Thiol-maleimide Click Reaction Research Articles

Nanoplatelets modified with RVG for targeted delivery of miR-375 and temozolomide to enhance gliomas therapy

Gliomas are one of the most frequent primary brain tumors and pose a serious threat to people’s lives and health. Platelets, a crucial component of blood, have been applied as drug delivery carriers for disease diagnosis and treatment. In this study, we designed engineered nanoplatelets for targeted delivery of therapeutic miR-375 and temozolomide (TMZ, a first-line glioma treatment agent) to enhance glioma therapy. Nanoplatelets were prepared through mild ultrasound, TMZ and miR-375 were co-loaded through ultrasound and electrostatic interactions, respectively, to combine chemotherapy with gene therapy against glioma. To improve the blood brain barrier (BBB) crossing efficiency and glioma targeting ability, the nanoplatelets were modified with central nervous system-specific rabies viral glycoprotein peptide (RVG) through thiol-maleimide click reaction. The RVG modified nanoplatelets co-loaded TMZ and miR-375 (NR/TMZ/miR-375) not only inherited the good stability and remarkable biocompatibility of platelets, but also promoted the cellular uptake and penetration of glioma tissues, and effectively induced cell apoptosis to enhance the therapeutic effect of drugs. In vivo studies showed that NR/TMZ/miR-375 significantly increased the circulation time of TMZ, and exhibited superior combined antitumor effects. In summary, this multifunctional ‘natural’ nanodrug delivery system provides a potent, scalable, and safety approach for platelet-based combined cancer chemotherapy and gene therapy.

Development of 5-Fluorouracil/pH-Responsive Adjuvant-Embedded Extracellular Vesicles for Targeting αvβ3 Integrin Receptors in Tumors.

To selectively target and treat murine melanoma B16BL6 tumors expressing αvβ3 integrin receptors, we engineered tumor-specific functional extracellular vesicles (EVs) tailored for the targeted delivery of antitumor drugs. This objective was achieved through the incorporation of a pH-responsive adjuvant, cyclic arginine-glycine-aspartic acid peptide (cRGD, serving as a tumor-targeting ligand), and 5-fluorouracil (5-FU, employed as a model antitumor drug). The pH-responsive adjuvant, essential for modulating drug release, was synthesized by chemically conjugating 3-(diethylamino)propylamine (DEAP) to deoxycholic acid (DOCA, a lipophilic substance capable of integrating into EVs' membranes), denoted as DEAP-DOCA. The DOCA, preactivated using N-(2-aminoethyl)maleimide (AEM), was chemically coupled with the thiol group of the cRGD-DOCA through the thiol-maleimide click reaction, resulting in the formation of cRGD-DOCA. Subsequently, DEAP-DOCA, cRGD-DOCA, and 5-FU were efficiently incorporated into EVs using a sonication method. The resulting tumor-targeting EVs, expressing cRGD ligands, demonstrated enhanced in vitro/in vivo cellular uptake specifically for B16BL6 tumors expressing αvβ3 integrin receptors. The ionization characteristics of the DEAP in DEAP-DOCA induced destabilization of the EVs membrane at pH 6.5 through protonation of the DEAP substance, thereby expediting 5-FU release. Consequently, an improvement in the in vivo antitumor efficacy was observed for B16BL6 tumors. Based on these comprehensive in vitro/in vivo findings, we anticipate that this EV system holds substantial promise as an exceptionally effective platform for antitumor therapeutic delivery.

Mucoadhesive Thiolated Hyaluronic Acid/Pluronic F127 Nanogel Formation via Thiol-Maleimide Click Reaction for Intravesical Drug Delivery.

The development of nanocarriers to prolong the residence time and enhance the permeability of chemotherapeutic drugs on bladder mucosa is important in the postsurgery treatment of superficial bladder cancers (BCs). Here, the mucoadhesive HA-SH/PF127 nanogels composed of a temperature-sensitive Pluronic F127 (PF127) core and thiolated hyaluronic acid (HA-SH) shell were prepared by the emulsification/solvent evaporation method. The nanogels were constructed through the thiol-maleimide click reaction in the HA-SH aqueous side of the oil-water interface and self-oxidized cross-linking thiols between HA-SH. The HA-SH/PF127 nanogels prepared at different thiol-to-maleimide group molar ratios, water-to-oil volume ratios, and cross-linking reaction times were characterized regarding hydrodynamic diameter (Dh) and zeta potential (ζ), and the optimal formulation was obtained. The excellent mucoadhesive properties of the HA-SH/PF127 nanogels were evaluated by using the mucin particle method. Doxorubicin (DOX) was encapsulated in the PF127 core of DOX@HA-SH/PF127 nanogels with a high loading efficiency (87.5%) and sustained release from the nanogels in artificial urine. Ex vivo studies on porcine bladder mucosa showed that the DOX@HA-SH/PF127 nanogels enhanced the penetration of the DOX into the bladder mucosa without disrupting the mucus structure or the bladder tissue. A significant dose-dependent cytotoxic effect of DOX@HA-SH/PF127 nanogels on both T24 and MB49 cells was observed. The present study demonstrates that the mucoadhesive HA-SH/PF127 nanogels are a promising intravesical drug delivery system for superficial BC therapy.

