Bioconjugate Chemistry最新文献

{"title":"Design and Synthesis of an Adamantane Phosphoramidite for Programmable and Automated Oligonucleotide-Functionalization.","authors":"Yuxiang Cong, Xiaoxing Chen, Thulasiram Bathini, Gang Chen, Da Han, Yangen Huang, Ruowen Wang","doi":"10.1021/acs.bioconjchem.5c00163","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00163","url":null,"abstract":"<p><p>Solid-phase synthesis has revolutionized the programmable preparation of oligonucleotides (ONs), enabling precise gene expression modulation and expanding their applications in therapeutic and material sciences. To further enhance ON functionality, this study introduces a novel adamantane-based phosphoramidite for oligonucleotide modification. Adamantane, known for its hydrophobicity and stability, was incorporated into nucleic acid aptamers using automated synthesis. Two aptamers─Sgc8 and AS1411─were functionalized with one or two adamantane units, and the products were purified and validated using high-performance liquid chromatography and mass spectrometry. The incorporation of adamantane significantly altered the aptamers' polarity and facilitated their self-assembly with poly-β-cyclodextrin, forming stable supramolecular complexes, as demonstrated by polyacrylamide gel electrophoresis. Additionally, adamantane-modified AS1411 exhibited enhanced degradation of its target protein, nucleolin, in MCF-7 cells, suggesting potential utility in targeted protein regulation. These findings establish a versatile platform for functionalizing ONs, broadening their potential for biomedical and nanotechnological applications.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2025-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144118372","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Conceptualization and Preliminary Characterization of Poloxamer-Based Hydrogels for Biomedical Applications.","authors":"Marta Tuszynska, Joanna Skopinska-Wisniewska, Mateusz Bartniak, Anna Bajek","doi":"10.1021/acs.bioconjchem.5c00030","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00030","url":null,"abstract":"<p><p>Poloxamers are widely used in biomedical applications, but their effectiveness depends on achieving an optimal sol-gel phase transition near body temperature. This study evaluates three different poloxamer mixtures for their potential in treating meniscus tears, focusing on gel formation, injectability, and cell compatibility. The rheological properties, cytotoxicity assessments, and cellular migration experiments were studied using NIH/3T3 fibroblast cells as the standard experimental model for primary research. The poloxamer hydrogels showed properties well suited to injectable or drug delivery systems. Specifically, the combination of Synperonic F108 and Poloxamer 188 tended to show less adhesion and more aggregation, followed by a greater number of viable cells, suggesting its utility as a coating or foundational matrix. Concurrently, the Kolliphor 407 and Poloxamer 188 combination exhibited increased viscosity, maintaining a gel state at physiological temperature. Its biocompatibility indicated the potential for injectable controlled-release systems for musculoskeletal injuries. Our findings demonstrate that the poloxamer concentration and composition significantly influence their biomedical applications. These triblock copolymer systems indicated useful characteristics for surgical applications, such as favorable sol-gel transition kinetics and biocompatibility, suggesting potential applications in osteoarticular regeneration.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2025-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144092009","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Employing the SpyTag-SpyCatcher Reaction for the Modification of Supramolecular Polymers with Functional Proteins.","authors":"Fenna W B Craenmehr, Alexander Gräwe, Victor A Veenbrink, Riccardo Bellan, Maarten Merkx, Patricia Y W Dankers","doi":"10.1021/acs.bioconjchem.5c00046","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00046","url":null,"abstract":"<p><p>Supramolecular assemblies hold great potential as biomaterials for several biomedical applications. The modification of supramolecular biomaterials is needed to achieve controlled bioactive functions. Supramolecular