Archives of biochemistry and biophysics最新文献

{"title":"A fluorescent cholesterol analog, R-Chol, mimics the dynamics of free cholesterol in live cells","authors":"Hinako Mogi,&nbsp;Yumi Yamashita,&nbsp;Masumi Sano,&nbsp;Ryo Kadowaki,&nbsp;Takuya Honda,&nbsp;Takafumi Kohama,&nbsp;Hiroyuki Nakamura","doi":"10.1016/j.abb.2025.110600","DOIUrl":"10.1016/j.abb.2025.110600","url":null,"abstract":"<div><div>Cholesterol is an essential component of the mammalian cell membrane. Elucidating the dynamics of intracellular cholesterol is extremely important for understanding the mechanisms underlying life phenomena and diseases. The fluorescent cholesterol analog, R-Chol, has a fluorescent group at the 3-OH position of free cholesterol, which is less toxic and helpful for studying free cholesterol trafficking in live cells. In the present study, we determined the details of R-Chol dynamics in live cells. R-Chol was taken up by the plasma membrane within 10 min and then incorporated into the cells by endocytosis. Free cholesterol in late endosomes/lysosomes is transported to the endoplasmic reticulum via NPC1 proteins. In <em>Npc1</em>-deficient CHO cells, R-Chol accumulates in late endosomes/lysosomes. R-Chol accumulation in <em>Npc1</em>-deficient cells was attenuated by the overexpression of Rab9, which regulates intracellular transport from late endosomes to the Golgi apparatus. These results indicate that R-Chol incorporated via endocytosis is transported to late endosomes/lysosomes and then to the Golgi apparatus and endoplasmic reticulum by NPC1-dependent and NPC1-independent vesicular trafficking.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110600"},"PeriodicalIF":3.0,"publicationDate":"2025-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144904366","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bile acids and polyphenols inhibit succinic anhydride-induced protein succinylation and amyloid aggregation: Mechanistic insights","authors":"Yilin Huang,&nbsp;Xingli Huo,&nbsp;Huijun Liu,&nbsp;Danni Li,&nbsp;Zongning Yin","doi":"10.1016/j.abb.2025.110601","DOIUrl":"10.1016/j.abb.2025.110601","url":null,"abstract":"<div><div>Lysine succinylation is a major post-translational modification affecting diverse proteins, and its excessive occurrence can lead to protein misfolding and aggregation—hallmarks of various proteinopathies, such as amyloid-β and Tau tangle formation in Alzheimer's disease and islet amyloid polypeptide aggregation in type 2 diabetes. Here, we investigated the inhibitory effects of bile acid metabolites (deoxycholic acid, glycocholic acid, and taurocholic acid sodium) and natural polyphenols (anthocyanin and salidroside) on succinylation and succinylation-induced amyloid aggregation. Succinylation levels were evaluated using the ninhydrin assay before and after treatment, and aggregation behavior and structural alterations were characterized by SDS-PAGE, inverted fluorescence microscopy, and intrinsic fluorescence spectroscopy. Additionally, fluorescence quenching and molecular docking were used to explore the underlying mechanisms. All five small molecules significantly reduced succinylation in a concentration-dependent manner (p &lt; 0.05). Among bile acids, taurocholic acid sodium exhibited the strongest suppression of aggregation (40.99 %), followed by glycocholic acid (28.32 %) and deoxycholic acid (27.94 %). Anthocyanin showed greater inhibition (33.81 %) than salidroside (26.54 %) (all p &lt; 0.05). The results suggest that these small molecules inhibit succinylation-induced aggregation potentially by interacting with proteins and altering their conformations, thereby preventing excessive succinylation at lysine residues. This study provides new insight into the interplay between bile acid metabolism and protein homeostasis and highlights the therapeutic potential of natural compounds in preventing protein aggregation-related diseases.