may have a negative impact on the compensatory capacity of CYP2D in the synthesis of serotonin and dopamine in cerebral structures involved in cognitive and emotional functions. In the liver, the CYP2D-catalyzed drug metabolism may be diminished in elderly females. The results in female rats are compared with those obtained previously in males. It is concluded that aging and serotonin deficiency exert sex-dependent effects on brain CYP2D, which seem to be less favorable in females concerning CYP2D-mediated neurotransmitter synthesis, but beneficial regarding slower neurosteroid metabolism.
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chronic treatment with asenapine affects Cytochrome P450 2D cyp2D in rat brain and liver pharmacological aspects
Neurochemistry International, 2021Co-Authors: Przemyslaw J Danek, Anna Haduch, Ewa Bromek, Wladyslawa A DanielAbstract:Abstract Neuroleptics have to be used for a long time to produce a therapeutic effect. Cytochrome P450 2D (CYP2D) enzymes mediate alternative pathways of neurotransmitter synthesis (i.e. tyramine hydroxylation to dopamine and 5-methoxytryptamine O-demethylation to serotonin), and metabolism of neurosteroids. The aim of our present study was to examine the influence of chronic treatment with the new atypical neuroleptic asenapine on CYP2D in rat brain. In parallel, liver CYP2D was investigated for comparison. Asenapine added in vitro to microsomes of control rats competitively, but weakly inhibited the activity of CYP2D (brain: Ki = 385 μM; liver: Ki = 36 μM). However, prolonged administration of asenapine (0.3 mg/kg sc. for 2 weeks) significantly diminished the activity and protein level of CYP2D in the frontal cortex, nucleus accumbens, hippocampus and cerebellum, but did not affect the enzyme in the hypothalamus, brain stem, substantia nigra and the remainder of the brain. In contrast, asenapine enhanced the enzyme activity and protein level in the striatum. In the liver, chronically administered asenapine reduced the activity and protein level of CYP2D, and the CYP2D1 mRNA level. In conclusion, prolonged administration of asenapine alters the CYP2D expression in the brain structures and in the liver. Through affecting the CYP2D activity in the brain, asenapine may modify its pharmacological effect. By increasing the CYP2D expression/activity in the striatum, asenapine may accelerate the synthesis of dopamine (via tyramine hydroxylation) and serotonin (via 5-methoxytryptamine O-demethylation), and thus alleviate extrapyramidal symptoms. By reducing the CYP2D expression/activity in other brain structures asenapine may diminish the 21-hydroxylation of neurosteroids and thus have a beneficial influence on the symptoms of schizophrenia. In the liver, by reducing the CYP2D activity, asenapine may slow the biotransformation of concomitantly administered CYP2D substrates (drugs) during continuous treatment of schizophrenia or bipolar disorders.
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the activity of brain and liver Cytochrome P450 2D cyp2D is differently affected by antidepressants in the chronic mild stress cms model of depression in the rat
Biochemical Pharmacology, 2018Co-Authors: Anna Haduch, Marta Rysz, Mariusz Papp, Wladyslawa A DanielAbstract:Abstract The effect of two second-generation antidepressants escitalopram and venlafaxine on the activity of brain and liver Cytochrome P450 2D (CYP2D) involved in the metabolism of psychotropics and neurotransmitters was determined in the chronic mild stress (CMS) model of depression. Escitalopram or venlafaxine (10 mg/kg ip/day each) were administered to control and CMS rats for 5 weeks. The activity of CYP2D was studied by measurement of the rate of bufuralol 1′-hydroxylation in microsomes derived from the liver or different brain structures. The obtained results indicate that CMS and the studied antidepressants had different effects on the CYP2D activity depending on the location of the enzyme. In the brain, CMS produced an increase in the CYP2D activity in the hippocampus. Chronic escitalopram or venlafaxine had no effect on the CYP2D activity in the brain of nonstressed rats, however, the antidepressants increased the enzyme activity in the frontal cortex, hypothalamus and cerebellum of stressed animals. In the liver, CMS did not affect the CYP2D activity, while chronic escitalopram or venlafaxine significantly decreased the CYP2D activity and protein level in nonstressed and stressed rats. We conclude that: 1) CMS stimulates the CYP2D activity in the hippocampus and triggers the stimulatory effect of antidepressants on CYP2D in other brain structures; 2) the local brain metabolism of CYP2D substrates (neurosteroids, neurotransmitters, psychotropics) may be enhanced by CMS and/or antidepressants; 3) in contrast to the brain, the liver metabolism of CYP2D substrates may be slower during long-term treatment with escitalopram or venlafaxine.
