title: Effects of dilution and prolonged storage with preservative in a polyethylene container on Bevacizumab (Avastin™) for topical delivery as a nasal spray in anti-hereditary hemorrhagic telangiectasia and related therapies.
abstract: BACKGROUND
  Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant vascular dysplasia and severe, recurrent epistaxis is a common clinical phenotype associated with HHT. An intranasal treatment regime of diluted Avastin™ (Bevacizumab; recombinant humanized anti-vascular epithelial growth factor immunoglobin G1) using apulsatile nasal irrigator has proven efficacious in clinical practice. However, concerns regarding the stability of Avastin™ following dilution and prolonged storage in standard containers used for drug delivery, such as polyethylene bottles, have so far prevented a more widespread clinical use. Compatibility with the preservative benzalkonium chloride was also unknown.
  
  
  OBJECTIVE
  This study aimed at determining, whether dilution, prolonged refrigerated storage and the presence of the preservative benzalkonium chloride - as required for novel Avastin™ formulations - affected the biochemical and electrochemical properties of the drug.
  
  
  METHODS
  We performed a detailed biochemical and electrochemical analysis of Avastin™, including native and sodium dodecyl sulfate polyacrylamide gel electrophoresis, enzyme-linked immunosorbent assay and isoelectric focusing.
  
  
  RESULTS
  We did not detect any evidence of degeneration or aggregation following dilution and prolonged, refrigerated storage or from the presence of benzalkonium chloride. All biochemical and electrochemical properties of Avastin™ after dilution and prolonged, refrigerated storage were undistinguishable from control.
  
  
  CONCLUSIONS
  Our data provide important insight into the stability of Avastin™ and allow the consideration of novel Avastin™ formulations, including its use in a metered-dose nasal spray for the treatment of HHT and other applications.
authors: Vision Research Center and Department of Ophthalmology, University of Missouri - Kansas City, School of Medicine, Kansas City, MO 64108, USA.
authors: Simon
authors: S
authors: Kaja
authors: Jill D
authors: JD
authors: Hilgenberg
authors: Eric
authors: E
authors: Everett
authors: Scott E
authors: SE
authors: Olitsky
authors: Jim
authors: J
authors: Gossage
authors: Peter
authors: P
authors: Koulen
mesh_terms: D030301:Activin Receptors, Type II / Q000235:genetics / Q000276:immunology*; D000281:Administration, Intranasal; D020533:Angiogenesis Inhibitors / Q000008:administration & dosage / Q000276:immunology* / Q000627:therapeutic use; D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage / Q000276:immunology* / Q000627:therapeutic use; D001548:Benzalkonium Compounds / Q000737:chemistry*; D000068258:Bevacizumab; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004844:Epistaxis / Q000150:complications / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007525:Isoelectric Focusing; D059085:Nasal Sprays; D020959:Polyethylene / Q000737:chemistry; D011310:Preservatives, Pharmaceutical / Q000737:chemistry; D013683:Telangiectasia, Hereditary Hemorrhagic / Q000150:complications / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors* / Q000276:immunology
chemical_list: D020533:Angiogenesis Inhibitors; D061067:Antibodies, Monoclonal, Humanized; D001548:Benzalkonium Compounds; D059085:Nasal Sprays; D011310:Preservatives, Pharmaceutical; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D000068258:Bevacizumab; D020959:Polyethylene; C425364:ACVRL1 protein, human; D030301:Activin Receptors, Type II