ACS Pharmacology and Translational Science最新文献

{"title":"Development of a Ghrelin Deacylase to Attenuate Drug Reward and Associated Effects of Methamphetamine","authors":"Madeline J. Stewart,&nbsp;Huimei Wei,&nbsp;Nellore Bhanu Chandar,&nbsp;Fang Zheng* and Chang-Guo Zhan*,&nbsp;","doi":"10.1021/acsptsci.5c00391","DOIUrl":"https://doi.org/10.1021/acsptsci.5c00391","url":null,"abstract":"<p >All addictive substances directly or indirectly interact with the dopamine reward system to alter the brain’s dopamine receptor activities. It is essential for a truly effective addiction medication to attenuate substance reward and normalize the brain’s physiological functions. Conventional pharmacological intervention approaches to the treatment of substance addiction usually aim to develop and deliver a potential therapeutic agent to the brain to directly block or decrease actions of the substance or its therapeutic target in the brain. However, it is a grand challenge to attenuate the substance reward without affecting the normal physiological functions of brain receptors or transporters. Here, we show that peripheral ghrelin deacylation using a ghrelin deacylase identified in this study can effectively attenuate the pharmacological and rewarding effects of methamphetamine, a representative psychostimulant, in rodents through an interesting pharmacological mechanism without interacting with the ghrelin receptor or the dopamine receptor (because the ghrelin deacylase is not expected to cross the blood–brain barrier). In further animal behavioral studies, ghrelin deacylase administration significantly attenuated rat self-administration of methamphetamine, suggesting that ghrelin deacylase may serve as a promising therapeutic candidate for addiction treatment.</p>","PeriodicalId":36426,"journal":{"name":"ACS Pharmacology and Translational Science","volume":"8 9","pages":"3259–3268"},"PeriodicalIF":3.7,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036702","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Intrinsically Disordered Regions of Zinc Finger Protein 687 in Phase Separation: A Potentially Pathogenic Driver of Paget’s Disease of Bone","authors":"Jing Wang,&nbsp;Zheng Guo,&nbsp;Muqing Zhang and Hu Wang*,&nbsp;","doi":"10.1021/acsptsci.5c00467","DOIUrl":"https://doi.org/10.1021/acsptsci.5c00467","url":null,"abstract":"<p >Paget’s disease of bone (PDB) is a chronic skeletal disorder characterized by abnormal bone remodeling and structural deformities, yet its underlying molecular mechanisms remain poorly understood. Genetic mutations in regulatory genes have been implicated in the PDB, potentially disrupting osteoclast activity and leading to excessive bone resorption and abnormal bone formation. Current research has largely focused on clinical phenotypes and downstream pathways, relying on molecular biology approaches and omics analyses to elucidate mechanisms. However, the impact of genetic mutations on the biophysical properties and functional dynamics of biomacromolecules remain unclear. Phase separation (PS), a process essential for biomolecular condensate organization and cellular function, has been implicated in various diseases but is unexplored in the PDB. This study investigates the PS properties of ZNF687, a zinc finger protein associated with PDB. Structural predictions and sequence analyses identified intrinsically disordered regions (IDRs) within ZNF687 that may drive PS, while protein interaction predictions suggest its involvement in bone remodeling pathways. These findings provide a novel perspective linking PS to PDB pathogenesis and highlight ZNF687 as a potential target for further investigation.</p>","PeriodicalId":36426,"journal":{"name":"ACS Pharmacology and Translational Science","volume":"8 9","pages":"2855–2867"},"PeriodicalIF":3.7,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036700","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bioglass/Ceria Nanoparticle Hybrids for the Prophylactic Treatment of Seroma: A Comparative Short-Term Study in Rats","authors":"Michael-Alexander Pais*,&nbsp;Simone de Brot,&nbsp;Robert Nißler,&nbsp;Isabel Arenas Hoyos,&nbsp;Athanasios Papanikolaou,&nbsp;Davide Bottone,&nbsp;Alexander Gogos,&nbsp;Anja Helmer,&nbsp;Robert Rieben,&nbsp;Mihai Constantinescu,&nbsp;Tino Matter,&nbsp;Inge Herrmann and Ioana