Boehringer Ingelheim Pharma GmbH Research Articles

Dose–response effects on HbA1c and bodyweight reduction of survodutide, a dual glucagon/GLP-1 receptor agonist, compared with placebo and open-label semaglutide in people with type 2 diabetes: a randomised clinical trial

Aims/hypothesisThe aim of this study was to assess the dose–response effects of the subcutaneous glucagon receptor/glucagon-like peptide-1 receptor dual agonist survodutide (BI 456906) on HbA1c levels and bodyweight reduction.MethodsThis Phase II, multicentre, randomised, double-blind, parallel-group, placebo-controlled study, conducted in clinical research centres, assessed survodutide in participants aged 18–75 years with type 2 diabetes, an HbA1c level of 53–86 mmol/mol (7.0–10.0%) and a BMI of 25–50 kg/m2 on a background of metformin therapy. Participants were randomised via interactive response technology to receive survodutide (up to 0.3, 0.9, 1.8 or 2.7 mg once weekly [qw; dose group (DG) 1–4, respectively] or 1.2 or 1.8 mg twice weekly [DG 5 and 6, respectively]), placebo or semaglutide (up to 1.0 mg qw). Participants and all those involved in the trial conduct/analysis were blinded; the semaglutide arm was open-label. The primary endpoint was absolute change from baseline in HbA1c after 16 weeks’ treatment. The key secondary endpoint was relative change from baseline in bodyweight after 16 weeks’ treatment.ResultsA total of 413 participants were randomised (DG1, n=50; DG2, n=50; DG3, n=52; DG4, n=50; DG5, n=51; DG6, n=50; semaglutide, n=50; placebo, n=60). The full analysis set comprised 411 treated participants (DG6, n=49; placebo, n=59). Adjusted mean (95% CI) HbA1c decreased from baseline (mean ± SD 64.7±9.2 mmol/mol [8.07±0.84%] after 16 weeks’ treatment: DG1 (n=41), −9.92 mmol/mol (−12.27, −7.56; −0.91% [−1.12, −0.69]); DG2 (n=46), −15.95 mmol/mol (−18.27, −13.63; −1.46% [−1.67, −1.25]); DG3 (n=36), −18.72 mmol/mol (−21.15, −16.29; −1.71% [−1.94, −1.49]); DG4 (n=33), −17.01 mmol/mol (−19.59, −14.43; −1.56% [−1.79, −1.32]); DG5 (n=44), −17.84 mmol/mol (−20.18, −15.51; −1.63% [−1.85, −1.42]); DG6 (n=36), −18.38 mmol/mol (−20.90, −15.87; −1.68% [−1.91, −1.45]). The mean reduction in HbA1c was similar with low-dose survodutide (DG2: −15.95 mmol/mol [−1.46%]; n=46) and semaglutide (−16.07 mmol/mol [−1.47%]; n=45). Mean (95% CI) bodyweight decreased dose-dependently up to −8.7% (−10.1, −7.3; DG6, n=37); survodutide ≥1.8 mg qw produced greater bodyweight reductions than semaglutide (−5.3% [−6.6, −4.1]; n=45). Adverse events (AEs) were reported for 77.8% of survodutide-treated participants (mainly gastrointestinal), 52.5% receiving placebo and 52.0% receiving semaglutide.Conclusions/interpretationSurvodutide reduced HbA1c levels and bodyweight after 16 weeks’ treatment in participants with type 2 diabetes. Dose-related gastrointestinal AEs could be mitigated with slower dose escalations.Trial registrationClinicalTrials.gov NCT04153929 and EudraCT 2019-002390-60.FundingBoehringer Ingelheim Pharma GmbH & Co. KG, Ingelheim, Germany.Graphical

749-P: Selection of BI 456906 as a Dual GCGR/GLP-1R Agonist Based on In Vitro Potency and In Vivo Target Engagement Biomarkers

Glucagon (GCG) and glucagon-like peptide 1 (GLP-1) are peptide hormones that pharmacologically regulate bodyweight by increasing energy expenditure and reducing energy intake, respectively. Several dual (GCG receptor [GCGR]/GLP-1 receptor [GLP-1R]) agonistic peptides are in clinical development for obesity. We describe preclinical data leading to selection of BI 456906 as a dual agonist. Functional potencies of the compounds were determined in CHO-K1 cells stably expressing human GCGR and GLP-1R. EC50 for GCGR mediated cAMP increase were 0.52 nM for BI 456906, 0.92 nM for BI 456908 and 0.44 nM for BI 456987. EC50 for GLP-1R engagement were 0.33 nM, 0.61 nM and 1.43 nM, respectively. Upon acute, single dosing to lean mice, engagement of the GCGR and GLP-1R was determined by testing for improvement in oral glucose tolerance (30 nmol/kg) and increase in plasma FGF-21 and liver nicotinamide N-methyltransferase mRNA expression (100 nmol/kg), respectively. Specificity of the biomarkers was confirmed using the selective GLP-1R agonist semaglutide and GLP-1R knockout mice. Bodyweight and glucose lowering (HbA1c) efficacies were investigated in subchronic dosing studies in diet-induced obese (DIO) and diabetic (db/db) mice, respectively. In DIO mice, BI 456906 (30 nmol/kg qd), BI 456908 (30 nmol/kg qd) and BI 456897 (10 nmol/kg qd) achieved bodyweight lowering from baseline of 25%, 27%, and 26%, respectively. In db/db mice, BI 456906 and BI 456908 (10 and 20 nmol/kg qd) substantially lowered HbA1c (0.4-0.6%); no clear effect was observed for BI 456897 (3 and 7 nmol/kg qd). As a balanced GCGR/GLP-1R agonist with robust in vivo efficacy BI 456906 (30 nmol/kg qd) was further characterized, and superior bodyweight lowering (32% vs 27%; p<0.05) was seen versus a maximally effective semaglutide dose (100 nmol/kg qd). This was attributed to an increase in energy expenditure. The selected candidate, BI 456906, is currently in clinical development in people with obesity and NASH. Disclosure R.Augustin: None. L.Thomas: Employee; Boehringer Ingelheim Pharma GmbH&Co.KG. T.Zimmermann: Employee; Boehringer Ingelheim Pharma GmbH&Co.KG. E.Simon: Employee; Boehringer Ingelheim Pharma GmbH&Co.KG. W.Rist: None. I.Uphues: Employee; Boehringer Ingelheim Pharma GmbH&Co.KG. W.Reindl: None. T.Klein: Employee; Boehringer Ingelheim Pharma GmbH&Co.KG. H.Neubauer: None. Funding Zealand Pharma A/S; Boehringer Ingelheim

1711-P: Assessment of Prediabetes in Patients with Morbid Obesity (MO)

