Understanding the mechanisms of drug transfer across the human placenta is critical for evaluating foetal exposure and developmental safety. Sodium-glucose co-transporter 2 inhibitors (SGLT2i), a novel class of antidiabetics, remain poorly characterized with respect to their placental transfer and potential effect on placental function. Here, we address this knowledge gap. We investigated placental transfer mechanisms of dapagliflozin (DAPA), empagliflozin (EMPA), ertugliflozin (ERTU) and canagliflozin (CANA) using term placentas and complementary in vitro assays. All SGLT2i crossed the barrier within four hours of perfusion at clinically relevant concentrations. DAPA, the smallest compound, showed the fastest transfer, whereas EMPA, the largest, was the slowest. The more lipophilic compounds, CANA and ERTU, accumulated strongly in placental tissue. Transport of all SGLT2i - especially EMPA and ERTU - increased with the addition of an ATP-synthesis inhibitor, indicating the influence of active efflux transporters. Additionally, transport was significantly enhanced under low-bovine serum albumin conditions, implicating protein binding as a key rate-limiting factor. During Transwell® permeability assays, all SGLT2i crossed the BeWo b30 cell layer that mimics early gestational cytotrophoblasts. BeWo cells exposed to SGLT2i showed no impairment in forskolin-induced differentiation, viability, or secretion of pregnancy-related hormones, but revealed increased glycolytic activity. Our findings indicate that at clinically relevant concentrations, SGLT2i cross the placenta by passive diffusion, modulated by efflux transporters and protein binding, and might alter placental glycolytic activity without impairing viability, differentiation, or hormone secretion. Their use during pregnancy should remain contraindicated, but if attempted, monitoring unbound drug concentrations is advised.

Purpose: The objective of the present study is to investigate antiproliferative effects of empagliflozin (EMPA) and to determine whether it induces autophagic cell death in lung cancer cells. Materials and Methods: A549 lung cancer cells were treated with increasing concentrations of EMPA, and cell viability was assessed by MTT assay. Autophagy and apoptosis related markers (LC3B, caspase-3, caspase-9) were examined by western blotting. Clonogenic assays were performed following treatment with EMPA alone or in combination with everolimus (RAD001) or hydroxychloroquine (HCQ). Results: EMPA significantly reduced A549 cell viability in a dose-dependent manner. The IC50 values were 54.11 µM at 24 h and 109.6 µM at 48 h. EMPA also markedly decreased clonogenic capacity by 66% and induced morphological alterations. In addition, the expression levels of LC3B and cleaved caspase-3 were increased. Our findings demonstrate that EMPA activates autophagy and triggers caspase-3 dependent apoptotic cell death. Conclusion: Empagliflozin exerts significant antiproliferative activity in lung cancer cells through the induction of autophagy and caspase-3 mediated apoptosis. Its ability to modulate autophagic and apoptotic pathways suggests that EMPA may represent a potential candidate for future combination based therapeutic strategies in lung cancer. Further mechanistic investigations and in vivo studies are warranted to validate these findings.

Simultaneous therapeutic drug monitoring of metformin (MEF) and empagliflozin (EMP) in plasma requires sustainable, white analytical chemistry (WAC) methods. Existing techniques suffer from poor greenness, electrode modification complexity, and limited plasma applicability. White/green differential pulse voltammetry (DPV) method developed using unmodified glassy carbon electrode (GCE). Protein precipitation (acetonitrile) from ethically sourced blank human plasma (BUC-IACUCPHA183A/2025). Optimized: phosphate buffer pH 7, 75 mV/s scan rate. Validated per ICH Q2(R1) across linearity, LOD/LOQ, accuracy, precision, specificity (n = 9). Linearity: MEF 20-60 μM (r = 0.997), EMP 10-70 μM (r = 0.997). LODs: 4.38 μM (MEF), 4.15 μM (EMP). Plasma recoveries: 99.4 ± 1.5% (MEF), 100.4 ± 1.4% (EMP), n = 5. Whiteness: 97/100 (RGB-12); greenness: 83/100 (MOGABI) - superior to reported methods. Validated DPV-GCE enables rapid, sustainable MEF/EMP quantification in plasma without modification/chromatography. Proof-of-concept for routine therapeutic monitoring; clinical translation limited by spiked (not patient) samples.

