A novel ligand of the translationally controlled tumor protein (TCTP) identified by virtual drug screening for cancer differentiation therapy

Abstract

SummaryIntroduction Differentiation therapy is a promising strategy for cancer treatment. The translationally controlled tumor protein (TCTP) is an encouraging target in this context. By now, this field of research is still at its infancy, which motivated us to perform a large-scale screening for the identification of novel ligands of TCTP. We studied the binding mode and the effect of TCTP blockade on the cell cycle in different cancer cell lines. Methods Based on the ZINC-database, we performed virtual screening of 2,556,750 compounds to analyze the binding of small molecules to TCTP. The in silico results were confirmed by microscale thermophoresis. The effect of the new ligand molecules was investigated on cancer cell survival, flow cytometric cell cycle analysis and protein expression by Western blotting and co-immunoprecipitation in MOLT-4, MDA-MB-231, SK-OV-3 and MCF-7 cells. Results Large-scale virtual screening by PyRx combined with molecular docking by AutoDock4 revealed five candidate compounds. By microscale thermophoresis, ZINC10157406 (6-(4-fluorophenyl)-2-[(8-methoxy-4-methyl-2-quinazolinyl)amino]-4(3H)-pyrimidinone) was identified as TCTP ligand with a KD of 0.87 ± 0.38. ZINC10157406 revealed growth inhibitory effects and caused G0/G1 cell cycle arrest in MOLT-4, SK-OV-3 and MCF-7 cells. ZINC10157406 (2 × IC50) downregulated TCTP expression by 86.70 ± 0.44% and upregulated p53 expression by 177.60 ± 12.46%. We validated ZINC10157406 binding to the p53 interaction site of TCTP and replacing p53 by co-immunoprecipitation. Discussion ZINC10157406 was identified as potent ligand of TCTP by in silico and in vitro methods. The compound bound to TCTP with a considerably higher affinity compared to artesunate as known TCTP inhibitor. We were able to demonstrate the effect of TCTP blockade at the p53 binding site, i.e. expression of TCTP decreased, whereas p53 expression increased. This effect was accompanied by a dose-dependent decrease of CDK2, CDK4, CDK, cyclin D1 and cyclin D3 causing a G0/G1 cell cycle arrest in MOLT-4, SK-OV-3 and MCF-7 cells. Our findings are supposed to stimulate further research on TCTP-specific small molecules for differentiation therapy in oncology.

Studying molecular interactions via capillary electrophoresis and microscale thermophoresis: A review

“…Recently, Fischer et al. [187] discovered that new inhibitors based on in silico drug screening for about 2556 750 lead compounds via using of ZINC database and molecular docking with certain criteria. Five leads have been selected to study the inhibition effect against translationally controlled tumor protein (TCTP).…”

Disease-Ligand Identification Based on Flexible Neural Tree

“…( Morris et al, 1996 ; Kellenberger et al, 2004 ; Taufer et al, 2005 ). Fischer et al (2021) utilized virtual screening method to screen 25, 56, 750 compounds in order to make the analysis about the binding of small molecules to translationally controlled tumor protein. Baxter et al (2000) utilized molecular docking to screen ligand-receptor complexes in virtual database and tabu search method was utilized to assist this work.…”

Revisiting the Molecular Interactions between the Tumor Protein TCTP and the Drugs Sertraline/Thioridazine

“…The accumulating knowledge on TCTP has made it an attractive target for therapeutic strategies in cancers [2,3,4,5,6,7,33,34,35]. Indeed its overexpression in many tumor cell types [34] confers tumor cells with increased survival and chemoresistance, and TCTP depletion by silencing approaches or by small molecules render tumor cells more sensitive to apoptosis and treatment, thus reducing oncogenic traits [18,33,34,36,37,38,39,40,41,42,43,44]. The most promising approach to target TCTP in cancer was introduced by the group of A. Telerman and R. Amson [34].…”

“…[2][3][4][5][6][7][34][35][36] Indeed its overexpression in many tumor cell types [35] confers tumor cells with increased survival and chemoresistance, and TCTP depletion by silencing approaches or by small molecules render tumor cells more sensitive to apoptosis and treatment, thus reducing oncogenic traits. [19,34,35,[37][38][39][40][41][42][43][44][45] The most promising approach to target TCTP in cancer was introduced by the group of A. Telerman and R. Amson. [35] They showed the growth inhibitory effect of anti-histaminic drugs hydroxyzine and promethazine on U937 cells in the 10 μM concentration range which was accompanied by reduced TCTP intracellular levels.…”