The frequency of prediabetes as well as the optimal method to identify glucose abnormalities in morbid obesity (MO) is subject to debate. We, therefore, evaluated the frequency of prediabetes and the optimal method for its respective diagnosis in a large cohort with MO. We included 1.626 patients with MO (age 34±10 years, BMI 44.6± 7.0 kg/m², 74.3% women). All patients without overt type 2 diabetes (T2D) underwent a 75g OGTT. Prediabetes was diagnosed according to ADA criteria. Demographic, cardiovascular risk-markers, insulin levels and renal parameters were assessed. We excluded 421 (25.9%) patients due to T2D. Regarding prediabetes, 339 patients (28.1%) fulfilled the fasting glucose criterion, 240 the 2-hour postprandial criterion (19.9%), and 594 patients the HbA1c criterion (49.2%). 56 patients (4.6%) fulfilled all three criteria, 810 (67.2%) had at least one criterion. 933 (77.4%) patients fulfilled either the fasting glucose or the HbA1c criterion, and 834 (69.2%) either the 2h postprandial or HbA1c criterion. Patient's characteristics are depicted in table 1. The results of our study suggest that fasting glucose or HbA1c are sufficient to diagnose prediabetes. This is particularly of interest since postoperative changes in gut anatomy following bariatric surgery might render a postoperative OGTT invalid for follow-up, whereas fasting glucose or HbA1c are not affected. Disclosure J.Brix: Advisory Panel; Boehringer Ingelheim Pharma GmbH&Co.KG, Eli Lilly and Company, Novo Nordisk, Speaker's Bureau; AstraZeneca, Eli Lilly and Company, Novo Nordisk. V.Parzer: None. S.L.Huber: None. B.Ludvik: Advisory Panel; Eli Lilly and Company, Novo Nordisk, Boehringer Ingelheim Pharma GmbH&Co.KG, Amgen Inc., AstraZeneca, Research Support; Eli Lilly and Company, Novo Nordisk, Boehringer Ingelheim Pharma GmbH&Co.KG, Amgen Inc., AstraZeneca, Speaker's Bureau; Eli Lilly and Company, Novo Nordisk, Boehringer Ingelheim Pharma GmbH&Co.KG, Amgen Inc., AstraZeneca.

789-P: Real-Life Effectiveness of iGlarLixi (Insulin Glargine 100 U/ml and Lixisenatide) in People with Type 2 Diabetes (T2D) according to Prior Insulin Use

Little is known in real life clinical practice about the effectiveness and safety of iGlarLixi in insulin naïve compared to insulin treated people with T2D. In the European REALI pooled database, we analyzed patient-level data of 1317 people with T2D initiating a 24-week iGlarLixi treatment: 718 were insulin naïve (55%) and 598 insulin treated (45%), with a median treatment duration of 2.5 years. Demographics were similar among groups with an overall mean±SD age of 61±9 years and BMI 32.2±5.5 kg/m². Median T2D duration was 8 years in the naïve group and 10 years in the treated group. At Baseline, mean HbA1c was 9.39% in the naïve and 8.76% in the treated (p<0.001). The least square (LS) mean HbA1c change from Baseline to Week 24 (using MMRM) was -1.57% in the naïve and -1.29% in the treated (Fig 1); the LS mean [95% CI] fasting plasma glucose change was -52.5 [-54.9; -50.1] and -50.4 [-53.4; -47.5] mg/dL, respectively. Mean±SD body weight decreased by 1.8±4.6 kg at Week 24, with no difference between groups. Mean iGlarLixi daily dose at Baseline was 14.3U in the naïve and 24.4U in the treated, and reached 30.7U and 36.4U, respectively, at Week 24. Symptomatic hypoglycemia reports and gastrointestinal side effects were low in both groups. This analysis demonstrated effectiveness and safety of iGlarLixi in both insulin naïve and insulin treated people with T2D in clinical practice. Disclosure J.Seufert: Advisory Panel; Abbott Diabetes, Boehringer Ingelheim Pharma GmbH&Co.KG, Sanofi-Aventis Deutschland GmbH, Speaker's Bureau; Abbott Diabetes, Bayer Inc., Boehringer Ingelheim Pharma GmbH&Co.KG, Merck Sharp & Dohme Corp., Sanofi-Aventis Deutschland GmbH, Novartis, Novo Nordisk. N.Freemantle: Advisory Panel; AstraZeneca, Research Support; Galderma, ALK-Abelló A/S, Ipsen Biopharmaceuticals, Inc., Vertex Pharmaceuticals Incorporated, Thea, Speaker's Bureau; Abbott Singapore, ALK Shanghai. C.Guja: Advisory Panel; AstraZeneca, Boehringer-Ingelheim, Eli Lilly and Company, Novo Nordisk A/S, Sanofi, Speaker's Bureau; AstraZeneca, Boehringer-Ingelheim, Eli Lilly and Company, Merck Sharp & Dohme Corp., Novo Nordisk A/S, Sanofi, Servier Laboratories, Viatris Inc. M.Haluzik: Advisory Panel; Novo Nordisk, Lilly Diabetes, Boehringer-Ingelheim, Research Support; Sanofi, Speaker's Bureau; Abbott, AstraZeneca. G.Bigot: None. M.Tournay: None. M.Bonnemaire: Employee; Sanofi, Stock/Shareholder; Sanofi. J.T.Kis: None. Funding Sanofi

318-OR: Tirzepatide Reduces Albuminuria in Patients with T2D—Post Hoc Pooled Analysis of SURPASS 1–5

In SURPASS 4, the GIP/GLP-1 receptor agonist tirzepatide (TZP) showed a potential kidney protective effect in people with T2D and high CV risk by slowing the rate of eGFR decline and reducing urine albumin-creatinine ratio (UACR) vs insulin glargine over 2 years. In this post-hoc analysis, we explored effects of TZP on UACR changes in SURPASS 1-5 trials. UACR (% difference) for TZP (5, 10, 15 mg) vs comparators (COMPs) was analyzed. Analyses were conducted in the pooled entire SURPASS 1-5 population and populations pooled by COMP: placebo (SURPASS 1 & 5); active (SURPASS 2 [semaglutide 1 mg] & SURPASS 3-4 [insulins]); and insulins. In each pooled population, data were examined in all patients and in subgroups defined by baseline UACR ≥30 mg/g or eGFR <60 mL/min/1.73m2. Mixed model for repeated measures was used to analyze on-treatment data from baseline up to the end of treatment visit. UACR data was available in 6263 patients of whom 1846 had UACR ≥30 mg/g and 537 had eGFR <60 mL/min/1.73m2. UACR decreased more with TZP 5, 10, and 15 mg vs COMPs in pooled SURPASS 1-5 and consistently across pooled placebo, active, and insulin COMP studies (Table). UACR lowering appeared more pronounced in subgroups with baseline UACR ≥30 mg/g or eGFR <60 mL/min/1.73m2. In people with T2D, including those with reduced kidney function, TZP decreased UACR vs COMPs to a clinically relevant degree, supporting a potential kidney protective effect. Disclosure H.L.Heerspink: Consultant; AstraZeneca, Boehringer Ingelheim International GmbH, Bayer Inc., Eli Lilly and Company, Chinook Therapeutics Inc., CSL Behring, Gilead Sciences, Inc., George Clinical, Merck & Co., Inc., Janssen Research & Development, LLC, Traveere Pharmaceuticals, Novo Nordisk. K.R.Tuttle: Consultant; Lilly, AstraZeneca, Gilead Sciences, Inc., Research Support; Bayer Inc., Boehringer Ingelheim (Canada) Ltd., Novo Nordisk, Goldfinch Bio, Inc., Traveere Pharmaceuticals. I.Pavo: Employee; Eli Lilly and Company. A.Haupt: Employee; Lilly, Stock/Shareholder; Lilly. Z.Yang: Employee; Eli Lilly and Company. R.Wiese: Employee; Eli Lilly and Company. A.Hemmingway: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. D.Cherney: Other Relationship; Boehringer Ingelheim-Lilly, Merck, AstraZeneca, Sanofi, Mitsubishi-Tanabe, Abbvie, Janssen, Bayer, Prometic, BMS, Maze, Gilead, CSL-Behring, Otsuka, Novartis, Youngene, Lexicon and Novo-Nordisk, Research Support; Boehringer Ingelheim-Lilly, Merck, Janssen, Sanofi, AstraZeneca, CSL-Behring and Novo-Nordisk. N.Sattar: Advisory Panel; Amgen Inc., Boehringer Ingelheim and Eli Lilly Alliance, Novartis, Roche Diagnostics, Consultant; Afimmune Limited, Hanmi Pharm. Co., Ltd., Pfizer Inc., Other Relationship; Abbott, AstraZeneca, Boehringer Ingelheim Pharma GmbH&Co.KG, Eli Lilly and Company, Janssen Pharmaceuticals, Inc., Novo Nordisk, Sanofi, Research Support; AstraZeneca, Boehringer Ingelheim Pharma GmbH&Co.KG, Novartis, Roche Diagnostics. Funding Eli Lilly and Company

