Studies of the biogenic amine transporters. XI. Identification of a 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine (GBR12909) analog that allosterically modulates the serotonin transporter

Barbara Nightingale, Christina M. Dersch, Terrence L. Boos, Elisabeth Greiner, William Calhoun, Arthur E. Jacobson, Kenner C. Rice, Richard B. Rothman

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Previous studies identified partial inhibitors of serotonin (5-HT) transporter and dopamine transporter binding. We report here on a partial inhibitor of 5-HT transporter (SERT) binding identified among a group of 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine analogs (4-[2-[bis(4-fluorophenyl)methoxy]ethyl]-1-(2-trifluoromethyl-benzyl) -piperidine; TB-1-099). Membranes were prepared from rat brains or human embryonic kidney cells expressing the cloned human dopamine (hDAT), serotonin (hSERT), and norepinephrine (hNET) transporters. β-(4′- 125Iodophenyl)tropan-2β-carboxylic acid methyl ester ([ 125I]RTI-55) binding and other assays followed published procedures. Using rat brain membranes, TB-1-099 weakly inhibited DAT binding (Ki = 439 nM), was inactive at NET binding ([3H]nisoxetine), and partially inhibited SERT binding with an extrapolated plateau ("A" value) of 20%. Similarly, TB-1-099 partially inhibited [125I]RTI-55 binding to hSERT with an extrapolated plateau (A value) of 14%. Upon examining the effect of increasing concentrations of TB-1-099 on the apparent K d and Bmax of [125I]RTI-55 binding to hSERT, we found that TB-1-099 decreased the Bmax in a dose-dependent manner and affected the apparent Kd in a manner well described by a sigmoid dose-response curve. TB-1-099 increased the Kd but not to the magnitude expected for a competitive inhibitor. In rat brain synaptosomes, TB-1-099 noncompetitively inhibited [3H]5-HT, but not [ 3H]dopamine, uptake. Dissociation experiments indicated that TB-1-099 promoted the rapid dissociation of a small component of [125I]RTI-55 binding to hSERT. Association experiments demonstrated that TB-1-099 slowed [125I]RTI-55 binding to hSERT in a manner unlike that of the competitive inhibitor indatraline. Viewed collectively, these results support the hypothesis that TB-1-099 allosterically modulates hSERT binding and function.

Original languageEnglish (US)
Pages (from-to)906-915
Number of pages10
JournalJournal of Pharmacology and Experimental Therapeutics
Volume314
Issue number2
DOIs
StatePublished - Aug 2005
Externally publishedYes

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Serotonin Plasma Membrane Transport Proteins
Biogenic Amines
Serotonin
nisoxetine
Dopamine
Brain
4-(2-(bis(4-fluorophenyl)methoxy)ethyl)-1-(2-trifluoromethylbenzyl)piperidine
vanoxerine
Norepinephrine Plasma Membrane Transport Proteins
Dopamine Plasma Membrane Transport Proteins
Membranes
Synaptosomes
Sigmoid Colon
Carboxylic Acids
Esters
RTI 55
Kidney

ASJC Scopus subject areas

  • Pharmacology

Cite this

Studies of the biogenic amine transporters. XI. Identification of a 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine (GBR12909) analog that allosterically modulates the serotonin transporter. / Nightingale, Barbara; Dersch, Christina M.; Boos, Terrence L.; Greiner, Elisabeth; Calhoun, William; Jacobson, Arthur E.; Rice, Kenner C.; Rothman, Richard B.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 314, No. 2, 08.2005, p. 906-915.

