title: Axitinib and crizotinib combination therapy inhibits bone loss in a mouse model of castration resistant prostate cancer.
abstract: BACKGROUND
  Castration resistant prostate cancer (CRPC) is a leading cause of cancer-related deaths in men. The primary cause of mortality and morbidity in patients is bone metastases and remodeling resulting in osteoblastic and osteolytic lesions. Recently, cabozantinib, a multi-kinase inhibitor (VEGFR2 and c-MET inhibitor), was shown to have efficacy on bone lesions in patients. In this study we tested multi-kinase inhibitors: axitinib (VEGFR inhibitor) and crizotinib (c-MET inhibitor) in a combination trial in mice models.
  
  
  METHODS
  VCaP-Luc cells were grown as subcutaneous implants in intact and castrated NOD-SCID-gamma (NSG) mice to confirm the androgen dependency. For bone metastasis model two cohorts of NSG mice (castrated and intact) received orthotopic injection of VCaP-Luc cells into the bone marrow cavity of left tibia. Mice were monitored weekly for tumor growth using bioluminescence imaging. Animals were randomized into 4 groups based on the tumor bioluminescence signal: vehicle, crizotinib alone, axitinib alone, crizotinib and axitinib in combination. Animals were imaged weekly by in vivo 2-D X-ray imaging to monitor bone remodeling. At the end of the study animals were euthanized and both tibias were extracted for ex vivo high-resolution 3-D micro-computed tomography (μCT) imaging.
  
  
  RESULTS
  Subcutaneous model showed that androgen stimulation may be helpful but not essential for the growth of VCaP-Luc cells. VCaP-Luc cells grown intra-tibially in intact animals caused extensive remodeling of bone with mixed osteoblastic (bone formation) and osteolytic (bone matrix dissolution) lesions. The osteoblastic lesions were predominant and at times extended beyond the tibial shaft into the surrounding tissue. In contrast, only osteolytic lesions were prominent throughout the study in castrated animals. Treatment with crizotinib alone reduced the osteolytic lesions in castrated animals. Axitinib alone reduced the osteoblastic lesions in the intact animals. Combination therapy with axitinib and crizotinib remarkably inhibited the tibial remodeling by VCaP-Luc cells which resulted in a significant reduction of both osteoblastic and osteolytic lesions.
  
  
  CONCLUSIONS
  Our data show that combined inhibition of c-MET and VEGFR can be beneficial for treatment of metastatic bone disease in CRPC and that the drugs act on two different stages of the disease.
authors.affiliation: Global Science and Technology (WCM), Pfizer Global Research and Development, 10724 Science Center Dr, San Diego, CA 92121, USA. jeetendra.eswaraka@pfizer.com.
authors.forename: Jeetendra
authors.initials: J
authors.lastname: Eswaraka
authors.forename: Anand
authors.initials: A
authors.lastname: Giddabasappa
authors.forename: Guangzhou
authors.initials: G
authors.lastname: Han
authors.forename: Kush
authors.initials: K
authors.lastname: Lalwani
authors.forename: Koleen
authors.initials: K
authors.lastname: Eisele
authors.forename: Zheng
authors.initials: Z
authors.lastname: Feng
authors.forename: Timothy
authors.initials: T
authors.lastname: Affolter
authors.forename: James
authors.initials: J
authors.lastname: Christensen
authors.forename: Gang
authors.initials: G
authors.lastname: Li
mesh_terms: D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology / Q000627:therapeutic use*; D000077784:Axitinib; D015519:Bone Density / Q000187:drug effects; D001859:Bone Neoplasms / Q000188:drug therapy* / Q000556:secondary; D045744:Cell Line, Tumor; D000077547:Crizotinib; D006801:Humans; D007093:Imidazoles / Q000008:administration & dosage; D007191:Indazoles / Q000008:administration & dosage; D008297:Male; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D064129:Prostatic Neoplasms, Castration-Resistant / Q000188:drug therapy* / Q000473:pathology; D011720:Pyrazoles / Q000008:administration & dosage; D011725:Pyridines / Q000008:administration & dosage; D023041:Xenograft Model Antitumor Assays
chemical_list: D007093:Imidazoles; D007191:Indazoles; D011720:Pyrazoles; D011725:Pyridines; D000077547:Crizotinib; D000077784:Axitinib