title: Antineoplastic activity of the multitarget tyrosine kinase inhibitors CLM3 and CLM94 in medullary thyroid cancer in vitro.
abstract: BACKGROUND
  We report the antineoplastic and anti-angiogenic activity of the pyrazolo[3,4-d]pyrimidine derivative CLM3 and the cyclic amide CLM94, both multiple tyrosine kinase inhibitors (TKIs), in human primary medullary thyroid cancer (P-MTC) cells, and in vitro in the medullary thyroid cancer (MTC) cell lines TT (harboring a RET C634W activating mutation) and MZ-CRC-1 (carrying the MEN2B RET mutation Met891Thr).
  
  
  METHODS
  The antiproliferative and proapoptotic effects of CLM3 and CLM94 (1, 5, 10, 30, and 50 μmol/L) were tested in P-MTC cells obtained at operation, and in TT cells. In addition, the antiproliferative effects of CLM3 and CLM94 (0.005, 0.05, 0.5, and 5 μmol/L) were tested in TT and MZ-CRC-1 cells after 7 days of treatment to compare the results with those previously reported in the literature.
  
  
  RESULTS
  CLM3 and CLM94 (30 or 50 μmol/L) inhibited (P < .01) the proliferation of the P-MTC cells, TT cells, and MZ-CRC-1 cells and increased the level of apoptosis in a dose-dependent manner at 10, 30, and 50 μmol/L (P < .001), while having no effect on migration or invasion. The inhibition of proliferation by CLM3 and CLM94 was similar among P-MTC cells with/without RET mutations, and similar effects were observed regarding the increased level of apoptosis. Furthermore, CLM3 and CLM94 significantly decreased vascular endothelial growth factor-A expression in TT cells.
  
  
  CONCLUSIONS
  The antitumor activities of the multiple TKIs CLM3 and CLM94 were demonstrated in both primary MTC cultures as well as 2 established MTC cell lines in vitro, opening an avenue for future clinical evaluations.
authors.affiliation: Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: Silvia Martina
authors.initials: SM
authors.lastname: Ferrari
authors.affiliation: Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: Poupak
authors.initials: P
authors.lastname: Fallahi
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Concettina
authors.initials: C
authors.lastname: La Motta
authors.affiliation: Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: Guido
authors.initials: G
authors.lastname: Bocci
authors.affiliation: Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: Alda
authors.initials: A
authors.lastname: Corrado
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Materazzi
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: David
authors.initials: D
authors.lastname: Galleri
authors.affiliation: Department of Translational Research and of New Technologies in Medicine and Surgery, University of Pisa, Pisa, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Piaggi
authors.affiliation: Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: Romano
authors.initials: R
authors.lastname: Danesi
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Federico
authors.initials: F
authors.lastname: Da Settimo
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Miccoli
authors.affiliation: Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy. Electronic address: alessandro.antonelli@med.unipi.it.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Antonelli
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001549:Benzamides / Q000494:pharmacology / Q000627:therapeutic use*; D018276:Carcinoma, Medullary / Q000188:drug therapy*; D018278:Carcinoma, Neuroendocrine; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D011720:Pyrazoles / Q000494:pharmacology / Q000627:therapeutic use*; D011743:Pyrimidines / Q000494:pharmacology / Q000627:therapeutic use*; D012439:Saccharin / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D013964:Thyroid Neoplasms / Q000188:drug therapy*; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors
chemical_list: C572631:4-chloro-N-(1,1,3-trioxo-2,3-dihydrobenzo(d)isothiazol-4-yl)benzamide; D001549:Benzamides; C556683:CLM3 compound; D011720:Pyrazoles; D011743:Pyrimidines; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D011505:Protein-Tyrosine Kinases; D012439:Saccharin