The Journal Of Pharmacology And Experimental Therapeutics[JOURNAL]
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200 papers
Machine learning-driven discovery of potent isocitrate dehydrogenase 1 mutant inhibitors from ultralarge ligand libraries for targeting malignant glioma.
Zaka M, Asaad F, Özkanca Ş … +6 more , Çelebi A, Zhou R, Kim H, Kılıç T, Avşar T, Durdağı S
J Pharmacol Exp Ther · 2026 Apr · PMID 41880802 · Publisher ↗
Glioblastoma (GBM) represents one of the most lethal and therapy-resistant forms of brain tumors, characterized by high heterogeneity, metabolic reprogramming, and recurrence. In the current study, we aimed to identify n...
Glioblastoma (GBM) represents one of the most lethal and therapy-resistant forms of brain tumors, characterized by high heterogeneity, metabolic reprogramming, and recurrence. In the current study, we aimed to identify novel small molecule inhibitors targeting mutant isocitrate dehydrogenase 1 (IDH1), a crucial enzyme involved in GBM tumor metabolism. For this aim, machine learning-based quantitative structure-activity relationships modeling was combined with structure-based e-Pharmacophore screening to virtually screen ultralarge chemical libraries containing around 157 million compounds. The best hits were selected based on docking score, predicted pIC value, and molecular mechanics/generalized Born surface area binding energy calculations. Furthermore, molecular dynamics (MD) simulations were conducted to validate the selected hit compounds. In total, 36 compounds were subjected to short MD simulations (10 ns), and 16 molecules showing low binding free energies (below -90 kcal/mol) were further analyzed through long MD simulations (100 ns). Among these, 11 synthetically available hits were ordered and experimentally tested on human glioblastoma U87 and U251 cell lines. Our experimental results showed that 5 of the tested compounds (hits 1, 4, 5, 6, and 7) reduced spheroid formation by nearly 80%-90% and inhibited cell proliferation. Moreover, these hits decreased the oxygen consumption rate and extracellular acidification rate (ECAR), by up to 62% and 55%, respectively, indicating inhibition of both mitochondrial respiration and glycolysis. Furthermore, Western blot and quantitative real-time polymerase chain reaction analyses revealed downregulation of glycolytic enzymes and stemness markers. Moreover, steered MD and free energy perturbation analyses confirmed the stable interactions of these compounds at the IDH1 mutant active site. This multistage in silico-in vitro approach allowed the identification of metabolically disruptive novel mutant IDH1 inhibitors that suppress glycolysis, mitochondrial respiration, and cancer stemness in glioblastoma cells. These compounds represent promising scaffolds for the development of next-generation GBM therapeutics. SIGNIFICANCE STATEMENT: This study integrate machine learning-guided quantitative structure-activity relationships modeling with structure-based pharmacophore screening to discover small molecule inhibitors of mutant IDH1, a central mediator of metabolic reprogramming in glioblastoma. Lead compounds identified through this pipeline inhibit mutant IDH1 activity, disrupt metabolic pathways required for glioblastoma cell viability, and concomitantly reduce stem-like phenotypes in vitro, consistent with a dual mechanism of action that targets both bulk tumor cells and cancer stem-like populations.
A mu-opioid receptor positive allosteric modulator provides opioid-sparing antinociception without enhancing opioid side effects.
Kochan KE, Clements BM, Prince TD … +4 more , Demery C, White AD, Levitt ES, Traynor JR
J Pharmacol Exp Ther · 2026 Apr · PMID 41875797 · Publisher ↗
Opioids that act at the mu-opioid receptor (MOR) are the gold standard for pain management, but can induce serious unwanted effects, including addiction liability and respiratory depression. 2-(3-bromo-4-methoxyphenyl)-3...
Opioids that act at the mu-opioid receptor (MOR) are the gold standard for pain management, but can induce serious unwanted effects, including addiction liability and respiratory depression. 2-(3-bromo-4-methoxyphenyl)-3-[(4-chlorophenyl) sulfonyl]-thiazolidine is a positive allosteric modulator of MOR that increases the actions of small-molecule opioids and opioid peptides in vitro. In vivo, 2-(3-bromo-4-methoxyphenyl)-3-[(4-chlorophenyl) sulfonyl]-thiazolidine enhances the action of endogenously released opioid peptides to provide MOR-mediated antinociception, but not constipation, reward, or respiratory depression. However, the effects of positive allosteric modulators of MOR on the behavioral actions of opioid drugs such as morphine and fentanyl have not been studied. Here, we show that 2-(3-bromo-4-methoxyphenyl)-3-[(4-chlorophenyl) sulfonyl]-thiazolidine enhances opioid drug-induced antinociception in assays for acute and inflammatory pain but not the adverse effects of constipation, respiratory depression measured by blood oxygen levels and respiration rate, or reward as determined by conditioned place preference. These data support the potential of positive allosteric modulators of MOR as effective and safe opioid-sparing agents for pain management. SIGNIFICANCE STATEMENT: The undertreatment of pain and the addiction liability of opioids necessitate new strategies to improve pain management. Here, we demonstrate that the mu-opioid receptor modulator 2-(3-bromo-4-methoxyphenyl)-3-[(4-chlorophenyl) sulfonyl]-thiazolidine enhances opioid drug-mediated analgesia in mice without enhancing constipation, reward, or respiratory depression.
Contextual Influences on Naltrexone Sensitization During Daily Morphine Exposure.
Paronis CA, Bergman J
J Pharmacol Exp Ther · 2026 Apr · PMID 41863974 · Publisher ↗
Naltrexone-precipitated withdrawal in animals that receive morphine daily can include decreases in operant responding. However, naltrexone also can decrease operant responding in untreated animals, depending on prior nal...