Chronic kidney disease (CKD) in persons with diabetes mellitus (DM) continues to be the leading cause of end-stage kidney disease (ESKD) worldwide. Despite the use of reninangiotensin system blockade, sodium-glucose transport protein 2 inhibitors (SGLT2i), glucagon-like peptide-1 receptor agonists, and nonsteroidal mineralocorticoid receptor antagonists the risk of progression to ESKD and CV events persists in this high-risk population. For this reason, new or combined strategies for treating persons with CKD and DM are in constant development. This editorial discusses the results of a recent published trial, Combination Effect of Finerenone and Empagliflozin (EMP) in Participants with Chronic Kidney Disease and Type 2 Diabetes Using a Urinary Albumin-to-Creatinine Ratio (UACR) Endpoint (CONFIDENCE) in persons with DM and CKD focused in the mechanisms involved in the beneficial renal effect in terms of albuminuria reduction[1]. Whether there is related to the synergistic effect of the combination therapy (EMP + finerenone) or merely a manifestation secondary to systolic blood pressure (SBP) reduction remains to be determined[2].The CONFIDENCE trial, a double-blind, multicenter, study involving 800 patients with type 2 DM and CKD (eGFR 30 to 90 ml/min/1.73m 2 and urinary albumin-to creatinine ratio (UACR) 100 to < 5000 mg/g) tested the effect of combining finerenone and EMP on reducing albuminuria compared with monotherapy with either empagliflozin or finerenone alone, over 180 days. All patients were on maximum tolerated angiotensin II blockade at baseline. The primary endpoint was the relative change in the log-transformed mean UACR from baseline to 180 days, a surrogate marker for kidney disease progression in previous meta-analysis [3].

Background and objectiveInflammation drives early recurrent cardiovascular risk in type 2 diabetes mellitus (T2DM) patients following acute myocardial infarction (AMI), particularly within 30–90 days post-discharge. Sodium-glucose co-transporter 2 (SGLT2) inhibitors such as empagliflozin (EMPA) provide cardiometabolic benefits, but their anti-inflammatory effects and optimal timing after AMI remain unclear. Given the prognostic role of systemic markers like the neutrophil-to-lymphocyte ratio, we investigated whether early initiation of EMPA modulates NOD-like receptor protein-3 (NLRP3) inflammasome activity and inflammatory responses in monocyte-derived macrophages (MDMs) from T2DM-AMI patients.MethodsSixty-six participants were randomised to receive EMPA either at discharge (Arm-A) or following a 90-day delay (Arm B). Clinical data and biological samples were collected over 180 days. CD14+ MDMs and plasma were obtained at days 0, 30, and 90 (EMPA vs. no EMPA), and days 90, 120, and 180 (early vs. delayed). Inflammatory and metabolic markers were assessed using RT-qPCR, luminescence-based caspase-1 and ATP assays, and targeted immunoassays.ResultsEarly EMPA administration was associated with reduced NLRP3 priming (IL1β mRNA) and activation (caspase-1 activity), potentially linked to decreased release of ATP, a danger associated molecular pattern (DAMP). In the absence of EMPA, pro-inflammatory cytokines (TNFα, IL6, MCP1) and M1 macrophage markers (e.g., CD80) either increased or remained unchanged over time. Early EMPA treatment appeared to stabilise or reduce their expression. Markers of cell senescence (p21, IL8, BCL2) were also modulated. Plasma levels of senescence-associated markers (MMP9, OPN, Serpin E1) remained largely unchanged, highlighting the importance of evaluating macrophage-specific responses.ConclusionEarly empagliflozin administration in T2DM-AMI patients was associated with modulation of NLRP3-related inflammatory and senescence pathways in patient-derived macrophages, benefits observed when cells were stimulated ex-vivo with an inflammatory stimulus. These findings provide mechanistic insight into the timing-dependent anti-inflammatory effects of EMPA and underscore its potential for immediate post-AMI use to reduce inflammation and lower residual cardiovascular risk, supporting further clinical investigation.Graphical abstractSupplementary InformationThe online version contains supplementary material available at 10.1186/s12933-025-03042-7.

