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1.
Biomed Chromatogr ; 36(10): e5439, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35778888

RESUMO

To evaluate the effect of imrecoxib on CYP2C11 enzyme activity, mRNA, and protein expression, a UPLC method was established. Tolbutamide was selected as the CYP2C11 enzyme-specific probe drug and incubated with imrecoxib in rat liver microsomes. The yield of 4-hydroxytolbutamide was measured using UPLC to investigate the effect of imrecoxib on CYP2C11 enzyme activity. Imrecoxib (10 mg/kg) was administered intragastrically twice daily. After 1, 7, and 14 days of administration, the liver tissues were analyzed. The expression of CYP2C11 enzyme mRNA was determined using reverse transcription-polymerase chain reaction, and its protein expression was determined using Western blot analysis. Imrecoxib concentration was inversely proportional to the production of 4-hydroxytolbutamide in liver microsomes. Imrecoxib demonstrated a dose-dependent inhibitory effect on CYP2C11 activity with IC50 = 74.77 µM. After administration, reverse transcription-polymerase chain reaction showed CYP2C11 enzyme mRNA expressions were 65% (P < 0.05), 35%, and 34% of the control group, respectively (P < 0.01). Western blot analysis showed CYP2C11 enzyme protein expressions were 80, 37, and 34% of the control group, respectively (P < 0.01). Imrecoxib can reduce mRNA and protein expression of CYP2C11 enzyme in rat liver and inhibit the activity of CYP2C11 enzyme in a dose-dependent manner. However, it does not produce clinically significant drug interactions.


Assuntos
Hidrocarboneto de Aril Hidroxilases , Pirróis , Animais , Hidrocarboneto de Aril Hidroxilases/metabolismo , Família 2 do Citocromo P450/genética , Família 2 do Citocromo P450/metabolismo , Microssomos Hepáticos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Mensageiro/farmacologia , Ratos , Esteroide 16-alfa-Hidroxilase/genética , Esteroide 16-alfa-Hidroxilase/metabolismo , Sulfetos
2.
Biochem Pharmacol ; 195: 114850, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34822809

RESUMO

Arachidonic acid (AA)-derived cytochrome P450 (CYP) derivatives, epoxyeicosatrienoic acids (EETs) and 20-hidroxyeicosatetranoic acid (20-HETE), play a key role in kidney tubular and vascular functions and blood pressure. Altered metabolism of CYP epoxygenases and CYP hydroxylases has differentially been involved in the pathogenesis of metabolic disease-associated vascular complications, although the mechanisms responsible for the vascular injury are unclear. The present study aimed to assess whether obesity-induced changes in CYP enzymes may contribute to oxidative stress and endothelial dysfunction in kidney preglomerular arteries. Endothelial function and reactive oxygen species (ROS) production were assessed in interlobar arteries of obese Zucker rats (OZR) and their lean counterparts lean Zucker rats (LZR) and the effects of CYP2C and CYP4A inhibitors sulfaphenazole and HET0016, respectively, were examined on the endothelium-dependent relaxations and O2- and H2O2 levels of preglomerular arteries. Non-nitric oxide (NO) non-prostanoid endothelium-derived hyperpolarization (EDH)-type responses were preserved but resistant to the CYP epoxygenase blocker sulfaphenazole in OZR in contrast to those in LZR. Sulfaphenazole did not further inhibit reduced arterial H2O2 levels, and CYP2C11/CYP2C23 enzymes were downregulated in intrarenal arteries from OZR. Renal EDH-mediated relaxations were preserved in obese rats by the enhanced activity and expression of endothelial calcium-activated potassium channels (KCa). CYP4A blockade restored impaired NO-mediated dilatation and inhibited augmented O2- production in kidney arteries from OZR. The current data demonstrate that both decreased endothelial CYP2C11/ CYP2C23-derived vasodilator H2O2 and augmented CYP4A-derived 20-HETE contribute to endothelial dysfunction and vascular oxidative stress in obesity. CYP4A inhibitors ameliorate arterial oxidative stress and restore endothelial function which suggests its therapeutic potential for the vascular complications of obesity-associated kidney injury.


Assuntos
Sistema Enzimático do Citocromo P-450/metabolismo , Endotélio Vascular/metabolismo , Rim/metabolismo , Obesidade/metabolismo , Estresse Oxidativo , Artéria Renal/metabolismo , Amidinas/farmacologia , Animais , Hidrocarboneto de Aril Hidroxilases/metabolismo , Citocromo P-450 CYP4A/metabolismo , Família 2 do Citocromo P450/metabolismo , Peróxido de Hidrogênio/metabolismo , Ácidos Hidroxieicosatetraenoicos/antagonistas & inibidores , Ácidos Hidroxieicosatetraenoicos/metabolismo , Rim/irrigação sanguínea , Masculino , Obesidade/fisiopatologia , Ratos Zucker , Espécies Reativas de Oxigênio/metabolismo , Artéria Renal/efeitos dos fármacos , Artéria Renal/fisiopatologia , Esteroide 16-alfa-Hidroxilase/metabolismo , Sulfafenazol/farmacologia , Vasodilatação/efeitos dos fármacos
3.
J Vet Med Sci ; 83(8): 1338-1344, 2021 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-34176823

