Copper-catalyzed Azide-alkyne Cycloaddition Research Articles

Development of Novel Immobilized Copper-Ligand Complex for Click Chemistry of Biomolecules.

Copper-catalyzed azide-alkyne cycloaddition click (CuAAC) reaction is widely used to synthesize drug candidates and other biomolecule classes. Homogeneous catalysts, which consist of copper coordinated to a ligand framework, have been optimized for high yield and specificity of the CuAAC reaction, but CuAAC reaction with these catalysts requires the addition of a reducing agent and basic conditions, which can complicate some of the desired syntheses. Additionally, removing copper from the synthesized CuAAC-containing biomolecule is necessary for biological applications but inconvenient and requires additional purification steps. We describe here the design and synthesis of a PNN-type pincer ligand complex with copper (I) that stabilizes the copper (I) and, therefore, can act as a CuAAC catalyst without a reducing agent and base under physiologically relevant conditions. This complex was immobilized on two types of resin, and one of the immobilized catalyst forms worked well under aqueous physiological conditions. Minimal copper leaching was observed from the immobilized catalyst, which allowed its use in multiple reaction cycles without the addition of any reducing agent or base and without recharging with copper ion. The mechanism of the catalytic cycle was rationalized by density functional theory (DFT). This catalyst's utility was demonstrated by synthesizing coumarin derivatives of small molecules such as ferrocene and sugar.

Edge Functionalization of Bulk γ-Graphyne Facilitates Mechanical Exfoliation and Modulates the Mode of Sheet Stacking.

We have successfully achieved selective and efficient functionalization of sheet edges in microcrystalline multilayer γ-graphyne through two methods: cross-coupling with residual bromide edge groups and copper-catalyzed azide-alkyne cycloaddition (CuAAC) with edge terminal alkyne groups. This modification significantly enhances the ease of mechanical exfoliation and dispersibility of the sheets of γ-graphyne. Specifically, C18-grafted γ-graphyne forms stable dispersions in compatible organic solvents, allowing for the imaging of atomically thin layers of γ-graphyne for the first time. Additionally, we have discovered that phenylacetylide edge groups alter the preferred stacking mode of γ-graphyne sheets. Few-layer flakes of Ph-edge γ-graphyne exhibit a preference for the R3m space group, contrasting with the aperiodic stacking of Br-edge γ-graphyne. These results open the door for scalable exfoliation of few-layer flakes of γ-graphyne with a high aspect ratio, enabling potential applications in carbon electronics.

Synthesis of 1, 2, 3-triazole linked 5 fluorouracil - carbon dots -folate conjugates for target specific anticancer activity and cell imaging applications

This work provides a unique strategy to anchor the individual properties of fluorophores, pharmacophores and receptors in a single platform using a copper-catalyzed azide-alkyne cycloaddition reaction (CuAAC). Notably, this approach counters the long-term dispute associated with the target-specific drug delivery of 5 fluorouracil and its intracellular tracing. Significantly, the luminescence property of the carbon dots (CDs) and the anticancer activity of the 5 fluorouracil drug are well preserved, even after their structural modification. The resulting nano-hybrid conjugate shows good thermal stability, photo-stability and can selectively guide the drug molecule toward cancer cells and remain nontoxic to noncancerous (hFB) cells. The conjugation of folic acid to the nanohybrid surface promoted the folate receptor-facilitated endocytosis to the folate-positive (HeLa) cell lines over the folate-negative (MCF-7) cells, which enhanced cellular uptake and corresponding better cell apoptosis results. Around 18.2% of cell apoptosis (late + early) values were recorded for the folate-conjugated formulation compared to the folate-less formulation (12.3%) and pure 5 FU drug (7.9%) by a flow cytometry study. Cell cycle analysis confirmed that the populations of HeLa cells in the S phase were around 18.20% and 29.8% for the folate-less formulation and the folate-conjugated formulation, indicating all the formulations can hinder the DNA replication and thymidylate synthesis by introducing cell cycle arrest in the S phase, just like the pure 5 FU drug. Also, the location of the drug molecule can be simultaneously traced because of the luminescent nature of the CDs. Therefore, the developed system has potential in target-specific drug delivery and long-term drug molecule tracking applications.