Lese,&nbsp;","doi":"10.1021/acsptsci.5c00327","DOIUrl":"https://doi.org/10.1021/acsptsci.5c00327","url":null,"abstract":"<p >Seroma formation remains a common postoperative complication. While optimal treatment remains unclear, recent attention has turned to bioglass/ceria nanoparticle (NP) treatment for seromas. Previous work showed complete seroma resolution in a rat model after NP treatment in the long term. This study aimed to assess the short-term prophylactic effects of NPs. Twenty male Lewis rats underwent bilateral seroma induction surgery. Postoperatively, seroma cavities were treated with NPs, vehicle buffer solution, or fibrin glue or left untreated. Over 2 weeks, blood, seroma fluid, and tissues were collected for biochemical, histopathological, and immunohistochemical analyses. By day 14, NP-treated seromas showed 100% fluid resolution. In contrast, seromas persisted in 50% of fibrin glue–treated rats, 60% of vehicle-treated rats, and 44.44% of untreated controls. Furthermore, prophylactic NP treatment resulted in decreased levels of inflammatory markers while the effect of fibrin glue was to increase the pro-inflammatory response. Histologically, a reduction in vascularization and individual macrophage infiltration was observed in seroma superficial capsules after NP treatment, while complement proteins were significantly increased and associated with groups of macrophages that colocalized with NPs. At the end point, NPs did not show any biodistribution to the systemic circulation. Prophylactic NP application reduced early seroma manifestations mostly through their anti-inflammatory effects. Members of the complement cascade were also identified in macrophages that colocalized with NPs and were internalized. Moreover, there were no detectable adverse systemic effects. These findings emphasize the clinical potential of NPs in the prevention of seromas and their potential for clinical use.</p>","PeriodicalId":36426,"journal":{"name":"ACS Pharmacology and Translational Science","volume":"8 9","pages":"3170–3181"},"PeriodicalIF":3.7,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acsptsci.5c00327","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Small-Molecule Activator of Sarco/Endoplasmic Reticulum Ca2+-ATPase Attenuates Cerebral Ischemia–Reperfusion Injury by Suppressing Endoplasmic Reticulum Stress and Apoptosis","authors":"Vikrant Rahi,&nbsp; and ,&nbsp;Ravinder K. Kaundal*,&nbsp;","doi":"10.1021/acsptsci.5c00151","DOIUrl":"https://doi.org/10.1021/acsptsci.5c00151","url":null,"abstract":"<p >Calcium (Ca²⁺) homeostasis is critical for neuronal survival and function, which is regulated by a network of Ca²⁺-handling proteins. Among these, the sarco/endoplasmic reticulum calcium ATPase (SERCA) pump, located on the SR/ER membrane, plays a pivotal role in sequestering Ca²⁺ into the ER, thereby maintaining low cytosolic Ca²⁺ levels. Dysregulated SERCA function during ischemia contributes to ER Ca²⁺ depletion, resulting in intracellular Ca²⁺ imbalance and ER stress, both of which are implicated in the pathogenesis of ischemic reperfusion injury; SERCA has thus emerged as a potential pharmacological target for ischemic stroke. CDN1163, a SERCA activator, has shown promising effects in preclinical studies of neurodegenerative diseases by alleviating ER stress and restoring the Ca²⁺ balance. This study investigates the neuroprotective potential of CDN1163 against cerebral ischemia-reperfusion (IR) injury using middle cerebral artery occlusion (MCAO) in rats. CDN1163 treatment (10 mg/kg, i.p.) significantly improved neurological scores, motor function, and behavior, while reducing infarct volume, brain edema, and oxidative stress by decreasing nitrite and lipid peroxidation and restoring glutathione levels. Histological analysis revealed reduced neuronal damage in the cortex, subcortex, and hippocampus regions. CDN1163 restored SERCA2b and 1a expression and mitigated ER stress by decreasing the expression of ER stress markers, such as PDI, BiP, p-IRE1α, XBP1, p-PERK, p-eIF2α, ATF4, and ATF6. Furthermore, CDN1163 downregulated pro-apoptotic markers Bax and CHOP, while upregulating the antiapoptotic protein Bcl-2 with TUNEL assay confirming decreased apoptosis. These outcomes highlight that CDN1163 is a potential therapeutic candidate for ischemic stroke, as it restores SERCA expression, alleviates endoplasmic reticulum stress, reduces oxidative stress, and inhibits apoptosis.