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Chemical Research in Toxicology Apr 2021Binimetinib is a selective MEK1/2 inhibitor, which is indicative of melanoma. We aimed to investigate the inhibitory effect of binimetinib on cytochrome P450 using human...
Binimetinib is a selective MEK1/2 inhibitor, which is indicative of melanoma. We aimed to investigate the inhibitory effect of binimetinib on cytochrome P450 using human liver microsomes. Binimetinib was demonstrated to display reversible and time-dependent inhibitory effects on human CYP1A2. Binimetinib can inhibit the activity of phenacetin deethylation with IC of 5.6 μM. A 30 min preincubation of binimetinib with NADPH-supplemented human liver microsomes raised a significant left IC shift (6.5-fold), from 5.69-0.88 μM. The inactivation parameters and were 0.063 min and 15.47 μM, and the half-life of inactivation was 11 min. Glutathione (GSH) and catalase/superoxide exhibited minor or no protective effect on binimetinib-induced enzyme inactivation. Trapping experiment by GSH induced a detection of GSH adduct, of which the formation was believed to be through the oxidation of electron-rich 1,4-benzenediamine to reactive 1,4-diiminoquinone species. Cytochrome P450 3A4, 2C9, and 2D6 were involved in the bioactivation of binimetinib. In conclusion, binimetinib was proven to display reversible and time-dependent inhibitory effect on CYP1A2, which may have implications for the toxicity of binimetinib.
Topics: Benzimidazoles; Cytochrome P-450 CYP1A2; Cytochrome P-450 Enzyme Inhibitors; Dose-Response Relationship, Drug; Humans; Microsomes, Liver; Molecular Structure; Time Factors
PubMed: 33728909
DOI: 10.1021/acs.chemrestox.1c00036 -
European Journal of Pharmaceutical... Jun 2021The cynomolgus monkey is a nonhuman primate that is often used for pharmacokinetic and toxicokinetic studies of new chemical entities. Species differences in drug...
The cynomolgus monkey is a nonhuman primate that is often used for pharmacokinetic and toxicokinetic studies of new chemical entities. Species differences in drug metabolism are obstacles for the extrapolation of animal data to humans. This study aimed to characterize hydrolase activities for typical compounds by cynomolgus monkey liver microsomes and recombinant monkey carboxylesterases (CES1 and CES2) and arylacetamide deacetylase (AADAC) compared with the activities in humans. To estimate the contribution of each hydrolase, the ratios of the expression level of each hydrolase in the liver microsomes and recombinant systems were used. For almost all of the tested human CES1 substrates, hydrolase activities in cynomolgus monkey liver microsomes tended to be lower than those in human liver microsomes, and recombinant cynomolgus monkey CES1 showed catalytic activity, but not for all substrates. For human CES2 substrates, hydrolase activities in cynomolgus monkey liver were higher than those in human liver microsomes, and recombinant monkey CES2 was responsible for their hydrolysis. Among human AADAC substrates, phenacetin was mainly hydrolyzed by monkey AADAC, whereas indiplon and ketoconazole were hydrolyzed by AADAC and other unknown enzymes. Flutamide was hydrolyzed by monkey CES2, not by AADAC. Rifamycins were hardly hydrolyzed in monkey liver microsomes. In conclusion, this study characterized the hydrolase activities of cynomolgus monkeys compared with those in humans. The findings would be helpful for pharmacokinetic or toxicokinetic studies of new chemical entities whose main metabolic pathway is hydrolysis.
Topics: Animals; Carboxylic Ester Hydrolases; Flutamide; Hydrolases; Hydrolysis; Liver; Macaca fascicularis; Microsomes, Liver
PubMed: 33722734
DOI: 10.1016/j.ejps.2021.105807 -
Molecular Pharmaceutics Mar 2021Hydrolytic reactions constitute an important pathway of drug metabolism and a significant route of prodrug activation. Many ophthalmic drugs and prodrugs contain ester...