RESUMO

Borneol is a traditional Chinese medicine. In Chinese veterinary clinics, borneol and its related compounds are often used in combination with florfenicol to treat respiratory infections. This study investigated whether the pharmacokinetics of florfenicol in rats was affected by its concomitant use with borneol. Sprague-Dawley rats were intragastrically administered borneol (50 mg/kg body weight (BW)) or 0.5% carboxymethyl-cellulose sodium for 7 consecutive days, and then intragastrically administered florfenicol (25 mg/kg BW) on the eighth day. Pharmacokinetic studies showed that borneol significantly decreased the area under the concentration-time curve from zero to infinity (AUC(0-t)), time to reach peak concentration (Tmax), and the peak concentration (Cmax) values of florfenicol, whereas the values of mean residence time from zero to infinity (MRT(0-t)), elimination half-life (t1/2z), apparent volume of distribution fraction of the dose absorbed (Vz), and plasma clearance fraction of the dose absorbed (CLz) were increased significantly. Furthermore, the mRNA expression levels of multidrug resistance 1 (MDR1) and cytochrome P450 3A1 (CYP3A1) in the jejunum and of CYP1A2 and CYP2C11 in the liver were significantly upregulated by borneol. In conclusion, borneol decreased absorption, increased clearance, improved distribution, and increased the mean residence time of florfenicol in rats, possibly through regulating the mRNA expression levels of drug-metabolizing enzymes and efflux transporters.


Assuntos
Hidrocarboneto de Aril Hidroxilases , Citocromo P-450 CYP1A2 , Animais , Canfanos , Sistema Enzimático do Citocromo P-450 , Família 2 do Citocromo P450 , Resistência a Múltiplos Medicamentos , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Esteroide 16-alfa-Hidroxilase , Tianfenicol/análogos & derivados
4.
Xenobiotica ; 51(9): 961-967, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29160125

RESUMO

Paeoniflorin is the major constituent in extracts of the paeony root, the purpose of the present study was to assess the effects of paeoniflorin on the activities and mRNA expression of the rat hepatic drug-metabolizing enzymes cytochrome P450 (CYP1A2), CYP2C11 and CYP3A1 in vivo.Sprague-Dawley (SD) male rats were treated with paeoniflorin at the dosage of 25, 50 and 100 mg/kg or 0.9% sodium chloride solution by intragastric administration for 7 days, then were given probe drugs phenacetin (CYP1A2), tolbutamide (CYP2C11), or midazolam (CYP3A1) orally on the eighth day. Blood samples were collected at various times, and the plasma concentrations of the probe drugs were estimated with ultra-high-performance liquid chromatography. The mRNA expression levels of rat hepatic CYP1A2, CYP2C11 and CYP3A1 were analysed with real-time PCR.The pharmacokinetic results indicated that paeoniflorin inhibits the activities of CYP1A2, CYP2C11 and CYP3A1 in vivo. The effect was most pronounced on CYP3A1, according to the United States Food and Drug Administration classification of inhibitors of CYP3A, it reached the category of moderate inhibition. The mRNA expression levels of 3 CYP enzymes were also tended to be inhibited.We conclude that paeoniflorin can inhibit the activities of CYP1A2, CYP2C11 and CYP3A1 in vivo, which may affect the metabolism of drugs that are primarily dependent on these pathways.


Assuntos
Hidrocarboneto de Aril Hidroxilases , Citocromo P-450 CYP1A2 , Animais , Citocromo P-450 CYP1A2/genética , Família 2 do Citocromo P450/genética , Glucosídeos/farmacologia , Masculino , Monoterpenos , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Esteroide 16-alfa-Hidroxilase/genética
5.
J Ethnopharmacol ; 264: 113354, 2021 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-32898626

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Geissoschizine methyl ether (GM), an indole alkaloid from Uncaria hook, is an active ingredient in the traditional Japanese Kampo medicine yokukansan, which is used to treat neurosis, insomnia, irritability, and night crying in children. AIM OF THE STUDY: Recent our pharmacokinetic studies suggested that there may be gender differences in the plasma concentrations of GM in rats, but not in humans. However, the details of this difference remain unverified. The purpose of this study was to clarify the reasons for the gender differences in rats. MATERIALS AND METHODS: GM plasma pharmacokinetics was compared in male and female rats orally administered yokukansan (4 g/kg). To confirm the involvement of cytochrome P450 (CYP) in GM liver metabolism, GM was incubated with male and female rat liver S9 fraction in the absence or presence of 1-aminobenzotriazole (a nonspecific CYP inhibitor). CYP isoforms involved in GM metabolism were estimated using recombinant rat CYP isoforms and anti-rat CYP antibodies. RESULTS: The maximum GM plasma concentrations were significantly higher in female than in male rats. When GM was incubated with rat liver S9 fractions, GM reduction was more striking in male S9 (69.3%) than that in female S9 (10.0%) and was completely blocked with nonspecific CYP inhibitor 1-aminobenzotriazole. Screening experiments using recombinant rat cytochrome P450 (CYP) isoforms showed that CYP1A1, CYP2C6, CYP2C11, CYP2D1, and CYP3A2 were involved in GM metabolism. Of these CYP isoforms, the use of anti-rat CYP antibodies indicated that male-dependent CYP2C11 and CYP3A2 were predominantly involved in the liver microsomal GM metabolism with gender differences. CONCLUSIONS: These results suggest that the cause of gender differences in plasma GM pharmacokinetics in rats is most likely because of male-dependent CYP2C11 and CYP3A2, and provide also useful information to further evaluate the pharmacological and toxicological effects in future. This study is the first to demonstrate that the gender differences in plasma GM pharmacokinetics in rats are caused by the gender-dependent metabolism of GM.