Naltrexone-precipitated withdrawal in animals that receive morphine daily can include decreases in operant responding. However, naltrexone also can decrease operant responding in untreated animals, depending on prior naltrexone exposure and reinforcer contingencies. The present study was conducted to further evaluate changes in the behavioral effects of naltrexone consequent to morphine exposure and changes in behavioral context. Tolerance to the effects of both μ- (morphine, heroin, buprenorphine, methadone) and κ-opioid (U50,488) agonists during daily chronic morphine also was evaluated. Squirrel monkeys (n = 4) responded under a multiple schedule consisting of 4 18-minute cycles, each comprising a 10-minute timeout period followed by 3-minutes during which fixed-ratio responding was maintained by food presentation, a 2-minute timeout period, and 3-minutes during which fixed-ratio responding was maintained by stimulus-shock termination (SST). Control response rates were comparable under both schedule conditions. All drugs decreased food-maintained responding and, albeit requiring 0.5-1 log unit higher doses, morphine, heroin, methadone, and U50,488 also decreased SST-maintained responding. Daily morphine (3.2 mg/kg/day) produced tolerance to the rate-decreasing effects of μ-opioid agonists in the absence of sensitization to naltrexone's rate-decreasing effects. Doubling the daily dose of morphine and eliminating components of SST-maintained responding resulted in a 1.5-log unit leftward shift of the naltrexone dose-effect function. Full sensitization to naltrexone's ability to decrease food-maintained responding (3-log unit leftward shift) emerged after reintroducing SST-maintained performance into daily sessions. These results indicate that naltrexone's effects on operant responding during morphine maintenance can be influenced by behavioral context as well as the level of dependence. SIGNIFICANCE STATEMENT: Repeated administration of high naltrexone doses in nonopioid dependent individuals may result in behaviorally disruptive effects of low doses (naltrexone supersensitivity). Low doses of naltrexone also have disruptive effects during opioid dependence. These studies show that the expression of naltrexone effects during opioid dependence is a product of both pharmacological and behavioral factors.
Therapeutic potential of liraglutide in rheumatoid arthritis: Modulation of inflammation, apoptosis, and metabolic dysfunction in a rat model.
Nematalla HA, Sheta E, Ghareeb AZ … +3 more , Abo Saree MM, Ghoneim AI, Elharoun M
J Pharmacol Exp Ther · 2026 Apr · PMID 41863197 · Publisher ↗
Rheumatoid arthritis (RA) is a chronic autoimmune disorder marked by joint inflammation and systemic symptoms. This study evaluates the efficacy of liraglutide (LIRA), a glucagon-like peptide-1 receptor agonist, in RA ma...
Rheumatoid arthritis (RA) is a chronic autoimmune disorder marked by joint inflammation and systemic symptoms. This study evaluates the efficacy of liraglutide (LIRA), a glucagon-like peptide-1 receptor agonist, in RA management, particularly in conjunction with methotrexate (MTX), a standard RA therapy on complete Freund's adjuvant (CFA)-induced arthritis. Rats were injected with 0.12 mL of CFA (10 mg/1 mL) intradermally on day 1. Rats were divided into 6 groups, Normal group, Model group, MTX group (methotrexate 1 mg/kg/wk/i.p.), LIRA protection group (liraglutide 75 μg/kg/day/i.p. from day 1 to day 56), LIRA group (liraglutide 75 μg/kg/day/i.p. from day 15 to day 56), LIRA + MTX group (liraglutide 75 μg/kg/day/i.p. + methotrexate 1 mg/kg/wk/i.p. from day 15 to day 56). The arthritic rats developed significant joint destruction accompanied by alterations in metabolic parameters, elevated inflammatory cytokines, and enhanced apoptosis and autophagy. Liraglutide treatment and protection significantly showed metabolic hexokinase 2-succinate-hypoxia-inducible factor 1α axis modulation, inflammasome NOD-like receptor family, pyrin domain containing 3 suppression, apoptosis and autophagy flux normalization and joint pathology improvement. Liraglutide produced more pronounced effects when administered in combination with methotrexate. In conclusion, liraglutide demonstrated significant therapeutic and protective efficacy in a CFA-induced rat model of RA. The mechanism involves metabolic reprogramming where liraglutide downregulated the hexokinase 2-succinate-hypoxia-inducible factor 1αaxis, correcting disease-associated metabolic dysregulation. Similarly, liraglutide inhibited key proinflammatory signaling cascades, specifically the nuclear factor κB/NOD-like receptor family, pyrin domain containing 3/interleukin-1β and tumor necrosis factor-α/P38 mitogen-activated protein kinase pathways. SIGNIFICANCE STATEMENT: Rheumatoid arthritis is a chronic immuno-inflammatory disorder causing joint damage. Liraglutide presents opportunities for repurposing metabolic agents in the treatment of autoimmune illnesses. Liraglutide modulates metabolic dysfunction, normalizes autophagy markers, inflammatory pathways, and lower apoptotic signals in complete Freund's adjuvant-induced arthritis in rats.
Comparison of naloxone, naltrexone, nalmefene, and methocinnamox for preventing and reversing the discriminative stimulus effects of buprenorphine and carfentanil in rats discriminating fentanyl from saline.
Hiranita T, Grisham AK, Mijares AE … +3 more , Alvarez MA, Patel MN, France CP
J Pharmacol Exp Ther · 2026 Mar · PMID 41849982 · Publisher ↗
Naloxone and nalmefene are μ opioid receptor (MOR) antagonist medications for reversing opioid overdose and naltrexone is a MOR antagonist medication for treating opioid use disorder (preventing relapse). MOR agonist med...
Naloxone and nalmefene are μ opioid receptor (MOR) antagonist medications for reversing opioid overdose and naltrexone is a MOR antagonist medication for treating opioid use disorder (preventing relapse). MOR agonist medications for treating opioid use disorder include methadone and buprenorphine. The magnitude of effect of buprenorphine can be less than the effect of higher efficacy MOR agonists with some effects of buprenorphine (eg, ventilatory depression) resistant to reversal by naloxone, compared with reversal of the effects of other MOR agonists. Drug discrimination was used to compare the ability of naloxone, naltrexone, nalmefene, and methocinnamox (MCAM) to prevent and reverse the effects of buprenorphine and the ultrapotent fentanyl analog carfentanil in male and female rats discriminating fentanyl from saline. Naloxone, naltrexone, nalmefene, and MCAM (0.01-0.1 mg/kg) were equipotent at preventing the discriminative stimulus effects of 0.01 mg/kg buprenorphine and 0.0001 mg/kg carfentanil. Compared with their potencies to prevent discriminative stimulus effects, naloxone, naltrexone, and nalmefene were ≥92-fold less potent in reversing the discriminative stimulus effects of buprenorphine and ≥31-fold less potent in reversing the discriminative stimulus effects of carfentanil. In contrast, MCAM was equipotent in preventing and reversing the discriminative stimulus effects of buprenorphine and carfentanil. There was no significant difference in the onset of action of naloxone, naltrexone, nalmefene, or MCAM. The greater potency of MCAM, compared with naloxone, naltrexone, and nalmefene for reversing the effects of buprenorphine and carfentanil might translate to MCAM reversing adverse effects of a wide range of MOR agonists in patients. SIGNIFICANCE STATEMENT: Buprenorphine, a μ opioid receptor (MOR) agonist used to treat opioid use disorder, can produce adverse effects that are reportedly more difficult to reverse with naloxone, compared with reversal of the effects of other MOR agonists. This study provides evidence for the potential utility of the MOR antagonist methocinnamox to reverse the adverse effects of buprenorphine by demonstrating the relatively greater potency of methocinnamox, compared with currently available MOR antagonists, to reverse the discriminative stimulus effects of buprenorphine and carfentanil.
