Efficient Cellular Internalization Research Articles

A Novel Homodimer Peptide–Drug Conjugate Improves the Efficacy of HER2-Positive Breast Cancer Therapy

Tumor-targeting peptide-drug conjugates (PDCs) have become a focus of research in recent years. However, due to the instability of peptides and their short in vivo effective half-life, they have limited clinical application. Herein, we propose a new DOX PDC based on a homodimer HER-2-targeting peptide and acid-sensitive hydrazone bond, which could enhance the anti-tumor effect of DOX and reduce systemic toxicities. The PDC could accurately deliver DOX into HER2-positive SKBR-3 cells, with it showing 2.9 times higher cellular uptake than free DOX and enhanced cytotoxicity with respect to IC50 of 140 nM (vs. 410 nM for free DOX). In vitro assays showed that the PDC had high cellular internalization efficiency and cytotoxicity. In vivo anti-tumor experiments indicated that the PDC could significantly inhibit the growth of HER2-positive breast cancer xenografts in mice and reduce the side effects of DOX. In summary, we constructed a novel PDC molecule targeting HER2-positive tumors, which may overcome some deficiencies of DOX in breast cancer therapy.

Multifunctional biomimetic nanoplatform based on photodynamic therapy and DNA repair intervention for the synergistic treatment of breast cancer.

Photodynamic therapy (PDT) is a minimally invasive and locally effective treatment method, which has been used in the clinical treatment of a variety of superficial tumors. In recent years, PDT has received extensive attention due to its induction of immunogenic cell death (ICD). However, the repair mechanism of tumor cells and low immune response limit the further development of PDT. To this end, a multifunctional biomimetic nanoplatform 4T1Mem@PGA-Ce6/Ola (MPCO) is developed to co-deliver the photosensitizer Chlorin e6 (Ce6) and Olaparib (Ola) with the function of preventing DNA repair. The nanoplatform shows efficient tumor targeting and cellular internalization properties due to cell membrane camouflage, and Ce6 and Ola produce a significant synergistic anti-tumor effect under laser irradiation. Meanwhile, the nanoplatform can also activate the cyclic guanosine monophosphate-adenosine monophosphate synthase-interferon gene stimulator signaling (cGAS-STING) pathway to produce cytokines. The damage-associated molecular patterns induced by ICD can work with these cytokines to recruit and stimulate the maturation of dendritic cells and induce the systemic anti-tumor immune response. Overall, this multifunctional biomimetic nanoplatform integrating PDT, chemotherapy, and immunotherapy is highlighted here to boost anti-tumor therapy. STATEMENT OF SIGNIFICANCE: Self-repair of DNA damage is the most important reason for the failure of primary tumor eradication and the formation of secondary and metastatic tumors. To address this issue, a multifunctional biomimetic nanoplatform 4T1Mem@PGA-Ce6/Ola (MPCO) was developed to integrate a photosensitizer Chlorine a6 and a poly (ADP-ribose) polymerase inhibitor Olaparib. With tumor targeting ability and controlled release of drugs, the MPCO was expected to enhance tumor immunogenicity and facilitate antitumor immunity through the induction of immunogenic cell death as well as the activation of the cGAS-STING pathway. This study develops a promising combination strategy against tumors and has substantial implications for the prognosis of patients with breast cancer.

A dual-sensitive nanoparticle-mediated deepening synergistic therapy strategy involving DNA damage and ICD stimuli to treat triple-negative breast cancer.

Triple-negative breast cancer (TNBC) is one of the most aggressive cancers with an immunosuppressive microenvironment, and achieving a satisfactory effect from monotherapies, such as chemotherapy, photodynamic therapy (PDT) or immunotherapy, remains difficult. To solve this puzzle, a deepening synergistic therapy strategy of DNA damage and immunogenic cell death (ICD) stimuli was proposed. We engineered a doxorubicin (DOX) and 4-(hydroxymethyl) phenylboronic acid pinacol ester (PBAP) prodrug polymer, and encapsulated chlorin e6 (Ce6) to obtain the hyaluronidase (HAase) and H2O2 dual-sensitive responsive nanoparticles (Ce6/HDP NPs). The NPs displayed efficient intratumoral accumulation and cellular internalization properties due to the active targeting of the hyaluronic acid (HA). The dual DNA damage of the chemotherapy and ROS production directly caused tumor cell apoptosis. The strong ICD stimuli, which were induced by ROS production and GSH depletion, generated an amplified immunogenicity to activate tumor immunotherapy in vivo. In this manner, the NPs could significantly inhibit primary tumor, abscopal tumor, pulmonary metastasis and recurrent tumor in a subcutaneous 4T1 tumor model, with effective biosafety. This study has provided a promising deepening synergistic therapy strategy against TNBC.