797-P: Safety Profile of Once-Weekly Subcutaneous Semaglutide in a Real-World Adult Population with T2D

Real-world (RW) drug safety studies complement safety information from randomized clinical trials. This post hoc analysis of pooled data from nine global non-interventional studies (SURE program: 10 countries, ~30 weeks duration) investigated the safety and prescription patterns of once-weekly (OW) semaglutide in 3,505 adults with T2D in routine practice, overall and by patient subgroup and medical specialty stratification. All safety data reported by site physicians were collected and presented using descriptive statistics. Results: Tables 1 and 2. A two-fold increase in level-2 hypoglycemia was associated with insulin use and microvasculopathy. No new safety concerns were observed with OW semaglutide in this large, diverse, RW population with T2D; the safety profile was in line with that of phase 3 trials of semaglutide. Disclosure J.Yale: Advisory Panel; Sanofi, Novo Nordisk Canada Inc., Eli Lilly and Company, Bayer Inc., Boehringer Ingelheim (Canada) Ltd., Abbott, Mylan, Research Support; Sanofi, Novo Nordisk Canada Inc., Bayer Inc., Speaker's Bureau; Sanofi, Novo Nordisk Canada Inc., Eli Lilly and Company, Bayer Inc., AstraZeneca, Merck & Co., Inc., Boehringer Ingelheim (Canada) Ltd., Janssen Pharmaceuticals, Inc., Abbott, Dexcom, Inc. A.Major-pedersen: Employee; Novo Nordisk A/S. A.Catarig: Employee; Novo Nordisk, Stock/Shareholder; Novo Nordisk. R.Jain: Employee; Novo Nordisk Global Business Services, Stock/Shareholder; Novo Nordisk Global Business Services. M.Menzen: Advisory Panel; Boehringer Ingelheim Pharma GmbH&Co.KG, Novo Nordisk A/S, Ascensia Diabetes Care, Eli Lilly and Company, Novartis, Speaker's Bureau; Abbott Diabetes, AstraZeneca, Santis, Boehringer Ingelheim Pharma GmbH&Co.KG, Novo Nordisk A/S, Bayer Inc., Eli Lilly and Company, Novartis. P.Holmes: Advisory Panel; Boehringer-Ingelheim, Sanofi, Research Support; Novo Nordisk, Roche Diagnostics, Speaker's Bureau; Novo Nordisk, AstraZeneca, Eli Lilly and Company, Bayer Inc. Funding Novo Nordisk A/S

76-LB: Glucagon-Like Peptide-1 Receptor Agonist (GLP-1RA)–Experienced Adults with Type 2 Diabetes (T2D) Switching to Once-Weekly (OW) Semaglutide in a Real-World Setting—SURE Program Post Hoc Analysis

Retrospective database studies have shown that switching between some GLP-1RAs may be associated with additional benefits on glycemic control and body weight (BW). This prospective post hoc analysis of pooled data from the SURE real-world program (9 ~30 week studies in 10 countries) investigated outcomes associated with switching to OW semaglutide from another GLP-1RA in adults with T2D. Endpoints included change in HbA1c and BW from baseline to end of study, percentage achieving HbA1c <7% (when baseline HbA1c ≥7%) or BW change ≥5%, and safety. For change in HbA1c and BW, a random coefficient-adjusted mixed analysis for repeated measures was used, regardless of semaglutide treatment status. In total 651 participants (mean age 60 years; 53% male) switched to OW semaglutide: 64% from liraglutide, 31% from dulaglutide. Significant changes were observed in HbA1c and BW (Figure); 33% of participants with baseline HbA1c ≥7% achieved HbA1c <7%, and 28% reduced BW by ≥5%. No new safety concerns were identified; 3% discontinued due to AEs, 2% reported ≥1 hypoglycemic episode (blood glucose <54 mg/dL) and 8/11 participants with ≥1 level 2 hypoglycemic episode were on insulin. In adults with T2D previously treated with GLP-1RAs, switching to OW semaglutide may provide additional benefits with convenient OW dosing. Disclosure G. Rudofsky: Consultant; AstraZeneca, Eli Lilly and Company, Speaker's Bureau; Abbott Diabetes, Bayer Inc., Boehringer Ingelheim Inc., Eli Lilly and Company, Novo Nordisk, Sanofi. M. Menzen: Advisory Panel; Boehringer Ingelheim Pharma GmbH&Co. KG, Novo Nordisk A/S, Ascensia Diabetes Care, Eli Lilly and Company, Novartis, Speaker's Bureau; Abbott Diabetes, AstraZeneca, Santis, Boehringer Ingelheim Pharma GmbH&Co. KG, Novo Nordisk A/S, Bayer Inc., Eli Lilly and Company, Novartis. L. Potier: Consultant; Lilly Diabetes, Novo Nordisk, Sanofi, AstraZeneca, Boehringer Ingelheim Inc. A. Catarig: Employee; Novo Nordisk, Stock/Shareholder; Novo Nordisk. A. Clark: Employee; Novo Nordisk A/S. P. Priyadarshini: None. C. Abreu: Speaker's Bureau; Lilly, Novo Nordisk. Funding Novo Nordisk

1237-P: Proteomic Indicators of Metabolic Health in Diabetes and Social Vulnerability