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the Cytochrome P450 2D mediated formation of serotonin from 5 methoxytryptamine in the brain in vivo a microdialysis study
Journal of Neurochemistry, 2015Co-Authors: Anna Haduch, Ewa Bromek, Marta Kot, Katarzyna Kaminska, Krystyna Golembiowska, Wladyslawa A DanielAbstract:The Cytochrome P450 2D (CYP2D) mediates synthesis of serotonin from 5-methoxytryptamine (5-MT), shown in vitro for cDNA-expressed CYP2D-isoforms and liver and brain microsomes. We aimed to demonstrate this synthesis in the brain in vivo. We measured serotonin tissue content in brain regions after 5-MT injection into the raphe nuclei (Model-A), and its extracellular concentration in rat frontal cortex and striatum using an in vivo microdialysis (Model-B) in male Wistar rats. Naive rats served as control animals. 5-MT injection into the raphe nuclei of PCPA-(tryptophan hydroxylase inhibitor)-pretreated rats increased the tissue concentration of serotonin (from 40 to 90% of the control value, respectively, in the striatum), while the CYP2D inhibitor quinine diminished serotonin level in some brain structures of those animals (Model-A). 5-MT given locally through a microdialysis probe markedly increased extracellular serotonin concentration in the frontal cortex and striatum (to 800 and 1000% of the basal level, respectively) and changed dopamine concentration (Model-B). Quinine alone had no effect on serotonin concentration; however, given jointly with 5-MT, it prevented the 5-MT-induced increase in cortical serotonin in naive rats and in striatal serotonin in PCPA-treated animals. These results indicate that the CYP2D-catalyzed alternative pathway of serotonin synthesis from 5-MT is relevant in the brain in vivo, and set a new target for the action of psychotropics.
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the effect of psychotropic drugs on Cytochrome P450 2D cyp2D in rat brain
European Journal of Pharmacology, 2011Co-Authors: Anna Haduch, Ewa Bromek, Wladyslawa A DanielAbstract:The aim of the study was to investigate the influence of selected antidepressants and neuroleptics on the protein level and activity of Cytochrome P450 2D (CYP2D) in rat brain. The obtained results showed that imipramine, fluoxetine, nefazodone, thioridazine and perazine, added to brain microsomes of control rats, inhibited CYP2D activity to a lower extent (K(i)=255-485μM) than when added to liver microsomes (K(i)=1-45μM), which may result from their stronger affinity for liver CYP2D2 (K(i)=2.7 and 1.25μM for imipramine and fluoxetine, respectively) than for brain CYP2D4 (K(i)=25 and 10μM for imipramine and fluoxetine, respectively), as well as from their high non-specific binding in brain microsomes. Two-week treatment with fluoxetine evoked decreases in the level and activity of CYP2D in the striatum and the nucleus accumbens. In contrast, fluoxetine increased CYP2D expression in the cerebellum, while nefazodone considerably enhanced the activity (but not the protein level) of CYP2D in the truncus cerebri. Imipramine and mirtazapine (active in the liver) did not affect brain CYP2D. Chronic thioridazine decreased CYP2D activity in the substantia nigra and nucleus accumbens, but significantly increased that activity in the striatum and cerebellum. Clozapine significantly enhanced CYP2D activity in the truncus cerebri. In conclusion, psychotropics influence CYP2D in the brain, but their effect is different than in the liver and depends on the cerebral structure. The observed psychotropics-brain CYP2D interactions may be important for the metabolism of neurosteroids and monoaminergic neurotransmitters, and for the local biotransformation of drugs.
Yoshihiko Funae - One of the best experts on this subject based on the ideXlab platform.