Cell-Penetrating Peptide Conjugated Au Nanoclusters Selectively Suppress Refractory Lymphoma Cells via Targeting Both Canonical and Noncanonical NF-κB Signaling Pathways.

Activated B cell-like diffuse large B-cell lymphoma (ABC-DLBCL) is the most aggressive form of DLBCL, with a significantly inferior prognosis due to resistance to the standard R-CHOP immunochemotherapy. Survival of ABC-DLBCL cells addicted to the constitutive activations of both canonical and noncanonical NF-κB signaling makes them attractive therapeutic targets. However, a pharmaceutical approach simultaneously targeting the canonical and noncanonical NF-κB pathway in the ABC-DLBCL cell is still lacking. Peptide-conjugated gold nanoclusters (AuNCs) have emerged unique intrinsic biomedical activities and possess a great potential in cancer theranostics. Here, we demonstrated a Au25 nanocluster conjugated by cell-penetrating peptides that can selectively repress the growth of ABC-DLBCL cells by inducing efficient apoptosis, more efficiently than glutathione (GSH)-conjugated AuNCs. The mechanism study showed that the cell-penetrating peptides enhanced the cellular internalization efficiency of AuNCs, and the selective repression in ABC-DLBCL cells is due to the inhibition of inherent constitutive canonical and noncanonical NF-κB activities by AuNCs. Several NF-κB target genes involved in chemotherapy resistance in ABC-DLBCL cells, including anti-apoptotic Bcl-2 family members and DNA damage repair proteins, were effectively down-regulated by the AuNC. The emerged novel activity of AuNCs in targeting both arms of NF-κB signaling in ABC-DLBCL cells may provide a promising candidate and a new insight into the rational design of peptide-conjugated Au nanomedicine for molecular targeting treatment of refractory lymphomas.

Uncloaking cell-impermeant gold nanorods via tumor microenvironmental cathepsin B facilitates cancer cell penetration and potent radiosensitization

Major impediments to conveyance of intravenously administered drugs to tumors are biofouling, opsonization, and rapid clearance from the circulation by macrophages and reticuloendothelial phagocytes. Cloaking nanoparticles with stealth epilayers partly overcomes these hurdles but it also foils interactions with tumor cells. Here, we describe the synthesis, characterization, and validation of smart gold nanorods (GNRs) that spontaneously transform from inert passengers in the blood stream to active cell-penetrating nanoparticles within tumors to potently sensitize tumors to radiation therapy. Intrinsically cationic and cell-penetrating GNRs were shielded from phagocytosis with a cloaking polyethylene glycol epilayer containing an intervening cleavable peptide. In the absence of an external trigger, this epilayer is clipped off by the tumor microenvironmental protease, cathepsin B, in colorectal cancers to uncloak and expose the free-circulating native unPEGylated GNR that is readily internalized by cancer cells and turn into immovable small clusters of GNRs. Selective uncloaking of GNRs in the tumor reduced off-target toxicity confirmed by hematologic, biochemical, and histopathological analysis of blood, serum, and normal organs, respectively. Subsequent irradiation led to significant tumor growth delay and improved survival of mice. By addressing multiple barriers to efficient transport and cellular internalization of nanoparticles, our results demonstrate that clinically meaningful radiosensitization can be achieved with rationally designed GNRs.