ureidopyrimidinone (UPy) monomers have been shown to assemble into long supramolecular polymers that can be functionalized with bioactive peptides and visualized as UPy-fibers. So far, the introduction of biological functionality has been limited to small molecules and peptides. Here, we describe a general method based on SpyTag-SpyCatcher chemistry for conjugating full-length proteins with biologically relevant functions to μm-long UPy fibers via native peptide bond formation, yielding 100% conversion in a 5:95 mol % coassembly of UPy-SpyTag with UPy-glycinamide. The conjugation of monoclonal antibodies is performed using photo-cross-linkable protein G domains. We demonstrate intact fibers and colocalization of antibodies and UPy-fibers using biophysical and imaging methods and achieve recruitment of supramolecular assemblies to the surface of mammalian cells via the EGFR-specific antibody Cetuximab. The approach introduced here represents a robust and widely applicable postassembly modification method that shows promise in the functionalization of future biomaterials.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143952197","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a Multi-Stimuli-Responsive Magnetic Nanogel-Hydrogel Nanocomposite for Prolonged and Controlled Doxorubicin Release.","authors":"Ghasem Rezanejade Bardajee, Hossein Mahmoodian, Negin Shafiei, Bita Amiri","doi":"10.1021/acs.bioconjchem.5c00083","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00083","url":null,"abstract":"<p><p>The development of advanced drug delivery systems that offer precise, controlled, and sustained release of therapeutic agents remains a significant challenge, particularly for applications in oncology where effective targeting and prolonged drug exposure are essential. We synthesized and characterized a multistimuli-responsive magnetic nanogel-hydrogel nanocomposite (MNHNC) designed for controlled and extended drug release, with an emphasis on anticancer drug delivery. The MNHNC was developed by incorporating poly(<i>N</i>-isopropylacrylamide-<i>co</i>-acrylamide) (p(NIPAM-<i>co</i>-AAm)) nanogels (NGs) within a net-shaped salep-grafted poly(acrylic acid) (PAA) hydrogel matrix, coupled with in situ formation of Fe₃O₄ nanoparticles to introduce magnetic responsiveness and serve as a cross-linking agent. The nanocomposite exhibited notable swelling capabilities, achieving equilibrium values of 706 g/g at pH 9 (25 °C) and 603 g/g at physiological temperature (37 °C, pH 7.4). Additionally, MNHNC demonstrated responsiveness to pH, temperature, and magnetic fields, facilitating controlled drug release. Using doxorubicin (DOX) as a model drug, MNHNC exhibited dual pH sensitivity (NG at pH 5.4 and MNHNC at pH 7.4) and achieved a prolonged release profile of 400 h, significantly surpassing conventional systems, including our previous nanocomposite. Release kinetics followed a super case-II transport mechanism, where swelling primarily governed drug diffusion. Furthermore, the application of a magnetic field enabled fine-tuning of the release rate, offering an additional layer of control. The kinetic study indicated that the drug release from MNHNC followed zero-order kinetics under certain conditions, ensuring a consistent release rate over time, which is highly desirable for maintaining therapeutic efficacy. The Korsmeyer-Peppas model further confirmed the super case-II transport mechanism, highlighting the significant influence of polymer relaxation and swelling on the release process. The Hixson-Crowell model also demonstrated the role of matrix erosion in the drug release mechanism. The results showed a marked improvement in pH and temperature sensitivity compared to previous formulations, enhanced mechanical stability due to the integration of Fe₃O₄ nanoparticles, and the ability to modulate drug release through external magnetic fields. In vitro cytotoxicity assessment using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay demonstrated the biocompatibility of the MNHNC, with over 95% cell viability in the absence of DOX, confirming its nontoxic nature. Upon DOX loading, MNHNC exhibited a proper anticancer effect against cancer cell lines, showing a dose-dependent reduction in cell viability. The robust mechanical stability, biocompatibility, and multistimuli responsiveness of MNHNC position it as a promising candidate for advanced, targeted drug delivery systems.