Cu(I)-catalyzed Azide-alkyne Cycloaddition Research Articles

Near-Infrared Light Dual-Promoted Heterogeneous Copper Nanocatalyst for Highly Efficient Bioorthogonal Chemistry in Vivo

Owing to better stability and biosafety, heterogeneous Cu nanoparticles (CuNPs) have been put forward as a promising candidate to complete the Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. However, the inherent poor activity of Cu(0) deterred its wide bioapplication. Herein, we employed near-infrared (NIR) light to dual-promote the CuAAC reaction of a biocompatible heterogeneous copper nanocatalyst through photodynamic and photothermal effects in vitro and in vivo. Specifically, the photodynamic activity could promote the conversion of Cu(0) to Cu(I) to accelerate the catalytic process of CuAAC. The high photothermal conversion efficiency (η = 50.6%) could increase the local temperature, further promoting the whole reaction. Then, a drastically increased reaction rate in a living system ranging from cells to nematodes was achieved in our system. Meanwhile, the better antitumor efficacy has determined with in vivo tumor therapy experiments.

Application of PEG400 in the one-pot synthesis of 7-[4-alkyl- or (hetero)aryl-1H-1,2,3-triazol-1-yl]thieno[3,2-b]pyridines via SNAr and Cu(I)-Catalyzed Azide-Alkyne Cycloaddition and preliminary evaluation of their anti-tumour activity

Several novel 7-[4-alkyl- or (hetero)aryl-1H-1,2,3-triazol-1-yl]thieno[3,2-b]pyridines were prepared in good to high yields, using the environmentally friendly solvent PEG400 in a one-pot procedure from 7-chlorothieno[3,2-b]pyridine to form the corresponding azide via SNAr with NaN3, followed by Cu(I)-catalyzed Azide-Alkyne Cycloaddition (CuAAC) using different types of alkynes. This one-pot reaction in PEG400 starting from a halogenated heteroaromatic system is reported for the first time and demonstrated a wide scope of application for alkynes. Preliminary anti-tumour activity on human tumour cell lines using the prepared 1,4-di(hetero)aryl-1,2,3-triazoles was evaluated, together with their toxicity in non-tumour cells. Among the tested compounds the most promising one was a 2-ethynylpyridine derivative.

Targeting of Immune Cells with Trimannosylated Liposomes

AbstractDendritic cells (DCs) are a compelling target in cancer immunotherapy as they initialize strong antigen‐specific immune responses. Drug delivery systems (DDSs) such as liposomes provide the opportunity to deliver antigens and immunostimulatory molecules to DCs, which in turn initiate an antigen‐specific immune response. To address predominantly DCs, DDSs need to be equipped with targeting moieties. This study evaluates liposomes, bearing the oligosaccharide trimannose on their surface, for their ability to address DCs in vitro and in vivo. Trimannose as a saccharidic structure is known to be recognized by receptors on the surface of DCs. To obtain trimannosylated liposomes, azide‐bearing trimannose is coupled to alkyne‐functionalized hyperbranched polyglycerol (hbPG) with a bis(hexadecyl)glycerol (BisHD) anchor in a Cu(I)‐catalyzed alkyne‐azide cycloaddition (CuAAC). To enable tracking of the liposomes in vivo, the trimannosylated BisHD‐hbPG lipids are radiolabeled with 18F in a CuAAC. Subsequently, liposomes are produced via the thin‐film hydration method followed by extrusion. The behavior of the trimannosylated liposomes is evaluated in in vitro cell binding assays and in vivo µPET and ex vivo biodistribution studies in healthy C57BL/6 mice and the results are compared to similar liposomes not bearing trimannose on their surface.

Metal-Free Synthesis of Functional 1-Substituted-1,2,3-Triazoles from Ethenesulfonyl Fluoride and Organic Azides.

The boom in growth of 1,4-disubstituted triazole products, in particular, since the early 2000's, can be largely attributed to the birth of click chemistry and the discovery of the CuI -catalyzed azide-alkyne cycloaddition (CuAAC). Yet the synthesis of relatively simple, albeit important, 1-substituted-1,2,3-triazoles has been surprisingly more challenging. Reported here is a straightforward and scalable click-inspired protocol for the synthesis of 1-substituted-1,2,3-triazoles from organic azides and the bench stable acetylene surrogate ethenesulfonyl fluoride (ESF). The new transformation tolerates a wide selection of substrates and proceeds smoothly under metal-free conditions to give the products in excellent yield. Under controlled acidic conditions, the 1-substituted-1,2,3-triazole products undergo a Michael addition reaction with a second equivalent of ESF to give the unprecedented 1-substituted triazolium sulfonyl fluoride salts.