Carrier‐free nanodrugs for stemness inhibition‐enhanced photodynamic therapy

AbstractCancer stem cell (CSC) has been considered a key driver of tumor growth, recurrence, and metastasis due to its self‐renewal and tumor initiation capacities called stemness. Stemness is also responsible for the high resistance of CSC to current therapeutic strategies, including photodynamic therapy (PDT) and chemotherapy. In this study, a carrier‐free nanodrug (designated as MKCe6 nanoparticle [NP]) self‐assembled by photosensitizer chlorin e6 (Ce6) and stemness inhibitor MK‐0752 was prepared for effective tumor repression. Stemness inhibition caused by MK‐0752 could sensitize CSCs to PDT. Benefiting from the high drug‐loading capacity and efficient cellular internalization, MKCe6 NPs exhibited good performance in PDT and stemness inhibition. In this way, effective tumor growth repression and tumorigenesis inhibition by MKCe6 NPs were observed both in vitro and in vivo. This self‐delivery nanodrug for stemness inhibition‐enhanced photodynamic therapy may provide new insights for clinical cancer therapy.

Development of WRAP5 Peptide Complexes for Targeted Drug/Gene Co-Delivery toward Glioblastoma Therapy.

Despite the great progress over the past few decades in both the diagnosis and treatment of a great variety of human cancers, glioblastoma remains the most lethal brain tumor. In recent years, cancer gene therapy focused on non-viral vectors which emerged as a promising approach to glioblastoma treatment. Transferrin (Tf) easily penetrates brain cells of the blood–brain barrier, and its receptor is highly expressed in this barrier and glioblastoma cells. Therefore, the development of delivery systems containing Tf appears as a reliable strategy to improve their brain cells targeting ability and cellular uptake. In this work, a cell-penetrating peptide (WRAP5), bearing a Tf-targeting sequence, has been exploited to condense tumor suppressor p53-encoding plasmid DNA (pDNA) for the development of nanocomplexes. To increase the functionality of developed nanocomplexes, the drug Temozolomide (TMZ) was also incorporated into the formulations. The physicochemical properties of peptide/pDNA complexes were revealed to be dependent on the nitrogen to phosphate groups ratio and can be optimized to promote efficient cellular internalization. A confocal microscopy study showed the capacity of developed complexes for efficient glioblastoma cell transfection and consequent pDNA delivery into the nucleus, where efficient gene expression took place, followed by p53 protein production. Of promise, these peptide/pDNA complexes induced a significant decrease in the viability of glioblastoma cells. The set of data reported significantly support further in vitro research to evaluate the therapeutic potential of developed complexes against glioblastoma.

Design of Cyclic Peptide-Based Nanospheres and the Delivery of siRNA.

In recent years, cyclic peptides have attracted much attention due to their chemical and enzymatic stability, low toxicity, and easy modification. In general, the self-assembled nanostructures of cyclic peptides tend to form nanotubes in a cyclic stacking manner through hydrogen bonding. However, studies exploring other assembly strategies are scarce. In this context, we proposed a new assembly strategy based on cyclic peptides with covalent self-assembly. Here, cyclic peptide-(DPDPDP) was rationally designed and used as a building block to construct new assemblies. With cyclo-(DP)3 as the structural unit and 2,2′-diamino-N-methyldiethylamine as the linker, positively charged nanospheres ((CP)6NS) based on cyclo-(DP)3 were successfully constructed by covalent self-assembly. We assessed their size and morphology by scanning electron microscopy (SEM), TEM, and DLS. (CP)6NS were found to have a strong positive charge, so they could bind to siRNA through electrostatic interactions. Confocal microscopy analysis and cell viability assays showed that (CP)6NS had high cellular internalization efficiency and low cytotoxicity. More importantly, real-time polymerase chain reaction (PCR) and flow cytometry analyses indicated that (CP)6NS-siRNA complexes potently inhibited gene expression and promoted tumor cell apoptosis. These results suggest that (CP)6NS may be a potential siRNA carrier for gene therapy.

Synthesis and Characterization of Palmitoyl-block-poly(methacryloyloxyethyl phosphorylcholine) Polymer Micelles for Anticancer Drug Delivery.

