CLM29, a multi-target pyrazolopyrimidine derivative, has anti-neoplastic activity in medullary thyroid cancer in vitro and in vivo

Alessandro Antonelli; Guido Bocci; Concettina La Motta; Silvia Martina Ferrari; Poupak Fallahi; Alda Corrado; Anna Fioravanti; Stefania Sartini; Paola Orlandi; Simona Piaggi; Alessandro Corti; Gabriele Materazzi; David Galleri; Salvatore Ulisse; Gabriella Fontanini; Romano Danesi; Federico Da Settimo; Paolo Miccoli

doi:10.1016/j.mce.2014.06.002

CLM29, a multi-target pyrazolopyrimidine derivative, has anti-neoplastic activity in medullary thyroid cancer in vitro and in vivo

Mol Cell Endocrinol. 2014 Aug 5;393(1-2):56-64. doi: 10.1016/j.mce.2014.06.002. Epub 2014 Jun 12.

Authors

Alessandro Antonelli¹, Guido Bocci², Concettina La Motta³, Silvia Martina Ferrari⁴, Poupak Fallahi⁵, Alda Corrado⁶, Anna Fioravanti⁷, Stefania Sartini⁸, Paola Orlandi⁹, Simona Piaggi¹⁰, Alessandro Corti¹¹, Gabriele Materazzi¹², David Galleri¹³, Salvatore Ulisse¹⁴, Gabriella Fontanini¹⁵, Romano Danesi¹⁶, Federico Da Settimo¹⁷, Paolo Miccoli¹⁸

Affiliations

¹ Department of Clinical and Experimental Medicine, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: alessandro.antonelli@med.unipi.it.
² Department of Clinical and Experimental Medicine, University of Pisa, Via Savi 10, 56126 Pisa, Italy; Istituto Toscano Tumori, Via Alderotti 27/n, Florence, Italy. Electronic address: guido.bocci@med.unipi.it.
³ Department of Pharmacy, University of Pisa, Via Bonanno Pisano 6, 56126 Pisa, Italy. Electronic address: lamotta@farm.unipi.it.
⁴ Department of Clinical and Experimental Medicine, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: sm.ferrari@int.med.unipi.it.
⁵ Department of Clinical and Experimental Medicine, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: puopak@int.med.unipi.it.
⁶ Department of Clinical and Experimental Medicine, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: corradoalda@gmail.com.
⁷ Department of Clinical and Experimental Medicine, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: anna.fioravanti@libero.it.
⁸ Department of Pharmacy, University of Pisa, Via Bonanno Pisano 6, 56126 Pisa, Italy. Electronic address: stefania.sartini@for.unipi.it.
⁹ Department of Clinical and Experimental Medicine, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: paolaorlandi21@libero.it.
¹⁰ Department of Translational Research and of New Technologies in Medicine and Surgery, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: simona.piaggi@dps.unipi.it.
¹¹ Department of Translational Research and of New Technologies in Medicine and Surgery, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: alessandro.corti@med.unipi.it.
¹² Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: g.materazzi@med.unipi.it.
¹³ Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: galleridavid@hotmail.com.
¹⁴ Department of Experimental Medicine, "Sapienza" University of Rome, Viale dell'Università 30, 00185 Rome, Italy. Electronic address: salvatore.ulisse@uniroma1.it.
¹⁵ Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: gabriella.fontanini@med.unipi.it.
¹⁶ Department of Clinical and Experimental Medicine, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: romano.danesi@med.unipi.it.
¹⁷ Department of Pharmacy, University of Pisa, Via Bonanno Pisano 6, 56126 Pisa, Italy. Electronic address: federico.dasettimo@farm.unipi.it.
¹⁸ Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Via Savi 10, 56126 Pisa, Italy. Electronic address: paolo.miccoli@dc.unipi.it.

PMID: 24931161
DOI: 10.1016/j.mce.2014.06.002

Abstract

CLM29 (a pyrazolo[3,4-d]pyrimidine, that inhibits RET, epidermal growth factor receptor, vascular endothelial growth factor receptor, and has an anti-angiogenic activity) has anti-neoplastic activity in papillary dedifferentiated thyroid cancer. Here we tested CLM29 in medullary thyroid cancer (MTC), in primary MTC cells (P-MTC) obtained at surgery, and in TT cells harboring (C634W) RET mutation. CLM29 (10, 30, 50 μM) inhibited significantly (P<0.001) the proliferation, and increased the percentage of apoptotic P-MTC, TT and human dermal microvascular endothelial cells. The inhibition of proliferation by CLM29 was similar in P-MTC cells with/without RET mutation. TT cells were injected sc in CD nu/nu mice, and tumor masses became detectable between 20 and 30 days after xenotransplantation; CLM29 (50mg/kg/die) reduced significantly tumor growth and weight, and microvessel density. The anti-tumor activity of CLM29 has been shown in MTC in vitro, and in vivo, opening the way to a future clinical evaluation.

Keywords: CLM29; Medullary thyroid cancer; Pyrazolo[3,4-d]pyrimidine; Thyroid cancer; Tyrosine kinase inhibitors.

MeSH terms

Animals
Antineoplastic Agents / pharmacology
Antineoplastic Agents / therapeutic use
Carcinoma, Neuroendocrine
Cell Proliferation / drug effects
Humans
Mice
Mice, Nude
Pyrazoles / pharmacology*
Pyrazoles / therapeutic use*
Pyrimidines / pharmacology*
Pyrimidines / therapeutic use*
Real-Time Polymerase Chain Reaction
Thyroid Neoplasms / drug therapy*
Tumor Cells, Cultured
Vascular Endothelial Growth Factor A / metabolism

Substances

Antineoplastic Agents
CLM29 compound
Pyrazoles
Pyrimidines
Vascular Endothelial Growth Factor A

Supplementary concepts

Thyroid cancer, medullary