Cancer Cell-targeted Drug Delivery Research Articles

Antibody-mimetic drug conjugate with efficient internalization activity using anti-HER2 VHH and duocarmycin

Antibody-mimetic drug conjugate (AMDC) is a cancer cell-targeted drug delivery system based on the non-covalent binding of mutated streptavidin and modified biotin, namely Cupid and Psyche. However, the development of AMDCs is hampered by difficulties in post-translational modification or poor internalization activity. Here, we report an expression, refolding, and purification method for AMDC using a variable heavy chain of heavy chain-only antibodies (VHHs). Monomeric anti-HER2 VHH fused to Cupid was expressed in Escherichia coli inclusion bodies. Solubilization and refolding at optimized reducing conditions and pH levels were selected to form a functional, tetrameric protein (anti-HER2 VHH-Cupid) that can be easily purified based on molecular weight. Anti-HER2 VHH-Cupid non-covalently creates a tight complex with Psyche linked to a potent DNA-alkylating agent, duocarmycin. This complex can be absorbed by the HER2-expressing human breast cancer cell line, KPL-4, and kills KPL-4 cells in vitro and in vivo. The production of a targeting protein with internalizing activity, combined with the non-covalent conjugation of a highly potent payload, renders AMDC a promising platform for developing cancer-targeted therapy.

Recent advances in the development of biocompatible nanocarriers and their cancer cell targeting efficiency in photodynamic therapy.

In recent years, the role of biocompatible nanocarriers (BNs) and their cancer cell targeting efficiency in photodynamic therapy (PDT) holds potential benefits for cancer treatment. Biocompatible and biodegradable nanoparticles are successfully used as carrier molecules to deliver cancer drugs and photosensitizers due to their material safety in the drug delivery system. Biocompatible nanocarriers are non-toxic and ensure high-level biocompatibility with blood, cells, and physiological conditions. The physicochemical properties of BNs often enable them to modify their surface chemistry, which makes conjugating specific ligands or antibodies to achieve cancer cell targeting drug delivery in PDT. This review article focuses on the various types of BNs used in targeted drug delivery, physicochemical properties, and surface chemistry of BNs in targeted drug delivery, advantages of BNs in drug delivery systems, and the targeting efficiency of BNs on some specific targeting receptors for cancer therapy. Furthermore, the review briefly recaps the nanocarrier-based targeted approaches in cancer PDT.

Insights into Tumor Microenvironment (TME) and the Nano Approaches to Suppress Tumor Growth

Cancer, the second major cause of death after cardiovascular disease has been reported to have resulted in over 10 million deaths worldwide per year, on average. Resistance to therapeutic interventions is a definitive characteristic of cancer, where acquired resistance has become a challenge that has significantly reduced the anti-tumor efficiency of cancer therapy. The tumor microenvironment (TME) is heavily responsible for these cancer cell-targeted drug delivery system challenges due to its abundant physiological and pathological barriers that impair the tumoral penetration and distribution of nanotherapeutics. However, these unique TME characteristics also make the TME an attractive therapeutic target and a focus of intense research in the development of antitumor nanomedicine strategies that can directly target or modulate it. In this review, we summarize the strategies developed to target the TME to overcome the obstacles against nano-drug delivery and consequently improve anti-tumor efficiency.

Chimeric virus-like particles (VLPs) designed from shrimp nodavirus (MrNV) capsid protein specifically target EGFR-positive human colorectal cancer cells

