The Development and Validation of a Novel “Dual Cocktail” Probe for Cytochrome P450s and Transporter Functions to Evaluate Pharmacokinetic Drug-Drug and Herb-Drug Interactions

Abstract

This study was designed to develop and validate a 10 probe drug cocktail named “Dual Cocktail”, composed of caffeine (Cyp1a2 in rat and CYP1A2 in human, 1 mg/kg), diclofenac (Cyp2c11 in rat and CYP2C9 in human, 2 mg/kg), omeprazole (Cyp2c11 in rat and CYP2C19 in human, 2 mg/kg), dextromethorphan (Cyp2d2 in rat and CYP2D6 in human, 10 mg/kg), nifedipine (Cyp3a1 in rat and CYP3A4 in human, 0.5 mg/kg), metformin (Oct1/2 in rat and OCT1/2 in human, 0.5 mg/kg), furosemide (Oat1/3 in rat and OAT1/3 in human, 0.1 mg/kg), valsartan (Oatp2 in rat and OATP1B1/1B3 in human, 0.2 mg/kg), digoxin (P-gp in rat and human, 2 mg/kg), and methotrexate (Mrp2 in rat and MRP2 in human, 0.5 mg/kg), for the evaluation of pharmacokinetic drug–drug and herb-drug interactions through the modulation of a representative panel of CYP enzymes or transporters in rats. To ensure no interaction among the ten probe substrates, we developed a 2-step evaluation protocol. In the first step, the pharmacokinetic properties of five individual CYP probe substrates and five individual transporter substrates were compared with the pharmacokinetics of five CYP cocktail or five transporters cocktails in two groups of randomly assigned rats. Next, a pharmacokinetic comparison was conducted between the CYP or transporter cocktail group and the dual cocktail group, respectively. None of the ten comparison groups was found to be statistically significant, indicating the CYP and transporter substrate sets or dual cocktail set could be concomitantly administered in rats. The “Dual Cocktail” was further validated by assessing the metabolism of nifedipine and omeprazole, which was significantly reduced by a single oral dose of ketoconazole (10 mg/kg); however, no changes were observed in the pharmacokinetic parameters of other probe substrates. Additionally, multiple oral doses of rifampin (20 mg/kg) reduced the plasma concentrations of nifedipine and digoxin, although not any of the other substrates. In conclusion, the dual cocktail can be used to characterize potential pharmacokinetic drug–drug interactions by simultaneously monitoring the activity of multiple CYP isoforms and transporters.

Involvement of Organic Anion Transporters in the Pharmacokinetics and Drug Interaction of Rosmarinic Acid

“…We evaluated the inhibitory effect of rosmarinic acid on OCT1, OAT3, OATP1B1, and OATP1B3 by investigating the pharmacokinetic interaction between rosmarinic acid and the probe substrates for these transporters. Furosemide was selected as a probe substrate for OAT1 and OAT3 [ 20 , 25 ] and valsartan was selected as a probe substrate for OATP1B1 and OATP1B3 [ 20 ]. The plasma concentrations of furosemide acid were increased by the co-administration of rosmarinic acid ( Figure 6 A).…”

“…Valsartan concentrations were analyzed using an Agilent 6430 Triple Quadrupole LC-MS/MS system utilizing a modification of a previously published method [ 20 , 23 ]. The separation was performed on a Polar RP column (2.0 × 150 mm, 5 μm) using a mobile phase consisting of water and methanol (25:75 v / v ) with 0.1% formic acid at a flow rate of 0.2 mL/min.…”

“…The results suggested the involvement of drug transporters that are expressed mainly in the kidney in the pharmacokinetics and tissue distribution. Drug transporters, alone or in combination with drug-metabolizing enzymes, have drawn much attention in herb-drug interactions because they regulate the absorption, distribution, and elimination of drugs [ 19 , 20 ]. Therefore, the objectives of this study were to investigate the involvement of drug transporters in the pharmacokinetics of rosmarinic acid and to investigate the transporter-mediated drug interaction potential of rosmarinic acid.…”

“…Therefore, the objectives of this study were to investigate the involvement of drug transporters in the pharmacokinetics of rosmarinic acid and to investigate the transporter-mediated drug interaction potential of rosmarinic acid. In this study, we included solute carrier transporters such as organic anion transporter (OAT)1, OAT3, organic anion transporting polypeptide (OATP)1B1, OATP1B3, organic cation transporter (OCT)1, OCT2, and Na + /taurocholate cotransporting polypeptide (NTCP) that are clinically important in terms of drug–drug interactions and herb–drug interaction [ 20 , 21 ].…”

Evaluation of Efficacy and Safety After Replacement of Methyl Hydrogen With Deuterium at 7-position Methyl Formate of Clopidogrel

“…Sedimented liver microsomes were resuspended in cold potassium pyrophosphate solution(PH7.4), and centrifuged at 100,000 g 4℃for 60min, sedimented liver microsomes were resuspended in cold Tris-HCL(PH7.4), stored separately at -80℃ for detection. The concentration of liver microsomes was analyzed by BCA method, the activity of cytochrome P450(CYP450) enzymes was determined by Cocktail method [24][25] , phenacetin, bupropion, paclitaxel, tolbutamide, S-mephenytoin, dextromethorphan and testosterone were used as probe substrates for CYP1A2(mice:CYP1a2), CYP2B6(mice:CYP2b9), CYP2C8(CYP2c29), CYP2C9(mice:CYP2c37), CYP2C19(mice:CYP2c39/40/44), CYP2D6(mice:CYP2d6) and CYP3A4(mice:CYP3a11) respectively.…”