Bioconjugate Chemistry最新文献

{"title":"Delivery of Tempol from Polyurethane Nanocapsules to Address Oxidative Stress Post-Injury","authors":"Temitope Ale,&nbsp;Tolulope Ale,&nbsp;Kimberly J. Baker,&nbsp;Kameel M. Zuniga,&nbsp;Jack Hutcheson and Erin Lavik*,&nbsp;","doi":"10.1021/acs.bioconjchem.4c0036010.1021/acs.bioconjchem.4c00360","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.4c00360https://doi.org/10.1021/acs.bioconjchem.4c00360","url":null,"abstract":"<p >Traumatic brain injuries (TBIs) result in significant morbidity and mortality due to the cascade of secondary injuries involving oxidative stress and neuroinflammation. The development of effective therapeutic strategies to mitigate these effects is critical. This study explores the fabrication and characterization of polyurethane nanocapsules for the sustained delivery of Tempol, a potent antioxidant. The nanocapsules were designed to extend the release of Tempol over a 30-day period, addressing the prolonged oxidative stress observed post-TBI. Tempol-loaded polyurethane nanocapsules were synthesized using interfacial polymerization and nanoemulsion techniques. Two generations of nanocapsules were produced, differing in Tempol loading and PEGylation levels. The first generation, with lower Tempol loading, exhibited an average size of 159.8 ± 12.61 nm and a Z-average diameter of 771.9 ± 87.95 nm. The second generation, with higher Tempol loading, showed an average size of 141.4 ± 6.13 nm and a Z-average diameter of 560.7 ± 171.1 nm. The zeta potentials were −18.9 ± 5.02 mV and −11.9 ± 3.54 mV for the first and second generations, respectively. Both generations demonstrated the presence of urethane linkages, confirmed by Fourier Transform Infrared Spectroscopy (FTIR). Loading studies revealed Tempol concentrations of 61.94 ± 3.04 μg/mg for the first generation and 77.61 ± 3.04 μg/mg for the second generation nanocapsules. Release profiles indicated an initial burst followed by a sustained, nearly linear release over 30 days. The higher PEGylation in the second generation nanocapsules is advantageous for intravenous administration, potentially enhancing their therapeutic efficacy in TBI treatment. This study demonstrates the feasibility of using polyurethane nanocapsules for the prolonged delivery of Tempol, offering a promising approach to manage oxidative stress and improve outcomes in TBI patients. Future work will include testing these nanocapsules in vivo to determine their potential at modulating recovery from TBI.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 2","pages":"146–151 146–151"},"PeriodicalIF":4.0,"publicationDate":"2025-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143435936","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":"Method for Screening Sodium Cyanoborohydride for Free Cyanide Content and Its Impact on Bioconjugation Chemistry","authors":"Jarrod P. Cohen*,&nbsp;Adam DiCaprio,&nbsp;Jian He,&nbsp;Mikhail Reibarkh,&nbsp;James Small and Matthew Schombs,&nbsp;","doi":"10.1021/acs.bioconjchem.4c0051410.1021/acs.bioconjchem.4c00514","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.4c00514https://doi.org/10.1021/acs.bioconjchem.4c00514","url":null,"abstract":"<p >Sodium cyanoborohydride (CBH) is commonly used as a mild reducing agent in the reductive amination of aldehydes and free amines. Within the pharmaceutical industry, this reaction is employed in the bioconjugation of proteins and peptides. Free cyanide species such as HCN and NaCN are known residual impurities in CBH that can contribute to the formation of undesired side products including cyanoamines and cyanohydrins. In commercial processes, the potential for bound cyanated species requires an analytical control strategy to monitor and mitigate any risk to human health. Given these concerns, minimization of cyanated side products is of utmost priority and can be achieved through a robust control strategy of quantitative screening of starting materials for free cyanide. Alternative risk mitigation strategies such as purification of bound cyanide containing species to pure species are less effective due to minor chemical differences between the expected product and bound cyanide species. Herein, we present a simple chromatographic assay for the quantitation of free cyanide in the raw material sodium cyanoborohydride. Method development, robustness evaluation, and scientific soundness assessment are reported with excellent linearity, accuracy, precision, and specificity. Additionally, this method was applied for the evaluation of raw material supplied from 10 commercial sources, none of which report a specification for free cyanide within their certificate of analysis. The measured free cyanide from these vendors ranged from 8 to 80 mM concentration, thereby confirming the value of screening these raw materials. Finally, we demonstrate the impact of free cyanide on a model bioconjugation reaction between ornithine and glyceraldehyde.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 2","pages":"245–252 245–252"},"PeriodicalIF":4.0,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acs.bioconjchem.4c00514","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143436010","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}