Metal‐Free Synthesis of Functional 1‐Substituted‐1,2,3‐Triazoles from Ethenesulfonyl Fluoride and Organic Azides

AbstractThe boom in growth of 1,4‐disubstituted triazole products, in particular, since the early 2000’s, can be largely attributed to the birth of click chemistry and the discovery of the CuI‐catalyzed azide–alkyne cycloaddition (CuAAC). Yet the synthesis of relatively simple, albeit important, 1‐substituted‐1,2,3‐triazoles has been surprisingly more challenging. Reported here is a straightforward and scalable click‐inspired protocol for the synthesis of 1‐substituted‐1,2,3‐triazoles from organic azides and the bench stable acetylene surrogate ethenesulfonyl fluoride (ESF). The new transformation tolerates a wide selection of substrates and proceeds smoothly under metal‐free conditions to give the products in excellent yield. Under controlled acidic conditions, the 1‐substituted‐1,2,3‐triazole products undergo a Michael addition reaction with a second equivalent of ESF to give the unprecedented 1‐substituted triazolium sulfonyl fluoride salts.

An ultrasensitive fluorescent aptasensor based on truncated aptamer and AGET ATRP for the detection of bisphenol A.

Given the gigantic harmfulness of bisphenol A (BPA), a novel and ultrasensitive aptasensor, which employs the truncated BPA aptamer, click chemistry, and activators generated by electron transfer for atom transfer radical polymerization (AGET ATRP), was developed herein for the quantitative determination of BPA. Firstly, hairpin DNAs (hairpins) with a thiol at the 5' end and an azide group at the 3' end were conjugated with aminated magnetic beads (MBs) through heterobifunctional cross-linkers. BPA truncated aptamer (ssDNA-A) hybridizes with its complementary single-stranded DNA (ssDNA-B) to form double-stranded DNA. In the presence of BPA, ssDNA-A specifically captures BPA, and then ssDNA-B is released. Subsequently, the ssDNA-B hybridizes with hairpins to expose the azide group near the surface of the MBs. Then, propargyl-2-bromoisobutyrate (PBIB), the initiator of AGET ATRP containing alkynyl group, was conjugated with azide group of hairpins via the Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC). Consequently, a large number of fluorescein-o-acrylate (FA) were introduced to the MBs through AGET ATRP, resulting in that the fluorescence intensity was increased dramatically. Obviously, the fluorescence intensity was especially sensitive to the change of BPA concentration, and this method can be used in quantitative determination of BPA. Under optimal conditions, a broad liner range from 100fM to 100nM and a low limit of detection (LOD) of 6.6fM were obtained. Moreover, the method exhibits not only excellent specificity for BPA detection over BPA analogues but high anti-interference ability in real water sample detection, indicating that it has huge application prospect in food safety and environment monitoring.

Micelles with Cyclic Poly(ε-caprolactone) Moieties: Greater Stability, Larger Drug Loading Capacity, and Slower Degradation Property for Controlled Drug Release.

Polymer topology exerts a significant effect on its properties and performance for potential applications. Cyclic topology and its derived structures have been recently shown to outperform conventional linear analogues for drug delivery applications. However, an amphiphilic tadpole-shaped copolymer consisting of a cylic hydrophobic moiety has rarely been explored. For this purpose, a tadpole-shaped amphiphilic diblock copolymer of poly(ethylene oxide)-b-(cyclic poly(ε-caprolactone)) (mPEG-b-cPCL) was synthesized successfully via ring-opening polymerization (ROP) of ε-CL using a mPEG-based macroinitiator with both a hydroxyl and an azide termini and subsequent intrachain Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAc) click cyclization. A comparison study on the self-assembly behaviors, in vitro drug loading and drug release profiles, and degradation properties of the resulting mPEG-b-cPCL (C) with those of the linear counterpart (mPEG-b-PCL, L) revealed that mPEG-b-cPCL micelles are a better formulation than the micelles formed by the linear counterparts in terms of micelle stability, drug loading capacity, and the degradation property. Interestingly, compared to the single degradation of L, C exhibited a slower two-stage degradation process including the topological change from tadpole shape to linear conformation and the subsequent degradation of a linear polymer. This study therefore uncovered the topological effect of a hydrophobic moiety on the properties of the self-assembled micelles and developed a complementary alternative to enhance the micelle stability by introducing a cyclic hydrophobic segment.