Introduction: Understanding the health impacts of socioeconomic deprivation (SED) and its interaction with type 2 diabetes is important for patient care and effective public health initiatives. Large-scale proteomic profiling using aptamer-based technology to measure 7,000 proteins has facilitated the development of blood-based proteomic signatures for 10 cardiometabolic SomaSignal Tests (SST), including a test that predicts the 4-year risk of a cardiovascular event (CV risk). Methods: The cross-sectional PsoBid study recruited participants from socioeconomically deprived and affluent areas of Glasgow, Scotland with similar age ranges and equal proportions of male and female, to identify relevant health disparities. We used regression analyses to estimate the association between SSTs and socioeconomic status (affluent (n = 257, 50.7%), deprived (n = 250, 49.3%)) and prevalent diabetes (n = 27 (5.3%), 7 (2.7%) affluent, 20 (8.0%) SV). Results: Diabetes was associated with SST predicted increased visceral fat, lean body mass, liver fat, body fat percentage, resting energy rate, and CV risk score, and worse proteomic predictors of cardiorespiratory fitness, glucose tolerance, alcohol impact, and kidney health (p<0.05). SED was associated with SST predicted increased visceral adiposity and CV risk score, and decreased cardiorespiratory fitness, glucose tolerance, and kidney health (p<0.05). The association between diabetes and CV risk score was highest among socioeconomically deprived participants (interaction p-value: 0.01). Conclusion: These results demonstrate that participants with diabetes and SED have proteomic phenotypes consistent with decreased cardiometabolic health compared to participants without diabetes or SV and SED is associated with worse predicted cardiovascular risk in patients with diabetes. Proteomics may be useful in identifying and monitoring the health effects of diabetes and SED or the impact of relevant interventions. Disclosure M.Simpson: Employee; SomaLogic Operating CO., Inc. R.Ostroff: Employee; SomaLogic. S.A.Williams: Employee; SomaLogic. N.Sattar: Advisory Panel; Amgen Inc., Boehringer Ingelheim and Eli Lilly Alliance, Novartis, Roche Diagnostics, Consultant; Afimmune Limited, Hanmi Pharm. Co., Ltd., Pfizer Inc., Other Relationship; Abbott, AstraZeneca, Boehringer Ingelheim Pharma GmbH&Co.KG, Eli Lilly and Company, Janssen Pharmaceuticals, Inc., Novo Nordisk, Sanofi, Research Support; AstraZeneca, Boehringer Ingelheim Pharma GmbH&Co.KG, Novartis, Roche Diagnostics.

794-P: Effectiveness and Safety of iGlarLixi in People with Type 2 Diabetes (Pw2D), Not at Target on Basal Insulin (BI) and Oral Antidiabetic Therapy (BOT)—Results from the Observational, Prospective Study CHANCE

iGlarLixi, a fixed-ratio combination of insulin glargine 100 U/mL and lixisenatide, is a once-daily treatment advancement option for Pw2D not at target on BOT. CHANCE investigated effectiveness and safety of switching the BI to iGlarLixi in daily clinical practice. Primary objective was to assess absolute change in HbA1c 24 weeks (W24) after switching to iGlarLixi. Secondary objectives included changes in fasting plasma glucose (FPG), bodyweight (BW), use of oral antidiabetic drugs (OADs), dose of previous BI, and hypoglycemia incidence. Overall, data from 70 Pw2D were analyzed. Mean BI dose was 38.9 units per day and individual target HbA1c was 6.9%. iGlarLixi was started with 30 dose steps per day (DS/d); at W24, mean dose was 41.6 DS/d. HbA1c decreased by -0.7% (P < 0.001), while FPG trended down by ~20 mg/dL. This was accompanied by a -3.0 kg decrease in BW (P = 0.442). Numbers of OADs used decreased markedly 24 weeks after switching to iGlarLixi (P < 0.05). Hypoglycemia incidence was low before and did not change significantly after switching to iGlarLixi. In conclusion, advancing BOT by switching the BI to iGlarLixi, improved glycemic control of Pw2D in daily clinical practice without BW gain and with low hypoglycemia incidence; however, titration was not sufficient to reach glycemic targets after 24 weeks. Disclosure T.Wiesner: Advisory Panel; Roche Diabetes Care, Sanofi-Aventis Deutschland GmbH, Speaker's Bureau; Sanofi-Aventis Deutschland GmbH, Lilly Diabetes, Boehringer Ingelheim Inc., Abbott Diabetes, Novo Nordisk, Dexcom, Inc., Novartis. M.Pfohl: Advisory Panel; Boehringer Ingelheim and Eli Lilly Alliance, Sanofi-Aventis Deutschland GmbH, Speaker's Bureau; Sanofi-Aventis Deutschland GmbH. K.Pegelow: Employee; Sanofi-Aventis Deutschland GmbH, Stock/Shareholder; Sanofi-Aventis Deutschland GmbH. J.Müller: Employee; Sanofi-Aventis Deutschland GmbH, Stock/Shareholder; Bayer Inc., Inmunebio Inc. J.Seufert: Advisory Panel; Abbott Diabetes, Boehringer Ingelheim Pharma GmbH&Co.KG, Sanofi-Aventis Deutschland GmbH, Speaker's Bureau; Abbott Diabetes, Bayer Inc., Boehringer Ingelheim Pharma GmbH&Co.KG, Merck Sharp & Dohme Corp., Sanofi-Aventis Deutschland GmbH, Novartis, Novo Nordisk. Funding Sanofi-Aventis Deutschland GmbH

407-P: Efficacy of Finerenone in Patients with Abnormal Markers of Liver Steatosis and Fibrosis—A FIDELITY Subgroup Analysis

Introduction: Finerenone, a selective, nonsteroidal mineralocorticoid receptor antagonist, showed cardiorenal benefits vs placebo in patients with chronic kidney disease and type 2 diabetes. Here, we analyzed patients with liver function abnormalities in FIDELITY, a prespecified pooled dataset from the FIDELIO-DKD and FIGARO-DKD trials. Methods: Patients with urine albumin-to-creatinine ratio ≥30-≤5000 mg/g and estimated glomerular filtration rate (eGFR) ≥25 mL/min/1.73 m2 and type 2 diabetes were analyzed according to elevated transaminases, high risk of steatosis (hepatic steatosis index [HSI] >36), and risk of intermediate to advanced fibrosis (Fibrosis-4 Index [FIB-4] scores >1.30, >2.67, and >3.25). Changes in liver function and in kidney (kidney failure, sustained 57% eGFR decline, or renal death) and cardiovascular (CV; CV death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for heart failure) composite outcomes were assessed. Results: Across 13,026 patients, finerenone reduced the risk of kidney and CV outcomes compared with placebo in patients with elevated transaminases (HR=0.75; 95% CI 0.50-1.13 and HR=0.81; 95% CI 0.62-1.07, respectively), at high risk of steatosis (HR=0.75; 95% CI 0.64-0.88 and HR=0.85; 95% CI 0.77-0.95, respectively), and at high risk of intermediate fibrosis (HR=0.75; 95% CI 0.60-0.93 and HR=0.76; 95% CI 0.67-0.87, respectively), with stronger reductions in the risk of the CV composite outcome at higher FIB-4 scores (HR=0.61; 95% CI 0.41-0.92; p-value for interaction=0.13 and HR=0.48; 95% CI 0.25-0.90; p-value for interaction=0.03 for FIB-4 >2.67 and >3.25, respectively). Liver transaminase levels remained consistent between treatment groups throughout the study. Conclusions: Finerenone demonstrated robust and consistent cardiorenal benefits in patients with abnormal liver function and even more profound CV benefits in patients with higher FIB-4 scores. Disclosure N.Perakakis: Advisory Panel; Bayer Inc., Other Relationship; Novo Nordisk, Novo Nordisk. P.Kolkhof: Employee; BAYER AG. R.Lawatscheck: Employee; Bayer Inc. C.Scott: None. G.Bakris: Advisory Panel; Janssen Pharmaceuticals, Inc., Consultant; AstraZeneca, Boehringer Ingelheim (Canada) Ltd., Other Relationship; Bayer Inc. S.R.Bornstein: None. A.L.Birkenfeld: None. A.Linkermann: None. S.Anker: Advisory Panel; AstraZeneca, Novartis AG, Consultant; Bayer Inc., Boehringer Ingelheim Pharma GmbH&Co.KG, Vifor Pharma Management Ltd., Research Support; Abbott, Vifor Pharma Management Ltd. G.Filippatos: Other Relationship; Boehringer-Ingelheim, Bayer Inc., Amgen Inc., Medtronic, Vifor Pharma Management Ltd., Novartis, Servier Laboratories. B.Pitt: Consultant; Bayer Inc., AstraZeneca, Boehringer Ingelheim International GmbH, Vifor Pharma Management Ltd., Lexicon Pharmaceuticals, Inc., PhaseBio Pharmaceuticals, Inc., KBP Bioscience, Merck & Co., Inc. P.Rossing: Other Relationship; Abbott Diagnostics, AstraZeneca, Bayer Inc., Boehringer Ingelheim Inc., Novo Nordisk, Merck KGaA, Gilead Sciences, Inc., Sanofi. L.M.Ruilope: Advisory Panel; Bayer Inc.