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effect of psychotropic drugs on the 21 hydroxylation of neurosteroids progesterone and allopregnanolone catalyzed by rat cyp2D4 and human cyp2D6 in the brain
Biological & Pharmaceutical Bulletin, 2008Co-Authors: Toshiro Niwa, Toyoko Hiroi, Susumu Imaoka, Shizuo Narimatsu, Kazushi Okada, Yoshihiko FunaeAbstract:We determined the effects of psychotropic drugs on the Cytochrome P450 2D (CYP2D)-mediated 21-hydroxylation of progesterone (PROG) and allopregnanolone (ALLO) with the goal of clarifying whether neurosteroid levels are affected by psychotropic drugs in the brain. PROG or ALLO was incubated with rat CYP2D4 or human CYP2D6 in the presence of typical psychotropic drugs, fluoxetine, imipramine, desipramine, mazindol, and GBR12909, and the 21-hydroxylated metabolites of PROG and ALLO were determined by high performance liquid chromatography and liquid chromatography-tandem mass spectrometry, respectively. Fluoxetine competitively inhibited CYP2D4-mediated PROG 21-hydroxylation and increased both Km and Vmax values of CYP2D6-mediated PROG 21-hydroxylation. In addition, fluoxetine competitively inhibited ALLO 21-hydroxylation mediated by CYP2D4 and CYP2D6. Imipramine, desipramine, mazindol, and GBR12909 competitively inhibited PROG 21-hydroxylation mediated by CYP2D4 and/or CYP2D6, and all psychotropic drugs inhibited ALLO 21-hydroxylation mediated by CYP2D4 and/or CYP2D6. The inhibition constants (Ki values) of imipramine, desipramine, and mazindol against the 21-hydroxylation of PROG and ALLO by CYP2D6 were lower than those by CYP2D4. These results indicate that psychotropic drugs including fluoxetine affected the metabolism of neurosteroids, such as PROG and ALLO in the brain, suggesting that the regulation of the neurosteroid levels is modified by central nervous system-active drugs that inhibit brain CYP2D isoforms.
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Cytochrome P450 2D catalyze steroid 21 hydroxylation in the brain
Endocrinology, 2004Co-Authors: Wataru Kishimoto, Toyoko Hiroi, Susumu Imaoka, Masakazu Shiraishi, Mayuko Osada, Shiro Kominami, Takashi Igarashi, Yoshihiko FunaeAbstract:mRNA of Cytochrome P450 21-hydroxylase (P450c21) is expressed in the brain, but little is known about the enzymatic properties of P450c21 in the brain. In the present study, we showed, by using various recombinant Cytochrome P450 (CYP)2D enzymes and anti-CYP2D4- or P450c21-specific antibodies, that rat brain microsomal steroid 21-hydroxylation is catalyzed not by P450c21, but by CYP2D isoforms. Rat CYP2D4 and human CYP2D6, which are the predominant CYP2D isoforms in the brain, possess 21-hydroxylation activity for both progesterone and 17alpha-hydroxyprogesterone. In rat brain microsomes, these activities were not inhibited by anti-P450c21 antibodies, but they were effectively inhibited by the CYP2D-specific chemical inhibitor quinidine and by anti-CYP2D4 antibodies. mRNA and protein of CYP2D4 were expressed throughout the brain, especially in cerebellum, striatum, pons, and medulla oblongata, whereas the mRNA and protein levels of P450c21 were extremely low or undetectable. These results support the idea that CYP2D4, not P450c21, works as steroid 21-hydroxylase in the brain. Allopregnanolone, a representative gamma-aminobutyric acid receptor modulator, was also hydroxylated at the C-21 position by recombinant CYP2D4 and CYP2D6. Rat brain microsomal allopregnanolone 21-hydroxylation was inhibited by fluoxetine with an IC(50) value of 2 microm, suggesting the possibility that the brain CYP2D isoforms regulate levels of neurosteroids such as allopregnanolone, and that this regulation is modified by central nervous system-active drugs such as fluoxetine.