Ultrasensitive fluorescent detection of HTLV-II DNA based on magnetic nanoparticles and atom transfer radical polymerization signal amplification

Human T-lymphotropic virus type II (HTLV-II) is a crucial retrovirus that is closely associated with a variety of human diseases. Herein, an ultrasensitive fluorescent HTLV-II DNA detection strategy was developed for the first time based on magnetic nanoparticles (MNPs) and atom transfer radical polymerization (ATRP) amplification. In this approach, hairpin DNA probes (pDNA) labelled with 5′ thiol and 3′ azide group terminally were immobilized on amino group modified MNPs surface through sulfo-N-succinimidyl-4-maleimidobutyrate sodium salt (sulfo-GMBS) cross-linkers. In the presence of target DNAs (tDNA), pDNA hybridized with tDNA to form double-stranded DNA, and therefore its azide group was away from the MNPs surface. Subsequently, to initiate ATRP reaction, initiators were introduced into the pDNA by a Cu (I)-catalyzed alkyne-azide cycloaddition (CuAAC). Then, large numbers of 9-anthracenylmethyl methacrylate polymer (pAMMA) were successfully labelled on the MNPs surface, resulting in significant amplification of the fluorescence signal. Under optimized conditions, the fluorescence signal was proportional to the logarithm of the concentration of tDNA over the range from 1 fM to 1 nM, with a detection limit of 0.22 fM. Moreover, this strategy was capable of discriminating mismatched bases and detecting HTLV-II DNA in human serum samples. By virtue of the high sensitivity, selectivity, simplicity and economy, this ultrasensitive biosensor demonstrates great potential for biomedical research and early clinical diagnosis.

Recyclable Cu nanoparticle catalyzed azide-alkyne click polymerization

The Cu(I)-catalyzed alkyne-azide cycloaddition (CuAAC) has been developed into a powerful polymerization reaction for the synthesis of new polytriazoles with versatile properties. However, research on recyclable and reusable copper catalyst for click polymerization to meet the requirement of green chemistry was rarely reported. Copper nanoparticles were reported to be capable catalysts for CuAAC. Replacing conventional copper catalyst with copper nanoparticles may realize the recycle and reuse of the copper catalyst in click polymerization. In this paper, copper nanoparticles were prepared and used as an effective catalyst for click polymerization, and soluble polytriazoles with high molecular weights were obtained in excellent yields under optimized reaction conditions. Importantly, the copper nanoparticles can be recycled and reused for up to 11 times for the click polymerization. Moreover, introducing aggregation-induced emission (AIE)-active moiety of tetraphenylethylene into the monomers makes the resultant polymers retain the AIE feature. This work not only provides an efficient recyclable catalytic system for the azide-alkyne click polymerization, but also might inspire polymer chemists to use recyclable copper species to catalyze other polymerizations.

On resin click-chemistry-mediated synthesis of novel enkephalin analogues with potent anti-nociceptive activity

Here, we report the chemical synthesis of two DPDPE analogues 7a (NOVA1) and 7b (NOVA2). This entailed the solid-phase synthesis of two enkephalin precursor chains followed by a CuI-catalyzed azide-alkyne cycloaddition, with the aim of improving in vivo analgesic efficacy versus DPDPE. NOVA2 showed good affinity and selectivity for the μ-opioid receptor (KI of 59.2 nM, EC50 of 12.9 nM, EMax of 87.3%), and long lasting anti-nociceptive effects in mice when compared to DPDPE.