Assuntos
Alcaloides Indólicos/sangue , Microssomos Hepáticos/efeitos dos fármacos , Caracteres Sexuais , Uncaria , Animais , Hidrocarboneto de Aril Hidroxilases/metabolismo , Citocromo P-450 CYP3A/metabolismo , Família 2 do Citocromo P450/metabolismo , Medicamentos de Ervas Chinesas/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Feminino , Alcaloides Indólicos/metabolismo , Alcaloides Indólicos/farmacologia , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , Microssomos Hepáticos/enzimologia , Plasma/efeitos dos fármacos , Plasma/metabolismo , Ratos , Ratos Sprague-Dawley , Esteroide 16-alfa-Hidroxilase/metabolismo
6.
Brain Res Bull ; 163: 57-64, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32707261

RESUMO

Cytochrome P450 (CYP) epoxygenases have been considered the main producers of epoxyeicosatrienoic acids (EETs) through the oxidation of arachidonic acid (AA). EETs display various biological properties, notably their powerful anti-inflammatory activities. In the brain, EETs have proven to be neuroprotective and to improve neuroinflammation. However, it is known that inflammation could modify CYP expression. We have previously reported that an inflammatory process in astrocytes is able to down-regulate CYP2J3 and CYP2C11 mRNA, protein levels, and activity (Navarro-Mabarak et al., 2019). In this work, we evaluated the effect of neuroinflammation in protein expression of CYP epoxygenases in the brain. Neuroinflammation was induced by the intraperitoneal administration of LPS (1 mg/kg) to male Wistar rats and was corroborated by IL-6, GFAP, and Iba-1 protein levels in the cortex over time. CYP2J3 and CYP2C11 protein levels were also evaluated in the cortex after 6, 12, 24, 48, and 72 h of LPS treatment. Our results show for the first time that neuroinflammation is able to downregulate CYP2J3 and CYP2C11 protein expression in the brain cortex.


Assuntos
Hidrocarboneto de Aril Hidroxilases/metabolismo , Encéfalo/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Família 2 do Citocromo P450/metabolismo , Regulação para Baixo/fisiologia , Mediadores da Inflamação/metabolismo , Esteroide 16-alfa-Hidroxilase/metabolismo , Animais , Hidrocarboneto de Aril Hidroxilases/antagonistas & inibidores , Encéfalo/efeitos dos fármacos , Família 2 do Citocromo P450/antagonistas & inibidores , Regulação para Baixo/efeitos dos fármacos , Lipopolissacarídeos/toxicidade , Masculino , Ratos , Ratos Wistar , Esteroide 16-alfa-Hidroxilase/antagonistas & inibidores
7.
J Pharm Pharmacol ; 72(7): 938-955, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32307724

RESUMO

OBJECTIVES: N-(2-hydroxyphenyl)-2-propylpentanamide (HO-AAVPA), a derivative of valproic acid (VPA), has been proposed as a potential anticancer agent due to its improved antiproliferative effects in some cancer cell lines. Although there is evidence that VPA is metabolized by cytochrome P450 2C11 rat isoform, HO-AAVPA CYP-mediated metabolism has not yet been fully explored. Therefore, in this work, the biotransformation of HO-AAVPA by CYP2C11 was investigated. METHODS: Kinetic parameters and spectral interaction between HO-AAVPA and CYP were evaluated using rat liver microsomes. The participation of CYP2C11 in metabolism of HO-AAVPA was confirmed by cimetidine (CIM) inhibition assay. Docking and molecular dynamics simulations coupled to MMGBSA methods were used in theoretical study. KEY FINDINGS: HO-AAVPA is metabolized by CYP enzymes (KM  = 38.94 µm), yielding a hydroxylated metabolite according to its HPLC retention time (5.4 min) and MS analysis (252.2 m/z). In addition, CIM inhibition in rat liver microsomes (Ki  = 59.23 µm) confirmed that CYP2C11 is mainly involved in HO-AAVPA metabolism. Furthermore, HO-AAVPA interacts with CYP2C11 as a type I ligand. HO-AAVPA is stabilized at the CYP2C11 ligand recognition site through a map of interactions similar to other typical CYP2C11 substrates. CONCLUSION: Therefore, rat liver CYP2C11 isoform is able to metabolize HO-AAVPA.


Assuntos
Amidas/farmacocinética , Hidrocarboneto de Aril Hidroxilases/metabolismo , Biotransformação , Família 2 do Citocromo P450/metabolismo , Microssomos Hepáticos , Pentanos/farmacocinética , Esteroide 16-alfa-Hidroxilase/metabolismo , Animais , Antineoplásicos/farmacocinética , Proliferação de Células/efeitos dos fármacos , Estabilidade de Medicamentos , Hidroxilação , Isoenzimas/metabolismo , Oxigenases de Função Mista/metabolismo , Simulação de Acoplamento Molecular , Ratos , Ácido Valproico/farmacologia
8.
Molecules ; 24(23)2019 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-31775347

RESUMO

The inhibitory effect of new chemical entities on rat liver P450 marker activities was investigated in a functional approach towards drug development. Treatment of colorectal cancer (CRC) and chemoprevention using salicylic acid has gained a lot of attention, mainly in the prevention of the onset of colon cancer. Thus, an in vitro inhibitory effect of salicylic acid on rat CYP2C11 activity was examined by using high performance liquid chromatography (HPLC). High performance liquid chromatography analysis of a CYP2C11 assay was developed on a reversed phase C18 column (SUPELCO 25 cm × 4.6 mm × 5 µm) at 243 nm using 32% phosphate buffer (pH 3.36) and 68% methanol as a mobile phase. The CYP2C11 assay showed good linearity for all components (R2 > 0.999). Substrates and metabolites were found to be stable for up to 72 hours. Additionally, the method demonstrated good reproducibility, intra- and inter-day precision (<15%), acceptable recovery and accuracy (80%-120%), and low detection (1.3501 µM and 3.2757 µM) and quantitation limit values (4.914 µM and 9.927 µM) for 16α-hydroxytestosterone and testosterone, respectively. Salicylic acid acts reversibly as a noncompetitive (weak) inhibitor with Ki = 84.582 ± 2.67 µM (concentration of inhibitor to cause 50% inhibition of original enzyme activity (IC50) = 82.70 ± 2.67 µM) for CYP2C11 enzyme activity. This indicates a low potential to cause toxicity and drug-drug interactions.


