Celastrol Self-Stabilized Nanoparticles for Effective Treatment of Melanoma.

BACKGROUND: Celastrol (CEL), a triterpene extracted from the Chinese herb tripterygium wilfordii, has been reported to have profound anticancer activities. However, poor water solubility and high side toxicities have severely restricted the clinical applications of CEL. PURPOSE: We proposed a facile "in situ drug conjugation-induced self-assembly" strategy to prepare CEL-loaded nanoparticles (CEL-NPs) that exhibited enhanced antitumor activity against melanoma. METHODS: First, the CEL was chemically conjugated onto a methoxyl poly(ethylene glycol)-b-poly(L-lysine) (mPEG-PLL) backbone, resulting in the conversion of the double hydrophilic mPEG-PLL polymer into an amphiphilic polymer prodrug, mPEG-PLL/CEL. The obtained mPEG-PLL/CEL could self-assemble into stable micelles in aqueous solution due to the hydrophobic association of CEL moieties in the side chains and the possible electrostatic interaction between the carboxyl group in CEL and the residue amine group in the PLL segment. Thus, the obtained mPEG-PLL/CEL nanoparticles were named CEL self-stabilized nanoparticles (CEL-NPs), which were then characterized by dynamic light scattering and transmission electron microscopy. Furthermore, the antitumor effects of the CEL-NPs were investigated by an MTT assay in vitro and in a B16F10 tumor-bearing mice model. RESULTS: The CEL-NPs exhibited sustained drug release behavior and were effectively endocytosed by B16F10 cells. Furthermore, the in vivo antitumor evaluation demonstrated that the CEL-NPs had remarkably higher tumor growth inhibition rates and lower systemic side effects than free CEL. CONCLUSION: In summary, our present work not only demonstrates the generation of stable CEL-loaded nanoparticles for the efficient treatment of melanoma but also describes a general way to prepare drug self-stabilized nanomedicine for anticancer therapy.

Hypoxia-Responsive Polypeptide Nanoparticles Loaded with Doxorubicin for Breast Cancer Therapy.

Microenvironments of various solid tumors are characterized by hypoxia. Herein, we report a novel nanoparticle that can selectively release loaded drugs in hypoxic environments. The nanoparticle was prepared using a hypoxia-responsive amphiphilic polymer in aqueous media. The polymer was synthesized by conjugating a hydrophobic small molecule, 4-nitrobenzyl (3-azidopropyl) carbamate, to the side chains of an mPEG-PPLG copolymer. Doxorubicin (DOX) could be loaded into the nanoparticles with a high efficiency of 97.8%. The generated drug-loaded micellar nanoparticles (PPGN@DOX) presented hypoxia-sensitive drug release behavior in vitro. Meanwhile, PPGN@DOX could be effectively internalized by 4T1 cells and could release DOX into the cell nuclei under hypoxic conditions. The in vitro anticancer results suggested that PPGN@DOX presented superior tumor cell-killing ability compared with free DOX in hypoxic environments. Furthermore, PPGN@DOX prolonged the blood circulation time and improved the biological distribution of DOX, resulting in increased antitumor outcomes and reduced side effects in vivo. Overall, the present work demonstrates that hypoxia-responsive nanoparticles have great application potential in the treatment of hypoxic tumors.

A PEGylated alternating copolymer with oxidation-sensitive phenylboronic ester pendants for anticancer drug delivery.

To target a response to a high oxidative stress environment of inflammatory or tumor sites, various reactive oxygen species (ROS) sensitive polymers have been developed as drug delivery systems. In this study, a novel oxidation sensitive copolymer, phenylboronic acid pinacol ester-functionalized methoxyl poly(ethylene glycol)-block-poly(phthalic anhydride-alter-glycidyl propargyl ether) (mPEG-b-P(PA-alt-GPBAe)), was designed and synthesized by ring-opening alternating copolymerization (ROAP) and click reaction. The copolymers could self-assemble into micelles in aqueous solution with an average size of 20.3 ± 9.3 nm, and are able to load hydrophobic anticancer drug (doxorubicin, DOX) with a high encapsulation efficiency of 75.2%. Interestingly, the encapsulated drug showed accelerated release in the trigger of H2O2, or at low pH values. The copolymers have low cytotoxicity indicated by the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay towards 4T1 cells, which showed cell viabilities of more than 80% with treatment of our copolymers at concentrations up to 0.5 mg mL-1. The effective uptake of the drug-loaded micelles by 4T1 cells was investigated by confocal laser scanning microscopy (CLSM) and flow cytometry (FCM) analysis. Finally, compared with free DOX, the DOX-loaded nanoparticles exhibited a better antitumor effect and had lower systemic toxicity in 4T1 tumor-bearing mice. Therefore, this new kind of copolymer acting as a stimuli-responsive nanocarrier should represent a promising therapeutic platform for cancer therapy.