Research output: Contribution to journalArticle

Nightingale, Barbara ; Dersch, Christina M. ; Boos, Terrence L. ; Greiner, Elisabeth ; Calhoun, William ; Jacobson, Arthur E. ; Rice, Kenner C. ; Rothman, Richard B. / Studies of the biogenic amine transporters. XI. Identification of a 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine (GBR12909) analog that allosterically modulates the serotonin transporter. In: Journal of Pharmacology and Experimental Therapeutics. 2005 ; Vol. 314, No. 2. pp. 906-915.
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abstract = "Previous studies identified partial inhibitors of serotonin (5-HT) transporter and dopamine transporter binding. We report here on a partial inhibitor of 5-HT transporter (SERT) binding identified among a group of 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine analogs (4-[2-[bis(4-fluorophenyl)methoxy]ethyl]-1-(2-trifluoromethyl-benzyl) -piperidine; TB-1-099). Membranes were prepared from rat brains or human embryonic kidney cells expressing the cloned human dopamine (hDAT), serotonin (hSERT), and norepinephrine (hNET) transporters. β-(4′- 125Iodophenyl)tropan-2β-carboxylic acid methyl ester ([ 125I]RTI-55) binding and other assays followed published procedures. Using rat brain membranes, TB-1-099 weakly inhibited DAT binding (Ki = 439 nM), was inactive at NET binding ([3H]nisoxetine), and partially inhibited SERT binding with an extrapolated plateau ({"}A{"} value) of 20{\%}. Similarly, TB-1-099 partially inhibited [125I]RTI-55 binding to hSERT with an extrapolated plateau (A value) of 14{\%}. Upon examining the effect of increasing concentrations of TB-1-099 on the apparent K d and Bmax of [125I]RTI-55 binding to hSERT, we found that TB-1-099 decreased the Bmax in a dose-dependent manner and affected the apparent Kd in a manner well described by a sigmoid dose-response curve. TB-1-099 increased the Kd but not to the magnitude expected for a competitive inhibitor. In rat brain synaptosomes, TB-1-099 noncompetitively inhibited [3H]5-HT, but not [ 3H]dopamine, uptake. Dissociation experiments indicated that TB-1-099 promoted the rapid dissociation of a small component of [125I]RTI-55 binding to hSERT. Association experiments demonstrated that TB-1-099 slowed [125I]RTI-55 binding to hSERT in a manner unlike that of the competitive inhibitor indatraline. Viewed collectively, these results support the hypothesis that TB-1-099 allosterically modulates hSERT binding and function.",
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AU - Nightingale, Barbara

AU - Dersch, Christina M.

AU - Boos, Terrence L.

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AU - Rice, Kenner C.

AU - Rothman, Richard B.

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N2 - Previous studies identified partial inhibitors of serotonin (5-HT) transporter and dopamine transporter binding. We report here on a partial inhibitor of 5-HT transporter (SERT) binding identified among a group of 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine analogs (4-[2-[bis(4-fluorophenyl)methoxy]ethyl]-1-(2-trifluoromethyl-benzyl) -piperidine; TB-1-099). Membranes were prepared from rat brains or human embryonic kidney cells expressing the cloned human dopamine (hDAT), serotonin (hSERT), and norepinephrine (hNET) transporters. β-(4′- 125Iodophenyl)tropan-2β-carboxylic acid methyl ester ([ 125I]RTI-55) binding and other assays followed published procedures. Using rat brain membranes, TB-1-099 weakly inhibited DAT binding (Ki = 439 nM), was inactive at NET binding ([3H]nisoxetine), and partially inhibited SERT binding with an extrapolated plateau ("A" value) of 20%. Similarly, TB-1-099 partially inhibited [125I]RTI-55 binding to hSERT with an extrapolated plateau (A value) of 14%. Upon examining the effect of increasing concentrations of TB-1-099 on the apparent K d and Bmax of [125I]RTI-55 binding to hSERT, we found that TB-1-099 decreased the Bmax in a dose-dependent manner and affected the apparent Kd in a manner well described by a sigmoid dose-response curve. TB-1-099 increased the Kd but not to the magnitude expected for a competitive inhibitor. In rat brain synaptosomes, TB-1-099 noncompetitively inhibited [3H]5-HT, but not [ 3H]dopamine, uptake. Dissociation experiments indicated that TB-1-099 promoted the rapid dissociation of a small component of [125I]RTI-55 binding to hSERT. Association experiments demonstrated that TB-1-099 slowed [125I]RTI-55 binding to hSERT in a manner unlike that of the competitive inhibitor indatraline. Viewed collectively, these results support the hypothesis that TB-1-099 allosterically modulates hSERT binding and function.

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