Conjugated molecularly imprinted polymers based on covalent organic frameworks: Fluorescent sensing platform for specific capture of urea and elimination of ethyl carbamate

This study described the preparation of an azide covalent organic framework-embedded molecularly imprinted polymers (COFs(azide)@MIPs) platform for urea adsorption and indirect ethyl carbamate (EC) removal from Chinese yellow rice wine (Huangjiu). By modifying the pore surface of COFs using the copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction, COFs(azide) with a high fluorescence quantum yield and particular recognition ability were inventively produced. In order to selectively trap urea, the COFs(azide) were encased in an imprinted shell layer via imprinting technology. With a detection limit (LOD) of 0.016 μg L-1 (R2 = 0.9874), the COFs(azides)@MIPs demonstrated a good linear relationship with urea in the linear range of 0–5 μg L-1. Using real Huangjiu samples, the spiking recovery trials showed the viability of this sensing platform with recoveries ranging from 88.44 % to 109.26 % and an RSD of less than 3.40 %. The Huangjiu processing model system achieved 38.93 % EC reduction by COFs(azides)@MIPs. This research will open up new avenues for the treatment of health problems associated with fermented alcoholic beverages, particularly Huangjiu, while also capturing and removing hazards coming from food.

A 66-Nuclear All-Alkynyl Protected Peanut-Shaped Silver(I)/Copper(I) Heterometallic Nanocluster: Intermediate in Copper-Catalyzed Alkyne-Azide Cycloaddition.

Ligand-protected heterometallic nanoclusters in contrast to homo-metal counterparts show more broad applications due to the synergistic effect of hetero-metals but their controllable syntheses remain a challenge. Among heterometallic nanoclusters, monovalent Ag-Cu compounds are rarely explored due to much difference of Ag(I) and Cu(I) such as atom radius, coordination habits, and redox potential. Encouraged by copper-catalyzed alkyne-azide cycloaddition (CuAAC) reaction, comproportionation reaction of Cu(II)X2 and Cu(0) in the presence of (PhC≡CAg)n complex and molybdate generated a core-shell peanut-shaped 66-nuclear Ag(I)-Cu(I) heterometallic nanocluster, [(Mo4O16)2@Cu12Ag54(PhC≡C)50] (referred to as Ag54Cu12). The structure and composition of Ag-Cu heterometallic nanocluster are fully characterized. X-ray single crystal diffraction reveals that Ag54Cu12 has a peanut-shaped silver(I)/copper(I) heterometallic nanocage protected by fifty phenylacetylene ligands in µ3-modes and encapsulated two mutually twisted tetramolybdates. Heterometallic nanocage contains a 54-Ag-atom outer ellipsoid silver cage decorated by 12 copper inside wall. Nanosized Ag54Cu12 is a n-type narrow-band-gap semiconductor with a good photocurrent response. Preliminary experiments demonstrates that Ag54Cu12 itself and activated carbon supported Ag54Cu12/C are effective catalysts for 1,3-dipole cycloaddition between alkynes and azides at ambient conditions. The work provides not only a new synthetic route toward Ag(I)-Cu(I) nanoclusters but also an important heterometallic intermediate in CuAAC catalytic reaction.

Synthesis of block macro-CTAs by HNTs@Cu-catalyzed click reaction: Suitable CTAs for synthesizing multiblock polymers via RAFT polymerization

Multiblock copolymer macrochain-transfer agents (MBP-CTAs) can be used to synthesize MBPs in one pot via reversible addition-fragmentation chain-transfer (RAFT) polymerization. MBP-CTAs can be efficiently synthesized via a copper-catalyzed azide-alkyne cycloaddition reaction. However, traditional purification methods cannot sufficiently purify MBP-CTAs, resulting in the failure of RAFT. In this study, it was discovered that using a supported catalytic system, namely, Cu@A-HNTs, inhibited the connection between Cu and S. The residual Cu content in the MBP-CTA was successfully reduced to a low level (7.88 µg/mg). Then, hydrophilic and hydrophobic MBPs were synthesized through RAFT polymerization using the MBP-CTAs. Subsequently, multiblock copoly(methyl methacrylate-b-butyl methacrylate-CTA)s (PMMA-b-PBMA-c-CTA)s were successfully prepared. The MBP-CTAs and MBPs were characterized using ultraviolet–visible spectroscopy, 1H nuclear magnetic resonance, size-exclusion chromatography, matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry, differential scanning calorimetry measurements and uniaxial tensile tests. The results of performance testing of PMMA-b-PBMA-c-CTAs demonstrated that their fracture stress and strain significantly increased comparing with that of PMMA-b-CTAs.