Two simple, rapid, cost-effective, and environmentally friendly chromatographic methods were developed and validated for the simultaneous determination of metformin (MEF), linagliptin (LIN), and empagliflozin (EMP) in human plasma, with successful application to pharmacokinetic study. Plasma sample preparation was performed using a straightforward protein precipitation technique employing acetonitrile: methanol: trichloroacetic acid (50:49:1, by volume), which provided high extraction recovery and minimal matrix interference. The first method was based on high-performance liquid chromatography with diode array detection (HPLC-DAD) using an ODS Hypersil C18 column and isocratic elution with a mobile phase consisting of acetonitrile, methanol, and phosphate buffer (pH 3) in a ratio of (40:40:20, by volume), at a flow rate of 1.3 mL/min, with detection at 230nm. The second method employed high-performance thin-layer chromatography (HPTLC) with densitometric detection at 225nm, using silica gel 60 F254 plates and n-hexane: methanol: glacial acetic acid (6:3:1, by volume) as the developing system. Excellent linearity was achieved over concentration ranges of 85-1650 ng/mL for MEF, 50-1100 ng/mL for EMP, and 45-950 ng/mL for LIN using the HPLC method, and 500-2800, 100-800, and 50-550 ng/band, respectively, using the HPTLC method, with correlation coefficients exceeding 0.998. The lower limits of quantitation for the HPLC method were 85, 50, and 45 ng/mL for MEF, EMP, and LIN, respectively. Both methods demonstrated satisfactory accuracy, precision, recovery (> 92%), stability, and negligible matrix effects in accordance with European Medicines Agency guidelines. The validated methods were successfully applied to a pharmacokinetic study in healthy volunteers, yielding mean Cmax values of 877.5 ± 162.2 ng/mL (MEF), 576 ± 87.5 ng/mL (EMP), and 680.8 ± 7.9 ng/mL (LIN), with Tmax values of 2.42 ± 0.38, 1.5 ± 0.61, and 5.3 ± 0.52h, respectively. The obtained pharmacokinetic parameters were consistent with reported literature, confirming the reliability and clinical applicability of the proposed green bioanalytical methods.

Background: Aberrant protein glycosylation mediated by the UDP-N-acetylgalactosamine:polypeptide N-acetylgalactosaminyltransferase (GALNT) family has been implicated in the pathogenesis of metabolic syndrome (MetS), yet the roles of GALNT2 and GALNT3 remain poorly characterized. Objective: This study aimed to evaluate the modulatory effects of decaffeinated green tea and green coffee on GALNT2 and GALNT3 mRNA expression in MetS rats, compared with standard therapies, metformin and empagliflozin. Methods: Twenty-five male Sprague Dawley (SD) rats were divided into five groups randomly: (1) MetS rats (METS), (2) MetS rats treated with 300 mg/kgBW decaffeinated green tea and 200 mg/kgBW decaffeinated green coffee (TKD), (3) MetS rats treated with 500 mg/kgBW metformin (MFN), (4) MetS rats treated with 30 mg/kgBW empagliflozin (EMPA), and (5) negative control (NC). The MetS model was induced using a high-fat, high-sucrose (HFHS) diet followed by streptozotocin injection (30 mg/kgBW). After 10 weeks, the treatment groups received interventions for 9 weeks. GALNT2 and GALNT3 mRNA expression was investigated using reverse transcription polymerase chain reaction (RT-PCR).. Result: TKD and MFN tended to decrease GALNT2 and increase GALNT3 mRNA expression, although the differences were not significant statistically compared to the METS group (p&gt;0.05). EMPA decreased GALNT2 and increased GALNT3 mRNA expression significantly compared to the METS group (p&lt;0.05). Conclusion: The significant effect of EMPA on GALNT2 and GALNT3 mRNA expression suggests its therapeutic potential in targeting glycosylation pathways in MetS. Although TKD and MFN exhibited similar trends with EMPA, the lack of statistical significance suggests the need for further study. Keyword : GALNT2; GALNT3; Green Coffee; Green Tea; Metabolic Syndrome.

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.

