Preparation of liposomes for placental targeting and UPLC quantification in tissue of treated mice
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Date
2023-06-26
Authors
Sánchez Téllez, Alma Athenas
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Abstract
In previous work, neutral DOPE-NBD (Ex: 465 nm Em: 535 nm) labelled liposomes were prepared to investigate their internalization in in vitro and ex vivo models. For next experiments in vivo, the labelling probe must be changed to escape from the wavelengths range of autofluorescence present in biological tissue. The far-red fluorescent probe Cyanine 5 (Cy5) linked to the phospholipid DOPE (DOPE-Cy5) (Ex: 640 nm Em: 665 nm) seems a promising alternative to incorporate and label liposomes for this project.
The objectives of this work are to prepare formulations injected to mice and to develop tools for their evaluation. Neutral liposomes which are fluorescently labelled thanks to incorporation of DOPE-Cy5 and composed of various proportion of PEG are decorated with a VAR2CSA peptide and characterized. For coupling with live animal imagery and to assay biodistribution, a quantification method must be developed to quantify DOPE-Cy5 present in different organs of treated mice.
The liposomal formulations’ functionalization capacity was not decremented with the inclusion of new fluorescent phospholipid. The DDS physicochemical characteristics remained apparently the same but further measurements are required. An UPLC method for DOPE-Cy5 quantification in biological tissue was successfully developed and implemented in liver extractions of treated mice with different liposomal formulations. The mobile phase consisted of a mix of methanol, isopropanol, and water. The retention time for DOPE Cy5 resulted at 7.6 min. The treated liver showed concentrations up to 4 μg/L of tissue of DOPE-Cy5 after 24 hours.
As far as we are aware this is the first-time fluorescent lipid from DDS has been detected by UPLC in a in vivo model. Showing the potential application of the method for pharmacokinetic analysis.
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Keywords
Placental diseases, Pregnancy