Development and characterization of LLC-PK1 cells containing Sprague–Dawley rat Abcb1a (Mdr1a): Comparison of rat P-glycoprotein transport to human and mouse

Booth-Genthe, Catherine; Louie, Steven W.; Carlini, Edward J.; Li, Bing; Leake, Brenda F.; Eisenhandler, Roy; Hochman, Jerome H.; Qin, Mei; Kim, Richard B.; Rushmore, Thomas H.; Yamazaki, Masayo

doi:10.1016/j.vascn.2005.12.004

Cited by 30 publications

(25 citation statements)

References 32 publications

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“…Appropriate positive and negative controls were used in each assessment. The transcellular permeability with LLC-PK1 and efflux transport by P-glycoprotein (Pgp) and breast cancer resistance protein (BCRP) was assessed in MDR1-LLC-PK1 and BCRP-MDCKII cell monolayers (Schinkel et al, 1995;Booth-Genthe et al, 2006). The [ 14 C] AMG 416 direct uptake assay was conducted in HEK293 cells transfected with organic anion transporter (OAT) 1, OAT3, OAT polypeptide (OATP) 1B1, OATP1B3, peptide transporter 1, 2, or organic cation transporter (OCT) 2 transporter (Yamazaki et al, 2005;Chu et al, 2007).…”

Section: In Vitro Methodsmentioning

confidence: 99%

Nonclinical Pharmacokinetics, Disposition, and Drug-Drug Interaction Potential of a Novel D-Amino Acid Peptide Agonist of the Calcium-Sensing Receptor AMG 416 (Etelcalcetide)

Subramanian

Zhu

Kerr

et al. 2016

Drug Metabolism and Disposition

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AMG 416 (etelcalcetide) is a novel synthetic peptide agonist of the calcium-sensing receptor composed of a linear chain of seven D-amino acids (referred to as the D-amino acid backbone) with a D-cysteine linked to an L-cysteine via a disulfide bond. AMG 416 contains four basic D-arginine residues and is a +4 charged peptide at physiologic pH with a mol. wt. of 1048.3 Da. The pharmacokinetics (PK), disposition, and potential of AMG 416 to cause drug-drug interaction were investigated in nonclinical studies with two single 14 C-labels placed either at a potentially metabolically labile acetyl position or on the D-alanine next to D-cysteine in the interior of the D-amino acid backbone. After i.v. dosing, the PK and disposition of AMG 416 were similar in male and female rats. Radioactivity rapidly distributed to most tissues in rats with intact kidneys, and renal elimination was the predominant clearance pathway. No straindependent differences were observed. In bilaterally nephrectomized rats, minimal radioactivity (1.2%) was excreted via nonrenal pathways. Biotransformation occurred primarily via disulfide exchange with endogenous thiol-containing molecules in whole blood rather than metabolism by enzymes, such as proteases or cytochrome P450s; the D-amino acid backbone remained unaltered. A substantial proportion of the plasma radioactivity was covalently conjugated to albumin. AMG 416 presents a low risk for P450 or transportermediated drug-drug interactions because it showed no interactions in vitro. These studies demonstrated a 14 C label on either the acetyl or the D-alanine in the D-amino acid backbone would be appropriate for clinical studies.

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Section: In Vitro Methodsmentioning

confidence: 99%

Nonclinical Pharmacokinetics, Disposition, and Drug-Drug Interaction Potential of a Novel D-Amino Acid Peptide Agonist of the Calcium-Sensing Receptor AMG 416 (Etelcalcetide)

Subramanian

Zhu

Kerr

et al. 2016

Drug Metabolism and Disposition

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“…Directional transport studies to determine P-gp transport ratios were performed according to published methods (Yamazaki et. al, 2001;Booth-Genthe et al, 2006) using Multiscreen Caco-2 96-well filter plates (Millipore Corporation, Billerica, MA). Passive membrane permeability values were determined as the average of the permeability coefficients for apical to basolateral (A-B) and basolateral to apical (B-A) transport in the parental LLC-PK1 cells, whereas the P-glycoprotein ratio is measured as the ratio of B-A/A-B permeation.…”

Section: Methodsmentioning

confidence: 99%

First Demonstration of Cerebrospinal Fluid and Plasma Aβ Lowering with Oral Administration of a β-Site Amyloid Precursor Protein-Cleaving Enzyme 1 Inhibitor in Nonhuman Primates

