title: Novel pyrazolopyrimidine derivatives as tyrosine kinase inhibitors with antitumoral activity in vitro and in vivo in papillary dedifferentiated thyroid cancer.
abstract: OBJECTIVE
  We have studied the antitumoral activity of two new pyrazolo[3,4-d]pyrimidine compounds (CLM3 and CLM29) in primary papillary dedifferentiated thyroid cancer (DePTC) cells.
  
  
  METHODS
  The antiproliferative effect was tested in DePTC cells obtained at reoperation from patients with recurrence of the tumor. The concentrations of CLM3 and CLM29 used in the in vitro experiments were 1, 10, 30, and 50 μm.
  
  
  RESULTS
  Proliferation assays in DePTC cells showed a significant reduction of proliferation by CLM3 and CLM29, which was by 12% with CLM3 (the most potent compound) 10 μm, 43% with CLM3 30 μm, and 60% with CLM3 50 μm. CLM3 and CLM29 increased the percentage of apoptotic cells in DePTC cells dose dependently (P < 0.001) and inhibited migration (P < 0.001). A DePTC cell line (AL) was injected sc in CD nu/nu mice, and tumor masses became detectable 10 d after xenotransplantation. CLM3 (40 mg/kg · die) significantly inhibited tumor growth and weight, and the therapeutic effect was significant starting on the 19th day after cell implantation (4 d after the beginning of treatment). The CLM3-treated group of animals did not show any appreciable toxicity. CLM3 and CLM29 increased thrombospondin-1 expression in the AL cell line. A significant reduction of microvessels and in the percentage of antivascular endothelial growth factor antibody immunoreactivity was observed in the CLM3 treated tumors, with a simultaneous increase of the percentage of necrosis.
  
  
  CONCLUSIONS
  The antitumoral activity of two new pyrazolo[3,4-d]pyrimidine compounds (CLM3, CLM29) in vitro and CLM3 in vivo in DePTC has been shown, opening the way to a future clinical evaluation.
authors.affiliation: Department of Internal Medicine, University of Pisa, School of Medicine, Via Roma 67, I-56100, Pisa, Italy. a.antonelli@med.unipi.it
authors.forename: Alessandro
authors.initials: A
authors.lastname: Antonelli
authors.forename: Guido
authors.initials: G
authors.lastname: Bocci
authors.forename: Concettina
authors.initials: C
authors.lastname: La Motta
authors.forename: Silvia Martina
authors.initials: SM
authors.lastname: Ferrari
authors.forename: Poupak
authors.initials: P
authors.lastname: Fallahi
authors.forename: Anna
authors.initials: A
authors.lastname: Fioravanti
authors.forename: Stefania
authors.initials: S
authors.lastname: Sartini
authors.forename: Michele
authors.initials: M
authors.lastname: Minuto
authors.forename: Simona
authors.initials: S
authors.lastname: Piaggi
authors.forename: Alessandro
authors.initials: A
authors.lastname: Corti
authors.forename: Greta
authors.initials: G
authors.lastname: Alì
authors.forename: Piero
authors.initials: P
authors.lastname: Berti
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: D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D000970:Antineoplastic Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002196:Capillaries / Q000473:pathology; D002291:Carcinoma, Papillary / Q000188:drug therapy* / Q000473:pathology; D002452:Cell Count; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004273:DNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D042282:Microdissection; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D048493:Proto-Oncogene Proteins B-raf / Q000096:biosynthesis / Q000235:genetics; D011720:Pyrazoles / Q000138:chemical synthesis / Q000494:pharmacology*; D011743:Pyrimidines / Q000138:chemical synthesis / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013329:Structure-Activity Relationship; D019699:Thrombospondins / Q000096:biosynthesis / Q000235:genetics; D013964:Thyroid Neoplasms / Q000188:drug therapy* / Q000473:pathology; D023041:Xenograft Model Antitumor Assays
chemical_list: D017304:Annexin A5; D000970:Antineoplastic Agents; C556684:CLM29 compound; C556683:CLM3 compound; D004273:DNA, Neoplasm; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011743:Pyrimidines; D019699:Thrombospondins; C014175:pyrazolo(3,4-d)pyrimidine; D011505:Protein-Tyrosine Kinases; C482120:Braf protein, mouse; D048493:Proto-Oncogene Proteins B-raf