Biochemical pharmacology最新文献

{"title":"Comprehensive functional characterization of rare and known CYP2E1 allelic variants identified in a Japanese population.","authors":"Yuki Ohmori, Eiji Hishinuma, Yuma Suzuki, Akiko Ueda, Caroline Mwendwa Kijogi, Tomoki Nakayoshi, Akifumi Oda, Sakae Saito, Shu Tadaka, Kengo Kinoshita, Yu Sato, Masahiro Hiratsuka","doi":"10.1016/j.bcp.2025.117396","DOIUrl":"https://doi.org/10.1016/j.bcp.2025.117396","url":null,"abstract":"<p><p>Cytochrome P450 2E1 (CYP2E1) is a key hepatic enzyme involved in the oxidative metabolism of low-molecular-weight xenobiotics, including drugs, anesthetics, and procarcinogens. Although numerous CYP2E1 genetic variants have been identified, their functional implications remain unclear. In this study, we systematically characterized 25 CYP2E1 variants-22 novel missense variants identified via whole-genome sequencing of 8,380 Japanese individuals, and three PharmVar-defined alleles (*2, *3, and *4). The wild-type and variant enzymes were expressed in 293FT cells, and protein levels were quantified by Western blotting. Enzyme activity was evaluated by measuring chlorzoxazone 6-hydroxylation, and kinetic parameters (Km, Vmax, CLint) were derived from Michaelis-Menten analysis. Five variants exhibited significantly reduced catalytic activity, while two showed increased function. One frameshift variant, Leu447fs, resulted in complete loss of activity. In silico 3D docking simulations using a homology-modeled CYP2E1 structure revealed that several function-altering variants were located near the heme-binding domain or substrate recognition sites. Structural analysis of the Leu133His variant suggested that disruption of hydrogen bonding networks near the heme could underlie its reduced activity. The three known star alleles showed no significant deviation from wild-type activity. These findings provide important mechanistic insights into the functional consequences of rare CYP2E1 variants and underscore the enzyme's structural resilience. Our results offer foundational data for interpreting CYP2E1 genetic variation and support future efforts in personalized medicine through pharmacogenomic profiling.</p>","PeriodicalId":8806,"journal":{"name":"Biochemical pharmacology","volume":" ","pages":"117396"},"PeriodicalIF":5.6,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145231419","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dual targeting of EphAs and KDR axis hampers VEGF-induced angiogenesis and glioma stem cell replication.","authors":"Alfonso Zappia, Francesca Romana Ferrari, Carmine Giorgio, Stefano Sala, Ilaria Zanotti, Simona Parrinello, Melanie P Clements, Marco Rusnati, Michela Corsini, Andrea Blesio, Riccardo Castelli, Lorenzo Guidetti, Laura Scalvini, Alessio Lodola, Massimiliano Tognolini","doi":"10.1016/j.bcp.2025.117398","DOIUrl":"https://doi.org/10.1016/j.bcp.2025.117398","url":null,"abstract":"<p><p>Glioblastoma multiforme (GBM) is an aggressive and highly vascularized brain tumor with a poor prognosis and limited therapeutic options. Resistance to current treatments is largely driven by glioma stem-like cells (GSCs), a subpopulation with high tumorigenic potential that plays a key role in tumor progression, recurrence, and angiogenesis. Eph receptor tyrosine kinases, including EphA and EphB, are broadly implicated in GBM biology. While both classes contribute to tumor development and plasticity, EphA receptors are more directly involved in GSC maintenance and in crosstalk with the VEGF/VEGFR-2 axis, whereas EphB receptor dysregulation may promote tumor invasion. This subclass distinction makes selective targeting of EphA receptors an attractive therapeutic strategy. Here, we present the characterization of UniPR1449, a novel small-molecule selective EphA receptor inhibitor. UniPR1449 is a protein-protein interaction inhibitor (PPI-i) that blocks ephrin-A1-induced EphA2 phosphorylation, internalization, and degradation in GBM cell lines. In patient-derived GSCs, the compound significantly reduced proliferation and S-phase entry. Additionally, UniPR1449 impaired VEGF-induced angiogenesis in the chick chorioallantoic membrane (CAM) assay, while leaving FGF2-mediated vascularization unaffected. This dual mechanism of action-targeting both EphA signaling and VEGFR-2-mediated angiogenesis-highlights its therapeutic potential in addressing two key pathological features of GBM: vascular support and stem-like tumor cell renewal. Moreover, its selectivity for EphA receptors may offer a safety advantage over pan-Eph inhibitors, which could disrupt physiological EphB functions. Together, these results position UniPR1449 as a promising lead compound for the development of multitarget therapies against GBM.