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144074911","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Design, Synthesis, and Biological Evaluation of a Novel Long-Acting Human Complement C3 Inhibitor Synthesized via the PASylation-Lipidation Modular (PLM) Platform.","authors":"Chengcheng Wang, Yapeng Wang, Yu Duan, Yuanzhen Dong, Haoju Hua, Huixin Cui, Shuaiyi Huang, Zongqing Huang, Jianguang Lu, Chunyong Ding, Zhengyan Cai, Jun Feng","doi":"10.1021/acs.bioconjchem.5c00108","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00108","url":null,"abstract":"<p><p>The complement system is essential for immune defense, but its dysregulation contributes to various complement-mediated disorders, including paroxysmal nocturnal hemoglobinuria (PNH). <b>CP40</b> (a cyclic peptide also known as AMY101), effectively inhibits complement activation by preventing the initial binding of the C3 substrate to convertase. Despite its potency, <b>CP40</b> has a very short plasma half-life when unbound to human C3, necessitating frequent dosing. We developed a novel PASylation-Lipidation Modular (PLM) platform. This platform incorporates a solubilizing PAS module and a half-life-extending lipid moiety into <b>CP40</b> via a chemical linker. Systematic optimization of the spacer and lipid components in PLM-modified <b>CP40</b> analogues identified <b>6C1</b> as a lead compound. Compared to <b>CP40</b>, <b>6C1</b> exhibited a 5-fold increase in antihemolytic potency in the classical complement pathway and a 6.3-fold improvement in solubility. <i>In vivo</i> studies demonstrated that PLM-<b>CP40</b> analogues possess superior pharmacokinetic properties, with a 15.6-fold extension in half-life relative to unmodified <b>CP40</b>. Mechanistic studies revealed that the PLM platform extends half-life by interacting with albumin, which serves as a circulating depot for the compound. Surface plasmon resonance analysis and hemolysis assays postalbumin incubation demonstrated that PLM modifications maintain receptor affinity by strategically positioning the albumin-binding moiety away from the peptide region, preserving its biological activity. In a clinically relevant <i>in vitro</i> model of complement-mediated hemolysis in PNH, <b>6C1</b> effectively reduced erythrocyte lysis. The PLM platform thus offers a versatile strategy for enhancing peptide therapeutics by improving solubility, extending circulation time, and increasing efficacy, broadening their therapeutic potential.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143951384","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transferrin Receptor-Targeted Aptamer-Drug Conjugate Overcomes Blood-Brain Barrier for Potent Glioblastoma Therapy.","authors":"Xinyue Zhao, Jiaxuan He, Yingda Chen, Jianpei Zheng, Xuefeng Li, Ting Fu, Sitao Xie, Xiangsheng Liu, Weihong Tan","doi":"10.1021/acs.bioconjchem.5c00137","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00137","url":null,"abstract":"<p><p>Glioblastoma (GBM) is the leading primary malignant tumor in the central nervous system. Current clinical therapeutics for treating GBM patients yield limited benefits. However, the development of new therapeutic methods is hindered because the blood-brain barrier (BBB) restricts drug penetration. The transferrin receptor (TfR) is highly expressed by brain endothelial cells and GBM cells, and it is considered a promising target for GBM drug delivery. Here, we modularly constructed a TfR-targeted aptamerdrug conjugate (ApDC) by linking a TfR aptamer (HG1-9) and a highly potent anti-tubulin drug, monomethyl auristatin E (MMAE), to cross the BBB and deliver GBM treatment. The targeting and BBB transport abilities of the TfR-targeted ApDC (HG1-9-MMAE) for GBM were evaluated in cultured vascular endothelial bEnd.3 cells and human GBM U-87 MG Luc2 cells, together with an <i>in vitro</i> transwell BBB model. Potent antitumor effects of HG1-9-MMAE were demonstrated by <i>in vitro</i> cellular proliferation assays and <i>in vivo</i> tumor inhibition in both subcutaneous and orthotopic GBM models. Our findings indicated that ApDC could be an efficient drug delivery strategy to treat GBM.