Selective Duplex Formation in Mixed Sequence Libraries of Synthetic Polymers.

Recognition-encoded melamine oligomers (REMO) are synthetic polymers that feature an alternating 1,3,5-triazine-piperazine backbone and side-chains equipped with either a phenol or phosphine oxide recognition unit. An automated method for the solid-phase synthesis (SPS) of REMO of any specified sequence has been developed starting from dichlorotriazine monomer building blocks. Complementary homo-oligomers with either six phenols or six phosphine oxides were synthesized and shown to form a stable duplex in nonpolar solvents by NMR denaturation experiments. The duplex was covalently trapped by equipping the ends of the oligomers with an azide and an alkyne group and using a copper-catalyzed alkyne-azide cycloaddition (CuAAC) reaction. The SPS methodology was adapted to synthesize mixed sequence libraries by using a mixture of two different dichlorotriazine building blocks in each coupling cycle of an oligomer synthesis. The resulting libraries contain statistical mixtures of all possible sequences. The self-assembly properties of these libraries were screened by using the CuAAC reaction to trap any duplexes present. In mixed sequence libraries of 6-mers, the trapping experiments showed that only sequence-complementary oligomers formed duplexes at micromolar concentrations in dichloromethane. The automated synthesis approach developed here provides access to large libraries of mixed sequence synthetic polymers, and the covalent trapping experiment provides a convenient tool for screening functional properties of mixtures. The results suggest high-fidelity sequence-selective duplex formation in mixtures of 6-mer sequences of the REMO architecture.

Metal-Free Click-Chemistry: A Powerful Tool for Fabricating Hydrogels for Biomedical Applications.

Increasing interest in the utilization of hydrogels in various areas of biomedical sciences ranging from biosensing and drug delivery to tissue engineering has necessitated the synthesis of these materials using efficient and benign chemical transformations. In this regard, the advent of "click" chemistry revolutionized the design of hydrogels and a range of efficient reactions was utilized to obtain hydrogels with increased control over their physicochemical properties. The ability to apply the "click" chemistry paradigm to both synthetic and natural polymers as hydrogel precursors further expanded the utility of this chemistry in network formation. In particular, the ability to integrate clickable handles at predetermined locations in polymeric components enables the formation of well-defined networks. Although, in the early years of "click" chemistry, the copper-catalyzed azide-alkyne cycloaddition was widely employed, recent years have focused on the use of metal-free "click" transformations, since residual metal impurities may interfere with or compromise the biological function of such materials. Furthermore, many of the non-metal-catalyzed "click" transformations enable the fabrication of injectable hydrogels, as well as the fabrication of microstructured gels using spatial and temporal control. This review article summarizes the recent advances in the fabrication of hydrogels using various metal-free "click" reactions and highlights the applications of thus obtained materials. One could envision that the use of these versatile metal-free "click" reactions would continue to revolutionize the design of functional hydrogels geared to address unmet needs in biomedical sciences.

Abstract 7142: Optimizing estradiol dimers through linker design: Enhancing anticancer efficacy by targeting microtubule dynamics

Abstract We investigated the development and anticancer efficacy of novel estradiol dimers (ED), targeting the dynamics of microtubules—an essential factor in effective cancer therapy. Employing copper-catalyzed azide-alkyne cycloaddition (CuAAC), two distinct series of ED variants were synthesized. The initial series included thirteen dimers, derived from 17α-ethinyl estradiol, incorporating varied five-atom linkers with central carbon, nitrogen, or oxygen atoms, and, in some instances, modified with benzyl groups containing hydroxy or methoxy substituents. These dimers were evaluated for their cytotoxic properties and impact on the cell cycle, revealing certain structures with substantial efficacy, compared with the original ED. The subsequent series, encompassing twelve variants, concentrated on structural alterations within the aromatic bridges linking the estradiol units. In this series, linkers of simpler composition and fewer substituents demonstrated increased selectivity and potency against cancer cell lines, particularly ED3 and ED5, which exhibited cytotoxicity levels comparable to established tubulin inhibitors. Both dimers series showed the ability to disrupt microtubule assembly and mitotic processes, confirmed through cell-based evaluations and in vitro tubulin assembly assays. Complementing these results, in silico modelling and binding free energy calculations exhibited a strong correlation with the demonstrated biological activities, especially in terms of binding affinity to the colchicine site. Collectively, these findings highlight the promise of estradiol dimers as potential anticancer agents and emphasize the critical role of linker structures in enhancing their biological effectiveness. Work was supported by the Ministry of Education, Youth and Sports of the Czech Republic through infrastructural projects (CZ-OPENSCREEN - LM2018130; EATRIS-CZ - LM2018133) and National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102) - Funded by the European Union - Next Generation EU. Citation Format: Petr Dzubak, Michal Jurášek, Jiří Řehulka, Soňa Gurská, Pavel Polishchuk, Pavel Dráber, Lukáš Huml, Pavel B. Drašar, Alexandra Ivanova, Olena Mokshyna, Lenka Hrubá, Marián Hajdúch. Optimizing estradiol dimers through linker design: Enhancing anticancer efficacy by targeting microtubule dynamics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7142.