Pharmacokinetic and pharmacodynamic properties of cannabigerol in male mice.
Mabou Tagne A, Ahmed F, Tran A … +8 more , Galvani F, Debbaneh L, Perranoski ER, Sarlah D, Das A, Pabon E, Cooper Z, Piomelli D
J Pharmacol Exp Ther · 2026 Apr · PMID 41833244 · Full text
Cannabigerol (CBG) is a nonintoxicating phytocannabinoid gaining popularity as a self-medication for anxiety and other conditions; however, its pharmacological properties remain poorly defined. Here, we report the develo...
Cannabigerol (CBG) is a nonintoxicating phytocannabinoid gaining popularity as a self-medication for anxiety and other conditions; however, its pharmacological properties remain poorly defined. Here, we report the development of a rapid and sensitive liquid chromatography-tandem mass spectrometry method for quantifying CBG and its primary oxidative metabolite, cyclo-CBG. This platform enabled the characterization of CBG's pharmacokinetic and biotransformation profile after intraperitoneal administration (10 mg/kg) in male mice. CBG exhibited rapid systemic distribution and clearance, with relatively low brain penetration (brain-to-plasma ratio = 0.26). In contrast, cyclo-CBG accumulated in brain tissue to a surprising extent (brain-to-plasma ratio = 7.1), suggesting local formation and a potentially important role in mediating central effects. Despite prior reports of anxiolytic effects, we found that CBG administered at its peak brain concentration produced anxiogenic-like effects in mice, as assessed using the elevated plus maze. This response was not affected by the CB cannabinoid receptor inverse agonist, rimonabant (3 mg/kg, i.p.), indicating a mechanism independent of CB signaling. As interest in CBG continues to rise, the analytical and pharmacokinetic framework presented here provides a valuable foundation for advancing preclinical and clinical investigations into its efficacy, safety, and mechanism of action. SIGNIFICANCE STATEMENT: Application of a new liquid chromatography-tandem mass spectrometry method to quantify cannabigerol reveals key pharmacokinetic properties of this phytocannabinoid in mice, including unexpectedly high brain accumulation of its metabolite cyclo-cannabigerol, which was accompanied by anxiogenic-like effects. The results offer valuable tools for advancing preclinical and clinical investigations into cannabigerol pharmacology.
Positron emission tomography-based radiopharmaceuticals for imaging the expression and function of multidrug resistance P-glycoprotein.
Muralidhar D, Kumar P
J Pharmacol Exp Ther · 2026 Apr · PMID 41831271 · Publisher ↗
Multidrug resistance is a major reason for drug resistance in patients undergoing chemotherapy or other drug therapy. The overexpression of efflux proteins, such as multidrug resistance protein or P-glycoprotein (P-gp),...
Multidrug resistance is a major reason for drug resistance in patients undergoing chemotherapy or other drug therapy. The overexpression of efflux proteins, such as multidrug resistance protein or P-glycoprotein (P-gp), has been recognized as a major cause of the drugs' efflux from the brain. P-gp expression is not only responsible for drug resistance, but underactivity leads to the accumulation of amyloid proteins and may become one of the reasons for Alzheimer disease. Hence, measuring the activity of the efflux proteins can indicate whether the candidate is suitable for chemotherapy. Therefore, several radiotracers have been developed to image P-gp activity. Positron emission tomography imaging can assess P-gp function and, therefore, plays a crucial role in informing clinicians' decisions to adjust treatment. This review article discusses advancements in radiopharmaceuticals for imaging P-gp function and expression, particularly at the blood-brain barrier. It highlights the significance of various radiolabeled tracers, including verapamil, metoclopramide, and MC225, in assessing P-gp-mediated drug delivery to the brain and its role in various neurodisorders. This article discusses the outcomes of various radiopharmaceuticals used in imaging P-gp expression and function. SIGNIFICANCE STATEMENT: This review article highlights the evolution of radiopharmaceuticals for imaging P-gp in various disorders, including drug resistance, Alzheimer disease, and epilepsy. P-gp imaging can play a crucial role in drug development and aid in identifying tumors that are responsive or resistant to chemotherapy. It may help clinicians decide whether a patient is suitable for chemotherapy. Positron emission tomography imaging can provide information on P-gp activity, which can be assessed before and during chemotherapy.
Targeting cholangiocyte sphingosine-1-phosphate (S1P) receptor 1 signaling alleviates cholestatic liver injury: Mechanistic insight into S1P/phosphorylated signal transducer and activator of transcription 3 axis.
Yuan Z, Wang J, Zhang H … +8 more , Miao Y, Chai Y, Li A, Tang Q, Chen Q, Zhang L, Yu Q, Jiang Z
J Pharmacol Exp Ther · 2026 Apr · PMID 41818851 · Publisher ↗
Effective first-line treatments for cholestasis are limited, leading to poor outcomes and liver transplantation after ursodeoxycholic acid/obeticholic acid intolerance. We investigated the role of cholangiocyte sphingosi...
