Preparation and Characterization of Nanoliposomal Irinotecan

Nanoliposome (Nal) and nanoliposomal irinotecan (Nal–IRI) were prepared following freeze–thaw extrusion method [21 (link), 22 (link), 48 (link), 49 (link)]. Briefly, cholesterol, dipalmitoylphosphatidylcholine (DPPC), distearoyl-phosphatidylethanolamine-methoxy polyethylene glycol (DSPE-mPEG2000), distearoyl-glycerophosphoethanolamine-N-dibenzocyclooctyl polyethylene glycol (DSPE-mPEG2000-DBCO), and dioleoylglycerophosphoglycerol (DOPG; Avanti) were mixed at a molar ratio of 2.8:6:0.4:0.2:0.6. For selectivity and uptake studies, 0.1 mol% of dipalmitoylglycero-phosphoethanolamine-N-(lissamine Rhodamine B sulfonyl) (16:0 Liss Rhod PE) was added to the lipid film. The dried lipid film was hydrated with deionized water with or without irinotecan (3 mM) prior to freeze–thaw cycling (4°C –45 °C). Multi-laminar nanoliposomes were then extruded through polycarbonate membrane (Whatman; 0.1 μm) at 45 °C and dialyzed against PBS to remove free irinotecan. Zetasizer NanoZS (Malvern) determined the size and zeta potential of Nals. The concentration of irinotecan was determined using UV–Vis spectroscopy and the established molar extinction coefficients (Additional file 1: Table S3) [21 (link), 22 (link), 48 (link)].

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Liang B.J., Pigula M., Baglo Y., Najafali D., Hasan T, & Huang H.C. (2020). Breaking the selectivity-uptake trade-off of photoimmunoconjugates with nanoliposomal irinotecan for synergistic multi-tier cancer targeting. Journal of Nanobiotechnology, 18, 1.

Publication 2020

polycarbonate membrane NanoZS

Cholesterol Dipalmitoylphosphatidylcholine Dopg Dspe Extinction Freeze Glycerophosphoethanolamine Irinotecan Lipid Lissamine rhodamine b Membrane Methoxy polyethylene glycol Molar Phosphatidylethanolamine Phosphoethanolamine Polycarbonate Polyethylene glycol Selectivity Spectroscopy

Corresponding Organization :

Other organizations : University of Maryland, College Park, Massachusetts General Hospital, Harvard University

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Variable analysis

independent variables

Preparation method: freeze-thaw extrusion

dependent variables

Size of nanoliposomes (Nals)
Zeta potential of Nals
Concentration of irinotecan in Nal-IRI

control variables

Lipid composition: cholesterol, dipalmitoylphosphatidylcholine (DPPC), distearoyl-phosphatidylethanolamine-methoxy polyethylene glycol (DSPE-mPEG2000), distearoyl-glycerophosphoethanolamine-N-dibenzocyclooctyl polyethylene glycol (DSPE-mPEG2000-DBCO), and dioleoylglycerophosphoglycerol (DOPG)
Lipid molar ratio: 2.8:6:0.4:0.2:0.6
Inclusion of 0.1 mol% dipalmitoylglycero-phosphoethanolamine-N-(lissamine Rhodamine B sulfonyl) (16:0 Liss Rhod PE) for selectivity and uptake studies
Hydration of dried lipid film with deionized water with or without irinotecan (3 mM)

controls

Positive control: Not specified
Negative control: Not specified

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