829-P: Outcomes with Finerenone in People with CKD and T2D by Baseline Insulin Resistance—A FIDELITY Subgroup Analysis

Introduction Insulin resistance (IR) is associated with increased risk of T2D, CV disease and CKD. Finerenone, a nonsteroidal mineralocorticoid receptor antagonist, has shown cardiorenal benefits in people with CKD and T2D. We investigated if IR is associated with risk of cardiorenal outcomes and if IR modifies finerenone efficacy in the pooled FIDELITY dataset. Methods People with T2D and CKD (UACR ≥30-<300 mg/g + eGFR ≥25-≤90 mL/min/1.73 m2 or UACR ≥300-≤5000 mg/g + eGFR ≥25 mL/min/1.73 m2) and optimized RAS blockade were randomized to finerenone or placebo. Estimated glucose disposal rate (eGDR), an inverse marker of IR, was calculated by 21.158 + (-0.09 × waist circumference [cm]) + (-3.407 × presence of hypertension) + (-0.551 × HbA1c [%]). Risk of CV (time to CV death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for heart failure) and kidney (time to kidney failure, sustained ≥57% eGFR decline, or renal death) composite outcomes, and safety, were assessed by eGDR. Results Median baseline eGDR was 4.1 mg/kg/min in 12,964 participants. Analysis by continuous baseline eGDR showed a significantly lower risk of CV events at 3.5 years with increasing eGDR (HR 0.88 [95% CI 0.86-0.91] p<0.01); whereas eGDR was not associated with kidney outcomes. Analyses by eGDR subgroups showed no significant heterogeneity for the effect of finerenone on CV (eGDR <median: HR 0.82 [95% CI 0.72-0.92]; eGDR ≥median: HR 0.91 [95% CI 0.78-1.06]; p-interaction=0.29) or kidney outcomes (eGDR <median: HR 0.85 [95% CI 0.70-1.04]; eGDR ≥median: HR 0.69 [95% CI 0.57-0.84]; p-interaction=0.18). Consistent strength and direction of the associations were observed across sensitivity analyses. Adverse events were similar across eGDR subgroups; hyperkalaemia events were higher with finerenone than placebo. Conclusions A higher risk of CV, but not kidney, outcomes was observed with greater IR in people with T2D and CKD. Finerenone efficacy and safety were not modified by baseline IR. Disclosure T.Ebert: Advisory Panel; Sanofi, Bayer Inc., Research Support; AstraZeneca, Speaker's Bureau; CME-Verlag, Santis, Fresenius Medical Care. K.Rohwedder: Employee; Bayer Inc., AstraZeneca. P.Rossing: Other Relationship; Abbott Diagnostics, AstraZeneca, Bayer Inc., Boehringer Ingelheim Inc., Novo Nordisk, Merck KGaA, Gilead Sciences, Inc., Sanofi. S.Anker: Advisory Panel; AstraZeneca, Novartis AG, Consultant; Bayer Inc., Boehringer Ingelheim Pharma GmbH&Co.KG, Vifor Pharma Management Ltd., Research Support; Abbott, Vifor Pharma Management Ltd. L.M.Ruilope: Advisory Panel; Bayer Inc. P.Fioretto: Advisory Panel; Eli Lilly and Company, Novo Nordisk, Speaker's Bureau; AstraZeneca, Boehringer-Ingelheim, Bayer Inc. V.Fonseca: Consultant; Abbott, Corcept Therapeutics, Eli Lilly and Company, Other Relationship; BRAVO4HEALTH, LLC, Research Support; Fractyl Health, Inc., Stock/Shareholder; Amgen Inc. G.Umpierrez: Research Support; Abbott, Dexcom, Inc., Baxter. A.L.Birkenfeld: None. R.Lawatscheck: Employee; Bayer Inc. C.Scott: None.

69-LB: Sex-Specific Dynamics of GLP-1R and GIPR Expression in the Hypothalamus and the Amygdala of Mice

GIPR:GLP-1R co-agonism improves obesity and cardio-metabolic disease with superior efficacy in both male and female mice compared to mono-agonism. We detected sex-specific differences in metabolic phenotypes, which may point to differing sensitivity for GLP-1R and/or GIPR in areas of the hypothalamus regulating energy balance (ARC, VMH, DMH) and in areas of the amygdala regulating cardiovascular functions. To optimize future therapeutic benefits and minimize side effects from a perspective of sex differences, we investigated whether GLP1R and GIPR expression levels in specific cell populations of the hypothalamus and the amygdala regions are co-determined by sex and diet. Female and male C57Bl6 mice were fed normal chow or 58% high fat diet (HFD) for 21 days (n = 4, total mouse number 16). Spatial transcriptomics by in-situ hybridization method RNAscope™ was combined with immunofluorescence staining for neurons (NeuN), astrocytes (GFAP) and microglia (iba1) to identify cell type specific gene expression patterns of GLP1R and GIPR in female and male brains. In the hypothalamus, both sexes, gene expression of GLP-1R was higher compared to GIPR (ARC, VMH and DMH). In our studies GLP-1R and GIPR were mostly expressed in neurons, marginally in microglia and not observed in astrocytes in both sexes. HFD exposure reduced GLP-1R and GIPR gene expression in males only. In the Amygdala, neuronal GIPR gene expression levels were higher compared to GLP-1R in both sexes. HFD reduced GIPR and GLP-1R gene expression in males only. In females, HFD induced a slight increase of GIPR gene expression specifically in the central and basolateral amygdala. Our results reveal sex-specific dynamics in GLP-1R and GIPR expression patterns in metabolically relevant areas of the hypothalamus and the amygdala induced by HFD exposure. This may imply sex-specific dosing regimens of GIP:GLP-1R co-agonists for future treatments of cardio-metabolic disease Disclosure S. M. Hofmann: None. S. Zhang: None. R. Tom: None. T. D. Müller: Research Support; Novo Nordisk A/S, Speaker's Bureau; Eli Lilly and Company, AstraZeneca, Mercodia AB, Stock/Shareholder; Novo Nordisk A/S, Eli Lilly and Company. M. H. Tschöp: Consultant; Boehringer Ingelheim Pharma GmbH&Co. KG.

