Differential effect of Shenmai injection, a herbal preparation, on the cytochrome P450 3A-mediated 1′-hydroxylation and 4-hydroxylation of midazolam

Xia, Chunhua; Sun, Jianguo; Shang, Lili; Zhang, Xiaoxuan; Zhang, Rong; Wang, Xiaojin; Hao, Haiping; Xie, Lin

doi:10.1016/j.cbi.2009.03.022

Cited by 21 publications

(20 citation statements)

References 27 publications

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“…of three determinations performed in duplicate. dmd.aspetjournals.org modulators could exert different effects on each metabolite formation (Xia et al, 2009;Dong et al, 2011). As reported, erlotinib and panaxytriol, an active component in Shenmai injection (Zeng et al, 2013), both showed significant activation on MDZ 19-hydroxylation, whereas the activation but not inhibition made it difficult for the in vivo extrapolation.…”

Section: Discussionmentioning

confidence: 84%

Deoxyschizandrin, a Naturally Occurring Lignan, Is a Specific Probe Substrate of Human Cytochrome P450 3A

Cao

Zhang

et al. 2013

Drug Metab Dispos

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To accurately predict the modifications done during metabolic processes by cytochrome P450 (P450) 3A enzyme, selecting substrates that best represent a broad range of substrate substitutions and that follow the Michaelis-Menten kinetic properties is highly necessary. In the present study, the oxidative pathways of deoxyschizandrin (DS), the most abundant lignan in Fructus Schisandrae fruit extract, were characterized with liver microsomes from human (HLM) and rat (RLM). Only one monohydroxylated metabolite 7(S)-hydroxylated metabolite (isoschizandrin, ISZ), was identified using liquid chromatography-mass spectrometry and nuclear magnetic resonance techniques. CYP3A4 and CYP3A5 were found to be the major isoforms involved in the monohydroxylation of DS. Also, the kinetic studies showed that DS hydroxylation obeyed MichaelisMenten kinetics both in HLM and in RLM. However, the subsequent metabolism of ISZ was nearly nonexistent when DS was present. More importantly, the interactions between DS and three well characterized CYP3A probe substrates, testosterone (TST), midazolam (MDZ), and nifedipine (NIF), were studied. TST and MDZ were shown to compete with DS for the mutual binding site, causing K m to be increased. The presence of DS also lowered the binding affinities for MDZ and TST. However, DS showed only slight inhibitory effects on nifedipine (NIF) oxidation even though NIF was able to inhibit DS hydroxylation in a noncompetitive fashion. These results show that DS is a good representative substrate of MDZ and TST primarily due to their shared, large binding regions on CYP3A. Therefore, DS is an attractive candidate as a novel CYP3A probe substrate for predicting the metabolic modifications in CYP3A activity.

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Section: Discussionmentioning

confidence: 84%

Deoxyschizandrin, a Naturally Occurring Lignan, Is a Specific Probe Substrate of Human Cytochrome P450 3A

Cao

Zhang

et al. 2013

Drug Metab Dispos

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“…In our previous study, panaxytriol showed differential effect on the metabolic pathways of MDZ, significantly inscreasing MDZ 1′-hydroxylation but inhibiting MDZ 4-hydroxylation. 11,12) This results revealed that PXT had the potential to inhibit or activate the CYP3A4 enzyme activity. Therefore, it is necessary to further study the impact mechanism of PXT on CYP3A4 enzyme and possible interaction with other clinical drugs.…”

mentioning

confidence: 75%

Mechanism of Action of Panaxytriol on Midazolam 1′-Hydroxylation and 4-Hydroxylation Mediated by CYP3A in Liver Microsomes and Rat Primary Hepatocytes