Recombinant MrNV capsid protein has been shown to effectively deliver plasmid DNA and dsRNA into Sf9 insect cells and shrimp tissues. To extend its application to cancer cell-targeting drug delivery, we created three different types of chimeric MrNV virus-like particles (VLPs) (R-MrNV, I-MrNV, and E-MrNV) that have specificity toward the epidermal growth factor receptor (EGFR), a cancer cell biomarker, by incorporating the EGFR-specific GE11 peptide at 3 different locations within the host cell recognition site of the capsid. All three chimeric MrNV-VLPs preserved the ability to form a mulberry-like VLP structure and to encapsulate EGFP DNA plasmid with an efficiency comparable to that previously reported for normal MrNV (N-MrNV). Compared to N-MrNV, the chimeric R-MrNV and E-MrNV carrying the exposed GE-11 peptide showed a significantly enhanced binding and internalization abilities that were specific towards EGFR expression in colorectal cancer cells (SW480). Specific targeting of chimeric MrNV to EGFR was proven by both EGFR silencing with siRNA vector and a competition with excess GE-11 peptide as well as the use of EGFR-negative colorectal cells (SW620) and breast cancer cells (MCF7). We demonstrated here that both chimeric R-MrNV and E-MrNV could be used to encapsulate cargo such as exogenous DNA and deliver it specifically to EGFR-positive cells. Our study presents the potential use of surface-modified VLPs of shrimp virus origin as nanocontainers for targeted cancer drug delivery.

Cancer Cell Preferential Penetration and pH-Responsive Drug Delivery of Oligorutin.

We report herein a novel delivery system, derived from the facile enzymatic synthesis of oligorutin (OR), for cancer cell targeting and pH-responsive drug delivery. In this study, we demonstrate that OR could preferentially penetrate cancer cells via the lipid raft-mediated endocytosis pathway, and cell membrane cholesterol was critical to the internalization of OR. The accumulation of OR in the tumor region was further confirmed by an in vivo biodistribution study. Considering the tumor-targeting property of OR, a pH-responsive drug delivery system (OR-BTZ) was developed by covalent conjugation of the catechol groups on OR with antitumor drug bortezomib (BTZ) through a pH-sensitive borate ester bond. OR-BTZ exerted cytotoxicity as well as inhibition of the migration and invasion to hepatoma carcinoma cells and showed no apparent cytotoxicity with liver normal cells. The OR-BTZs also presented significant therapeutic efficacy and low systematic toxicity in the murine hepatocellular carcinoma model. To our knowledge, this study presents the first attempt to exploit the potential of oligoflavonoids for cancer cell-targeted drug delivery and will motivate the development of flavonoids and their derivatives as a new type of biomaterials for tumor-targeted therapy.

Magnetic particle anchored reduction and pH responsive nanogel for enhanced intracellular drug delivery

The controlled delivery of drugs towards the cancerous cell is still an unresolved issue in the area of cancer cell targeted drug delivery. To resolve the issue, in this work, a triple-responsive (pH, magnetic field and redox) nanogel has been fabricated to potentially deliver the anticancer drug (DOX) to the cancerous cell. The nanogel has been fabricated by esterification reaction between the ‘-COOH’ group of the pre-synthesized amphiphilic tetra-armed block copolymer, pentaerythritol-poly(ε-caprolactone)-b-polyacrylic (Pe-PCL-b-PAA) and ‘-OH’ group of the redox-responsive crosslinker, 2-hydroxyethyl disulfide. The nanogels are further ornamented with aminosilane coated magnetic nanoparticles to impart the magnetic field responsive characteristic. The spherically shaped nanogel (size 120 nm) ensures the long circulation half-life in the bloodstream. The drug release profile under different stimuli such as pH, magnetic field and redox environment explored a new way of controlled drug release in a more promising manner under the synergistic participation of all three stimuli. It has been observed that the multi-stimuli responsive nanogel not only boost up the cell uptake process but also dramatically enhance the drug release rate up to 65% within 1 h under the environment of multiple stimuli (acidic pH 5, redox (10 mM GSH) and in the presence of dynamic magnetic field. Precisely, the acidic pH and reducing micro-environment of cancer affected cells are expected to act as an internal stimulus, whereas magnetic force can be maneuverer as an external stimulus in the proposed sub-structure of the nanogels. In a nut-shell, this kind of multi-responsive nanogel can pave a new direction in the area of controlled and targeted delivery of anticancer drugs or other biomolecules.

Galactosylated chitosan-functionalized mesoporous silica nanoparticles for efficient colon cancer cell-targeted drug delivery.

