Molecular Pharmacology最新文献

{"title":"Biasing G protein βγ subunit downstream signaling enhances the analgesic effects of endogenous opioid receptor agonists during nitroglycerin-induced thermal hypersensitivity.","authors":"Farzanna A Mohamed, Alan V Smrcka, Emily M Jutkiewicz","doi":"10.1016/j.molpha.2025.100068","DOIUrl":"10.1016/j.molpha.2025.100068","url":null,"abstract":"<p><p>μ-Opioid receptor (MOR) agonists are a mainstay in acute pain management. However, they also produce adverse effects and are frequently misused, increasing susceptibility for opioid use disorder. Thus, a strategy for improving the safety of opioid analgesics is needed. Gallein-mediated inhibition of Gβγ signaling to a subset of inhibitory effector feedback systems, such as phospholipase C (PLC) β3 and G protein-coupled receptor kinase 2, potentiates the antinociceptive effects of morphine without altering its rewarding effects in vivo. In this study, we examined effectiveness of gallein in the context of persistent pain using a nitroglycerin (NTG)-induced thermal hypersensitivity assay. In the warm water tail withdrawal assay, NTG (10 mg/kg, i.p.) decreased tail withdrawal latencies by 90%, from 30 to 3 seconds, indicating a hyperalgesic state. Gallein alone fully reversed NTG-induced decreases in withdrawal latencies. Furthermore, a low dose of gallein (3.2 mg/kg, i.p.) that was ineffective alone was sufficient to potentiate the antihyperalgesic effects of morphine. Pretreatment with the nonselective opioid antagonist naloxone (1.0 mg/kg, i.p.) attenuated both the antihyperalgesic effects of gallein alone and gallein-mediated potentiation of morphine. NTG did not decrease tail withdrawal latencies in PLCβ3^-/- mice compared with wild-type littermates, and this apparent antihyperalgesia was also reversed by naloxone. Taken together, the Gβγ inhibitor gallein alone produced antihyperalgesic effects mediated by endogenous opioid receptor activation. These data suggest that inhibiting Gβγ effectors, such as PLCβ3, downstream of MOR activation improves the analgesic effects of both endogenous and exogenous MOR agonists. SIGNIFICANCE STATEMENT: This study demonstrates the potential of small molecule-mediated modulation of Gβγ signaling to enhance the analgesic effects of endogenous opioid peptides suggesting a novel strategy for safer pain management, reducing the risk of opioid use disorder while maintaining effective pain relief.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"107 9","pages":"100068"},"PeriodicalIF":3.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12597591/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145008307","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabotropic glutamate receptor homo- and heterodimers exhibit distinct responses to orthosteric and allosteric ligands.","authors":"Tyler W McCullock, Tyler Couch, Paul J Kammermeier","doi":"10.1016/j.molpha.2025.100063","DOIUrl":"10.1016/j.molpha.2025.100063","url":null,"abstract":"<p><p>Metabotropic glutamate receptors (mGlus) are obligate dimer G protein-coupled receptors that can all homodimerize and heterodimerize in select combinations. Responses of mGlu heterodimers to selective ligands, including orthosteric agonists and allosteric modulators, are largely unknown. The pharmacological properties of each group II and III mGlu homodimer (except the exclusively retinally expressed mGlu6) and several heterodimers were examined when stochastically assembled in HEK293T cells, or specifically measured using an improved G protein-mediated bioluminescence resonance energy transfer assay employing complemented fragments of Nanoluciferase. Stochastically assembled receptors adopted unique signaling characteristics. Some favored the potency, efficacy, or signaling kinetics of a dominant subunit, whereas others exhibited blended profiles reflective of a combination of homo- and heterodimers at various ratios of expressed receptor. Finally, group II and III mGlu dimers were examined for responses to selective agonists and allosteric modulators. Effects of glutamate and selective group II and III orthosteric agonists were found to result in unique concentration response profiles when examining each combination of group II and III mGlu. Effects of select allosteric modulators were examined for each mGlu2-containing dimer as well as several group III dimer pairs. Likewise, allosteric modulator effects were often unique across dimers containing the targeted subunit of the ligand being tested. The results demonstrate that mGlu dimers respond uniquely to ligands selective for just one subunit, even when they are ostensibly pharmacologically similar. SIGNIFICANCE STATEMENT: This study demonstrates novel pharmacological responses to ligands acting on metabotropic glutamate receptor heterodimers in isolation. To our knowledge, it also shows for the first time the kinetics and pharmacological properties of group II and III metabotropic glutamate receptors when expressed in pairs and assembled stochastically.