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A Dual-Responsive Bola-Type Supra-Amphiphile Constructed from Water-Soluble Pillar[5]arene and NaphthalimideContaining Amphiphile for Intracellular Drug Delivery Xin Liu, Keke Jia, Yichen Wang, Wei Shao, Chenhao Yao, Luming Peng, Dongmei Zhang, Xiao-Yu Hu, and Leyong Wang ACS Appl. Mater. Interfaces, Just Accepted Manuscript • DOI: 10.1021/acsami.7b00643 • Publication Date (Web): 18 Jan 2017 Downloaded from http://pubs.acs.org on January 21, 2017

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ACS Applied Materials & Interfaces

A Dual-Responsive Bola-Type Supra-Amphiphile Constructed from Water-Soluble Pillar[5]arene and Naphthalimide-Containing Amphiphile for Intracellular Drug Delivery Xin Liu,†∥ Keke Jia,‡∥ Yichen Wang,† Wei Shao,† Chenhao Yao,† Luming Peng,† Dongmei Zhang,*‡ Xiao-Yu Hu,*† and Leyong Wang†§ †

Key Laboratory of Mesoscopic Chemistry of MOE and Collaborative Innovation Center of

Chemistry for Life Sciences, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, China. ‡

State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing

University, Nanjing, 210023, China §

Jiangsu Key Laboratory of Advanced Catalytic Materials & Technology, School of

Petrochemical Engineering, Changzhou University, Changzhou, 213164, China

KEYWORDS. Dual-responsiveness, Supra-amphiphile, Pillar[5]arene, Self-assembly, Drug Delivery

ACS Paragon Plus Environment

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ABSTRACT. Supramolecular construction of multi-stimuli platform for drug delivery is a challenging task. In this work, a pH and GSH (glutathione) dual-responsive bola-type supramolecular amphiphile was successfully fabricated by the complexation between a watersoluble pillar[5]arene (WP5) and a bolaform naphthalimide guest (G) in water. The resulting bola-type amphiphile further self-assembled into supramolecular binary vesicles, which could be disassembled by low pH and/or high GSH concentration environment. Furthermore, the results of drug loading and releasing tests showed that doxorubicin (DOX), the hydrophobic anticancer drug, could be successfully encapsulated into the Stern region of the obtained supramolecular vesicles and generated the DOX-loaded vesicles with good drug-loading efficiency. Moreover, the obtained DOX-loaded vesicles displayed efficient and rapid DOX release at a simulated tumor microenvironment with low-pH and/or excess GSH conditions. Significantly, cytotoxicity experiments revealed that the DOX-loaded supramolecular vesicles could obviously improve the anticancer efficiency of free DOX for tumor cells, while remarkably reduce its side effects for normal cells. In vitro cellular uptaking and subcellular localization assays further proved that these smart drug nanovehicles, entering cancer cells mainly via endocytosis, could cause excellent drug accumulation in cancer cells. The present study provides a successfully example to rational design an effective bola-type stimuli-responsive supramolecular nanocarrier, which might have wide potential applications in constructing various controlled drug delivery systems.


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Introduction Although considerable efforts have been made in cancer therapy, traditional chemotherapy is still the most effective treatment for malignant tumor.1 However, it also kills healthy cells and causes severe side effects to patients, mainly due to the nonspecific tissue biodistribution of chemotherapeutic drugs.2 Moreover, only a small portion of drugs can reach the tumor tissues because of the systemic distribution, which leads to the relatively low drug efficacy.3 To solve these problems, fully using nanotechnology for drug delivery should be taken into consideration, which will bring new attractions for cancer therapy.4-9 Nanocarrier-based drug delivery systems have unique properties that allow for both passive and active targeting of malignant tumors through the enhanced permeation and retention (EPR) effect, which can facilitate the efficient accumulation of drug nanocarriers especially in tumor sites.10-13 In addition, a nanocarrier system incorporated with stimuli-responsive properties (such as a higher GSH concentration, a lowered pH microenvironment, or an increased level of certain enzymes in cancer cells), could be liable to overcome some of the systemic or intracellular delivery barriers and released the therapeutic agents at pathological sites.14-18 Supramolecular amphiphiles containing both hydrophilic and hydrophobic portions held together via non-covalent interactions, offer new opportunities for the fabrication of smart drug delivery systems (DDSs).19-24 As compared to traditional amphiphiles, supramolecular amphiphiles provide easy and facile approaches for building well-defined advanced structures, which could effectively avoid multiple synthesis steps and complicated purification processes during the fabrications.25-28 Furthermore, due to the non-covalent linkage by relatively weak and dynamic interactions, supramolecular amphiphiles have the special ability to undergo reversible switching of structures, morphologies, and functions in response to certain external stimuli, such