Targeted drug delivery to colon cancer cells can significantly enhance the therapeutic efficiency. Herein, we developed 5-fluorouracil (5-FU)-loaded amino-functionalized mesoporous silica nanoparticle (MSN-NH2)-based galactosylated chitosans (GCs), which are galactose receptor-mediated materials for colon-specific drug delivery systems. Both unmodified and functionalized nanoparticles were characterized by scanning electron microscopy, transmission electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, nitrogen sorption and dynamic light scattering. Drug loading capacity and drug release properties were determined by ultraviolet spectrophotometry. 5-FU@MSN-NH2/GC showed high loading capacity and possessed much higher cytotoxicity on human colon cancer cells (SW620 cells) than 5-FU@MSN-NH2 and free 5-FU. But, MSN-NH2/GC did not show significant cytotoxicity. Subsequently, 5-FU@MSN-NH2/GC anti-cancer activity on SW620 cells in vitro was confirmed by cell apoptosis. These results are consistent with the cellular uptake test in which MSN-NH2/GC could specifically recognize and bind to cancer cells by the galectin-receptor recognition. But, it is found that pre-addition of galactose in the medium, leading to competitive binding to the galectin receptor of SW620 cells, resulted in a decrease in the binding of MSN-NH2/GC to the galectin receptor. The results demonstrated the inorganic–organic nanocomposite could be used as a promising drug delivery carrier for the targeted delivery of drug into galectin-positive colon cancer cells to improve therapeutic index while reducing side effects.

Galactosylated Chitosan-Functionalized Mesoporous Silica Nanoparticle Loading by Calcium Leucovorin for Colon Cancer Cell-Targeted Drug Delivery.

Targeted drug delivery to colon cancer cells can significantly improve the efficiency of treatment. We firstly synthesized carboxyl-modified mesoporous silica nanoparticles (MSN–COOH) via two-step synthesis, and then developed calcium leucovorin (LV)-loaded carboxyl-modified mesoporous silica nanoparticles based on galactosylated chitosan (GC), which are galectin receptor-mediated materials for colon-specific drug delivery systems. Both unmodified and functionalized nanoparticles were characterized by scanning electron microscopy (SEM), transmission electron microscope (TEM), X-ray diffraction (XRD), Fourier transform infrared (FT-IR), nitrogen sorption, and dynamic light scattering (DLS). Drug release properties and drug loading capacity were determined by ultraviolet spectrophotometry (UV). LV@MSN–COOH/GC had a high LV loading and a drug loading of 18.07%. In vitro, its release, mainly by diffusion, was sustained release. Cell experiments showed that in SW620 cells with the galectin receptor, the LV@MSN–COOH/GC metabolized into methyl tetrahydrofolic acid (MTHF) and 5-fluorouracil (5-FU)@MSN–NH2/GC metabolized into FdUMP in vivo. MTHF and 5-fluoro-2′-deoxyuridine 5′-monophosphate (FdUMP) had combined inhibition and significantly downregulated the expression of thymidylate synthase (TS). Fluorescence microscopy and flow cytometry experiments show that MSN–COOH/GC has tumor cell targeting, which specifically recognizes and binds to the galectin receptor in tumor cells. The results show that the nano-dosing system based on GC can increase the concentrations of LV and 5-FU tumor cells and enhance their combined effect against colon cancer.

Mesoporous Carbon Nanospheres with ZnO Nanolids for Multimodal Therapy of Lung Cancer.

To improve therapeutic efficacy, we designed novel poly(ethylene glycol) (PEG)-modified oxidized mesoporous carbon nanospheres (OMCNPs) that were loaded with the therapeutic drug doxorubicin (DOX) and targeted with ligand hyaluronic acid (HA) to kill lung cancer cells. ZnO quantum dots (ZnO QDs) were introduced not only to cap OMCNP-based carriers but also to chelate with DOX, which exhibited a high drug-loading capacity. Upon cellular uptake, the pH-sensitive ZnO lids dissolved to Zn2+ in tumor cells, resulting in dissociation of the Zn2+-DOX complex and controlled release of DOX. Moreover, the novel OMCNP-based system could generate hyperthermia and facilitate the release of loaded DOX with near-infrared (NIR) irradiation. The combined chemo-/photothermal-targeted therapy exhibited significant advantages superior to single chemotherapy or photothermal therapy. This technology provided an excellent lung cancer cell-targeted drug delivery system, integrating chemotherapy with photothermal therapy. This chemo-/photothermal therapy effectively improved the therapeutic efficacy with lower adverse effects to normal tissues.

