Iterative focused screening with biological fingerprints identifies selective Asc-1 inhibitors distinct from traditional high throughput screening

Kutchukian, P. S. et al. (2017) Iterative focused screening with biological fingerprints identifies selective Asc-1 inhibitors distinct from traditional high throughput screening. ACS Chemical Biology, 12(2), pp. 519-527. (doi: 10.1021/acschembio.6b00913) (PMID:28032990)

146530.pdf - Accepted Version



N-methyl-d-aspartate receptors (NMDARs) mediate glutamatergic signaling that is critical to cognitive processes in the central nervous system, and NMDAR hypofunction is thought to contribute to cognitive impairment observed in both schizophrenia and Alzheimer’s disease. One approach to enhance the function of NMDAR is to increase the concentration of an NMDAR coagonist, such as glycine or d-serine, in the synaptic cleft. Inhibition of alanine–serine–cysteine transporter-1 (Asc-1), the primary transporter of d-serine, is attractive because the transporter is localized to neurons in brain regions critical to cognitive function, including the hippocampus and cortical layers III and IV, and is colocalized with d-serine and NMDARs. To identify novel Asc-1 inhibitors, two different screening approaches were performed with whole-cell amino acid uptake in heterologous cells stably expressing human Asc-1: (1) a high-throughput screen (HTS) of 3 M compounds measuring 35S l-cysteine uptake into cells attached to scintillation proximity assay beads in a 1536 well format and (2) an iterative focused screen (IFS) of a 45 000 compound diversity set using a 3H d-serine uptake assay with a liquid scintillation plate reader in a 384 well format. Critically important for both screening approaches was the implementation of counter screens to remove nonspecific inhibitors of radioactive amino acid uptake. Furthermore, a 15 000 compound expansion step incorporating both on- and off-target data into chemical and biological fingerprint-based models for selection of additional hits enabled the identification of novel Asc-1-selective chemical matter from the IFS that was not identified in the full-collection HTS.

Item Type:Articles
Glasgow Author(s) Enlighten ID:Thomson, Dr Fiona
Authors: Kutchukian, P. S., Warren, L., Magliaro, B. C., Amoss, A., Cassaday, J. A., O’Donnell, G., Squadroni, B., Zuck, P., Pascarella, D., Culberson, J. C., Cooke, A. J., Hurzy, D., Schlegel, K.-A. S., Thomson, F., Johnson, E. N., Uebele, V. N., Hermes, J. D., Parmentier-Batteur, S., and Finley, M.
College/School:College of Medical Veterinary and Life Sciences > School of Cancer Sciences
Journal Name:ACS Chemical Biology
Publisher:American Chemical Society
ISSN (Online):1554-8937
Published Online:29 December 2016
Copyright Holders:Copyright © 2016 American Chemical Society
First Published:First published in ACS Chemical Biology 12(2):519-527
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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