Hydrolytic reactions constitute an important pathway of drug metabolism and a significant route of prodrug activation. Many ophthalmic drugs and prodrugs contain ester groups that greatly enhance their permeation across several hydrophobic barriers in the eye before the drugs are either metabolized or released, respectively, hydrolysis. Thus, the development of ophthalmic drug therapy requires the thorough profiling of substrate specificities, activities, and expression levels of ocular esterases. However, such information is scant in the literature, especially for preclinical species often used in ophthalmology such as rabbits and pigs. Therefore, our aim was to generate systematic information on the activity and expression of carboxylesterases (CESs) and arylacetamide deacetylase (AADAC) in seven ocular tissue homogenates from these two species. The hydrolytic activities were measured using a generic esterase substrate (4-nitrophenyl acetate) and, in the absence of validated substrates for rabbit and pig enzymes, with selective substrates established for human CES1, CES2, and AADAC (d-luciferin methyl ester, fluorescein diacetate, procaine, and phenacetin). Kinetics and inhibition studies were conducted using these substrates and, again due to a lack of validated rabbit and pig CES inhibitors, with known inhibitors for the human enzymes. Protein expression levels were measured using quantitative targeted proteomics. Rabbit ocular tissues showed significant variability in the expression of CES1 (higher in cornea, lower in conjunctiva) and CES2 (higher in conjunctiva, lower in cornea) and a poor correlation of CES expression with hydrolytic activities. In contrast, pig tissues appear to express only CES1, and CES3 and AADAC seem to be either low or absent, respectively, in both species. The current study revealed remarkable species and tissue differences in ocular hydrolytic enzymes that can be taken into account in the design of esterase-dependent prodrugs and drug conjugates, the evaluation of ocular effects of systemic drugs, and in translational and toxicity studies.
Topics: Animals; Carboxylesterase; Eye; Female; Humans; Hydrolysis; Male; Nitrophenols; Prodrugs; Proteomics; Rabbits; Substrate Specificity; Swine
PubMed: 33595329
DOI: 10.1021/acs.molpharmaceut.0c01154 -
Talanta Apr 2021MIL-101(Cr) and graphene aerogel (GA) were hybridized as a multifunctional adsorbent for the preconcentration of trace analytes. The novel MIL-101(Cr)@GA was prepared...
MIL-101(Cr) and graphene aerogel (GA) were hybridized as a multifunctional adsorbent for the preconcentration of trace analytes. The novel MIL-101(Cr)@GA was prepared and characterized by FTIR, XRD and SEM, where GA is serving as a carrier for MIL-101(Cr). The synthesized composite as a solid phase extraction (SPE) sorbent combined with UPLC-MS/MS was applied for the determination and quantification of five NSAIDs (phenacetin, meloxicam, naproxen, diclofenac sodium and carprofen) in different environmental water samples. The parameters influencing the whole extraction process were systematically optimized. Under the most favorable. conditions, good sensitivity was achieved with a limit of detection between 0.006 and 0.012 ng mL, the linear range of 0.02-2 ng mL for phenacetin, meloxicam and 0.05-5 ng mL for naproxen, diclofenac sodium, carprofen (r ≥ 0.9940). The satisfactory recoveries of the target analytes were in the range from 77.2 to 103.3% with relative standard deviation (RSD) from 0.6% to 8.4%. The established method was proved to be simple, highly sensitive and accurate. The adsorbent could be reusable up to nine cycles without any decrease in performance, which provides economic strategy and little waste generation. Adsorption behaviors were explored by choosing two typical types of NSAIDs as models to measure the adsorption capacity of MIL-101(Cr)@GA. The maximum adsorption capacities for PHE and NAP were 232.5 and 333.3 mg g, respectively.
