Amphiphilic Poly(N-vinylpyrrolidone) Nanoparticles Conjugated with DR5-Specific Antitumor Cytokine DR5-B for Targeted Delivery to Cancer Cells

Abstract

Nanoparticles based on the biocompatible amphiphilic poly(N-vinylpyrrolidone) (Amph-PVP) derivatives are promising for drug delivery. Amph-PVPs self-aggregate in aqueous solutions with the formation of micellar nanoscaled structures. Amph-PVP nanoparticles are able to immobilize therapeutic molecules under mild conditions. As is well known, many efforts have been made to exploit the DR5-dependent apoptosis induction for cancer treatment. The aim of the study was to fabricate Amph-PVP-based nanoparticles covalently conjugated with antitumor DR5-specific TRAIL (Tumor necrosis factor-related apoptosis-inducing ligand) variant DR5-B and to evaluate their in vitro cytotoxicity in 3D tumor spheroids. The Amph-PVP nanoparticles were obtained from a 1:1 mixture of unmodified and maleimide-modified polymeric chains, while DR5-B protein was modified by cysteine residue at the N-end for covalent conjugation with Amph-PVP. The nanoparticles were found to enhance cytotoxicity effects compared to those of free DR5-B in both 2D (monolayer culture) and 3D (tumor spheroids) in vitro models. The cytotoxicity of the nanoparticles was investigated in human cell lines, namely breast adenocarcinoma MCF-7 and colorectal carcinomas HCT116 and HT29. Notably, DR5-B conjugation with Amph-PVP nanoparticles sensitized resistant multicellular tumor spheroids from MCF-7 and HT29 cells. Taking into account the nanoparticles loading ability with a wide range of low-molecular-weight antitumor chemotherapeutics into hydrophobic core and feasibility of conjugation with hydrophilic therapeutic molecules by click chemistry, we suggest further development to obtain a versatile system for targeted drug delivery into tumor cells.

DR5-Selective TRAIL Variant DR5-B Functionalized with Tumor-Penetrating iRGD Peptide for Enhanced Antitumor Activity against Glioblastoma

“…To obtain fluorescently labeled protein DR5-B-iRGD, first the DR5-B-iRGD amino acid sequence was genetically modified at the N-end by replacing the amino acid residue valine in the 114 position to cysteine by site-directed mutagenesis. The cysteine-modified proteins DR5-B and DR5-B-iRGD were obtained by the method earlier reported for DR5-B [ 45 ]. Further, the cysteine-modified proteins DR5-B and DR5-B-iRGD were labeled by maleimide chemistry coupling with the fluorescent dye sulfo-Cyanine 3 maleimide (Lumiprobe, Moscow, Russia) according to the manufacturer’s protocol.…”

Amphiphilic Poly-N-vinylpyrrolidone Nanoparticles as Carriers for Nonsteroidal, Anti-Inflammatory Drugs: Pharmacokinetic, Anti-Inflammatory, and Ulcerogenic Activity Study

“…In our previous studies [ 5 , 6 , 12 , 26 ], we determined the IMC loading efficiency, capacity, and stability of Amph-PVP derivatives with a different molecular weight of polymer hydrophilic fragment and length of anchor long-chain aliphatic hydrophobic fragment. Furthermore, we assessed the in vitro IMC release performance and the cytotoxicity of these nanoparticles [ 11 , 12 , 26 ]. The respective studies demonstrated the excellent biocompatibility and performance of PVP nanoparticles, duplicated in an in vivo acute toxicity determination model.…”

“…According to the method previously developed by our group, Amph-PVP polymer (PVP-OD4000) was synthesized by free-radical polymerization of N-vinylpyrrolidone monomer in the presence of azobisisobutyronitrile (AIBN) as initiator and mercaptoacetic acid (MAA) as chain-growth regulator. The following coupling reaction between carboxyl end-capped poly- N -vinylpyrrolidone with aliphatic n-octadecylamine (OD) in the presence of N, N’-dicyclohexylcarbodiimide (DCC) resulted in the desired polymer [ 24 , 25 , 26 ]. The resulting PVP-OD4000 amphiphilic polymer consists of a hydrophilic macromolecular fragment of PVP with 4000 Da molecular weight and one anchor hydrophobic n-alkyl octadecyl group.…”

Recent Advances in the Development of Nanodelivery Systems Targeting the TRAIL Death Receptor Pathway

“…The obtained P-DR5-B nanoparticles enhanced the cytotoxicity of free DR5-B proteins in vitro in monolayer cell cultures and multicellular tumor spheroids of colon carcinomas HCT116 and HT29 and breast adenocarcinoma MCF-7 cells without cytotoxicity to normal cells. Taking into account the Amph-PVP ability to be loaded with a wide range of low-molecular-weight antitumor chemotherapeutics into hydrophobic core and the feasibility of conjugation with hydrophilic therapeutic molecules, it can be further developed into a versatile system for targeted drug delivery to tumor cells [ 98 ].…”