AG-012986 is a pan- CDK inhibitor with K i values of 44 nM ( CDK1 ), 9.2 nM ( CDK4 ), 94 nM ( CDK2 ), and IC50 values of 22 nM ( CDK5 ), 4 nM ( CDK9 ). AG-012986 causes apoptosis of T-cells by targeting upstream kinases in the p38 Mitogen-activated protein kinase ( MAPK ) pathway and impairing cellular survival. AG-012986 induces cell cycle arrest, retinoblastoma protein hypophosphorylation, and reduces K i -67 expression. AG-012986 exerts antiproliferative activity in tumor cells, demonstrates antitumor efficacy in human xenograft models, and causes retinal and peripheral neurotoxicity, plus immune cell toxicity. AG-012986 can be used for the research of colon carcinoma, non-small cell lung carcinoma, lung carcinoma, breast carcinoma, ovarian tumor, pancreatic carcinoma, osteosarcoma, lymphoma, leukemia, retinotoxicity [1] [2] [3] [4].
In Vivo, AG-012986 (8.8-40 mg/kg; s.c.; daily, 12 days) shows >83% tumor growth inhibition (TGI) in 10 of the 11 tumor models tested[1].
In Vitro, AG-012986 is a potent, selective pan-CDK inhibitor with nanomolar activity against CDK4/cyclin D3 (Ki = 9.2 nM), CDK1/cyclin B (Ki = 44 nM), CDK2/cyclin A (Ki = 94 nM), CDK9/cyclin T (IC50 = 4 nM), and CDK5/p35 (IC50 = 22 nM)[1]. AG-012986 (0.837-2.44 μM) has limited off-target activity against non-Kinase targets, with functional interactions at micromolar concentrations with the calcium type L ion channel, serotonin transporter, and histamine H3 receptor[1]. AG-012986 (72 h) potently inhibits proliferation of 18 human tumor cell lines with an average IC50 of 120 nM, and displays IC50 values <100 nM in 13 of these cell lines, independent of p53 and Rb status[1]. AG-012986 (60-240 nM; 8-24 h) induces dose-dependent hypophosphorylation of Rb Ser795 in HCT116 human colon cancer cells after 24 hours of treatment, with maximal effects at >120 nM, but shows minimal effect after 8 hours[1]. AG-012986 (30 nM-1 μM; 8-24 h) induces G1 phase arrest in HCT116 human colon cancer cells at 30 to 120 nM and G2-M phase arrest at ≥240 nM after 24 hours of treatment, but does not induce cell cycle arrest with <8 hours of exposure[1]. AG-012986 (30-240 nM; 8-24 h) induces apoptosis in HCT116 human colon cancer cells with an IC50 of ≈160 nM after 24 hours of treatment, but shows no apoptotic effect after 8 hours[1]. AG-012986 (10-1000 nM; 8-320 h) shows time-dependent cytotoxicity in SW620 human colon carcinoma cells, with minimal activity after 8 hours, moderate activity (IC50 = 300 nM) after 24 hours, and substantial cytotoxicity (IC50 <100 nM) after ≥72 hours[1]. AG-012986 exhibits binding inhibition of multiple CDK isoforms in a cell-free KINOMEscan assay, with similar IC50 values to non-neurotoxic NVP-2 for CDK11, CDK16, and CDK17[2]. AG-012986 (200-500 nM; 24 h) induces dose-dependent cytotoxicity in human MIO-M1 Müller cells, with significant decreases in cell viability and increases in cell death observed at 200 nM and maximal effects at 500 nM after 24 h of incubation[2]. AG-012986 (250 nM; 16 h) induces rapid apoptosis/necrosis in primary human PBMCs, with 80% of cells showing dual PI/Annexin-V positivity[3]. AG-012986 (50 nM-1 μM; 8-48 h) activates caspase-3/7 in primary human PBMCs in vitro in a dose-dependent manner, with peak activity occurring 16-24 h after treatment with 50 nM, 200 nM, or 1 μM[3]. AG-012986 (50 nM-200 nM; 8 h) induces cleavage of caspase-3 and PARP in primary human PBMCs following 8 h treatment with 50 nM or 200 nM, confirming caspase-mediated apoptosis[3]. AG-012986 (10 nM-500 nM; 16 h) exhibits greater cytotoxicity in primary human PBMCs than staurosporine after 16 h treatment with 10-500 nM, as measured by reduced ATP content and increased caspase-3/7 activity[3]. AG-012986's (50 nM-250 nM; 20 min) toxicity profile is altered by acute stimulation of T cells[3]. AG-012986 (20 nM-500 nM) inhibits α-CD3-induced proliferation in purified primary human T cells pretreated by AG-012986 followed by 24 h α-CD3 antibody stimulation, while α-CD3 stimulation decouples this antiproliferative activity from AG-012986-induced toxicity[3]. AG-012986 (20 nM-500 nM; 48 h) inhibits α-CD3-induced IL-2 production in purified primary human T cells in a dose-dependent manner, with a fivefold reduction observed at 20 nM[3]. AG-012986 (50-250 nM; 20 min pretreatment) inhibits both basal and α-CD3-induced p38 phosphorylation in purified primary human T cells pretreated with 50 nM or 250 nM for 20 min followed by 16 h incubation with or without stimulation[3]. AG-012986 (250 nM; 16 h) causes rapidly apoptosis in PBMCs and occurs independently of any cell division[4]. AG-012986 (50-200 nM; 8 h) induced the presence of p17 and p19 and the cleaved form of PARP[4].
References:
[1]. Zhang C, et al. Pharmacologic properties of AG-012986, a pan-cyclin-dependent kinase inhibitor with antitumor efficacy. Mol Cancer Ther. 2008 Apr;7(4):818-28.
