In vitro activity: NSC12 inhibits FGF-dependent tumor growth, angiogenesis, and metastases. NSC12 does not affect FGF2/heparin interaction, whereas it inhibits the binding of FGF2 to the immobilized receptor (ID50 ~30 μM). NSC12 interferes with FGF2/FGFR1 interaction without affecting the ability of the growth factor to interact with heparin or HSPGs. NSC12 also binds immobilized FGF3, FGF4, FGF6, FGF8, FGF16, FGF18, FGF20, and FGF22 with Kd values ranging between ~16 and ~120 μM. NSC12 may act as a multi-FGF trap by interacting with all members of the canonical FGF subfamilies. NSC12 hampers FGF23-mediated FGFR1 activation in Klotho-expressing Chinese hamster ovary (CHO) cells. Treatment with NSC12 causes the reduction of the S phase of the cell cycle in all tumor cell lines but LLC cells, in which an accumulation in the S phase is observed. NSC12 inhibits FGFR1, FGFR2, FGFR3, and FGFR4 phosphorylation in CHO cell transfectants. NSC12 inhibits the proliferation of various FGF-dependent murine and human cancer cell lines with no inhibitory effect on HCC827 cancer cells that harbor a tumor-driving mutation of the EGFR TK domain and on FGF-independent cancer cell lines.

Cell Assay: KATO Ⅲ cells are plated at 104 cells/well in 96 well-plates in RPMI medium plus 1% FBS. After 24 hr cells are treated with different FGFs (30 ng/ml) in the absence or presence of an optimal dose of NSC12 (1.0 or 3.0 μM) or NSC21. After 72 hr the MTT assay is performed according to manufacturer’s instructions. The optical density (OD) is determined using a plate reader at a test wavelength of 595 nm and a reference wavelength of 630 nm.

Cancer Cell. 2015 Aug 10;28(2):225-39.