Topics: Adsorption; Anti-Inflammatory Agents, Non-Steroidal; Chromatography, High Pressure Liquid; Chromatography, Liquid; Graphite; Metal-Organic Frameworks; Solid Phase Extraction; Tandem Mass Spectrometry; Water; Water Pollutants, Chemical
PubMed: 33592689
DOI: 10.1016/j.talanta.2020.121846 -
Alcohol (Fayetteville, N.Y.) Nov 2021Alcohol abuse has become a serious health issue worldwide. Ketamine can reduce addiction risk among patients with alcohol use disorders. This study aimed to determine...
BACKGROUND
Alcohol abuse has become a serious health issue worldwide. Ketamine can reduce addiction risk among patients with alcohol use disorders. This study aimed to determine the effects of alcohol on the pharmacokinetics of ketamine during long-term alcohol exposure.
METHOD
An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for determination of ketamine and norketamine was developed and validated. A total of 15 rats were given 40% alcohol for 3 weeks. The pharmacokinetics of ketamine were measured at time zero, 1 week, 2 weeks, and 3 weeks after alcohol exposure. The metabolic capability of liver CYP450 was evaluated using three probe drugs: metoprolol, phenacetin, and tolbutamide.
RESULTS
During drinking of 40% alcohol, the AUC(0-t), AUC(0-∞), and Cmax of ketamine and norketamine significantly increased, while V and CL significantly decreased with time (p < 0.001). The pharmacokinetic changes of norketamine were highly consistent with ketamine. Additionally, the concentration ratio of norketamine/ketamine in sample time also decreased over time. However, there were no pharmacokinetic changes of three probe drugs, which indicated there was no significant change of liver CYPs activities.
CONCLUSION
Alcohol significantly increases plasma concentration of ketamine and norketamine. The effect of alcohol on pharmacokinetics of ketamine should be considered in clinical therapy.
Topics: Alcoholism; Animals; Chromatography, Liquid; Humans; Ketamine; Rats; Tandem Mass Spectrometry
PubMed: 33549609
DOI: 10.1016/j.alcohol.2021.01.008 -
Drug Metabolism and Disposition: the... Apr 2021Human arylacetamide deacetylase (AADAC) plays a role in the detoxification or activation of drugs and is sometimes involved in the incidence of toxicity by catalyzing...
Human arylacetamide deacetylase (AADAC) plays a role in the detoxification or activation of drugs and is sometimes involved in the incidence of toxicity by catalyzing hydrolysis reactions. AADAC prefers compounds with relatively small acyl groups, such as acetyl groups. Eslicarbazepine acetate, an antiepileptic drug, is a prodrug rapidly hydrolyzed to eslicarbazepine. We sought to clarify whether AADAC might be responsible for the hydrolysis of eslicarbazepine acetate. Eslicarbazepine acetate was efficiently hydrolyzed by human intestinal and liver microsomes and recombinant human AADAC. The hydrolase activities in human intestinal and liver microsomes were inhibited by epigallocatechin gallate, a specific inhibitor of AADAC, by 82% and 88% of the control, respectively. The hydrolase activities in liver microsomes from 25 human livers were significantly correlated ( = 0.87, < 0.001) with AADAC protein levels, suggesting that the enzyme AADAC is responsible for the hydrolysis of eslicarbazepine acetate. The effects of genetic polymorphisms of on eslicarbazepine acetate hydrolysis were examined by using the constructed recombinant AADAC variants with T74A, V172I, R248S, V281I, N366K, or X400Q. AADAC variants with R248S or X400Q showed lower activity than wild type (5% or 21%, respectively), whereas those with V172I showed higher activity than wild type (174%). Similar tendencies were observed in the other four substrates of AADAC; that is, -nitrophenyl acetate, ketoconazole, phenacetin, and rifampicin. Collectively, we found that eslicarbazepine acetate is specifically and efficiently hydrolyzed by human AADAC, and several polymorphic alleles would be a factor affecting the enzyme activity and drug response. SIGNIFICANCE STATEMENT: This is the first study to clarify that arylacetamide deacetylase (AADAC) is responsible for the activation of eslicarbazepine acetate, an antiepileptic prodrug, to eslicarbazepine, an active form, in the human liver and intestines. In addition, we found that several polymorphic alleles would be a factor affecting the enzyme activity and drug response.