Zhang

Zeng

et al. 2015

Biological & Pharmaceutical Bulletin

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In our previous study, panaxytriol (PXT) was shown to enhance midazolam (MDZ) 1′-hydroxylation significantly but to inhibit MDZ 4-hydroxylation. To explore the underlying mechanism, we investigated the effects of PXT on cytochrome P450 3A (CYP3A)-mediated MDZ metabolic pathways using rat liver microsomes (RLM), human liver microsomes (HLM), and rat primary hepatocytes. In the presence of PXT, the V max of 4-OH MDZ decreased from 0.72 to 0.51 nmol/min·mg pro in RLM and from 0.32 to 0.12 nmol/ min·mg pro in HLM, and the K m value increased from 5.12 to 7.26 µM in RLM and from 27.87 to 32.80 µM in HLM. But the presence of PXT reduced the K m and increased the V max values of MDZ 1′-hydroxylation in RLM and HLM. Interestingly, the differential effect of PXT on MDZ 4-hydroxylation and 1′-hydroxylation was also observed in primary rat hepatocytes after 45-min culture. PXT did not affect the expression levels of CYP3A1/2 mRNA in rat hepatocytes. With extension of the culture time to 6 h, however, PXT significantly inhibited both MDZ 4-hydroxylation and 1′-hydroxylation, and the expression level of CYP3A1/2 mRNA was decreased to 87% and 80% (CYP3A1) and to 89% and 85% (CYP3A2) of those in controls in the presence of PXT 4.0 and 8.0 µg/mL, respectively. These results suggest that PXT could activate MDZ 1′-hydroxylation but inhibit MDZ 4-hydroxylation by changing the CYP3A enzyme affinity and metabolic rate after a shortterm intervention. However, long-term treatment with PXT could inhibit both the 4-hydroxylation and 1′-hydroxylation of MDZ by downregulating CYP3A1/2 mRNA expression.Key words panaxytriol; midazolam hydroxylation metabolism; cytochrome P450 3A; differential effect mechanism Cytochrome P450 (CYP450) is a superfamily of hemeproteins that involved in the biotransformation of the majority of clinical drugs. The inhibition or induction effect of clinical drugs on CYP450 is considered as one of the important factors leading to potential drug-drug interactions.1-3) CYP3A4 is one of the primary CYP450 enzymes in human liver and catalyzes the metabolism of more than 50% of clinical drugs including terfenadine, theophylline and astemizole, etc. Midazolam (MDZ) is the specific substrate of CYP3A4 enzyme and is mainly metabolized to 4-hydroxymidazolam (4-OH MDZ) and 1′-hydroxymidazolam (1′-OH MDZ) by combining with the two different sites of CYP3A4 enzyme active center.4,5) Therefore, the formation of the two metabolites are often simultaneously quantified for the assessment of CYP3A4 activity.Traditional Chinese medicine is widely used in clinical practice in China because of the advantages of good curative effect and less adverse reaction. The incidence of Chinese medicine and chemical medicine interactions significantly increased as the widely using of Chinese medicine, and the change of CYP450 enzyme activity is the principal cause resulting in drug metabolic interactions. 6,7) As a well known study, salvia extract exhibited a marked inhibition effect on the expression of CYP3A4 and CYP1A2 mRNA in HepG2, and high...

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“…SMI was provided by Hebei Shenwei Pharmaceutical Co. Ltd (Hebei, China). Each milliliter volume of SMI was comprised of the extract of 0.1 g red ginseng and 0.1 g radix ophiopogonis and contained 76.2 mg ginsenoside Rg1, 69.7 mg ginsenoside Re, 68.1 mg ginsenoside Rd, 178.4 mg ginsenoside Rb1, 15.7 mg ophiopogonin D, and 1.7 mg ophiopogonone A (Xia et al, 2009). Glucose-6-phosphate, NADP + , glucose-6-phosphate dehydrogenase, MDZ, 19-OH-MDZ and 4-OH-MDZ were purchased from Sigma-Aldrich (St. Louis, MO).…”

