MS‐Based Screening of 5‐Substituted Nipecotic Acid Derived Hydrazone Libraries as Ligands of the GABA Transporter 1

A screening of compound libraries based on nipecotic acid derivatives with lipophilic residues attached to the scarcely explored 5‐position of the core structure was used for the search of new inhibitors of the γ‐aminobutyric acid (GABA) transporter 1 (mGAT1). The generated compound libraries, which were based on hydrazone chemistry commonly used in dynamic combinatorial chemistry but rendered pseudostatic, were screened for their binding affinities toward mGAT1 by means of MS Binding Assays. With nipecotic acid derived hydrazone rac‐16 h [rac‐(3R,5S)‐{5‐[(E)‐2‐{[5‐(2‐phenylethynyl)thiophen‐2‐yl]methylidene}hydrazin‐1‐yl]piperidine‐3‐carboxylic acid}‐sodium chloride (1/2)], one hit was found and evaluated displaying sub‐micromolar potency (pKi=6.62±0.04) and a noncompetitive interaction mode at mGAT1. By bearing a 5‐(2‐phenylethynyl)thiophen‐2‐yl residue attached to the 5‐position of nipecotic acid via a three‐atom spacer, compound rac‐16 h contains a structural moiety so far unprecedented for these kinds of bioactive molecules, and complements novel 5‐substituted nipecotic acid derived ligands of mGAT1 revealed in a recently published screening campaign. This new class of ligands, with an inhibition mode distinct from that of benchmark mGAT1 inhibitors, could serve as research tools for investigations of mGAT1‐mediated GABA transport. On the search for new GABA uptake inhibitors: An MS‐based screening of pseudostatic dynamic combinatorial hydrazone libraries revealed a new ligand of the GABA transporter mGAT1, representing a 5‐substituted nipecotic acid derivative. Hit verification by means of MS Binding Assays indicated sub‐micromolar potency and a noncompetitive interaction mode at mGAT1 for the compound synthesized in pure form.