Proton Sponge Research Articles

The metal-mediated coupling of the diselenaundecaborate anion [Se2B9H10

The diselenaundecaborate anion, [Se2B9H10]− (1), is isolated in 49 % yield from the one-pot reaction of nido-B10H14, Proton Sponge and selenium in thf. In solution, 1 exhibits a fluxional interchange between symmetry-equivalent positions of its two selenium atoms and open-face ‘bridging’ hydrogen atom that cannot be frozen out on the NMR timescale. This is however resolved by solid-state boron-11 NMR spectroscopy, which shows the spectrum expected from a DFT/GIAO calculation. The treatment of two equivalents of 1 with HgCl2 in dichloromethane affords the conjoined binary cluster anion [(Se2B9H10)(SeB9H11)]− (2), in which the ‘bridging’ selenium atom in 1 links to an {arachno-SeB9H11} moiety. This metal-mediated linking of selenaborane clusters potentially opens a gateway to new macropolyhedral heteroborane assemblies.

Organometallic Synthesis of 2,3,6,7-Tetrasubstituted 1,8-Bis(dimethylamino)naphthalenes for Investigation of the Double Buttressing Effect in Proton Sponges.

The lithiation of 2,7-disubstituted derivatives of 1,8-bis(dimethylamino)naphthalene (DMAN, proton sponge) bearing potentially ortho-directing OMe, NMe2, and SMe groups was studied. It has been shown that OMe groups facilitate selective dual β-lithiation of the naphthalene moiety while the 2(7)-NMe2 groups allow only monolithiation presumably due to the decreased acidity of the ring C-H bonds and conformational immobilization after coordination to the lithium atom. In contrast, the SMe groups provided no ring lithiation and underwent deprotonation of their methyl fragment. The first representatives of previously unknown 2,3,6,7-tetrasubstituted DMANs have been synthesized in good yield after treatment of 2,7-dimethoxy-3,6-dilithio DMAN with the appropriate electrophiles (MeI, Me2S2, Me3SiCl, DMF, etc.). Because the exceedingly high basicity of 2,7-dimethoxy DMAN is commonly attributed to the so-called "buttressing effect" (BE), the availability of 2,3,6,7-tetrasubstituted species provided the first opportunity to study the double BE version. Using X-ray diffraction and basicity measurements, we showed that due to the high conformational mobility of the methoxy groups, the most striking manifestations of double BE are the strong planarization of peri-NMe2 groups and a significant decrease in basicity, while the length and the other properties of the intramolecular NHN hydrogen bond in the corresponding protonated species undergo minor changes.

Stimuli-responsive liposomal nanoformulations in cancer therapy: Pre-clinical & clinical approaches.

The site-specific delivery of antitumor agents is of importance for providing effective cancer suppression. Poor bioavailability of anticancer compounds and the presence of biological barriers prevent their accumulation in tumor sites. These obstacles can be overcome using liposomal nanostructures. The challenges in cancer chemotherapy and stimuli-responsive nanocarriers are first described in the current review. Then, stimuli-responsive liposomes including pH-, redox-, enzyme-, light-, thermo- and magneto-sensitive nanoparticles are discussed and their potential for delivery of anticancer drugs is emphasized. The pH- or redox-sensitive liposomes are based on internal stimulus and release drug in response to a mildly acidic pH and GSH, respectively. The pH-sensitive liposomes can mediate endosomal escape via proton sponge. The multifunctional liposomes responsive to both redox and pH have more capacity in drug release at tumor site compared to pH- or redox-sensitive alone. The magnetic field and NIR irradiation can be exploited for external stimulation of liposomes. The light-responsive liposomes release drugs when they are exposed to irradiation; thermosensitive-liposomes release drugs at a temperature of >40°C when there is hyperthermia; magneto-responsive liposomes release drugs in presence of magnetic field. These smart nanoliposomes also mediate co-delivery of drugs and genes in synergistic cancer therapy. Due to lack of long-term toxicity of liposomes, they can be utilized in near future for treatment of cancer patients.

Resolving subcellular pH with a quantitative fluorescent lifetime biosensor

Changes in sub-cellular pH play a key role in metabolism, membrane transport, and triggering cargo release from therapeutic delivery systems. Most methods to measure pH rely on intensity changes of pH sensitive fluorophores, however, these measurements are hampered by high uncertainty in the inferred pH and the need for multiple fluorophores. To address this, here we combine pH dependant fluorescent lifetime imaging microscopy (pHLIM) with deep learning to accurately quantify sub-cellular pH in individual vesicles. We engineer the pH sensitive protein mApple to localise in the cytosol, endosomes, and lysosomes, and demonstrate that pHLIM can rapidly detect pH changes induced by drugs such as bafilomycin A1 and chloroquine. We also demonstrate that polyethylenimine (a common transfection reagent) does not exhibit a proton sponge effect and had no measurable impact on the pH of endocytic vesicles. pHLIM is a simple and quantitative method that will help to understand drug action and disease progression.