Assuntos
Hidrocarboneto de Aril Hidroxilases/antagonistas & inibidores , Inibidores das Enzimas do Citocromo P-450/farmacologia , Família 2 do Citocromo P450/antagonistas & inibidores , Fígado/efeitos dos fármacos , Ácido Salicílico/farmacologia , Esteroide 16-alfa-Hidroxilase/antagonistas & inibidores , Animais , Hidrocarboneto de Aril Hidroxilases/química , Catálise , Cromatografia Líquida de Alta Pressão , Inibidores das Enzimas do Citocromo P-450/química , Família 2 do Citocromo P450/química , Desenvolvimento de Medicamentos , Humanos , Fígado/enzimologia , Ratos , Ácido Salicílico/química , Esteroide 16-alfa-Hidroxilase/química
9.
Neurochem Int ; 129: 104499, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31271766

RESUMO

Cytochrome P450 (CYP) epoxygenases and their metabolic products, epoxyeicosatrienoic acids (EETs), have been proposed as important therapeutic targets in the brain. However, CYP expression can be modified by the presence of diverse pro-inflammatory cytokines and the subsequent activation of the NF-κB pathway. It has been indicated that CYP epoxygenases are down-regulated by inflammation in the heart, kidney and liver. However, up to this point, there has been no evidence regarding regulation of CYP epoxygenases during inflammation in the brain. Therefore, in order to explore the effects of inflammation and NF-κB activation in CYP2J3 and CYP2C11 regulation, rat primary astrocytes cultures were treated with LPS with and without IMD-0354 (selective NF-κB inhibitor). Cyp2j3 and Cyp2c11 mRNA expression was determined by qRT-PCR; protein expression was determined by immunofluorescence and by Western Blot and total epoxygenase activity was determined by the quantification of EETs by ELISA. NF-κB binding sites in Cyp2j3 and Cyp2c11 promoter regions were bioinformatically predicted and Electrophoretic Mobility Shift Assays (EMSA) were performed to determine if each hypothetic response element was able to bind NF-κB complexes. Results shown that LPS treatment is able to down-regulate astrocyte CYP2J3 and CYP2C11 mRNA, protein and activity. Additionally, we have identified NK-κB as the transcription factor involved in this regulation.


Assuntos
Astrócitos/metabolismo , Regulação da Expressão Gênica , Inflamação/metabolismo , NF-kappa B/fisiologia , Animais , Hidrocarboneto de Aril Hidroxilases , Benzamidas/farmacologia , Células Cultivadas , Córtex Cerebral/citologia , Sistema Enzimático do Citocromo P-450 , Família 2 do Citocromo P450 , Regulação para Baixo/efeitos dos fármacos , Eicosanoides/biossíntese , Endotoxinas/farmacologia , Inflamação/induzido quimicamente , Inflamação/genética , Masculino , NF-kappa B/antagonistas & inibidores , Cultura Primária de Células , Regiões Promotoras Genéticas , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Wistar , Esteroide 16-alfa-Hidroxilase , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/genética
10.
Biopharm Drug Dispos ; 40(7): 225-233, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31215040

RESUMO

The study examined the effect of doxorubicin (DOX) on the hepatic expression of CYP2C and its activity for metabolizing tolbutamide (TB), a specific CYP2C substrate, in rats and whether the pharmacokinetics of tolbutamide were altered by doxorubicin exposure. The expression level of hepatic CYP2C11 was depressed 1 day after doxorubicin administration (day 1), and this effect on CYP2C11 was augmented on day 4. However, the expression level of hepatic CYP2C6 remained unchanged. The activity of tolbutamide 4-hydroxylation in hepatic microsomes was decreased with time following doxorubicin administration. Regarding the enzyme kinetic parameters for tolbutamide 4-hydroxylation on day 4, the maximum velocity (Vmax ) was significantly lower in the DOX group than that in the control group, while the Michaelis constant (Km ) was unaffected. On pharmacokinetic examination, the total clearance (CLtot ) of tolbutamide on day 4 was increased, despite the decreased metabolic capacity. On the other hand, the serum unbound fraction (fu ) of tolbutamide was elevated with a reduced serum albumin concentration in the DOX group. Contrary to CLtot , CLtot /fu , a parameter approximated to the hepatic intrinsic clearance of unbound tolbutamide, was estimated to be significantly reduced in the DOX group. These findings indicate that the metabolic capacity of CYP2C11 in the liver is depressed time-dependently by down-regulation after doxorubicin exposure in rats, and that the decreased enzyme activity of TB 4-hydroxylation in hepatic microsomes reflects the pharmacokinetic change of unbound tolbutamide, not total tolbutamide, in serum.


Assuntos
Antibióticos Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Hipoglicemiantes/farmacocinética , Tolbutamida/farmacocinética , Animais , Hidrocarboneto de Aril Hidroxilases/metabolismo , Família 2 do Citocromo P450/metabolismo , Interações Medicamentosas , Hidroxilação/efeitos dos fármacos , Hipoglicemiantes/sangue , Masculino , Taxa de Depuração Metabólica/efeitos dos fármacos , Microssomos Hepáticos/metabolismo , Ratos Sprague-Dawley , Albumina Sérica/metabolismo , Esteroide 16-alfa-Hidroxilase/metabolismo , Tolbutamida/sangue
11.
J Pharm Biomed Anal ; 174: 367-375, 2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31202879