Star-Shaped Peptide-Polymer Hybrids as Fast pH-Responsive Supramolecular Hydrogels.

Significant challenges have gone into the design of smart hydrogels, with numerous potential applications in the industrial, cosmetic, and biomedical fields. Herein, we report the synthesis of novel 4-arm self-assembling peptide-polyethylene glycol (PEG) hybrid star-shaped polymers and their comprehensive hydrogel properties. β-sheet-forming oligopeptides with alternating hydrophobic Leu/ionizable Glu repeats and Cys residues were successfully conjugated to 4-arm PEG via a thiol-maleimide click reaction. The hybrid star-shaped polymers demonstrated good cytocompatibility and reversible β-sheet (lightly acidic pH)-to-random coil (neutral and basic pH) transition in dilute aqueous solutions. At increasing polymer concentrations up to 0.5 wt %, the star-shaped polymers formed transparent hydrogels with shear-thinning and self-healing behaviors via β-sheet self-assembly, as well as a conformation-dependent gel-sol transition. Interestingly, the star-shaped polymers responded rapidly to pH changes, causing gelation to occur rapidly within a few seconds from the change in pH. Hydrogel characteristics could be modulated by manipulating the length and net charge of the peptide blocks. Furthermore, these star-shaped polymers served as satisfactory network scaffolds that could respond to dynamic environmental changes in the pH-oscillation system, owing to their excellent gelation capability and pH sensitivity. As such, they are highly favorable for diverse applications, such as pH-responsive controlled release.

Selective detection of Aeromonas spp. by a fluorescent probe based on the siderophore amonabactin

Amonabactins, the siderophores produced by some pathogenic bacteria belonging to Aeromonas genus, can be used for the preparation of conjugates to be imported into the cell using their specific transport machinery. Herein, we report the design and synthesis of a new amonabactin-based fluorescent probe by conjugation of the appropriate amonabactin analogue to sulforhodamine B (AMB-SRB) using a thiol-maleimide click reaction. Growth promotion assays and fluorescence microscopy studies demonstrated that the AMB-SRB fluorescent probe was able to label the fish pathogenic bacterium A. salmonicida subsp. salmonicida through its outer membrane transport (OMT) protein FstC. The labelling of other Aeromonas species, such as the human pathogen A. hydrophila, indicates that this probe can be a very useful molecular tool for studying the amonabactin-dependent iron uptake mechanism. Furthermore, the selective labelling of A. salmonicida and other Aeromonas species in presence of other fish pathogenic bacteria, suggest the potential application of this probe for detection of Aeromonas in water and other fish farming samples through fluorescence assays.

Preparation and Insights of Smart Foams with Phototunable Foamability Based on Azobenzene-Containing Surfactants.

Smart foams with tunable foamability exhibit superb applications in many fields such as colloidal and interface science. Herein, we have synthesized an azobenzene-containing surfactant with excellent photoresponsiveness by a simple thiol-maleimide click reaction between thioglycolic acid and 4-(N-maleimide) azobenzene (MAB). The structure and the photoresponsive behavior of the novel surfactant are characterized. Depending on the solution concentration, the synthesized surfactant demonstrated various speeds for the trans/cis photoisomerization varying from 9 to 24 s for the given concentration range and excellent reversible photoisomerization cycling stability (more than 20 cycles) upon light irradiation. Based on these conformational switches, a series of phototriggered obvious surface properties (e.g., critical micelle concentration (CMC), surface tension (γ), and surface excess concentration (Γ)) changes of the surfactant are achieved. More specifically, the smart foam system with tunable foamability is realized. As-formed smart foams with rapid photocontrolled reversible foaming/defoaming transition and excellent cycling stability make them very attractive candidates for applications in wastewater treatment, green textile, oil extraction, and emulsification.

64Cu-Labeled Ubiquitin for PET Imaging of CXCR4 Expression in Mouse Breast Tumor.

