A novel nucleolin-binding peptide for Cancer Theranostics

Abstract

Background: Cancer-specific ligands have been of great interest as pharmaceutical carriers due to the potential for site-specific delivery. In particular, cancer-specific peptides have many advantages over nanoparticles and antibodies, including high biocompatibility, low immunogenicity, and the formation of nontoxic metabolites. The goal of the present study was the development of a novel cancer-specific ligand. Methods: Cancer-specific peptide ligands were screened using a one-bead-one-compound (OBOC) combinatorial method combined with a multiple-antigen-peptide (MAP) synthesis method. The specificity of the peptide ligands toward cancer cells was tested in vitro using a whole-cell binding assay, flow cytometry, and fluorescence confocal microscopy. The tissue distribution profile and therapeutic efficacy of a paclitaxel (PTX)-conjugated peptide ligand was assessed in vivo using xenograft mouse models. Results: We discovered that AGM-330 specifically bound to cancer cells in vitro and in vivo . Treatment with PTX-conjugated AGM-330 dramatically inhibited cancer cell growth in vitro and in vivo compared to treatment with PTX alone. The results of pull-down assay and LC-MS/MS analyses showed that membrane nucleolin (NCL) was the target protein of AGM-330. Although NCL is known as a nuclear protein, we observed that it was overexpressed on the membranes of cancer cells. In particular, membrane NCL neutralization inhibited growth in cancer cells in vitro . Conclusions: In summary, our findings indicated that NCL-targeting AGM-330 has great potential for use in cancer diagnosis and targeted drug delivery in cancer therapy.

Synthesis of Poly(Malic Acid) Derivatives End-Functionalized with Peptides and Preparation of Biocompatible Nanoparticles to Target Hepatoma Cells

“…Ever since, tremendous research work has been done for designing efficient site-specific drugs using various ligand molecules, such as monoclonal antibodies (mAbs) [ 2 , 3 , 4 ], vitamins [ 2 , 5 , 6 , 7 ], carbohydrates [ 2 , 8 ], and aptamers [ 9 , 10 , 11 ]. More recently, short synthetic peptides have gained interest as targeting agents in the design of site-specific nanomedicines for therapy and/or diagnosis of several pathologies, especially cancers [ 2 , 12 , 13 , 14 , 15 , 16 ].…”

“…Twenty years later (2018), Lutathera ® , which consists of octreotate, another somatostatin analog, grafted to 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) chelating the lutetium 177 ( 177 Lu), was the first FDA-approved radiopharmaceutical for the therapy of SSTR2-positive NETs [ 12 , 28 ]. Since the emergence of somatostatin and its analogs, numerous peptides have been identified by different technologies, such as phage-display, for their high degree of specificity towards cancer cells [ 12 , 13 , 14 , 15 , 16 ]. Such peptides, which are usually easily synthesized on large scales at a low cost, can be chemically modified to be grafted on various materials, such as lipids or polymers, to engineer nanocarriers characterized by their good biocompatibility in addition to their excellent targeting properties [ 12 , 13 , 14 , 15 , 16 , 17 ].…”

Novel role of circRSU1 in the progression of osteoarthritis by adjusting oxidative stress

“…All images were quantified using Image-Pro Plus 6.0 (NIH, Bethesda, MD, USA). Log 2 (fold of change) was used to calculate the relative gray value of specific proteins normalized to β-actin or GAPDH, for western blot results, positive cell percentage for IHC analysis 49 , 50 , and relative fluorescence intensity for immunofluorescence analysis 51 as previously indicated. Enzyme-linked immunosorbent assay (ELISA) was used to detect and quantify pro-inflammatory factors (IL-1β, IL-6, and TNF-a) in the culture supernatant, using a kit from KeyGen (Nanjing, China), according to the manufacturer's instructions.…”

“…A fluorescence in situ hybridization (FISH) kit (RiboBio, Guangzhou, China) was used to detect the probe signals in primary chondrocytes and tissue sections according to the manufacturer's instructions. Images were acquired using a Nikon A1Si Laser Scanning Confocal Microscope (Nikon Instruments Inc., Japan), processed with Image-Pro Plus 6.0 (NIH, Bethesda, MD, USA), and reported as the relative fluorescence intensity as previously indicated 51 .…”

Cancer Stem Cells and Nucleolin as Drivers of Carcinogenesis

“…More recently, AGM-330 peptide was demonstrated to specifically bind nucleolin on the surface of cancer cells in vivo and in vitro. The conjugation of paclitaxel with AGN-330 improved cancer cell growth compared to treatment with paclitaxel alone [ 187 ].…”