Chemical Modulation of Glucose Metabolism with a Fluorinated CaCO3 Nanoregulator Can Potentiate Radiotherapy by Programming Antitumor Immunity.

Tumor hypoxia and acidity are well-known features in solid tumors that cause immunosuppression and therapeutic resistance. Herein, we rationally synthesized a multifunctional fluorinated calcium carbonate (fCaCO3) nanoregulator by coating CaCO3 nanoparticles with dopamine-grafted perfluorosebacic acid (DA2-PFSEA) and ferric ions by utilizing their coordination interaction. After PEGylation, the obtained fCaCO3-PEG showed high loading efficacy to perfluoro-15-crown-5-ether (PFCE), a type of perfluorocarbon with high oxygen solubility, thereby working as both oxygen nanoshuttles and proton sponges to reverse tumor hypoxia and acidity-induced resistance to radiotherapy. The as-prepared PFCE@fCaCO3-PEG could not only function as long-circulating oxygen nanoshuttles to attenuate tumor hypoxia but also neutralize the acidic tumor microenvironment by restricting the production of lactic acid and reacting with extracellular protons. As a result, treatment with PFCE@fCaCO3-PEG could improve the therapeutic outcome of radiotherapy toward two murine tumors with distinct immunogenicity. The PFCE@fCaCO3-PEG-assisted radiotherapy could also collectively inhibit the growth of unirradiated tumors and reject rechallenged tumors by synergistically eliciting protective antitumor immunity. Therefore, our work presents the preparation of fluorinated CaCO3 nanoregulators to reverse tumor immunosuppression and potentiate radiotherapy through chemically modulating tumor hypoxic and acidic microenvironments tightly associated with tumor glucose metabolism.

Effect of molecular weight of polysaccharide on efficient plasmid DNA delivery by polyethylenimine‐polysaccharide‐Fe(III) complexes

AbstractPolyethylenimine (PEI) is considered as the gold standard for evaluating non‐viral gene vectors due to its proton sponge effect and high transfection efficiency. However, its cytotoxicity limits its application. Rice bran polysaccharide (RBP) is rich in nature and beneficial for preparation of biodegradable biomaterials. In this work, three PEI modified RBP‐Fe complexes (PIP‐1, PIP‐2, and PIP‐3) were prepared based on RBP with different molecular weight (Mw). Then the potential of PIP complexes for plasmid DNA (pDNA) delivery and the influence of Mw was studied. PIP complexes showed better safety and higher gene transfection efficiency than PEI. PIP‐1 was prepared by RBP with lower Mw and could formed PIP‐1/pDNA nanoparticle with lower particle size which exhibited higher gene transfection efficiency than other PIP complexes prepared by RBP with higher Mw. The results indicate that the PIP/pDNA complex exhibits a multi‐pathway cellular uptake mechanism. Clathrin dependent and caveolin dependent endocytosis and macropinocytosis were involved in the cellular uptake process of PIP/pDNA complex. It is hoped that this study can provide useful enlightenment for the research and development of gene vector based on polysaccharide.

Self-fluorescence property of octa-arginine functionalized hydroxyapatite nanoparticles aids in studying their intracellular fate in R1 ESCs

Deciphering the endocytosis mechanisms of nanoparticle entry in cells is crucial to understand the fate of nanoparticles and the biological activity of the transported cargo. Such studies require the use of reporter agents such as fluorescent markers. Previously, we have reported the synthesis of self-fluorescent HAp nanoparticles as efficient nucleic acid delivery agents in prokaryotic and eukaryotic cells. Here, we show the application of biocompatible self-fluorescent nano delivery vehicle based on HAp and CPP- octa-arginine as an efficient system to study the mechanisms of intracellular fate of a gene delivery agent. The pathway of octa-arginine functionalized HAp NP (R8HNP) and HAp NP uptake in R1 ESCs was elucidated using confocal microscopy with the help of endocytic inhibitors. The NPs mainly enter R1 ESCs by clathrin mediated and macropinocytosis pathways. It was established that the NPs escape endosomal degradation by proton sponge effect owing to their ability to buffer the pH and trigger osmotic rupture. The functionalization of CPP, effectively enhanced the internalization and endosomal escape in R1 ESCs. The detailed results of these studies are outlined in this manuscript.

