Physicochemical surrogates for in vitro toxicity assessment of liposomal amphotericin B

Richard Marx; Jaeweon Lee; Yuri Svirkin; Seongkyu Yoon; Nelson Landrau; Md Abul Kaisar; Bin Qin; Jin H Park; Khondoker Alam; Darby Kozak; Yan Wang; Xiaoming Xu; Jiwen Zheng; Benjamin Rivnay

doi:10.1016/j.ijpharm.2022.122273

Physicochemical surrogates for in vitro toxicity assessment of liposomal amphotericin B

Int J Pharm. 2022 Nov 25:628:122273. doi: 10.1016/j.ijpharm.2022.122273. Epub 2022 Oct 11.

Authors

Affiliations

¹ Landrau Scientific Innovations, 22 Laurel Street, Leominster, MA 01453, USA.
² Department of Chemical Engineering, University of Massachusetts Lowell, 1 University Ave, Lowell, MA, 01854, USA. Electronic address: Seongkyu_Yoon@uml.edu.
³ Food and Drug Administration, Center for Drug Evaluation and Research, Office of Generic Drugs, Office of Research and Standards, 10903 New Hampshire Avenue, Silver Spring, MD 20993, USA.
⁴ Food and Drug Administration, Center for Drug Evaluation and Research, Office of Pharmaceutical Quality, Office of Testing and Research, 10903 New Hampshire Avenue, Silver Spring, MD 20993, USA.
⁵ Food and Drug Administration, Center for Devices and Radiological Health, Office of Science and Engineering Laboratories, 10903 New Hampshire Avenue, Silver Spring, MD 20993, USA.

PMID: 36228881
DOI: 10.1016/j.ijpharm.2022.122273

Abstract

Pharmaceutical toxicity evaluations often use in vitro systems involving primary cells, cell lines or red blood cells (RBCs). Cell-based analyses ('bioassays') can be cumbersome and typically rely on hard-to-standardize biological materials. Amphotericin B (AmB) toxicity evaluations are primarily based on potassium release from RBCs and share these limitations. This study evaluates the potential substitution of two physicochemical AmB toxicity approaches for the bioassay: Ultraviolet-visible spectroscopy (UV-vis) and in vitro drug release kinetics. UV-vis spectral analyses indicated that liposomal AmB's (L-AmB) main peak position (λ_max) and peak ratio (OD₃₄₆/OD₃₂₂) are potential toxicity surrogates. Similarly, two first-order release parameters derived from USP-4 in vitro drug release analyses also provided linear relationships with toxicity. These were the initial, overall drug release rate and the ratio of loose to tight AmB pools. Positive slopes and high correlation coefficients (R² > 0.9) characterized all interrelations between physicochemical parameters and toxicity. These tests converted the manufacturing variables' nonlinear (i.e., curvilinear) relationships with in vitro toxicity to linear responses. Three different toxicity attenuation approaches (2 manufacturing, 1 formulation), covering formulation composition and process aspects, support this approach's universality. These data suggest that one or more spectral and kinetic physicochemical tests can be surrogates for L-AmB in vitro toxicity testing.

Keywords: Aggregation; Amphotericin B; Drug release kinetics; In vitro toxicity; Liposomes; Surrogate test; UV–vis spectra.

MeSH terms

Amphotericin B* / chemistry
Amphotericin B* / toxicity
Antifungal Agents* / chemistry
Antifungal Agents* / toxicity
Drug Liberation
Liposomes

Substances

liposomal amphotericin B
Amphotericin B
Antifungal Agents
Liposomes