Functionalized chitosan with super paramagnetic hybrid nanocarrier for targeted drug delivery of curcumin

Recently, hydrophobically functionalized polymers have been deployed as carriers to improve the encapsulation of hydrophobic drugs. The metal nanocomposites are extensively used to improve the biocompatibility of the formulation and target the drug to the specialized site. In our current study, naphthalene acetate (NAA) was incorporated into the amine group of chitosan to form a hydrophobically functionalized chitosan–NAA drug delivery carrier. The calcium ferrite nanoparticles (CFNP) were embedded in the chitosan–NAA matrix to form a super paramagnetic hybrid nanocarrier for controlled curcumin drug delivery. Various analytical techniques were performed to ensure the functional group modifications, thermal stability, surface nature and morphological behavior of synthesized hybrid carriers. The maximum encapsulation efficiency of 93.6% was obtained under the optimized conditions of drug to chitosan–NAA at 0.1, CFNP to chitosan–NAA at 0.75 and TPP to chitosan–NAA at 1.0 (w/w) ratios, respectively, by adapting Taguchi method. Drug release studies were conducted to determine the effect of pH, drug loading concentrations and magnetic field responses. The drug release data were fitted to various kinetic release models to understand the drug release mechanism. The biocompatibility of the hybrid material was tested using L929 mouse fibroblast cells. The cytotoxicity test against breast cancer cells (MCF-7) was also performed to study the anticancer property of the hybrid paramagnetic material. The prepared curcumin-loaded chitosan–NAA/CFNP was very active against cancer cells in comparison to the normal cells. The results confirmed the applicability of the hybrid nanocarriers in cancer cell-targeted drug delivery.

PH and near-infrared active; chitosan-coated halloysite nanotubes loaded with curcumin-Au hybrid nanoparticles for cancer drug delivery

In this work, we propose biofriendly in-situ preparation method of Au NPs (hexagonal and rod-shape structures) in the lumen as well as the surface cage of biocompatible halloysite nanotubes (HNTs) using curcumin (CUR) as anticancer drug and subsequently coating with bio-adhesive chitosan (CS) as a polysaccharide. The formation of Au NPs and their interactions with CUR and CS exist in the HNTs has been characterized by FTIR, XRD, XPS, STEM techniques. Interestingly, Au NPs showed longitudinal plasmon resonance bands at 760 and 980 nm that indicate the near-infrared (NIR) responsive property of hybrid nanoparticles. Rod shape and hexagonal structures of Au NPs were produced as confirmed by TEM images. The loading efficiency of CUR was found as much as 12%. Importantly, more CUR release was achieved under acidic conditions (pH 5.5) than basic conditions (pH 7.4). The anticancer potential of HNT hybrid nanoparticles on MCF-7 cancer cells was studied and showed efficient anticancer activity under intracellular tumor cell environment (pH 5.5) than extracellular conditions (pH 7.4). Moreover, the developed HNT hybrid nanoparticles consisting of Au NPs (NIR responsive property) and pH-responsive CUR release could make it suitable for cancer cell-targeted drug delivery platform with NIR-imaging.

Engineering a customized nanodrug delivery system at the cellular level for targeted cancer therapy.

Drug administration customized to individual cells could intrinsically address cancer heterogeneity and provide a safe and effective method for delivering personalized treatment. To accomplish this, we developed a smart nanodrug delivery system characterized by cancer cell-targeted drug delivery and intracellular biomarker-responsive drug activation. This system was composed of a long-nicked DNA duplex formed by tandem hybridization of two extended antisense oligonucleotides whose ends were separately blocked with a cancer cell-specific aptamer, AS1411, and a replaceable anti-biomarker probe (ABP). We demonstrated that this DNA nanodrug was directed to cancer cells with the guidance power of AS1411 and then activated by the presence of a given intracellular biomarker. By using such a belt-and-braces strategy, this DNA nanodrug system could safely and efficiently accelerate apoptosis of target cancer cells. Moreover, since the expression level of biomarkers tends to indicate the specific physiological state of individual cells, biomarker-responsive activation of the nanodrug is expected to enable customized drug administration at the cellular level.