</p>","PeriodicalId":36426,"journal":{"name":"ACS Pharmacology and Translational Science","volume":"8 9","pages":"2953–2963"},"PeriodicalIF":3.7,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036644","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nanomotion-Based Drug Sensitivity Prediction in Ovarian and Colon Cancer Cell Lines Using Machine Learning","authors":"Katja Fromm,&nbsp;Jan Winnicki,&nbsp;Grzegorz Jóźwiak,&nbsp;Gino Cathomen,&nbsp;Christine Wagner,&nbsp;Marta Pla Verge,&nbsp;Eric Delarze,&nbsp;Michał Świątkowski,&nbsp;Grzegorz Wielgoszewski,&nbsp;Maria Ines Villalba,&nbsp;Laura Munch,&nbsp;Sandor Kasas,&nbsp;Danuta Cichocka and Alexander Sturm*,&nbsp;","doi":"10.1021/acsptsci.5c00312","DOIUrl":"https://doi.org/10.1021/acsptsci.5c00312","url":null,"abstract":"<p >Cancer drug resistance remains a critical challenge in oncology, demanding rapid and reliable diagnostic tools to assess tumor cell susceptibility to treatment. This study presents a nanomotion-based drug susceptibility testing (DST) approach, integrating nanoscale movement analysis with supervised machine learning to classify drug-sensitive and drug-resistant cancer cells. Using label-free, real-time nanomotion technology, we measured the dynamic responses of colon cancer (SW480) and ovarian cancer (A2780, A2780ADR) cells to doxorubicin under physiological conditions. Features extracted from nanomotion signals were used to train machine learning models, achieving 90.9% accuracy in distinguishing between doxorubicin-treated and untreated SW480 cells and 84.6% accuracy in classifying doxorubicin-sensitive and -resistant ovarian cancer cells. The model achieved perfect classification of resistant A2780ADR cells in an independent test set after only 4 h and 15 min of exposure to the drug. Unlike genetic tests that infer drug resistance from molecular markers or metabolic assays requiring extended incubation times, nanomotion-based DST provides a direct phenotypic readout, offering a faster, label-free alternative for assessing tumor cell responses. While further dataset expansion and model refinement are necessary to enhance generalizability, these results underscore the potential of nanomotion technology as a rapid, phenotypic DST for personalized oncology. By directly measuring the mechanical behavior of cancer cells in response to chemotherapy, this method could transform clinical decision-making, enabling faster, more precise treatment strategies to combat drug resistance in cancer.</p>","PeriodicalId":36426,"journal":{"name":"ACS Pharmacology and Translational Science","volume":"8 9","pages":"3149–3159"},"PeriodicalIF":3.7,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acsptsci.5c00312","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036378","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DNA to RNA Tuning of a Pyrrolidine-Derived Quinoxaline Scaffold: Magnesium Induces Binding and Inhibition of Internal Ribosomal Entry Site RNA-Mediated Translational of Hepatitis C Virus","authors":"Rimita Saha,&nbsp;Bhim Majhi,&nbsp;Subhadeep Palit,&nbsp;Krishnamoorthi Srinivasan,&nbsp;Nilam Waghela,&nbsp;Dixit Tandel,&nbsp;Ankit Dhur,&nbsp;Rounak Patra,&nbsp;Mayank Gardia,&nbsp;Abhi Das,&nbsp;Dipendu Patra,&nbsp;Biswadip Chakraborty,&nbsp;Jayati Sengupta,&nbsp;Krishnan H Harshan,&nbsp;Mandar V. Deshmukh and Sanjay Dutta*,&nbsp;","doi":"10.1021/acsptsci.5c00160","DOIUrl":"https://doi.org/10.1021/acsptsci.5c00160","url":null,"abstract":"<p >Selective targeting of disease-relevant RNA with small molecules remains challenging due to the dynamic conformation of RNA and the nonspecific binding of ligands toward DNA. The internal ribosome entry site (IRES) RNA of the hepatitis C virus (HCV) is a critical antiviral drug target for structure-specific therapeutics. Small molecule inhibitors capable of altering the Mg²⁺ ion-induced L-shaped conformation of IRES subdomain IIa can effectively suppress the IRES-driven translation and HCV RNA replication. Here, we report the development of quinoxaline analogues designed to shift binding selectivity from DNA to HCV-IRES RNA by modifying a known DNA intercalator and describe their structure–activity relationship (SAR). Lead compound <b>8e</b>, featuring a pyrrolidine ring with an ether modification, exhibited Mg²⁺-dependent binding to HCV-IRES subdomain IIa RNA, evading DNA interactions and inhibiting HCV-IRES-mediated translation. Biophysical studies (CD, NMR, and FRET) revealed that <b>8e</b> undergoes a conformational change in the presence of Mg²⁺, favoring its binding to HCV-IRES RNA and altering its subdomain IIa structure, preventing the binding of the 40S ribosome. Moreover, <b>8e</b> significantly reduced the viral load in HCV-infected cells without observable cytotoxicity. Overall, our study highlights a strategy for fine-tuning quinoxaline-based small molecules to enhance RNA selectivity over DNA, and we present <b>8e</b> as a promising lead for HCV therapeutics.