EGFR Targeted Cetuximab-Valine-Citrulline (vc)-Doxorubicin Immunoconjugates- Loaded Bovine Serum Albumin (BSA) Nanoparticles for Colorectal Tumor Therapy.

Abstract

BACKGROUND:

Specific modifications to carriers to achieve targeted delivery of chemotherapeutics into malignant tissues are a critical point for efficient diagnosis and therapy. In this case, bovine serum albumin (BSA) was conjugated with cetuximab-valine-citrulline (vc)-doxorubicin (DOX) to target epidermal growth factor receptor (EGFR) and enable the release of drug in EGFR-overexpressed tumor cells.

METHODS:

Maleimidocaproyl-valine-citrulline-p-aminobenzylcarbonyl-p-nitrophenol (MC-Val-Cit-PAB-PNP) and DOX were used to synthesize MC-Val-Cit-PAB-DOX, which was further linked with cetuximab to prepare antibody-drug conjugates (ADCs). Then, the ADCs were adsorbed to the surface of the BSA nanoparticles (NPs), which were prepared by a desolvation method to obtain cetuximab-vc-DOX-BSA-NPs. The cetuximab-vc-DOX conjugates adsorbed on the surface of the BSA nanoparticles were determined and optimized by size exclusion chromatography. An in vitro cytotoxicity study was conducted using a colon carcinoma cell line with different EGFR-expression levels to test the selectivity of cetuximab-vc-DOX-NPs.

RESULTS:

The vc-DOX and cetuximab-vc-DOX conjugates were both synthesized successfully and their structural characteristics confirmed by 1H-NMR and SDS-PAGE. The MTT assay showed stronger cytotoxicity of cetuximab-vc-DOX-NPs versus control IgG-vc-DOX-NPs in EGFR-overexpressing RKO cells. Cellular binding and intracellular accumulation determined by flow cytometry and confocal laser scanning microscopy revealed the strong binding ability of cetuximab-vc-DOX-NPs to RKO cells. The in vivo imaging study demonstrated that cetuximab-vc-DOX-NPs exhibited higher fluorescent intensity in tumor tissues than non-decorated nanoparticles (IgG-vc-DOX-NPs). In vivo tumor inhibition and survival tests showed that cetuximab-vc-DOX-NPs revealed higher tumor inhibition efficacy and lower systemic toxicity than control IgG-vc-DOX- NPs.

CONCLUSION:

The obtained results emphasize that cetuximab-vc-DOX-NPs, with good tumor-targeting ability and low systemic toxicity, are a promising targeting system for drug delivery.

• Authors: Zixuan Ye, Yue Zhang, Yuanfen Liu, Yanyan Liu, Jiasheng Tu, Yan Shen
• Journal: International journal of nanomedicine (Int J Nanomedicine) Vol. 16 Pg. 2443-2459 ( 2021) ISSN: 1178-2013 [Electronic] New Zealand
• PMID: 33814909 (Publication Type: Journal Article)
• Copyright: © 2021 Ye et al.

Chemical References

• Immunoconjugates
• Serum Albumin, Bovine
• Citrulline
• Doxorubicin
• ErbB Receptors
• Valine
• Cetuximab

Topics

• Adsorption
• Animals
• Cattle
• Cell Line, Tumor
• Cell Survival (drug effects)
• Cetuximab (therapeutic use)
• Citrulline (chemistry)
• Colorectal Neoplasms (drug therapy, pathology)
• Doxorubicin (pharmacology, therapeutic use)
• Drug Delivery Systems (methods)
• Endocytosis (drug effects)
• ErbB Receptors (metabolism)
• Humans
• Immunoconjugates (pharmacology, therapeutic use)
• Mice
• Nanoparticles (chemistry)
• Particle Size
• Proton Magnetic Resonance Spectroscopy
• Serum Albumin, Bovine (chemistry)
• Tissue Distribution (drug effects)
• Valine (chemistry)