Effective first-line treatments for cholestasis are limited, leading to poor outcomes and liver transplantation after ursodeoxycholic acid/obeticholic acid intolerance. We investigated the role of cholangiocyte sphingosine-1-phosphate receptor 1 (S1PR1) in cholestasis pathogenesis to identify new therapeutic targets. We generated cholangiocyte-specific S1pr1 knockout mice (S1pr1). Cholestasis models included bile duct ligation (BDL) (14 days) and 0.5% cholic acid (CA) diet (4 months). The level of sphingosine-1-phosphate (S1P) and its receptor, especially S1PR1 in cholangiocytes were significantly increased in both BDL or 0.5% CA diet models. Sphingosine kinase 1-derived sphingosine-1-phosphate from hepatic stellate cells/endothelial cells activated cholangiocyte S1PR1, promoting signal transducer and activator of transcription 3 phosphorylation and releasing interleukin-6/C-C motif chemokine 7, which remodeled the inflammatory microenvironment and exacerbated liver injury. S1PR1 deletion significantly reduced liver injury, fibrosis, and inflammation. Likewise, treatment with a specific inhibitor of cholangiocyte S1PR1, W146, slightly improved liver injury induced by BDL. The functional effect of S1PR1 in cholangiocyte was further strengthened by our design of the nanoparticle-delivered W146. This demonstrated that cholangiocytes-specific deletion of S1PR1 can alleviate liver fibrosis and injury caused by biliary obstruction or chronic cholestasis, which helps to develop S1PR1 as a target for the treatment of liver fibrosis and cholestasis. SIGNIFICANCE STATEMENT: This study identifies elevated sphingosine-1-phosphate as a potential cholestasis biomarker. High sphingosine-1-phosphate binds sphingosine-1-phosphate receptor 1, activating signal transducer and activator of transcription 3 in cholangiocytes and creating proinflammatory microenvironment. Cholangiocyte-specific sphingosine-1-phosphate receptor 1 inhibition via nanocrystal agents alleviates cholestatic liver injury.
Laxative use and acute kidney injury risk: Analysis of a Japanese hospital-based database.
Mitsuboshi S, Tsuchiya M, Kizaki H … +2 more , Hori S, Imai S
J Pharmacol Exp Ther · 2026 Mar · PMID 41806409 · Publisher ↗
Although a potential association between laxative use and the development of acute kidney injury (AKI) has been proposed, it remains inadequately investigated. Therefore, the objective of this study was to elucidate the...
Although a potential association between laxative use and the development of acute kidney injury (AKI) has been proposed, it remains inadequately investigated. Therefore, the objective of this study was to elucidate the association between the use of each laxative and the risk of developing AKI. Data were obtained from the JMDC hospital-based administrative claims database, covering the period from April 2014 to August 2022. Laxatives were defined as lubiprostone and linaclotide (secretory drugs), senna (a stimulant), and magnesium oxide (a nonabsorbed substance). AKI was defined in accordance with the recommendations of the Kidney Disease: Improving Global Outcomes working group. After screening, 159,630 patients were eligible for inclusion. AKI occurred in 693 patients (94/1000 patient-years) in the magnesium oxide group, 67 (153/1000 patient-years) in the lubiprostone group, 194 (109/1000 patient-years) in the senna group, and 22 (161/1000 patient-years) in the linaclotide group. Cox regression analysis revealed that the use of lubiprostone (adjusted hazard ratio, 0.96; 95% CI, 0.74-1.24), senna (adjusted hazard ratio, 1.08; 95% CI, 0.91-1.27), and linaclotide (adjusted hazard ratio, 1.42; 95% CI, 0.92-2.17) did not decrease the risk of AKI compared with magnesium oxide. The findings suggest that there is no difference in AKI risk among the various laxatives studied. SIGNIFICANCE STATEMENT: The results indicate that there were no differences in the risk of acute kidney injury among lubiprostone, linaclotide, senna, and magnesium oxide, suggesting that the use of these laxatives might not impact the risk of acute kidney injury.
Lomitapide reduces viability and clonogenicity in hepatocellular carcinoma cells but enhances xenograft growth: The importance of the tumor microenvironment.
Comanzo CG, Buatti Fagalde F, Vera MC … +9 more , Oviedo Bustos L, Palma NF, Capiglioni AM, Hesse L, Pisani GB, Ferretti AC, Ceballos MP, Alvarez ML, Quiroga AD
J Pharmacol Exp Ther · 2026 Apr · PMID 41795398 · Publisher ↗
Lomitapide, a microsomal triglyceride transfer protein inhibitor approved for the treatment of homozygous familial hypercholesterolemia, has recently attracted interest as a potential anticancer agent because of its effe...
Lomitapide, a microsomal triglyceride transfer protein inhibitor approved for the treatment of homozygous familial hypercholesterolemia, has recently attracted interest as a potential anticancer agent because of its effects on lipid metabolism. Given the central role of lipid handling in hepatocellular carcinoma (HCC), we investigated the impact of lomitapide-mediated microsomal triglyceride transfer protein inhibition using complementary in vitro and in vivo models. Lomitapide induced intracellular lipid accumulation and reduced cell viability and clonogenicity in human HCC cell lines (Huh7 and HepG2) in a dose-dependent manner, without affecting cell migration. However, in a subcutaneous xenograft model, lomitapide treatment paradoxically promoted tumor growth, increasing tumor volume, weight, and proliferative markers, whereas apoptosis-related proteins remained unchanged. Tumors from lomitapide-treated mice exhibited enhanced extracellular signal-regulated kinase (ERK) signaling and increased lipid accumulation, alongside reduced systemic lipoprotein levels. To reconcile these opposing effects, we examined the contribution of the tumor microenvironment. Coculture experiments revealed reduced sensitivity of HCC cells to lomitapide in the presence of nonparenchymal cells. Conditioned media studies identified hepatic stellate cells as key mediators of this resistance, associated with increased secretion of interleukin 8 and vascular endothelial growth factor after lomitapide exposure. These factors are known activators of proliferative signaling pathways in HCC. Collectively, our findings demonstrate that lomitapide exerts direct antiproliferative effects on HCC cells under simplified conditions, but promotes tumor growth in vivo through microenvironment-dependent mechanisms. This study highlights the critical influence of stromal-tumor interactions on therapeutic outcomes and urges caution in repurposing lipid-modulating drugs for cancer treatment without accounting for tissue context. SIGNIFICANCE STATEMENT: This study demonstrates that inhibition of microsomal triglyceride transfer protein exerts opposite effects on hepatocellular carcinoma depending on biological context, suppressing tumor cell growth in vitro while promoting tumor expansion in vivo. These findings reveal a decisive role for the tumor microenvironment, particularly stromal cell-derived protumorigenic signals, in shaping therapeutic responses to lipid-modulating drugs.
Adiponectin deficiency and high fat and sucrose diets impair pancreatic islet adaptations to pregnancy and contribute to gestational hyperglycemia.