Surface functionalization of lipidic core nanoparticles with albumin: A great opportunity for quinacrine in lung cancer therapy

Bovine serum albumin (BSA) decorated lipidic core nanoparticles were constructed for the delivery of quinacrine (Quin) to enhance docetaxel (Dtx) dose-dependent behavior against A-549 lung cancer cells through the inhibition of the Nrf2 signal pathway and arresting the Sub-G1 cell cycle mechanism. Chitosan was chosen as a suitable stabilizer in addressing the proton sponge effect with BSA providing the favorable protein corona formation along with low toxicity. The optimized formulation was characterized in terms of particle size, zeta potential, morphology, polydispersity index and encapsulation efficiency (EE). The mechanism of cytotoxicity, apoptosis and cell cycle arrest along with the determination of the combined efficacy of Quin with Dtx were investigated by MTT assay, flow cytometric assay, CompuSyn and HSA synergy score analysis. Real-time PCR and Western blot analysis were performed to quantify Nrf2-dependent genes and protein levels, respectively. The results demonstrated an average particle size of 112 ± 16.2 nm, a positive surface charge of 15 ± 0.1 mV along with an EE of 65.4 ± 2.6%. Quin NPs reduced the proliferation of lung cancer cells from 68 ± 6.1% to 49 ± 4.5% (p < 0.05) along with a two-fold surge in the percentage of apoptosis (p < 0.05). The incubation of A549 lung cancer cells with Quin NPs caused a significant decrease in Nrf2, NQO1 and MRP1 associated with an increase in BAD mRNA levels compared to the other groups (p < 0.05). Combinational treatment of both Quin + Dtx, as well as Quin NPs + Dtx, induced a remarkable G2M arrest and sub-G1 apoptosis. The results, therefore, indicate that Quin-loaded nanoparticles have the ability to enhance the anti-proliferation efficacy of Dtx against A549 lung cancer by inhibiting Nrf2, increasing synergistic interaction with lower doses of Dtx and arresting cell cycle in the Sub-G1 thus leading to the induction of apoptosis.

Nanocapping-enabled charge reversal generates cell-enterable endosomal-escapable bacteriophages for intracellular pathogen inhibition.

Bacteriophages (phages) are widely explored as antimicrobials for treating infectious diseases due to their specificity and potency to infect and inhibit host bacteria. However, the application of phages to inhibit intracellular pathogens has been greatly restricted by inadequacy in cell entry and endosomal escape. Here, we describe the use of cationic polymers to selectively cap negatively charged phage head rather than positively charged tail by electrostatic interaction, resulting in charge-reversed phages with uninfluenced vitality. Given the positive surface charge and proton sponge effect of the nanocapping, capped phages are able to enter intestinal epithelial cells and subsequently escape from endosomes to lyse harbored pathogens. In a murine model of intestinal infection, oral ingestion of capped phages significantly reduces the translocation of pathogens to major organs, showing a remarkable inhibition efficacy. Our work proposes that simple synthetic nanocapping can manipulate phage bioactivity, offering a facile platform for preparing next-generation antimicrobials.

Non-viral CRISPR activation system targeting VEGF-A and TGF-β1 for enhanced osteogenesis of pre-osteoblasts implanted with dual-crosslinked hydrogel.

Healing of large calvarial bone defects remains challenge but may be improved by stimulating bone regeneration of implanted cells. The aim of this study is to specially co-activate transforming growth factor β1 (TGF-β1) and vascular endothelial growth factor (VEGF-A) genes expressions in pre-osteoblast MC3T3-E1 cells through the non-viral CRISPR activation (CRISPRa) system to promote osteogenesis. A cationic copolymer carrying nucleus localizing peptides and proton sponge groups dimethyl-histidine was synthesized to deliver CRISPRa system into MC3T3-E1 cells with high cellular uptake, lysosomal escape, and nuclear translocation, which activated VEGF-A and TGF-β1 genes expressions and thereby additively or synergistically induced several osteogenic genes expressions. A tunable dual-crosslinked hydrogel was developed to implant the above engineered cells into mice calvaria bone defect site to promote bone healing in vivo. The combination of multi-genes activation through non-viral CRISPRa system and tunable dual-crosslinked hydrogel provides a versatile strategy for promoting bone healing with synergistic effect.

