Reversible dimers of the atypical antipsychotic quetiapine inhibit p-glycoprotein-mediated efflux in vitro with increased binding affinity and in situ at the blood-brain barrier

Dana Emmert, Christopher R. Campos, David Ward, Peihua Lu, Hilda A. Namanja, Kelsey Bohn, David S. Miller, Frances J. Sharom, Jean Chmielewski, Christine A. Hrycyna

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The multidrug resistance transporter P-glycoprotein (P-gp) is highly expressed in the capillary endothelial cells of the blood-brain barrier (BBB) where it functions to limit the brain penetration of many drugs, including antipsychotic agents used to treat schizophrenia. Therefore, in an effort to inhibit the transporter, we designed dimers of the antipsychotic drug and P-gp substrate quetiapine (QT), linked by variable length tethers. In P-gp overexpressing cells and in human brain capillary endothelial hCMEC/D3 cells, the dimer with the shortest tether length (QT2C2) (1) was the most potent inhibitor showing >80-fold better inhibition of P-gp-mediated transport than monomeric QT. The dimers, which are linked via ester moieties, are designed to revert to the therapeutic monomer once inside the target cells. We demonstrated that the addition of two sterically blocking methyl groups to the linker (QT2C2Me2, 8) increased the half-life of the molecule in plasma 10-fold as compared to the dimer lacking methyl groups (QT2C2, 1), while retaining inhibitory potency for P-gp transport and sensitivity to cellular esterases. Experiments with purified P-gp demonstrated that QT2C2 (1) and QT 2C2Me2 (8) interacted with both the H- and R-binding sites of the transporter with binding affinities 20- to 30-fold higher than that of monomeric QT. Using isolated rat brain capillaries, QT 2C2Me2 (8) was a more potent inhibitor of P-gp transport than QT. Lastly, we showed that QT2C2Me 2 (8) increased the accumulation of the P-gp substrate verapamil in rat brain in situ three times more than QT. Together, these results indicate that the QT dimer QT2C2Me2 (8) strongly inhibited P-gp transport activity in human brain capillary endothelial cells, in rat brain capillaries, and at the BBB in an animal model.

Original languageEnglish (US)
Pages (from-to)305-317
Number of pages13
JournalACS Chemical Neuroscience
Volume5
Issue number4
DOIs
StatePublished - Apr 16 2014
Externally publishedYes

Fingerprint

P-Glycoprotein
Blood-Brain Barrier
Dimers
Antipsychotic Agents
Glycoproteins
Brain
Rats
Endothelial cells
Endothelial Cells
Quetiapine Fumarate
In Vitro Techniques
Multiple Drug Resistance
Substrates
Esterases
Verapamil
Human Activities
Half-Life
Schizophrenia
Esters
Animals

Keywords

  • ABC transporter
  • antipsychotic agent
  • blood-brain barrier
  • inhibitor
  • P-glycoprotein
  • quetiapine

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Physiology
  • Cognitive Neuroscience
  • Medicine(all)

Cite this

Reversible dimers of the atypical antipsychotic quetiapine inhibit p-glycoprotein-mediated efflux in vitro with increased binding affinity and in situ at the blood-brain barrier. / Emmert, Dana; Campos, Christopher R.; Ward, David; Lu, Peihua; Namanja, Hilda A.; Bohn, Kelsey; Miller, David S.; Sharom, Frances J.; Chmielewski, Jean; Hrycyna, Christine A.

In: ACS Chemical Neuroscience, Vol. 5, No. 4, 16.04.2014, p. 305-317.

Research output: Contribution to journalArticle

Emmert, Dana ; Campos, Christopher R. ; Ward, David ; Lu, Peihua ; Namanja, Hilda A. ; Bohn, Kelsey ; Miller, David S. ; Sharom, Frances J. ; Chmielewski, Jean ; Hrycyna, Christine A. / Reversible dimers of the atypical antipsychotic quetiapine inhibit p-glycoprotein-mediated efflux in vitro with increased binding affinity and in situ at the blood-brain barrier. In: ACS Chemical Neuroscience. 2014 ; Vol. 5, No. 4. pp. 305-317.
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abstract = "The multidrug resistance transporter P-glycoprotein (P-gp) is highly expressed in the capillary endothelial cells of the blood-brain barrier (BBB) where it functions to limit the brain penetration of many drugs, including antipsychotic agents used to treat schizophrenia. Therefore, in an effort to inhibit the transporter, we designed dimers of the antipsychotic drug and P-gp substrate quetiapine (QT), linked by variable length tethers. In P-gp overexpressing cells and in human brain capillary endothelial hCMEC/D3 cells, the dimer with the shortest tether length (QT2C2) (1) was the most potent inhibitor showing >80-fold better inhibition of P-gp-mediated transport than monomeric QT. The dimers, which are linked via ester moieties, are designed to revert to the therapeutic monomer once inside the target cells. We demonstrated that the addition of two sterically blocking methyl groups to the linker (QT2C2Me2, 8) increased the half-life of the molecule in plasma 10-fold as compared to the dimer lacking methyl groups (QT2C2, 1), while retaining inhibitory potency for P-gp transport and sensitivity to cellular esterases. Experiments with purified P-gp demonstrated that QT2C2 (1) and QT 2C2Me2 (8) interacted with both the H- and R-binding sites of the transporter with binding affinities 20- to 30-fold higher than that of monomeric QT. Using isolated rat brain capillaries, QT 2C2Me2 (8) was a more potent inhibitor of P-gp transport than QT. Lastly, we showed that QT2C2Me 2 (8) increased the accumulation of the P-gp substrate verapamil in rat brain in situ three times more than QT. Together, these results indicate that the QT dimer QT2C2Me2 (8) strongly inhibited P-gp transport activity in human brain capillary endothelial cells, in rat brain capillaries, and at the BBB in an animal model.",
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AU - Lu, Peihua

AU - Namanja, Hilda A.

AU - Bohn, Kelsey

AU - Miller, David S.

AU - Sharom, Frances J.

AU - Chmielewski, Jean

AU - Hrycyna, Christine A.

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