Synthesis and in vitro characterization of oxytocin receptor targeted PEGylated immunoliposomes for drug delivery to the uterus

J Liposome Res. 2019 Dec;29(4):357-367. doi: 10.1080/08982104.2018.1556293. Epub 2019 Feb 5.

Abstract

Targeted delivery of therapeutics to the uterus is an important goal in the treatment of obstetric complications, such as preterm labour, postpartum hemorrhage, and dysfunctional labour. Current treatment for these obstetric complications is challenging, as there are limited effective and safe therapeutic options available. We have developed a targeted drug delivery system for the uterus by conjugating anti-oxytocin receptor (OTR) antibodies to the surface of PEGylated liposomes (OTR-PEG-ILs). The functionality of the OTR-PEG-ILs has previously been evaluated on human and murine myometrial tissues as well as in vivo in a murine model of preterm labour. The aim of this study was to report the pharmaceutical synthesis and characterization of the OTR-PEG-ILs and investigate their specific cellular interaction with OTR-expressing myometrial cells in vitro. Immunoliposomes composed of 1,2-distearoyl-sn-glycero-2-phosphocholine (DSPC) and cholesterol were prepared using an optimized method for the coupling of low concentrations of antibody to liposomes. The liposomes were characterized for particle size, antibody conjugation, drug encapsulation, liposome stability, specificity of binding, cellular internalization, mechanistic pathway of cellular uptake, and cellular toxicity. Cellular association studies demonstrated specific binding of OTR-PEG-ILs to OTRs and significant cellular uptake following binding. Evaluation of the mechanistic pathway of cellular uptake indicated that they undergo internalization through both clathrin- and caveolin-mediated mechanisms. Furthermore, cellular toxicity studies have shown no significant effect of OTR-PEG-ILs or the endocytotic inhibitors on cell viability. This study further supports oxytocin receptors as a novel pharmaceutical target for drug delivery to the uterus.

Keywords: Liposomes; nanoparticles; oxytocin receptors; targeted drug delivery; uterus.

MeSH terms

  • Animals
  • Antibodies / chemistry
  • Antibodies / immunology
  • Biological Transport
  • Cell Line
  • Cell Survival / drug effects
  • Cholesterol / chemistry
  • Delayed-Action Preparations / chemistry*
  • Drug Compounding / methods
  • Drug Stability
  • Female
  • Humans
  • Liposomes / chemistry*
  • Mice
  • Myometrium / cytology
  • Nanocapsules / chemistry*
  • Phosphatidylcholines / chemistry
  • Polyethylene Glycols / chemistry
  • Receptors, Oxytocin / immunology*
  • Signal Transduction
  • Surface Properties
  • Uterus / metabolism*

Substances

  • Antibodies
  • Delayed-Action Preparations
  • Liposomes
  • Nanocapsules
  • Phosphatidylcholines
  • Receptors, Oxytocin
  • Polyethylene Glycols
  • Cholesterol