{"title":"Development of Transiently Strainable Benzocycloheptenes for Catalyst-Free, Visible-Light-Mediated [3 + 2]-Cycloadditions","authors":"Shivangi Kharbanda,&nbsp;Osaid Alkhamayseh,&nbsp;Georgia Eastham and Jimmie D. Weaver*,&nbsp;","doi":"10.1021/acs.bioconjchem.4c0059510.1021/acs.bioconjchem.4c00595","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.4c00595https://doi.org/10.1021/acs.bioconjchem.4c00595","url":null,"abstract":"<p >Dynamic photogeneration of ephemeral and reactive species is enabling for chemical reactions, providing spatial and temporal control. A previous study from our group established the ability of 6,7-dihydro-5H-benzo[7]annulene, benzocycloheptene (<b>BC7</b>), to convert photochemical energy into ring strain, enabling the rapid cycloaddition of alkyl azides with the reversibly formed and transient <i>trans</i>-isomer, affording versatile nonaromatic triazolines. Despite the conceptual advances of the previous study, some challenges remained: the fragility of the triazoline products, the low regioselectivity for the cycloaddition, a need for an iridium-based photosensitizer and organic-based solvents, and a lack of convenient linchpin functional group handles. Herein, we communicate the development of a second generation of <b>BC7</b> molecules that overcome the issues of the first generation. A method to convert fragile triazoline products to stable triazoles was developed. The alkene component was polarized with a carbonyl group, dramatically improving the regioselectivity while simultaneously red-shifting the absorbance of the cycloalkene into the visible region, which was expected to facilitate direct excitation and eliminate the need for photocatalysts. However, experiments indicated that the cycloaddition involved passage through a triplet manifold, complicating the direct excitation strategy. This was successfully overcome by attaching a bromine atom directly to the alkene moiety, which accelerated singlet-to-triplet intersystem crossing by the heavy atom effect. Further exploration identified sites of substitution that can increase the water solubility and provide a handle for the loading of chemical tools and probes.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 2","pages":"302–308 302–308"},"PeriodicalIF":4.0,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acs.bioconjchem.4c00595","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143436341","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}

{"title":"A Unique Prodrug Targeting the Prostate-Specific Membrane Antigen for the Delivery of Monomethyl Auristatin E","authors":"Hunter N. Bomba,&nbsp;Melody D. Fulton,&nbsp;Emily A. Savoy,&nbsp;Beatrice Langton-Webster* and Clifford E. Berkman,&nbsp;","doi":"10.1021/acs.bioconjchem.4c0029710.1021/acs.bioconjchem.4c00297","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.4c00297https://doi.org/10.1021/acs.bioconjchem.4c00297","url":null,"abstract":"<p >Monomethyl auristatin E (MMAE) is a promising treatment option for patients diagnosed with prostate cancer (PCa); however, toxicities prevent MMAE from being administered as free drug. No MMAE-based treatment is currently marketed for PCa. Herein, we describe a small-molecule-drug conjugate, CTT2274, for the selective delivery of MMAE. CTT2274 is composed of a prostate-specific membrane antigen (PSMA)-binding scaffold, a biphenyl motif, a pH-sensitive phosphoramidate linker, and MMAE payload. We demonstrate that CTT2274 shows selective binding to PSMA, which is overexpressed on PCa cells, and induces tumor cell death <i>in vitro</i>. In a patient-derived xenograft tumor model of PCa in mice, we show that weekly intravenous dosing of CTT2274 at 3.6 mg/kg for six weeks is superior to treatment with free MMAE at equivalent doses. Mice treated with CTT2274 experienced prolonged tumor suppression and significantly greater overall survival than mice treated with PBS. Additionally, the safety of CTT2274 compared to an equivalent dose of MMAE was assessed in healthy, non-tumor-bearing mice. Our results demonstrate that CTT2274 therapy is as efficacious as MMAE, results in superior overall survival, and has a more favorable safety profile. Together, these data indicate that CTT2274 is a candidate for clinical translation for the treatment of PCa.