10 research outputs found
Role of Hepatic Organic Anion Transporter 2 in the Pharmacokinetics of <i>R</i>- and <i>S</i>‑Warfarin: In Vitro Studies and Mechanistic Evaluation
Interindividual
variability in warfarin dose requirement demands personalized medicine
approaches to balance its therapeutic benefits (anticoagulation) and
bleeding risk. Cytochrome P450 2C9 (<i>CYP2C9</i>) genotype-guided
warfarin dosing is recommended in the clinic, given the more potent <i>S</i>-warfarin is primarily metabolized by CYP2C9. However,
only about 20–30% of interpatient variability in <i>S</i>-warfarin clearance is associated with <i>CYP2C9</i> genotype.
We evaluated the role of hepatic uptake in the clearance of <i>R</i>- and <i>S</i>-warfarin. Using stably transfected
HEK293 cells, both enantiomers were found to be substrates of organic
anion transporter (OAT)2 with a Michaelis–Menten constant (<i>K</i><sub>m</sub>) of ∼7–12 μM but did not
show substrate affinity for other major hepatic uptake transporters.
Uptake of both enantiomers by primary human hepatocytes was saturable
(<i>K</i><sub>m</sub> ≈ 7–10 μM) and
inhibitable by OAT2 inhibitors (e.g., ketoprofen) but not by OATP1B1/1B3
inhibitors (e.g., cyclosporine). To further evaluate the potential
role of hepatic uptake in <i>R</i>- and <i>S</i>-warfarin pharmacokinetics, mechanistic modeling and simulations
were conducted. A “bottom-up” PBPK model, developed
assuming that OAT2–CYPs interplay, well recovered clinical
pharmacokinetics, drug–drug interactions, and <i>CYP2C9</i> pharmacogenomics of <i>R</i>- and <i>S</i>-warfarin.
Clinical data were not available to directly verify the impact of
OAT2 modulation on warfarin pharmacokinetics; however, the bottom-up
PBPK model simulations suggested a proportional change in clearance
of both warfarin enantiomers with inhibition of OAT2 activity. These
results suggest that variable hepatic OAT2 function, in conjunction
with CYP2C, may contribute to the high population variability in warfarin
pharmacokinetics and possibly anticoagulation end points and thus
warrant further clinical investigation
The Human Specific CCR1 Antagonist CP-481,715 Inhibits Cell Infiltration and Inflammatory Responses in Human CCR1 Transgenic Mice
Organic Anion Transporter 2–Mediated Hepatic Uptake Contributes to the Clearance of High-Permeability–Low-Molecular-Weight Acid and Zwitterion Drugs: Evaluation Using 25 Drugs
<i>In vitro</i> studies with two human organic anion transporters: OAT2 and OAT7
<p>1.Penciclovir, ganciclovir, creatinine, <i>para</i>-aminohippuric acid (PAH), ketoprofen, estrone 3-O-sulfate (E3S), dehydroepiandrosterone 3-O-sulfate (DHEAS) and cyclic guanosine monophosphate (cGMP) were screened as substrates of human liver organic anion transporters OAT2 and OAT7.</p> <p>2.For OAT7, high uptake ratios (versus mock transfected HEK293 cells) of 29.6 and 15.3 were obtained with E3S and DHEAS. Less robust uptake ratios (≤3.6) were evident with the other substrates. OAT2 (transcript variant 1, OAT2-tv1) presented high uptake ratios of 30, 13, ∼35, ∼25, 8.5 and 9 with cGMP, PAH, penciclovir, ganciclovir, creatinine and E3S, respectively. No uptake was observed with DHEAS.</p> <p>3.Although not a substrate of either transporter, ketoprofen did inhibit transfected OAT2-tv1 (IC<sub>50</sub> of 17, 22, 23, 24, 35 and 586 μM; creatinine, ganciclovir, penciclovir, cGMP, E3S and prostaglandin F2α, respectively) and penciclovir uptake (IC<sub>50</sub> = 27 µM; >90% inhibition) by plated human hepatocytes (PHH).</p> <p>4.It is concluded that penciclovir and ketoprofen may serve as useful tools for the assessment of OAT2 activity in PHH. However, measurement of OAT7 activity therein will prove more challenging, as high uptake rates are evident with E3S and DHEAS only and both sulfoconjugates are known to be substrates of organic anion transporting polypeptides.</p