Gold Nanocluster-Encapsulated Hyperbranched Polyethyleneimine for Selective and Ratiometric Dopamine Analyses by Enhanced Self-Polymerization.

The exploitation of selective and sensitive dopamine (DA) sensors is essential to more deeply understand its biological function and diagnosis of related diseases. In this study, gold nanocluster-encapsulated hyperbranched polyethyleneimine (hPEI-Au NCs) has been explored as the specific and ratiometric DA nanoprobe through hPEI-assisted DA self-polymerization reactions. The Au NCs encapsulation not only provides a fluorescent internal reference but also enhances the DA self-polymerization by weakening the proton sponge effect of the hPEI layer. Rapid and sensitive DA detection is realized through the proposed hPEI-Au NC nanoprobe with a limit of detection of 10 nM. The favorable selectivity over other possible interferents including amino acids, sugars, and salts is due to the specific self-polymerization reaction. The DA analysis in urine samples with small relative standard deviations has been accomplished with an hPEI-Au NC nanoprobe.

Reverse relation between cytotoxicity and Polyethylenimine/DNA ratio, the effect of using HEPES-buffered saline (HBS) medium in gene delivery

Polyethyleneimine (PEI) is considered a promising cationic polymer in non-viral gene delivery. DNA binding properties and other biochemical characteristics of PEI such as the proton sponge phenomenon, offered the branched 25 kDa PEI to be widely used for therapeutic DNA delivery, although the possible cytotoxic effects and the best conditions of PEI preparation are not still well recognized. While higher PEI/Plasmid ratios have increased transfection efficiencies, it induces more cell stress and toxicity. Considering that the PEI particle size and resulting cytotoxicity are affected by media ions, we used Neuro2A cells to assess the cell stress properties of PEI/Plasmid complexes prepared in a HEPES-buffered saline medium. Delivery of a plasmid containing EGFP happened in all increasing ratios of PEI/plasmid from 0.5, 2, 4, and 6, while higher ratios induced less unfolded protein response as evidenced by lower transcription of ER stress markers Grp78, Atf4, Chop, Xbp1, and induced Xbp1 splicing. These data were also supported by MTT cytotoxicity assay results. These findings indicate that preparing higher PEI/plasmid ratio complexes (using the equivalent of 200 ng DNA) in the HBS medium leads to less cytotoxicity.

“Proton sponge” effect and apoptotic cell death mechanism of Agx-Re6 nanocrystallites derived from the assembly of [{Re6S8}(OH)6–n(H2O)n]n–4 with Ag+ ions

The present work introduces heterometallic Agx-Re6 (Re6 = [{Re6S8}(OH)6–n(H2O)n]n–4) nanocrystalline colloids self-generated under the mixing of Re6 with [Ag2L2](BF4)2 (L = pyridine-2-ylphospholane) in aqueous solutions. The easy protonation/deprotonation of the Re6 units incorporated into the Agx-Re6 colloids provides their negative surface charging and low aggregation in neutral buffered solutions and the recharging of the colloids in weak acidic solutions modeling lysosomal environment. The surface-exposed Ag(I) ions are prerequisite for binding of glutathione (GSH), which is visualized through the red emission of the Re6 units. The treating of Agx-Re6 by polyethylenimine (PEI) both recharges the colloids and restricts their binding with GSH. Quantitative colocalization analysis of the red emitting Agx-Re6 colloids allows to visualize the low lysosomal trafficking of both initial and PEI-treated colloids, which indicates that pH-driven protonation of either Re6 or amino-groups of PEI provides the similar “proton sponge” effects. The apoptotic assay results indicate high contribution of the apoptotic cell death mechanism for both [Ag2L2](BF4)2 and Agx-Re6 colloids, while their PEI-coating changes the cell death mechanism to the non-apoptotic one and enhances the cytotoxicity.

Dual-enzyme catalytic nanosystem-mediated ATP depletion strategy for tumor elimination via excessive autophagy pathway