Semisynthesis and characterization of versatile azide intermediates using sodium alginate and its homopolymeric derivatives as starting material

Alginate, a polyuronic biopolymer composed of mannuronic and guluronic acid units, contain hydroxyl and carboxyl groups as targeting modification sites to obtain structures with new and/or improved biological properties. The copper-catalyzed azide-alkyne cycloaddition (CuAAC) is a versatile click reaction for polymer functionalization, but it typically requires a “pre-click” modification to introduce azide or alkyne groups. Here, we described a straightforward chemical path to selectively modify alginate carboxyl groups producing versatile azido derivatives through N-acylation using 3-azydopropylamine. The resulting azide-functionalized polysaccharides underwent click chemistry to yield amino derivatives, confirmed by NMR and FTIR analyses. The 1H NMR spectrum reveals a characteristic triazole group signal at 8.15 ppm. The absence of the azide FTIR band for all amino derivatives, previously observed for the N-acylation products, indicated reaction success. Antibacterial and antioxidant assessments revealed that the initial polysaccharide lacks E. coli inhibition, while the click chemistry-derived amine products exhibit growth inhibition at 5.0 mg/mL. Lower molecular weight derivatives demonstrate superior DPPH scavenging ability, particularly amino-derivatives (24–33 % at 1.2 mg/mL). This innovative chemical pathway offers a promising strategy for developing polysaccharide structures with enhanced properties, demonstrating potential applications in various fields.

Minimalist Tetrazine N-Acetyl Muramic Acid Probes for Rapid and Efficient Labeling of Commensal and Pathogenic Peptidoglycans in Living Bacterial Culture and During Macrophage Invasion.

N-Acetyl muramic acid (NAM) probes containing alkyne or azide groups are commonly used to investigate aspects of cell wall synthesis because of their small size and ability to incorporate into bacterial peptidoglycan (PG). However, copper-catalyzed alkyne-azide cycloaddition (CuAAC) reactions are not compatible with live cells, and strain-promoted alkyne-azide cycloaddition (SPAAC) reaction rates are modest and, therefore, not as desirable for tracking the temporal alterations of bacterial cell growth, remodeling, and division. Alternatively, the tetrazine-trans-cyclooctene ligation (Tz-TCO), which is the fastest known bioorthogonal reaction and not cytotoxic, allows for rapid live-cell labeling of PG at biologically relevant time scales and concentrations. Previous work to increase reaction kinetics on the PG surface by using tetrazine probes was limited because of low incorporation of the probe. Described here are new approaches to construct a minimalist tetrazine (Tz)-NAM probe utilizing recent advancements in asymmetric tetrazine synthesis. This minimalist Tz-NAM probe was successfully incorporated into pathogenic and commensal bacterial PG where fixed and rapid live-cell, no-wash labeling was successful in both free bacterial cultures and in coculture with human macrophages. Overall, this probe allows for expeditious labeling of bacterial PG, thereby making it an exceptional tool for monitoring PG biosynthesis for the development of new antibiotic screens. The versatility and selectivity of this probe will allow for real-time interrogation of the interactions of bacterial pathogens in a human host and will serve a broader utility for studying glycans in multiple complex biological systems.

Aptamer-Targeted Dendrimersomes Assembled from Azido-Modified Janus Dendrimers "Clicked" to DNA.