Quantitative Contribution of Six Major Transporters to the Hepatic Uptake of Drugs: “SLC-Phenotyping” Using Primary Human Hepatocytes
Feasibility of Singlet Analysis for Ligand Binding Assays: a Retrospective Examination of Data Generated Using the Gyrolab Platform
Expression of rat I-TAC/CXCL11/SCYA11 during central nervous system inflammation: comparison with other CXCR3 ligands
© 2007 United States and Canadian Academy of PathologyThe chemokines are a large gene superfamily with critical roles in development and immunity. The chemokine receptor CXCR3 appears to play a major role in the trafficking of activated Th1 lymphocytes. There are at least three major ligands for CXCR3: mig/CXCL9, IP-10/CXCL10 and I-TAC/CXCL11, and of these three ligands, CXCL11 is the least well-characterized. In this study, we have cloned a rat ortholog of CXCL11, evaluated its function, and examined its expression in the Th-1-mediated disease, experimental autoimmune encephalomyelitis (EAE) in the rat. Based on its predicted primary amino-acid sequence, rat I-TAC/CXCL11 was synthesized and shown to induce chemotaxis of activated rat T lymphocytes in vitro and the in vivo migration of T lymphocytes when injected into the skin. I-TAC/CXCL11 expression, as determined by RT-PCR, increased in lymph node and spinal cord tissue collected from rats in which EAE had been actively induced, and in spinal cord tissue from rats in which EAE had been passively induced. The kinetics of expression were similar to that of CXCR3 and IP-10/CXCL10, although expression of both CXCR3 and IP-10/CXCL10 was more intense than that of I-TAC/CXCL11 and increased more rapidly in both lymph nodes and the spinal cord. Only minor levels of expression of the related chemokine mig/CXCL9 were observed. Immunohistochemistry revealed that the major cellular source of I-TAC/CXCL11 in the central nervous system (CNS) during EAE is likely to be the astrocyte. Together, these data indicate that I-TAC/CXCL11 is expressed in the CNS during the clinical phase of EAE. However, the observation that I-TAC/CXCL11 is expressed after receptor expression is detected suggests that it is not essential for the initial migration of CXCR3-bearing cells into the CNS.Shaun R McColl, Surendran Mahalingam, Maria Staykova, Laurie A Tylaska, Katherine E Fisher, Christine A Strick, Ronald P Gladue, Kuldeep S Neote and David O Willenbor
Discovery of Potent and Orally Bioavailable Macrocyclic Peptide–Peptoid Hybrid CXCR7 Modulators
The chemokine receptor CXCR7 is an
attractive target for a variety
of diseases. While several small-molecule modulators of CXCR7 have
been reported, peptidic macrocycles may provide advantages in terms
of potency, selectivity, and reduced off-target activity. We produced
a series of peptidic macrocycles that incorporate an N-linked peptoid
functionality where the peptoid group enabled us to explore side-chain
diversity well beyond that of natural amino acids. At the same time,
theoretical calculations and experimental assays were used to track
and reduce the polarity while closely monitoring the physicochemical
properties. This strategy led to the discovery of macrocyclic peptide–peptoid
hybrids with high CXCR7 binding affinities (<i>K</i><sub>i</sub> < 100 nM) and measurable passive permeability (<i>P</i><sub>app</sub> > 5 × 10<sup>–6</sup> cm/s).
Moreover, bioactive peptide <b>25</b> (<i>K</i><sub>i</sub> = 9 nM) achieved oral bioavailability of 18% in rats, which
was commensurate with the observed plasma clearance values upon intravenous
administration