Topics: Adult; Aged; Carboxylic Ester Hydrolases; Cells, Cultured; Dibenzazepines; Enzyme Activation; Female; Humans; Hydrolases; Hydrolysis; Male; Microsomes, Liver; Middle Aged; Polymorphism, Genetic
PubMed: 33446525
DOI: 10.1124/dmd.120.000295 -
Current Drug Metabolism 2021Human cytochrome P450 enzyme 1A2 (CYP1A2) is one of the most important cytochrome P450 (CYP) enzymes in the liver, accounting for 13% to 15% of hepatic CYP enzymes.... (Review)
Review
Human cytochrome P450 enzyme 1A2 (CYP1A2) is one of the most important cytochrome P450 (CYP) enzymes in the liver, accounting for 13% to 15% of hepatic CYP enzymes. CYP1A2 metabolises many clinical drugs, such as phenacetin, caffeine, clozapine, tacrine, propranolol, and mexiletine. CYP1A2 also metabolises certain precarcinogens such as aflatoxins, mycotoxins, nitrosamines, and endogenous substances such as steroids. The regulation of CYP1A2 is influenced by many factors. The transcription of CYP1A2 involves not only the aromatic hydrocarbon receptor pathway but also many additional transcription factors, and CYP1A2 expression may be affected by transcription coactivators and compression factors. Degradation of CYP1A2 mRNA and protein, alternative splicing, RNA stability, regulatory microRNAs, and DNA methylation are also known to affect the regulation of CYP1A2. Many factors can lead to changes in the activity of CYP1A2. Smoking, polycyclic aromatic hydrocarbon ingestion, and certain drugs (e.g., omeprazole) increase its activity, while many clinical drugs such as theophylline, fluvoxamine, quinolone antibiotics, verapamil, cimetidine, and oral contraceptives can inhibit CYP1A2 activity. Here, we review the drugs metabolised by CYP1A2, the metabolic mechanism of CYP1A2, and various factors that influence CYP1A2 metabolism. The metabolic mechanism of CYP1A2 is of great significance in the development of personalised medicine and CYP1A2 target-based drugs.
Topics: Animals; Cytochrome P-450 CYP1A2; Cytochrome P-450 CYP1A2 Inhibitors; Enzyme Inhibitors; Humans
PubMed: 33397254
DOI: 10.2174/1389200221999210101233135 -
Chemosphere Apr 2021Acetaminophen (ACT) and phenacetin (PNT) removal during light-emitting diode (LED)-UV photolysis of persulfate (PS) was evaluated with a typical wavelength of 365 nm....
Acetaminophen (ACT) and phenacetin (PNT) removal during light-emitting diode (LED)-UV photolysis of persulfate (PS) was evaluated with a typical wavelength of 365 nm. Decay of PNT and ACT in pH ranges of 5.5-8.5 followed pseudo-first order kinetics. Maximum pseudo-first order rate constants (k) of ACT and PNT decomposition of 1.8 × 10 and 1.2 × 10 min, respectively, were obtained at pH 8.5. Hydroxyl radicals (·OH), sulfate radicals (SO·), superoxide radicals (O·), and singlet oxygen (O) were determined in-situ electron paramagnetic resonance (EPR) and alcohol scavenging tests. The average contributions of ·OH and SO· were 23.5% and 53.0% for PNT removal, and 15.9% and 53.0% for ACT removal at pH ranges of 5.5-8.5. In samples subjected to chlorination after LED-UV/PS pre-oxidation, a relatively small total concentration of five halogenated disinfection by-products (DBPs) was obtained of 90.9 μg L (pH 5.5) and 126.7 μg L (pH 7.0), which is 58.5% and 30.2% lower than that in system without LED-UV/PS pre-oxidation. Meanwhile, a higher maximum value of total DBP concentration was obtained at pH 8.5 (445.6 μg L) following LED-UV/PS pre-oxidation. The results of economy evaluation showed that UV was more cost-effective in application for organic contaminant removal compared with UV.