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110601"},"PeriodicalIF":3.0,"publicationDate":"2025-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144904224","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Unraveling the mechanisms of spinigerin: A promising antimicrobial peptide against antibiotic resistance through molecular dynamics simulations","authors":"Masoud Shahriari ,&nbsp;Leila Fathollahzadeh ,&nbsp;Yasaman Mahmoodi ,&nbsp;Nasim Ahmadian ,&nbsp;Farahnoosh Doustdar ,&nbsp;Reza Talandashti ,&nbsp;Faramarz Mehrnejad","doi":"10.1016/j.abb.2025.110602","DOIUrl":"10.1016/j.abb.2025.110602","url":null,"abstract":"<div><div>The rise of infectious diseases driven by antibiotic resistance has become a critical global concern, prompting the search for alternative therapeutic strategies. Among these, antimicrobial peptides (AMPs) have emerged as promising candidates. Spinigerin, a potent AMP, has demonstrated significant antibacterial and antifungal activity. Its mechanism involves disrupting microbial cell membranes by forming transmembrane pores that induce cell lysis. However, the precise mechanisms underlying pore formation remain poorly understood. To explore spinigerin's pore-forming potential, a comprehensive study was conducted using five different membrane models. A combination of coarse-grained (CG) and all-atom (AA) molecular dynamics simulations, along with umbrella sampling, was employed to elucidate the interactions between spinigerin and the membrane models. The results revealed that the peptide induces notable structural changes in the anionic and hybrid membranes, such as increased curvature, lipid reorganization, and altered membrane thickness. In contrast, spinigerin maintains stability in zwitterionic membranes with minimal disruption. Additionally, pore dimension analysis indicated that the initial stage of pore formation was more feasible and stable in the anionic and zwitterionic-anionic membrane models compared to the zwitterionic membrane. These findings underscore the critical role of lipid composition in determining the efficacy and mechanism of action of AMPs like spinigerin. Outstandingly, spinigerin's selective ability to target bacterial membranes while sparing host cells highlights its strong potential as a therapeutic agent.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110602"},"PeriodicalIF":3.0,"publicationDate":"2025-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144907281","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhibition of ferroptosis by serine protease inhibitor attenuates acute respiratory distress syndrome","authors":"Qinyue Guo ,&nbsp;Xiaoming Gao ,&nbsp;Jiajia Ren ,&nbsp;Guorong Deng ,&nbsp;Ruohan Li ,&nbsp;Chuchu Zhang ,&nbsp;Xi Xu ,&nbsp;Jiamei Li ,&nbsp;Wanyuan Liu ,&nbsp;Lingzhi Qin ,&nbsp;Gang Wang","doi":"10.1016/j.abb.2025.110596","DOIUrl":"10.1016/j.abb.2025.110596","url":null,"abstract":"<div><h3>Background</h3><div>Acute respiratory distress syndrome (ARDS), characterized by high mortality, involves multiple molecular programs, notably ferroptosis—a form of immunogenic cell death driven by iron overload and lipid peroxidation. Ulinastatin (UTI), a serine protease inhibitor, shows clinical efficacy in ARDS, but its underlying mechanism remains unclear. We aim to identify potential molecular targets in this process to promote clinical translation in ARDS treatment.</div></div><div><h3>Methods</h3><div>We performed RNA sequencing (RNA-seq) on lung tissues from LPS-induced ARDS mice revealed significant enrichment of ferroptosis-related pathways in UTI-treated ARDS mice, prompting the hypothesis that UTI mitigates ARDS by suppressing ferroptosis. Using LPS-induced murine ARDS, we assessed UTI's therapeutic effects via histopathology, qRT-PCR, RNA sequencing, and molecular assays. Ferroptosis biomarkers (iron, MDA, GSH), key proteins (GPX4, KEAP1, NRF2), and inflammatory cytokines were evaluated. In vitro, HUVEC and MLE-12 were used to investigate the molecular mechanisms by which UTI's functions via ferroptosis. Molecular docking explored UTI-KEAP1/NRF2 interactions.</div></div><div><h3>Results</h3><div>UTI significantly attenuated lung injury, reduced inflammatory cytokines (IL-1β, IL-6, TNF-α), and restored hepatic/renal function in LPS-challenged mice. Transcriptomics revealed ferroptosis as a top enriched pathway suppressed by UTI. Mechanistically, in both HUVEC and MLE-12 cells, UTI attenuated LPS-induced increases in labile iron, MDA, and lipid ROS levels. Additionally, UTI suppressed KEAP1 expression while activating NRF2, an effect comparable to that of ferroptosis inhibitors. Consequently, GPX4 expression was upregulated, suggesting a potential anti-ferroptotic mechanism.