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"107 9","pages":"100063"},"PeriodicalIF":3.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12597640/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144883168","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Heterogeneous nuclear ribonucleoprotein A/B drives gastric cancer and epithelial-mesenchymal transition via the Akt-GSK3β-Wnt pathway.","authors":"Luo Huiru, Aime Gael Yaya Traore, Junyi Hu, Yinshuang Miao, Liu Yu, Mengzhen Huang, Zhongxue Guo, Qing Zheng, Feng Wang","doi":"10.1016/j.molpha.2025.100066","DOIUrl":"10.1016/j.molpha.2025.100066","url":null,"abstract":"<p><p>Gastric cancer (GC) is a leading cause of cancer-related deaths globally, with metastasis critically impacting prognosis. Splicing factors are key regulators of tumorigenesis, particularly in metastasis. In this exploratory study, we investigated the role and mechanism of heterogeneous nuclear ribonucleoprotein A/B (HNRNPAB) in GC cell invasion and migration. We detected a 3.45-fold increase in HNRNPAB protein levels in highly metastatic MKN45 cells compared with low metastatic MKN7 cells (P < .01) and marked upregulation in human GC tissues (n = 408) versus normal tissues (n = 211, P < .05, predicted by gene expression profiling interactive analysis). HNRNPAB overexpression in MKN45 and MKN7 cells substantially enhanced proliferation by 50% (P < .001, 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2-H-tetrazolium bromide assay), migration by 60% (P < .001, wound healing assay), and invasion by 70% (P < .001, Transwell assay), whereas knockdown reduced these by similar magnitudes. Mechanistically, HNRNPAB decreased E-cadherin (0.5 ± 0.1-fold, P < .001) and increased N-cadherin (1.8 ± 0.2-fold), Vimentin (2.0 ± 0.2-fold), and Snail levels (1.7 ± 0.2-fold, P < .001), promoting epithelial-mesenchymal transition, which was associated with Akt-GSK3β-Wnt pathway modulation by elevating phosphorylated Akt (Ser473, 2.0 ± 0.2-fold) and phosphorylated glycogen synthase kinase 3β (Ser9, 1.8 ± 0.2-fold, P < .001). These findings suggest that HNRNPAB is a potential diagnostic and therapeutic target for GC. SIGNIFICANCE STATEMENT: This exploratory study reveals that heterogeneous nuclear ribonucleoprotein A/B is associated with gastric cancer progression and epithelial-mesenchymal transition through its potential modulation of the Akt-GSK3β-Wnt signaling pathway. Targeting heterogeneous nuclear ribonucleoprotein A/B could offer novel therapeutic approaches for improving treatment outcomes in gastric cancer, highlighting its potential as a biomarker for disease prognosis.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"107 9","pages":"100066"},"PeriodicalIF":3.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12597541/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145008339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Interaction between W41 of the hepatitis B virus preS1 surface peptide and Y146/F274 of the cellular receptor molecule Na⁺/taurocholate co-transporting polypeptide is essential for virus entry.","authors":"Sebastian Kunz, Lena Soppa, Regina Leidolf, Anita Neubauer, Thomas Lütteke, Dieter Glebe, Joachim Geyer","doi":"10.1016/j.molpha.2025.100069","DOIUrl":"10.1016/j.molpha.2025.100069","url":null,"abstract":"<p><p>The myristoylated preS1 domain (myr-preS1) of the hepatitis B virus (HBV) large surface protein is essential for binding to the receptor protein, Na⁺/taurocholate co-transporting polypeptide (NTCP), and for the subsequent internalization of the virus-receptor complex. NTCP, which is expressed in hepatocytes, plays a physiological role in hepatic bile acid transport. Recent cryo-electron microscopy structures of the myr-preS1-NTCP complex were used to analyze virus-receptor interactions at the molecular level. Several interaction domains with high binding energies have been identified, including the interaction between myr-preS1 tryptophan 41 (W41) and the NTCP residues tyrosine 146 and phenylalanine 274 (Y146/F274), which are located at a considerable distance from the physiological bile acid binding sites of NTCP. The Y146A/F274A mutant of NTCP retained bile acid transport function but showed a reduced myr-preS1 binding. The W41G myr-preS1 mutant exhibited reduced binding to wild-type NTCP. The Y146A/F274A NTCP mutant did not support in vitro HBV infection, and the W41G myr-preS1 peptide was less effective in blocking infection compared with the wild-type myr-preS1 peptide. In conclusion, the myr-preS1-W41/NTCP-Y146/F274 interaction site, characterized by high binding energy, is essential for HBV entry into hepatocytes. Because this domain is spatially distinct from the bile acid binding and translocation sites of NTCP, it presents an attractive receptor target site for structure-based development of virus-selective HBV entry inhibitors that preserve the physiological bile acid transport function of NTCP. SIGNIFICANCE STATEMENT: This study identified and validated a high energy interaction site between the hepatitis B virus and its receptor Na⁺/taurocholate co-transporting polypeptide that can be used for structure-based drug design of virus entry inhibitors.