Comparison of In Vitro Hepatic Scoparone 7-O-Demethylation between Humans and Experimental Animals

Planta Medica ◽  
2017 ◽  
Vol 84 (05) ◽  
pp. 320-328 ◽  
Author(s):  
Anam Fayyaz ◽  
Seddy Makwinja ◽  
Seppo Auriola ◽  
Hannu Raunio ◽  
Risto Juvonen
Keyword(s):  
G Protein ◽  
Liver Microsomes ◽  
Herbal Medicines ◽  
Bioactive Compound ◽  
Assay Method ◽  
Primary Metabolism ◽  
Rat Liver Microsomes ◽  
Cyp Enzymes ◽  
Rate Of Reaction

AbstractScoparone is a natural bioactive compound in Chinese herbal medicines. It has numerous pharmacological actions, including liver protective, hypolipidemic, antitumor, and anti-inflammatory effects. The primary metabolism route of scoparone is O-demethylation to scopoletin or isoscopoletin catalyzed by CYP enzymes. The aims of our study were to identify the human CYP enzymes catalyzing scoparone 7-O-demethylation to scopoletin and to compare this oxidation reaction in liver microsomes among different species. A high throughput fluorescent-based assay method was developed to determine the scoparone 7-O-demethylation to scopoletin rate. The rate was 100 – 400 nmol/(min×g protein) in mouse and rabbit liver microsomes, 10 – 20 nmol/(min×g protein) in pig microsomes, 1 – 3 nmol/(min×g protein) in human and less than 1 nmol/(min×g protein) in rat liver microsomes. Human CYP1A1 (Km 13 µM and Vmax 0.8 min−1), CYP1A2 (Km 48 µM and Vmax 0.3 min−1), and CYP2A13 (Km 10 µM and Vmax 22 min−1) were the most efficient catalysts of the reaction. The CYP2A6 selective inhibitor pilocarpine and an antibody against mouse CYP2A5 inhibited scoparone 7-O-demethylation to scopoletin in rabbit, mouse, and pig liver microsomes, indicating involvement of CYP2A enzymes in the reaction. Hepatic scoparone 7-O-demethylation to scopoletin differed between species both with respect to the rate of reaction and catalyzing enzymes. These species differences need to be taken into account when testing scoparone pharmacokinetics in animals and humans.

scholarly journals The Effects of Japanese Herbal Medicines on Drug Oxidations in Rat Liver Microsomes in vitro.

2002 ◽  
Vol 28 (4) ◽  
pp. 360-365 ◽  
Author(s):  
Maharu Kimura ◽  
Atsushi Hasegawa ◽  
Hiroyoshi Nakamura ◽  
Shigeru Ohmori ◽  
Itsuko Ishii ◽  
...  
Keyword(s):  
Rat Liver ◽  
Liver Microsomes ◽  
Herbal Medicines ◽  
Rat Liver Microsomes

scholarly journals Inhibition of UGT2B7 Enzyme Activity in Human and Rat Liver Microsomes by Herbal Constituents

Molecules ◽  
2018 ◽  
Vol 23 (10) ◽  
pp. 2696 ◽  
Author(s):  
Nurul Abdullah ◽  
Sabariah Ismail
Keyword(s):  
Rat Liver ◽  
Liver Microsomes ◽  
Herbal Medicines ◽  
Inhibitory Effect ◽  
Intrinsic Clearance ◽  
Rat Liver Microsomes ◽  
Metabolizing Enzymes ◽  
Conventional Drug ◽  
Ic50 Values

The co-use of conventional drug and herbal medicines may lead to herb-drug interaction via modulation of drug-metabolizing enzymes (DMEs) by herbal constituents. UDP-glucuronosyltransferases (UGTs) catalyzing glucuronidation are the major metabolic enzymes of Phase II DMEs. The in vitro inhibitory effect of several herbal constituents on one of the most important UGT isoforms, UGT2B7, in human liver microsomes (HLM) and rat liver microsomes (RLM) was investigated. Zidovudine (ZDV) was used as the probe substrate to determine UGT2B7 activity. The intrinsic clearance (Vmax/Km) of ZDV in HLM is 1.65 µL/mg/min which is ten times greater than in RLM, which is 0.16 µL/mg/min. Andrographolide, kaempferol-3-rutinoside, mitragynine and zerumbone inhibited ZDV glucuronidation in HLM with IC50 values of 6.18 ± 1.27, 18.56 ± 8.62, 8.11 ± 4.48 and 4.57 ± 0.23 µM, respectively, hence, herb-drug interactions are possible if andrographolide, kaempferol-3-rutinoside, mitragynine and zerumbone are taken together with drugs that are highly metabolized by UGT2B7. Meanwhile, only mitragynine and zerumbone inhibited ZDV glucuronidation in RLM with IC50 values of 51.20 ± 5.95 μM and 8.14 ± 2.12 µM, respectively, indicating a difference between the human and rat microsomal model so caution must be exercised when extrapolating inhibitory metabolic data from rats to humans.