We report the synthesis and characterization of an amphiphilic polymer comprising a hydrophobic palmitoyl (Pal) group and a zwitterionic poly(2-methacryloyloxyethyl phosphorylcholine) (pMPC) block, which is capable of forming micelles as a drug carrier system for delivering hydrophobic anticancer drugs such as doxorubicin (DOX). We hypothesize that the sharp polarity contrast between the Pal domain and the pMPC block would strengthen the micelles and improve the drug loading capacity, while the pMPC shells improve the micelle stability and cellular uptake efficiency. In this study, the Pal-pMPC polymer was characterized and compared with a Pal-poly(ethylene glycol) (Pal-PEG) polymer in terms of their micelle formation, cytotoxicity, and drug loading of DOX. The DOX-loaded Pal-pMPC micelles were further evaluated for the cellular uptake and anticancer activities in cell culture systems including the non-multidrug-resistance HeLa cell line and the multidrug-resistance AT3B-1 cell line. The results showed that the Pal-pMPC polymer had a minimal toxicity. The Pal-pMPC micelles exhibited higher drug loading capacity and enhanced cellular internalization efficiency compared to micelles formed by the Pal-PEG polymer. It was also found that DOX-loaded Pal-pMPC micelles exhibited a more efficient anticancer effect than Pal-PEG micelles in multidrug-resistance cancer cells in an environment with fetal bovine serum.

Geometry of a DNA Nanostructure Influences Its Endocytosis: Cellular Study on 2D, 3D, and in Vivo Systems.

Fabrication of nanoscale DNA devices to generate 3D nano-objects with precise control of shape, size, and presentation of ligands has shown tremendous potential for therapeutic applications. The interactions between the cell membrane and different topologies of 3D DNA nanostructures are crucial for designing efficient tools for interfacing DNA devices with biological systems. The practical applications of these DNA nanocages are still limited in cellular and biological systems owing to the limited understanding of their interaction with the cell membrane and endocytic pathway. The correlation between the geometry of DNA nanostructures and their internalization efficiency remains elusive. We investigated the influence of the shape and size of 3D DNA nanostructures on their cellular internalization efficiency. We found that one particular geometry, i.e., the tetrahedral shape, is more favored over other designed geometries for their cellular uptake in 2D and 3D cell models. This is also replicable for cellular processes like cell invasion assays in a 3D spheroid model, and passing the epithelial barriers in in vivo zebrafish model systems. Our work provides detailed information for the rational design of DNA nanodevices for their upcoming biological and biomedical applications.

Cothreading of Unmodified and Monoazidated β-Cyclodextrins in Polyrotaxanes for Orthogonal Modification of Cell-Penetrating Peptides via Click Chemistry

To establish a method for the orthogonal chemical functionalization of β-cyclodextrin (β-CD)-based polyrotaxane (PRX), unmodified and monoazidated β-CDs (N3-β-CDs) were utilized as cyclic host molecules for synthesizing PRXs. PRXs were successfully synthesized by using N3-β-CD, and the molar ratio of unmodified β-CD and N3-β-CD in the resulting PRXs could be regulated by the feed ratio of unmodified β-CD and N3-β-CD. It was revealed that all PRX molecules contained N3-β-CD, indicating that unmodified and monoazidated β-CDs were cothreaded onto a single polymer axis. Physicochemical characterization of β-CD/N3-β-CD cothreaded PRXs suggested that their crystalline structures were unaffected by the molar ratio of N3-β-CD, whereas their thermal stability was slightly improved with increasing molar ratio of N3-β-CD. To facilitate cellular internalization of PRXs, threaded N3-β-CD in PRXs was modified with cell-penetrating octaarginine (R8) peptides via a copper(I)-catalyzed azide–alkyne click reaction. The cellular internalization efficiency of the R8-modified PRXs increased with the number of R8 peptides modified on PRXs. However, PRXs modified with a large number of R8 peptides precipitated in serum solutions and exhibited high cytotoxicity. PRXs modified with the optimal number of R8 peptides did not exhibit precipitation and showed high cellular uptake compared to unmodified PRXs without cytotoxicity. Consequently, PRXs modified with the optimal number of R8 peptides showed superior cholesterol-reducing effects in fibroblasts derived from patients with Niemann–Pick type C disease. As demonstrated here, the number of modified functional molecules plays a crucial role in the treatment effects of PRXs, and β-CD/N3-β-CD cothreaded PRXs can offer potential for orthogonal postmodification. Thus, β-CD/N3-β-CD cothreaded PRXs could introduce benefits that enable the material applications of PRXs.