Excessive autophagy is a promising pathway to eliminate tumor cells, which causes excessive damages to overload the degradation capacity of lysosomes. Designing a nanosystem that can compromise the function of lysosomes and induce severe damages simultaneously is essential to activate excessive autophagy, but is still rarely explored. Herein, a dual-enzyme catalytic nanosystem with adenosine triphosphate (ATP) depletion capacity was developed for tumor elimination via the excessive autophagy pathway. This nanosystem consisted of polyvinyl pyrrolidone-stabilized polyaniline (PANI-PVP) core, Pt nanoparticles (NPs) decoration, and glucose oxidase (GOx) payload. After being internalized by tumor cells, this formed PANI-Pt-GOx-PVP NPs could escape from the lysosome through the proton sponge effect, causing lysosomal swelling and rupturing. The Pt-GOx dual-enzyme catalytic nanosystem consumes glucose to cause severe ATP shortage. This energy depletion microenvironment can not only generate intracellular damage during the GOx-mediated starvation process, but also induce mitophagy through AMPK activation. The increased accumulation of damage causes excessive autophagy, overloading the degradation capacity of lysosomes. In vivo data demonstrated that tumor growth tendency was significantly suppressed through this ATP depletion-induced excessive autophagy strategy.

Dual Inhibitions on Glucose/Glutamine Metabolisms for Nontoxic Pancreatic Cancer Therapy.

Glucose and glutamine are two principal nutrients in mammalian cells that provide energy and biomass for cell growth and proliferation. Especially in cancer cells, glutamine could be a main alternative for energy and biomass supply once glucose metabolism is suppressed. Therefore, single inhibition of enzymes in either glucose metabolism or glutaminolysis, though maybe efficient in vitro, is far from being satisfactory for efficient in vivo cancer therapy. Here, we proposed a new strategy for dual inhibitions on both glucose and glutamine metabolisms concurrently by silencing mutated gene Kras and glutaminase 1 (GLS1) via nanomaterial-based siKras and siGLS1 delivery, rather than conventional highly toxic chemodrugs. Such a combination therapy could overcome the challenge that glucose and glutamine are alternatives to each other in the biosynthesis and energy production for cancer cells, resulting in much elevated treatment efficacy. In addition, layered double hydroxide (LDH), the siRNA carrier, enables an enhanced gene delivery efficiency compared to the commercial transfection agent Lipofectamine 2000. Briefly, Mg-Al LDH nanosheets, loaded with siKras and siGLS1 onto their surfaces by electrostatic adsorption, could release siRNA from lysosomes into the cytoplasm via the proton sponge effect of LDH, favoring the siRNA stability and gene silencing efficiency enhancements. The thus released siRNA could downregulate the expressions of Kras, GLS1, and other enzymes involved in glucose metabolism, resulting in the downregulations of ATP and other metabolites. Such a biosafe LDH/siRNA nanomedicine is able to efficiently suppress the growth of xenografts through cancer cell proliferation suppression, displaying its great potential as a simultaneous glucose/glutamine metabolism coinhibitor for treating pancreatic cancer.

The structural versatility of proton sponge bismuth halides

Hybrid halometalates containing lead, tin, bismuth and antimony and organic cations have recently shown a bevy of interesting photophysical properties. Aiming at finding chemically stable and thermally inert species, three halobismutate species of this class, crystallized with proton sponge-derived cations (PRSH), have been isolated as microcrystalline powders by mixing 1,8-bis(dimethylamino)-naphthalene (proton sponge, or PRS) and bismuth oxide in concentrated HX acids (X ​= ​Cl, Br and I). The two isomorphous (PRSH)3Bi2X9 (X ​= ​Br, I) species, containing isolated [Bi2X9]3- anions, are triclinic at room temperature and convert upon heating into a monoclinic structure through a displacive phase transformation, fully reversible for X ​= ​I and only partially for X ​= ​Br. At variance, (PRSH)BiCl4 is polymeric, and contains extended zigzagging 1D chains formed by edge-sharing BiCl6 octahedra. These species were extensively studied by synchrotron and laboratory X-ray powder diffraction measurements, which enabled to detect the evolution toward the two high-temperature β-phases (X ​= ​Br, I), to derive the structure of five different (significantly complex) species and to assess the thermal strain tensors in the different regimes. Additional thermal, spectroscopic and computational analyses completed the characterization of these materials.

Understanding the Biological Interactions of pH‐Swellable Nanoparticles

Inside Front Cover: Nanoparticles which undergo pH-induced swelling upon cell internalization are synthesized with tunable swelling and buffering capacity. Previous research has suggested such materials endosomal escape through both the proton sponge and pH-induced swelling mechanisms. However, in this work no endosomal escape was observed, suggesting limitations with these mechanisms. This is reported by Georgina K. Such and co-workers in article number 2100445.

Gold nanorods/tetrahedral DNA composites for chemo-photothermal therapy.

