Article Figures & Data
Figures
- Fig. 1.
ABC1183 selectively inhibits GSK3 α/β and CDK9. (A) Structure of ABC1183. (B) Top, in vitro kinase screen with 10 μM ABC1183 and graphical representation of inhibition. Bottom, four kinases with inhibition >60%. (C) Dose-response assay calculating IC50 values of GSK3α, GSK3β, and CDK9.
- Fig. 2.
ABC1183 exhibits cytotoxicity against a panel of murine and human cancer cells. (A) The indicated cell lines were seeded into 96-well plates at determined subconfluent concentrations and 24 hours later were treated with a dose response of ABC1183 for 72 hours. Cell survival was quantified by sulforhodamine B assay. IC50 value for each cell line shown. (B) Pan02 cells were treated with dimethyl sulfoxide (DMSO) or 3 μM ABC1183 for 24 hours and cell cycle was analyzed by propidium iodide staining. Representative histogram plots are shown.
- Fig. 3.
Multiple tumorigenic signaling pathways are inhibited by ABC1183. (A) LNCaP, (B) Pan02, (C) FaDu cells were treated with 3 μM ABC1183 for 2–24 hours or 20 nM LY2090314, as indicated. Cell lysates were immunoblotted with MCL1, pSer21/9 GSK3α/β, GSK3β, pSer641 GS, GS, pSer33/37 Thr41 β-catenin, β-catenin, and GAPDH.
- Fig. 4.
Inhibition of tumor growth by ABC1183. C57BL/6 mice were injected subcutaneously with (A) B16 melanoma cells suspended in PBS, (B) PAN02 pancreatic cancer cells suspended in PBS/growth matrix, or (C) TRAMP-C2 murine prostate cancer cells suspended in PBS/growth matrix. After tumor volume of at least 100 mm3, animals were treated five times per week by oral gavage of either 100 μl vehicle (open squares), 5 mg/kg ABC1183 (A and B), or 2 mg/kg ABC1183 (C) (gray squares). Tumor volume measurement was performed for time course as indicated; n = 5 per group in (A) and (C), n = 10 per group for (B). *P < 0.05; **P < 0.01. (D) Pan02-bearing mice were treated with 5 or 25 mg/kg for 2 or 12 hours. Tumors were excised, snap frozen, processed, and tumor lysates immunoblotted with pSer21/9 GSK3α/β, GSK3β, pSer641 GS, and GS.
- Fig. 5.
ABC1183 is safe for oral administration and reduces severity of TNBS-colitis model, at least in part, through inflammatory cytokines. (A) Seven-day toxicity of ABC1183. The indicated dose of ABC1183 was administered by oral gavage daily for 7 days, and the mice were then sacrificed for hematology, blood chemistry, and organ weight analyses. Values are mean ± S.E.M., n = 3 or 4 mice/group. (B) C57BL/6 mice were treated with rectal 50% ethanol and oral vehicle (black bars); rectal TNBS and oral vehicle (cross hatched bars), or rectal TNBS or oral ABC1183 (gray bars 50 mg/kg). Animals were sacrificed on day 10 and macroscopic inflammation within the distal 3 cm of each colon was scored. (C) Mice were treated as in (B) and TNF-α and IL-6 cytokines levels were determined from colon samples. *P < 0.05; **P < 0.01.
- Fig. 6.
Effect of oral ABC1183 on disease parameters in the mouse DSS-induced ulcerative colitis. C57BL/6 mice were treated with oral vehicle (black bars), 2% DSS-positive control (horizontal hatched bars), or 2% DSS with oral ABC1183 50 mg/kg twice a day (gray bars), and the following parameters were measured: (A) Disease Activity Index (DAI), which monitors disease severity; (B) colon length on day 6 of treatment; (C) myeloperoxidase (MPO) activity measured from the colons of animals on day 6 of treatment. (D) Cytokine values: TNFα, IL-6, and IL-10 cytokine levels were determined from colon samples. *P < 0.05; **P < 0.01.
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Dual Targeting GSK3 and CDK9 with Novel ABC1183