ROS-initiated in-situ polymerization of diacetylene-containing lipidated peptide amphiphile in living cells

Recently, in-situ polymerization inside living cells has attracted much attention due to the efficient cellular internalization and elevated drug retention. However, the lack of tracking of the in-situ polymerization process and the unclear effects of polymerization on cellular functions restrict its biomedical applications. Herein, we designed a Y-shaped diacetylene-containing lipidated peptide amphiphile (Y-DLPA1) with positive charges, which underwent in-situ polymerization initiated by reactive oxygen species in the intracellular microenvironment. In comparison, zwitterionic Y-DLPA2 and negatively charged Y-DLPA3 were polymerized in aqueous solution, but cannot polymerize in the intracellular microenvironment. The polymerized Y-DLPA1 with red fluorescence provides a platform to label cells for long-term tracking studies. This polymerization reaction induced tumor cell apoptosis, increased cell viscosity and decreased cell motility, which potentially inhibited tumor metastasis and served as a novel antitumor agent. This work provides a novel strategy to track in-situ polymerization process and modulate cell biofunctions.

Amino-Acid-Encoded Supramolecular Photothermal Nanomedicine for Enhanced Cancer Therapy.

Photothermal nanomedicine based on self-assembly of biological components, with excellent biosafety and customized performance, is vital significance for precision cancer therapy. However, the programmable design of photothermal nanomedicine remains extremely challenging due to the vulnerability and variability of noncovalent interactions governing supramolecular self-assembly. Herein, it is reported that amino acid encoding is a facile and potent means to design and construct supramolecular photothermal nanodrugs with controlled therapeutic activities. It is found that the amount and type of amino acid dominates the assembled nanostructures, structural stability, energy-conversion pathway, and therapeutic mechanism of the resulting nanodrugs. Two optimized nanodrugs are endowed with robust structural integrity against disassembly along with high photothermal conversion efficiency, efficient cellular internalization, and enhanced tumor accumulation, which result in more efficient tumor ablation. This work demonstrates that design based on amino acid encoding offers an unprecedented opportunity for the construction of remarkable photoactive nanomedicines toward cancer diagnostics and therapeutics.

Supramolecular Radiosensitizer Based on Hypoxia-Responsive Macrocycle.

Radiotherapy (RT) has been viewed as one of the most effective and extensively applied curatives in clinical cancer therapy. However, the radioresistance of tumor severely discounts the radiotherapy outcomes. Here, an innovative supramolecular radiotherapy strategy, based on the complexation of a hypoxia‐responsive macrocycle with small‐molecule radiosensitizer, is reported. To exemplify this tactic, a carboxylated azocalix[4]arene (CAC4A) is devised as molecular container to quantitatively package tumor sensitizer banoxantrone dihydrochloride (AQ4N) through reversible host–guest interaction. Benefited from the selective reduction of azo functional groups under hypoxic microenvironment, the supramolecular prodrug CAC4A•AQ4N exhibits high tumor accumulation and efficient cellular internalization, thereby significantly amplifying radiation‐mediated tumor destruction without appreciable systemic toxicity. More importantly, this supramolecular radiotherapy strategy achieves an ultrahigh sensitizer enhancement ratio (SER) value of 2.349, which is the supreme among currently reported noncovalent‐based radiosensitization approach. Further development by applying different radiosensitizing drugs can make this supramolecular strategy become a general platform for boosting therapeutic effect in cancer radiotherapies, tremendously promising for clinical translation.

Enhancing the Endocytosis of Phosphatidylserine-Containing Liposomes through Tim4 by Modulation of Membrane Fluidity.