[2]. Wright P, et al. Differential expression of cyclin-dependent kinases in the adult human retina in relation to CDK inhibitor retinotoxicity. Arch Toxicol. 2019;93(3):659-671.
[3]. Lee DU, et al. Off-target immune cell toxicity caused by AG-012986, a pan-CDK inhibitor, is associated with inhibition of p38 MAPK phosphorylation. J Biochem Mol Toxicol. 2012 Mar;26(3):101-8.
In Vivo, AG-012986 (8.8-40 mg/kg; s.c.; daily, 12 days) shows >83% tumor growth inhibition (TGI) in 10 of the 11 tumor models tested[1].
In Vitro, AG-012986 is a potent, selective pan-CDK inhibitor with nanomolar activity against CDK4/cyclin D3 (Ki = 9.2 nM), CDK1/cyclin B (Ki = 44 nM), CDK2/cyclin A (Ki = 94 nM), CDK9/cyclin T (IC50 = 4 nM), and CDK5/p35 (IC50 = 22 nM)[1]. AG-012986 (0.837-2.44 μM) has limited off-target activity against non-Kinase targets, with functional interactions at micromolar concentrations with the calcium type L ion channel, serotonin transporter, and histamine H3 receptor[1]. AG-012986 (72 h) potently inhibits proliferation of 18 human tumor cell lines with an average IC50 of 120 nM, and displays IC50 values <100 nM in 13 of these cell lines, independent of p53 and Rb status[1]. AG-012986 (60-240 nM; 8-24 h) induces dose-dependent hypophosphorylation of Rb Ser795 in HCT116 human colon cancer cells after 24 hours of treatment, with maximal effects at >120 nM, but shows minimal effect after 8 hours[1]. AG-012986 (30 nM-1 μM; 8-24 h) induces G1 phase arrest in HCT116 human colon cancer cells at 30 to 120 nM and G2-M phase arrest at ≥240 nM after 24 hours of treatment, but does not induce cell cycle arrest with <8 hours of exposure[1]. AG-012986 (30-240 nM; 8-24 h) induces apoptosis in HCT116 human colon cancer cells with an IC50 of ≈160 nM after 24 hours of treatment, but shows no apoptotic effect after 8 hours[1]. AG-012986 (10-1000 nM; 8-320 h) shows time-dependent cytotoxicity in SW620 human colon carcinoma cells, with minimal activity after 8 hours, moderate activity (IC50 = 300 nM) after 24 hours, and substantial cytotoxicity (IC50 <100 nM) after ≥72 hours[1]. AG-012986 exhibits binding inhibition of multiple CDK isoforms in a cell-free KINOMEscan assay, with similar IC50 values to non-neurotoxic NVP-2 for CDK11, CDK16, and CDK17[2]. AG-012986 (200-500 nM; 24 h) induces dose-dependent cytotoxicity in human MIO-M1 Müller cells, with significant decreases in cell viability and increases in cell death observed at 200 nM and maximal effects at 500 nM after 24 h of incubation[2]. AG-012986 (250 nM; 16 h) induces rapid apoptosis/necrosis in primary human PBMCs, with 80% of cells showing dual PI/Annexin-V positivity[3]. AG-012986 (50 nM-1 μM; 8-48 h) activates caspase-3/7 in primary human PBMCs in vitro in a dose-dependent manner, with peak activity occurring 16-24 h after treatment with 50 nM, 200 nM, or 1 μM[3]. AG-012986 (50 nM-200 nM; 8 h) induces cleavage of caspase-3 and PARP in primary human PBMCs following 8 h treatment with 50 nM or 200 nM, confirming caspase-mediated apoptosis[3]. AG-012986 (10 nM-500 nM; 16 h) exhibits greater cytotoxicity in primary human PBMCs than staurosporine after 16 h treatment with 10-500 nM, as measured by reduced ATP content and increased caspase-3/7 activity[3]. AG-012986's (50 nM-250 nM; 20 min) toxicity profile is altered by acute stimulation of T cells[3]. AG-012986 (20 nM-500 nM) inhibits α-CD3-induced proliferation in purified primary human T cells pretreated by AG-012986 followed by 24 h α-CD3 antibody stimulation, while α-CD3 stimulation decouples this antiproliferative activity from AG-012986-induced toxicity[3]. AG-012986 (20 nM-500 nM; 48 h) inhibits α-CD3-induced IL-2 production in purified primary human T cells in a dose-dependent manner, with a fivefold reduction observed at 20 nM[3]. AG-012986 (50-250 nM; 20 min pretreatment) inhibits both basal and α-CD3-induced p38 phosphorylation in purified primary human T cells pretreated with 50 nM or 250 nM for 20 min followed by 16 h incubation with or without stimulation[3]. AG-012986 (250 nM; 16 h) causes rapidly apoptosis in PBMCs and occurs independently of any cell division[4]. AG-012986 (50-200 nM; 8 h) induced the presence of p17 and p19 and the cleaved form of PARP[4].
References:
[1]. Zhang C, et al. Pharmacologic properties of AG-012986, a pan-cyclin-dependent kinase inhibitor with antitumor efficacy. Mol Cancer Ther. 2008 Apr;7(4):818-28.
[2]. Wright P, et al. Differential expression of cyclin-dependent kinases in the adult human retina in relation to CDK inhibitor retinotoxicity. Arch Toxicol. 2019;93(3):659-671.
[3]. Lee DU, et al. Off-target immune cell toxicity caused by AG-012986, a pan-CDK inhibitor, is associated with inhibition of p38 MAPK phosphorylation. J Biochem Mol Toxicol. 2012 Mar;26(3):101-8.