Moyce Gruber BL, Fonseca MA, Trivedi K … +2 more , Doucette CA, Dolinsky VW
J Pharmacol Exp Ther · 2026 Mar · PMID 41793898 · Full text
Gestational diabetes mellitus is characterized by hyperglycemia and impaired glucose tolerance with first onset in pregnancy. Clinical research shows an association between low circulating adiponectin levels and gestatio...
Gestational diabetes mellitus is characterized by hyperglycemia and impaired glucose tolerance with first onset in pregnancy. Clinical research shows an association between low circulating adiponectin levels and gestational diabetes mellitus diagnosis. Adiponectin is a fat-derived cytokine which is postulated to improve insulin secretion and survival of pancreatic β-cells; however, the role of adiponectin in the regulation of late pregnancy glucose homeostasis requires investigation. We investigated metabolic, morphological, and functional adaptations of pancreatic islets to pregnancy in adiponectin knockout and wild-type female mice and the interaction with low fat (LF) or high fat and sucrose (HFS) diets. Pregnant wild-type mice fed a HFS diet exhibited elevated fasting glycemia and glucose intolerance, accompanied by impaired insulinogenic index at gestational day 18.5 compared to LF-fed controls, and interestingly both LF and HFS-fed adiponectin knockout mice exhibited similar impairments. Pregnancy-induced pancreatic β-cell expansion was largely preserved during late pregnancy, though the HFS diet attenuated β-cell mass at gestational day 18.5 and adiponectin deficiency triggered a similar level of impairment. Glucose stimulated insulin secretion was significantly attenuated in islets isolated from pregnant wild-type mice fed HFS diet and both LF and HFS-fed pregnant adiponectin knockout mice showed a similar level of impaired despite similar basal secretion and insulin content. Gene expression analysis of islets from pregnant mice revealed reduced Nkx6.1 and downregulation of metabolism genes (Ppargc1a, Cpt1a) and Adipor1, without changes in insulin synthesis/secretion genes or mitochondrial electron transport chain components in both the HFS-fed wild-type and both LF- and HFS-fed adiponectin knockout mice compared to wild-type LF-fed controls. These findings demonstrate that HFS feeding and adiponectin deficiency trigger a similar level of impaired insulin secretion during pregnancy that contributes to hyperglycemia, which suggests increasing adiponectin levels during pregnancy has potential therapeutic benefit. SIGNIFICANCE STATEMENT: We show that in late pregnancy, the absence of adiponectin elicits a similar level of impaired insulin secretion as high fat/sucrose diet-induced obesity, but without a synergistic effect. This suggests interventions to enhance adiponectin could improve insulin secretion and prevent gestational diabetes mellitus.
Multiomics and experimental validation reveal theophylline's mechanism targeting IL1A/ACTB/TLR4 and identify synergistic drugs in hepatocellular carcinoma.
Gao Y, Chen W, Zarogoulidis P … +8 more , Yallapu MM, Nikolic MV, Vaishnani DK, Zheng J, Nedeljkovic N, Ye K, Guo Y, Xu Q
J Pharmacol Exp Ther · 2026 Mar · PMID 41763175 · Publisher ↗
This study aimed to investigate the mechanism by which theophylline influences hepatocellular carcinoma (HCC) through the regulation of core targets and to identify its potential synergistic drugs. Integrated network pha...
This study aimed to investigate the mechanism by which theophylline influences hepatocellular carcinoma (HCC) through the regulation of core targets and to identify its potential synergistic drugs. Integrated network pharmacology, multiomics data (transcriptomics, single-cell, and spatial transcriptomics), and multiple machine learning algorithms (a total of 113 diagnostic models combined with SHapley Additive exPlanations, least absolute shrinkage and selection operator, and artificial neural network analyses) were used to screen core targets linking theophylline, gut microbiota, and HCC. Enrichment analyses (Disease Ontology, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes) were conducted to elucidate their biological functions. Molecular docking and molecular dynamics simulations were used to evaluate the binding patterns and stability between theophylline (and potential synergistic drugs) and the core targets. Databases including The University of ALabama at Birmingham CANcer data analysis Portal, Gene Expression Profiling Interactive Analysis, Comparative Toxicogenomics Database, and gutMGene were used to analyze the clinical relevance of core targets, their regulatory roles in the immune microenvironment, and their connections within the "theophylline-target-HCC chemical-gut microbiota" network. Finally, in vitro cell experiments (proliferation, migration, invasion, quantitative reverse transcription polymerase chain reaction, and Western blot) and in vivo xenograft models were used to validate the findings. Seventeen shared targets were screened, and 3 core targets, interleukin 1α, actin β, and toll like receptor 4, were further identified. Molecular docking and dynamics simulations demonstrated that theophylline could stably bind to these core targets. Multiple drugs including chlorogenic acid (PubChem CID: 1794427), losartan (PubChem CID: 3961), and estrone sulfate (PubChem CID: 3001028), were found to potentially exhibit synergistic effects with theophylline. The expression of core targets was significantly associated with clinical stage, prognosis, immune cell infiltration (eg, monocytes and macrophages), and immune checkpoints (including programmed death-ligand 1 and cytotoxic T-lymphocyte associated protein 4) in patients with HCC. Single-cell and spatial transcriptomic analyses revealed heterogeneous expression of core targets within the tumor microenvironment. Experimental validation confirmed that theophylline significantly inhibited HCC cell proliferation, migration, and invasion, downregulated core target expression, and effectively suppressed tumor growth in vivo. Theophylline may exert anti-HCC effects by regulating the core targets interleukin 1α, actin β, and toll like receptor 4, thereby influencing the tumor immune microenvironment, malignant biological behaviors of cells, and the gut microbiota-liver axis. This study provides a theoretical foundation for theophylline as a potential therapeutic or adjuvant agent for HCC and suggests directions for its combined application with specific drugs. SIGNIFICANCE STATEMENT: The research uncovers a new target and mechanism for the classic drug theophylline in treating hepatocellular carcinoma. This finding provides a key scientific basis for repurposing the drug. Furthermore, a drug combination discovered through multiomics analysis and laboratory tests offers a direct and practical new path for developing new combination therapies for hepatocellular carcinoma in the clinic.
Downregulation of osteoclast differentiation and activation by the soluble epoxide hydrolase inhibition.