“A~A+B3~” strategy to construct redox-responsive core-crosslinked copolymers as potential drug carrier

Redox-responsive core-crosslinked copolymers were synthesized based on poly(ε-caprolactone) (PCL) and poly(ethylene glycol) (PEG) via “A~A + B3~” strategy and Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. It is a feasible method for large-scale preparing redox-responsive core-crosslinked copolymers by crosslinking PCL-based disulfide bond-containing A-PCL-A macromonomers and PEG-based telechelic B3~ macromonomers. The core-crosslinked copolymers can form stable micelles through self-assembly in aqueous solution and exhibit reduction-cleavage responsibility in reductive medium. Further, their doxorubicin (DOX)-loaded micelles exhibit desirable reduction-triggered release performance in vitro. The cellular proliferation inhibition against A549 cells indicates their enhanced anticancer activity in comparison to free DOX. The redox-responsive core-crosslinked copolymers are expected to be used in smart delivery vehicles of anti-cancer drugs.

Hairpin probes based click polymerization for label-free electrochemical detection of human T-lymphotropic virus types II

A novel, simple, and label-free amplification electrochemical impedance method for quantitative detection of Human T-lymphotropic virus types II(HTLV-II) via click chemistry-mediated of hairpin DNA probes (hairpins) with polymers was developed. The hairpins were firstly attached to the gold electrode surface by an S-Au bond, the azido terminals of hairpins were close to the electrode surface, which make it difficult to be approached. After hybridizing with HTLV-II, the hairpins were unfolded and experienced a big configuration change, which made the azido terminals of the hairpins available to conjugate with alkynyl-containing polymer, called P(DEB-DSDA), formed by 1,4-diacetylenebenzene (DEB) and 4,4′-Diazido-2,2′-stilbenedisulfonic acid disodium salt tetrahydrate (DSDA) via Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC). With amount of P(DEB-DSDA) conjugated with the hairpin probes via click polymerization, its electrochemical signal can have a great amplification. Under optimized experimental conditions, this new probe showed a low detection limit of 0.171 pM with a good liner in the range of 1 pM-1 nM. Meanwhile, the biosensor also exhibited good selectivity and reliability in detection of real serum samples, indicating that it has great application potential in clinical DNA diagnosis and detection.

Crystallization in PEG networks: The importance of network topology and chain tilt in crystals

We synthesized poly(ethylene glycol) (PEG) networks by Cu(I) catalyzed azide-alkyne cycloaddition (CuAAC) using three-arm star-PEG with azide end groups and linear PEG oligomers of different molar masses with alkyne end groups. After extensive characterization of the networks by 13C MAS and 1H multiple-quantum (MQ) NMR spectroscopy, their crystallization behavior is studied by differential scanning calorimetry (DSC), temperature dependent small and wide angle X-ray scattering (SAXS, WAXS) as well as 1H FID NMR. Since CuAAC introduces 1,4-disubstituted 1,2,3-triazole (TR) rings as chain defects into the network, we compare our results with those obtained from linear PEG which contained a purposely introduced TR ring in the middle of the polymer chain. PEG network chains are able to form the characteristic 72 helices upon cooling to −20 °C and crystallize into the monoclinic unit cell in complete analogy to the homopolymer. The crystallinity of the networks, though it is lower as compared to the linear samples, reached values of about 30–50%. Given the fact that the network chains are highly constrained in their configurational freedom, this is very surprising. However, cross-links and the TR defects have only a minor effect on the molecular dynamics of the PEG chains in the crystals since the characteristic αc-relaxation process (helix jumps) is also detected in the PEG networks by analysis of the 1H FIDs. The SAXS data reveal that randomly arranged stacks of lamellae are formed upon crystallization and the chains are tilted in the crystals at large angles of about 40°–55°. Supported by all experimental data, we propose a model to explain the formation of polymer crystals in the networks and concluded that primary and secondary loops which extent the network chains beyond the length of the linear precursors play an essential role for the crystallization.

Stable Molecular Surface Modification of Nanostructured, Mesoporous Metal Oxide Photoanodes by Silane and Click Chemistry.