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"107 9","pages":"100069"},"PeriodicalIF":3.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12597529/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145030300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Design and evaluation of highly selective histone deacetylase 6 inhibitors derived from the natural product tryptoline.","authors":"Khan Hashim Ali, Chiranjeev Sharma, Yong Jin Oh, Ji Hoon Yu, Soong-Hyun Kim, Heejin Lee, Young Ho Seo","doi":"10.1016/j.molpha.2025.100062","DOIUrl":"10.1016/j.molpha.2025.100062","url":null,"abstract":"<p><p>The development of isoform-selective histone deacetylase (HDAC) inhibitors offers a promising approach to minimize the adverse effects of nonselective HDAC inhibitors. HDAC6, due to its unique structural and functional properties, regulates critical cellular processes like gene expression, proliferation, senescence, and apoptosis. Inspired by a tryptoline-derived natural product, callophycin A, a series of compounds were synthesized and evaluated for HDAC6 selectivity. In the HDAC enzyme assay, compound 6a stood out as the lead, demonstrating 21-fold higher potency against HDAC6 compared with HDAC1 (HDAC6 IC₅₀ = 83.6 ± 1.1 nM vs HDAC1 IC₅₀ = 1790 ± 1.0 nM). In dose-dependent western blot experiments using H1975 lung cancer cells, a lower concentration of compound 6a (3 μM) induced significantly greater acetylation of α-tubulin compared with histone H3, indicating preferential inhibition of cytoplasmic HDAC6 over the nuclear HDAC1 isoform. Molecular docking of compound 6a at the HDAC6 active site (PDB code: 5EDU) revealed key interactions including π-alkyl contacts via the cap group, π-π stacking through the linker, hydrogen bonding involving the zinc-binding group, and Zn²⁺ chelation by the hydroxamic acid moiety that support its strong and selective binding, consistent with its HDAC6 inhibitory profile. Overall, compound 6a represents a promising prototype for the rational design of selective HDAC6 inhibitors, offering a structural framework for developing safer and more effective therapeutics aimed at HDAC6-driven cancers, thereby advancing targeted drug development in oncology. SIGNIFICANCE STATEMENT: Selective histone deacetylase 6 (HDAC6) inhibitors provide a safer alternative to nonselective HDAC inhibitors, with potential applications in cancer. This study identifies compound 6a as a promising lead with remarkable HDAC6 specificity, offering a foundation for developing targeted and efficient therapeutics.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"107 8","pages":"100062"},"PeriodicalIF":3.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144835772","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An overview of autophagy inhibition as a potential clinical strategy in cancer therapy.","authors":"Ahmed M Elshazly, Nayyerehalsadat Hosseini, Aya A Elzahed, David A Gewirtz","doi":"10.1016/j.molpha.2025.100056","DOIUrl":"10.1016/j.molpha.2025.100056","url":null,"abstract":"<p><p>Autophagy is a cellular process responsible for the recycling of misfolded proteins and damaged organelles, contributing to cellular homeostasis and energy production. Tumor cells often exploit this mechanism, particularly through the form of autophagy that is cytoprotective, to survive endogenous and exogenous stress and resist chemotherapeutic agents as well as radiation therapy. Although several autophagy inhibitors have been developed to block the protective form of autophagy, their clinical application is often limited due to a lack of selectivity and significant side effects. In addition to the cytoprotective form, cytotoxic, cytostatic, and nonprotective functions of autophagy have been identified. In this review, we summarize a series of publications, largely from our own laboratory, exploring how various antineoplastic agents trigger different forms of autophagy and assess whether autophagy inhibition or modulation could serve as an effective adjuvant approach to enhance therapeutic responses. Furthermore, we discuss recent advancements in the autophagy field and the potential for improving cancer therapeutic strategies. SIGNIFICANCE STATEMENT: This work provides an overview of our previous work investigating the different forms of autophagy induced by various antineoplastic modalities across different tumor models. The purpose of this effort is to draw tentative conclusions regarding the potential of targeting autophagy as a strategy to enhance the efficacy of these therapeutic agents. Additionally, we offer insights into recent advances in the autophagy field.