Martins G, Oliveira D, Rivas CA … +5 more , Clemente-Napimoga JT, Hammock BD, Van Dyke TE, Napimoga MH, Abdalla HB
J Pharmacol Exp Ther · 2026 Mar · PMID 41763174 · Full text
This study aimed to investigate the effects of soluble epoxide hydrolase (sEH) inhibition on osteoclast differentiation and activity in vitro and in vivo, as well as to elucidate the signaling pathways associated with os...
This study aimed to investigate the effects of soluble epoxide hydrolase (sEH) inhibition on osteoclast differentiation and activity in vitro and in vivo, as well as to elucidate the signaling pathways associated with osteoclastogenesis. Primary murine bone marrow monocytes were stimulated with macrophage colony-stimulating factor and receptor activator of nuclear factor kappa B ligand to induce osteoclastogenesis and treated with the sEH inhibitor 1-(1-propanoylpiperidin-4-yl)-3-[4-(trifluoromethoxy)phenyl]urea (TPPU) (0.1-10 μM). Tartrate-resistant acid phosphatase staining, gene expression analyses, and immunofluorescence were used to evaluate osteoclast formation, transcriptional regulation, and cell fusion. A murine model of ligature-induced periodontitis was used to assess in vivo effects of sEH inhibition (TPPU 10 mg/kg). Alveolar bone loss was quantified by histomorphometry, and gingival gene expression was analyzed. In vitro, sEH inhibition significantly reduced tartrate-resistant acid phosphatase-positive multinucleated osteoclast formation, downregulated the expression of key transcription factors and osteoclast activity-related genes. Immunofluorescence analysis revealed attenuation of mitogen-activated protein kinase signaling and reduced dendritic cell-specific transmembrane protein expression, indicating impaired cell fusion. In vivo, TPPU treatment preserved alveolar bone structure, reduced osteoclast-like cell numbers, and decreased the expression of osteoclastic markers in gingival tissues during experimental periodontitis. sEH acts as a crucial regulator of osteoclast differentiation and function. Pharmacological inhibition of sEH suppresses osteoclastogenesis and protects against inflammatory bone loss. Therefore, targeting sEH may represent a novel therapeutic approach to modulate osteoclast activity and prevent bone destruction in periodontitis and other bone-resorptive diseases. SIGNIFICANCE STATEMENT: This study provides direct evidence that soluble epoxide hydrolase inhibition modulates osteoclast differentiation and fusion, contributing to reduced inflammatory bone loss. By demonstrating effects on osteoclast-intrinsic pathways while also influencing the inflammatory microenvironment, our findings support soluble epoxide hydrolase as a pharmacological target for chronic inflammatory bone-resorptive diseases.
Discovery of a pyridine-piperazine-based small molecule that enhances the activity of peptidase neurolysin.
ElSayed ElZorkany H, Kandil H, Jayaraman S … +16 more , Aly A, Esfahani SH, Patel D, Dannecker D, Maciag M, Paul A, Lowran K, Kumari S, Bose S, Ostrov DA, Wu CG, Dickson A, Abbruscato TJ, Trippier PC, Orlando BJ, Karamyan VT
J Pharmacol Exp Ther · 2026 Mar · PMID 41759240 · Full text
Neurolysin (Nln) is a peptidase recognized for its cerebroprotective function in acute ischemic stroke. This study aimed to identify small molecule activators of Nln as research tools to further explore the role of this...
Neurolysin (Nln) is a peptidase recognized for its cerebroprotective function in acute ischemic stroke. This study aimed to identify small molecule activators of Nln as research tools to further explore the role of this enzyme in stroke and other neurological disorders. Building on our previous computational screen of ∼140,000 compounds from the National Cancer Institute Developmental Therapeutics Program database, we extended experimental testing to the top 100 candidates using an Nln enzymatic assay. A pyridine-piperazine derivative (Py-Pip) was identified as a hit molecule and was characterized in detail. Py-Pip concentration-dependently enhanced the hydrolysis of both synthetic and natural substrates (neurotensin, angiotensin I, and bradykinin) by rat Nln, and displayed comparable activating effects on human and mouse orthologs. Importantly, Py-Pip exhibited a favorable selectivity profile, showing no potentiation of homologous metallopeptidases or unrelated enzymes. Kinetic analysis revealed that Py-Pip increases the catalytic efficiency (V/K) of Nln via a nonessential activation mechanism, whereas competition assays with inhibitor dynorphin A(1-13) confirmed that Py-Pip acts at a distinct, nonoverlapping site. Direct binding was further validated by orthogonal biophysical techniques, including differential scanning fluorimetry, microscale thermophoresis, and biolayer interferometry, whereas circular dichroism spectroscopy indicated activator-induced secondary structural changes. These findings validate that Nln activity can be enhanced by small molecules and establish Py-Pip as a novel, nonpeptide scaffold for developing potent, "drug-like" activators to investigate Nln biology and therapeutic potential. SIGNIFICANCE STATEMENT: This study reports the discovery of a novel nonpeptide small molecule that selectively enhances the activity of neurolysin (Nln), a peptidase implicated in cerebroprotection. Unlike previous peptide-based activators, this molecule provides a stable, "drug-like" scaffold and a structural foundation for the development of potent Nln activators to probe Nln biology and therapeutic potential in ischemic stroke.
Pharmacogenetic influence on bupropion bioactivation and clinical outcomes in major depressive disorder.
Kharasch ED, Lenze EJ
J Pharmacol Exp Ther · 2026 Mar · PMID 41759239 · Publisher ↗
This investigation addressed influences of CYP2B6 and CYP2C19 genetic polymorphisms on steady-state bupropion disposition, antidepressant outcomes and side effects, and evaluated influences of pharmacokinetic variability...
