Trastuzumab- and Fab' fragment-modified curcumin PEG-PLGA nanoparticles: preparation and evaluation in vitro and in vivo.
Introduction
Nanoparticles (NPs) modified with bio-ligands represent a promising strategy for active targeted drug delivery to tumour. However, many targeted ligands, such as trastuzumab (TMAB), have high molecular weight, limiting their application for targeting. In this study, we prepared Fab' (antigen-binding fragments cut from TMAB)-modified NPs (Fab'-NPs) with curcumin (Cur) as a model drug for more effective targeting of human epidermal growth factor receptor 2 (HER2/ErbB2/Neu), which is overexpressed on breast cancer cells.


Material and methods
The release kinetics was conducted by dialysis bags. The ability to kill HER2-overexpressing BT-474 cells of Fab'-Cur-NPs compared with TMAB-Cur-NPs was conducted by cytotoxicity experiments. Qualitative and quantitative cell uptake studies using coumarin-6 (fluorescent probe)-loaded NPs were performed by fluorescence microscopy and flow cytometry. Pharmacokinetics and biodistribution experiments in vivo were assessed by liquid chromatography-tandem mass spectrometry (LC-MS/MS).


Results
The release kinetics showed that both Fab'-Cur-NPs and TMAB-Cur-NPs provided continuous, slow release of curcumin for 72 h, with no significant difference. In vitro cytotoxicity experiments showed that Fab'-Cur-NPs manifested prominent ability to kill HER2-overexpressing BT-474 cells compared with TMAB-Cur-NPs. Qualitative and quantitative cell uptake studies indicated that the accumulation of Fab'-NPs was greater than that of TMAB-NPs in BT-474 (HER2+) cells; However, there was no significant difference in MDA-MB-231 (HER2-) cells. Pharmacokinetics and biodistribution experiments in vivo demonstrated that the half-life (t1/2) and area under the blood concentration-time curve (AUC0-t) of Fab'-Cur-NPs increased 5.30-fold and 1.76-fold relative to those of TMAB-Cur-NPs, respectively. Furthermore, the tumor accumulation of Fab'-Cur-NPs was higher than that of TMAB-Cur-NPs.


Conclusion
Fab' fragment has greater capacity than the intact antibody to achieve tumor targeting through NP-based delivery.
['Duan|Dongyu|D|', 'Wang|Aiping|A|', 'Ni|Ling|L|', 'Zhang|Liping|L|', 'Yan|Xiuju|X|', 'Jiang|Ying|Y|', 'Mu|Hongjie|H|', 'Wu|Zimei|Z|', 'Sun|Kaoxiang|K|', 'Li|Youxin|Y|']
D000818:Animals D016923:Cell Death D045744:Cell Line, Tumor D003374:Coumarins D003474:Curcumin D004705:Endocytosis D005260:Female D005434:Flow Cytometry D006801:Humans D007140:Immunoglobulin Fab Fragments D007275:Injections, Intravenous D008807:Mice, Inbred BALB C D008856:Microscopy, Fluorescence D053758:Nanoparticles D011091:Polyesters D011092:Polyethylene Glycols D017207:Rats, Sprague-Dawley D013844:Thiazoles D014018:Tissue Distribution D000068878:Trastuzumab
breast cancer pharmacokinetics biodistribution Fab′-modified curcumin nanoparticles trastuzumab-modified curcumin nanoparticles tumour targeting