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"107 8","pages":"100056"},"PeriodicalIF":3.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13632086/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144668016","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corticosterone stimulates synthesis of 2-arachidonoylglycerol via putative membrane-bound glucocorticoid receptors and inhibits GABA release via CB1 cannabinoid receptors in the ventrolateral periaqueductal gray.","authors":"Basile Coutens, Courtney A Bouchet, Lorenzo C Patti, Kylie B McPherson, Bethany S Boston, David C Jewett, Susan L Ingram","doi":"10.1016/j.molpha.2025.100058","DOIUrl":"10.1016/j.molpha.2025.100058","url":null,"abstract":"<p><p>The ventrolateral periaqueductal gray (vlPAG) plays a critical role in pain modulation. GABAergic neurotransmission within the vlPAG regulates the descending pain pathway. This study investigates the mechanisms through which corticosterone (CORT) modulates GABA release in the vlPAG via putative membrane-associated glucocorticoid receptors (mbGRs). Superfusion of CORT decreases evoked inhibitory postsynaptic currents in a mbGR- and CB1 cannabinoid receptor (CB1R)-dependent manner. Using a depolarization-induced suppression of inhibition protocol to test the effects of CORT on the endocannabinoid system, we find that CORT-mediated signaling enhances 2-arachidonoylglycerol synthesis that is inhibited by the diacylglycerol lipase inhibitor, DO34. CORT prolongs CB1R activation through a Gα_s and protein kinase A-dependent pathway, whereas early depolarization-induced suppression of inhibition-initiated endocannabinoid activation of CB1Rs is independent of protein kinase A. These results highlight the critical role of CORT in the vlPAG in engaging endocannabinoid pathways to inhibit GABA release. The results indicate that CORT activation of putative mbGRs promote activation of the descending pain modulatory pathway through CB1R-mediated inhibition of GABA release in the vlPAG. SIGNIFICANCE STATEMENT: This study provides evidence that corticosterone activates putative membrane glucocorticoid receptors to increase levels of 2-arachidonoylglycerol to activate presynaptic CB1 cannabinoid receptors. These findings reveal mechanisms by which stress modulates the ventrolateral periaqueductal gray and the descending pain circuit.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"107 8","pages":"100058"},"PeriodicalIF":3.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12402929/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144743153","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative evaluation of the biological characteristics of a novel retinoid X receptor agonist and bexarotene.","authors":"Koji Tomita, Ken-Ichi Nakashima, Eiji Yamaguchi, Akichika Itoh, Kaname Tsutsumiuchi, Makoto Inoue","doi":"10.1016/j.molpha.2025.100057","DOIUrl":"10.1016/j.molpha.2025.100057","url":null,"abstract":"<p><p>The retinoid X receptor (RXR) belongs to the nuclear receptor superfamily, which regulates various physiological processes. RXR agonists, classified as rexinoids, exhibit selectivity for RXR over the retinoic acid receptor and have therapeutic potential against cancer, metabolic disorders, and Alzheimer disease (AD). Here, we characterized the biological properties of 6-hydroxy-3'-propyl-[1,1'-biphenyl]-3-propanoic acid (6OHA), a compound synthesized in our laboratory based on the structure of magnaldehyde B, and found that it exhibited potent RXRα agonist activity comparable with that of the clinically used RXR agonist bexarotene, but lower agonist activity toward retinoic acid receptor α and RXRγ. RNA-sequencing-based transcriptome analysis of microglial cells revealed that 6OHA and Bex induced similar gene expression patterns; however, 6OHA was more associated strongly with chemotaxis and response to stimuli. Pharmacokinetic studies showed a higher C_max, faster T_max, and more rapid clearance in both the serum and brain for 6OHA than for Bex. Although 6OHA exhibited a higher area under the concentration-time curve from 0 to 6 hours in serum, its area under the concentration-time curve from 0 to 6 hours in the brain was lower than that of Bex. Together with gene expression data, these findings suggest that 6OHA is a more effective RXR agonist in peripheral tissues while maintaining comparable efficacy in the brain. Furthermore, unlike Bex, 6OHA did not increase serum triglycerides or decrease serum thyrotropin and free thyroxine levels, likely reflecting its distinct pharmacologic profile from that of Bex. Collectively, these results suggest that 6OHA is a promising RXR agonist with minimal adverse effects and potential application in treating cancer, AD, and metabolic disorders. SIGNIFICANCE STATEMENT: Despite the potential of retinoid X receptor (RXR) agonists, their application has been limited by adverse effects. The novel RXR agonist 6OHA exhibits potent RXRα agonist activity while showing lower activities for retinoid acid receptor α and RXRγ than the RXR agonist bexarotene. Moreover, 6OHA also shows favorable pharmacokinetics and a gene expression profile distinct from Bex. These properties may account for the minimal adverse effects of 6OHA and support its potential as a therapeutic RXR agonist.