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143954180","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Positron Emission Tomography and Optical Imaging to Monitor Bioorthogonal Diels–Alder Click Chemistry of Trastuzumab with a Porphyrin","authors":"Sara R. D. Gamelas,&nbsp;Shayla Shmuel,&nbsp;Cristina Simó,&nbsp;Alex Vanover,&nbsp;João P. C. Tomé,&nbsp;Augusto C. Tomé,&nbsp;Leandro M. O. Lourenço* and Patrícia M. R. Pereira*,&nbsp;","doi":"10.1021/acs.bioconjchem.5c0005610.1021/acs.bioconjchem.5c00056","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00056https://doi.org/10.1021/acs.bioconjchem.5c00056","url":null,"abstract":"<p >Click chemistry to allow <i>in vivo</i> conjugation of a fluorophore porphyrin (Por)–tetrazine (Tz) with the human epidermal growth factor receptor 2 (HER2)–targeting trastuzumab conjugated with trans-cyclooctene (TCO) is described here. <i>In vitro</i> experiments confirmed successful click reactions between Por–Tz and trastuzumab–TCO and validated preserved trastuzumab immunoreactivity (no significant change in HER2 binding, <i>p</i> &gt; 0.05). Positron emission tomography (PET) of [⁸⁹Zr]Zr–DFO-trastuzumab–TCO demonstrated 17.1 ± 2.9% injected dose per gram of tumor at 48 h postinjection. Optical imaging showed an ∼10-fold increase in the click group for Por–Tz when compared with Por–Tz alone. This preclinical data demonstrate a pretargeted approach for dual PET and optical imaging of HER2-expressing tumors.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 5","pages":"1013–1020 1013–1020"},"PeriodicalIF":4.0,"publicationDate":"2025-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144098046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chelating [²²⁷Th]Th⁴⁺ for Peptide Receptor Radionuclide Therapy of Neuroendocrine Tumors.","authors":"Luke Wharton, Scott W McNeil, Harrison Meeres, Diduo Zhang, Aidan Ingham, Helen Merkens, Maryam Osooly, Cristina Rodríguez-Rodríguez, François Bénard, Hua Yang","doi":"10.1021/acs.bioconjchem.5c00129","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00129","url":null,"abstract":"<p><p>Targeted alpha therapy (TAT) has shown high promise for the effective treatment of advanced stage cancers. Of the proposed radionuclides for TAT, Thorium-227 represents an interesting candidate given its relatively long half-life, 18.7 days, and the cascade of short-lived, high-potency, alpha-emitting daughter progeny in its decay scheme. However, to date few chelators exist which can effectively and stably bind [²²⁷Th]Th⁴⁺ at molar activities high enough for TAT. To address this challenge, this study investigated various chelating ligands for coordination of [²²⁷Th]Th⁴⁺. H₄noneunpaX was identified as a promising chelator, demonstrating radiolabeling with [²²⁷Th]Th⁴⁺ at concentrations of 10^-6 M (A_m = 272 kBq/nmol). The coordination characteristics of [Th(noneunpaX)] have been investigated through ¹H NMR spectroscopy, mass spectrometry, and DFT calculations. In this study, we also investigate for the first time the pairing of Th-227 with a peptide-based bioconjugate and evaluate the <i>in vivo</i> biodistribution characteristics. [²²⁷Th]Th-nonenupaX-Ahx-Tyr³-TATE was prepared under mild conditions (ambient temperature, 30 min) and evaluated in NRG mice bearing AR42J xenografts as a model for pancreatic neuroendocrine tumors. The ²²⁷Th-labeled radiopeptide showed high uptake in tumors (25.8±6.2 %IA/g at 3 h p.i.) and low uptake in non-targeted organs. Although some release of Th-227 was noted in serum stability studies this was not observed <i>in vivo</i>. This ligand architecture serves as an interesting framework for future optimization, which will involve improvements to the overall stability by enhancing the rigidity of the backbone and assessing other pendent donor groups with a stronger affinity toward [²²⁷Th]Th⁴⁺. Overall, this study demonstrated for the first time the viability of using peptide-based targeting to effectively deliver Th-227 to tumor sites.