Ubiquitin has been recently identified as a chemokine receptor 4 (CXCR4) natural ligand, offering great potential for positron emission computed tomography (PET) imaging of CXCR4 expression. This study reports the preparation and evaluation of (64Cu)-radiolabeled ubiquitin for CXCR4 imaging. The ubiquitin was first fused with a C-terminal GGCGG sequence, and the resulting recombinant ubiquitin derivative UbCG4 was then functionalized with the trans-cyclooctene (TCO) moiety via thiol–maleimide click reaction, followed by 64Cu-radiolabeling through the TCO/Tz (tetrazine)-based Diels–Alder click reaction. In the prepared in vitro studies, the prepared (64Cu)-UbCG4 showed significantly higher specific uptakes in the 4T1 breast cancer cells compared with the uptakes in the CXCR4-knockdown 4T1 cells. In the in vivo evaluation in the 4T1-xenograft mouse model, (64Cu)-UbCG4 demonstrated a similar tumor uptake but much lower backgrounds compared with 64Cu-labeled AMD3465. These results suggested that (64Cu)-UbCG4 could serve as a potent PET tracer for the noninvasive imaging of CXCR4 expression in tumors.

Cell-penetrating peptide modified AIE polymeric nanoparticles by miniemulsion polymerization and application for cell fluorescence imaging

AIE polymeric nanoparticles surface-modified with HIV Tat peptides for cell fluorescence imaging are efficiently prepared through miniemulsion polymerization, carbodiimide reaction, and thiol-maleimide click reaction in sequence.

PH-modulated ion-current rectification in a cysteine-functionalized glass nanopipette

The unique properties of nanopores depend largely on the functionalization of the pore surface, which has always been a great challenge. In this study, a facile method for functionalizing the surface of a glass nanopipette to establish thiol-based linkage without involving Au film coating was successfully developed by utilizing the thiol–maleimide click reaction which works rapidly and with high specificity. Cysteine was employed as a model molecule to functionalize the nanopipette, resulting in pH-modulated ion-current rectification behavior which showed a good reproducible response to pH. Due to the widespread existence of thiol groups in nature, this nanopipette functionalization method has the potential and versatility to be used in a wide range of applications, including mimicking biological ion channels and fabricating sensing devices.

Antibacterial finishing of cotton fabrics based on thiol-maleimide click chemistry

Cotton fabrics with durable antibacterial properties were prepared by a novel method of thiol-maleimide click reaction. Firstly, 3-Mercaptopropyltriethoxysilane was utilized as a modifier to generate thiol groups on the cotton surface. Then, these thiol groups reacted with N-phenyl-male-imide through thiol-maleimide click chemistry. The surface morphology of the treated fabrics and the reaction mechanism were characterized by FT-IR, Raman, EDS, XPS and SEM. Antibacterial activities, mechanical properties and thermal performance of treated cotton fabrics were also investigated. The Escherichia coli antibacterial rate of treated cotton was 99.56% and the Staphylococcus aureus antibacterial rate of treated cotton was 98.91%, with only a slight decrease after 10 cycles of standard washing, to 88.69 and 87.66%, respectively. These results demonstrated that this treatment effectively endowed cotton fabrics with durable antibacterial properties due to the chemical bonding formed between the antibacterial agent and the substrate. In addition, the treated cotton fabrics maintained good mechanical properties and thermal stability.

Supramolecular Interfacial Polymerization: A Controllable Method of Fabricating Supramolecular Polymeric Materials

AbstractA new method of supramolecular polymerization at the water–oil interface is developed. As a demonstration, an oil‐soluble supramonomer containing two thiol end groups linked by two ureidopyrimidinone units and a water‐soluble monomer bearing two maleimide end groups are employed. Supramolecular interfacial polymerization can be implemented by a thiol–maleimide click reaction at the water–chloroform interface to obtain supramolecular polymeric films. The glass transition temperature of such supramolecular polymers can be well‐tuned by simply changing the polymerization time and temperature. It is highly anticipated that this work will provide a facile and general approach to realize control over supramolecular polymerization by transferring the preparation of supramolecular polymers from solutions to water–oil interfaces and construct supramolecular materials with well‐defined properties.

Supramolecular Interfacial Polymerization: A Controllable Method of Fabricating Supramolecular Polymeric Materials.

A new method of supramolecular polymerization at the water–oil interface is developed. As a demonstration, an oil‐soluble supramonomer containing two thiol end groups linked by two ureidopyrimidinone units and a water‐soluble monomer bearing two maleimide end groups are employed. Supramolecular interfacial polymerization can be implemented by a thiol–maleimide click reaction at the water–chloroform interface to obtain supramolecular polymeric films. The glass transition temperature of such supramolecular polymers can be well‐tuned by simply changing the polymerization time and temperature. It is highly anticipated that this work will provide a facile and general approach to realize control over supramolecular polymerization by transferring the preparation of supramolecular polymers from solutions to water–oil interfaces and construct supramolecular materials with well‐defined properties.