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"107 8","pages":"100057"},"PeriodicalIF":3.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144699042","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigation of the binding mode of clobenprobit at CXCR4 and development of novel anti-inflammatory compounds with enhanced activity and minimal antagonist effects.","authors":"Birgit Caspar, Nassima Bekaddour, Séverine Grinberg, Dominique Cathelin, Ivana Stoilova, Stephane Giorgiutti, Vincent Gies, Anne-Sophie Korganow, Nicolas Pietrancosta, Didier Roche, Vincent Rodeschini, Jean-Philippe Herbeuval","doi":"10.1016/j.molpha.2025.100055","DOIUrl":"10.1016/j.molpha.2025.100055","url":null,"abstract":"<p><p>IT1t and clobenprobit (CB) have been shown to act as anti-inflammatory compounds in dependence of the chemokine receptor C-X-C receptor type 4 (CXCR4) in model systems. Here, the direct interaction between CB and CXCR4 is demonstrated via in silico modeling and bioluminescence resonance energy transfer binding assays at wild-type and mutant versions of CXCR4. The binding site is compared with those of IT1t and AMD3100, two well known CXCR4 ligands. In contrast to AMD3100, IT1t also displays an anti-inflammatory signaling effect. Ligands observed to have this anti-inflammatory effect seem to bind into the minor pocket of CXCR4 impacting the binding of the endogenous ligand CXCL12 only at high concentrations. Based on this observation further compounds thought to bind the minor pocket of CXCR4 were designed and screened for their anti-inflammatory potency. The best of these compounds, NP1411, was tested in its ability to inhibit CXCL12 mediated G protein activation as well as CXCL12 and CB binding. SIGNIFICANCE STATEMENT: This study presents a comprehensive investigation into the binding site of anti-inflammatory compounds at the C-X-C receptor type 4 receptor using in silico and in vitro ligand binding approaches. This opens the opportunity for the development of further therapeutic agents with higher potency and/or efficacy as presented in an initial test at the end of the publication.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"107 8","pages":"100055"},"PeriodicalIF":3.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144663968","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Progress toward new function and design of extracellular G protein-coupled receptor nanobodies.","authors":"Roman R Schlimgen, Brian F Volkman","doi":"10.1016/j.molpha.2025.100054","DOIUrl":"10.1016/j.molpha.2025.100054","url":null,"abstract":"<p><p>Antibodies have played a pivotal role in G protein-coupled receptor (GPCR) research and drug development. Nanobodies, or variable domain heavy chain-only antibodies, have emerged as a next-generation antibody with unique advantages in targeting GPCRs. The first generation of intracellular nanobodies have been instrumental in stabilizing GPCR structures for crystallography and in enabling in vitro GPCR imaging. More recently, extracellular-targeted nanobodies have demonstrated diverse pharmacological profiles, with the ability to modulate GPCR activity, localization, and downstream signaling. With these newly uncovered functional properties, nanobodies can be viewed not only as structural tools but also as modulators of receptor pharmacology. We highlight recent innovations in extracellular GPCR-targeting nanobodies and assess several approaches to accelerate their development as versatile research tools and therapeutics. SIGNIFICANCE STATEMENT: Nanobodies have emerged as a next-generation antibody platform with distinct advantages for targeting G protein-coupled receptors. This review highlights recent advances in extracellular G protein-coupled receptor-targeting nanobodies and explores innovative strategies to accelerate their development as powerful research tools and therapeutic agents.</p>","PeriodicalId":18767,"journal":{"name":"Molecular Pharmacology","volume":"107 8","pages":"100054"},"PeriodicalIF":3.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13095440/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144690971","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}