1566-P: The TM6SF2 rs58542926 Polymorphism Dissociates Hepatocellular Lipids from Insulin Resistance in Recent-Onset Type 2 Diabetes

Increased hepatocellular lipid content (HCL) is generally linked to insulin resistance, which contributes to greater risk of type 2 diabetes and cardiovascular complications. Conversely, a single-nucleotide polymorphism (TM6SF2EK; rs58542926) in the transmembrane 6 superfamily member 2 gene has been associated with nonalcoholic fatty liver disease (NAFLD), but lower cardiovascular risk. Thus, this case-control study tested the role of this polymorphism for tissue-specific insulin sensitivity during the early course of diabetes. Males with recent-onset type 2 diabetes with (TM6SF2EK: n=16) or without (TM6SF2EE: n=16) the heterozygous TM6SF2 polymorphism of similar age and BMI, underwent Botnia-clamps with [6,6-2H2]glucose to measure whole-body, hepatic and adipose tissue insulin sensitivity. HCL and liver fibrosis were assessed with 1H-magnetic resonance spectroscopy and non-invasive tests (FIB4, APRI, AST/ALT), respectively. A subset of both groups (n=24) was re-evaluated after 5 years. Despite doubled HCL, TM6SF2EK had similar hepatic and adipose tissue insulin sensitivity and even 27% higher whole-body insulin sensitivity than TM6SF2EE. The novel diabetes endotypes were equally distributed among both groups. After 5 years, whole-body insulin sensitivity, HCL and liver fibrosis indices were similar, while adipose tissue insulin sensitivity decreased by 87% and 55% both in TM6SF2EK and TM6SF2EE and circulating triacylglycerol increased in TM6SF2EE only. The TM6SF2 gene polymorphism rs58542926 dissociates ectopic fat content from insulin resistance in recent-onset type 2 diabetes, which is however attenuated by disease duration. This suggests that diabetes-related metabolic alterations dominate over the effects of the TM6SF2 gene polymorphism during the early course of diabetes and NAFLD. Disclosure K.Bódis: None. R.Guthoff: Other Relationship; Alimera, Bayer Inc., Novartis, Allergan. V.Schrauwen-hinderling: None. H.Al-hasani: None. V.Burkart: None. J.Szendroedi: None. R.Wagner: Advisory Panel; Daiichi Sankyo, Speaker's Bureau; Novo Nordisk, Sanofi. D.F.Markgraf: Employee; Boehringer Ingelheim Pharma GmbH&Co.KG. M.Roden: Advisory Panel; Eli Lilly and Company, Consultant; TARGET PharmaSolutions, Inc., Research Support; Boehringer-Ingelheim, Novo Nordisk, Novartis, Sanofi. M.Bombrich: None. M.Schön: None. B.Knebel: None. O.P.Zaharia: None. G.J.Bönhof: None. Y.Karusheva: None. Y.Kupriyanova: None. J.Kotzka: None.

777-P: Early Weight Loss and Late Metabolic Outcomes in Tirzepatide-Treated Participants with Type 2 Diabetes

Tirzepatide (TZP), a novel, once-weekly GIP/GLP-1 receptor agonist, significantly improved glycemic control with robust weight loss vs comparators in the SURPASS 1-5 trials. The aim of this analysis was to assess whether the magnitude of early weight loss with TZP (<5% or ≥5% at Week 8) would be associated with different clinical response in terms of HbA1c and weight at Week 40/42 across the SURPASS 1-5 trials. These phase 3 trials included participants with type 2 diabetes receiving TZP as monotherapy or add-on to background antihyperglycemic medications. This post-hoc analysis used pooled data from all TZP arms (5, 10, and 15 mg) across trials. Participants achieving ≥5% early weight loss at Week 8 had lower HbA1c and weight at baseline, and experienced greater reductions in HbA1c and weight at Week 40/42 vs those achieving <5% early weight loss (Table). Among participants achieving ≥5% early weight loss, greater proportions met HbA1c <7% and ≤6.5%, ≥10% weight loss, and the composite endpoint of HbA1c <7% and ≥10% weight loss. Early weight loss with TZP predicts greater reductions of HbA1c and weight at Week 40/42. However, even the one third of participants achieving <5% weight loss early in the course of therapy are likely to meet the ADA/EASD dual treatment goals of HbA1c <7% and ≥10% weight loss. Disclosure I.Lingvay: Advisory Panel; Novo Nordisk A/S, Lilly Diabetes, Boehringer-Ingelheim, Sanofi, Consultant; Carmot Therapeutics, Inc., Merck Sharp & Dohme Corp., Janssen Scientific Affairs, LLC, Pfizer Inc., Intercept, Intarcia, Valeritas, TargetRWE, Shionogi, Zealand Pharma, Structure, Bayer, Research Support; Novo Nordisk A/S, Boehringer-Ingelheim. F.Giorgino: Advisory Panel; Boehringer-Ingelheim, Amarin Corporation, Medtronic, Roche Diabetes Care, Sanofi, Bayer Inc., Novo Nordisk, Consultant; Novo Nordisk, Lilly, Research Support; Lilly, Roche Diabetes Care, AlfaSigma, Speaker's Bureau; Abbott, Boehringer-Ingelheim, Lilly, Sanofi, Medscape. L.Van gaal: Advisory Panel; Merck Sharp & Dohme Corp., Speaker's Bureau; Boehringer Ingelheim Pharma GmbH&Co.KG, Eli Lilly and Company, Novo Nordisk. M.Liu: Consultant; Eli Lilly and Company. Á.Rodríguez: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. A.Torcello-gomez: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. J.I.Kiljanski: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. Funding Eli Lilly and Company

164-LB: Durable Improvement in HbA1c in Youth-Onset T2D—A Post Hoc Analysis of the DINAMO Trial of Empagliflozin and Linagliptin vs. Placebo

The increasing prevalence of childhood overweight and obesity has led to a substantial increase in T2D in youth. The clinical course of youth-onset T2D is more aggressive than in adults, with a faster decline in beta‑cell function leading to worse glycemic control and an early onset of complications. The DINAMO trial compared the efficacy and safety of empagliflozin and linagliptin with placebo in 157 participants aged 10-17 years. Participants were randomized (1:1:1) to linagliptin 5 mg, empagliflozin 10 mg, or placebo. At Week 12, those in the empagliflozin group who did not attain HbA1c <7.0% were re‑randomized (1:1) to empagliflozin 10 or 25 mg. Responders were defined as having no rescue medication and a HbA1c change from baseline ≤-0.5% or attained HbA1c <7.0% at Week 26. Empagliflozin results were pooled. Significantly more participants responded to empagliflozin vs placebo (rate difference 23.6, 95% CI 4.3-41.4; p=0.01); this response was maintained to Week 52 by 40% of the group. Results for linagliptin vs placebo were not significant. Empagliflozin provided a clinically meaningful response even at higher HbA1c and FPG levels (Table). These results show that empagliflozin provides a clinically relevant, statistically significant, and durable improvement in glycemic control, suggesting SGLT2 inhibitors could be a new therapy for T2D in youth. Disclosure L. M. Laffel: Advisory Panel; Medtronic, Lilly Diabetes, Novo Nordisk, Vertex Pharmaceuticals Incorporated, Roche Diagnostics, Provention Bio, Inc., Consultant; Dexcom, Inc., Janssen Pharmaceuticals, Inc., Medscape. T. Danne: Research Support; Abbott Diabetes, AstraZeneca, Bayer Inc., Boehringer-Ingelheim, Dexcom, Inc., Eli Lilly and Company, Novo Nordisk A/S, Sanofi, Medtronic, Stock/Shareholder; DreaMed Diabetes, Ltd. W. V. Tamborlane: None. G. J. Klingensmith: Advisory Panel; Boehringer Ingelheim Inc., Consultant; AstraZeneca, Boehringer Ingelheim Inc., Novo Nordisk, Takeda Pharmaceutical Co., Ltd. D. Neubacher: Employee; Boehringer Ingelheim Pharma GmbH&Co. KG. I. Tartakovsky: Employee; Boehringer Ingelheim International GmbH. J. Marquard: Employee; Boehringer Ingelheim Inc. P. Zeitler: Consultant; Eli Lilly and Company, Boehringer Ingelheim Inc., Johnson & Johnson. S. M. Willi: Advisory Panel; Boehringer Ingelheim International GmbH, Medtronic. Funding Boehringer Ingelheim and Eli Lilly and Company Diabetes Alliance