</p>","PeriodicalId":8806,"journal":{"name":"Biochemical pharmacology","volume":" ","pages":"117398"},"PeriodicalIF":5.6,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145231470","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pharmacological inhibition of JNK-MAPK disrupts cigarette smoke-induced RUNX2/Galectin-3 driven EMT and cancer stemness in lung adenocarcinoma.","authors":"Karan Chhetri, Jiten R Sharma, Rajesh Vasita, Rana P Singh, Umesh C S Yadav","doi":"10.1016/j.bcp.2025.117399","DOIUrl":"https://doi.org/10.1016/j.bcp.2025.117399","url":null,"abstract":"<p><p>Cigarette smoke (CS), a major driver of lung cancer (LC), promotes epithelial-mesenchymal transition (EMT) and stemness resulting in metastasis, therapy resistance, and recurrence, but the precise mechanism is elusive. Building on our earlier identification of Runt related transcription factor-2 (RUNX-2) and Galectin-3 (Gal-3) as mediators of CS-induced EMT, in this study, we aim to identify a potential molecular mechanism and delineate the upstream regulators of RUNX-2 using A549 lung adenocarcinoma cells and human small airway epithelial cells (SAECs) cultured at the air-liquid interface (ALI). CSE exposure markedly elevated intracellular reactive oxygen species (ROS), assessed via DCFH-DA assay, and promoted invasive behavior (Boyden chamber assay), spheroid formation, and colony formation, the hallmarks of cancer stemness. Expression analysis via RT-qPCR, immunoblotting, and immunocytochemistry revealed that CSE upregulated EMT and stemness-associated markers, notably via upregulating RUNX2 and Galectin-3, at both transcriptional and translational levels through the involvement of c-Jun N-terminal kinase- Mitogen-Activated Protein Kinase (JNK-MAPK) pathways. A specific pharmacological inhibitor of JNK (SP600125) significantly attenuated CSE-induced RUNX2 and Galectin-3 expression, and also reversed CSE-driven EMT marker alterations, suppressed transcriptional EMT perturbations, and reduced proinflammatory cytokines, including monocyte chemoattractant protein-1 (MCP-1), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-α). In conclusion, this study identifies that ROS/JNK/RUNX2/Gal-3 axis drives CS-induced oncogenic plasticity, suggesting that targeted inhibition of this pathway could be an effective strategy for mitigating CS-related LC progression.</p>","PeriodicalId":8806,"journal":{"name":"Biochemical pharmacology","volume":" ","pages":"117399"},"PeriodicalIF":5.6,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145231556","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"GA-017 attenuates OA by activating YAP/TAZ.","authors":"Xinhuo Li, Mingyang Lei, Qiannan Ding, Yangjun Xu, Xuanyuan Lu, Jiewen Zheng, Zhuolin Chen, Xuewen Liu, Xi Chen, Weiqi Han, Wei He, Wanglei Yang, Yu Qian","doi":"10.1016/j.bcp.2025.117389","DOIUrl":"https://doi.org/10.1016/j.bcp.2025.117389","url":null,"abstract":"<p><p>Osteoarthritis (OA) is a common disease that causes joint pain, mobility issues, and functional impairment in middle-aged and older individuals. However, effective medications for its treatment are lacking. YAP (Yes-associated protein) /TAZ (transcriptional co-activator with PDZ-binding motif) are important protective molecules that regulate the development of OA, and activating YAP/TAZ may be a potential treatment strategy. GA-017 is an inhibitor of LATS (large tumor suppressor kinase), an upstream molecule of YAP/TAZ in the Hippo signaling pathway, and has the potential to activate YAP/TAZ. However, whether GA-017 regulates chondrocyte metabolism and arthritis progression through YAP/TAZ activation remains unclear. In this study, we found that GA-017 activated YAP/TAZ and inhibited NF-κB (nuclear factor kappaB) signaling, thereby regulating anabolic and catabolic cell processes and inflammatory responses, ultimately suppressing OA in mouse and human knee cartilage. These results demonstrate that GA-017's activation of YAP/TAZ has significant potential in the treatment of OA and further suggest that drug-induced activation of YAP/TAZ is a feasible approach for treating OA.