Abstract
Introduction: The majority of nanomedicines and numerous other advanced therapeutics like antibody drugs contain polyethylene glycol (PEG) moieties, intended to improve their pharmacokinetic and pharmacological profiles.
Methods: PEGylated nanomedicines and protein therapeutics, neat and ex vivo, were digested with sulfuric acid to release the PEG moitiies, which were quantidfied by liquid chromatography coupled to mass spectrometry (LC-MS/MS), against matrix-matched standard curves.
Results: Key results obtained include: i) Treatment with strong acid reliably cleaves the PEG entity from the conjugate molecules and also digests tissue effectively, thus providing a simultaneous extraction and normalization of the sample. ii) PEG is well suited for detection by LC-MS/MS with high sensitivity and specificity, depsite the general polydispersity in chain length found in most medical applications. iii) By using LC-MS/MS, it was found that PEG is a suitable marker for a wide range of advanced therapeutics, including liposomes, LNPs, polymeric nanoparticles and therapeutic proteins. iv) The simultaneous hydrolysis of the PEG conjugates and biological tissue matrices enables a robust extraction of PEG after in vivo administration of PEGylated medicines. v) Analysis of PEG biodistribution in tissue with the methodology developed here reveals different drug release kinetics from nanomedicines in vivo. Specifically, the comparison of polymeric nanoparticles containing the antineoplastic agent cabazitaxel and solid lipid nanoparticles containing the near-infrared dye IR780-oleyl for use in fluorescence-guided surgery, was performed. In both cases the small-molecule payload is noncovalently entrapped in the nanoparticles. Quantification of both the PEG moieties and the payloads allowed unprecendented understanding of biodistribution and drug release after admininstration in vivo.
Conclusions/Implications: We consider that the methodlogy developed here is a robust and versatile approach for quantitative analysis of PEGylated therapeutics in biological matrices, and could improve understanding of biodistribution and drug release in vivo. The methodology is similarly applicable to other PEGylated compounds.
Learning Objective 1: To understand how simultanous quantification of PEG moieties from advanced therapeutics and the corresponding bioactive compounds intended for delivery can provide in-depth understanding of biodistribution and pharmacokinetics of complex medicines.
Methods: PEGylated nanomedicines and protein therapeutics, neat and ex vivo, were digested with sulfuric acid to release the PEG moitiies, which were quantidfied by liquid chromatography coupled to mass spectrometry (LC-MS/MS), against matrix-matched standard curves.
Results: Key results obtained include: i) Treatment with strong acid reliably cleaves the PEG entity from the conjugate molecules and also digests tissue effectively, thus providing a simultaneous extraction and normalization of the sample. ii) PEG is well suited for detection by LC-MS/MS with high sensitivity and specificity, depsite the general polydispersity in chain length found in most medical applications. iii) By using LC-MS/MS, it was found that PEG is a suitable marker for a wide range of advanced therapeutics, including liposomes, LNPs, polymeric nanoparticles and therapeutic proteins. iv) The simultaneous hydrolysis of the PEG conjugates and biological tissue matrices enables a robust extraction of PEG after in vivo administration of PEGylated medicines. v) Analysis of PEG biodistribution in tissue with the methodology developed here reveals different drug release kinetics from nanomedicines in vivo. Specifically, the comparison of polymeric nanoparticles containing the antineoplastic agent cabazitaxel and solid lipid nanoparticles containing the near-infrared dye IR780-oleyl for use in fluorescence-guided surgery, was performed. In both cases the small-molecule payload is noncovalently entrapped in the nanoparticles. Quantification of both the PEG moieties and the payloads allowed unprecendented understanding of biodistribution and drug release after admininstration in vivo.
Conclusions/Implications: We consider that the methodlogy developed here is a robust and versatile approach for quantitative analysis of PEGylated therapeutics in biological matrices, and could improve understanding of biodistribution and drug release in vivo. The methodology is similarly applicable to other PEGylated compounds.
Learning Objective 1: To understand how simultanous quantification of PEG moieties from advanced therapeutics and the corresponding bioactive compounds intended for delivery can provide in-depth understanding of biodistribution and pharmacokinetics of complex medicines.