title: Development of a Flow-Through USP-4 Apparatus Drug Release Assay to Evaluate Doxorubicin Liposomes.
abstract: Doxil® is a complex parenteral doxorubicin (DOX) liposome formulation approved by the FDA. For generic doxorubicin liposomes, analyzing the release profile of DOX is important for quality control and comparability studies. However, there is no robust standard drug release assay available for doxorubicin liposomes. In this study, we describe a USP-4 apparatus assay capable of discriminating DOX liposomal formulations based on release profile. Establishment of the assay was hindered by limited DOX release from liposomes in physiological conditions at 37°C. The addition of NH4HCO3to the release media facilitated DOX release proportionally to the salt concentration added but caused precipitation of released drug in USP-4 apparatus. Precipitation of DOX was avoided by adding hydroxypropyl-cyclodextrin (HP-CD) to the release medium. We optimized conditions for DOX release by varying a number of parameters such as: concentration of HP-CD, testing temperature, and concentration of tested samples. The optimized release medium contained: 100 mM NH4HCO3, 75 mM 2-(N-morpholino) ethanesulfonic acid (MES) and 5% w/v HP-CD, 5% w/v sucrose, 0.02% w/v NaN3(pH 6). The drug release assay was performed at 45°C. The optimized release assay can discriminate between DOX liposomal formulations of different compositions, physicochemical properties, and prepared by different manufacturing methods. This indicates that the assay could be used to compare DOX release from generic DOX formulations to the innovator product Doxil®.
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, NCRC, 2800 Plymouth Road, Ann Arbor, Michigan, 48105, USA.|Biointerfaces Institute, University of Michigan, NCRC, 2800 Plymouth Road, Ann Arbor, Michigan, 48105, USA.
authors.forename: Wenmin
authors.initials: W
authors.lastname: Yuan
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, NCRC, 2800 Plymouth Road, Ann Arbor, Michigan, 48105, USA.|Biointerfaces Institute, University of Michigan, NCRC, 2800 Plymouth Road, Ann Arbor, Michigan, 48105, USA.
authors.forename: Rui
authors.initials: R
authors.lastname: Kuai
authors.affiliation: ZoneOne Pharma Inc., UCSF Mission Bay, QB3 Garage, Byers Hall, 1700 4th St., Suite 214, MC2522, San Francisco, California, 94158, USA.
authors.forename: Zhipeng
authors.initials: Z
authors.lastname: Dai
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, NCRC, 2800 Plymouth Road, Ann Arbor, Michigan, 48105, USA.|Biointerfaces Institute, University of Michigan, NCRC, 2800 Plymouth Road, Ann Arbor, Michigan, 48105, USA.|School of Pharmacy, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang, 110016, People's Republic of China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: Office of Generic Drugs, Food and Drug Administration, 10903 New Hampshire Avenue, Silver Spring, Maryland, 20993, USA.
authors.forename: Nan
authors.initials: N
authors.lastname: Zheng
authors.affiliation: Office of Generic Drugs, Food and Drug Administration, 10903 New Hampshire Avenue, Silver Spring, Maryland, 20993, USA.
authors.forename: Wenlei
authors.initials: W
authors.lastname: Jiang
authors.affiliation: ZoneOne Pharma Inc., UCSF Mission Bay, QB3 Garage, Byers Hall, 1700 4th St., Suite 214, MC2522, San Francisco, California, 94158, USA.
authors.forename: Charles
authors.initials: C
authors.lastname: Noble
authors.affiliation: ZoneOne Pharma Inc., UCSF Mission Bay, QB3 Garage, Byers Hall, 1700 4th St., Suite 214, MC2522, San Francisco, California, 94158, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Hayes
authors.affiliation: ZoneOne Pharma Inc., UCSF Mission Bay, QB3 Garage, Byers Hall, 1700 4th St., Suite 214, MC2522, San Francisco, California, 94158, USA.|Department of Bioengineering and Therapeutic Sciences, UCSF, 513 Parnassus Ave., Box 0912, San Francisco, California, 94143, USA.
authors.forename: Francis C
authors.initials: FC
authors.lastname: Szoka
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, NCRC, 2800 Plymouth Road, Ann Arbor, Michigan, 48105, USA. annaschw@umich.edu.|Biointerfaces Institute, University of Michigan, NCRC, 2800 Plymouth Road, Ann Arbor, Michigan, 48105, USA. annaschw@umich.edu.
authors.forename: Anna
authors.initials: A
authors.lastname: Schwendeman
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000737:chemistry* / Q000592:standards; D002626:Chemistry, Pharmaceutical; D002851:Chromatography, High Pressure Liquid; D004317:Doxorubicin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000592:standards; D004339:Drug Compounding; D065546:Drug Liberation; D008081:Liposomes; D010316:Particle Size; D011092:Polyethylene Glycols / Q000737:chemistry / Q000592:standards; D012995:Solubility; D013678:Technology, Pharmaceutical / Q000295:instrumentation* / Q000592:standards
chemical_list: D000903:Antibiotics, Antineoplastic; D008081:Liposomes; C506643:liposomal doxorubicin; D011092:Polyethylene Glycols; D004317:Doxorubicin
keywords: Doxil®; USP-4 apparatus; doxorubicin liposomes; release assay