</p>","PeriodicalId":36426,"journal":{"name":"ACS Pharmacology and Translational Science","volume":"8 9","pages":"2964–2982"},"PeriodicalIF":3.7,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036234","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to: “Shifting the Antibody-Drug Conjugate Paradigm: A Trastuzumab-Gold-Based Conjugate Demonstrates High Efficacy Against Human Epidermal Growth Factor Receptor 2-Positive Breast Cancer Mouse Model”","authors":"Afruja Ahad,&nbsp;Hiwa K. Saeed,&nbsp;Virginia del Solar,&nbsp;Javier E. López-Hernández,&nbsp;Alexa Michel,&nbsp;Joshua Mathew,&nbsp;Jason S. Lewis* and Maria Contel*,&nbsp;","doi":"10.1021/acsptsci.5c00539","DOIUrl":"https://doi.org/10.1021/acsptsci.5c00539","url":null,"abstract":"","PeriodicalId":36426,"journal":{"name":"ACS Pharmacology and Translational Science","volume":"8 9","pages":"3389–3390"},"PeriodicalIF":3.7,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036207","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Highly Potent and Subtype-Selective Sperm-Specific Potassium Channel SLO3 Inhibitors Display High Tissue Exposure in the Murine Female Reproductive Tract","authors":"Kayla J. Temple*,&nbsp;Ping Li,&nbsp;Hallie G. McKinnie,&nbsp;Analisa Thompson Gray,&nbsp;Jeanette L. Bertron,&nbsp;Anna E. Ringuette,&nbsp;Pedro de Andrade Horn,&nbsp;Maximilian D. Lyon,&nbsp;Shweta Bhagwat,&nbsp;Leila Asadi,&nbsp;Sophia Li,&nbsp;Roman M. Lazarenko,&nbsp;Ronald McCarthy,&nbsp;Sichen Chang,&nbsp;Jeremy A. Turkett,&nbsp;Valerie Kramlinger,&nbsp;Katherine J. Watson,&nbsp;Irene A. Zagol-Ikapitte,&nbsp;Jerod S. Denton,&nbsp;Celia M. Santi,&nbsp;Carrie K. Jones,&nbsp;Craig W. Lindsley and Olivier Boutaud*,&nbsp;","doi":"10.1021/acsptsci.5c00416","DOIUrl":"https://doi.org/10.1021/acsptsci.5c00416","url":null,"abstract":"<p >To date, the study of drug exposure and distribution throughout the female reproductive tract has often been overlooked and neglected. Herein, we report the first highly potent and subtype-selective sperm-specific potassium channel 3 (SLO3) inhibitors as a new modality of contraceptives. After extensive <i>in vitro</i> and <i>in vivo</i> drug metabolism and pharmacokinetic profiling we selected <b>VU6032735</b> to utilize as a tool compound. <b>VU6032735</b> displayed low observed clearance with a long half-life and a high volume of distribution in female mice, which is seemingly an ideal target profile for a nonhormonal contraceptive. After intraperitoneal injection, we evaluated the drug exposure and distribution throughout various compartments of the murine female reproductive tract. Over a 24-h time course, <b>VU6032735</b> sustained high tissue exposure in the oviduct, where fertilization takes place. This work provides the framework for others interested in studying drug exposure and distribution throughout the female reproductive tract.</p>","PeriodicalId":36426,"journal":{"name":"ACS Pharmacology and Translational Science","volume":"8 9","pages":"3281–3295"},"PeriodicalIF":3.7,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acsptsci.5c00416","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036694","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Discovery of a Novel Anticoagulant Mechanism: Factor XI Activation Inhibitors","authors":"Ken Ellsworth,&nbsp;Peter Nizner,&nbsp;Anthony Ogawa*,&nbsp;Alan Hruza,&nbsp;Lan Wei,&nbsp;Dexi Yang,&nbsp;Matthew Lombardo,&nbsp;Lisa Sonatore,&nbsp;Christopher M. Barbieri,&nbsp;Alexandra Wickham,&nbsp;Chad Chamberlin,&nbsp;Elliott Nickbarg,&nbsp;Matthew Richards,&nbsp;Berengere Sauvagnat,&nbsp;Michael Ziebell,&nbsp;Thu Ho,&nbsp;Amjad Ali,&nbsp;Jacqueline Hicks,&nbsp;Payal Sheth and Jim