High-Throughput Screening Reveals New Glutaminase Inhibitor Molecules

Author(s):Show less -

Costa, Renna K. E. ^{[1, 2]} ; Rodrigues, Camila T. ^[3] ; Campos, Jean C. H. ^[2] ; Paradela, Luciana S. ^{[1, 2]} ; Dias, Marilia M. ^[2] ; da Silva, Bianca Novaes ^[2] ; de Valega Negrao, Cyro von Zuben ^{[1, 2]} ; Goncalves, Kaliandra de Almeida ^[2] ; Ascencao, Carolline F. R. ^{[1, 2]} ; Adamoski, Douglas ^{[1, 2]} ; Mercaldi, Gustavo Fernando ^[2] ; Bastos, Alliny C. S. ^[2] ; Batista, Fernanda A. H. ^[2] ; Figueira, Ana Carolina ^[2] ; Cordeiro, Artur T. ^[2] ; Ambrosio, Andre L. B. ^[2] ; Guido, Rafael V. C. ^[3] ; Dias, Sandra M. G. ^[2]

Affiliation

^[1] Univ Campinas UNICAMP, Inst Biol, Grad Program Genet & Mol Biol, BR-13083970 Campinas, SP - Brazil

^[2] Ctr Res Energy & Mat CNPEM, Brazilian Biosci Natl Lab LNBio, BR-13083100 Campinas, SP - Brazil

^[3] Univ Sao Paulo, Sao Carlos Inst Phys IFSC, BR-13563120 Sao Carlos, SP - Brazil

• Document type: Journal article
• Source: ACS PHARMACOLOGY & TRANSLATIONAL SCIENCE; v. 4, n. 6, p. 1849-1866, DEC 10 2021.
• Web of Science Citations: 0

Abstract

The glutaminase (GLS) enzyme hydrolyzes glutamine into glutamate, an important anaplerotic source for the tricarboxylic acid cycle in rapidly growing cancer cells under the Warburg effect. Glutamine-derived alpha-ketoglutarate is also an important cofactor of chromatin-modifying enzymes, and through epigenetic changes, it keeps cancer cells in an undifferentiated state. Moreover, glutamate is an important neurotransmitter, and deregulated glutaminase activity in the nervous system underlies several neurological disorders. Given the proven importance of glutaminase for critical diseases, we describe the development of a new coupled enzyme-based fluorescent glutaminase activity assay formatted for 384-well plates for high-throughput screening (HTS) of glutaminase inhibitors. We applied the new methodology to screen a similar to 30,000-compound library to search for GLS inhibitors. The HTS assay identified 11 glutaminase inhibitors as hits that were characterized by in silico, biochemical, and glutaminase-based cellular assays. A structure-activity relationship study on the most promising hit (C9) allowed the discovery of a derivative, C9.22, with enhanced in vitro and cellular glutaminase-inhibiting activity. In summary, we discovered a new glutaminase inhibitor with an innovative structural scaffold and described the molecular determinants of its activity. (AU)

• FAPESP's process: 13/23510-4 - Understanding how PI3K/AKt/mTOR and AMPK signaling pathways affect glutaminase activity.
• Grantee: Carolline Fernanda Rodrigues Ascenção
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 15/25832-4 - Metabolic regulation of genetic and epigenetic control of gene expression
• Grantee: Sandra Martha Gomes Dias
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/09077-4 - Developing new glutaminase inhibitors with potential anti-tumor activity
• Grantee: Renna Karoline Eloi Costa
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 13/07600-3 - CIBFar - Center for Innovation in Biodiversity and Drug Discovery
• Grantee: Glaucius Oliva
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 19/16351-3 - The glutaminase role on tumor progression
• Grantee: Sandra Martha Gomes Dias
• Support Opportunities: Regular Research Grants
• FAPESP's process: 14/17820-3 - Post-transcriptional regulation of glutaminase enzyme by HuR and its relationship with high glutaminolytic levels in tumors
• Grantee: Douglas Adamoski Meira
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 14/15968-3 - Understanding the glutaminase functional regulation and the development of inhibitors as new approaches to cancer therapy
• Grantee: Sandra Martha Gomes Dias
• Support Opportunities: Regular Research Grants