Circulating GRP78 antibodies from ovarian cancer patients: a promising tool for cancer cell targeting drug delivery system?

Glucose-regulated protein 78 (GRP78) is a chaperone protein that has a high frequency in tumor cells. Normally it is found in the endoplasmic reticulum to assist in protein folding, but under cellular stress, GRP78 influences proliferative signaling pathways at the cell surface. The increased expression elicits autoantibody production, providing a biomarker of ovarian cancer, as well as other types of cancer. This study aims to determine the epitope recognition of GRP78 autoantibodies isolated from serum of ovarian cancer patients and use the identified antibodies to design new drug delivery systems to specifically target cancer cells. We first confirmed that the membrane GRP78 levels are increased in ovarian cancer cells and positively correlate with proliferation. However, the level of circulating GRP78 autoantibodies did not correlate with membrane GRP78 expression in ovarian cancer cells and was lower, although not significantly, compared to control patients. We then determined the epitope recognition of GRP78 autoantibodies and showed that treatment with paclitaxel-loaded nanoparticles coated with anti-GRP78 antibodies significantly decreased tumor development in chick embryo culture of ovarian cancer cell tumors compared to paclitaxel treatment alone. This evidence suggests that nanoparticle drug delivery systems coupled with antibodies against GRP78 has potential as a powerful therapy against ovarian cancer.

Graphene oxide and adenosine triphosphate as a source for functionalized carbon dots with applications in pH-triggered drug delivery and cell imaging

A folate-functionalized carbon dot-based nanocarrier system has been successfully synthesized for cancer cell targeted drug delivery.

Remotely controlled red blood cell carriers for cancer targeting and near-infrared light-triggered drug release in combined photothermal chemotherapy

Red blood cells (RBCs), the “innate carriers” in blood vessels, are gifted with many unique advantages in drug transportation over synthetic drug delivery systems (DDSs). Herein, a tumor angiogenesis targeting, light stimulus-responsive, RBC-based DDS is developed by incorporating various functional components within the RBC platform. An albumin bound near-infrared (NIR) dye, together with a chemotherapy drug doxorubicin, is encapsulated inside RBCs, the surfaces of which are modified with a targeting peptide to allow cancer targeting. Under stimulation by an external NIR laser, the membrane of the RBCs would be destroyed by the light-induced photothermal heating, resulting in effective drug release. As a proof of principle, RBC-based cancer cell targeted drug delivery and light-controlled drug release is demonstrated in vitro, achieving a marked synergistic therapeutic effect through the combined photothermal–chemotherapy. This work presents a novel design of smart RBC carriers, which are inherently biocompatible, promising for targeted combination therapy of cancer.

EpCAM aptamer-functionalized mesoporous silica nanoparticles for efficient colon cancer cell-targeted drug delivery

Targeted delivery of anticancer agents by functional nanoparticles is an attractive strategy to increase their therapeutic efficacy while reducing toxicity. In this work, doxorubicin (DOX)-loaded mesoporous silica nanoparticles (MSNs) were modified with aptamer (Ap) against the epithelial cell adhesion molecule (EpCAM) for targeted delivery of DOX to colon cancer cells. These nanoparticles (Ap-MSN-DOX) were characterized by particle size, zeta potential, aptamer conjugation efficiency, drug encapsulation efficiency, and drug release properties. The in vitro cell recognition, cellular uptake, EpCAM protein inhibition efficiency, and cytotoxicity of Ap-MSN-DOX were also studied. Results demonstrated that EpCAM conjugation increased binding of Ap-MSN-DOX to EpCAM over-expressing SW620 colon cancer cells but not EpCAM-negative Ramos cells, resulting in enhanced cellular uptake and increased cytotoxicity of the DOX in SW620 cells when compared to non-Ap-modified nanoparticles (MSN-DOX). Additionally, Ap-MSN-DOX exhibited significant inhibition effects on the expression of EpCAM on SW620 cells. These results suggested that Ap-MSN-DOX has the potential for the targeted delivery of therapeutic agents into EpCAM positive colon cancer cells to improve therapeutic index while reducing side effects.

