title: Utilizing native fluorescence imaging, modeling and simulation to examine pharmacokinetics and therapeutic regimen of a novel anticancer prodrug.
abstract: Success of cancer prodrugs relying on a foreign gene requires specific delivery of the gene to the cancer, and improvements such as higher level gene transfer and expression. Attaining these objectives will be facilitated in preclinical studies using our newly discovered CNOB-GDEPT, consisting of the produrg: 6-chloro-9-nitro-5-oxo-5H-benzo-(a)-phenoxazine (CNOB) and its activating enzyme ChrR6, which generates the cytotoxic product 9-amino-6-chloro-5H-benzo[a]phenoxazine-5-one (MCHB). MCHB is fluorescent and can be noninvasively imaged in mice, and here we investigated whether MCHB fluorescence quantitatively reflects its concentration, as this would enhance its reporter value in further development of the CNOB-GDEPT therapeutic regimen. PK parameters were estimated and used to predict more effective CNOB administration schedules.
  
  
  
  CNOB (3.3 mg/kg) was injected iv in mice implanted with humanized ChrR6 (HChrR6)-expressing 4T1 tumors. Fluorescence was imaged in live mice using IVIS Spectrum, and quantified by Living Image 3.2 software. MCHB and CNOB were quantified also by LC/MS/MS analysis. We used non-compartmental model to estimate PK parameters. Phoenix WinNonlin software was used for simulations to predict a more effective CNOB dosage regimen.
  
  
  
  CNOB administration significantly prolonged mice survival. MCHB fluorescence quantitatively reflected its exposure levels to the tumor and the plasma, as verified by LC/MS/MS analysis at various time points, including at a low concentration of 2 ng/g tumor. The LC/MS/MS data were used to estimate peak plasma concentrations, exposure (AUC0-24), volume of distribution, clearance and half-life in plasma and the tumor. Simulations suggested that the CNOB-GDEPT can be a successful therapy without large increases in the prodrug dosage.
  
  
  
  MCHB fluorescence quantifies this drug, and CNOB can be effective at relatively low doses. MCHB fluorescence characteristics will expedite further development of CNOB-GDEPT by, for example, facilitating specific gene delivery to the tumor, its prolonged expression, as well as other attributes necessary for successful gene-delivered enzyme prodrug therapy.
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, Sherman Fairchild Science Building, 299 Campus Drive, Stanford, CA, 94305, USA.
authors.forename: Jing-Hung
authors.initials: JH
authors.lastname: Wang
authors.affiliation: Bioanalytical Assays and Pharmacokinetics, Bayer HealthCare LLC, 455 Mission Bay Boulevard South, San Francisco, CA, 94158, USA.
authors.forename: Aaron N
authors.initials: AN
authors.lastname: Endsley
authors.affiliation: Biosciences Division, SRI International, Menlo Park, 94025, CA, USA.
authors.forename: Carol E
authors.initials: CE
authors.lastname: Green
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, Sherman Fairchild Science Building, 299 Campus Drive, Stanford, CA, 94305, USA. a.matin@stanford.edu.
authors.forename: A C
authors.initials: AC
authors.lastname: Matin
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000493:pharmacokinetics*; D045744:Cell Line, Tumor; D004334:Drug Administration Schedule; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D061848:Optical Imaging; D010078:Oxazines / Q000008:administration & dosage / Q000493:pharmacokinetics*; D011355:Prodrugs / Q000008:administration & dosage / Q000493:pharmacokinetics*
chemical_list: C539233:6-chloro-9-nitro-5-oxo-5H-benzo(a)phenoxazine; D000970:Antineoplastic Agents; D010078:Oxazines; D011355:Prodrugs
keywords: CNOB; Cancer; Fluorescence; Imaging; Modeling and simulation; Pharmacokinetics; Prodrug