Female
athymic CrTac:NCr-Foxn1nu mice from Taconic (6–8 weeks of age) were injected subcutaneously
in the flank with 3 million COLO205-F1756 clone 4 cells, and when
tumor xenografts reached a mean diameter of 5–6 mm (equating
to a mean volume of about 100 mm3), mice (n = 8 per group) were dosed orally by gavage with compound 74 (70 mg/kg bid for 8 days) or vehicle (10% dimethyl sulfoxide; 5%
Tween 20 in sterile saline). Luminescent flux as a measure of WNT
pathway activity was quantified by whole body imaging in cohorts of
four animals at 2 and 6 h after the final dose. This was achieved
by sc injection of 100 μL of 30 mg/mL D-Luciferin firefly substrate
(XR-1001, PerkinElmer) into anaesthetized mice and imaging with a
Xenogen IVIS 200 (XR-1001, PerkinElmer), utilizing the software Living
Image (version 4). At these time points, heparinized blood samples
were collected by cardiac puncture under terminal anesthesia, and
plasma was separated by centrifugation. Tumors were excised and weighed,
and both plasma and tumor samples were snap frozen for PK analyses.
Mallinger A., Crumpler S., Pichowicz M., Waalboer D., Stubbs M., Adeniji-Popoola O., Wood B., Smith E., Thai C., Henley A.T., Georgi K., Court W., Hobbs S., Box G., Ortiz-Ruiz M.J., Valenti M., De Haven Brandon A., TePoele R., Leuthner B., Workman P., Aherne W., Poeschke O., Dale T., Wienke D., Esdar C., Rohdich F., Raynaud F., Clarke P.A., Eccles S.A., Stieber F., Schiemann K, & Blagg J. (2015). Discovery of Potent, Orally Bioavailable, Small-Molecule Inhibitors of WNT Signaling from a Cell-Based Pathway Screen. Journal of Medicinal Chemistry, 58(4), 1717-1735.