SPR-based fragment screening with neurotensin receptor 1 generates novel small molecule ligands

• Published in: PloS one 2017-05, Vol.12 (5), p.e0175842-e0175842
• Main Authors: Huber, Sylwia, Casagrande, Fabio, Hug, Melanie N, Wang, Lisha, Heine, Philipp, Kummer, Lutz, Plückthun, Andreas, Hennig, Michael
• Format: Article
• Language: English
• Subjects: Animal models; Antibodies; Antipsychotics; Biochemistry; Biological activity; Biology and Life Sciences; Blood-brain barrier; Brain research; CCR5 protein; Central nervous system; Chemokine receptors; Clinical trials; Cognitive ability; Computer Simulation; Crystal structure; Crystallization; Crystallography; CXCR4 protein; Data processing; Drug Discovery - methods; Drug Evaluation, Preclinical; Enzymes; Health aspects; Humans; Ion channels; Kinases; Kinetics; Ligands; Ligases; Lipids; Macromolecules; Magnetic resonance; Magnetic Resonance Spectroscopy; Medical screening; Membrane proteins; Membranes; Mental disorders; Methods; Models, Molecular; Molecular Conformation; Molecular weight; Neurotensin; NMR; Nuclear magnetic resonance; Peptides; Pharmacokinetics; Pharmacology; Physical Sciences; Protein Binding; Protein Stability; Proteins; Receptors, Neurotensin - agonists; Receptors, Neurotensin - antagonists & inhibitors; Receptors, Neurotensin - chemistry; Receptors, Neurotensin - metabolism; Reproducibility of Results; Research and Analysis Methods; Residues; Resonance; Sexually transmitted diseases; Small Molecule Libraries; Social Sciences; Spectroscopy; STD; Surface plasmon resonance; Technology; Workflow
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0175842

The neurotensin receptor 1 represents an important drug target involved in various diseases of the central nervous system. So far, the full exploitation of potential therapeutic activities has been compromised by the lack of compounds with favorable physicochemical and pharmacokinetic properties which efficiently penetrate the blood-brain barrier. Recent progress in the generation of stabilized variants of solubilized neurotensin receptor 1 and its subsequent purification and successful structure determination presents a solid starting point to apply the approach of fragment-based screening to extend the chemical space of known neurotensin receptor 1 ligands. In this report, surface plasmon resonance was used as primary method to screen 6369 compounds. Thereby 44 hits were identified and confirmed in competition as well as dose-response experiments. Furthermore, 4 out of 8 selected hits were validated using nuclear magnetic resonance spectroscopy as orthogonal biophysical method. Computational analysis of the compound structures, taking the known crystal structure of the endogenous peptide agonist into consideration, gave insight into the potential fragment-binding location and interactions and inspires chemistry efforts for further exploration of the fragments.