Phosphatidylserine (PS) is a unique lipid that is recognized by the endogenetic receptor, T-cell immunoglobulin mucin protein 4 (Tim4), and PS-containing liposomes have potential use in therapeutic applications. We prepared PS-containing liposomes of various lipid compositions and examined how lipid membrane fluidity affects PS recognition by Tim4 and the resulting endocytosis efficiency into Hela cells. Surface plasmon resonance and laurdan studies showed that increasing lipid membrane fluidity increased the stability of the PS-Tim4 interaction but hampered the entry of liposomes into cells. These results show that endocytosis efficiency is determined by balancing opposing forces induced by membrane fluidity. We found that inclusion of the zwitterionic helper lipid, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, into liposomes ensured efficient cellular internalization because the presence of this lipid provides an ideal balance of lipid fluidity and Tim4 affinity. The results showed that PS recognition by Tim4 and the resulting endocytosis efficiency can be maximized by modulating the membrane fluidity of liposomes by selecting a zwitterionic helper lipid. This study improves our understanding of how to rationally optimize nanotechnology for targeted drug delivery.

Synergistic Therapy of a Naturally Inspired Glycopolymer-Based Biomimetic Nanomedicine Harnessing Tumor Genomic Instability.

Inspired by natural saccharide-protein complexes, a stimuli-responsive biodegradable and branched glycopolymer-pyropheophorbide-a (Ppa) conjugate (BSP) with saccharide units for cancer therapy is constructed. A linear glycopolymeric conjugate (LSP), a branched glycopolymeric conjugate (BShP) from Ppa with long carbon chains, and a branched conjugate (BHSP) based on poly[N-(2-hydroxypropyl) methacrylamide] (polyHPMA) without saccharide units are prepared as controls. Through structure-activity relationship studies, BSP with a 3D network structure forms stable nanostructures via weak intermolecular interactions, regulating the stacking state of Ppa to improve the singlet oxygen quantum yield and the corresponding photodynamic therapy (PDT) effect. BSP shows high loading of olaparib, and are further coated with tumor cell membranes, resulting in a biomimetic nanomedicine (CM-BSPO). CM-BSPO shows highly efficient tumor targeting and cellular internalization properties. The engulfment of CM-BSPO accompanied with laser irradiation results in a prominent antitumor effect, evidenced by disruption of cell cycles in tumor cells, increased apoptosis and DNA damage, and subsequent inhibition of repair for damaged DNA. The mechanism for the synergistic effect from PDT and olaparib is unveiled at the genetic and protein level through transcriptome analysis. Overall, this biodegradable and branched glycopolymer-drug conjugate could be effectively optimized as a biomimetic nanomedicine for cancer therapy.

A biocompatible superparamagnetic chitosan-based nanoplatform enabling targeted SN-38 delivery for colorectal cancer therapy

7-Ethyl-10-hydroxycamptothecin (SN-38) as a potent anti-tumor candidate, suffers the constraints from its poor water solubility, pH-dependent lactone ring stability and the lack of efficient delivery system without losing its activity. Herein, biocompatible superparamagnetic chitosan-based nanocomplexes complexing with water-soluble polymeric prodrug poly(L-glutamic acid)-SN-38 (PGA-SN-38) was engineered for efficient delivery of SN-38. The manufacturing process of colloidal complexes was green, expeditious and facile, with one-shot addition of PGA-SN-38 into chitosan solution without using any organic solvent or surfactant. Upon introducing ultra-small-size superparamagnetic nanoparticles (~10 nm), the developed magnetic nanocomplexes exhibited significantly boosted tumor-targeted accumulation and efficient cellular internalization under a local magnetic field. Notably, the magnetic nanocomplexes achieved distinctly superior targeting and anti-tumor efficacy in the established xenograft colorectal cancer model of mice, with high tumor suppression rate up to 81%. Therefore, this superparamagnetic chitosan-based nanocomplex system could provide a promising platform for the targeted delivery of SN-38 in colorectal cancer therapy.

Folic acid-conjugated raloxifene-loaded graphene-based nanocarrier: Fabrication, characterization and antitumor screening