RESUMO

Fufang Danshen Dripping Pill (FDDP) and Clopidogrel Bisulfate Tablet (CBT) are usually combined for treatment of coronary artery diseases in clinical. To investigate the pharmacokinetic interaction between FDDP and CBT after oral administration of FDDP, CBT and their combination in rats, a novel LC-MS method with segmented scan modes (multiple reaction monitoring and selected ion monitoring) and polarity (positive and negative ionization) was developed. Clopidogrel and the main active ingredients of FDDP, with different chemical and ionization properties, were simultaneously quantified in plasma in a single run. The method was validated in terms of specificity, linearity, precision, accuracy, recovery, matrix effect and stability. As a result, co-administration of FDDP and CBT significantly altered the pharmacokinetic parameters of danshensu, ginsenoside Rb1, dihydrotanshinone I, tanshinone I and tanshinone IIA of FDDP, as well as clopidogrel. Mechanism studies suggested that induction of liver cytochrome P450 isozymes CYP2C11 and CYP3A1 by co-administration, as well as inhibition of carboxyl esterase 1, was partly responsible for FDDP-CBT pharmacokinetic interactions. The developed LC-MS method could be used to simultaneously quantify different types of in vivo analytes in a single run, and the results could be used for clinical medication guidance of FDDP and CBT.


Assuntos
Clopidogrel/farmacocinética , Medicamentos de Ervas Chinesas/farmacocinética , Abietanos/farmacocinética , Administração Oral , Animais , Hidrocarboneto de Aril Hidroxilases/metabolismo , Canfanos , Cromatografia Líquida , Citocromo P-450 CYP3A/metabolismo , Família 2 do Citocromo P450/metabolismo , Ginsenosídeos/farmacocinética , Lactatos/farmacocinética , Modelos Lineares , Masculino , Panax notoginseng , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Salvia miltiorrhiza , Esteroide 16-alfa-Hidroxilase/metabolismo , Espectrometria de Massas em Tandem
12.
Eur J Drug Metab Pharmacokinet ; 44(6): 787-796, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31175627

RESUMO

BACKGROUND AND OBJECTIVES: Honokiol, a major constituent isolated from Magnolia officinalis, is regarded as a phytochemical marker and bioactive substance present in many traditional Chinese medicines. However, the effect of honokiol on cytochrome P450 (CYP) has not been thoroughly investigated. The aim of this study was to investigate the effect of honokiol on CYP1A2 and CYP2C11 in vitro and in vivo. METHODS: The effect of honokiol on CYP1A2 and CYP2C11 was investigated with rat liver microsomes (RLMs) by measuring phenacetin and tolbutamide metabolism (probe drugs for CYP1A2 and CYP2C11, respectively), and then explored in vivo by measuring the effect of honokiol (2.5 and 5 mg/kg, intravenous injection) on the pharmacokinetics of theophylline and tolbutamide (probe drugs for CYP1A2 and CYP2C11, respectively) in rats in vivo. RESULTS: Honokiol inhibited the formation of acetaminophen from phenacetin and 4-hydroxytolbutamide from tolbutamide in RLMs, with inhibition constant (Ki) values of 1.6 µM and 16.5 µM, respectively. In vivo, honokiol (2.5 or 5.0 mg/kg) increased the half-life (t1/2) of theophylline by 40.9% and 119.9%, decreased the clearance (CL) by 23.8% and 42.9%, and increased the area under the curve (AUC) by 41.3% and 83.4%, respectively. Similarly, the t1/2 of tolbutamide increased by 25.5% and 33.8%, the CL decreased by 14.3% and 19.1%, and the AUC increased by 19.2% and 25.7%, respectively. CONCLUSION: The inhibition of CYP1A2 by honokiol is greater than the inhibition of CYP2C11. The changes in the pharmacokinetics of theophylline and tolbutamide in rats treated with honokiol are due to the inhibition of CYP1A2 and CYP2C11 activity in a dose-dependent manner.


Assuntos
Hidrocarboneto de Aril Hidroxilases/antagonistas & inibidores , Compostos de Bifenilo/farmacologia , Família 2 do Citocromo P450/antagonistas & inibidores , Lignanas/farmacologia , Esteroide 16-alfa-Hidroxilase/antagonistas & inibidores , Animais , Compostos de Bifenilo/química , Compostos de Bifenilo/farmacocinética , Citocromo P-450 CYP1A2 , Citocromos/antagonistas & inibidores , Lignanas/química , Lignanas/farmacocinética , Masculino , Ratos , Ratos Sprague-Dawley , Teofilina/farmacocinética , Tolbutamida/farmacocinética
13.
Epilepsy Res ; 153: 14-18, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30927680

RESUMO

Dietary supplements, herbal medicines, and other foods may affect the pharmacokinetics and/or pharmacodynamics of carbamazepine (CBZ), which may possibly lead to potential drug-drug/herb-drug interactions, as CBZ has a narrow therapeutic window. Sinapic acid (SA) is a bioactive phytoconstituent used as a dietary supplement for the treatment of epilepsy. This study determined the effects of SA on the pharmacokinetics of CBZ and proposed a possible interaction mechanism in twenty-four male wistar rats (180-210 g). A single CBZ dose (80 mg/kg) was administered orally to rats with or without SA pretreatment (20 mg/kg p.o. per day for 7 days, n = 6). The CBZ concentration in plasma samples was determined by using a sensitive reversed-phase high-performance liquid chromatography assay. The pharmacokinetic parameters were calculated by using non-compartmental analysis. Significance was determined through Dunnett's multiple comparison test or one-way analysis of variance as appropriate; p < 0.05 were considered significant. The change in the pharmacokinetic parameters (Cmax, Tmax, AUC0-t, AUC0-∞, T½, and kel) of CBZ was evaluated after the administration of CBZ alone or after CBZ co-administration with SA pretreatment. The plasma concentration of CBZ was higher after SA pretreatment than that without pretreatment. The pharmacokinetics of orally administered CBZ were found to be significantly altered (p < 0.05) in rats pretreated with SA compared to those in rats administered CBZ alone. The increases in the Cmax, AUC0-t, T1/2, and MRT of CBZ were 29.79%, 57.18%, 77.18%, and 58.31%, respectively, whereas the kel and apparent oral CL/F were significantly reduced (p < 0.05) in rats pretreated with SA compared to those in rats not pretreated with SA (43.87% and 42.50%, respectively). However, no significant change was observed in the Tmax of CBZ in rats pretreated with SA compared to that in rats that did not receive pretreatment. The enhancement in Cmax, AUC0-t, T1/2, and MRT and the reduction in Kel and CL/F values resulted from the significant inhibition of CYP3 A2, the CYP2C11-mediated metabolism of CBZ in the liver, and the inhibition of intestinal P-glycoprotein/MDR1, which enhanced the rate of CBZ absorption. Further studies are required to determine the clinical relevance of these observations.


Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Anticonvulsivantes/administração & dosagem , Hidrocarboneto de Aril Hidroxilases/metabolismo , Carbamazepina/administração & dosagem , Ácidos Cumáricos/farmacocinética , Citocromo P-450 CYP3A/metabolismo , Família 2 do Citocromo P450/metabolismo , Indicadores e Reagentes/farmacocinética , Mucosa Intestinal/efeitos dos fármacos , Esteroide 16-alfa-Hidroxilase/metabolismo , Administração Oral , Animais , Área Sob a Curva , Interações Medicamentosas , Mucosa Intestinal/metabolismo , Intestinos/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar
14.
Xenobiotica ; 49(12): 1478-1484, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30724651

RESUMO

1. CYP2C11 is the most abundant isoform of cytochrome P450s (CYPs) in male rats and is considered the main enzyme for warfarin metabolism. 2. To further access the in vivo function of CYP2C11 in warfarin metabolism and efficacy, a CYP2C11-null rat model was used to study warfarin metabolism with both in vitro and in vivo approaches. Prothrombin time (PT) of warfarin was also determined. 3. The maximum rate of metabolism (Vmax) and intrinsic clearance (CLint) of liver microsomes from CYP2C11-null males were reduced by 37 and 64%, respectively, compared to those in Sprague Dawley (S-D) rats. The Km of liver microsomes from CYP2C11-null males was increased by 73% compared to that of S-D rats. The time to reach the maximum plasma concentration (Tmax) of warfarin in CYP2C11-null males was significantly delayed compared to that in S-D males, and the CL rate was also reduced. The PT of CYP2C11-null rats was moderately longer than that of S-D rats. 4. In conclusion, the clearance rate of warfarin was mildly decreased and its anticoagulant effect was moderately increased in male rats following CYP2C11 gene knockout. CYP2C11 played a certain role in the clearance and efficacy of warfarin, while it did not seem to be essential.


Assuntos
Hidrocarboneto de Aril Hidroxilases/genética , Família 2 do Citocromo P450/genética , Esteroide 16-alfa-Hidroxilase/genética , Varfarina/farmacocinética , Animais , Anticoagulantes/farmacocinética , Hidrocarboneto de Aril Hidroxilases/metabolismo , Família 2 do Citocromo P450/metabolismo , Feminino , Técnicas de Inativação de Genes , Masculino , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/metabolismo , Ratos Sprague-Dawley , Esteroide 16-alfa-Hidroxilase/metabolismo
15.
Cancer Res ; 78(16): 4716-4730, 2018 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-29921690

RESUMO

Primary prostate cancer can have extensive microheterogeneity, but its contribution to the later emergence of metastatic castration-resistant prostate cancer (mCRPC) remains unclear. In this study, we microdissected residual prostate cancer foci in radical prostatectomies from 18 men treated with neoadjuvant-intensive androgen deprivation therapy (leuprolide, abiraterone acetate, and prednisone) and analyzed them for resistance mechanisms. Transcriptome profiling showed reduced but persistent androgen receptor (AR) activity in residual tumors, with no increase in neuroendocrine differentiation. Proliferation correlated negatively with AR activity but positively with decreased RB1 expression, and whole-exome sequencing (WES) further showed enrichment for RB1 genomic loss. In 15 cases where 2 or 3 tumor foci were microdissected, WES confirmed a common clonal origin but identified multiple oncogenic alterations unique to each focus. These findings show that subclones with oncogenic alterations found in mCRPC are present in primary prostate cancer and are selected for by neoadjuvant-intense androgen deprivation therapy. In particular, this study indicates that subclonal RB1 loss may be more common than previously appreciated in intermediate- to high-risk primary prostate cancer and may be an early event, independent of neuroendocrine differentiation, in the development of mCRPC. Comprehensive molecular analyses of primary prostate cancer may detect aggressive subclones and possibly inform adjuvant strategies to prevent recurrence.Significance: Neoadjuvant androgen deprivation therapy for prostate cancer selects for tumor foci with subclonal genomic alterations, which may comprise the origin of metastatic castration-resistant prostate cancer. Cancer Res; 78(16); 4716-30. ©2018 AACR.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Resistencia a Medicamentos Antineoplásicos/genética , Recidiva Local de Neoplasia/tratamento farmacológico , Neoplasias de Próstata Resistentes à Castração/tratamento farmacológico , Acetato de Abiraterona/administração & dosagem , Idoso , Antagonistas de Androgênios/administração & dosagem , Hidrocarboneto de Aril Hidroxilases/genética , Biomarcadores Tumorais , Carcinogênese , Evolução Clonal , Família 2 do Citocromo P450/genética , Humanos , Masculino , Pessoa de Meia-Idade , Terapia Neoadjuvante , Recidiva Local de Neoplasia/genética , Recidiva Local de Neoplasia/patologia , Prostatectomia , Neoplasias de Próstata Resistentes à Castração/genética , Neoplasias de Próstata Resistentes à Castração/patologia , Receptores Androgênicos/genética , Esteroide 16-alfa-Hidroxilase/genética
16.
Plant Physiol Biochem ; 131: 70-77, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29735370

