Kinase experiment:

Rotenone-sensitive NADH-ubiquinone oxidoreductase (complex I, CI) is assayed using 100 μM Decylubiquinone as an electron acceptor and 200 μM NADH as a donor in a 10 mM KH2PO4/K2HPO4 buffer, pH 7.5, containing 3.75 mg/mL BSA, 2 mM KCN, and 7.5 μM Antimycin A. NADH oxidation is measured at 340 nm before and after the addition of 4 μM Rotenone to allow the calculation of the Rotenone-sensitive-specific activity, which is characteristic of CI. Succinate-ubiquinone reductase (complex II, CII) activity is quantified by measuring the decrease in absorbance resulting from the reduction of 100 μM dichlorophenolindophenol at 600 nm. The measurement is performed in 50 mM KH2PO4/K2HPO4 buffer, pH 7.5, in the presence of 30 mM Succinate, 100 μM Decylubiquinone, 2 μM Rotenone, and 2 mM KCN. Coenzyme Q-cytochrome c-oxidoreductase activity (complex III, CIII), is quantified by measuring the increase in absorbance resulting from the reduction of 100 μM cytochrome c at 550 nm. The measurement is performed in 50 mM KH2PO4/K2HPO4 buffer, pH 7.5, in the presence of 100 μM Decylubiquinone previously reduced by dithionite, 50 μM EDTA, and 1 mM KCN. The specific activity is calculated by subtracting the activity obtained before and after addition of 5 μg/mL Antimycin A. 3-Hydroxyacyl-CoA dehydrogenase (HAD) activity is quantified by measuring the decrease in absorbance at 340 nm resulting from the oxidation of NADH (200 μM) and the reduction of S-acetoacetyl-CoA (50 μM). The measurement is performed in Imidazole (40 mM) and EDTA (60 μM), pH 7[1].

Animal experiment:

Mice[1]Male C57BL/6JOlaHsd mice at 4 weeks old are housed at 22℃ with a 12-h light/dark cycle. After 1 week of acclimatization, 5-6-week-old mice are divided into two groups: one with free access to a standard chow diet (SD) and the other with free access to a pelleted HFHSD diet for 16 weeks. Animals receive Imeglimin 200 mg/kg b.i.d. by oral gavage during the last 6 weeks of HFHSD feeding. Control SD and HFHSD mice are treated by oral gavage with methylcellulose 0.5% as a vehicle for drug treatment (5 mL/kg). Food intake is measured every day during the first week and twice a week until the end of the experiment. Results are expressed as grams per day per mouse.

AMP-activated protein kinase (AMPK) acts as an integrator of regulatory signals to monitor systemic and cellular energy status. Imeglimin targets on three main organs involved in glucose homeostasis: liver, muscle and the pancreas and thus has a distinct mode of action in comparison to existing Type 2 diabetes treatments.
In vitro: In liver mitochondria, imeglimin redirects substrate flows in favor of complex II. Moreover, imeglimin inhibits complex I and restores complex III activities, advicing an increase of fatty acid oxidation, further supported by an increase in hepatic 3-hydroxyacetyl-CoA dehydrogenase activity and acylcarnitine profile. Imeglimin was also found to reduce the production of reactive oxygen species and increases the DNA of mitochondrial [1].
In vivo: A previous study used a model of 16-week high-fat, high-sucrose diet (HFHSD) mice to characterize imeglimin's antidiabetic effects. Six-week imeglimin treatment could decrease glycemia, restore normal glucose tolerance, and improve insulin sensitivity, body weights and food intake significantly. This was associated with an increase of insulin-stimulated protein kinase B phosphorylation in the liver and muscle. In conclusion, imeglimin could normalize glucose tolerance and insulin sensitivity via preserving mitochondrial function from oxidative stress and favoring lipid oxidation in liver of HFHSD mice [1].
Clinical trial: Imeglimin is an experimental drug being developed as an oral anti-diabetic. Imeglimin is in development (completed phase IIb clinical trials) for use both as monotherapy and in combination with other type-2 diabetes treatments. Various research protocols are being known through various scientists. Early studies showed that imeglimin is as effective as metformin in regulating glycaemic control.
Reference:
[1] Vial G, Chauvin MA, Bendridi N, Durand A, Meugnier E, Madec AM, Bernoud-Hubac N, Pais de Barros JP, Fontaine É, Acquaviva C, Hallakou-Bozec S, Bolze S, Vidal H, Rieusset J.Imeglimin Normalizes Glucose Tolerance and Insulin Sensitivity and Improves Mitochondrial Function in Liver of a High-Fat High-Sucrose Diet Mice Model. Diabetes. 2014. pii: db141220