This study aims to see the individual and combined effects of Angiotensin II type 2 (AT2) receptor agonist buloxibutid (also known as Compound 21 or C21) and sodium-glucose co-transporter-2 (SGLT-2) inhibitor empagliflozin (EMPA) on effects of hypertension (HT), which is common today, on the heart, and vascular tissue. Male rats of the Sprague Dawley were divided into 5 groups: Control (C) group, HT group, HT + C21 group, HT + EMPA group and HT + C21 + EMPA group. After the protocol was completed, hemodynamic measurements were taken and heart and aorta tissues were evaluated biochemically, histopatologically and immunohistochemically. When the mean blood pressure (BP) values were compared, the mean BP of the HT group increased significantly compared to the C group (p < 0.05). Superoxide dismutase, glutathione and glutathione peroxidase activities in the heart, glutathione and catalase activities in the descending aorta were significantly higher in all treated groups compared to the HT group (p < 0.05). In the HT + C21 + EMPA group, histopathological damage score in hematoxylin-eosin (HE) stained heart sections compared to the HT group showed a decrease in tissue damage but cell infiltration was still observed. When HE staining method was applied, it was determined that the thoracic aorta sections in group C had normal histologic structure. In the HT group, dilatation in some parts and irregularities in elastic lamellae were observed. It was observed that the treated groups were similar to group C. When considering the individual and combined effects of C21 and EMPA, positive results on heart and vascular tissue were observed by hemodynamic, biochemical and histopathological analyses.

Background/Objectives: Empagliflozin (EMPA) was repurposed for Alzheimer’s disease (AD) treatment via buccal delivery, exploiting novel nanofibers (NFs) integrating chitosan (Cs), silk fibroin (Fb), and poly(lactic acid) (PLA). Methods: EMPA-loaded Cs/Fb/PLA NFs were electrospun in different formulations to optimize the formulation parameters. The optimized formulation was then investigated for its enhanced in vivo effect. Results: Optimized nanofiber diameters ranged from 459 ± 173 to 668 ± 148 nm, possessing bead-free morphology confirmed by SEM and satisfactory mechanical properties. EMPA was successfully well-dispersed in the polymer matrix as evidenced by FTIR, XRD, and drug content. The optimized NFs displayed a hydrophilic surface (contact angle < 90°), and biphasic drug release with sustained EMPA liberation (84.98% over 24 h). In vivo, buccal EMPA-Cs/Fb/PLA NFs in an AlCl3-induced AD rat model significantly reduced brain-amyloid-β, phosphorylated tau, IL-1β, and AGER expression by 2.88-, 2.64-, 2.87-, and 2.50-fold, respectively, compared to positive controls, and improved locomotor activity (1.86-fold) and cognitive performance (T-maze) (4.17-fold). Compared to pure EMPA, the nanofiber formulation achieved further reductions in amyloid-β (1.78-fold), p-tau (1.42-fold), IL-1β (1.89-fold), and AGER (1.38-fold), with efficacy comparable to memantine. Histopathological examination revealed preservation of the hippocampal neuronal structure. Conclusions: The findings suggest EMPA-loaded Cs/Fb/PLA NFs as a promising non-invasive, sustained-release buccal delivery platform for AD therapy, offering multimodal neuroprotection through modulation of the Aβ–AGER–p-tau axis.

Arsenic trioxide (ATO), utilized in the treatment of acute promyelocytic leukemia (APL), presents significant renal toxicity that restricts its clinical usage. The potential effects of empagliflozin (EMPA) on ATO‐induced nephrotoxicity remain unexplored. This study aims to investigate whether EMPA can alleviate ATO‐induced nephrotoxicity in both animal and cellular models, as well as to further explore the underlying mechanisms. EMPA can improve renal function in mice and alleviate ATO‐induced structural damage to the kidneys. EMPA treatment effectively inhibits ATO‐induced oxidative stress and reduces apoptosis. EMPA significantly decreases the production of ROS in cultured HEK293T cells, lowers the apoptotic rate, and safeguards mitochondrial function. EMPA upregulates the SIRT1/Akt/Nrf2 pathway and addresses ATO‐induced autophagy dysfunction. These findings suggest that EMPA may ameliorate ATO‐induced renal toxicity by activating the SIRT1/Akt/Nrf2 signaling pathway, which is associated with the suppression of oxidative stress, reduction of apoptosis, and protection of mitochondrial functionality.