</p>","PeriodicalId":8806,"journal":{"name":"Biochemical pharmacology","volume":" ","pages":"117389"},"PeriodicalIF":5.6,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145231504","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Plectin promotes bone formation via phase separation and sequestering annexin A2.","authors":"Kaiyuan Zheng, Xi He, Yang Yu, Meng Deng, Lulu Chen, Wenwen Zhou, Dan Xia, Peihong Su, Ye Tian, Lijuan Zhang, Jing Wen, Beilei Zeng, Xiaolan Guo, Qi Liang, Bin Guo, Guangrong Wang, Yinxu Wang, Qing Wu, Li Jiao, Airong Qian, Chong Yin","doi":"10.1016/j.bcp.2025.117403","DOIUrl":"https://doi.org/10.1016/j.bcp.2025.117403","url":null,"abstract":"<p><p>Osteoporosis constitutes an emergent threat to human health, which partially caused by impaired osteoblast differentiation. Liquid-liquid phase separation (LLPS) is a cytophysiological process that regulates osteoblast differentiation. The plakin family, ubiquitously expressed in osteoblasts, interacts with multiple proteins and may mediate phase separation. However, limited reports exist regarding the involvement of the plakin family phase separation and osteoblast differentiation. In this study, we demonstrated that plectin (Plec), a plakin family member, enhances osteoblast differentiation and bone formation via its phase separation. This process is mediated by plectin's intrinsically disordered region (IDR). Mechanistically, plectin promotes osteoblast differentiation by sequestering annexin A2 (Anxa2), an osteoblast differentiation promoter, through phase separation. Furthermore, the functional amino acid region (aa 1967-2185) of plectin exhibited therapeutic effect in mice with osteoporosis. Our findings reveal a novel regulatory mechanism by which cytoskeletal linker proteins govern osteoblast differentiation and propose a potential therapeutic strategy for osteoporosis.</p>","PeriodicalId":8806,"journal":{"name":"Biochemical pharmacology","volume":" ","pages":"117403"},"PeriodicalIF":5.6,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145231545","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synaptic scaffold protein PSD-95: a therapeutic target for Alzheimer's disease.","authors":"Xing Fan, Hao Wang, Jianchao Ping, Mingzhe Li, Jianlan Gu, Wei Qian","doi":"10.1016/j.bcp.2025.117401","DOIUrl":"https://doi.org/10.1016/j.bcp.2025.117401","url":null,"abstract":"<p><p>Alzheimer's disease (AD) is a chronic neurodegenerative disorder marked by gradual cognitive deterioration and distinct neuropathological characteristics. The abnormal accumulation of amyloid-β (Aβ) and neurofibrillary tangles (NFTs) are the hallmarks of AD. In fact, synaptic loss and damage occur earlier than amyloid plaques and NFTs in the progression of AD and are most closely associated with the cognitive deficits exhibited by AD patients. In this review, we discuss the expression level, localization, posttranslational modification and interaction proteins of PSD-95, as well as their roles in synaptic plastisity. We also review the mechanisms through which PSD-95 contributes to synaptic dysfunction in AD. Moreover, the potential of PSD-95 as an early biomarker for AD is also discussed, along with the therapeutic approaches that target PSD-95 for patients afflicted with the disease. The objective of this review is to offer comprehensive insights into the early pathogenesis of Alzheimer's disease and to aid in the development of novel diagnostic and treatment methodologies grounded in this understanding.