title: A novel alkylating agent, glufosfamide, enhances the activity of gemcitabine in vitro and in vivo.
abstract: Glufosfamide is an alkylating agent consisting of iphosphoramide mustard conjugated to glucose that is currently included in clinical studies of pancreatic cancer. We studied the effects of glufosfamide, in combination with gemcitabine, on in vitro and in vivo models of pancreatic cancer. In proliferation assays, glufosfamide and gemcitabine inhibited the growth of MiaPaCa-2, H766t, and PANC-1 cells, but the combination of the two agents provided greater effects. Apoptosis of MiaPaCa-2 cells, measured by fluorescence-activated cell sorting, was enhanced by the combination of the two drugs, compared to single-agent treatment. Glufosfamide alone inhibited the growth of red fluorescent protein-expressing MiaPaCa-2 tumors in an orthotopic nude mouse model in a dose-dependent manner. Combining glufosfamide (30 mg/kg) with gemcitabine resulted in enhanced inhibition of tumor growth and significantly prolonged survival. Immunohistochemistry of excised tumors revealed that both glufosfamide and gemcitabine increased levels of apoptosis (measured by terminal deoxynucleotidyl transferase-mediated nick end labeling staining) and reduced proliferation (measured by proliferating cell nuclear antigen staining). No effects on microvessel density were observed. These results support the use of the alkylating agent glufosfamide and the DNA synthesis inhibitor gemcitabine, rather than the use of either agent alone, to provide greater benefits and demonstrate that this combination treatment should be useful in the clinical treatment of pancreatic carcinoma.
authors.affiliation: Department of Pharmacology and Toxicology, Threshold Pharmaceuticals, Inc., Redwood City, CA 94061, USA. wsammons@thresholdpharm.com
authors.forename: W Steve
authors.initials: WS
authors.lastname: Ammons
authors.forename: Jin-Wei
authors.initials: JW
authors.lastname: Wang
authors.forename: Zhijian
authors.initials: Z
authors.lastname: Yang
authors.forename: George F
authors.initials: GF
authors.lastname: Tidmarsh
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hoffman
mesh_terms: D000477:Alkylating Agents / Q000494:pharmacology* / Q000627:therapeutic use; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D004357:Drug Synergism; D005947:Glucose / Q000031:analogs & derivatives; D007069:Ifosfamide / Q000031:analogs & derivatives; D051379:Mice; D008819:Mice, Nude; D010190:Pancreatic Neoplasms / Q000188:drug therapy / Q000473:pathology; D010752:Phosphoramide Mustards / Q000494:pharmacology* / Q000627:therapeutic use; D023041:Xenograft Model Antitumor Assays / Q000379:methods; D000093542:Gemcitabine