scholarly journals Nontargeted Metabolomics by High-Resolution Mass Spectrometry to Study the In Vitro Metabolism of a Dual Inverse Agonist of Estrogen-Related Receptors β and γ, DN203368

Pharmaceutics ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 776
Author(s):  
Sin-Eun Kim ◽  
Seung-Bae Ji ◽  
Euihyeon Kim ◽  
Minseon Jeong ◽  
Jina Kim ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Resolution ◽  
Human Liver ◽  
High Resolution Mass Spectrometry ◽  
Liver Microsomes ◽  
Human Liver Microsomes ◽  
Rat Liver Microsomes ◽  
High Resolution Mass ◽  
Resolution Mass

DN203368 ((E)-3-[1-(4-[4-isopropylpiperazine-1-yl]phenyl) 3-methyl-2-phenylbut-1-en-1-yl] phenol) is a 4-hydroxy tamoxifen analog that is a dual inverse agonist of estrogen-related receptor β/γ (ERRβ/γ). ERRγ is an orphan nuclear receptor that plays an important role in development and homeostasis and holds potential as a novel therapeutic target in metabolic diseases such as diabetes mellitus, obesity, and cancer. ERRβ is also one of the orphan nuclear receptors critical for many biological processes, such as development. We investigated the in vitro metabolism of DN203368 by conventional and metabolomic approaches using high-resolution mass spectrometry. The compound (100 μM) was incubated with rat and human liver microsomes in the presence of NADPH. In the metabolomic approach, the m/z value and retention time information obtained from the sample and heat-inactivated control group were statistically evaluated using principal component analysis and orthogonal partial least-squares discriminant analysis. Significant features responsible for group separation were then identified using tandem mass spectra. Seven metabolites of DN203368 were identified in rat liver microsomes and the metabolic pathways include hydroxylation (M1-3), N-oxidation (M4), N-deisopropylation (M5), N,N-dealkylation (M6), and oxidation and dehydrogenation (M7). Only five metabolites (M2, M3, and M5-M7) were detected in human liver microsomes. In the conventional approach using extracted ion monitoring for values of mass increase or decrease by known metabolic reactions, only five metabolites (M1-M5) were found in rat liver microsomes, whereas three metabolites (M2, M3, and M5) were found in human liver microsomes. This study revealed that nontargeted metabolomics combined with high-resolution mass spectrometry and multivariate analysis could be a more efficient tool for drug metabolite identification than the conventional approach. These results might also be useful for understanding the pharmacokinetics and metabolism of DN203368 in animals and humans.

In vitro metabolism in Sprague–Dawley rat liver microsomes of forsythoside A in different compositions of Shuang–Huang–Lian

Fitoterapia ◽  
2011 ◽  
Vol 82 (8) ◽  
pp. 1222-1230 ◽  
Author(s):  
Wei Zhou ◽  
Liu-qing Di ◽  
Jin-jun Shan ◽  
Xiao-lin Bi ◽  
Le-tian Chen ◽  
...  
Keyword(s):  
Rat Liver ◽  
Liver Microsomes ◽  
In Vitro Metabolism ◽  
Rat Liver Microsomes ◽  
Sprague Dawley ◽  
Sprague Dawley Rat

scholarly journals In Vitro Characterization of Borneol Metabolites by GC--MS Upon Incubation with Rat Liver Microsomes

2008 ◽  
Vol 46 (5) ◽  
pp. 419-423 ◽  
Author(s):  
R. Zhang ◽  
C.-h. Liu ◽  
T.-l. Huang ◽  
N.-s. Wang ◽  
S.-q. Mi
Keyword(s):  
Rat Liver ◽  
Liver Microsomes ◽  
Rat Liver Microsomes ◽  
In Vitro Characterization

In Vitro Metabolism of E2, G2: Novel Bile Acid-Coupling Camptothecin Analogues, in Rat Liver Microsomes