Radiosensitivity enhancement of radioresistant glioblastoma by epidermal growth factor receptor antibody-conjugated iron-oxide nanoparticles.

The epidermal growth factor receptor deletion variant EGFRvIII is known to be expressed in a subset of patients with glioblastoma (GBM) tumors that enhances tumorigenicity and also accounts for radiation and chemotherapy resistance. Targeting the EGFRvIII deletion mutant may lead to improved GBM therapy and better patient prognosis. Multifunctional magnetic nanoparticles serve as a potential clinical tool that can provide cancer cell targeted drug delivery, imaging, and therapy. Our previous studies have shown that an EGFRvIII-specific antibody and cetuximab (an EGFR- and EGFRvIII-specific antibody), when bioconjugated to IONPs (EGFRvIII-IONPs or cetuximab-IONPs respectively), can simultaneously provide sensitive cancer cell detection by magnetic resonance imaging (MRI) and targeted therapy of experimental GBM. In this study, we investigated whether cetuximab-IONPs can additionally allow for the radiosensitivity enhancement of GBM. Cetuximab-IONPs were used in combination with single (10Gy×1) or multiple fractions (10Gy×2) of ionizing radiation (IR) for radiosensitization of EGFRvIII-overexpressing human GBM cells in vitro and in vivo after convection-enhanced delivery (CED). A significant GBM antitumor effect was observed in vitro after treatment with cetuximab-IONPs and subsequent single or fractionated IR. A significant increase in overall survival of nude mice implanted with human GBM xenografts was found after treatment by cetuximab-IONP CED and subsequent fractionated IR. Increased DNA double strands breaks (DSBs), as well as increased reactive oxygen species (ROS) formation, were felt to represent the mediators of the observed radiosensitization effect with the combination therapy of IR and cetuximab-IONPs treatment.

Remotely Controlled Red Blood Cell Carriers for Cancer Targeting and Near‐Infrared Light‐Triggered Drug Release in Combined Photothermal–Chemotherapy

Red blood cells (RBCs), the “innate carriers” in blood vessels, are gifted with many unique advantages in drug transportation over synthetic drug delivery systems (DDSs). Herein, a tumor angiogenesis targeting, light stimulus‐responsive, RBC‐based DDS is developed by incorporating various functional components within the RBC platform. An albumin bound near‐infrared (NIR) dye, together with a chemotherapy drug doxorubicin, is encapsulated inside RBCs, the surfaces of which are modified with a targeting peptide to allow cancer targeting. Under stimulation by an external NIR laser, the membrane of the RBCs would be destroyed by the light‐induced photothermal heating, resulting in effective drug release. As a proof of principle, RBC‐based cancer cell targeted drug delivery and light‐controlled drug release is demonstrated in vitro, achieving a marked synergistic therapeutic effect through the combined photothermal–chemotherapy. This work presents a novel design of smart RBC carriers, which are inherently biocompatible, promising for targeted combination therapy of cancer.

Multifunctional pH-Disintegrable micellar nanoparticles of asymmetrically functionalized β-cyclodextrin-Based star copolymer covalently conjugated with doxorubicin and DOTA-Gd moieties