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as pH,29-30 temperature,31, 32 light,33, 34 enzyme,35, 36 and redox agents,37, 38 which make them with great potential applications in drug delivery. Therefore, it provides a robust and flexible platform for constructing functional and intelligent supramolecular nanovehicles, which are urgently needed for modern nanomedicine. Pillar[n]arenes, an emerging type of macrocyclic hosts, have received great attentions due to their symmetrical pillar and unique rigid architectures, easy functionalization,39-42 and excellent abilities to selectively bind with various kinds of guest molecules.43-50 Based on the great efforts made by chemists and materials scientists, numerous pillar[n]arene-based dynamic host-guest recognition motifs have been built and further applied in the fabrication of various functional materials, including mechanically interlocked molecules,51,52 chemosensors,53,54 artificial transmembrane channels,55,56 and especially DDSs.57-62 As macrocyclic host molecules, watersoluble pillar[5,6]arenes (WP5, WP6) were usually used to construct supramolecular nanovehicles because of their excellent pH-responsive capabilities and solubility in water.63,64 Additionally, it is well known that the sensitivity of disulfide bond toward GSH is very important for constructing stimuli-responsive62 anticancer drug delivery systems due to the particular low pH condition and much higher GSH concentration in the tumor microenvironment.65 Furthermore, naphthalene imides (NDI) are very attractive building blocks since they possess extended π systems for effectively improving the stability of assembly and they also belong to an important class of excellent DNA-binding agents.66 Therefore, we want to design a disulfide bond containing bolaform naphthalimide guest molecule with two quaternary ammonium groups in both terminals, which could form a bola-type and dual-responsive supramolecular amphiphile with pH-responsive water-soluble pillar[5]arenes (WP5), and it could be used to advanced drug delivery systems.


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Herein, we demonstrated a bola-type pillar[5]arene-based pH and GSH dual-responsive supramolecular drug delivery system, which was constructed by a water-soluble pillar[5]arene (WP5) and a bolaform naphthalimide derivative (G) bearing disulfide bonds on both sides as depicted in Scheme 1. The bola-type supra-amphiphile WP5⊃ ⊃G could further form hollow supramolecular vesicles, which could effectively encapsulate doxorubicin (DOX), hydrophobic anticancer drug, in the Stern region. Moreover, the loaded drug could be released effectively in a simulated tumor microenvironment with acidic and high glutathione concentration. Notably, cytotoxicity experiments indicated that DOX-loaded vesicles could not only significantly enhance the anticancer efficiency of DOX, but also effectively reduce the side effects to normal tissue and cells compared with free DOX. Cellular uptaking and subcellular localization measurements further revealed that this kind of supramolecular drug nanovehicle, entering cancer cells mainly through endocytosis, which could result in a remarkable drug accumulation especially in tumor cells, indicating its potential applications for cancer treatment.


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Scheme 1. Schematic illustration of constructing bola-type supramolecular vesicles and their properties in efficient drug delivery.

Results and Discussion Investigation of the Host-Guest Complexation. WP5 was synthesized in accordance with the reported procedure,64 while G was obtained by a five-step reaction (for details of the synthesis and characterization of G, see the Supporting Information). 1H NMR spectroscopy studies were carried out to investigate the binding properties between WP5 and guest G, and it was found that when 2.0 equiv. of WP5 was added to the solution of G (since G contains two trimethylammonium groups in both terminals, which could efficiently bond with WP5),67 the signals of Hb, Hc, Hd, He, Hf, and Hh from G shifted upfield remarkably because of the complexation induced shielding effects. Moreover, the signals of H1, H2, H3 from WP5 and Hp of G exhibited slight downfield chemical shift changes probably because the complexation


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between WP5 and G led to the decrease of the electron density for these protons (Figure 1). The above results revealed that the positively charged trimethylammonium head of G was threaded into the cavity of the WP5 host, resulting in the formation of a pseudorotaxane structure.