2021 ◽  
Vol 16 ◽  
Author(s):  
Xiangli Zhang ◽  
Qin Shen ◽  
Yi Wang ◽  
Leilei Zhou ◽  
Qi Weng ◽  
...  
Keyword(s):  
Bile Acid ◽  
Phase I ◽  
Rat Liver ◽  
Deoxycholic Acid ◽  
Liver Microsomes ◽  
Intrinsic Clearance ◽  
Rat Liver Microsomes ◽  
Camptothecin Analogues

Background: E2 (Camptothecin - 20 (S) - O- glycine - deoxycholic acid), and G2 (Camptothecin - 20 (S) - O - acetate - deoxycholic acid) are two novel bile acid-derived camptothecin analogues by introducing deoxycholic acid in 20-position of CPT(camptothecin) with greater anticancer activity and lower systematic toxicity in vivo. Objective: We aimed to investigate the metabolism of E2 and G2 by Rat Liver Microsomes (RLM). Methods: Phase Ⅰ and Phase Ⅱ metabolism of E2 and G2 in rat liver microsomes were performed respectively, and the mixed incubation of phase I and phase Ⅱ metabolism of E2 and G2 was also processed. Metabolites were identified by liquid chromatographic/mass spectrometry. Results: The results showed that phase I metabolism was the major biotransformation route for both E2 and G2. The isoenzyme involved in their metabolism had some difference. The intrinsic clearance of G2 was 174.7mL/min. mg protein, more than three times of that of E2 (51.3 mL/min . mg protein), indicating a greater metabolism stability of E2. 10 metabolites of E2 and 14 metabolites of G2 were detected, including phase I metabolites (mainly via hydroxylations and hydrolysis) and their further glucuronidation products. Conclusion: These findings suggested that E2 and G2 have similar biotransformation pathways except some difference in the hydrolysis ability of the ester bond and amino bond from the parent compounds, which may result in the diversity of their metabolism stability and responsible CYPs(Cytochrome P450 proteins).

scholarly journals Determination of in Vivo Biological Activities of Dodonaea Viscosa Flowers Against CCL4 Toxicity in Albino Mice With Bioactive Compound Detection

2021 ◽  
Author(s):  
Hina Gul ◽  
Muhammad Awais ◽  
Salina Saddick ◽  
Falak Sher Khan ◽  
Muhammad Gulfraz ◽  
...  
Keyword(s):  
Protective Effect ◽  
Biological Activities ◽  
Hematological Parameters ◽  
Octadecenoic Acid ◽  
Radical Scavenging ◽  
Herbal Medicines ◽  
Bioactive Compound ◽  
Dodonaea Viscosa

Abstract Dodonaea viscosa L. Jacq. is an evergreen shrub and native to Asia, Africa and Australia. It has been used as traditional medicine in different countries. The foremost objective of the current study was to discover protective potential of D. Viscosa flowers Methanol (DVM) and Chloroform (DVC) extracts against CCL4 induced toxicity in mice. This study was intended to identify phytochemicals through HPLC, GCMS and FT-IR as well as in vitro antioxidant and in vitro antituberculosis activity. Our comprehensive findings indicate that Dodonaea viscosa is valuable and widespread herbal medicines through therapeutic potentials for curing various ailments. Dodonaea viscosa flowers are found to have protective effect against oxidative stress produced by CCL4 in liver, kidney and spleen. The level of hepatic enzymes (ALP, AST ALT and Direct bilirubin), hematological parameters (RBCs, WBCs and Platelets), total protein and liver antioxidant enzymes (SOD, GPx and CAT) were restored by the intake of DV extracts after decline in levels by CCL4. Histopathological results discovered the defensive effect of 300mg/kg of DVM extract against CCL4 induced damage, thus having improved protective effect as compared to DVC and control. As a result of analysis total flavonoids and total phenolics were also revealed. Phytochemical investigation by HPLC identified gallic acid, epicatechin, cumeric acid, flavonoids while Oleic acid (Octadecenoic acid) (C18H34O2), Stearic acid (C18H36O2), Ricinoleic acid (C18H34O3) and Cedrol (C15H26O) was estimated by GCMS. DVM extract exhibited resistance against in vitro Mycobacterium tuberculosis strains. This study proposed that protective effect of DV against oxidative damage induced in Liver, Kidney and Spleen can possibly be correlated to their antioxidant as well as free radical scavenging property.

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