Tata,&nbsp;","doi":"10.1021/acsptsci.5c00265","DOIUrl":"https://doi.org/10.1021/acsptsci.5c00265","url":null,"abstract":"<p >Despite advances in anticoagulant drug discovery to establish a new standard of care, represented by Factor Xa (FXa) inhibitors, additional therapies are needed to address the concomitant bleeding risk posed by these agents. Factor XI (FXI) plays a key role in the intrinsic coagulation cascade contributing to thrombin generation and fibril formation, and there is an increasing weight of evidence supporting a comparable efficacy and superior safety profile of FXI-based oral anticoagulants. A novel allosteric mechanism of action (MoA) for inhibiting FXI activation to Factor XIa (FXIa) was discovered by applying an AS-MS-based screen to the FXI zymogen. Biochemical and biophysical characterization of these structurally diverse small-molecule classes confirmed this mode of inhibition as specific to the FXI zymogen, and a high-resolution FXI-inhibitor cocomplex structure identified the binding pocket that is distal to the FXI cleavage site. These FXI Activation Inhibitors were active in translational functional assays, prolonging activated partial thromboplastin time (aPTT) in human plasma and demonstrating dose-dependent antithrombotic effects in the rabbit arteriovenous-shunt model of thrombosis. The results affirm that FXI Activation Inhibitors are a novel mechanism for the potential treatment of thrombotic disorders.</p>","PeriodicalId":36426,"journal":{"name":"ACS Pharmacology and Translational Science","volume":"8 9","pages":"3090–3100"},"PeriodicalIF":3.7,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ultrastable Insulin-Glucagon Fusion Protein Exploits an Endogenous Hepatic Switch to Mitigate Hypoglycemic Risk","authors":"Nicolas Varas,&nbsp;Mark A. Jarosinski,&nbsp;Yen-Shan Chen,&nbsp;Chun-Lun Ni,&nbsp;Rachel A. Grabowski,&nbsp;Ningwen Tai,&nbsp;Raimund I. Herzog,&nbsp;Faramarz Ismail-Beigi,&nbsp;Yanwu Yang,&nbsp;Alan D. Cherrington and Michael A. Weiss*,&nbsp;","doi":"10.1021/acsptsci.5c00362","DOIUrl":"https://doi.org/10.1021/acsptsci.5c00362","url":null,"abstract":"<p >The risk of hypoglycemia and its serious medical sequelae restrict insulin replacement therapy for diabetes mellitus. Such adverse clinical impact has motivated development of diverse glucose-responsive technologies, including algorithm-controlled insulin pumps linked to continuous glucose monitors (“closed-loop systems”) and glucose-sensing (“smart”) insulins. These technologies seek to optimize glycemic control while minimizing hypoglycemic risk. Here, we describe an alternative approach that exploits an endogenous glucose-dependent switch in hepatic physiology: preferential insulin signaling (under hyperglycemic conditions) <i>versus</i> preferential counter-regulatory glucagon signaling (during hypoglycemia). Motivated by prior reports of glucagon-insulin coinfusion, we designed and tested an ultrastable glucagon-insulin fusion protein whose relative hormonal activities were calibrated by respective modifications; physical stability was concurrently augmented to facilitate formulation, enhance shelf life and expand access. An N-terminal glucagon moiety was stabilized by an α-helix-compatible Lys13-Glu17 lactam bridge; a C-terminal insulin moiety was stabilized as a single chain with foreshortened C domain. Studies <i>in vitro</i> demonstrated (a) resistance to fibrillation on prolonged agitation at 37 °C and (b) dual hormonal signaling with balanced activity and cross-talk. Glucodynamic responses were monitored in rats relative to control fusion proteins lacking one or the other hormonal activity, and continuous intravenous infusion emulated basal subcutaneous therapy. Whereas efficacy in mitigating hyperglycemia was unaffected by the glucagon moiety, the fusion protein enhanced endogenous glucose production under hypoglycemic conditions. These findings provide proof of principle toward a basal glucose-responsive insulin biotechnology of striking simplicity. The fusion protein’s augmented stability promises to circumvent the costly cold chain presently constraining global insulin access.</p>","PeriodicalId":36426,"journal":{"name":"ACS Pharmacology and Translational Science","volume":"8 9","pages":"3240–3258"},"PeriodicalIF":3.7,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036688","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}