GPS167 is a CLK kinase inhibitor that potently and selectively inhibits recombinant human CLK1, CLK2 and CLK4. By inhibiting CLK-mediated phosphorylation of SRSF10, GPS167 upregulates the protein-binding ability of CLK1 and CLK4 with SRSF10, downregulates oncogenic BCLAF1-L and upregulates tumor-suppressive BCLAF1-S, regulates alternative splicing of genes such as MDM2 and MDM4, stabilizes p53 protein and induces DNA damage, ultimately triggering tumor cell apoptosis. GPS167 can block the epithelial-mesenchymal transition process of tumors, activate intracellular double-stranded RNA-mediated antiviral immune responses, and produce synergistic cytotoxicity when combined with microtubule-targeting drugs. GPS167 can be used in research related to various cancers including colorectal cancer [1] [2] [3].
In Vitro, GPS167 potently and selectively inhibits recombinant human CLK1, CLK2 and CLK4 with IC50 values of 0.0896 μM, 0.0820 μM and 0.1230 μM, and shows weak or negligible activity against most other tested human kinases. In non-radioactive proximity assays, GPS167 yields an IC50 of 0.5069 μM for CLK1 and displays poor inhibitory potency toward DYRK1A and DYRK1B; and its inhibitory effect on SRSF10 phosphorylation is preferential than its inhibitory effect on SRSF1 phosphorylation[1][2]. GPS167 (1-30 μM; 6 h) increases Thr212-Tau phosphorylation in SH-SY5Y cells expressing human 4R-Tau[1]. GPS167 (48 h) is non-cytotoxic to HT22 murine hippocampal neuronal cells, with an IC50 for viability >100 μM[1]. GPS167 (0.05-100 μM; 48 h) modulates BCLAF1 alternative splicing in HCT116 cells, reducing BCLAF1-L variant levels with an EC50 of ~2 μM[2]. GPS167 (10-20 μM; 24 h cell incubation, 10 min pull-down incubation) promotes dephosphorylation of specific serine residues on SRSF10 and increases the interaction between SRSF10 and CLK1 in HCT116 cells[2]. GPS167 (10 μM; 0-80 h) induces potent cytotoxicity in human colorectal cancer cell lines (HCT116, COLO205, SW620) but has minimal effects on Caco-2 cells and normal colonocyte cell lines (CRL-1831, CRL-1790)[2]. GPS167 induces apoptosis in human colorectal cancer HCT116, COLO205 and SW620 cells accompanied by elevated cleaved PARP and cleaved caspase-3, whereas no apoptotic effect is observed in Caco-2 and normal colon epithelial CRL-1831 cells. It inhibits cancer cell migration as evidenced by decreased scratch wound closure rate[2]. GPS167 (10-20 μM; up to 120 h viability, up to 84 h cytotoxicity, 72 h PARP cleavage) impairs viability, induces cytotoxicity and apoptosis, and disrupts structure in human and mouse colorectal cancer organoids, but has minimal effects on normal colorectal organoids[2]. GPS167 (10-20 μM; 24 h viability and caspase activity, up to 72 h cytotoxicity) elicits p53-dependent growth inhibition, cytotoxicity, and apoptosis in HCT116 colorectal cancer cells, with increased p53 expression and DNA damage observed in p53-proficient cells[2]. GPS167 (8 days) in NALM-6 cells, depletion of genes involved in mitotic cell cycle checkpoint/chromosome segregation increases sensitivity to GPS167, while depletion of mitotic spindle assembly-related genes reduces GPS167 sensitivity[3]. GPS167 disrupts the expression and alternative splicing of EMT-relevant genes, including FLNB and CD44, in HCT116 colorectal cancer cells[3]. GPS167 (1 μM; 4 days) in MCF10A breast epithelial cells, upregulates the epithelial marker ECAD (both with and without TGF-β treatment) and inhibits TGF-β-induced EMT morphological changes, without altering mesenchymal marker expression[3]. GPS167 (20 μM; up to 72 h) in HCT116 colorectal cancer cells, depletion of the dsRNA sensor DHX33 significantly reduces cytotoxicity over 72 hours[3]. GPS167 inhibits proliferation and induces p53-dependent apoptosis in multiple cancer cell lines (including colorectal, melanoma, leukemia, breast, ovarian, and renal) and colorectal organoids, while sparing normal colonocyte cells and organoids[3].
References:
[1]. Lindberg MF, et al. Comparative Efficacy and Selectivity of Pharmacological Inhibitors of DYRK and CLK Protein Kinases. J Med Chem. 2023;66(6):4106-4130.