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 2","pages":"169–178 169–178"},"PeriodicalIF":4.0,"publicationDate":"2025-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143436035","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":"Nanoscale Effects in the Room-Temperature UV–Visible Photoluminescence from Silica Particles and Its Cancer Cell Imaging","authors":"Divya Rani,&nbsp;Deepika Singh,&nbsp;Anil Kumar,&nbsp;Monika Dhiman,&nbsp;Anjali Saini,&nbsp;Partho Biswas,&nbsp;Rachana Rachana,&nbsp;Partha Roy,&nbsp;Mrinal Dutta and Arup Samanta*,&nbsp;","doi":"10.1021/acs.bioconjchem.4c0042010.1021/acs.bioconjchem.4c00420","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.4c00420https://doi.org/10.1021/acs.bioconjchem.4c00420","url":null,"abstract":"<p >Silica nano/microparticles have generated significant interest for the past decades, emerging as a versatile material with a wide range of applications in photonic crystals, bioimaging, chemical sensors, and catalysis. This study focused on synthesizing silica nano/microparticles ranging from 20 nm to 1.2 μm using the Stöber and modified Stöber methods. The particles exhibited photoluminescence emission across a UV–visible range, specifically in the UV (∼290, ∼327, ∼339, and ∼377 nm), blue (∼450 nm), green (∼500 nm), yellow (∼576 nm), and red (∼634 nm) range of the electromagnetic spectrum. These emissions are due to radiative relaxation processes involving oxygen-deficient centers arising due to unrelaxed oxygen vacancies, strong interacting surface silanols, 2-fold coordinated silicon, self-trapped excitons, hydrogen-related species, strain-induced defects, and nonbridging oxygen hole centers excited via two-photon and single photon absorption. The increased PL intensity with a decreasing particle size was attributed to higher concentrations of defect sites in the case of smaller-sized particles. The MTT assay, AO/EB staining, and the DCFDA assay confirmed the biocompatible nature of silica particles in the HepG2 cell line. In addition, the cell viability assay in a normal cell line (HEK293) also showed no substantial cell death. Successful bioimaging of HepG2 cells was performed with silica nano/microparticles, which exhibited blue and green fluorescence, along with Hoechst33258 dye. Even though 20 nm-sized silica particles showed higher PL emission, particles sized above 20 nm showed better fluorescence in HepG2 cells, citing their potential in <i>in vitro</i> bioimaging applications.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 2","pages":"203–215 203–215"},"PeriodicalIF":4.0,"publicationDate":"2025-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143436046","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":"Biomechanical and Functional Features of the Carrier Erythrocytes Prolonging Circulation Time of Biotherapeutic Targeted to Glycophorin A","authors":"Alina D. Peshkova,&nbsp;Taylor V. Brysgel,&nbsp;Parth Mody,&nbsp;Jia Nong,&nbsp;Zhicheng Wang,&nbsp;Jacob W. Myerson,&nbsp;Rustem I. Litvinov,&nbsp;John W. Weisel,&nbsp;Jacob S. Brenner,&nbsp;Patrick M. Glassman*,&nbsp;Oscar A. Marcos-Contreras* and Vladimir R. Muzykantov*,&nbsp;","doi":"10.1021/acs.bioconjchem.4c0052210.1021/acs.bioconjchem.4c00522","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.4c00522https://doi.org/10.1021/acs.bioconjchem.4c00522","url":null,"abstract":"<p >Red blood cells (RBCs) serve as natural transporters and can be modified to enhance the pharmacokinetics and pharmacodynamics of a protein cargo. Affinity targeting of Factor IX (FIX) to the RBC membrane is a promising approach to improve the (pro)enzyme’s pharmacokinetics. For RBC targeting, purified human FIX was conjugated to the anti-mouse glycophorin A monoclonal antibody Ter119. The goal of this study was to characterize the activity of the FIX-Ter119 conjugate and efficacy of its loading on RBCs, as well as to investigate the biodistribution, pharmacokinetics, and various biological properties of the loaded RBCs. Mouse RBCs were incubated with the Ter119-FIX conjugate, where adding 10,000 molecules per RBC resulted in 37% binding (4K/RBC), and 50,000 molecules per RBC resulted in 34% binding (17K/RBC). The pharmacokinetics (PK) profile showed that more than 90% of the Ter119-FIX conjugate was associated with RBCs and circulated stably bound to the RBCs for 24 h, increasing the area under the PK curve 7.6 times vs free FIX. Ter119-FIX loaded RBCs have specific procoagulant FIXa activity, including promotion of thrombin generation and acceleration of clotting in FIX-deficient plasma. Morphological characterization shows that Ter119-FIX-loaded RBCs undergo a shape change, with an increased fraction of echinocytes and spheroidal RBCs. Ektacytometry and electron microscopy assessment of RBC compressibility reveal a dose-dependent reduction in the deformability of RBCs loaded with Ter119-FIX. In conclusion, RBCs loaded with Ter119-FIX have the potential to serve as prohemostatic agents, but their reduced deformability warrants further engineering of Ter119-FIX to improve the safety profile.