Pharmacokinetic and bioequivalence study of Telzap® Plus fixed-dose combination compared with MikardisPlus® in healthy volunteers after single administration

Introduction. A fixed dose combination of telmisartan and hydrochlorothiazide is indicated for treatment of in the treatment of arterial hypertension. The combination of these substances causes an additive effect that helps to reduce blood pressure. A bioequivalence study of Telzap® Plus compared with MikardisPlus® was conducted with 63 volunteers.Aim. The purpose of the bioequivalence trial was a comparative study of the pharmacokinetics and evidence of the bioequivalence of the fixed dose combination drug product Telzap® Plus (tablets 80 mg + 12,5 mg, Zentiva KS company, Czech Republic) compared with drug products MikardisPlus® (telmisartan+hydrochlorothiazide, tablets 80 mg + 12,5 mg, Boehringer Ingelheim Pharma GmbH & Co. KG, Germany) in healthy volunteers after a single administration under fasting.Materials and methods. To prove bioequivalence, an open label, comparative, randomized, crossover fourperiod replicate single-center clinical trial was conducted. The concentrations of hydrochlorothiazide and telmisartan in plasma samples were determined with a validated HPLC-MS/MS method. A pharmacokinetic and statistical analysis was performed and confidence intervals for the pharmacokinetic parameters Cmax and AUC0-72 were calculated.Results and discussion. It can be concluded that the studied formulations are bioequivalent in terms of pharmacokinetic parameters of hydrochlorothiazide and telmisartan. All 90 % confidence intervals for the estimated pharmacokinetic parameters of hydrochlorothiazide were in the range of 80–125 %, 90 % confidence intervals for telmisartan were within the bioequivalence range of 80–125 % for AUC0-72, and 79,30–126,11 % for Cmax.Conclusion. Thus, according to the criteria used in the study, the formulations are proved to be bioequivalent.

Upregulation of SGLT1 and 2 promotes oxidative stress in right atrial appendages of patients with low-grade inflammatory responses: potential role in atrial fibrillation

Abstract Introduction Atrial fibrillation (AF), the most common cardiac arrhythmia, is associated with increased morbidity and mortality. Inflammatory infiltrates have been observed in atrial tissues from AF patients suggesting that low-grade inflammation contributes to the pro-fibrotic, pro-remodeling and pro-senescence responses of the atrial tissue favoring AF. Although sodium glucose co-transporter2 inhibitors (SGLT2i) have shown beneficial effects in heart failure, the expression and role of SGLT2 in cardiac tissues remain unclear. Purpose This study examined the expression of SGLT1 and 2 in right atrial appendages (RAA) of cardiac patients, and, if so, to determine the underling mechanism and their functional role. Methods RAA were harvested from patients undergoing coronary artery bypass surgery or aortic valve replacement at the University Hospital of Strasbourg. Patients with comorbidities such as malignancies and chronic inflammatory diseases were excluded. The expression level of markers was assessed using RT-qPCR, Western blot analysis, and immunofluorescence staining, and the level of reactive oxygen species using dihydroethidium staining. Results Analysis of RAA samples evidenced up-to a 4-fold difference in the NOS3 mRNA level. Those with low levels of NOS3 mRNA showed high levels of ICAM1, F3, MMP9, TGF-β1, TP53, CDKN1A, CDKN2A, SLCA1, SLCA2, IL1B, IL6, TNF-a and CD68 whereas the contrary was observed in those with high levels of NOS3 mRNA. SGLT1/2 mRNA levels were negatively correlated with that of eNOS and positively with those of IL-1, IL-6, TNF- α, CD68, and the senescence marker p53. Western blot analysis of RAA indicated that those expressing low levels of eNOS protein showed high levels of ICAM-1, VCAM-1, TF, MMP-2, TGF-b, SGLT1, SGLT2, p-p65, p53, p21 and p16 proteins whereas the contrary was observed in those with high levels of eNOS protein. Immunofluorescence staining indicated that SGLT1/SGLT2 signals were colocalized with those of CD31, an endothelial cell marker, in RAA, and associated with those of CD68 and TNF-α. Samples with high levels of SGLT1 and 2 displayed higher levels of oxidative stress that were inhibited by the antioxidant NAC (N-acetylcysteine), VAS-2870 (NADPH oxidase inhibitor), perindoprilat (ACE inhibitor), losartan (AT1R antagonist), empagliflozin (selective SGLT2 inhibitor), sotagliflozin (dual SGLT1/2 inhibitor) and by infliximab (TNF-α receptor neutralizing antibody). Conclusion The findings indicate that high expression levels of SGLT1 and 2 are observed in RAA showing low eNOS levels associated with pro-inflammatory, pro-senescent, pro-remodeling and pro-fibrotic responses. They were further associated with increased levels of oxidative stress sensitive to inhibitors of either the local angiotensin system, TNF-alpha or SGLT2. Thus, inhibition of SGLT2 appears to be an interesting approach to reduce the stimulatory pro-oxidant signal in atrium related to low-grade inflammation. Funding Acknowledgement Type of funding sources: Private company. Main funding source(s): Boehringer Ingelheim Pharma GmbH & Co. KG

Empagliflozin treatment differentially modifies visceral and subcutaneous adipose tissue lipidomes and pro-inflammatory cytokines in zucker fatty diabetic rats

Abstract EMPA-REG OUTCOME (Empagliflozin, Cardiovascular Outcome Event Trial) trial highlighted the relevance of pharmacological inhibition of sodium-glucose cotransporter 2 (SGLT2) for the treatment of patients with type 2 diabetes mellitus (T2DM) and/or cardiovascular disease. Although the pathways through which SGLT2 inhibitors exert a beneficial effect on the cardiovascular system are still unknown, it has been suggested that energy metabolism regulation and a reduction of systemic inflammation could be some of the mechanisms implicated. Available data also suggests that empagliflozin treatment could be able to exert regulatory effects on adipose tissue (AT) depots. The aim of our study was to evaluate the impact of empagliflozin treatment on the lipidome of visceral (VAT) and subcutaneous adipose tissue (SAT) depots in a rat model of obesity and T2DM. Diabetic obese Zucker Fatty (ZDF) rats were treated with 30 mg/kg/day of empagliflozin p.o for 6 weeks. The lipidomes of VAT and SAT depots were analyzed using ultra-high performance liquid chromatography coupled to mass spectrometry. Empagliflozin's effect on pro-inflammatory markers in AT was analyzed by RT-PCR. In VAT, 18 metabolites were significantly altered in empagliflozin-treated rats vs. controls. Nearly all diglycerides tested (13 of 14) were significantly increased in treated rats, as the most notable altered chemical class. Furthermore, 3 oxidized fatty acids (FA) and FA like gadoleic acid and linoleic acid were also significantly increased. In SAT, a total of 14 metabolites were significantly altered. Most of them (13 of 14) were glycerophospholipids. Significantly lower levels of 4 lysophosphatidylethanolamines, 4 lysophosphatidylcholines, and 3 lysophosphatidylinositols and higher levels of 2 phosphatidylcholines were shown. In contrast to VAT, a significant decrease in most of these metabolites was observed in empagliflozin-treated samples. Several ratios of metabolites were also calculated to infer the potential enzyme activities related to lipid metabolism in both VAT and SAT. However, the ratios studied were only statistically significant in VAT of empagliflozin-treated rats vs. control, where the main potential enzyme activities altered were desaturases and elongases. Empagliflozin treatment also reduces the expression of the pro-inflammatory cytokines interleukin-1 beta (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNFα) and monocyte-chemotactic protein-1 (MCP-1) in VAT, with no changes in SAT, except for interleukin-13 (IL-13), which also decreases compared to untreated diabetic ZDF rats. In conclusion, empagliflozin increased oxidized FA and diglycerides in VAT and decreased glycerophospholipid levels in SAT. The anti-inflammatory effect of empagliflozin in VAT is not observed in SAT. The effect of empagliflozin on the regulation of the adipose tissue lipidome and inflammatory profile is different depending on the localization of the depots. Funding Acknowledgement Type of funding sources: Private company. Main funding source(s): Boehringer Ingelheim Pharma GmbH and Co

