A scalable method for squalenoylation and assembly of multifunctional64cu-labeled squalenoylated gemcitabine nanoparticles

Samantha T. Tucci, Jai W. Seo, Hamilton Kakwere, Azadeh Kheirolomoom, Elizabeth S. Ingham, Lisa M. Mahakian, Sarah Tam, Spencer Tumbale, Mo Baikoghli, R. Holland Cheng, Katherine W Ferrara

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Squalenoylation of gemcitabine, a front-line therapy for pancreatic cancer, allows for improved cellular-level and system-wide drug delivery. The established methods to conjugate squalene to gemcitabine and to form nanoparticles (NPs) with the squalenoylated gemcitabine (SqGem) conjugate are cumbersome, time-consuming and can be difficult to reliably replicate. Further, the creation of multi-functional SqGem-based NP theranostics would facilitate characterization of in vivo pharmacokinetics and efficacy. Methods: Squalenoylation conjugation chemistry was enhanced to improve reliability and scalability using tert-butyldimethylsilyl (TBDMS) protecting groups. We then optimized a scalable microfluidic mixing platform to produce SqGem-based NPs and evaluated the stability and morphology of select NP formulations using dynamic light scattering (DLS) and transmission electron microscopy (TEM). Cytotoxicity was evaluated in both PANC-1 and KPC (KrasLSL-G12D/+; Trp53LSL-R172H/+; Pdx-Cre) pancreatic cancer cell lines. A 64Cu chelator (2-S-(4-aminobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid, NOTA) was squalenoylated and used with positron emission tomography (PET) imaging to monitor the in vivo fate of SqGem-based NPs. Results: Squalenoylation yields of gemcitabine increased from 15% to 63%. Cholesterol-PEG-2k inclusion was required to form SqGem-based NPs using our technique, and additional cholesterol inclusion increased particle stability at room temperature; after 1 week the PDI of SqGem NPs with cholesterol was ~ 0.2 while the PDI of SqGem NPs lacking cholesterol was ~ 0.5. Similar or superior cytotoxicity was achieved for SqGem-based NPs compared to gemcitabine or Abraxane® when evaluated at a concentration of 10 µM. Squalenoylation of NOTA enabled in vivo monitoring of SqGem-based NP pharmacokinetics and biodistribution. Conclusion: We present a scalable technique for fabricating efficacious squalenoylated-gemcitabine nanoparticles and confirm their pharmacokinetic profile using a novel multifunctional 64Cu-SqNOTA-SqGem NP.

Original languageEnglish (US)
Pages (from-to)387-402
Number of pages16
JournalNanotheranostics
Volume2
Issue number4
DOIs
StatePublished - Jan 1 2018
Externally publishedYes

Keywords

  • Cu
  • Gemcitabine
  • Nanoparticles
  • Pancreatic cancer
  • Radiolabeling
  • Squalene

ASJC Scopus subject areas

  • Biotechnology
  • Medicine (miscellaneous)
  • Biomedical Engineering
  • Pharmacology, Toxicology and Pharmaceutics (miscellaneous)

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  • Cite this

    Tucci, S. T., Seo, J. W., Kakwere, H., Kheirolomoom, A., Ingham, E. S., Mahakian, L. M., Tam, S., Tumbale, S., Baikoghli, M., Cheng, R. H., & Ferrara, K. W. (2018). A scalable method for squalenoylation and assembly of multifunctional64cu-labeled squalenoylated gemcitabine nanoparticles. Nanotheranostics, 2(4), 387-402. https://doi.org/10.7150/ntno.26969