Figure 1. 1H NMR (400 MHz, D2O, 298 K) spectra: (a) G (2.0 mM), (b) G (2.0 mM) and WP5 (4.0 mM), and (c) WP5 (4.0 mM). To quantitatively assess the stoichiometry for WP5 and G, as well as their exact binding stoichiometry, isothermal titration calorimetry (ITC) was performed in neutral conditions at 298 K. It was found that the titration result could not be fitted well by using the normal “one set of binding sites” model. Only when the calorimetric traces were fitted with a sequential binding model in which 2:1 host−guest complexes were considered, a good fit was obtained, and the representative titration curve was shown in Figure 2. Based on the obtained ITC data, the stoichiometry between WP5 and G was WP5/G = 2:1, and the association constant (Ka1 and Ka2) was estimated to be (2.55 ± 0.72) × 105 M-1 and (3.11 ± 0.79) × 105 M-1, respectively. As we all known, the π-π interactions, C−H…π interactions, hydrogen bond, and van der Waals contribute


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for a favorable enthalpy change, while the electrostatic and hydrophobic interactions would like to make the entropic changes. Thus, the multiple electrostatic and hydrophobic interactions, as well as the cooperative π-π interactions are the main driven forces for such host–guest complexation, which leads to form a stable 2:1 bola-type amphiphilic WP5⊃ ⊃G inclusion complex.

Figure 2. Microcalorimetric titrations of WP5 with G in water at 298 K. (a) Raw ITC data of microcalorimetric titrations for 25 sequential injections (1.2 µL per injection) of WP5 solution (0.25 mM) into the aqueous solution of G (0.01 mM); (b) The heat effects of the complexation between WP5 and G for each injection, obtained by subtracting the dilution heat from the reaction heat, which was fitted by computer simulation by using the “sequential binding sites” model.


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Construction of Supramolecular Vesicles Based on WP5⊃ ⊃G Complex. With above bolatype amphiphilic WP5⊃ ⊃G inclusion complex in hand, we further applied it to construct highordered aggregates in water. Before investigating the aggregation of WP5⊃ ⊃G, it is necessary to know the aggregation behavior of free G. Dynamic light scattering (DLS) measurement of G solution (1 × 10-4 M) showed no appreciable signal, confirming that free G does not tend to form large-sized aggregates under the measured concentration. However, with addition of WP5 to above G solution, a light opalescence and notable Tyndall effect could be clearly observed (Figure 3a), suggesting the existence of abundant nanoparticles in the measured solution. Moreover, upon gradually adding the WP5 solution to G, the intensity of opalescence notably changed. Therefore, we further determined the best molar ratio between WP5 and G for constructing supramolecular aggregates by optical transmittance tests, and the best molar ratio between WP5 and G for generating such aggregates was deemed about 8:1 ([G]/[WP5]) (for details, see Supporting Information, Figure S10). Based on the obtained best molar ratio, the critical aggregation concentration (CAC) for WP5⊃ ⊃G was determined to be 1.41 × 10-5 M (Figure S11, Supporting Information). And then, zeta-potential assays were performed to investigate the stability of the nanoparticle solutions, and the results showed a negative ζpotential (−12.23 mV) for the formed WP5⊃ ⊃G vesicular solution at the best molar fraction. Taking into consideration of the repulsive-force-induced increasing-stability of the nanoparticles, the molar ratio of [G]/[WP5] = 5:1 (ζ-potential = −30.27 mV) was used for further investigating the stimuli-responsive behavior of the obtained nanoparticles and their potential applications in drug delivery. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) were employed to further investigate the size distribution and morphology of the self-assembled


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nanostructures formed by the bola-type WP5⊃ ⊃G amphiphile. As is shown in Figure 3b, WP5⊃ ⊃G assemblies exhibited a narrow size distribution with an average diameter of 184.5 nm. Meanwhile, TEM images clearly illustrated the formation of a number of spherical structures with diameters ranging from 140 to 170 nm. And the contrast of periphery and central parts of the spherical structures was easily distinguishable, which indicated the formation of hollow vesicular structures. Additionally, from the TEM images, the thickness of the vesicle membrane was measured to be about 5 nm, which fit approximately with the extended length of a single molecule estimated by 3D molecular structural modeling of WP5⊃ ⊃G complex in a 2:1 molar ratio (Figure S13, Supporting Information). Therefore, we deduced that the formed WP5⊃ ⊃G vesicles possess a monolayer structure as shown in Scheme 1, where both interior and exterior surfaces of the vesicles were the hydrophilic carboxylate groups of WP5, whereas, the core layer was composed by the hydrophobic aliphatic chains and naphthalimide groups of G.