Facile Synthesis of Resveratrol Nanogels with Enhanced Fluorescent Emission.

Herein, a kind of fluorescent resveratrol nanogels via one-pot thiol-ene Michael addition polymerization of resveratrol triacrylate, 1,6-hexanedithiol, and methoxyl poly(ethylene glycol) acrylate is prepared. The resultant nanogels can be well-dispersed in water with a hydrodynamic radius of around 68 nm, and the nanogels are stable in both water and organic solvents. Moreover, the resveratrol nanogels exhibit elevated fluorescence intensity compared to free resveratrol, and the quantum yield of resveratrol nanogels is estimated to be 5.8 times as that of free resveratrol dispersed in water. Fluorescence image results also demonstrate that the resveratrol nanogels can be used for cell imaging in MCF-7 human breast cancer cells. Therefore, the resveratrol nanogels are expected to be used as a trackable drug delivery system.

Au-aided reduced graphene oxide-based nanohybrids for photo-chemotherapy.

Graphene-based nanomaterials show great potential in photo-chemotherapy, but their photo-thermal effect is not very satisfactory. Herein, we presented a facile and low-cost strategy to grow Au clusters on the reduced graphene oxide (rGO) sheets aiming to improve photothermal effect. Au clusters with low-concentration was directly conjugated on the surface of rGO by electrostatic forces. To improve its biocompatibility, 3­(3­phenylureido) propanoic acid (PPA)-PEG (PPEG) had been introduced as biodegradable backbone to form rGO/Au/PPEG nanohybrids via π-π accumulation. The obtained rGO-based nanohybrids showed excellent biocompatibility, stability, low cytotoxicity, and enhanced photo-thermal conversion efficiency. To verify the synergistic photo-chemotherapy, doxorubicin (DOX) as a drug model had been loaded in rGO/Au/PPEG nanohybrids. The results indicated that rGO/Au/PPEG/DOX exhibited synergistic therapeutic efficacy compared with single chemotherapy or photothermal therapy, endowing this designed rGO-based nanohybrids with great potential for cancer treatments.

PEGylated Poly(α-lipoic acid) Loaded with Doxorubicin as a pH and Reduction Dual Responsive Nanomedicine for Breast Cancer Therapy.

Disulfide-containing nanoparticles are promising vehicles for anticancer drug delivery. However, the preparation of disulfide-containing nanoparticles usually relies on complex synthetic procedures. In the present work, a PEGylated poly(α-lipoic acid) (mPEG-PαLA) copolymer was facilely synthesized and used for pH and reduction dual responsive drug delivery. Poly(α-lipoic acid) was prepared by thermal polymerization of α-lipoic acid without any catalyst or solvent and then conjugated with methoxy poly(ethylene glycol) to form the mPEG-PαLA copolymer. The obtained mPEG-PαLA copolymer was amphiphilic, which could self-assemble into nanoparticles (NPs) in aqueous solution. More interestingly, the mPEG-PαLA NPs showed high drug loading efficiency (87.7%) for the cationic drug doxorubicin (DOX). The DOX-loaded NPs (NPs-DOX) exhibited pH and reduction dual responsive drug release behaviors. Moreover, the flow cytometry analysis and confocal laser scanning microscopy confirmed that the drug-loaded nanoparticles could be efficiently internalized and subsequently release DOX in 4T1 cancer cells. As a result, the NPs-DOX displayed favorable antiproliferation efficacy in 4T1 cancer cells (measured by MTT assays). Furthermore, the NPs-DOX showed enhanced antitumor efficacy in a 4T1 tumor-bearing mice model with reduced side toxicities toward normal organs due to the prolonged circulation time and improved biodistribution in vivo. In other words, this work demonstrates that the PEGylated poly(α-lipoic acid) copolymer can be used as a biocompatible and stimuli-responsive nanocarrier for anticancer drug delivery, which may have potential clinical utility.

Glutathione-triggered dual release of doxorubicin and camptothecin for highly efficient synergistic anticancer therapy.