RESUMO

Potato (Solanum tuberosum) is a major food crop, while the most tissues of potato accumulates steroidal glycoalkaloids (SGAs) α-solanine and α-chaconine. Since SGAs confer a bitter taste on human and show the toxicity against various organisms, reducing the SGA content in the tubers is requisite for potato breeding. However, generation of SGA-free potato has not been achieved yet, although silencing of several SGA biosynthetic genes led a decrease in SGAs. Here, we show that the knockout of St16DOX encoding a steroid 16α-hydroxylase in SGA biosynthesis causes the complete abolition of the SGA accumulation in potato hairy roots. Nine candidate guide RNA (gRNA) target sequences were selected from St16DOX by in silico analysis, and the two or three gRNAs were introduced into a CRISPR/Cas9 vector designated as pMgP237-2A-GFP that can express multiplex gRNAs based on the pre-tRNA processing system. To establish rapid screening of the candidate gRNAs that can efficiently mutate the St16DOX gene, we used a potato hairy root culture system for the introduction of the pMgP237 vectors. Among the transgenic hairy roots, two independent lines showed no detectable SGAs but accumulated the glycosides of 22,26-dihydroxycholesterol, which is the substrate of St16DOX. Analysis of the two lines with sequencing exhibited the mutated sequences of St16DOX with no wild-type sequences. Thus, generation of SGA-free hairy roots of tetraploid potato was achieved by the combination of the hairy root culture and the pMgP237-2A-GFP vector. This experimental system is useful to evaluate the efficacy of candidate gRNA target sequences in the short-term.


Assuntos
Sistemas CRISPR-Cas/genética , Edição de Genes/métodos , Genes de Plantas/genética , Raízes de Plantas/genética , Solanina/metabolismo , Solanum tuberosum/genética , Esteroide 16-alfa-Hidroxilase/genética , Técnicas de Inativação de Genes/métodos , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Análise de Sequência de DNA , Solanum tuberosum/metabolismo , Esteroide 16-alfa-Hidroxilase/metabolismo
17.
Food Chem Toxicol ; 116(Pt B): 369-378, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29698782

RESUMO

The aim of this study is to assess a potential mechanism by which the serotonergic system can control the expression and activity of cytochrome (CYP) 2C11 and CYP3A isoforms during liver insufficiency. A rat model of diethylnitrosamine (DEN)-induced liver insufficiency was developed by administering 50 mg/kg of DEN twice a week for 7 weeks. Dysfunction of the serotonergic system was evoked by feeding the rats with a tryptophan-free diet for three weeks. Dysfunction of the serotonergic system during liver insufficiency decreased the level of proinflammatory cytokines (TGF-ß and IL-1ß) and increased the level of an anti-inflammatory cytokine (IL-4). Simultaneously, activation of the repressive mechanism IL-4/JAK1/STAT6/SOCS1 of the JAK2/STAT5b-mediated signal transduction pathway and the pERK1/2/GR/STAT6 signal transduction pathway resulted in the suppression of the CYP2C11 and CYP3A isoforms. Moreover, dysfunction of the serotonergic system during liver insufficiency equalized the level of testosterone to the basal level, did not change the steady state of the corticosterone level and significantly enhanced the reduced level of growth hormone. An altered cytokine profile under control of the serotonergic system determines the regulation of CYP2C11 and CYP3A isoforms during liver insufficiency through mechanisms based on posttranscriptional and posttranslational processes.


Assuntos
Hidrocarboneto de Aril Hidroxilases/metabolismo , Citocromo P-450 CYP3A/metabolismo , Família 2 do Citocromo P450/metabolismo , Citocinas/sangue , Insuficiência Hepática/enzimologia , Serotonina/fisiologia , Esteroide 16-alfa-Hidroxilase/metabolismo , Animais , Biomarcadores/metabolismo , Peso Corporal , Dietilnitrosamina/toxicidade , Modelos Animais de Doenças , Insuficiência Hepática/induzido quimicamente , Insuficiência Hepática/metabolismo , Mediadores da Inflamação/metabolismo , Interleucina-1beta/metabolismo , Interleucina-4/metabolismo , Fígado/metabolismo , Fígado/patologia , Masculino , Tamanho do Órgão , Processamento de Proteína Pós-Traducional , Processamento Pós-Transcricional do RNA , Ratos Wistar , Transdução de Sinais , Testosterona/metabolismo , Fator de Crescimento Transformador beta/metabolismo
18.
Drug Metab Dispos ; 46(6): 786-793, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29555828