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 2","pages":"263–275 263–275"},"PeriodicalIF":4.0,"publicationDate":"2025-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143435946","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":"Synthesis and Biological Evaluation of Bile Acid–Triclosan Conjugates: A Study on Antibacterial, Antibiofilm, and Molecular Docking","authors":"Neha V. Rathod,&nbsp; and ,&nbsp;Satyendra Mishra*,&nbsp;","doi":"10.1021/acs.bioconjchem.4c0053910.1021/acs.bioconjchem.4c00539","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.4c00539https://doi.org/10.1021/acs.bioconjchem.4c00539","url":null,"abstract":"<p >This work describes the synthesis, characterization, and antibacterial properties of four bile acid–triclosan conjugates. The in vitro antibacterial activity of synthetic bile acid–triclosan conjugates was investigated against a panel of Gram-positive and Gram-negative bacteria. Conjugates <b>3</b> and <b>4</b> show high activity against <i>Escherichia coli</i> (ATCC25922), with IC₅₀ values of 2.94 ± 0.7 and 1.51 ± 0.05 μM, respectively. Conjugate <b>4</b> demonstrated 9 times the activity of triclosan (6.77 μM) and 18 times the potency of kanamycin, a well-known antibiotic. Compound <b>3</b> showed higher potential activity against all evaluated strains, including <i>Bacillus megaterium</i> (IC₅₀: 3.05 ± 0.02), <i>Bacillus amyloquefaciens</i> (IC₅₀: 8.79 ± 0.01), <i>Serratia marcescens</i> (IC₅₀: 6.77 ± 0.4), and <i>E. coli</i> (IC₅₀: 1.51 ± 0.05 μM). These findings indicate that it has broad-spectrum antibacterial activity. Bile acid–triclosan conjugates prevent biofilms by up to 99% at low doses (conjugates <b>4</b>; 4.16 ± 0.8 μM), compared to triclosan. Conjugate <b>5</b> was most potent against <i>B. amyloquefaciens</i> (IC₅₀ = 5.23 ± 0.2 μM), while conjugate <b>4</b> was most effective against <i>B. megaterium</i> (IC₅₀ = 4.16 ± 0.8 μM) in biofilm formation. These conjugates inhibit biofilm formation by limiting the extracellular polymeric substance generation. The in vitro antibacterial study revealed that bile acid–triclosan conjugates were more effective than the parent molecule triclosan at inhibiting bacterial growth and biofilm formation against both Gram-positive and Gram-negative bacteria.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 2","pages":"276–290 276–290"},"PeriodicalIF":4.0,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143436282","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":"Effect of Lipidation on the Structure, Oligomerization, and Aggregation of Glucagon-like Peptide 1","authors":"Eva Přáda Brichtová,&nbsp;Irina A. Edu,&nbsp;Xinyang Li,&nbsp;Frederik Becher,&nbsp;Ana L. Gomes dos Santos and Sophie E. Jackson*,&nbsp;","doi":"10.1021/acs.bioconjchem.4c0048410.1021/acs.bioconjchem.4c00484","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.4c00484https://doi.org/10.1021/acs.bioconjchem.4c00484","url":null,"abstract":"<p >Lipidated analogues of glucagon-like peptide 1 (GLP-1) have gained enormous attention as long-acting peptide therapeutics for type 2 diabetes and also antiobesity treatment. Commercially available therapeutic lipidated GLP-1 analogues, semaglutide and liraglutide, have the great advantage of prolonged half-lives <i>in vivo</i> of hours and days instead of minutes as is the case for native GLP-1. A crucial factor in the development of novel lipidated therapeutic peptides is their physical stability, which greatly influences manufacturing and drug product development. This work provides a systematic study of the solubility, structure, oligomerization, and long-term stability of five different lipidated analogues of GLP-1, varying in the position of the lipidation site and the nature of lipid attachment. The lipidation was found to negatively impact the peptide solubility, in all cases, limiting it to a specific pH range. An increase in the α-helical secondary structure was observed upon lipidation, and the lipidated analogues were found to form larger and more stable oligomeric species compared to nonlipidated GLP-1. Importantly, the distributions and populations of oligomeric species formed were regulated by both the position and the nature of the lipidation. During the 6 days of sample aging, several lipidated analogues formed aggregates with variable morphologies ranging from elongated mature fibrils to amorphous structures. The kinetics of aggregation often showed multiple steps and did not follow a standard nucleation-propagation mechanism. A wide range of behaviors was observed, and while our observations indicate that the formation of a single stable oligomer results in the greatest physical stability, positioning the lipid group toward the N-terminus of the peptide results in extremely rapid amyloid formation. We believe that our study provides important findings for the development of long-acting lipidated analogues of peptide therapeutics.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 3","pages":"401–414 401–414"},"PeriodicalIF":4.0,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acs.bioconjchem.4c00484","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143641373","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}