ABSTRACT A robust, accurate, linear, and stability‐indicating liquid chromatography (LC) method equipped with a photodiode array (PDA) detector was developed and validated for the quantitative estimation of empagliflozin (EMP), an antidiabetic agent approved for the treatment of type 2 diabetes mellitus. The method enables the determination of specified known impurities as well as degradation products present in the EMP drug substance. A systematic Quality by Design (QbD) approach was employed to evaluate the method's robustness, ensuring consistent performance under varied analytical conditions. Chromatographic separation was achieved using a Zorbax SB‐C18 column (250 × 4.6 mm, 3.5 µm) with a gradient elution program. The mobile phase consisted of Solvent A (water: acetonitrile, 50:50 v/v) and Solvent B (methanol: water, 90:10 v/v). The optimized chromatographic conditions included a flow rate of 0.7 mL/min, column temperature of 27°C, injection volume of 20 µL, and UV detection at 225 nm. Method validation was performed in accordance with ICH Q2(R2) guidelines. The method demonstrated excellent accuracy, with recovery values ranging from 95.8% to 105.4%, and linearity was observed over a wide concentration range from the limit of quantification (LOQ) to 300% of the target level, with correlation coefficients ( R 2 ) exceeding 0.998 for all identified impurities. Method precision, expressed as relative standard deviation (RSD), was consistently below 5.0%. Forced degradation studies confirmed that EMP is stable under various chemical and physical stress conditions, further supporting the stability‐indicating nature of the developed method.

IntroductionCisplatin (CP) is a chemotherapeutic agent limited by its toxic effects on the heart and kidneys, resulting in nephrotoxicity and cardiotoxicity. This study investigated the protective effects of the antidiabetic drugs linagliptin (LINA) and empagliflozin (EMPA), alone and in combination, on cisplatin-induced heart and kidney damage in non-diabetic rats.MethodsForty-nine rats were randomized into seven groups (n=7 each): control (saline; 10 mL/kg), cisplatin (CP; 10 mg/kg), empagliflozin (EMPA; 10 mg/kg), linagliptin (LINA; 3 mg/kg), and treatment groups (CP+EMPA, CP+LINA, and CP+EMPA+LINA). EMPA and LINA were administered orally for 10 days; CP was administered intraperitoneally on days 1 and 6 after treatment with EMPA, LINA, or both. Renal and cardiac injury biomarkers were measured in the blood. Hearts and kidney tissues from animals were extracted to assess reactive oxygen species (ROS) and malondialdehyde (MDA) concentrations, as well as the activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX).ResultsThe CP-treated group showed elevated renal, cardiac, and oxidative stress markers. Conversely, the activities of SOD, GPX, and CAT were reduced in the CP-treated groups compared to the control. EMPA, LINA, or combination reduced creatinine, BUN, LDH, CK-MB, troponin I, and ROS levels. LINA showed weaker antioxidant effects in the heart than in the kidney or EMPA.ConclusionOur findings suggest that LINA may have a weaker antioxidant effect in the heart than in the kidney or EMPA. Administration of the combinations (LINA+EMPA) affects MDA levels in CP-treated animals, with a significant reduction in the heart and kidney. Individual treatments do not significantly alter MDA levels. However, combined treatment significantly improves SOD, GPX, and CAT activities in the heart and kidney, indicating a superior protective effect, suggesting potential to mitigate CP-induced toxicity.

The highly selective SGLT2 inhibitor (SGLT2i) is reported to have beneficial effects on diabetic cardiac hypertrophy; however, the molecular mechanisms underlying the cardioprotection of SGLT2i are not fully understood. In this study, we investigated the impact of the SGLT2 Inhibitor empagliflozin (EMPA) on diabetic hearts and its regulatory mechanisms in high-fat-diet (HFD)- and streptozotocin (STZ)-treated rats. Male rats orally administered HFD/STZ treatment for eight weeks, with or without EMPA (10 mg/kg), were used as our in vivo model. Hematoxylin and eosin (H&E) staining was used for histological examination. Western blot analysis and immunohistochemistry were used to analyze the expression of proteins. Daily EMPA administration prevented the HFD/STZ treatment-induced cardiac hypertrophy by activating the AMP-activated protein kinase (AMPK)/transcription factor EB (TFEB)-mediated upregulation of autophagy- and antioxidant-related proteins. Moreover, EMPA treatment decreased oxidative stress by increasing the antioxidant capacity and protein expression of antioxidant proteins while downregulating the levels of 4-hydroxy-2E-nonenal in the hearts of diabetic rats. Furthermore, EMPA treatment decreased cardiomyocyte apoptosis and increased heart mitochondrial function. The AMPK/TFEB signaling-mediated increase in autophagy, antioxidant capacity, mitochondrial function, and attenuated cardiomyocyte apoptosis may be crucial in the anti-hypertrophic effect conferred by SGLT2i. Our clinical implications suggest a novel pharmacological approach for treating diabetic cardiomyopathy by modulating autophagy and redox homeostasis.