Amphiphilic Janus dendrimers (JDs), synthetic alternatives to lipids, have the potential to expand the scope of nanocarrier delivery systems. JDs self-assemble into vesicles called dendrimersomes, encapsulate both hydrophobic cargo and nucleic acids, and demonstrate enhanced stability in comparison to lipid nanoparticles (LNPs). Here, we report the ability to enhance the cellular uptake of Janus dendrimersomes using DNA aptamers. Azido-modified JDs were synthesized and conjugated to alkyne-modified DNAs using copper-catalyzed azide alkyne cycloaddition. DNA-functionalized JDs form nanometer-sized dendrimersomes in aqueous solution via thin film hydration. These vesicles, now displaying short DNAs, are then hybridized to transferrin receptor binding DNA aptamers. Aptamer-targeted dendrimersomes show improved cellular uptake as compared to control vesicles via fluorescence microscopy and flow cytometry. This work demonstrates the versatility of using click chemistry to conjugate functionalized JDs with biologically relevant molecules and the feasibility of targeting DNA-modified dendrimersomes for drug delivery applications.

Glycomimetic inhibitors of tandem-repeat galectins: Simple and efficient

The binding of human galectins by glycomimetic inhibitors is a promising therapeutic approach. The structurally distinct group of tandem-repeat galectins has scarcely been studied so far, and there is hardly any knowledge on their ligand specificity or their inhibitory potential, particularly concerning non-natural carbohydrates. Here, we present the synthesis of a library of seven 3-O-disubstituted thiodigalactoside-derived glycomimetics and their affinity to two tandem-repeat galectins, Gal-8 and Gal-9. The straightforward synthesis of these glycomimetics involved dibutyltin oxide-catalyzed 3,3́-O-disubstitution of commercially available unprotected thiodigalactoside, and conjugation of various aryl substituents by copper-catalyzed Huisgen azide-alkyne cycloaddition (CuAAC). The inhibitory potential of the prepared glycomimetics for Gal-8 and Gal-9 was assessed, and compared with the established galectins Gal-1 and Gal-3. The introduction of C-3 substituents resulted in an over 40-fold increase in affinity compared with unmodified TDG. The structure-affinity relations within the studied series were discussed using molecular modeling. Furthermore, the prepared glycomimetics were shown to scavenge Gal-8 and Gal-9 from the surface of cancer cells. This pioneering study on the synthetic inhibitors especially of Gal-9 identified lead compounds that may be used in further biomedical research.

MOFs Modulate Copper Trafficking in Tumor Cells for Bioorthogonal Therapy.

In situ drug synthesis using the copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction has attracted considerable attention in tumor therapy because of its satisfactory effectiveness and reduced side-effects. However, the exogenous addition of copper catalysts can cause cytotoxicity and has hampered biomedical applications in vivo. Here, we design and synthesize a metal-organic framework (MOF) to mimic copper chaperone, which can selectively modulate copper trafficking for bioorthogonal synthesis with no need of exogenous addition of copper catalysts. Like copper chaperones, the prepared ZIF-8 copper chaperone mimics specifically bind copper ions through the formation of coordination bonds. Moreover, the copper is unloaded under the acidic environment due to the dissipation of the coordination interactions between metal ions and ligands. In this way, the cancer cell-targeted copper chaperone mimics can selectively transport copper ions into cells. Regulation of intracellular copper trafficking may inspire constructing bioorthogonal catalysis system with reduced metal cytotoxicity in live cells.

On the mechanochemical synthesis of C-scorpionates with an oxime moiety and their application in the copper-catalyzed azide–alkyne cycloaddition (CuAAC) reaction

Integrating the principles of green chemistry to develop sustainable processes.

Iterative click reactions using trivalent platforms for sequential molecular assembly.

A facile synthesis of multi(triazole)s by iterative click reactions is disclosed. Good functional group tolerance of sequential click assembly by sulfur-fluoride exchange (SuFEx), copper-catalyzed azide-alkyne cycloaddition (CuAAC), and thia-Michael reaction realizes the iterative click reactions. Diverse multi(triazole)-type mid-molecules can be synthesized easily from readily available modules through good chemoselective reactions without functional group transformation steps.

Real-time Visualization of Transcribed mRNA via Click Chemistry in a Liposomal Space.