Figure 3. (a) Tyndall effect of WP5⊃ ⊃G (left) and G (right); (b) DLS data of the WP5⊃ ⊃G aggregates; TEM images: (c) WP5⊃ ⊃G aggregates; (d) enlarged image of c.


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Stimuli-Responsiveness of the WP5⊃ ⊃G Vesicles. Stimuli-responsive properties are often incorporated into supramolecular nanocarrier systems, which would be liable to overcome certain systemic and intracellular delivery barriers. As we all known, the carboxylate groups on WP5 will be protonated under acidic condition and the generated pillar[5]arene in its acid form is not soluble in water, which will be precipitated from the acidic aqueous solution. Moreover, the disulfide linkages on G can be cleaved by excess glutathione (GSH), result in the complete destruction of the original structure, and the generated fragments cannot form stable nanoparticles any more. Therefore, these bola-type supramolecular vesicles will possess both pH- and GSH-responsiveness. As expected, the Tyndall effect disappeared after adding GSH (10 mM) to the WP5⊃ ⊃G solution, and accompanied by the generation of large precipitates; simultaneously, no vesicle structures could be detected from the TEM image (Figure S13, Supporting Information). Furthermore, when adjusting the solution pH to 5.0, we could also find the similar phenomenon. All of results proved that the above-mentioned bola-type supramolecular vesicles had excellent pH- and GSH-responsiveness, which enable them to load cargos at physiological condition and release them in acidic environment and/or reduction stimulation. Drug Loading and Controllable Release Behavior. As an ideal nanocarrier for drug delivery, it should have good stability in normal tissues but achieve high accumulation in tumor location for efficient passive targeting, and then show responsiveness to the tumor internal or external stimuli to release the loaded drugs. According to the dual-responsive character of WP5⊃ ⊃G vesicles, herein, hydrophobic anticancer drug DOX (doxorubicin) was selected to study the drug loading efficiency and release behavior of the obtained drug-loaded vesicles. Subsequently, DOX was successfully encapsulated into the hydrophobic Stern layer of the


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WP5⊃ ⊃G vesicles by passive loading. In comparison with the blank vesicular solution, the solution of DOX-loaded vesicles turned from colorless to rosiness; furthermore, fluorescence intensity of drug-loaded vesicles became much stronger in characteristic emission range of DOX from 550 to 650 nm, after being treated with Triton X-100 to achieve the complete release of DOX (Figure 4b). Moreover, it was found that DOX-loaded vesicles were larger in size than the blank vesicles based on the DLS results (Figure 4a). And DOX-loaded vesicles turned to be ellipse shape as evidenced by TEM images (Figure 4c and 4d). The above results suggested that DOX was successfully encapsulated into the WP5⊃ ⊃G vesicles. We further measured the drugloading efficiency of DOX-loaded vesicles by fluorescence spectra, and it was confirmed that the maximal drug-loading efficiency could be attained up to 13.7 %, revealing that this supramolecular vesicles have good loading ability for the hydrophobic drugs.

Figure 4. (a) DLS data of the DOX-loaded vesicles; (b) Fluorescence spectra of DOX-loaded vesicles, and DOX-loaded vesicles after being treated with Triton X-100; (c) TEM images of DOX-loaded vesicles; (d) TEM images of DOX-loaded vesicles staining by uranyl acetate.