RESUMO

Our recent work suggested a negative effect for the serotonergic innervation of the paraventricular nuclei (PVN) of the hypothalamus on growth hormone secretion and growth hormone-dependent expression of CYP2C11. The aim of our present research was to determine the effect of the activation of the 5-hydroxytryptamine [(5-HT) serotonin] 5-HT1 or 5-HT2 receptors in the PVN on the expression and activity of cytochrome P450 in male rat liver. The serotonergic agonists 5-carboxyamidotryptamine [(5-CT), a 5-HT1 receptor-type agonist], 8-hydroxy-2-(di-n-propyloamino)-tetralin [(8-OH-DPAT), a 5-HT1A receptor agonist], sumatriptan (a 5-HT1B/D receptor agonist), and 2,5-dimethoxy-4-iodoamphetamine [(DOI), a 5-HT2A/C receptor agonist] were individually injected into the PVN. The liver cytochrome P450 activity and expression and the levels of serum and pituitary and hypothalamic hormones were measured. 5-CT and 8-OH-DPAT significantly decreased the activity and expression of CYP2C11 at both the mRNA and protein levels, which was accompanied by an increase in pituitary and hypothalamic somatostatin levels and a decrease in the serum growth hormone concentration. The expression of CYP3A1/23 also decreased. The serum corticosterone concentration declined after the injection of 8-OH-DPAT. The obtained results indicated that 5-HT1A but not the 5-HT1B/D or 5-HT2 receptors in the PVN are engaged in the negative neuroendocrine regulation of cytochrome P450 via the stimulation of hypothalamic somatostatin secretion and in the decreases in the serum growth hormone and corticosterone concentrations. Since the affected enzymes metabolize steroids and drugs and 5-HT1A receptors are engaged in the action of psychotropic drugs, the results obtained may be of both physiologic and pharmacological meaning.


Assuntos
Hidrocarboneto de Aril Hidroxilases/metabolismo , Família 2 do Citocromo P450/metabolismo , Núcleo Hipotalâmico Paraventricular/metabolismo , Receptor 5-HT1A de Serotonina/metabolismo , Esteroide 16-alfa-Hidroxilase/metabolismo , 8-Hidroxi-2-(di-n-propilamino)tetralina/farmacologia , Animais , Corticosterona/metabolismo , Hormônio do Crescimento/metabolismo , Fígado , Masculino , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Ratos , Ratos Wistar , Serotonina/metabolismo , Agonistas do Receptor de Serotonina/farmacologia
19.
Drug Metab Dispos ; 46(5): 525-531, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29444903

RESUMO

CYP2C11 is involved in the metabolism of many drugs in rats. To assess the roles of CYP2C11 in physiology and drug metabolism, a CYP2C11-null rat model was generated using the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9method. A 2-base pair insertion was added to exon 6 of CYP2C11 in Sprague-Dawley rats. CYP2C11 was not detected by western blotting in liver microsomes of CYP2C11-null rats. No off-target effects were found at 11 predicted sites of the knockout model. The CYP2C11-null rats were viable and had no obvious abnormalities, with the exception of reduced fertility. Puberty in CYP2C11-null rats appeared to be delayed by ∼20 days, and the average litter size fell by 43%. Tolbutamide was used as a probe in this drug metabolism study. In the liver microsomes of CYP2C11-null rats, the Vmax and intrinsicclearance values decreased by 22% and 47%, respectively, compared with those of wild-type rats. The Km values increased by 47% compared with that of wild types. However, our pharmacokinetics study showed no major differences in any parameters between the two strains, in both males and females. In conclusion, a CYP2C11-null rat model was successfully generated and is a valuable tool to study the in vivo function of CYP2C11.


Assuntos
Hidrocarboneto de Aril Hidroxilases/genética , Sistemas CRISPR-Cas/genética , Família 2 do Citocromo P450/genética , Esteroide 16-alfa-Hidroxilase/genética , Animais , Sistemas CRISPR-Cas/efeitos dos fármacos , Feminino , Inativação Metabólica/fisiologia , Masculino , Microssomos Hepáticos/efeitos dos fármacos , Modelos Animais , Ratos , Ratos Sprague-Dawley , Tolbutamida/farmacologia
20.
Xenobiotica ; 48(9): 911-919, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29052493

RESUMO

1. Aspirin (ASA) and clopidogrel (CLP) are used in combination as dual antiplatelet therapy (DAPT) for acute coronary syndrome based on their complementary mechanisms for platelet aggregation inhibition. However, the pharmacokinetics of such drug combination usage has not been thoroughly investigated. 2. In the current study, an LC-MS/MS method was developed to simultaneously determine the plasma concentrations of ASA and its metabolite salicylic acid (SA) with CLP and its metabolites, clopidogrel carboxylic acid (CLPM) and clopidogrel active metabolite derivative (CAMD). The pharmacokinetics of ASA, SA, CLP, CLPM and CAMD in rats receiving two-week DAPT with ASA and CLP were then determined. 3. After two-week DAPT with ASA and CLP in rats, the activities of aspirin esterase and rCyp2c11, enzymes mediating rat metabolism of ASA and CLP, respectively, in prepared rat liver microsomes were measured followed by further determination of rCyp2c11 mRNA expressions. The results demonstrated that DAPT led to minimal impact on aspirin esterase activity but significant decrease in rCyp2c11 activity and mRNA expression. 4. In conclusion, our findings on impairment in rCyp2C11 activity and mRNA expression by DAPT in rats could provide guidance on its safe clinical use with other CYP 2C19 substrates.


Assuntos
Hidrocarboneto de Aril Hidroxilases/metabolismo , Aspirina/farmacocinética , Família 2 do Citocromo P450/metabolismo , Fígado/efeitos dos fármacos , Inibidores da Agregação Plaquetária/farmacologia , Esteroide 16-alfa-Hidroxilase/metabolismo , Ticlopidina/análogos & derivados , Animais , Hidrocarboneto de Aril Hidroxilases/genética , Aspirina/sangue , Cromatografia Líquida/métodos , Clopidogrel , Família 2 do Citocromo P450/genética , Inativação Metabólica , Fígado/metabolismo , Masculino , Inibidores da Agregação Plaquetária/farmacocinética , Ratos Sprague-Dawley , Ácido Salicílico/metabolismo , Esteroide 16-alfa-Hidroxilase/genética , Espectrometria de Massas em Tandem/métodos , Ticlopidina/sangue , Ticlopidina/farmacocinética
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