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homology modeling of rat and human Cytochrome P450 2D cyp2D isoforms and computational rationalization of experimental ligand binding specificities
Journal of Medicinal Chemistry, 2003Co-Authors: Jennifer Venhorst, Toyoko Hiroi, Yoshihiko Funae, Antonius Ter Laak, Jan N M Commandeur, Nico P E VermeulenAbstract:The ligand-binding characteristics of rat and human CYP2D isoforms, i.e., rat CYP2D1−4 and human CYP2D6, were investigated by measuring IC50 values of 11 known CYP2D6 ligands using 7-methoxy-4-(aminomethyl)coumarin (MAMC) as substrate. Like CYP2D6, all rat CYP2D isozymes catalyzed the O-demethylation of MAMC with Km and Vmax values ranging between 78 and 145 μM and 0.048 and 1.122 min-1, respectively. To rationalize observed differences in the experimentally determined IC50 values, homology models of the CYP2D isoforms were constructed. A homology model of CYP2D6 was generated on the basis of crystallized rabbit CYP2C5 and was validated on its ability to reproduce binding orientations corresponding to metabolic profiles of the substrates and to remain stable during unrestrained molecular dynamics simulations at 300 K. Twenty-two active site residues, sharing up to 59% sequence identity, were identified in the CYP2D binding pockets and included CYP2D6 residues Phe120, Glu216, and Asp301. Electrostatic pote...
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progesterone oxidation by Cytochrome P450 2D isoforms in the brain
Endocrinology, 2001Co-Authors: Toyoko Hiroi, Susumu Imaoka, Wataru Kishimoto, Takashi Igarashi, Toshio Chow, Yoshihiko FunaeAbstract:The existence of Cytochrome P450 2D isoforms in the brain has been demonstrated, although their physiological functions remain to be elucidated. In this study we demonstrated that recombinant rat Cytochrome P450 2D1 and 2D4 and human Cytochrome P450 2D6 possess progesterone 6β- and 16α- hydroxylation activities; 2β- and 21-hydroxylation activities; and 2β-, 6β-, 16α- and 21-hydroxylation activities, respectively. Cytochrome P450 2D4 had the lowest Km value and the highest maximum velocity value toward these activities. Progesterone 2β- and 21-hydroxylation activities were also detected in rat brain microsomes, and these activities were completely inhibited by antiCytochrome P450 2D antibodies. The presence of endogenous 2β- and 21-hydroxyprogesterones in rat brain tissues was also demonstrated. The mRNAs of Cytochrome P450 2D4, CYP11A, and 3β-hydroxysteroid dehydrogenase were detected in the rat brain, suggesting that progesterone was generated from cholesterol by CYP11A and 3β-hydroxysteroid dehydrogenas...
Toyoko Hiroi - One of the best experts on this subject based on the ideXlab platform.
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effect of psychotropic drugs on the 21 hydroxylation of neurosteroids progesterone and allopregnanolone catalyzed by rat cyp2D4 and human cyp2D6 in the brain
Biological & Pharmaceutical Bulletin, 2008Co-Authors: Toshiro Niwa, Toyoko Hiroi, Susumu Imaoka, Shizuo Narimatsu, Kazushi Okada, Yoshihiko FunaeAbstract:We determined the effects of psychotropic drugs on the Cytochrome P450 2D (CYP2D)-mediated 21-hydroxylation of progesterone (PROG) and allopregnanolone (ALLO) with the goal of clarifying whether neurosteroid levels are affected by psychotropic drugs in the brain. PROG or ALLO was incubated with rat CYP2D4 or human CYP2D6 in the presence of typical psychotropic drugs, fluoxetine, imipramine, desipramine, mazindol, and GBR12909, and the 21-hydroxylated metabolites of PROG and ALLO were determined by high performance liquid chromatography and liquid chromatography-tandem mass spectrometry, respectively. Fluoxetine competitively inhibited CYP2D4-mediated PROG 21-hydroxylation and increased both Km and Vmax values of CYP2D6-mediated PROG 21-hydroxylation. In addition, fluoxetine competitively inhibited ALLO 21-hydroxylation mediated by CYP2D4 and CYP2D6. Imipramine, desipramine, mazindol, and GBR12909 competitively inhibited PROG 21-hydroxylation mediated by CYP2D4 and/or CYP2D6, and all psychotropic drugs inhibited ALLO 21-hydroxylation mediated by CYP2D4 and/or CYP2D6. The inhibition constants (Ki values) of imipramine, desipramine, and mazindol against the 21-hydroxylation of PROG and ALLO by CYP2D6 were lower than those by CYP2D4. These results indicate that psychotropic drugs including fluoxetine affected the metabolism of neurosteroids, such as PROG and ALLO in the brain, suggesting that the regulation of the neurosteroid levels is modified by central nervous system-active drugs that inhibit brain CYP2D isoforms.