</p>","PeriodicalId":8806,"journal":{"name":"Biochemical pharmacology","volume":" ","pages":"117401"},"PeriodicalIF":5.6,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145231612","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genetic variants of cytochrome P450 2C21 identified by screening 6344 dogs influenced oxidations of the probe drug omeprazole","authors":"Yasuhiro Uno ,&nbsp;Koya Fukunaga ,&nbsp;Genki Ushirozako ,&nbsp;Norie Murayama ,&nbsp;Keijiro Mizukami ,&nbsp;Tomomi Aoi ,&nbsp;Hirotaka Tomiyasu ,&nbsp;Muneki Honnami ,&nbsp;Hajime Tsujimoto ,&nbsp;Masahiro Sakaguchi ,&nbsp;Masaharu Hisasue ,&nbsp;Taisei Mushiroda ,&nbsp;Yukihide Momozawa ,&nbsp;Hiroshi Yamazaki","doi":"10.1016/j.bcp.2025.117394","DOIUrl":"10.1016/j.bcp.2025.117394","url":null,"abstract":"<div><div>The cytochromes P450 (P450s or CYPs) are essential drug-metabolizing enzymes. In humans, the variability in metabolic activity is partly accounted for by genetic variants of the responsible P450. However, P450 genetic variants remain largely to be investigated in dogs, a species often used in drug metabolism studies. In this study, the sequencing of 6344 dog genomes found five <em>CYP2C21</em> variants, including three missense variants (p.K119Q, p.R125C, and p.R329H), one synonymous variant, and one frameshift variant (c.669dupA). The latter creates a premature termination codon that results in the elimination of the heme-binding region and substrate recognition sites 3–6; these are important functional domains, and thus c.669dupA is considered to be a null allele. Some of these variants were found only in a limited number of dogs and breeds, indicating that their distribution may be restricted. Recombinant proteins of the missense variants, co-expressed with NADPH-P450 reductase, were prepared in <em>Escherichia coli</em> for metabolic assays. The CYP2C21 R125C variant showed decreased metabolic capacity compared with the wild-type CYP2C21 protein. These results suggest the possible contribution of genetic variants of <em>CYP2C21</em> to the variability of drug metabolism in dogs.</div></div>","PeriodicalId":8806,"journal":{"name":"Biochemical pharmacology","volume":"242 ","pages":"Article 117394"},"PeriodicalIF":5.6,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145231463","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel phosphodiesterase 5 inhibitor, CPD1, alleviates liver fibrosis by modulating the Akt/NF-κB pathway to inhibit activated hepatic stellate cells.","authors":"Wenbin Feng, Jianqin Yang, Jiaxiu Lei, Limei Li, Changfeng Shan, Shenjie Chen, Zongmeng Zhang, Zhenggang Zhao, Sujin Zhou, Allan Zijian Zhao, Yunping Mu, Fanghong Li","doi":"10.1016/j.bcp.2025.117388","DOIUrl":"https://doi.org/10.1016/j.bcp.2025.117388","url":null,"abstract":"<p><p>Hepatic fibrosis is a progressive disorder marked by abnormal extracellular matrix buildup, with no effective antifibrotic drugs currently available. Recent evidence indicates that inhibiting phosphodiesterase 5 (PDE5) can have significant benefits for fibrotic diseases, suggesting PDE5 inhibitors may be effective antifibrotic agents. This study aimed to develop a new PDE5 inhibitor, potassium salt crystal form B (CPD1), which has much greater aqueous solubility than sildenafil. We assessed CPD1's efficacy in inhibiting hepatic stellate cells (HSCs) activation and investigated its mechanism of action. The therapeutic effect of CPD1 was studied in a carbon tetrachloride-induced liver fibrosis model, exploring its antifibrotic mechanisms via cyclic guanosine monophosphate (cGMP)-dependent protein kinase (PKG1) or Akt inhibitors and PKG1 overexpression in LX-2 cells. As anticipated, the expression of PDE5A was significantly elevated in both human and mouse fibrotic liver tissues, as well as in LX-2 cells induced by transforming growth factor-beta 1 (TGFβ1). In vivo, CPD1 reduced serum transaminases in a dose-dependent manner, mitigated liver damage, decreased collagen deposition, and suppressed the activation of HSCs. Additionally, CPD1 is more effective than sildenafil at a lower dosage. In vitro, CPD1 inhibited TGFβ1-induced activation of LX-2 and reduced the expression of fibrotic marker proteins and genes. Notably, the anti-fibrotic effects of CPD1 were completely negated following the administration of a PKG1 inhibitor. Mechanistically, the CPD1 intervention effectively countered the TGFβ1-induced increase in p-IκBα and p-P65. This study demonstrated that CPD1 mitigates liver fibrosis by activating the cGMP/PKG pathway, which in turn inhibits the AKT/NF-κB pathway. Therefore, it may be considered a potential therapeutic agent that warrants further investigation for the treatment of liver fibrosis.