SGLT1/2 expression in the heart of transplanted patients is associated with inflammatory induction of oxidative stress: potential protective effect of SGLT2 inhibitors

Abstract Introduction Heart transplantation is a life-saving surgical procedure for patients with end-stage cardiac dysfunction, however, such procedures are usually at risk of rejection due to acute inflammatory activation. While such inflammatory induction leaves the newly transplanted heart at risk of functional and structural remodeling subsequent to oxidative damage, current anti-inflammatory treatment options expose the patient to an elevated risk for adverse reactions in addition to absence of cardio-protective effects. Therefore, novel therapies with anti-inflammatory and cardio-protective dual effects are needed. Recently, our group has reported that low-grade inflammation is associated with upregulation of SGLT1/2 in arteries of human with cardiovascular diseases. Yet, the role and function of SGLT2 in human cardiac tissue remains poorly understood. Aim This study focuses on the expression pattern of SGLT1/2 in cardiac biopsies of heart transplanted patients and aim to identifying their functional impact. Methods Routine endomyocardial Biopsies (23) were performed for the detection of acute rejection heart transplanted patients (less than 2 years) at our University Hospital. Gene expression levels were assessed using RT-qPCR, the in situ tissue localization of proteins by immunofluorescence staining, and the level of oxidative stress by dihydroethidium staining. Results Gene expression analysis revealed strong inflammatory reaction in 5 samples indicated by at least 20-fold higher levels of mRNA of IL1B, IL6, TNFA and CD68 compared to the other 18 samples and concomitant with high expression levels of SLC5A1, SLC5A2, AT1R, CYBA, NCF1, ICAM1, VCAM1, MMP2, MMP9 and TGFB1 in contrast to low levels of NOS3. In addition, increased levels of oxidative stress were observed in the same biopsies, which were diminished by the antioxidant N-acetylcysteine (NAC), NADPH oxidase inhibitor (VAS-2870), TNF-α receptor neutralizing antibody (infliximab), ACE inhibitor (perindoprilat), AT1R antagonist (losartan), dual SGLT1/2 inhibitor (sotagliflozin) and selective SGLT2 inhibitor (empagliflozin) with inhibitory effects reaching up to 80%. Immunofluorescence staining indicated signals for nitro-tyrosine, TNF-alpha, SGLT1 and 2 in several samples. Conclusion These findings indicate that both isoforms SGLT1 and SGLT2 are expressed in the transplanted human heart and suggest a pattern of expression associated with pro-inflammatory response. They further indicate a potential protective effect of SGLT2 inhibitors in transplanted hearts through mitigating oxidative stress and hence providing a possible novel therapy for heart transplantation recipients to preserve the heart function. Funding Acknowledgement Type of funding sources: Private company. Main funding source(s): Boehringer Ingelheim Pharma GmbH & Co. KG

Sleep EEG in young people with 22q11.2 deletion syndrome: A cross-sectional study of slow-waves, spindles and correlations with memory and neurodevelopmental symptoms.

Young people living with 22q11.2 Deletion Syndrome (22q11.2DS) are at increased risk of schizophrenia, intellectual disability, attention-deficit hyperactivity disorder (ADHD) and autism spectrum disorder (ASD). In common with these conditions, 22q11.2DS is also associated with sleep problems. We investigated whether abnormal sleep or sleep-dependent network activity in 22q11.2DS reflects convergent, early signatures of neural circuit disruption also evident in associated neurodevelopmental conditions. In a cross-sectional design, we recorded high-density sleep EEG in young people (6-20 years) with 22q11.2DS (n=28) and their unaffected siblings (n=17), quantifying associations between sleep architecture, EEG oscillations (spindles and slow waves) and psychiatric symptoms. We also measured performance on a memory task before and after sleep. 22q11.2DS was associated with significant alterations in sleep architecture, including a greater proportion of N3 sleep and lower proportions of N1 and REM sleep than in siblings. During sleep, deletion carriers showed broadband increases in EEG power with increased slow-wave and spindle amplitudes, increased spindle frequency and density, and stronger coupling between spindles and slow-waves. Spindle and slow-wave amplitudes correlated positively with overnight memory in controls, but negatively in 22q11.2DS. Mediation analyses indicated that genotype effects on anxiety, ADHD and ASD were partially mediated by sleep EEG measures. This study provides a detailed description of sleep neurophysiology in 22q11.2DS, highlighting alterations in EEG signatures of sleep which have been previously linked to neurodevelopment, some of which were associated with psychiatric symptoms. Sleep EEG features may therefore reflect delayed or compromised neurodevelopmental processes in 22q11.2DS, which could inform our understanding of the neurobiology of this condition and be biomarkers for neuropsychiatric disorders. This research was funded by a Lilly Innovation Fellowship Award (UB), the National Institute of Mental Health (NIMH 5UO1MH101724; MvdB), a Wellcome Trust Institutional Strategic Support Fund (ISSF) award (MvdB), the Waterloo Foundation (918-1234; MvdB), the Baily Thomas Charitable Fund (2315/1; MvdB), MRC grant Intellectual Disability and Mental Health: Assessing Genomic Impact on Neurodevelopment (IMAGINE) (MR/L011166/1; JH, MvdB and MO), MRC grant Intellectual Disability and Mental Health: Assessing Genomic Impact on Neurodevelopment 2 (IMAGINE-2) (MR/T033045/1; MvdB, JH and MO); Wellcome Trust Strategic Award 'Defining Endophenotypes From Integrated Neurosciences' Wellcome Trust (100202/Z/12/Z MO, JH). NAD was supported by a National Institute for Health Research Academic Clinical Fellowship in Mental Health and MWJ by a Wellcome Trust Senior Research Fellowship in Basic Biomedical Science (202810/Z/16/Z). CE and HAM were supported by Medical Research Council Doctoral Training Grants (C.B.E. 1644194, H.A.M MR/K501347/1). HMM and UB were employed by Eli Lilly & Co during the study; HMM is currently an employee of Boehringer Ingelheim Pharma GmbH & Co KG. The views and opinions expressed are those of the author(s), and not necessarily those of the NHS, the NIHR or the Department of Health funders.