Sankaranarayanan¹,

Holahan²,

Colussi³

et al. 2008

J Pharmacol Exp Ther

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103

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␤-Site amyloid precursor protein (APP)-cleaving enzyme (BACE) 1 cleavage of amyloid precursor protein is an essential step in the generation of the potentially neurotoxic and amyloidogenic A␤42 peptides in Alzheimer's disease. Although previous mouse studies have shown brain A␤ lowering after BACE1 inhibition, extension of such studies to nonhuman primates or man was precluded by poor potency, brain penetration, and pharmacokinetics of available inhibitors. In this study, a novel tertiary carbinamine BACE1 inhibitor, tertiary carbinamine (TC)-1, was assessed in a unique cisterna magna ported rhesus monkey model, where the temporal dynamics of A␤ in cerebrospinal fluid (CSF) and plasma could be evaluated. TC-1, a potent inhibitor (IC 50 ϳ 0.4 nM), has excellent passive membrane permeability, low susceptibility to P-glycoprotein transport, and lowered brain A␤ levels in a mouse model. Intravenous infusion of TC-1 led to a significant but transient lowering of CSF and plasma A␤ levels in conscious rhesus monkeys because it underwent CYP3A4-mediated metabolism. Oral codosing of TC-1 with ritonavir, a potent CYP3A4 inhibitor, twice daily over 3.5 days in rhesus monkeys led to sustained plasma TC-1 exposure and a significant and sustained reduction in CSF sAPP␤, A␤40, A␤42, and plasma A␤40 levels. CSF A␤42 lowering showed an EC 50 of ϳ20 nM with respect to the CSF [TC-1] levels, demonstrating excellent concordance with its potency in a cell-based assay. These results demonstrate the first in vivo proof of concept of CSF A␤ lowering after oral administration of a BACE1 inhibitor in a nonhuman primate.

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“…Studies investigating differences in P-gp functionality between rats and mice have primarily been conducted in vitro, applying efflux ratios derived from bidirectional transport studies in cell systems transfected with mdr1 from each species. These studies suggest some correlation between rat and mouse P-gp-mediated efflux and also differences, depending on the substrate (Booth-Genthe et al, 2006;Katoh et al, 2006;. However, comparison of efflux ratios among species from in vitro transport studies is circumvented with indecisive biological challenges such as total protein expression and density of the functional transporter in each cell line.…”

Section: Introductionmentioning

confidence: 99%

Species Comparison of In Vivo P-Glycoprotein-Mediated Brain Efflux Using mdr1a-Deficient Rats and Mice

Bundgaard¹,

Jensen²,

Garmer³

2011

Drug Metab Dispos

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ABSTRACT:The experiments described herein compared the extent of in vivo P-glycoprotein (P-gp)-mediated brain efflux between rats and mice for a set of known central nervous system compounds. With use of newly introduced genetically modified mdr1a-deficient rats and their gene-competent counterparts, the brain to plasma distribution was assessed and compared with the distribution pattern in mdr1a-deficient and wild-type mice. Four compounds (aripiprazole, citalopram, risperidone, and venlafaxine) were administered using a continuous subcutaneous osmotic minipump infusion paradigm. Steady-state brain and plasma concentrations of the compounds, including selected metabolites (9-hydroxyrisperidone, O-desmethyl-venlafaxine and N-desmethyl-venlafaxine) were measured in mdr1a-deficient rats and mice and their wild-type counterparts along with their free fractions to determine total and unbound brain to plasma distribution between genotypes within and between species. The results revealed qualitative as well as quantitative similarities between P-gp functionality in vivo at the blood-brain barrier level in rats and mice. All compounds tested were shown to have a significantly higher brain to plasma distribution in both mdr1a-deficient rats and mice compared with that in their wild-type counterparts. Moreover, the relative enhancement in extent of brain penetration between mdr1a-deficient and wild-type rats could be directly correlated to the enhancement ratios obtained in mice. From the unbound brain to unbound plasma distributions, the impact of P-gp on the overall brain penetration capabilities showed minor differences between rats and mice for the compounds tested. In conclusion, a comparable functional role of P-gp between rats and mice with respect to brain efflux mediated by this transporter is suggested.

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Development and characterization of LLC-PK1 cells containing Sprague–Dawley rat Abcb1a (Mdr1a): Comparison of rat P-glycoprotein transport to human and mouse

Cited by 30 publications

References 32 publications

Nonclinical Pharmacokinetics, Disposition, and Drug-Drug Interaction Potential of a Novel D-Amino Acid Peptide Agonist of the Calcium-Sensing Receptor AMG 416 (Etelcalcetide)

Nonclinical Pharmacokinetics, Disposition, and Drug-Drug Interaction Potential of a Novel D-Amino Acid Peptide Agonist of the Calcium-Sensing Receptor AMG 416 (Etelcalcetide)

First Demonstration of Cerebrospinal Fluid and Plasma Aβ Lowering with Oral Administration of a β-Site Amyloid Precursor Protein-Cleaving Enzyme 1 Inhibitor in Nonhuman Primates

Species Comparison of In Vivo P-Glycoprotein-Mediated Brain Efflux Using mdr1a-Deficient Rats and Mice

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