Binding functional molecules to nanostructured mesoporous metal oxide surfaces provides a way to derivatize metal oxide semiconductors for applications in dye-sensitized photoelectrosynthesis cells (DSPECs). The commonly used anchoring groups, phosphonates and carboxylates, are unstable as surface links to oxide surfaces at neutral and high pH, leading to rapid desorption of appended molecules. A synthetically versatile molecular attachment strategy based on initial surface modification with a silyl azide followed by click chemistry is described here. It has been used for the stable installation of surface-bound metal complexes. The resulting surfaces are highly stabilized toward complex loss with excellent thermal, photochemical, and electrochemical stabilities. The procedure involves binding 3-azidopropyltrimethoxysilane (APTMS) to nanostructured mesoporous TiO2 or tin-doped indium oxide (ITO) electrodes by silane attachment followed by azide-terminated, Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reactions with an alkyne-derivatized ruthenium(II) polypyridyl complex. The chromophore-modified electrodes display enhanced photochemical and electrochemical stabilities compared to phosphonate surface binding with extended photoelectrochemical oxidation of hydroquinone for more than ∼6 h with no significant decay.

Enrichment of S-Palmitoylated Proteins for Mass Spectrometry Analysis.

As the 10-year anniversary of their first introduction approaches, alkynyl fatty acids have revolutionized the analysis of S-palmitoylation dynamics, acting as functional mimics incorporated into native modification sites in cultured cells. The alkyne functional group provides a robust handle for bioorthogonal Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to reporter-linked azides, forming a stable conjugate for enrichment for mass spectrometry analysis or in-gel fluorescence. Importantly, metabolic labeling enables time-dependent analysis of S-palmitoylation dynamics, which can be used to profile incorporation and turnover rates across the proteome. Here we present a protocol for cell labeling, click chemistry conjugation, enrichment, and isobaric tandem mass tag labeling for quantitative mass spectrometry analysis of protein S-palmitoylation.

Antimicrobial Titanium Surface via Click-Immobilization of Peptide and Its in Vitro/Vivo Activity.

The use of antimicrobial peptides (AMPs)-functionalized titanium implants is an efficient method for preventing bacterial infection. However, the attachment of AMPs to the surface of titanium implants remains a challenge. In this study, a "clickable" titanium surface was developed by using a silane coupling agent with an alkynyl group. The antimicrobial titanium implant was then constructed through the reaction between the "clickable" surface and azido-AMPs (PEG-HHC36:N3-PEG12-KRWWKWWRR) via click chemistry of Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC). Such an antimicrobial titanium implant, with an AMP density of 897.4 ± 67.3 ng/cm2 (2.5 ± 0.2 molecules per nm2) on the surface, exhibited good and stable antimicrobial activity, inhibited 90.2% of Staphylococcus aureus and 88.1% of Escherichia coli after 2.5 h of incubation, and even inhibited 69.5% of Staphylococcus aureus after 4 days of degradation. The CCK-8 assay indicated that the antimicrobial titanium implant exhibited negligible cytotoxicity to mouse bone mesenchymal stem cells. In vivo assay illustrated that this implant could kill 78.8% of Staphylococcus aureus after 7 days. This method has great potential for the preparation of antimicrobial titanium implants and the prevention of infections in the clinic.

Copper-catalyzed click reactions: quantification of retained copper using 64Cu-spiked Cu(I), exemplified for CuAAC reactions on liposomes

Abstract The Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) is a powerful, highly reliable and selective reaction which allows for a rapid synthesis in high yields and under mild conditions (pH, temperature). However, the cytotoxicity of copper requires its complete removal prior to an application in vivo. This is an issue especially when it comes to CuAAC reactions on macromolecular structures or drug delivery systems, as copper might be retained by these systems. Thus, a quantification of the final copper content of these systems is inevitable, which we exemplified for a CuAAC reaction on liposomes using 64Cu-spiked Cu(I). In this respect, a Cu(II) nitrate solution was irradiated at the TRIGA Mark II research reactor Mainz to obtain c.a. [64Cu]Cu(II). The irradiated solution was directly used for a CuAAC on liposomes. After purification, their copper content was calculated utilizing γ-ray spectrometry. Only 0.018% of the added 64Cu-activity was still present in the liposome containing fractions after purification. This refers to a total amount of copper of 0.17 ng. The amount of retained copper is so low, that an in vivo application of the liposomes is absolutely reasonable. Besides this particular study, the experimental methodology may be applied to study many other CuAAC reactions, used for the synthesis of radiolabeled or non-radioactive species, which are intended for human applications.