Topics: Acetaminophen; Kinetics; Oxidation-Reduction; Phenacetin; Photolysis; Reactive Oxygen Species; Ultraviolet Rays; Water Pollutants, Chemical; Water Purification
PubMed: 33387793
DOI: 10.1016/j.chemosphere.2020.129337 -
Biopharmaceutics & Drug Disposition Jan 2021CYP1A2 is one of the main Cytochrome P450 enzymes in the human liver associated with the metabolism of several xenobiotics. CYP1A2 is especially involved in the...
CYP1A2 is one of the main Cytochrome P450 enzymes in the human liver associated with the metabolism of several xenobiotics. CYP1A2 is especially involved in the metabolic activation of different procarcinogens. Therefore, the development of cancer may be inhibited by inhibiting CYP1A2 activity. Here, the inhibitory effect of HYIpro-3-1 and its derivatives on CYP1A2 activity in human liver microsomes (HLM) was studied through LC-MS/MS using a cocktail assay. Among the four compounds, HYIpro-3-1 showed the most selective and strongest inhibitory effect on CYP1A2 at IC values of 0.1 µM in HLMs and inhibition was confirmed using purified human CYP1A2. It was determined that inhibition is reversible because the inhibitory effect of HYIpro-3-1 is not dependent on preincubation time. HYIpro-3-1 showed a typical pattern of competitive inhibition for CYP1A2-catalyzed phenacetin O-deethylation, based on the Lineweaver-Burk plot, with a Ki value of 0.05 μM in HLMs; the secondary plot also showed a linear pattern. In our study, HYIpro-3-1 was proposed as a novel inhibitor with the capacity to selectively inhibit CYP1A activity in HLMs.
Topics: Cytochrome P-450 CYP1A1; Cytochrome P-450 CYP1A2 Inhibitors; Humans; Microsomes, Liver
PubMed: 33386627
DOI: 10.1002/bdd.2259 -
Chemosphere Jan 2021Cobalt doped iron oxychloride (Co-FeOCl) was synthesized and employed as catalyst in Fenton degradation of paracetamol (APAP) and phenacetin (PNCT) for the first time....
Cobalt doped iron oxychloride (Co-FeOCl) was synthesized and employed as catalyst in Fenton degradation of paracetamol (APAP) and phenacetin (PNCT) for the first time. The catalytic performance was evaluated by means of various parameters including catalyst load, hydrogen peroxide (HO) dose and pH value. The high removal of APAP (87.5%) and PNCT (76.0%) was obtained under conditions of 0.2 g/L Co-FeOCl and 0.5 mM HO at pH 7.0, with calculated pseudo-first order kinetic constants of 0.031 min for APAP and 0.023 min for PNCT. Particularly, quenching tests and in situ electron spin resonance (ESR) tests were employed for the identification of the reactive oxygen species (ROS) in system. Hydroxyl radical (·OH) and superoxide radical (O·) were the primary ROS in Co-FeOCl/HO system. A possible mechanism for HO activation by Co-FeOCl catalyst was proposed as well. Finally, the formation of typical disinfection by-products (DBPs) decreased slightly in Co-FeOCl/HO pre-oxidation. However, stability and reusability of Co-FeOCl were deactivated in the consecutive three cycles.
Topics: Acetaminophen; Catalysis; Cobalt; Hydrogen Peroxide; Iron Compounds; Oxidation-Reduction; Phenacetin
PubMed: 33297032
DOI: 10.1016/j.chemosphere.2020.127989