We present a CuAAC (Copper-Catalyzed Azide-Alkyne Cycloaddition) reaction protocol designed for the visualization of mRNA. To achieve this, we synthesized stable mRNA molecules incorporating the modified nucleoside analog, EU, a crucial element for fluorophore attachment. Leveraging this modified mRNA, we successfully executed the CuAAC reaction, wherein the pro-fluorophore, coumarin, was conjugated to EU on the mRNA through our meticulously designed CuAAC process. This innovative approach resulted in the emission of fluorescence, enabling both precise quantification and visual observation of mRNA. Furthermore, we demonstrated the feasibility of concurrent mRNA synthesis and visualization by seamlessly integrating the CuAAC reaction mix into the mRNA transcription process. Additionally, our novel methodology opens avenues for prospective real-time monitoring of mRNA transcription within artificial cells. These advancements hold significant promise for expanding our comprehension of fundamental cellular processes and finding applications across diverse biological contexts in the future.

Coumarin-azasugar-benzyl conjugates as non-neurotoxic dual inhibitors of butyrylcholinesterase and cancer cell growth.

We have applied the copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction to prepare a library of ten coumarin-azasugar-benzyl conjugates and two phthalimide-azasugar-benzyl conjugates with potential anti-Alzheimer and anti-cancer properties. The compounds were evaluated as cholinesterase inhibitors, demonstrating a general preference, of up to 676-fold, for the inhibition of butyrylcholinesterase (BuChE) over acetylcholinesterase (AChE). Nine of the compounds behaved as stronger BuChE inhibitors than galantamine, one of the few drugs in clinical use against Alzheimer's disease. The most potent BuChE inhibitor (IC50 = 74 nM) was found to exhibit dual activities, as it also showed high activity (GI50 = 5.6 ± 1.1 μM) for inhibiting the growth of WiDr (colon cancer cells). In vitro studies on this dual-activity compound on Cerebellar Granule Neurons (CGNs) demonstrated that it displays no neurotoxicity.

Novel isatin-triazole based thiosemicarbazones as potential anticancer agents: synthesis, DFT and molecular docking studies.

Thiosemicarbazones of isatin have been found to exhibit versatile bioactivities. In this study, two distinct types of isatin-triazole hybrids 3a and 3b were accessed via copper-catalyzed azide-alkyne cycloaddition reaction (CuAAC), together with their mono and bis-thiosemicarbazone derivatives 4a-h and 5a-h. In addition to the characterization by physical, spectral and analytical data, a DFT study was carried out to obtain the optimized geometries of all thiosemicarbazones. The global reactivity values showed that among the synthesized derivatives, 4c, 4g and 5c having nitro substituents are the most soft compounds, with compound 5c having the highest electronegativity and electrophilicity index values among the synthesized series, thus possessing strong binding ability with biomolecules. Molecular docking studies were performed to explore the inhibitory ability of the selected compounds against the active sites of the anticancer protein of phosphoinositide 3-kinase (PI3K). Among the synthesized derivatives, 4-nitro substituted bisthiosemicarbazone 5c showed the highest binding energy of -10.3 kcal mol-1. These findings demonstrated that compound 5c could be used as a favored anticancer scaffold via the mechanism of inhibition against the PI3K signaling pathways.

Highly efficient functionalization of hydroxypropyl cellulose with glucuronic acid through click chemistry under microwave irradiation as potential biomaterial with therapeutic properties

A straightforward and effective method to chemically conjugate hydroxypropyl cellulose (HPC) to GlcA is described via microwave assisted copper-catalyzed azide-alkyne cycloaddition reaction (CuAAC) between the azide functionalized HPC and the alkynyl functionalized GlcA, reducing reaction times from 24 h to 2 h. Two types of HPC azide derivatives, including an ester (ES) or a carbamate (CB) functional group, were synthesized and characterized by FT-IR and NMR spectroscopy that revealed the efficiency in the incorporation of the azide moieties in the cellulose backbone. These compounds were then conjugated with the alkynyl GlcA by CuAAC, affording the desired glycopolymers, HPA-ES-GlcA and HPA-CB-GlcA in excellent yields. The whole process involved rapid and effective reactions as well as easy purification techniques, using mostly non-toxic solvents, contributing to the sustainability of the synthesis These glycopolymers were characterized by FT-IR, NMR, TGA, DTA and SEM. Furthermore, the cytotoxicity, anti-inflammatory and antibacterial properties of the new glycopolymers were tested and it was found that these new HPC-GlcA conjugates presented low cytotoxicity, good anti-inflammatory and antibacterial properties. Overall, our results showed that HPC-GlcA polymers are suitable as potential GAG mimics for biomedical applications, therefore further investigations will be carried out.