</div></div><div><h3>Conclusion</h3><div>Inhibition of ferroptosis is a novel mechanism underpinning UTI's lung-protective effect against ARDS. UTI potentially regulates the KEAP1-NRF2 interaction through direct binding to KEAP1, offering a new molecular-level explanation for its mechanism of action.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110596"},"PeriodicalIF":3.0,"publicationDate":"2025-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144904367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis, DFT and molecular docking studies of N-Heterocyclic carbene selenium compounds conferring anticancer and antibacterial activity","authors":"Ahmad Hassan ,&nbsp;Rizwan Ashraf ,&nbsp;Muhammad Adnan Iqbal ,&nbsp;Khizar Hayat ,&nbsp;Faisal Jamil ,&nbsp;Saed Ayidh Althobaiti ,&nbsp;Syeda Anjum Tahira ,&nbsp;Mohamed El-Naggar ,&nbsp;Mohamed Mohamed Soliman","doi":"10.1016/j.abb.2025.110598","DOIUrl":"10.1016/j.abb.2025.110598","url":null,"abstract":"<div><div>Target based drug design is an important strategy that increases the selectivity, efficacy and safety of drug candidates. In this study we designed synthesis of benz-imidazolium salts (L1-L3) and selenium compounds (C1–C3) to investigate their enzyme inhibition potential and bioactivity. Successful synthesis was confirmed through analytical techniques like UV–Vis., FTIR, ¹H &amp; ¹³C NMR and mass spectrometry that further supported by computational (DFT) studies. Molecular docking studies of C1–C3 against key molecular targets (COX-1, EGF, VEGF and HIF) was conducted. Among the test compounds C1 showed an impressive binding affinity of −6.14 kcal mol⁻¹ against EGF which is comparable to standard drug 5-FU (−4.97 kcal mol⁻¹). The validation of docking results through <em>in vitro</em> studies confirmed C1 as most potent inhibitor among the test compounds, having inhibition of 67.4 ± 1.3 % and 86.7 ± 1.8 % against COX-1 and EGF respectively. Furthermore, test compounds showed significant inhibition potential against thioredoxin reductase (TrxR). Cytotoxicity profiling across HepG2, HeLa and A-2780 cell lines confirmed C1 as the lead compound, with IC₅₀ values of 0.956, 1.986, and 0.862 μg/mL, respectively. Test compounds also showed antibacterial activity by showing inhibition zone 8.5 ± 1.1–27.0 ± 1.2 mm against <em>E. coli</em> and <em>S. aureus</em>. These findings showed that NHC based selenium compounds could be a potential drug candidate for chemotherapy against multiple cancerous strains.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110598"},"PeriodicalIF":3.0,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144895776","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring the mechanical and electrical properties of cancer cells for improved diagnosis and treatment","authors":"Michał Gebuza ,&nbsp;Julia Górczyńska ,&nbsp;Wiktoria Paczos ,&nbsp;Szymon Pieczara ,&nbsp;Katarzyna Rekiel ,&nbsp;Piotr Urbanowicz ,&nbsp;Marek Kulbacki ,&nbsp;Vitalij Novickij ,&nbsp;Julita Kulbacka","doi":"10.1016/j.abb.2025.110599","DOIUrl":"10.1016/j.abb.2025.110599","url":null,"abstract":"<div><div>Over the past decade, there has been growing interest in the mechanical and electrical properties of cancer cells and their potential for improved diagnosis and treatment. Cancer cells exhibit unique mechanical and electrical properties, including altered stiffness, adhesion, charge, and shape, which distinguish them from normal cells. Researchers are exploring ways to use these properties to develop new diagnostic tools and therapeutic approaches that selectively target cancer cells while minimizing harm to healthy tissue. Recent advancements in machine learning (ML) have enhanced the analysis of cancer cell behavior, enabling more accurate identification based on electrical and mechanical “biomarkers” and offering the potential for early diagnosis and personalized treatment strategies. In this review, we will examine the most recent research on the mechanical and electrical properties of cancer cells and discuss their potential implications for enhancing the diagnosis and treatment of cancer.