Reduction-responsive amphiphilic star copolymers with long-chain hyperbranched poly(ε-caprolactone) core and disulfide bonds for trigger release of anticancer drugs

In this contribution, the reduction-responsive star copolymers with long-chain hyperbranched poly(ε-caprolactone) (PCL) (HyperMacs) core and disulfide bonds were synthesized via Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. The HyperMacs core was constructed from disulfide-containing AB2-type PCL macromonomers, which possesses length-adjustable chain segments between branching points, large cavities, low degree of crystallinity, and reduction-responsivity. After grafted with poly(ethylene glycol), the reduction-responsive star copolymers can self-assemble into micelles in aqueous solution. The obtained micelles exhibited much lower critical micelle concentration (CMC) than their linear analogues. The reduction-responsivity from disulfide bonds makes them a promising carrier candidate for trigger release of anticancer drugs. The in vitro release results confirmed that their doxorubicin (DOX)-loaded micelles exhibited desirable reduction-triggered release performance. The cellular proliferation inhibition against HepG2 cells demonstrated that the DOX-loaded micelles showed a comparable anticancer activity with free DOX. Therefore, it can be expected that the reduction-sensitive micelles may serve as smart vehicles for intracellular delivery of anti-cancer drugs in tumour therapy.

Drug "Clicking" on Cell-Penetrating Fluorescent Nanoparticles for In Cellulo Chemical Proteomics.

Chemical proteomics approaches are widely used to identify molecular targets of existing or novel drugs. This manuscript describes the development of a straightforward approach to conjugate azide-labeled drugs via click chemistry to alkyne-tagged cell-penetrating fluorescent nanoparticles as a novel tool to study target engagement and/or identification inside living cells. A modification of the Baeyer test for alkynes allows monitoring the Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction, guaranteeing the presence of the drug on the solid support. As a proof of concept, the conjugation of the promiscuous kinase inhibitor dasatinib to Cy5-labeled nanoparticles is presented. Dasatinib-decorated fluorescent nanoparticles efficiently inhibited its protein target SRC in vitro, entered cancer cells, and colocalized with SRC in cellulo.

One-Pot Synthesis of pH/Redox Responsive Polymeric Prodrug and Fabrication of Shell Cross-Linked Prodrug Micelles for Antitumor Drug Transportation.

Shell cross-linked (SCL) polymeric prodrug micelles have the advantages of good blood circulation stability and high drug content. Herein, we report on a new kind of pH/redox responsive dynamic covalent SCL micelle, which was fabricated by self-assembly of a multifunctional polymeric prodrug. At first, a macroinitiator PBYP- ss- iBuBr was prepared via ring-opening polymerization (ROP), wherein PBYP represents poly[2-(but-3-yn-1-yloxy)-2-oxo-1,3,2-dioxaphospholane]. Subsequently, PBYP- hyd-DOX- ss-P(DMAEMA- co-FBEMA) prodrug was synthesized by a one-pot method with a combination of atom transfer radical polymerization (ATRP) and a Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction using a doxorubicin (DOX) derivative containing an azide group to react with the alkynyl group of the side chain in the PBYP block, while DMAEMA and FBEMA are the abbriviations of N, N-(2-dimethylamino)ethyl methacrylate and 2-(4-formylbenzoyloxy)ethyl methacrylate, respectively. The chemical structures of the polymer precursors and the prodrugs have been fully characterized. The SCL prodrug micelles were obtained by self-assembly of the prodrug and adding cross-linker dithiol bis(propanoic dihydrazide) (DTP). Compared with the shell un-cross-linked prodrug micelles, the SCL prodrug micelles can enhance the stability and prevent the drug from leaking in the body during blood circulation. The average size and morphology of the SCL prodrug micelles were measured by dynamic light scattering (DLS) and transmission electron microscopy (TEM), respectively. The SCL micelles can be dissociated under a moderately acidic and/or reductive microenvironment, that is, endosomal/lysosomal pH medium or high GSH level in the tumorous cytosol. The results of DOX release also confirmed that the SCL prodrug micelles possessed pH/reduction responsive properties. Cytotoxicity and cellular uptake analyses further revealed that the SCL prodrug micelles could be rapidly internalized into tumor cells through endocytosis and efficiently release DOX into the HeLa and HepG2 cells, which could efficiently inhibit the cell proliferation. This study provides a fast and precise synthesis method for preparing multifunctional polymer prodrugs, which hold great potential for optimal antitumor therapy.