Diphenhydramine as a selective probe to study H+-antiporter function at the blood–brain barrier: Application to 11Cdiphenhydramine positron emission tomography imaging

摘要

Diphenhydramine, a sedative histamine H1-receptor (H1R) antagonist, was evaluated as a probe to measure drug/H⁺-antiporter function at the blood–brain barrier. In situ brain perfusion experiments in mice and rats showed that diphenhydramine transport at the blood–brain barrier was saturable, following Michaelis–Menten kinetics with a Km = 2.99 mM and Vmax = 179.5 nmol s⁻¹ g⁻¹. In the pharmacological plasma concentration range the carrier-mediated component accounted for 77% of diphenhydramine influx while passive diffusion accounted for only 23%. [¹⁴C]Diphenhydramine blood–brain barrier transport was proton and clonidine sensitive but was influenced by neither tetraethylammonium, a MATE1 (SLC47A1), and OCT/OCTN (SLC22A1-5) modulator, nor P-gp/Bcrp (ABCB1a/1b/ABCG2) deficiency. Brain and plasma kinetics of [¹¹C]diphenhydramine were measured by positron emission tomography imaging in rats. [¹¹C]Diphenhydramine kinetics in different brain regions were not influenced by displacement with 1 mg kg⁻¹ unlabeled diphenhydramine, indicating the specificity of the brain positron emission tomography signal for blood–brain barrier transport activity over binding to any central nervous system target in vivo. [¹¹C]Diphenhydramine radiometabolites were not detected in the brain 15 min after injection, allowing for the reliable calculation of [¹¹C]diphenhydramine brain uptake clearance (Clup = 0.99 ± 0.18 mL min⁻¹cm⁻³). Diphenhydramine is a selective and specific H⁺-antiporter substrate. [¹¹C]Diphenhydramine positron emission tomography imaging offers a reliable and noninvasive method to evaluate H⁺-antiporter function at the blood–brain barrier.