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110599"},"PeriodicalIF":3.0,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144889211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inula britannica exerts antioxidant and anti-inflammatory effects in SH-SY5Y cells through the Nrf2–Keap1 signaling pathway","authors":"Hyun-Joo Yoon,&nbsp;Ji-Young Park,&nbsp;Su-Jin Min,&nbsp;Eun-Soo Lee,&nbsp;Na-Kyoung Lee,&nbsp;Hyun-Dong Paik","doi":"10.1016/j.abb.2025.110597","DOIUrl":"10.1016/j.abb.2025.110597","url":null,"abstract":"<div><div>The yellow flower of the <em>Asteraceae</em> family, <em>Inula britannica</em> (<em>IB</em>) of have been used as a medicinal herb since ancient times. Modern studies have widely known the anti-inflammatory effects and antioxidant effects of <em>IB</em>. Therefore, this study aimed to analyze the neuroprotective effects of <em>IB</em> extract based on its antioxidant and anti-inflammatory effects and to assess its potential for development as a functional material. After preparing the extract of <em>IB</em> based on the solvent with water and 70 % ethyl alcohol (EtOH), the phytochemical components were identified. The effects of the <em>IB</em> extracts on the Nrf2–Keap1 signaling pathway, NF-κB signaling pathway, and Bax/Bcl-2 ratio were investigated in H₂O₂-induced SH-SY5Y cells using reverse transcription-polymerase chain reaction and western blotting. Both <em>IB</em> extracts activated the representative antioxidant signaling pathway Nrf2–Keap1 in a concentration-dependent manner. In the inflammation-related NF-κB signaling pathway, the expression of each factor was inhibited, and the Bax/Bcl-2 ratio, which is related to apoptosis, decreased upon <em>IB</em> extract treatment. Phytochemical analysis using liquid chromatography-tandem mass spectrometry identified polyphenols and flavonoids as the primary active compounds in <em>IB</em>. These findings indicate that <em>IB</em> extract has potential as a functional material for the treatment of neurodegenerative diseases.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110597"},"PeriodicalIF":3.0,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144889405","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Discovery of a novel CYP107E41 from Streptoalloteichus sp. KCCM40925 for advancing steroid hydroxylation catalysis","authors":"Prakash Paudel ,&nbsp;Kamal Prasad Regmi ,&nbsp;Ki-Hwa Kim ,&nbsp;Tae-Jin Oh","doi":"10.1016/j.abb.2025.110595","DOIUrl":"10.1016/j.abb.2025.110595","url":null,"abstract":"<div><div>Cytochrome P450 enzymes (CYPs) are versatile biocatalysts involved in the biosynthesis, activation, and detoxification of a wide range of endogenous and exogenous compounds. While the CYP107 family is primarily associated with macrolide antibiotic biosynthesis, some members, such as OleP (CYP107D1) and CYP107X1, have also been shown to catalyze steroid hydroxylation, albeit with limited substrate scope. In this study, we identified CYP107E41 from <em>Streptoalloteichus</em> sp. KCCM40925 as a novel member of the CYP107 family with selective hydroxylation activity toward both androstane and pregnane steroids. <em>In-vitro</em> screening using a panel of eleven steroids revealed predominant 6β- and 16α-hydroxylation, with androstane substrates exhibiting higher conversion efficiencies. Among the redox systems evaluated, diacetoxyiodobenzene provided the highest catalytic efficiency. Molecular docking identified two major binding orientations, C6–Fe and C16–Fe, with the C6–Fe configuration consistently scoring higher in CNN pose evaluations, suggesting it as the catalytically preferred binding mode. Molecular dynamics simulations further supported this, showing that the testosterone complex maintained a stable conformation with C6 positioned near the heme iron, consistent with selective mono-hydroxylation. In contrast, the nandrolone complex exhibited greater conformational flexibility, maintaining the proximity of both C6 and C7 to the heme, aligning with its broader hydroxylation profile. These findings establish CYP107E41 as a promising candidate for regioselective steroid hydroxylation and provide mechanistic insights to inform future protein engineering and redox system optimization.