An amphiphilic biodegradable prodrug (PLG-g-mPEG/CPT) was synthesized by conjugating disulfide-containing camptothecin (CPT) to poly(L-glutamic acid)-graft-methoxy poly(ethylene glycol) (PLG-g-mPEG) through esterification reaction. The amphiphilic prodrugs could self-assemble into micellar nanoparticles and encapsulate doxorubicin (DOX) in aqueous solution at pH 7.4. The treatment of the nanoparticles with reducing glutathione (GSH) at cytosolic concentration (10 mM) significantly promoted the in vitro dual release of DOX and CPT from the micelles. The results of flow cytometry (FCM) and confocal laser scanning microscopy (CLSM) manifested that the intracellular release of DOX and CPT from the micelles was enhanced by increasing the intracellular GSH level. Consistently, the MCF-7 cell killing mediated by the micelles was also intracellular GSH concentration-dependent. The low combination index (CI) value of < 0.3 demonstrated the high synergistic effect of DOX and CPT co-delivered by the nanoparticles in tumor cell killing. Therefore, this GSH-triggered dual release drug delivery system is a promising strategy for combination cancer therapy.

A glutathione-responsive sulfur dioxide polymer prodrug as a nanocarrier for combating drug-resistance in cancer chemotherapy.

Multidrug resistance (MDR) in cancer remains a significant challenge for curing cancer by chemotherapy. In this work, a kind of glutathione (GSH)-responsive polymer prodrug of SO2 was designed and synthesized, which presented synergistic effect with doxorubicin (DOX) for combating MCF-7 ADR human breast cancer cell. Firstly, a small molecular prodrug of SO2, N-(3-azidopropyl)-2,4-dinitrobenzenesulfonamide (AP-DNs), was chemically conjugated onto the side chain of methoxy poly (ethylene glycol)-block-poly (γ-propargyl-l-glutamate) (mPEG-PPLG) block copolymer to generate an amphiphilic polymer prodrug of SO2, mPEG-PLG (DNs). The obtained mPEG-PLG (DNs) prodrug could self-assemble into micelles in aqueous media and release SO2 rapidly in response to thiol compounds. Then, DOX was loaded into mPEG-PLG (DNs) nanoparticles with ultrahigh drug-loading efficiency (97.3%). In vitro drug release tests indicated that the DOX-loaded nanoparticles could simultaneously release SO2 and DOX by GSH triggering. Moreover, the effective cellular uptake of the DOX-loaded nanoparticles and subsequent intracellular release of SO2 and DOX were verified by confocal laser scanning microscopy (CLSM) and flow cytometry (FCM) analyses. The released SO2 could promote the reactive oxygen species (ROS) level in tumor cells, which thereby resulted in oxidative damages of cancer cells, together with restoration of MCF-7 ADR cells sensitivity to DOX. As a result, the released DOX and SO2 showed synergistic therapeutic effect against MCF-7 ADR cells. In vivo antitumor evaluation further indicated that, compared with free DOX, the DOX-loaded nanoparticles exhibited better antitumor effect in a MCF-7 ADR-xenografted nude mice model while had lower system toxicity. Overall, we demonstrated, for the first time, that a SO2 polymer prodrug, acting as a stimuli-responsive nanocarrier to codeliver DOX, can efficiently inhibit the proliferation of MDR tumor cells, which may offer a new weapon for combating MDR in cancer therapy.

Rapid fluorescence imaging of spinal cord following epidural administration of a nerve-highlighting fluorophore.

Iatrogenic spinal cord injury (SCI) is the most devastating complication of spine surgery, which usually results in permanent and serious disabilities of patients. Improvement of the visualization and discrimination of the spinal cord is critical for accuracy and safety during surgery; however, to date, there is no suitable technology to fulfill this clinical need. Here, we first show an efficient and rapid fluorescence imaging of the spinal cord in rabbit by epidural administration of a nerve-highlighting fluorophore, i.e. (E, E)-1,4-bis(p-aminostryl)-2-methoxy benzene (BMB). The BMB is firstly encapsulated into polymeric micelles to form a BMB-micelle (BMB-m) formulation with well-dispersion in normal saline solution. After epidural administration of BMB-m, BMB is transported by the flow of cerebrospinal fluid (CSF) and binds to the peripheral region of the white matter thus facilitating rapid staining of the spinal cord. Furthermore, this BMB imaging technology also holds great potential for visually monitoring the integrity of the spinal cord in real time and promptly identifying acute SCI during spine surgery.