GIBH-130

Cell experiment:

The murine microglial cell line N9 (N9 cells) are cultured in DMEM supplemented with 10% FBS, 1 mM sodium pyruvate, 100 U/mL penicillin, and 100 μg/mL streptomycin. All cultures are maintained in a humidified CO2 incubator at 37°C and 5% CO2. Stock N9 cells are passaged 2-3 times/week with a 1:4 split ratio and used within 8 passages. N9 cells (5×104 cells/well) are plated into 96-well microtiter plates, followed by the treatment of Minocycline (20 μM) or various concentrations of the GIBH-130 (25, 50, and 100 μM) for 18 h. The NO production is stimulated by incubation with LPS (1 μg/mL) for 48 h. The levels of NO in the culture medium are measured using the nitric oxide detection kit[1].

Animal experiment:

Rats[1] Sprague-Dawley (SD) rats (half male and half female, 250–270 g) are used. Rats exposed to Aβ25-35 received oral (p.o.) GIBH-130 (0.0022, 0.02, or 0.18 mg/kg), Donepezil hydrochloride (0.9 mg/kg), Memantine hydrochloride (1.8 mg/kg) or distilled water via gavage after the Aβ25-35 injection. The sham-operated group receive a p.o. administration of distilled water (n=10 per group). All compounds are systemically administered in a volume of 0.01 mL per g body weight once daily after the day of surgery until the end of the behavioral testing. The behavioral tests started 8 days after the Aβ25-35 injection and are performed sequentially. Mice[1] APP/PS1 transgenic mice (half male and half female, 14 weeks age) and age-and-strained-matched wild type mice (C57BL/6J) are used. APP/PS1 transgenic mice are randomly divided into six groups: model group, GIBH-130 0.0031, 0.028, and 0.25 mg/kg group, Donepezil hydrochloride 1.3 mg/kg group, and Memantine hydrochloride 2.6 mg/kg group (n=10 per group). C57BL/6J mice are used as normal control group (n=10). The doses of GIBH-130 and the positive drugs used in this mouse model are equivalent to the rat model. All mice receive oral administration at 15 weeks of age and continuously for 25 weeks until the end of the behavioral tests. The model group and normal control group receive the same volume of distilled water (0.01 mL per g body weight).

References:

[1]. Zhou W, et al. Microglia-Based Phenotypic Screening Identifies a Novel Inhibitor of Neuroinflammation Effective in Alzheimer's Disease Models. ACS Chem Neurosci. 2016 Nov 16;7(11):1499-1507.