Breast cancer is the second leading cause of cancer death among women. The efficacy of most current therapies is compromised by the development of drug resistance and/or lack of target site specificity. Graphene nanoribbons (GNRs) represent one of the emerging graphene-based nanocarriers that show high target specificity and efficient cellular internalization for cancer cells. Herein, we screened the potential antitumor activity of a nanocarrier based on graphene oxide and folic acid and loaded with the selective estrogen receptor modulator, raloxifene hydrochloride (RXF), on breast cancer cells. Oxidized graphene nanoribbons (OGNRs), obtained by longitudinal unzipping technique, were functionalized with folic acid (FA), and were loaded with RXF. The prepared RXF-loaded OGNRs-FA showed a multi-layered structure with loading efficiency and entrapment efficiency of 37% and 56%, respectively. In vitro release revealed a pH-dependent release of RXF from OGNRs. Cytotoxicity screening of RXF-OGNRS-FA was investigated for two breast cancer cell lines; MCF-7 and MDA-MB-231. RXF-loaded OGNRs-FA exerted dose- and time-dependent effects against both breast cancer cells. Confocal microscopy imaging showed that surface functionalization of OGNRs with FA remarkably enhanced cellular uptake of OGNRs by both cell lines relative to non-functional OGNRs. Collectively, OGNRs-FA may represent an appealing nanomaterial that mediates cell-specific drug delivery to breast cancer cells. However, a thorough investigation of the in vivo fate and long-term toxicity of OGNRs-FA remains to be elucidated prior their full utilization in biomedical applications. • Oxidized graphene nanoribbons (OGNRs) represent plausible delivery vehicle for cancer treatment. • Conjugating folic acid to graphene nanoribbons promotes targeted delivery to breast cancer cells. • Raloxifene-loaded graphene nanoribbons exerted potent cytotoxicity against breast cancer cells. • Folic acid-functionalized OGNRs significantly enhanced cellular uptake of OGNRs by cancer cells.

Engineering of an EpCAM-targeting cyclic peptide to improve the EpCAM-mediated cellular internalization and tumor accumulation of a peptide-fused antibody

Fusion of a target-specific peptide to a full-length antibody (Ab) can result in a peptide–Ab fusion protein with additional specificity and enhanced activity. We recently developed an intracellular pan-RAS–targeting cytosol-penetrating antibody, RT22-ep59, in which a tumor-specific targeting ability was achieved via the fusion of an epithelial cell adhesion molecule (EpCAM) targeting cyclic peptide (ep133). Here, the aim was to enhance EpCAM-mediated endocytosis and tumor accumulation of the peptide-fused RAS-targeting Ab. Accordingly, we engineered a cyclic peptide (from ep133) that has stronger affinity for EpCAM by using yeast surface display technology and then rationally designed cyclic peptides in the Ab-fused form to enhance colloidal stability. The finally engineered EpCAM-targeting cyclic peptide (ep6)–fused Ab, ep6Ras37, has ∼10-fold stronger affinity (KD ≈ 1.9 nM) for EpCAM than that of RT22-ep59, without deterioration of biophysical properties. Compared with the parental antibody (RT22-ep59), ep6Ras37 more efficiently reached the cytosol of EpCAM-expressing cells and showed greater preferential tumor homing and accumulation in mice bearing EpCAM-expressing LoVo xenograft tumors. Thus, the high-affinity EpCAM-targeting peptide ensures efficient cellular internalization and better tumor accumulation of the peptide-fused Ab.

Polymeric micelles functionalized with cell penetrating peptides as potential pH-sensitive platforms in drug delivery for cancer therapy: A review

Improving the intrinsic pharmacodynamics or pharmacokinetic of drugs is regarded as a crucial step in developing potential drug therapies. In this context, the development of nanostructured-based drug delivery systems (DDSs) responsive to exogenous or endogenous stimuli demonstrate an outstanding platform to clinical applications. Polymeric micelles (PM)-based DDSs are often modified with selective short peptides like cell penetrating peptides (CPPs) for highly efficient cellular internalization. In this review, we discussed recent achievements in the design and development of nanoscale pH-responsive micelles modified with CPPs to provide controlled drug release in the tumor cells. An overview on the application of micelles, especially PMs as promising engineered nanocarriers to combine pharmaceutical agents with CPPs are presented. Afterwards, the peptide-based transfection method as one of the promising approaches for drug delivery and therapeutic agents was explained. Subsequently, conjugation of pH-responsive micelles with CPPs as a novel DDS in cancer therapy was surveyed. Finally, the current challenges and future application of stimuli-sensitive nanoplatforms conjugated with CPP were discussed. In conclusion, the application of controlled delivery using pH-sensitive PMs-based DDSs conjugated with peptides can be considered as a potential approach to improve the therapeutic index of anticancer drugs.