Combination therapy is extensively developed for cancer treatment in recent years due to its high efficiency. Herein, we constructed a nanocomposite based on gold nanorods (GNRs) and drug-loaded tetrahedral DNA nanostructures (TDN) for chemo-photothermal combinational therapy. Anti-tumor drug doxorubicin (DOX) was loaded via the insertion within GC base pairs of TDN. The aptamer AS1411 was attached to the apex of TDN (ATDN) to target tumor cells. The DOX-loaded DNA tetrahedron (ATDN-DOX) was compressed by the GNRs coated with PEI (GNRs@ATDN-DOX) to realize the photothermal function and lysosome escape. GNRs under the illumination of 808 nm infrared laser showed high photothermal conversion and stability due to the protection of PEI layer. The drug-loading capacity of ATDN-DOX was as high as 314 DOX molecules in per ATDN. The positive charge of PEI in GNRs@ATDN-DOX nanocomposites was utilized to achieve excellent cell penetration and induce proton sponge effect for lysosomal escape. The nanocomposites presented HeLa and 4T1 cells targeting and resulted in efficient anticancer activity.

Conjugation of Oligo-His Peptides to Magnetic γ-Fe2O3@SiO2 Core-Shell Nanoparticles Promotes Their Access to the Cytosol.

The endosomal entrapment of functional nanoparticles is a severe limitation to their use for biomedical applications. In the case of magnetic nanoparticles (MNPs), this entrapment leads to poor heating efficiency for magnetic hyperthermia and suppresses the possibility to manipulate them in the cytosol. Current strategies to limit their entrapment include functionalization with cell-penetrating peptides to promote translocation directly across the cell membrane or facilitate endosomal escape. However, these strategies suffer from the potential release of free peptides in the cell, and to the best of our knowledge, there is currently a lack of effective methods for the cytosolic delivery of MNPs after incubation with cells. Herein, we report the conjugation of fluorescently labeled cationic peptides to γ-Fe2O3@SiO2 core-shell nanoparticles by click chemistry to improve MNP access to the cytosol. We compare the effect of Arg9 and His4 peptides. On the one hand, Arg9 is a classical cell-penetrating peptide able to enter cells by direct translocation, and on the other hand, it has been demonstrated that sequences rich in histidine residues can promote endosomal escape, possibly by the proton sponge effect. The methodology developed here allows a high colocalization of the peptides and core-shell nanoparticles in cells and confirms that grafting peptides rich in histidine residues onto nanoparticles promotes NPs' access to the cytosol. Endosomal escape was confirmed by a calcein leakage assay and by ultrastructural analysis in transmission electron microscopy. No toxicity was observed for the peptide-nanoparticles conjugates. We also show that our conjugation strategy is compatible with the addition of multiple substrates and can thus be used for the delivery of cytoplasm-targeted therapeutics.

Enzyme-powered nanomotors with enhanced cell uptake and lysosomal escape for combined therapy of cancer

Low cellular uptake and lysosomal degradation are key difficulties of drug delivery in cancer treatment. Here, we report a novel nanomotor which can accomplish self-propulsion under the tumoral endogenous hydrogen peroxide to achieve high cell uptake rate and lysosomal escape. Calcium carbonate nanoparticles were employed as the core with polyethyleneimine (PEI) and catalase (CAT) acting as the shell to prepare the nanomotor via layer-by-layer self-assembly technology. To achieve local administration and slow release inside tumor, the nanomotors were loaded in Schiff-base hydrogel to build NM@hydrogel system. Taking advantage of the tumor microenvironment featured with an acidic pH and a high amount of hydrogen peroxide, the nanomotors were released from NM@hydrogel system in response to weak acidic tumor matrix. The released nanomotors can be autonomously propelled by the oxygen gradient generated from catalytic decomposition of hydrogen peroxide with a speed at 10.1±1.1 µm/s in 0.1 mM H2O2. Furtherly, combined with autonomous motion of nanomotors, specific tumor affinity strategy mediated by folic acid (FA) modification on the nanomotors was also proposed in the NM@hydrogel system, which the cell uptake rate was enhanced to about 80.6±2.0%. Furthermore, after internalization, the nanomotors could also efficiently escape from lysosomes owing to the proton sponge effect caused by PEI, CO2 produced by the degradation of CaCO3 nanoparticles and autonomous motion, which facilitated PTX and siRNA to reach their intracellular target, tubulin. PTX and siRNA strongly affected properties of tubulin and resulted in tumor cell apoptosis. In vitro and in vivo studies demonstrated excellent antitumor effect of the NM@hydrogel. Therefore, we anticipate that the proposed system would provide new insight in the drug delivery for cancer treatment.