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":""},"PeriodicalIF":4.0,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143951307","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Carrier Free 1,2,3,4,6-O-Pentagalloylglucose Nanoparticles for Treatment of Acute Lung Injury","authors":"Qi Zhang,&nbsp;Ying Wang,&nbsp;Zixuan Yang,&nbsp;Zhiming Xin,&nbsp;Haohua Deng* and Wei Chen*,&nbsp;","doi":"10.1021/acs.bioconjchem.5c0019710.1021/acs.bioconjchem.5c00197","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00197https://doi.org/10.1021/acs.bioconjchem.5c00197","url":null,"abstract":"<p >Acute lung injury (ALI) is a severe lung disease with a high mortality rate, and novel therapeutic strategies must be developed crucially. The amelioration of inflammation and oxidative stress is a promising strategy for alleviating ALI. 1,2,3,4,6-<i>O</i>-pentagalloylglucose (PGG) has a pronounced therapeutic effect on ALI, with excellent anti-inflammatory and antioxidant effects. However, poor solubility and low bioavailability have affected its clinical application. In this study, carrier-free PGG nanoparticles (PGG NPs) were prepared by antisolvent precipitation method. PGG NPs have been engineered to improve solubility, sustained release behavior, and higher bioavailability than free PGG. Moreover, the pharmacodynamic results showed that the remarkable protective effect of PGG NPs on ALI in rats is better than that of free PGG, which is related to the activation of Nrf2/Keap1/HO-1/NLRP3 pathway. Overall, this study not only demonstrates the efficacy and safety of PGG against ALI, but also holds promise as a carrier-free nanodrug system.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 5","pages":"1121–1131 1121–1131"},"PeriodicalIF":4.0,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144097951","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Super-Sensitive Chemiluminescent Probe for the Detection of Caspase-3 Activity","authors":"Rozan Tannous,&nbsp;Chi Zhang and Doron Shabat*,&nbsp;","doi":"10.1021/acs.bioconjchem.5c0015110.1021/acs.bioconjchem.5c00151","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.5c00151https://doi.org/10.1021/acs.bioconjchem.5c00151","url":null,"abstract":"<p >Caspase-3 is a pivotal enzyme in the apoptosis pathway that is responsible for executing programmed cell death through the cleavage of key cellular proteins. Existing fluorescence-based probes for caspase-3 detection suffer from limitations such as background noise from tissue autofluorescence and light scattering, reducing their sensitivity and real-time imaging capabilities. To overcome these limitations, we developed a chemiluminescent probe, Ac-DEVD-CL, that enables the highly sensitive and selective detection of caspase-3 activity. Upon caspase-3-mediated cleavage, the probe undergoes a self-immolative reaction that triggers a chemiluminescent signal, allowing real-time monitoring of the enzymatic activity. Probe Ac-DEVD-CL demonstrated an exceptionally high turn-on response, with a 5000-fold increase in the chemiluminescent signal upon enzymatic activation. The probe exhibited notable specificity for caspase-3, with minimal cross-reactivity toward other biologically relevant proteases and tumor-associated enzymes. Additionally, inhibition studies using the caspase-3 inhibitor confirmed that the probe’s activation is exclusively mediated by caspase-3. A direct comparison with the commercially available fluorescent probe revealed that probe Ac-DEVD-CL offers significantly improved sensitivity, achieving a signal-to-noise ratio 380-fold higher and a limit of detection 100-fold lower. These results establish probe Ac-DEVD-CL as a highly effective tool for detecting caspase-3 activity with superior precision. Finally, we validated the probe’s utility in imaging apoptosis in live cells. In 4T1 breast cancer cells treated with cisplatin, Ac-DEVD-CL generated a strong chemiluminescent signal, with a three-order-of-magnitude enhancement compared to untreated cells. Overall, the probe Ac-DEVD-CL demonstrates a significant improvement in detection sensitivity, providing a powerful and versatile chemiluminescent probe for real-time imaging of caspase-3 activity. Its exceptional sensitivity and selectivity could make it a valuable tool for cancer research, drug discovery, and therapeutic monitoring.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 5","pages":"1113–1120 1113–1120"},"PeriodicalIF":4.0,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acs.bioconjchem.5c00151","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144098193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}