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Next, the drug release behavior of DOX-loaded vesicles was further investigated under GSH and/or in acidic environment. As exhibited in Figure 5a, almost no obvious leakage of DOX could be detected under both GSH-free and neutral conditions, suggesting that the DOXloaded vesicles were almost stable under normal physiological condition. However, upon gradually adding GSH to the DOX-loaded vesicular solutions, GSH-induced DOX release could be observed clearly. As shown in Figure 5a, increasing the GSH concentration from 2 to 10 mM resulted in a gradual enhancement of cumulative drug release percentage from 52.1% to 81.5% within 60 min. Furthermore, when acidic and reduction environment were synchronously exerted on the DOX-loaded vesicles, the cumulative release of DOX was significantly accelerated under the co-triggered conditions (Figure 5b). In comparison with normal cells, the cancer cells have acidic microenvironment with the presence of much higher GSH concentration (1-11 mM).68 Therefore, dual acid/GSH-promoted rapid drug release is a very helpful feature in cancer therapy.69 In addition, the good stability of DOX-loaded vesicles under physiological condition could obviously reduce the toxicity and side effects of DOX for normal tissues, which is specifically significant for developing efficient drug delivery systems.


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Figure 5. The time-dependent drug release of DOX-loaded vesicles under different conditions: (a) with different concentrations of GSH at pH = 7.4, (b) with the presence of 5 mM GSH under different pH conditions. Cytocompatibility and Anticancer Efficiency Assay. As a drug carrier, good cytocompatibility of the vector was essential for biomedical and clinical applications. Therefore, the evaluation of cytocompatibility for both host (WP5) and guest (G) molecules at different concentrations against MRC-5 cells (normal human fetal lung fibroblast cell line) was carried out by MTT assay. In Figure 6, the minimal influence on cell viability and proliferation for MRC-5 cells incubated with WP5 was obtained with the concentrations ranging from 5 to 80 µM. With respect to G, although the cell viability was gradually reduced with increasing analyte concentration, the cell viability was still above 60% when the concentration of G was increased to 80 µM. The cytotoxicity of G especially at high concentration may be caused by its higher positive charge. All of the above results indicated the good biocompatibility for this bola-type supramolecular drug nanocarrier in the detected low concentration region.

Figure 6. In vitro biocompatibility of WP5 and G against MRC-5 cells after 24 h incubation (p < 0.05).


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Next, the anticancer efficiency of the drug nanocarriers was further evaluated, where A549 (human alveolar adenocarcinoma cell line) and HeLa cells (human cervical carcinoma cell line) were incubated with blank vesicles, free DOX·HCl, and DOX-loaded vesicles for 24 h, respectively. It was found that the relative cancer cell viability in the blank vesicle groups were much higher than that in the DOX-loaded vesicle groups, and the DOX-loaded vesicle groups showed much better therapeutic effects than free DOX·HCl groups at a wide range of concentrations (Figure 7 and Figure S14). Such phenomenon can be attributed to the following reasons: first, the efficient accumulation of DOX-loaded vesicles in cancer cells due to EPR effect; second, as shown in Figure S15, naphthalene imide-containing guest G could strongly interact with DNA, which caused the obviously inhibition of the proliferation ability of cancer cells.70 Moreover, it is noteworthy that A549 cells showed lower relative viabilities than HeLa cells when treated with DOX-loaded vesicles under the same conditions. The main reason might be that DOX has a much stronger interaction with A549 cells than HeLa cells, leading to the dramatically cell death for A549 cells. The above results indicated that loading the free drugs to the supramolecular nanocarriers could dramatically increase its anticancer efficiency but remarkably reduce its side effects to normal cells. Therefore, this kind of WP5⊃ ⊃G supramolecular vesicles based on the noncovalent host−guest interactions is a promising drug delivery system.


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Figure 7. In vitro cytotoxicities of DOX-loaded vesicles and DOX·HCl against (a) A549 cells and (b) HeLa cells, respectively, after 24 h incubation (p < 0.05). In vitro Cellular Uptake and Subcellullar Localization. The celluar uptaking and intracellular drug releasing behaviors of DOX-loaded vesicles were further evaluated by the confocal laser scanning microscopy (CLSM) and flow cytometry assay toward HeLa cells. Since the cellular internalization of free DOX·HCl has been investigated in our previous work,63 herein we mainly focused on the cellular internalization of DOX-loaded vesicles. Hochest and LysoTracker Red were used to label the nucleus and lysosomes, respectively (Hochest was shown in blue and LysoTracker was shown in green) for colocalizing these nanocarriers. Initially, HeLa cells were incubated with the same concentration of DOX-loaded vesicles at 37°C for 1 h and 4 h, respectively. In Figure 8, the red fluorescence of DOX appeared in cells after incubation for 1 h, demonstrating that DOX-loaded vesicles were successfully internalized by cancer cells. When the incubation time was increased to 4 h, HeLa cells showed strong intracellular red fluorescence. As we known, the fluorescence intensity observed in cells treated with free drugs or drug-loaded nanoparticles can be considered to be consistent with the concentration of the drug that internalized into the cells.66 Therefore, the intracellular fluorescence can clearly demonstrate the increasing concentration of DOX internalized into the


