title: Activity and kinetics of dissociation and transfer of amphotericin B from a novel delivery form.
abstract: Recently it has been demonstrated that moderate heat treatment of Amphotericin B/deoxycholate solutions (HAmB-DOC ) leads to a therapeutically interesting supramolecular rearrangement that can be observed by significant changes in light scattering, CD, and absorbance. In this study, we continue the investigation of the physical properties of this new form by evaluating the activity and kinetics of dissociation and dispersion of HAmB-DOC and AmB-DOC in saline, serum, and in model mammalian or fungal lipid biomimetic membrane vesicles. Stopped-flow spectrophotometry combined with singular value decomposition (SVD) and global analysis were used to resolve the components of this process. The dissociation kinetics for both states are complex, requiring multi-exponential fits, yet in most cases SVD indicates only two significant changing species representing the monomer and the aggregate. The kinetic mechanism could involve dissociation of monomers from coexisting spectroscopically similar but structurally distinct aggregates or sequential rearrangements in supramolecular structure of aggregates. Rate constants and amplitudes of dissociation from aggregates to monomer in buffer, whole serum, 10% cholesterol, and ergosterol membrane vesicles are generally greater for AmB-DOC, demonstrating its greater kinetic instability. In addition, at comparable low concentrations, HAmB-DOC and AmB-DOC are nearly equally active at promoting cation selective permeability in ergosterol-containing membranes; however, HAmB-DOC is much less active against mammalian mimetic cholesterol-containing vesicles, despite a higher level of self-association, supporting previous observations that there exists a specific "toxic aggregate" structure.
authors.affiliation: Department of Chemistry, University of Wisconsin-Eau Claire, Eau Claire, Wisconsin 54702, USA.
authors.forename: B
authors.initials: B
authors.lastname: Baas
authors.forename: K
authors.initials: K
authors.lastname: Kindt
authors.forename: A
authors.initials: A
authors.lastname: Scott
authors.forename: J
authors.initials: J
authors.lastname: Scott
authors.forename: P
authors.initials: P
authors.lastname: Mikulecky
authors.forename: S C
authors.initials: SC
authors.lastname: Hartsel
mesh_terms: D000666:Amphotericin B / Q000097:blood* / Q000737:chemistry* / Q000493:pharmacokinetics; D000818:Animals; D000935:Antifungal Agents / Q000097:blood* / Q000737:chemistry* / Q000493:pharmacokinetics; D002417:Cattle; D018118:Chloride Channels / Q000737:chemistry; D002784:Cholesterol; D003840:Deoxycholic Acid / Q000097:blood* / Q000737:chemistry* / Q000493:pharmacokinetics; D004338:Drug Combinations; D004355:Drug Stability; D004875:Ergosterol; D066298:In Vitro Techniques; D007700:Kinetics; D008567:Membranes, Artificial; D018716:Molecular Mimicry; D010084:Oxidation-Reduction; D015221:Potassium Channels / Q000737:chemistry; D012965:Sodium Chloride; D013050:Spectrometry, Fluorescence
chemical_list: D000935:Antifungal Agents; D018118:Chloride Channels; D004338:Drug Combinations; D008567:Membranes, Artificial; D015221:Potassium Channels; D003840:Deoxycholic Acid; D012965:Sodium Chloride; D000666:Amphotericin B; C059765:amphotericin B, deoxycholate drug combination; D002784:Cholesterol; D004875:Ergosterol