A Scalable Method for Squalenoylation and Assembly of Multifunctional 64 Cu-Labeled Squalenoylated Gemcitabine Nanoparticles.

Autor:	Tucci ST; Department of Biomedical Engineering, University of California Davis, Davis, California, 95616, USA., Seo JW; Department of Radiology, Stanford University, Palo Alto, CA 94304, USA., Kakwere H; Department of Radiology, Stanford University, Palo Alto, CA 94304, USA., Kheirolomoom A; Department of Radiology, Stanford University, Palo Alto, CA 94304, USA., Ingham ES; Department of Biomedical Engineering, University of California Davis, Davis, California, 95616, USA., Mahakian LM; Department of Biomedical Engineering, University of California Davis, Davis, California, 95616, USA., Tam S; Department of Biomedical Engineering, University of California Davis, Davis, California, 95616, USA., Tumbale S; Department of Radiology, Stanford University, Palo Alto, CA 94304, USA., Baikoghli M; Department of Molecular and Cellular Biology, University of California Davis, Davis, California, 95616, USA., Cheng RH; Department of Molecular and Cellular Biology, University of California Davis, Davis, California, 95616, USA., Ferrara KW; Department of Radiology, Stanford University, Palo Alto, CA 94304, USA.
Jazyk:	angličtina
Zdroj:	Nanotheranostics [Nanotheranostics] 2018 Sep 05; Vol. 2 (4), pp. 387-402. Date of Electronic Publication: 2018 Sep 05 (Print Publication: 2018).
DOI:	10.7150/ntno.26969
Abstrakt:	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 (Kras LSL-G12D/+ ; Trp53 LSL-R172H/+ ; Pdx-Cre) pancreatic cancer cell lines. A 64 Cu 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 64 Cu-SqNOTA-SqGem NP. Competing Interests: Competing Interests: The authors have declared that no competing interest exists.
Databáze:	MEDLINE
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