Glucophage

The metformin drug used in this trial is Glucophage, a commercially available extended release metformin-hydrochloride pill, owned by Merck GmbH, Vienna, Austria. It is packaged in boxes containing 200 pills with no special markings regarding the trial.

High-dose cyclophosphamide (Endoxan®, Baxter, CA^®) was administered at a dose of 100–150 mg/kg by intraperitoneal injection every other day for 3 weeks followed by 2 weeks of rest.
Metronomic cyclophosphamide was described previously [23 (link)]. The drug was dissolved in distilled water (stock solution 20 mg/ml) and kept at 4 °C in the dark for one week. A total of 1.25 ml of stock solution was dissolved in 200 ml water at a concentration of 20 mg/kg dose.
The metformin “Glucophage^®, Merck, NJ” dosage was 5 mg/ml dissolved in drinking water to provide a dose of 5 mg/kg. Drinking water was changed twice weekly.

The patients were examined by the gynecologists. The subjects were randomly selected to receive either MTF and NAC or placebo. Eighty patients were divided into four groups (N=20): i. NAC (NAC group) which received NAC (Holzkrichen, Germany, batch no. 6N5483; 600 mg three times daily), ii. MTF group which received MTF (Glucophage, Merck, West Drayton, UK; 500 mg three times daily), iii. MTF+NAC group which were co-treated with MTF and NAC with the offered doses three times daily, and iv. placebo (PLA group) which received oral rehydration salts (ORS, Poursina, Tehran, Iran; batch no.30) three times daily, for a period of six weeks. Before beginning treatment, patients were also asked to report any possible adverse effects during the treatment, and they were evaluated for any presenting main complaints at the end of the treatment period.
A total number of 20 patients (6 from MTF group, 4 from NAC group, 6 from MTF+NAC group and 4 from PLA group) were dropped out due to intolerance of medication, monofollicular development, and failure of ovulation induction during ICSI cycle, and ultimately, 60 patients remained in the study.

Overnight fasted mice were treated either with saline (vehicle) or metformin (Glucophage; Merck, France) at 400 mg/kg of body weight administered by oral gavage. After 30 min animals were given 75 mg of D-glucose by injecting into the lumen of the duodenum of mice anesthetized with isoflurane. Ten min later, concentration of glucose was measured in blood sample collected from portal vein.

Overnight fasted mice were treated either with saline (vehicle) or M400, M200 and M60, respectively(Glucophage; Merck, France) administered by oral gavage. Thirty min later animals were given D-glucose either orally at a dose 3 g/kg or intraperitoneally at a dose 1 g/kg and subjected to GTT. The degree of glucose intolerance was quantified as AUC. Insulin levels were measured in plasma obtained from blood samples collected before and 30 min after glucose administration, using the Sensitive rat insulin RIA kit (Merck Millipore, USA). Compound C (20 mg/kg body weight; Abcam), Exendin 9–39 (0.1 mg/kg body weight; Sigma Aldrich) and MK-801 (0.3 mg/kg body weight; Sigma Aldrich), i.e. the pharmaceutical antagonists of the AMPK, GLP1R and NMDAR, respectively, were administered via i.p. injections 30 min (Compound C) or 10 min (Exendin 9–39, MK-801) before metformin.