We report on a novel type of multifunctional pH-disintegrable micellar nanoparticles fabricated from asymmetrically functionalized β-cyclodextrin ( β-CD) based star copolymers covalently conjugated with doxorubicin (DOX), folic acid (FA), and DOTA- Gd moieties for integrated cancer cell-targeted drug delivery and magnetic resonance (MR) imaging contrast enhancement. Asymmetrically functionalized β-CD, ( N 3) 7- CD-( Br) 14, which possesses 7 azide functionalities and 14 α-bromopropionate moieties in the upper and lower rim of rigid toroidal β-CD core, respectively, was synthesized at first. The subsequent atom transfer radical polymerization (ATRP) of N-(2-hydroxypropyl) methacrylamide (HPMA), conjugation with DOX and FA, and click reaction with alkynyl-(DOTA- Gd) complex afforded (DOTA- Gd) 7- CD-(PHPMA- FA- DOX) 14 star copolymer comprising of 7 DOTA- Gd complex moieties and 14 PHPMA arms covalently anchored with DOX and FA via acid-labile carbamate linkages and ester bonds, respectively. The covalent conjugation with ∼13 DOX molecules onto PHPMA arms per star copolymer (∼14 wt% loading content) endows the initially hydrophilic one with amphiphilicity, leading to the self-assembly into micellar nanoparticles of several tens of nanometers in aqueous solution at pH 7.4. In vitro DOX release profile from micellar nanoparticles is highly pH-dependent, and over a time period of 42 h, cumulative releases of ∼10%, 53%, and 89% conjugated DOX at pH 7.4, 5.0, and 4.0, respectively, were observed. Most importantly, the pH-modulated release of conjugated DOX from micellar nanoparticles is accompanied with the micelle disintegration due to the loss of amphiphilicity of the star copolymer scaffold. In vitro cell viability assays revealed that (DOTA -Gd) 7- CD-(PHPMA 15) 14 star copolymer is almost non-cytotoxic up to a concentration of 0.5 g/L, whereas DOX-conjugated micellar nanoparticles of (DOTA- Gd) 7- CD-(PHPMA- FA- DOX) 14 can effectively enter and kill HeLa cells at a concentration higher than ∼80 mg/L. In vitro MR imaging experiments indicated a considerably enhanced T 1 relaxivity ( r 1 = 11.4 s −1 mM −1) for micellar nanoparticles compared to that for the small molecule counterpart, alkynyl-DOTA-Gd ( r 1 = 3.1 s −1 mM −1). In vivo MR imaging assay in rats revealed considerable accumulation of micellar nanoparticles within rat liver and kidney and prominent positive contrast enhancement. The integrated design of diagnostic and therapeutic functions of multifunctional pH-disintegrable micellar nanoparticles augurs well for their practical applications in the field of image-guided cancer chemotherapy.

Folate-Functionalized Unimolecular Micelles Based on a Degradable Amphiphilic Dendrimer-Like Star Polymer for Cancer Cell-Targeted Drug Delivery

A folate-functionalized degradable amphiphilic dendrimer-like star polymer (FA-DLSP) with a well-defined poly(L-lactide) (PLLA) star polymer core and six hydrophilic polyester dendrons based on 2,2-bis(hydroxymethyl) propionic acid was successfully synthesized to be used as a nanoscale carrier for cancer cell-targeted drug delivery. This FA-DLSP hybrid formed unimolecular micelles in the aqueous solution with a mean particle size of ca. 15 nm as determined by dynamic light scattering and transmission electron microscopy. To study the feasibility of FA-DLSP micelles as a potential nanocarrier for targeted drug delivery, we encapsulated a hydrophobic anticancer drug, doxorubicin (DOX), in the hydrophobic core, and the loading content was determined by UV-vis analysis to be 4 wt %. The DOX-loaded FA-DLSP micelles demonstrated a sustained release of DOX due to the hydrophobic interaction between the polymer core and the drug molecules. The hydrolytic degradation in vitro was monitored by weight loss and proton nuclear magnetic resonance spectroscopy to gain insight into the degradation mechanism of the FA-DLSP micelles. It was found that the degradation was pH-dependent and started from the hydrophilic shell gradually to the hydrophobic core. Flow cytometry and confocal microscope studies revealed that the cellular binding of the FA-DLSP hybrid against human KB cells with overexpressed folate-receptors was about twice that of the neat DLSP (without FA). The in vitro cellular cytotoxicity indicated that the FA-DLSP micelles (without DOX) had good biocompatibility with KB cells, whereas DOX-loaded micelles exhibited a similar degree of cytotoxicity against KB cells as that of free DOX. These results clearly showed that the FA-DLSP unimolecular micelles could be a promising nanosize anticancer drug carrier with excellent targeting property.