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Cytochrome P450 2D catalyze steroid 21 hydroxylation in the brain
Endocrinology, 2004Co-Authors: Wataru Kishimoto, Toyoko Hiroi, Susumu Imaoka, Masakazu Shiraishi, Mayuko Osada, Shiro Kominami, Takashi Igarashi, Yoshihiko FunaeAbstract:mRNA of Cytochrome P450 21-hydroxylase (P450c21) is expressed in the brain, but little is known about the enzymatic properties of P450c21 in the brain. In the present study, we showed, by using various recombinant Cytochrome P450 (CYP)2D enzymes and anti-CYP2D4- or P450c21-specific antibodies, that rat brain microsomal steroid 21-hydroxylation is catalyzed not by P450c21, but by CYP2D isoforms. Rat CYP2D4 and human CYP2D6, which are the predominant CYP2D isoforms in the brain, possess 21-hydroxylation activity for both progesterone and 17alpha-hydroxyprogesterone. In rat brain microsomes, these activities were not inhibited by anti-P450c21 antibodies, but they were effectively inhibited by the CYP2D-specific chemical inhibitor quinidine and by anti-CYP2D4 antibodies. mRNA and protein of CYP2D4 were expressed throughout the brain, especially in cerebellum, striatum, pons, and medulla oblongata, whereas the mRNA and protein levels of P450c21 were extremely low or undetectable. These results support the idea that CYP2D4, not P450c21, works as steroid 21-hydroxylase in the brain. Allopregnanolone, a representative gamma-aminobutyric acid receptor modulator, was also hydroxylated at the C-21 position by recombinant CYP2D4 and CYP2D6. Rat brain microsomal allopregnanolone 21-hydroxylation was inhibited by fluoxetine with an IC(50) value of 2 microm, suggesting the possibility that the brain CYP2D isoforms regulate levels of neurosteroids such as allopregnanolone, and that this regulation is modified by central nervous system-active drugs such as fluoxetine.
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homology modeling of rat and human Cytochrome P450 2D cyp2D isoforms and computational rationalization of experimental ligand binding specificities
Journal of Medicinal Chemistry, 2003Co-Authors: Jennifer Venhorst, Toyoko Hiroi, Yoshihiko Funae, Antonius Ter Laak, Jan N M Commandeur, Nico P E VermeulenAbstract:The ligand-binding characteristics of rat and human CYP2D isoforms, i.e., rat CYP2D1−4 and human CYP2D6, were investigated by measuring IC50 values of 11 known CYP2D6 ligands using 7-methoxy-4-(aminomethyl)coumarin (MAMC) as substrate. Like CYP2D6, all rat CYP2D isozymes catalyzed the O-demethylation of MAMC with Km and Vmax values ranging between 78 and 145 μM and 0.048 and 1.122 min-1, respectively. To rationalize observed differences in the experimentally determined IC50 values, homology models of the CYP2D isoforms were constructed. A homology model of CYP2D6 was generated on the basis of crystallized rabbit CYP2C5 and was validated on its ability to reproduce binding orientations corresponding to metabolic profiles of the substrates and to remain stable during unrestrained molecular dynamics simulations at 300 K. Twenty-two active site residues, sharing up to 59% sequence identity, were identified in the CYP2D binding pockets and included CYP2D6 residues Phe120, Glu216, and Asp301. Electrostatic pote...
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progesterone oxidation by Cytochrome P450 2D isoforms in the brain
Endocrinology, 2001Co-Authors: Toyoko Hiroi, Susumu Imaoka, Wataru Kishimoto, Takashi Igarashi, Toshio Chow, Yoshihiko FunaeAbstract:The existence of Cytochrome P450 2D isoforms in the brain has been demonstrated, although their physiological functions remain to be elucidated. In this study we demonstrated that recombinant rat Cytochrome P450 2D1 and 2D4 and human Cytochrome P450 2D6 possess progesterone 6β- and 16α- hydroxylation activities; 2β- and 21-hydroxylation activities; and 2β-, 6β-, 16α- and 21-hydroxylation activities, respectively. Cytochrome P450 2D4 had the lowest Km value and the highest maximum velocity value toward these activities. Progesterone 2β- and 21-hydroxylation activities were also detected in rat brain microsomes, and these activities were completely inhibited by antiCytochrome P450 2D antibodies. The presence of endogenous 2β- and 21-hydroxyprogesterones in rat brain tissues was also demonstrated. The mRNAs of Cytochrome P450 2D4, CYP11A, and 3β-hydroxysteroid dehydrogenase were detected in the rat brain, suggesting that progesterone was generated from cholesterol by CYP11A and 3β-hydroxysteroid dehydrogenas...
