title: CLM3, a multitarget tyrosine kinase inhibitor with antiangiogenic properties, is active against primary anaplastic thyroid cancer in vitro and in vivo.
abstract: OBJECTIVE
  We have studied the antitumor activity of a pyrazolo[3,4-d]pyrimidine compound (CLM3) proposed for a multiple signal transduction inhibition [including the RET tyrosine kinase, epidermal growth factor receptor, and vascular endothelial growth factor (VEGF) receptor and with antiangiogenic activity] in primary anaplastic thyroid cancer (ATC) cells, in the human cell line 8305C (undifferentiated thyroid cancer), and in an ATC-cell line (AF).
  
  
  METHODS
  CLM3 was tested in primary ATC cells at the concentrations of 5, 10, 30, and 50 μM; in 8305C cells, in AF cells, at 1, 5, 10, 30, 50, or 100 μM; and in AF cells in CD nu/nu mice.
  
  
  RESULTS
  CLM3 significantly inhibited the proliferation of 8305C and AF cells, also inducing apoptosis. A significant reduction of proliferation with CLM3 in ATC cells (P < .01, ANOVA) was shown. CLM3 increased the percentage of apoptotic ATC cells dose dependently (P < .001, ANOVA) and inhibited migration (P < .01) and invasion (P < .001). The AF cell line was injected sc in CD nu/nu mice, and tumor masses became detectable 15 days later. CLM3 (50 mg/kg per die) significantly inhibited tumor growth (starting 16 d after the beginning of treatment). CLM3 significantly decreased the VEGF-A expression and microvessel density in AF tumor tissues. Furthermore, CLM3 inhibited epidermal growth factor receptor, AKT, and ERK1/2 phosphorylation and down-regulated cyclin D1 in 8305C and AF cells.
  
  
  CONCLUSIONS
  The antitumor and antiangiogenic activity of a pyrazolo[3,4-d]pyrimidine compound (CLM3) is very promising in anaplastic thyroid cancer, opening the way to a future clinical evaluation.
authors.affiliation: Departments of Clinical and Experimental Medicine (A.A., G.B., P.F., S.M.F., C.M., A.F., T.D.D., R.D.), Pharmacy (C.L.M., S.S., F.D.S.), Translational Research and New Technologies in Medicine and Surgery (A.C., S.P.), and Surgical, Medical, Molecular Pathology, and Critical Area (G.M., C.S., G.F., P.M.), University of Pisa, 56126 Pisa, Italy; and Istituto Toscano Tumori (G.B.), 50139 Florence, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Antonelli
authors.forename: Guido
authors.initials: G
authors.lastname: Bocci
authors.forename: Poupak
authors.initials: P
authors.lastname: Fallahi
authors.forename: Concettina
authors.initials: C
authors.lastname: La Motta
authors.forename: Silvia Martina
authors.initials: SM
authors.lastname: Ferrari
authors.forename: Caterina
authors.initials: C
authors.lastname: Mancusi
authors.forename: Anna
authors.initials: A
authors.lastname: Fioravanti
authors.forename: Teresa
authors.initials: T
authors.lastname: Di Desidero
authors.forename: Stefania
authors.initials: S
authors.lastname: Sartini
authors.forename: Alessandro
authors.initials: A
authors.lastname: Corti
authors.forename: Simona
authors.initials: S
authors.lastname: Piaggi
authors.forename: Gabriele
authors.initials: G
authors.lastname: Materazzi
authors.forename: Claudio
authors.initials: C
authors.lastname: Spinelli
authors.forename: Gabriella
authors.initials: G
authors.lastname: Fontanini
authors.forename: Romano
authors.initials: R
authors.lastname: Danesi
authors.forename: Federico
authors.initials: F
authors.lastname: Da Settimo
authors.forename: Paolo
authors.initials: P
authors.lastname: Miccoli
mesh_terms: D020533:Angiogenesis Inhibitors / Q000627:therapeutic use*; D000818:Animals; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009389:Neovascularization, Pathologic / Q000188:drug therapy; D047428:Protein Kinase Inhibitors / Q000627:therapeutic use; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D011720:Pyrazoles / Q000627:therapeutic use*; D011743:Pyrimidines / Q000627:therapeutic use*; D065646:Thyroid Carcinoma, Anaplastic; D013964:Thyroid Neoplasms / Q000098:blood supply / Q000188:drug therapy* / Q000473:pathology; D016896:Treatment Outcome; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
chemical_list: D020533:Angiogenesis Inhibitors; C556683:CLM3 compound; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011743:Pyrimidines; D011505:Protein-Tyrosine Kinases