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cells with extended incubation time. Moreover, after incubation for 4 h, the lysosomes of HeLa cells displayed intense yellow fluorescence, which revealing the co-localization of lysosomes with DOX-loaded vesicles. The above results implied that these DOX-loaded vesicles entered into cancer cells through endocytosis and could localize in lysosomes, where the microenvironment with low-pH and high GSH concentration lead to the dissociation of the DOX-loaded vesicles and then the encapsulated DOX molecules were released into the cytosol, and finally entering the nucleus for cancer treatment.

Figure 8. Cellular uptake and subcellular localization of DOX-loaded vesicles in HeLa cells examined by CLSM after incubating at 37°C for 1 h and 4 h, respectively. (Scale bars: 20 µm). Subsequently, the quantitative experiments performed by flow cytometry, which further revealed that the DOX-loaded vesicles have much higher cellular uptake activity than free DOX·HCl. As shown in Figure 9, HeLa cells without any treatment were selected as a control, which displayed a negligible level of auto-fluorescence. However, the fluorescence intensity of DOX-loaded vesicles groups always exceed the free DOX·HCl groups after incubating with the


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same concentration of DOX both for 1 h and 4 h, respectively. Combining all above qualitative and quantitative results, we could conclude that these smart supramolecular nanocarriers could efficiently delivery anticancer drugs into cancer cells for the treatment of malignant tumor.

Figure 9. Flow cytometric profiles of HeLa cells after incubating with DOX·HCl or DOXloaded vesicles at 37°C for 1 h and 4 h, respectively. Conclusion In summary, we have successfully constructed an intelligent and dual stimuli-responsive drug delivery system by employing a bola-type supramolecular amphiphile WP5⊃G as the building unit. The resulting WP5⊃G amphiphile based on host-guest interaction could selfassemble into supramolecular vesicles in water, which could effectively encapsulate DOX to give DOX-loaded vesicles. And the loaded drug could be rapidly released at acidic microenvironment with high GSH concentration, allowing for the controlled drug release. Furthermore, cytotoxicity experiments confirmed that DOX-loaded vesicles could dramatically enhance the anticancer efficiency of free DOX for cancer cells but also effectively reduce its side effects to normal cells based on their pH/GSH dual-responsiveness. Cellular uptake and subcellular localization experiments proved that these supramolecular vesicles entered cancer


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cells mainly by endocytosis and could accelerate the remarkable drug accumulation in tumor cells. The present study paves an alternative way to develop bola-type smart supramolecular nanovehicles with multistimuli-responsiveness, which have great potential applications in the fields of controlled release and drug delivery. Experimental Section For the synthesis procedures of the NDI guest G, see the Supporting Information. The experimental procedures for the fabrication of the supramolecular vesicles and the biological related experiments were performed in accordance with the previous work in our lab, 58, 59, 61 but with some modifications (see Supporting Information for details). ASSOCIATED CONTENT The Supporting Information is available free of charge on the ACS Publications website via the Internet http://pubs.acs.org. General information and experimental procedure, detailed synthesis procedure and characterization, best molar ratio, CAC, TEM, in vitro cell assay (PDF) AUTHOR INFORMATION Corresponding Author *Email: [email protected] (XH). *Email: [email protected] (DZ). Author Contributions ∥

Xin Liu and ∥Keke Jia contributed equally

Notes


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The authors declare no competing financial interest. ACKNOWLEDGMENT This work was supported by the National Basic Research Program of China (2014CB846004), National Natural Science Foundation of China (No. 21572101, 21472089), and Jiangsu Provincial Natural Science Foundation of China (BK20140595). We also thank Dr. Xichun Liu for his kind help in ITC experiments. REFERENCES (1)

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Graphical Abstract


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Dual-Responsive Bola-Type Supra-Amphiphile ... - ACS Publications

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