[2]. Sohail M, et al. A novel class of inhibitors that target SRSF10 and promote p53-mediated cytotoxicity on human colorectal cancer cells. NAR Cancer. 2021;3(2):zcab019. Published 2021 May 25.
[3]. Shkreta L, et al. The anticancer potential of the CLK kinases inhibitors 1C8 and GPS167 revealed by their impact on the epithelial-mesenchymal transition and the antiviral immune response. Oncotarget. 2024 May 16;15:313-325.
In Vitro, GPS167 potently and selectively inhibits recombinant human CLK1, CLK2 and CLK4 with IC50 values of 0.0896 μM, 0.0820 μM and 0.1230 μM, and shows weak or negligible activity against most other tested human kinases. In non-radioactive proximity assays, GPS167 yields an IC50 of 0.5069 μM for CLK1 and displays poor inhibitory potency toward DYRK1A and DYRK1B; and its inhibitory effect on SRSF10 phosphorylation is preferential than its inhibitory effect on SRSF1 phosphorylation[1][2]. GPS167 (1-30 μM; 6 h) increases Thr212-Tau phosphorylation in SH-SY5Y cells expressing human 4R-Tau[1]. GPS167 (48 h) is non-cytotoxic to HT22 murine hippocampal neuronal cells, with an IC50 for viability >100 μM[1]. GPS167 (0.05-100 μM; 48 h) modulates BCLAF1 alternative splicing in HCT116 cells, reducing BCLAF1-L variant levels with an EC50 of ~2 μM[2]. GPS167 (10-20 μM; 24 h cell incubation, 10 min pull-down incubation) promotes dephosphorylation of specific serine residues on SRSF10 and increases the interaction between SRSF10 and CLK1 in HCT116 cells[2]. GPS167 (10 μM; 0-80 h) induces potent cytotoxicity in human colorectal cancer cell lines (HCT116, COLO205, SW620) but has minimal effects on Caco-2 cells and normal colonocyte cell lines (CRL-1831, CRL-1790)[2]. GPS167 induces apoptosis in human colorectal cancer HCT116, COLO205 and SW620 cells accompanied by elevated cleaved PARP and cleaved caspase-3, whereas no apoptotic effect is observed in Caco-2 and normal colon epithelial CRL-1831 cells. It inhibits cancer cell migration as evidenced by decreased scratch wound closure rate[2]. GPS167 (10-20 μM; up to 120 h viability, up to 84 h cytotoxicity, 72 h PARP cleavage) impairs viability, induces cytotoxicity and apoptosis, and disrupts structure in human and mouse colorectal cancer organoids, but has minimal effects on normal colorectal organoids[2]. GPS167 (10-20 μM; 24 h viability and caspase activity, up to 72 h cytotoxicity) elicits p53-dependent growth inhibition, cytotoxicity, and apoptosis in HCT116 colorectal cancer cells, with increased p53 expression and DNA damage observed in p53-proficient cells[2]. GPS167 (8 days) in NALM-6 cells, depletion of genes involved in mitotic cell cycle checkpoint/chromosome segregation increases sensitivity to GPS167, while depletion of mitotic spindle assembly-related genes reduces GPS167 sensitivity[3]. GPS167 disrupts the expression and alternative splicing of EMT-relevant genes, including FLNB and CD44, in HCT116 colorectal cancer cells[3]. GPS167 (1 μM; 4 days) in MCF10A breast epithelial cells, upregulates the epithelial marker ECAD (both with and without TGF-β treatment) and inhibits TGF-β-induced EMT morphological changes, without altering mesenchymal marker expression[3]. GPS167 (20 μM; up to 72 h) in HCT116 colorectal cancer cells, depletion of the dsRNA sensor DHX33 significantly reduces cytotoxicity over 72 hours[3]. GPS167 inhibits proliferation and induces p53-dependent apoptosis in multiple cancer cell lines (including colorectal, melanoma, leukemia, breast, ovarian, and renal) and colorectal organoids, while sparing normal colonocyte cells and organoids[3].
References:
[1]. Lindberg MF, et al. Comparative Efficacy and Selectivity of Pharmacological Inhibitors of DYRK and CLK Protein Kinases. J Med Chem. 2023;66(6):4106-4130.
[2]. Sohail M, et al. A novel class of inhibitors that target SRSF10 and promote p53-mediated cytotoxicity on human colorectal cancer cells. NAR Cancer. 2021;3(2):zcab019. Published 2021 May 25.
[3]. Shkreta L, et al. The anticancer potential of the CLK kinases inhibitors 1C8 and GPS167 revealed by their impact on the epithelial-mesenchymal transition and the antiviral immune response. Oncotarget. 2024 May 16;15:313-325.