Optimization of Curcumin-Loaded PEG-PLGA Nanoparticles by GSH Functionalization: Investigation of the Internalization Pathway in Neuronal Cells.

One major challenge in the field of nanotherapeutics is to increase the selective delivery of cargo to targeted cells. Using polylactic-co-glycolic acid (PLGA), we recently highlighted the importance of polymer composition in the biological fate of the nanodrug delivery systems. However, the route of internalization of polymeric nanoparticles (NPs) is another key component to consider in the elaboration of modern and targeted devices. For that purpose, herein, we effectively synthesized and characterized glutathione-functionalized PLGA-nanoparticles (GSH-NPs) loaded with curcumin (GSH-NPs-Cur), using thiol-maleimide click reaction and determined their physicochemical properties. We found that GSH functionalization did not affect the drug loading efficiency (DLE), the size, the polydispersity index (PDI), the zeta potential, the release profile, and the stability of the formulation. While being nontoxic, the presence of GSH on the surface of the formulations exhibits a better neuroprotective property against acrolein. The neuronal internalization of GSH-NPs-Cur was higher than free curcumin. In order to track the intracellular localization of the formulations, we used a covalently attached rhodamine (PLGA-Rhod), into our GSH-functionalized matrix. We found that GSH-functionalized matrix could easily be taken up by neuronal cells. Furthermore, we found that GSH conjugation modifies the route of internalization enabling them to escape the uptake through macropinocytosis and therefore avoiding the lysosomal degradation. Taken together, GSH functionalization increases the uptake of formulations and modifies the route of internalization toward a safer pathway. This study shows that the choice of ideal ligand to develop NPs-targeting devices is a crucial step when designing innovative strategy for neuronal cells delivery.

Determination of cysteine using near-infrared diffuse reflectance spectroscopy with enrichment via thiol-maleimide click reaction

Enrichment technique has been proved to be an efficient way to make the near-infrared diffuse reflectance spectroscopy(NIRDRS) suitable for micro analysis. However, low selectivity presented by conventional enrichment methods makes the quantitative analysis easy to be affected by the coexisting components. In this study, a specific enrichment method with chemical bonding via thiol-maleimide click reaction was used to achieve the reduction of the interferences. Taking cysteine as the analyzing target, maleimide-functionalized SiO2 nanoparticles were prepared for the enrichment of cysteine. Then determination of cysteine in aqueous solution and human serum was studied using the partial least squares model built from the NIRDRS spectra of the adsorbate. The results show that the concentration that can be quantitatively detected is as low as 2.0 μg/mL, and the correlation coefficient(R) between the reference and predicted concentration is 0.9871 for the validation samples. The recoveries are in the range of 89.5%―113.8% for human serum samples in the concentration range of 0―16.2 μg/mL.

Functionalizing Liposomes with anti-CD44 Aptamer for Selective Targeting of Cancer Cells.

CD44 receptor protein is found to be overexpressed by many tumors and is identified as one of the most common cancer stem cell surface markers including tumors affecting colon, breast, pancreas, and head and neck, making this an attractive receptor for therapeutic targeting. In this study, 2'-F-pyrimidine-containing RNA aptamer (Apt1), previously selected against CD44, was successfully conjugated to the surface of PEGylated liposomes using the thiol-maleimide click reaction. The conjugation of Apt1 to the surface of liposomes was confirmed by the change in size and zeta potential and by migration on agarose gel electrophoresis. The binding affinity of Apt1 was improved after conjugation compared to free-Apt1. The cellular uptake for Apt1-Lip was tested by flow cytometry and confocal imaging using the two CD44(+) cell lines, human lung cancer cells (A549) and human breast cancer cells (MDA-MB-231), and the CD44(-) cell line, mouse embryonic fibroblast cells (NIH/3T3). The results showed higher sensitivity and selectivity for Apt1-Lip compared to the blank liposomes (Mal-Lip). In conclusion, we demonstrate a successful conjugation of anti-CD44 aptamer to the surface of liposome and binding preference of Apt1-Lip to CD44-expressing cancer cells and conclude to a promising potency of Apt1-Lip as a specific drug delivery system.

Vapor-based synthesis of maleimide-functionalized coating for biointerface engineering

Chemical vapor deposition polymerization was used to deposit a novel maleimide-functionalized poly-p-xylylene coating on various substrates. The coated substrates are readily able to undergo thiol-maleimide click reaction under mild conditions. Applications using this coating technology are highlighted in low-protein-fouling modification as well as manipulated attachments and growth of bovine arterial endothelial cells.