Empagliflozin beyond antidiabetic effect: Amelioration of cyclophosphamide-induced testicular toxicity in rats: Orchestrating klotho/Nrf-2/PPAR-γ/NF-κB/Bax/Bcl-2 cues.

Cardiovascular diseases (CVD) are the leading cause of premature death and disability. Hypoxic conditions play a central role in the pathophysiology of all CVD. Empagliflozin (EMPA), a sodium-glucose cotransporter 2 (SGLT2) inhibitor used for diabetes mellitus type II therapy, has demonstrated a beneficial role in improving cardiovascular outcomes for patients with heart failure. Our study aimed to assess the cardioprotective effect of EMPA on primary human cardiomyocytes in a chemically induced hypoxia model. The cardioprotective effect of the SGLT2 inhibitor was evaluated through four individual experiments including: (1) evaluating mitochondrial network integrity, (2) determining cell count, (3) metabolomic profiling, and (4) determining alterations in miRNA expression. After 24 h of EMPA treatment, we observed a significant improvement in mitochondrial network complexity, as evidenced by increased branching (p < 0.05) and a reduced number of rod-shaped mitochondria (p < 0.05) in EMPA-treated cells compared to controls. After cobalt treatment, we didn't observe any protective effect of EMPA in cells affected by cobalt in various biological aspects, including miRNA expression, metabolomics, or viability. Although EMPA treatment was not able to propagate beneficial effects in the presence of cobalt, pretreatment of cells with EMPA indicated a potential cardioprotective effect associated with improving mitochondrial morphology.

Duchenne muscular dystrophy (DMD) is a severe X-linked disorder with progressive myofiber degeneration and fibrosis from dystrophin deficiency. Current therapies are largely supportive with limited anti-fibrotic benefit, prompting new strategies. Sodium-glucose cotransporter-2 inhibitors (SGLT2i) show emerging anti-fibrotic and anti-inflammatory effects. Open-access proteomic and transcriptomic data sets were integrated for in silico analyses, including differential gene expression, weighted gene co-expression network analysis, and pathway enrichment, to identify dysregulated pathways potentially reversible by SGLT2i. Immune cell composition was estimated using CIBERSORTx in human and murine data sets. Therapeutic effects were tested with empagliflozin (EMPA) in mdx mice (30 mg/kg per day for 4 weeks, starting at 12 weeks) and DMDmdx rats (10 mg/kg per day for 4 months, starting at 5 months), with vehicle controls. Validation used quantitative RT-PCR, grip-strength testing, and histologic fibrosis staining. Analyses highlighted dysregulated extracellular matrix organization, cytokine signaling, and immune responses. Forty overlapping genes were identified; hub genes included COL3A1, COL5A2, and TGFB1. EMPA reduced Tgfb1 expression in DMD rats and significantly decreased collagen deposition in skeletal muscle. Functional testing showed longer grip duration in EMPA-treated mice. Immune profiling revealed shifts in T cells and macrophages, indicating immunomodulation. Findings were consistent across species and data modalities analyzed. These results demonstrate that EMPA modulates fibrosis, inflammation, and muscle endurance in DMD models. These data support repurposing SGLT2i as a promising therapeutic strategy for DMD.

It is known that the daily feeding cycle affects the dosing time-dependent changes in the pharmacodynamics and pharmacokinetics of many drugs. Our previous study demonstrated that administration of empagliflozin (EMPA), sodium-glucose cotransporter 2 (SGLT2) inhibitor, at the beginning of daily feeding cycle (active phase) effectively prevents the development of neuropathic pain in streptozotocin (STZ)-induced diabetic mice. Although the blood glucose levels are closely related to feeding, the relationship between the daily feeding cycle and the optimal dosing time of EMPA remains unclear. In this study, we used STZ-induced diabetic mice and implemented a daily time-restricted feeding (TRF) regimen to investigate whether the dosing time-dependent preventive effect of EMPA on the diabetic neuropathy is modulated by TRF. Animals were housed under a 12-h light/dark cycle, and were assigned to either light-phase TRF (feeding during the light phase) or dark-phase TRF (feeding during the dark phase). The hypoglycemic effect of EMPA was enhanced when the drug was administrated at the beginning of both TRF conditions. A similar influence of the daily feeding cycle on the dosing time-dependent hypoglycemic effect of EMPA was also observed in its preventive effect on the development of diabetic neuropathic pain. Further analysis revealed that dosing time-dependent variations in both the hypoglycemic effect of EMPA and its preventive effect on diabetes-induced pain hypersensitivity were attributable to corresponding changes in urinary glucose excretion. Our results support the notion that the administration of EMPA at the onset of daily feeding cycle effectively suppresses the development of diabetic peripheral neuropathy.