</p>","PeriodicalId":8806,"journal":{"name":"Biochemical pharmacology","volume":" ","pages":"117388"},"PeriodicalIF":5.6,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145231477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Defining the role of 12-lipoxygenase in regulating platelet-derived extracellular vesicles.","authors":"Alexis J Matthew, Jaël D Richard, Vanessa L Gauvin, Mathieu P A Hébert, Mohamed Touaibia, Eric P Allain, Luc H Boudreau","doi":"10.1016/j.bcp.2025.117404","DOIUrl":"https://doi.org/10.1016/j.bcp.2025.117404","url":null,"abstract":"<p><p>Platelets are traditionally recognized for their role in hemostasis and wound repair, yet they also play a pivotal role in intercellular communication through the release of platelet-derived extracellular vesicles (PMVs). These vesicles contribute to diverse physiological and pathological processes, but the mechanisms governing their biogenesis remain incompletely understood. One candidate regulator is 12-lipoxygenase (12-LO), an enzyme that metabolizes arachidonic acid into the lipid mediator 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE). While 12-LO has been linked to coagulation and neutrophil interactions, its involvement in PMV production has not been clearly defined. In this study, we investigated the role of 12-LO in PMV biogenesis using both human and murine platelet models. Pharmacological inhibition of 12-LO significantly reduced 12(S)-HETE levels and PMV release in activated human platelets. Similarly, platelets from 12-LO-deficient (Alox12^-/^-) mice exhibited markedly impaired PMV production upon stimulation. Subpopulation analyses revealed agonist-specific effects of 12-LO deficiency on distinct PMV subsets, including mitochondrial-containing vesicles. Importantly, supplementation with exogenous 12(S)-HETE partially restored PMV production in Alox12^-/^- platelets, confirming the functional relevance of this lipid mediator. Together, these findings identify 12-LO as a critical regulator of PMV biogenesis and suggest that the 12-LO/12(S)-HETE axis represents a novel therapeutic target in inflammatory and thrombotic disorders.</p>","PeriodicalId":8806,"journal":{"name":"Biochemical pharmacology","volume":" ","pages":"117404"},"PeriodicalIF":5.6,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145231501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Theaflavin reduces Achilles tendon heterotopic ossification in mice through the TGF-β/Smad signaling pathway.","authors":"Yuan Li, Chenglong Li, Wensheng Zhang, Shiyong Le, Jiongjiong Zhou, Bijun Luo, Pusheng Xie, Bo Yan","doi":"10.1016/j.bcp.2025.117387","DOIUrl":"https://doi.org/10.1016/j.bcp.2025.117387","url":null,"abstract":"<p><p>Heterotopic ossification (HO) affects millions of people worldwide. TGF-β/Smad signaling pathway plays an essential role in HO of the Achilles tendon. Recent studies have found that Theaflavin can regulate the TGF-β/Smad signaling pathway, suggesting Theaflavin may have a positive effect on HO. In this study, we aimed to study the effects of Theaflavin on preventing endochondral differentiation of Tendon-derived stem cells (TDSCs) and the HO process after the achilles tendon injury model. Here, we investigated the role of Theaflavin in a mouse model of Achilles tendon heterotopic ossification. In addition, we use TDSCs to explore the molecular mechanism of Theaflavin affecting HO. Our data showed that Theaflavin can inhibit HO of the Achilles tendon and significantly reduce the volume of mature bone tissues. SOX9 and RUNX2 were decreased significantly after Theaflavin administration. Experiments showed Theaflavin inhibited TGF-β/Smad signaling pathway during chondrogenic differentiation and osteogenic differentiation of TDSCs. Surface Plasmon Resonance Assay and Molecular docking simulation showed that a direct molecular interaction between Theaflavin and TβRI (a membrane receptor of TGF-βs). Meanwhile, Cellular Thermal Shift Assay showed Theaflavin bonded and thermally stabilized TβRI significantly. Moreover, WB and IHC displayed Theaflavin also exhibited inhibitory effects on TβRI phosphorylation. In summary, Our findings demonstrated that TF inhibited HO by down-regulating the TGF-β/Smad signal pathway, and this effect may attributed to TF binded directly to TβRI and prevented its phosphorylation.</p>","PeriodicalId":8806,"journal":{"name":"Biochemical pharmacology","volume":" ","pages":"117387"},"PeriodicalIF":5.6,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145231519","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}