{"title":"Modular Synthesis of Anti-HER2 Dual-Drug Antibody-Drug Conjugates Demonstrating Improved Toxicity","authors":"Christine S. Nervig,&nbsp;Megan Rice,&nbsp;Marcello Marelli,&nbsp;R. James Christie and Shawn C. Owen*,&nbsp;","doi":"10.1021/acs.bioconjchem.4c0039810.1021/acs.bioconjchem.4c00398","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.4c00398https://doi.org/10.1021/acs.bioconjchem.4c00398","url":null,"abstract":"<p >Antibodies have gained clinical success in the last two decades for the targeted delivery of highly toxic small molecule chemotherapeutics. Yet antibody-drug conjugates (ADCs) often fail in the clinic due to the development of resistance. The delivery of two mechanistically distinct small molecule drugs on one antibody is of increasing interest to overcome these challenges with single-drug ADCs. We have developed a modular synthetic strategy for the construction of a library of 19 dual-drug ADCs where drugs are conjugated through unnatural cyclopentadiene-containing amino acids and native cysteine residues on an anti-HER2 trastuzumab scaffold. Importantly, this strategy utilizes the same functional group on the linker-drug construct; this allows for the facile addition of drugs at either conjugation site and enables the evaluation of different drug-to-antibody ratios and combinations of drug pairs. We tested the library on high- and mid-HER2 expressing cell lines and observed increased toxicity in several dual-drug ADCs compared with single-drug constructs. The strategy developed herein provides a method for the facile synthesis, characterization, and evaluation of dual-payload ADCs. Simultaneous delivery of combinations of drugs with distinct mechanisms of action is critical for the next generation of targeted drug delivery.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 2","pages":"190–202 190–202"},"PeriodicalIF":4.0,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143436284","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":"Quantum Dot Erythropoietin Bioconjugates Enhance EPO-Receptor Clustering on Transfected Human Embryonic Kidney Cells","authors":"Ryan N. Porell,&nbsp;Okhil K. Nag,&nbsp;Michael H. Stewart,&nbsp;Kimihiro Susumu,&nbsp;Eunkeu Oh and James B. Delehanty*,&nbsp;","doi":"10.1021/acs.bioconjchem.4c0052110.1021/acs.bioconjchem.4c00521","DOIUrl":"https://doi.org/10.1021/acs.bioconjchem.4c00521https://doi.org/10.1021/acs.bioconjchem.4c00521","url":null,"abstract":"<p >Erythropoietin (EPO)-induced cellular signaling through the EPO receptor (EPOR) is a fundamental pathway for the modulation of cellular behavior and activity. In our previous work, we showed in primary human astrocytes that the multivalent display of EPO on the surface of semiconductor quantum dots (QDs) mediates augmented JAK/STAT signaling, a concomitant 1.8-fold increase in the expression of aquaporin-4 (AQPN-4) channel proteins, and a 2-fold increase in the AQPN-4-mediated water transport activity. Our hypothesis is that this enhanced signaling involves the simultaneous ligation and clustering of EPOR by QD-EPO conjugates. Here, we utilized a human embryonic kidney (HEK 293T/17) cell line transfected with EPOR fused to enhanced green fluorescent protein (eGFP) to visualize EPOR clustering. We demonstrate that QDs displaying five copies of EPO (bearing a C-terminal 6-histidine tract) on the nanoparticle surface induce a 1.8-fold increase in EPOR clustering compared to monomeric EPO at the same concentration. Our findings confirm the critical role played by the multivalent display of EPO in mediating clustering of the EPOR. More generally, these results illustrate the capability of nanoparticle-growth factor bioconjugates to control the activity of cognate receptors and the important role played by multivalent display in the modulation of selective cellular delivery and signaling.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":"36 2","pages":"160–168 160–168"},"PeriodicalIF":4.0,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143436293","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}