Forxiga

The mice were administered CANA (Invocana, Janssen Pharmaceuticals, Belgium) or DAPA (Forxiga, Astra Zeneca, UK) using an oral gastric tube, once daily at doses of 100 and 10 mg/kg, respectively, in a volume of 3 ml/kg in 5% water solution of gum arabic for 14 days (from 14 to 28th day of the experiment). The STZ groups were administered CANA and DAPA 14 days after the STZ injection. Doses were selected based on the pharmacokinetics of CANA and DAPA in mice^{18 (link)}. The Contr Norm and Contr STZ groups received an equal volume of a 5% water solution of gum arabic (from 14 to 28th day of the experiment).

The DAPA propanediol monohydrate raw material was obtained from Lianchuang Biological Pharmaceutical Co. (Anhui, China, Batch: 20150207). Five lots of Forxiga (AstraZeneca, Cotia, SP, Brazil) tablets, two lots containing 5 mg of DAPA (identified in this study as lots A and B) and three lots containing 10 mg of DAPA (C, D, and E), were bought in commercial pharmacies.

Male, 8-week-old, C57BL/6J littermate mice were purchased from Damul Science (Daejeon, Korea), housed in a temperature-controlled facility (22 ± 1 °C) that maintained a 12 h light/dark cycle, and provided free access to a standard chow diet (5L79, LabDiet, St. Louis, MO, USA) and water. A schematic representation of the study design is shown in Figure 1A. Mice were randomly assigned to each study group. To prepare the acute heart failure rodent model, mice were administered a single dose of 15 mg/kg doxorubicin (Sigma-Aldrich, Oakville, ON, Canada) or 0.9% sterile saline. To prepare the chronic heart failure model, serial intraperitoneal injections of doxorubicin (2.5 mg/kg) or 0.9% sterile saline were administered every 4 days for 24 days (cumulative dosage, 15 mg/kg). One day after doxorubicin injection, the control mice were gavaged daily with vehicle (corn oil) (Sigma-Aldrich), while the experimental mice were gavaged daily with ARNI (ENTRESTO^®; Novartis, Basel, Switzerland) (68 mg/kg/day), SGLT2 inhibitor (Forxiga^®; AstraZeneca, Cambridge, UK) (1 mg/kg/day), Low-ARNI/SGLT2i (Low-ARNI, 34 mg/kg/day; SGLT2i, 1 mg/kg/day), and ARNI/SGLT2i for 6 weeks. Dosage of ARNI and SGLT2i for our study was based on prior preclinical studies [22 (link),23 (link)]. For the collection of blood and tissue samples, animals were euthanized by isoflurane overdose followed by exsanguination.