Research ArticlesN-(3,4-dimethoxyphenethyl)-4-(6,7-dimethoxy-3,4-dihydroisoquinolin-2[1H]-yl)-6,7-dimethoxyquinazolin-2-amine (CP-100,356) as a “chemical knock-out equivalent” to assess the impact of efflux transporters on oral drug absorption in the rat
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INTRODUCTION
During initial stages of hit-to-lead phase in drug discovery, most new chemical entities (NCEs) suffer from poor systemic exposure after oral (p.o.) dosing in animals, necessitating a need for identifying issues limiting oral absorption. Besides first pass metabolism, other factors such aqueous solubility, membrane permeability, and active drug efflux can attenuate oral absorption. With regards to active drug efflux, members of the ATP-binding cassette (ABC) superfamily of efflux transporters
Materials
CP-100,356 (chemical and isomeric purity >99%) was synthesized at Pfizer Global Research and Development (Groton, CT). Detailed synthetic procedure for CP-100,356 and related analogs has been published.31 Solvents used for analysis were of analytical or HPLC grade (Fisher Scientific, Pittsburgh, PA). NADPH, fexofenadine, digoxin, and prazosin were purchased from Sigma–Aldrich Research (St. Louis, MO). Pooled human liver microsomes, probe P450 substrates (phenacetin, diclofenac, S-mephenytoin,
Cytochrome P450 Inhibition by CP-100,356
The ability of CP-100,356 to function as a competitive inhibitor of the five major human P450 enzymes namely P4501A2, 2C9, 2C19, 2D6, and 3A4 was examined in human liver microsomes. Considering that P4503A4 inhibition is dependent on substrate used in the microsomal assay,35 two structurally distinct probe substrates namely testosterone and midazolam were chosen for analysis of P4503A4 inhibition by CP-100,356. Virtually no inhibition (IC50 > 50 µM) of the catalytic activity of the individual
DISCUSSION
An initial literature survey for selecting MDR1 inhibitors in this work was not encouraging considering that many inhibitors possessed additional “off-target” activity including inhibition of P4503A4 and/or other transport proteins like MRP2 and OATP1B1 which are also known to be involved in active efflux or influx.18., 19., 20., 21., 22., 23. Therefore, we focused our attention on the diaminoquinazoline derivative CP-100,356, an optimized lead compound developed in-house as a MDR1 inhibitor.31
Acknowledgements
The authors thank Jonathan Bauman for technical assistance in the pharmacokinetic studies.
REFERENCES (64)
Mechanisms and strategies to overcome chemotherapy resistance in metastatic breast cancer
Cancer Treat Rev
(2008)- et al.
The role of efflux pumps in drug-resistant metastatic breast cancer: New insights and treatment strategies
Clin Breast Cancer
(2007) - et al.
In vivo use of the P450 inactivator 1-aminobenzotriazole in the rat: Varied dosing route to elucidate gut and liver contributions to first-pass and systemic clearance
J Pharm Sci
(2006) - et al.
Cyclosporine interacts with mycophenolic acid by inhibiting the multidrug resistance-associated protein 2
Am J Transplant
(2005) - et al.
Substrate specificity for rat cytochrome P450 (CYP) isoforms: Screening with cDNA-expressed systems of the rat
Biochem Pharmacol
(2002) - et al.
Relative contribution of absorptive and secretory transporters to the intestinal absorption of fexofenadine in rats
Int J Pharm
(2008) - et al.
Effect of fruit juices on the oral bioavailability of fexofenadine in rats
J Pharm Sci
(2005) - et al.
Cloning and characterization of a member of the rat multidrug resistance (mdr) gene family
Gene
(1991) - et al.
Mammalian ABC transporters in health and human disease
Annu Rev Biochem
(2002) - et al.
Role of transport proteins in drug absorption, distribution and excretion
Xenobiotica
(2001)
Role of P-glyoprotein in pharmacokinetics
Clin Pharmacokinet
Impact of drug transporter studies on drug discovery and development
Pharmacol Rev
Transporters and drug discovery: Why, when, and how
Mol Pharm
Breast cancer resistance protein (Bcrp/abcg2) is a major determinant of sulfasalazine absorption and elimination in the mouse
Mol Pharm
P-glycoprotein and breast cancer resistance protein: Two dominant transporters working together in limiting the brain penetration of topotecan
Clin Cancer Res
The effect of P-gp (Mdr1a/1b), BCRP (Bcrp1) and P-gp/BCRP inhibitors on the in vivo absorption, distribution, metabolism and excretion of imatinib
Invest New Drugs
The role of efflux and uptake transporters in[N-{3-chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methylsulfonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamine (GW572016, lapatinib) disposition and drug interactions
Drug Metab Dispos
Concise review: Clinical relevance of drug drug and herb drug interactions mediated by the ABC transporter ABCB1 (MDR1, P-glycoprotein)
Oncologist
Breast cancer resistance protein in pharmacokinetics and drug-drug interactions
Expert Opin Drug Metab Toxicol
Role of P-glycoprotein in statin drug interactions
Pharmacotherapy
Drug interactions with lipid-lowering drugs: Mechanisms and clinical relevance
Clin Pharmacol Ther
Application of three-dimensional quantitative structure-activity relationships of P-glycoprotein inhibitors and substrates
Mol Pharmacol
Characterization of substrates and inhibitors for the in vitro assessment of Bcrp mediated drug-drug interactions
Pharm Res
Glycoprotein and cytochrome P-450 3A inhibition: Dissociation of inhibitory potencies
Cancer Res
Mibefradil is a P-glycoprotein substrate and a potent inhibitor of both P-glycoprotein and CYP3A in vitro
Drug Metab Dispos
HIV-1 protease inhibitors are substrates for the MDR1 multidrug transporter
Biochemistry
Substantial pharmacokinetic interaction between digoxin and ritonavir in healthy volunteers
Clin Pharmacol Ther
In vivo effects of cyclosporin A and ketoconazole on the pharmacokinetics of representative substrates for P-glycoprotein and cytochrome P450 (CYP) 3A in rats
Biol Pharm Bull
Effects of ketoconazole on the intestinal metabolism, transport and oral bioavailability of K02, a novel vinylsulfone peptidomimetic cysteine protease inhibitor and a P450 3A, P-glycoprotein dual substrate, in male Sprague-Dawley rats
J Pharmacol Exp Ther
First-pass metabolism of midazolam by the human intestine
Clin Pharmacol Ther
Structurally distinct MDR modulators show specific patterns of reversal against p-glycoproteins bearing unique mutations at serine939/941
Biochemistry
Human (MDR1) and Mouse (mdr1,mdr3) p-glycoproteins can be distinguished by their respective drug resistance profiles and sensitivity to modulators
Biochemistry
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