Ewa Bromek - One of the best experts on this subject based on the ideXlab platform.
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chronic treatment with asenapine affects Cytochrome P450 2D cyp2D in rat brain and liver pharmacological aspects
Neurochemistry International, 2021Co-Authors: Przemyslaw J Danek, Anna Haduch, Ewa Bromek, Wladyslawa A DanielAbstract:Abstract Neuroleptics have to be used for a long time to produce a therapeutic effect. Cytochrome P450 2D (CYP2D) enzymes mediate alternative pathways of neurotransmitter synthesis (i.e. tyramine hydroxylation to dopamine and 5-methoxytryptamine O-demethylation to serotonin), and metabolism of neurosteroids. The aim of our present study was to examine the influence of chronic treatment with the new atypical neuroleptic asenapine on CYP2D in rat brain. In parallel, liver CYP2D was investigated for comparison. Asenapine added in vitro to microsomes of control rats competitively, but weakly inhibited the activity of CYP2D (brain: Ki = 385 μM; liver: Ki = 36 μM). However, prolonged administration of asenapine (0.3 mg/kg sc. for 2 weeks) significantly diminished the activity and protein level of CYP2D in the frontal cortex, nucleus accumbens, hippocampus and cerebellum, but did not affect the enzyme in the hypothalamus, brain stem, substantia nigra and the remainder of the brain. In contrast, asenapine enhanced the enzyme activity and protein level in the striatum. In the liver, chronically administered asenapine reduced the activity and protein level of CYP2D, and the CYP2D1 mRNA level. In conclusion, prolonged administration of asenapine alters the CYP2D expression in the brain structures and in the liver. Through affecting the CYP2D activity in the brain, asenapine may modify its pharmacological effect. By increasing the CYP2D expression/activity in the striatum, asenapine may accelerate the synthesis of dopamine (via tyramine hydroxylation) and serotonin (via 5-methoxytryptamine O-demethylation), and thus alleviate extrapyramidal symptoms. By reducing the CYP2D expression/activity in other brain structures asenapine may diminish the 21-hydroxylation of neurosteroids and thus have a beneficial influence on the symptoms of schizophrenia. In the liver, by reducing the CYP2D activity, asenapine may slow the biotransformation of concomitantly administered CYP2D substrates (drugs) during continuous treatment of schizophrenia or bipolar disorders.
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the Cytochrome P450 2D mediated formation of serotonin from 5 methoxytryptamine in the brain in vivo a microdialysis study
Journal of Neurochemistry, 2015Co-Authors: Anna Haduch, Ewa Bromek, Marta Kot, Katarzyna Kaminska, Krystyna Golembiowska, Wladyslawa A DanielAbstract:The Cytochrome P450 2D (CYP2D) mediates synthesis of serotonin from 5-methoxytryptamine (5-MT), shown in vitro for cDNA-expressed CYP2D-isoforms and liver and brain microsomes. We aimed to demonstrate this synthesis in the brain in vivo. We measured serotonin tissue content in brain regions after 5-MT injection into the raphe nuclei (Model-A), and its extracellular concentration in rat frontal cortex and striatum using an in vivo microdialysis (Model-B) in male Wistar rats. Naive rats served as control animals. 5-MT injection into the raphe nuclei of PCPA-(tryptophan hydroxylase inhibitor)-pretreated rats increased the tissue concentration of serotonin (from 40 to 90% of the control value, respectively, in the striatum), while the CYP2D inhibitor quinine diminished serotonin level in some brain structures of those animals (Model-A). 5-MT given locally through a microdialysis probe markedly increased extracellular serotonin concentration in the frontal cortex and striatum (to 800 and 1000% of the basal level, respectively) and changed dopamine concentration (Model-B). Quinine alone had no effect on serotonin concentration; however, given jointly with 5-MT, it prevented the 5-MT-induced increase in cortical serotonin in naive rats and in striatal serotonin in PCPA-treated animals. These results indicate that the CYP2D-catalyzed alternative pathway of serotonin synthesis from 5-MT is relevant in the brain in vivo, and set a new target for the action of psychotropics.