Section: Methodsmentioning

confidence: 99%

“…It is used to treat atherosclerotic coronary heart disease and viral myocarditis and is often concomitantly administered with other prescribed medications, including certain cardiovascular drugs with a narrow therapeutic window (Janetzky and Morreale, 1997;Gurley et al, 2002;Donovan et al, 2003;Jiang et al, 2004;Yuan et al, 2004). In our previous study (Xia et al, 2009), we demonstrated that SMI could significantly enhance 19-hydroxylation of MDZ but inhibit 4-hydroxylation of this compound in human liver microsomes (HLM), rat liver microsomes (RLM), and recombinant human CYP3A4. Furthermore, total ginsenoside, Ophiopogon total saponins, Ophiopogon total flavones, ginsenoside Rd, and ophiopogonone A (important constituents of SMI) were all found to significantly inhibit both 19-hydroxylation and 4-hydroxylation of MDZ in HLM and RLM.…”

Section: Introductionmentioning

confidence: 99%

“…Drug interactions involving substrates, inhibitors, activators, or inducers of CYP3A4 are more prevalent and complex than those of other members of the P450 family (Kim et al, 2006;Pal and Mitra, 2006). It is becoming evident that many CYP3A4-mediated reactions proceed with atypical MichaelisMenten kinetics, including substrate activation (Shou et al, 1999), substrate inhibition (Lin et al, 2001;Wu, 2011), and differential kinetics Zhang and Lim, 2008;Xia et al, 2009). These phenomena are thought to be due to the binding of multiple substrates within the active site of the enzyme (Kenworthy et al, 2001;Galetin et al, 2003;Zhou et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

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Identification of the Active Components in Shenmai Injection that Differentially Affect Cyp3a4-Mediated 1′-Hydroxylation and 4-Hydroxylation of Midazolam

Zeng

Xia

et al. 2013

Drug Metab Dispos

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Shenmai injection (SMI) is a popular herbal preparation that is widely used for the treatment of atherosclerotic coronary heart disease and viral myocarditis. In our previous study, SMI was shown to differentially affect CYP3A4-mediated 19-hydroxylation and 4-hydroxylation of midazolam (MDZ). The present study was conducted to identify the active components in SMI responsible for the differential effects on MDZ metabolism, using in vitro incubation systems (rat and human liver microsomes and a recombinant CYP3A4 system) to measure 19-hydroxylation and 4-hydroxylation of MDZ. First, different fractions of SMI were obtained by gradient elution on an solid phase extraction system and individually tested for their effects on MDZ metabolism. The results demonstrated that lipid-soluble constituents were likely to be the predominant active components of SMI. Second, the possible active components were gradually separated on an high-performance liquid chromatography system under different conditions and individually tested in vitro for their effects on MDZ metabolism. Third, the active component obtained in the above experiment was collected and subjected to structural analysis, and identified as panaxytriol (PXT). Finally, it was validated that PXT had significant differential effects on 19-hydroxylation and 4-hydroxylation of MDZ in various in vitro systems that were similar to those of SMI. We conclude that PXT is the constituent of SMI responsible for the differential effects on CYP3A4-mediated 19-hydroxylation and 4-hydroxylation of MDZ.

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Differential effect of Shenmai injection, a herbal preparation, on the cytochrome P450 3A-mediated 1′-hydroxylation and 4-hydroxylation of midazolam

Cited by 21 publications

References 27 publications

Deoxyschizandrin, a Naturally Occurring Lignan, Is a Specific Probe Substrate of Human Cytochrome P450 3A

Deoxyschizandrin, a Naturally Occurring Lignan, Is a Specific Probe Substrate of Human Cytochrome P450 3A

Mechanism of Action of Panaxytriol on Midazolam 1′-Hydroxylation and 4-Hydroxylation Mediated by CYP3A in Liver Microsomes and Rat Primary Hepatocytes

Identification of the Active Components in Shenmai Injection that Differentially Affect Cyp3a4-Mediated 1′-Hydroxylation and 4-Hydroxylation of Midazolam

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