Cirtuvivint

Cell lines

SW480 cells

Preparation Method

SW480 cells (7500 cells/well) were incubated overnight, then treated with DMSO, Cirtuvivint (5 doses, range 0.03μM-3μM), or staurosporine (0.1μM) at 37°C for 48h. Cells were then incubated with CellEvent™ caspase 3/7 green detection reagent at 37°C for 30min, followed by Hoechst 33342 (blue) dye. Quantification of apoptotic (green-labeled) and total (blue-labeled nuclei) cells per well was performed on a CellInsight™ CX5 imager for all wells of three replicate plates, and the ratio of green:blue cells was calculated and expressed as a percentage; the result was the average of the triplicate wells.

Reaction Conditions

0.03μM-3μM; 48h

Applications

Cirtuvivint induced cell apoptosis. Treatment of SW480 cells with Cirtuvivint resulted in increased levels of activated caspase 3/7.

Animal models

Athymic nude mice (Charles River Labs, Strain 553)

Preparation Method

Mice were subcutaneously injected with 5×10⁶ HEC265, Ishikawa, or Ishikawa-S33Y cells, or 1×10⁷ HEC265 or SNGM cells on the right flank. Tumor progression was measured using calipers every 2 days, and tumor volume as well as % change in tumor volume were recorded. Once a tumor in each flank population grew to over 100mm³, treatment was initiated in all mice. For all experiments, mice were treated with paclitaxel alone (10mg/kg weekly, i.p.), Cirtuvivint alone (12.5mg/kg, daily, p.o.), paclitaxel+Cirtuvivint, or vehicle. Body weight was measured twice per week during treatment as a surrogate for toxicity. Body weight loss of >10% was considered indicative of severe reaction to treatment. The mice were sacrificed, and the tumors were surgically resected and weighed. Tumor burden was calculated based on the weight of resected tumors. Blood samples were collected at time of death via cardiac punction for complete blood count (CBC) and blood chemistry analysis (AST, ALT, and ALP), blood volume permitting.

Dosage form

12.5mg/kg/day; p.o.

Applications

The efficacy of Cirtuvivint alone is comparable to that of paclitaxel alone. The combination therapy of Cirtuvivint and paclitaxel is more effective than either drug used alone.

References:
[1] Tam B Y, Chiu K, Chung H, et al. The CLK inhibitor SM08502 induces anti-tumor activity and reduces Wnt pathway gene expression in gastrointestinal cancer models[J]. Cancer letters, 2020, 473: 186-197.
[2]Corr B R, Moroney M R, Woodruff E, et al. Combination CDC-like kinase inhibition (CLK)/Dual-specificity tyrosine-regulated kinase (DYRK) and taxane therapy in CTNNB1-mutated endometrial cancer[J]. bioRxiv, 2023.