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110595"},"PeriodicalIF":3.0,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144863140","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"NNMT enhances invasive and migratory capacity of keloid fibroblasts and M2 macrophage polarization","authors":"Chengxiang Yan,&nbsp;Haijiang Dong,&nbsp;Zhibin Li,&nbsp;Xinhao Cheng,&nbsp;Ximei Wang","doi":"10.1016/j.abb.2025.110593","DOIUrl":"10.1016/j.abb.2025.110593","url":null,"abstract":"<div><div>Keloid are characterized by fibroblastic proliferation and excessive collagen deposition. Nicotinamide N-methyltransferase (NNMT) belongs to the methyltransferase family and plays an important role in various physiological processes, including keloid formation. However, the role of NNMT in keloid development remains poorly understood. In this study, we investigated the role of NNMT in keloids using a bleomycin (BLM)-induced fibrosis model in vivo and primary human keloid fibroblast (HPKF) in vitro. NNMT expression was upregulated in keloid tissues, and M2 macrophages were significantly increased in keloid tissue. NNMT overexpression reduced the SAM content, while NNMT knockdown increased it. Moreover, overexpression of NNMT enhanced the cell viability of HPKFs, while knockdown of NNMT promoted apoptosis. Additionally, NNMT overexpression promoted HPKF invasion and migration, whereas knockdown effectively inhibited these processes. Overexpression of NNMT also increased the expression of fibrosis-related markers, including FN1, COL1A1, COL3A1, Vimentin, and α-SMA, while NNMT knockdown reversed it. THP-1 cells were induced into THP1-M0 macrophages using PMA and co-cultured with HPKFs. Co-culturing with NNMT-overexpressing HPKFs promoted M2 polarization in M0 cells, while NNMT knockdown inhibited M2 polarization and reduced TGFB1 and Arg-1 expression in M0 cells. Furthermore, NNMT overexpression promoted IGF-1 expression, and treatment with AG1024 (an IGF1R inhibitor) suppressed NNMT-induced M2 polarization and inhibited IGF1B and Arg-1 expression. Taken together, our findings suggest that NNMT enhances the invasive and migratory capacity of keloid fibroblasts and regulates M2 macrophage polarization in keloids through IGF-1 modulation. These results highlight NNMT as a potential therapeutic target for keloid treatment.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110593"},"PeriodicalIF":3.0,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144862039","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular and structural characterization of ureido-benzenesulfonamides as dual inhibitors of aldose reductase and cholinesterases","authors":"Cüneyt Türkeş ,&nbsp;Nebih Lolak ,&nbsp;Hatice Esra Duran ,&nbsp;Gönül Yapar ,&nbsp;Suleyman Akocak","doi":"10.1016/j.abb.2025.110594","DOIUrl":"10.1016/j.abb.2025.110594","url":null,"abstract":"<div><div>The conformational and kinetic intricacies of enzyme-ligand interactions are critical for understanding molecular mechanisms underlying metabolic and neurodegenerative pathologies. Herein, we report a rationally designed series of ureido-benzenesulfonamides as dual-acting nanomolar inhibitors targeting aldose reductase (ALR2) and cholinesterases (AChE and BChE). Spectrophotometric inhibition assays revealed that compound <strong>3SA-a</strong> displayed exceptional ALR2 affinity (<em>K</em>_I = 7.00 ± 0.68 nM), surpassing epalrestat by over 30-fold. Likewise, <strong>3SA-f</strong> selectively inhibited BChE with a <em>K</em>_I of 24.20 ± 2.26 nM, outperforming tacrine by a factor of 7.8. Molecular docking simulations highlighted distinct dynamic binding modes: <strong>3SA-a</strong> engaged ALR2's catalytic cleft through a hydrogen bond with Tyr48 and water-bridged interactions, whereas <strong>3SA-f</strong> leveraged π-π stacking and halogen bonding within BChE's extended acyl pocket. These binding orientations were consistent with SAR findings, where meta-sulfonamide placement and halogen substitution optimized selectivity and conformational complementarity. Complementary <em>in silico</em> ADME-Tox predictions confirmed the drug-like nature of all compounds (0 Lipinski/PAINS violations), moderate oral permeability (QPPCaco: 79–85 nm/s), and low CNS exposure (CNS score = −2), aligning with a peripheral mechanism of action. Collectively, this study provides a detailed structural and dynamic framework for dual-target enzyme inhibition, offering a tunable scaffold for future therapeutics targeting the ALR2-ChE axis.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110594"},"PeriodicalIF":3.0,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144862038","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}