Comparative effects of empagliflozin and vildagliptin on inflammation and atrial function in patients with type 2 diabetes and coronary artery disease: EMPA-VILDA-Response trial.

Abstract Background Systemic inflammation is a hallmark of acute myocardial infarction (AMI) with residual inflammatory risk a key player in reoccurring cardiovascular events. Use of sodium-glucose cotransporter-2 inhibitors (SGLT2i) in patients with type-2 diabetes mellitus (T2DM) and AMI improves cardiovascular outcomes, yet the mechanism of action is unknown. Purpose We posit that Empagliflozin (EMPA) provides anti-inflammatory protection via suppression of the nod-like receptor protein-3 (NLRP3) inflammasome in monocyte-derived macrophages (MDMs), and that this protection is greater in early (Arm A: prescribed EMPA during AMI admission at T0, T30, T90 and T180days) versus delayed therapy (Arm B: EMPA prescribed 90days post-discharge with sampling at T90, T120 and T180days), plus appropriate controls (T0-T90). Methods Patients presenting with T2DM-AMI, eligible but not already taking an SGLT2i were recruited and randomised to Arm A (n=28) or B (n=27). Blood CD14+ monocytes were isolated and differentiated into MDMs. An inflammasome assay measured Caspase-1 activity (direct inflammasome activation) in lipopolysaccharide (0.1ug/mL) + ATP (2mM) stimulated MDMs. Quantitative RT-PCR and Lumit immunoassays measured IL1β, IL6, and TNFα gene expression/secretion. Results IL1β expression (NLRP3 priming) decreased in MDMs isolated from patients in Arm A given EMPA at T0, at all subsequent timepoints from 2.13±0.53 (T0) to 0.64±0.37 (T30; P&amp;lt;0.05), 0.89±0.20 (T90; P&amp;lt;0.05) and 0.66±0.14 (T180; P&amp;lt;0.05). No change in IL1β expression was observed in Arm B both without EMPA (T0-90) or when EMPA was delayed (T90-180). Caspase-1 activity decreased with EMPA in Arm A from 8677±1550 to 4751±591.7 (T0-90; P&amp;lt;0.05) and 3772±450.2 RLu; (T0-180; P&amp;lt;0.01) whilst no significant reduction in caspase-1 activity was recorded in MDMs from Arm B. Although early therapy failed to alter TNFα levels (T0-T180), EMPA prevented the early increase in TNFα gene expression in Arm B MDMs (No EMPA) at T0-T30 (0.537±0.104 to 1.148±0.236 P&amp;lt;0.01). EMPA also reduced TNFα release when taken immediately prior to discharge (from 5240±1201 to 2409±363.6pg/mL; P&amp;lt;0.05; T0-90) compared to Arm B patients without EMPA (T0-90;) or with delayed therapy (T90-180). Lastly, early initiation of EMPA reduced IL6 secretion from 2248± 609.5 (T0) to 883.3±180.4 (T30; P&amp;lt;0.05) and 735.2±0.5pg/mL (T180; P&amp;lt;0.05). In the absence of EMPA, IL6 release increased from 571.9±96.57 (T0) to 1279±243.3 (T30; P&amp;lt;0.05) and 1617±396pg/mL (T90; P&amp;lt;0.05). Levels were restored with EMPA (from 1617±396 to 661.9±82.95pg/mL; P&amp;lt;0.05; T90-180). Conclusion UK NICE guidelines do not stipulate if SGLT2i’s should be prescribed during admission with AMI prior to discharge, or 90days post-discharge. This study suggests that anti-inflammatory protection of EMPA on MDMs from people with T2DM who have had an AMI is greatest following early therapy, timing of which may reduce residual inflammatory risk and improve clinical outcomes.