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the effect of psychotropic drugs on Cytochrome P450 2D cyp2D in rat brain
European Journal of Pharmacology, 2011Co-Authors: Anna Haduch, Ewa Bromek, Wladyslawa A DanielAbstract:The aim of the study was to investigate the influence of selected antidepressants and neuroleptics on the protein level and activity of Cytochrome P450 2D (CYP2D) in rat brain. The obtained results showed that imipramine, fluoxetine, nefazodone, thioridazine and perazine, added to brain microsomes of control rats, inhibited CYP2D activity to a lower extent (K(i)=255-485μM) than when added to liver microsomes (K(i)=1-45μM), which may result from their stronger affinity for liver CYP2D2 (K(i)=2.7 and 1.25μM for imipramine and fluoxetine, respectively) than for brain CYP2D4 (K(i)=25 and 10μM for imipramine and fluoxetine, respectively), as well as from their high non-specific binding in brain microsomes. Two-week treatment with fluoxetine evoked decreases in the level and activity of CYP2D in the striatum and the nucleus accumbens. In contrast, fluoxetine increased CYP2D expression in the cerebellum, while nefazodone considerably enhanced the activity (but not the protein level) of CYP2D in the truncus cerebri. Imipramine and mirtazapine (active in the liver) did not affect brain CYP2D. Chronic thioridazine decreased CYP2D activity in the substantia nigra and nucleus accumbens, but significantly increased that activity in the striatum and cerebellum. Clozapine significantly enhanced CYP2D activity in the truncus cerebri. In conclusion, psychotropics influence CYP2D in the brain, but their effect is different than in the liver and depends on the cerebral structure. The observed psychotropics-brain CYP2D interactions may be important for the metabolism of neurosteroids and monoaminergic neurotransmitters, and for the local biotransformation of drugs.
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the ability of Cytochrome P450 2D isoforms to synthesize dopamine in the brain an in vitro study
European Journal of Pharmacology, 2010Co-Authors: Ewa Bromek, Anna Haduch, Wladyslawa A DanielAbstract:The present study was aimed at determining which rat Cytochrome P450 (CYP) isoforms are involved in the hydroxylation of tyramine to dopamine and at determining whether the reaction can take place in the brain. Moreover, we examined the relative distribution of the CYP2D subfamily's activity in the rat brain, focusing our attention on dopaminergic structures. The study was conducted with cDNA-expressed CYP isoforms (rat CYP1A1, 2A2, 2B1, 2C6/11/13, 2D1/2/4/18, 2E1, 3A2 and human CYP2D6) and with rat brain microsomes. Of the rat CYP isoforms tested, only CYP2D2, 2D4 and 2D18 (but not CYP2D1) were capable of forming dopamine from tyramine. The rat CYP2D isoforms were less efficient than the human CYP2D6 and the efficiency of both human and rat enzymes was higher for m-tyramine (K(m)=256, 143 and 87 microM; V(max)=0.47, 0.23 and 9.55 pmol/pmolCYP/min for CYP2D4, 2D18 and 2D6, respectively) than for p-tyramine (K(m)=433 and 688 microM, V(max)=0.12 and 0.19 pmol/pmolCYP/min for CYP2D4 and 2D18, respectively). Brain microsomes were able to metabolize tyramine to dopamine. The reaction was inhibited by the CYP2D inhibitor quinine and by anti-CYP2D4 antibodies, which suggests that CYP2D4 is the isoform governing tyramine hydroxylation to dopamine in the rat brain. A relatively high level of CYP2D activity (bufuralol 1'-hydroxylation) was found in the substantia nigra, the cerebellum, the nucleus accumbens and the truncus cerebri. The results are discussed in the context of the likelihood of CYP2D-mediated dopamine synthesis in vivo, the implications for Parkinson's disease and the addiction process.
