Identification of the first highly selective inhibitor of human GABA transporter GAT3

TY - JOUR

T1 - Identification of the first highly selective inhibitor of human GABA transporter GAT3

AU - Damgaard, Maria

AU - Al-Khawaja, Anas

AU - Vogensen, Stine B.

AU - Jurik, A

AU - Sijm, M

AU - Lie, M

AU - Bæk, M.I.

AU - Rosenthal, Emil

AU - Jensen, Anders A.

AU - Ecker, G.F.

AU - Frølund, Bente

AU - Wellendorph, Petrine

AU - Clausen, Rasmus P.

PY - 2015/7/8

Y1 - 2015/7/8

N2 - Screening a library of small-molecule compounds using a cell line expressing human GABA transporter 3 (hGAT3) in a [(3)H]GABA uptake assay identified isatin derivatives as a new class of hGAT3 inhibitors. A subsequent structure-activity relationship (SAR) study led to the identification of hGAT3-selective inhibitors (i.e., compounds 20 and 34) that were superior to the reference hGAT3 inhibitor, (S)-SNAP-5114, in terms of potency (low micromolar IC50 values) and selectivity (>30-fold selective for hGAT3 over hGAT1/hGAT2/hBGT1). Further pharmacological characterization of compound 20 (5-(thiophen-2-yl)indoline-2,3-dione) revealed a noncompetitive mode of inhibition at hGAT3. This suggests that this compound class, which has no structural resemblance to GABA, has a binding site different from the substrate, GABA. This was supported by a molecular modeling study that suggested a unique binding site that matched the observed selectivity, inhibition kinetics, and SAR of the compound series. These compounds are the most potent GAT3 inhibitors reported to date that provide selectivity for GAT3 over other GABA transporter subtypes.

AB - Screening a library of small-molecule compounds using a cell line expressing human GABA transporter 3 (hGAT3) in a [(3)H]GABA uptake assay identified isatin derivatives as a new class of hGAT3 inhibitors. A subsequent structure-activity relationship (SAR) study led to the identification of hGAT3-selective inhibitors (i.e., compounds 20 and 34) that were superior to the reference hGAT3 inhibitor, (S)-SNAP-5114, in terms of potency (low micromolar IC50 values) and selectivity (>30-fold selective for hGAT3 over hGAT1/hGAT2/hBGT1). Further pharmacological characterization of compound 20 (5-(thiophen-2-yl)indoline-2,3-dione) revealed a noncompetitive mode of inhibition at hGAT3. This suggests that this compound class, which has no structural resemblance to GABA, has a binding site different from the substrate, GABA. This was supported by a molecular modeling study that suggested a unique binding site that matched the observed selectivity, inhibition kinetics, and SAR of the compound series. These compounds are the most potent GAT3 inhibitors reported to date that provide selectivity for GAT3 over other GABA transporter subtypes.

U2 - 10.1021/acschemneuro.5b00150

DO - 10.1021/acschemneuro.5b00150

M3 - Journal article

SN - 1948-7193

VL - 6

SP - 1591

EP - 1599

JO - A C S Chemical Neuroscience

JF - A C S Chemical Neuroscience

ER -