Metformin hydrochloride c4h12cin5

Due to its generic use in the clinical management of individuals with PCOS and to generate a syngeneic control cohort comparing the therapeutic efficacy of tacrolimus in a murine model of PCOS, the broadly prescribed antidiabetic medication metformin was used in the HFD-dNONcNZO mice (Supplemental Table S1 and Fig. S1). Metformin Hydrochloride (C4H₁₂CIN₅, Catalogue #M258815, Toronto Research Chemicals, Toronto, ON) was prepared fresh daily and made available to the HFD-dNONcNZO mice ad libitum (200 mg/dL) per day from age weeks 15 to 20 according to an established protocol^{28 (link)}.

Due to the low bioavailability of tacrolimus (14–32%) [16 (link)] and data obtained in several pilot attempts at determining the optimum dosage of tacrolimus for use in this particular study, a modified tacrolimus-based monotherapy protocol was developed for use in the HFD-dNONcNZO mice. Tacrolimus (FK506) (Tacrolimus-Astellas Pharma US, Inc., Deerfield, IL) was dissolved in castor oil and ethanol (vehicle for FK506 and untreated groups) and administered to the diabetic HFD-dNONcNZO mice on an alternate day regimen from week 16–19 and week 25–29, respectively (Additional file 1: Figure S1, Table S1). For proper comparisons and adequacy in providing a standard diabetic drug-treated T2DM diabetic mouse cohort for this study, metformin (Metformin Hydrochloride (C4H12CIN5), Catalogue # M258815, Toronto Research Chemicals, Toronto, ON) was selected for use in HFD-dNONcNZO mice to generate genetically matched treated control. Due to the significant first pass effect, metformin doses in diabetic rodents are typically ≥50 mg/day [17 (link)]. Using this as a starting point and knowing that average water consumption in the diabetic mouse is ~10–15× the upper limit of normoglycemic mouse water consumption of 7.7 + 0.3 mL/30 g bwt [18 (link)], metformin was prepared fresh daily and made available ad libitum (200 mg/dL) per day.