This investigation addressed influences of CYP2B6 and CYP2C19 genetic polymorphisms on steady-state bupropion disposition, antidepressant outcomes and side effects, and evaluated influences of pharmacokinetic variability on pharmacogenetic results. This was a preplanned secondary analysis of a prospective, 24-week, randomized, double-blind, crossover trial which compared pharmacokinetics and antidepressant effects across brand and 3 generic bupropion XL 300 mg drug products in 70 participants with major depressive disorder in remission, who were genotyped for CYP2B6 and CYP2C19 polymorphisms. We measured steady-state enantiomeric plasma and urine bupropion and primary (hydroxybupropion, a bioactivation pathway), reduced, and secondary metabolites, clinical depression (Montgomery-Asberg depression rating scale), side effects, and influence of CYP2B6 and CYP2C19 polymorphisms on bupropion disposition and drug effects across all 4 drug products (pooled analysis). Results showed that CYP2B6 polymorphisms (CYP2B6∗6 allele and the 516G>T variant alone) had a significant influence on bupropion disposition, including apparent oral clearance and enantiomers hydroxylation. Since bioactivation via bupropion hydroxylation has therapeutic importance, CYP2B6 polymorphisms could be clinically relevant. However, there were no significant differences between CYP2B6∗6 or 516G>T gene groups in either depression or side effect scores, at baseline on patients' own bupropion, or during the crossover phase of 4 bupropion products. CYP2C19 polymorphisms had a small and clinically unimportant influence on bupropion minor metabolites and secondary metabolism, and no effect on depression or side effect scores. Results show that CYP2B6 polymorphisms significantly influenced bupropion enantiomers steady-state disposition, but not long-term antidepressant or side effects. These findings may have relevance for clinical pharmacogenetic testing in depression. SIGNIFICANCE STATEMENT: This pharmacogenetic study in major depressive disorder evaluated both pharmacokinetics and clinical effects. CYP2B6 polymorphisms (CYP2B6∗6 allele, 516G>T variant) had significant influence on steady-state bupropion enantiomers apparent oral clearance and hydroxylation, but there was no association between CYP2B6∗6 or 516G>T gene groups and either depression or drug side effects. CYP2C19 polymorphisms had a small and clinically unimportant influence on bupropion minor metabolites and secondary metabolism and no effect on depression or side effect scores. These results may have implications for genetic testing in bupropion pharmacotherapy of major depressive disorder.
Moving beyond shock: Rich-lean transition procedures as a modern approach to anxiolytic screening.
Rough MI, Kratzenberg JL
J Pharmacol Exp Ther · 2026 Mar · PMID 41740199 · Full text
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Selective serotonin reuptake inhibitor fluoxetine, reduces solid tumor burden and metastasis through activation of host antitumor immune response and modulation of tumor microenvironment.
Mahanti K, Saha J, Mondal P … +5 more , Sarkar D, Pramanik A, Das D, Mahato M, Bhattacharyya S
J Pharmacol Exp Ther · 2026 Mar · PMID 41734401 · Publisher ↗
The tumor microenvironment (TME) plays a pivotal role in tumorigenesis, metastasis, and resistance to therapy. Although immunotherapeutic approaches such as CAR-T cells, immune checkpoint inhibitors, and tumor vaccines h...
The tumor microenvironment (TME) plays a pivotal role in tumorigenesis, metastasis, and resistance to therapy. Although immunotherapeutic approaches such as CAR-T cells, immune checkpoint inhibitors, and tumor vaccines have demonstrated promising therapeutic effectiveness, tumor heterogeneity and immunosuppression continue to limit their efficacy in solid tumors, therefore, development of effective low-cost TME targeted therapy is necessary. Recent studies indicate, selective serotonin reuptake inhibitors display immune modulatory effects. Fluoxetine is an US Food and Drug Administration-approved selective serotonin reuptake inhibitor widely used in treatments of depression in patients with cancer, its effects toward immune responses in TME and effectiveness in management of solid tumor has not been explored previously. In this study, we investigated the immunoregulatory and antitumor effects of fluoxetine on solid tumor using syngeneic orthotropic B16-F10 melanoma and 4T1 breast carcinoma models in mice. Fluoxetine oral application significantly decreased the primary tumor burden and development of metastatic nodules. Histopathology, flowcytometry and confocal imaging revealed that fluoxetine significantly activates host antitumor immune system through increased cytotoxic interferon gamma secreting T cells and M1-macrophages accumulation at both primary and metastatic sites that results in elevated activated caspase-8 in cancer cells leading to immune mediated tumor cell death. Fluoxetine treatment led to decrease in immunosuppressive cells populations such as myeloid derived suppressor cells, M2-macrophages, regulatory T cells, while also reducing the level of cancer stemness markers and proteins associated with epithelia to mesenchymal transition and metastasis. Collectively, our study indicates the multifaceted role of fluoxetine in modulating the solid TME and highlight its potential in designing of novel anticancer treatment regimen. SIGNIFICANCE STATEMENT: Fluoxetine, an US Food and Drug Administration-approved selective serotonin reuptake inhibitor reduces solid tumor (breast and melanoma) load through immune mediated tumor cell death and restore antitumor immunity. It remodels tumor microenvironment, activates T cells, reduces cancer stemness and metastasis via regulation of epithelial mesenchymal transition factors, hypoxia Inducible factor 1α, and β-catenin.
Flavorants enhance nicotine vapor self-administration in diet-induced obese female mice.
Tetteh-Quarshie S, Mensah EN, Booker DM … +1 more , Henderson BJ
J Pharmacol Exp Ther · 2026 Mar · PMID 41719691 · Publisher ↗
Nicotine dependence and obesity are global health issues. Emerging studies suggest that obese individuals are more likely to develop nicotine dependence. Despite this knowledge, there have been few investigations into wh...
Nicotine dependence and obesity are global health issues. Emerging studies suggest that obese individuals are more likely to develop nicotine dependence. Despite this knowledge, there have been few investigations into whether nicotine dose and/or chemical flavorants commonly found in nicotine-containing products modulate nicotine dependence in obese models. Therefore, our objective was to examine whether low (6 mg/mL) or high (60 mg/mL) dose nicotine, with or without flavor impact vaping-related behaviors in a diet-induced obesity (DIO) mouse model. Briefly, adult male and female DIO mice were trained to self-administer 6 or 60 mg/mL nicotine, with or without menthol and green apple (GA) flavors using an e-Vape self-administration assay. Overall, DIO female mice assigned to flavored e-liquids self-administered more nicotine than their male counterparts. Additionally, DIO female mice assigned to 6 mg/mL nicotine + menthol exhibited higher nicotine reinforcement-related behavior than female mice assigned 6 mg/mL nicotine (unflavored). Reinforcement-related behavior was not changed in DIO male mice assigned to e-liquids containing 60 mg/mL nicotine. However, DIO female mice exhibited menthol- and GA-induced enhancements in reinforcement-related behavior with 60 mg/mL nicotine. By combining a noncontingent vapor exposure assay with fast-scan cyclic voltammetry, we observed that menthol and GA flavorants, when combined with 6 or 60 mg/mL nicotine, increased tonic- and phasic-stimulated dopamine signaling in the nucleus accumbens core. Together, these preclinical findings provide additional evidence that nicotine plus popular chemical flavorants such as menthol and GA could alter dopamine signaling in the reward pathway and promote vaping-related behaviors in a DIO mouse model. SIGNIFICANCE STATEMENT: This study highlights the effects of flavorants on vaping-related behaviors in a diet-induced obese mouse model. Collectively, this study revealed that flavorants commonly found in vaping products alter dopamine release in the nucleus accumbens core-a phenomenon linked with nicotine addiction.
Empagliflozin mitigates doxorubicin-induced nephrotoxicity by modulating sirtuin 1, nuclear factor-κB/tumor necrosis factor-α, and cleaved caspase-3 pathways without compromising its cytotoxic efficacy.
Tawfik AG, Habib HA, Zeidan EM … +1 more , Heeba GH
J Pharmacol Exp Ther · 2026 Mar · PMID 41702351 · Publisher ↗
Doxorubicin (DOX), an anthracycline antibiotic, is a first-line chemotherapy drug for managing various tumors. Its hazards, particularly nephrotoxicity, complicate its clinical application. Empagliflozin (EMPA), an antid...
Doxorubicin (DOX), an anthracycline antibiotic, is a first-line chemotherapy drug for managing various tumors. Its hazards, particularly nephrotoxicity, complicate its clinical application. Empagliflozin (EMPA), an antidiabetic drug inhibiting sodium-glucose cotransporter 2, exhibits multiple benefits in different illnesses based on its ability to mitigate oxidative injury, inflammation, and apoptosis. Therefore, this work was conducted to evaluate EMPA's therapeutic efficacy in abrogating DOX-elicited renal impairment and to elucidate the underlying mechanisms mediating this proposed ameliorative effect. Thirty-four male Wistar rats randomly divided into control, EMPA (10 mg/kg/d, intragastric (i.g.), DOX (20 mg/kg, single intraperitoneal injection on day 10), and DOX + EMPA (10 mg/kg/d of EMPA, i.g. administration for 12 days and 20 mg/kg, single intraperitoneal injection of DOX on day 10) were involved. In vitro cytotoxicity assay was also performed using KMH2, MG63, and MCF7 cancer cell lines. Counteraction of DOX-impaired renal function by EMPA was proved by a decline in levels of serum urea, creatinine, and cystatin C levels, as well as preserved renal architecture. EMPA mitigated oxidative stress in renal tissues, as presented by a reduction in malondialdehyde and an increase in both reduced glutathione and superoxide dismutase. Moreover, EMPA alleviated DOX-induced downregulation of sirtuin 1 and upregulation of nuclear factor-κB, tumor necrosis factor-α, and cleaved caspase-3. Additionally, the cytotoxic activity of DOX in KMH2 and MG63 cancer cell lines was significantly (P < .05) promoted by EMPA. Collectively, antioxidant, anti-inflammatory, and antiapoptotic influences contributed to the preservative potential of EMPA against DOX-induced kidney toxicity. Moreover, EMPA enhanced the DOX cytotoxic impact in different cancer cell lines. SIGNIFICANCE STATEMENT: This study demonstrates that empagliflozin attenuates doxorubicin (DOX)-induced renal impairment in rats and promotes DOX's cytotoxic effects via its antioxidant and anti-inflammatory potentials. These preclinical findings highlight empagliflozin as a hopeful therapeutic tool to preserve the kidney during DOX treatment.
Preclinical characterization of pharmacokinetics, enzyme occupancy, and pharmacodynamics of soticlestat (TAK-935), a novel cholesterol 24-hydroxylase inhibitor.
Miyamoto M, Toshiya Nishi, Koike T … +15 more , Ikeda S, Sunahara E, Watanabe S, Hasegawa S, Ishii T, Kosugi Y, Yamamoto S, Fujioka Y, Igari T, Sano N, Furuta A, Amano N, Okuda T, Takahashi J, Hirabayashi H
J Pharmacol Exp Ther · 2026 Mar · PMID 41702350 · Publisher ↗
The conversion of cholesterol to 24S-hydroxycholesterol (24HC) is a brain-specific reaction catalyzed by cholesterol 24-hydroxylase (CH24H). Soticlestat, a specific inhibitor of CH24H, has the potential to reduce 24HC le...
The conversion of cholesterol to 24S-hydroxycholesterol (24HC) is a brain-specific reaction catalyzed by cholesterol 24-hydroxylase (CH24H). Soticlestat, a specific inhibitor of CH24H, has the potential to reduce 24HC levels in the brain; 24HC is a neuroactive oxysterol that modulates several receptors and ion channels involved in regulating neural excitability. Soticlestat has been investigated as an additional treatment option for seizures associated with Dravet and Lennox-Gastaut syndromes. Comprehensive preclinical evaluation of pharmacokinetics (PK), target enzyme occupancy (EO), and pharmacodynamics (PDs)-including confirmation of their interrelationships within the brain as the target tissue-is essential for elucidating the mechanism of action of soticlestat. Preclinical data on PK, EO, and PD in rodent models facilitate translational modeling, thereby informing rational human dose selection. A divergence in the temporal profiles of soticlestat concentrations between brain and plasma was observed, with prolonged brain exposure relative to plasma clearance, consistent with the compound's slow dissociation from CH24H. Herein, we characterized the PK, PD, and EO profiles of soticlestat in rodents and demonstrated the relationship between CH24H inhibition and 24HC reduction. Soticlestat demonstrated a delayed-effect model in mice, with time lags between PK and PD as well as between brain and plasma pharmacokinetics. Subsequently, the steady-state PK/PD analysis after subcutaneous infusion in CH24H wild-type [CH24H] and knockout [CH24H] mice provided comprehensive insights into the model linking the PK, EO, and PD profiles of soticlestat. SIGNIFICANCE STATEMENT: In the present study, a clear relationship between the pharmacokinetics, target enzyme occupancy, and pharmacodynamic markers of cholesterol 24-hydroxylase for soticlestat in rodents. The preclinical pharmacokinetic/enzyme occupancy/pharmacodynamic data for soticlestat provide comprehensive mechanistic and quantitative insights into clinical population models. Importantly, these data emphasize the potential of enzyme occupancy and pharmacodynamics as strategic tools for facilitating translational research in central nervous system drug development.
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