A GABA-T inhibitor
Aminooxyacetic acid is a GABA transaminase (GABA-T) inhibitor (Ki = 9.16 μM) that induces GABA accumulation in the brain.1 It also inhibits cystathionine β synthase (CBS) and cystathionine γ lyase (CSE; IC50s = 8.5 and 1.1 μM, respectively).2 Aminooxyacetic acid (100 mg/kg) decreases GABA-T activity, without affecting glutamic acid decarboxylase (GAD) activity, and increases GABA levels in all regions of rat brain.3 It also decreases GABA-T activity and increases GABA in mouse brain when administered at a dose of 13 mg/kg.4 Aminooxyacetic acid inhibits 3-mercaptopropionic acid-, strychnine-, and pentetrazole-induced seizures in mice (ED50s = 53, 130, and 85 mg/kg, respectively), however, it exhibits neurotoxicity in the chimney test (ED50 = 62 mg/kg) and is lethal (LD50 = 105 mg/kg) to mice.
1.Wallach, D.P.Studies on the GABA pathway. I. The inhibition of γ-aminobutyric acid-α-ketoglutaric acid transaminase in vitro and in vivo by U-7524 (amino-oxyacetic acid)Biochem. Pharmacol.5(4)323-331(1961) 2.Asimakopoulou, A., Panopoulos, P., Chasapis, C.T., et al.Selectivity of commonly used pharmacological inhibitors for cystathionine β synthase (CBS) and cystathionine γ lyase (CSE)Br. J. Pharmacol.169(4)922-932(2013) 3.L?scher, W., H?nack, D., and Gramer, M.Use of inhibitors of gamma-aminobutyric acid (GABA) transaminase for the estimation of GABA turnover in various brain regions of rats: A reevaluation of aminooxyacetic acidJ. Neurochem.53(6)1737-1750(1989) 4.L?scher, W.A comparative study of the pharmacology of inhibitors of GABA-metabolismNaunyn Schmiedebergs Arch. Pharmacol.315(2)119-128(1980)
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Kinase experiment: |
Enzyme activity of aspartate transaminase is measured by a colorimetric assay assessing formation of pyruvate from oxaloacetate, a product of GOT1/2 (also called AST1/2) activity, as described previously. In brief, cells grown in 6-well plates are collected after 6, 24, or 48 hours of Aminooxyacetic acid hemihydrochloride (AOA) treatment and washed with cold PBS, lysed, and supernatant used for analysis[3]. |
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Cell experiment: |
Breast cancer cell lines are used in this study. Cells are plated in 96-well plates at 1,500 to 5,000 cells per well in 100 μL media. New medium with varying concentration of Aminooxyacetic acid hemihydrochloride (AOA) is added after 12 hours. The assay is performed after 48 hours[3]. |
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Animal experiment: |
Female albino rats (150 to 200 g) bred at our department are used. Aminooxyacetic acid hemihydrochloride (AOAA) is injected into a tail vein (2 mL/kg body weight). For this purpose the rat is placed in a plastic tube and the tail warmed in water 42°C[2]. |
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参考文献: [1]. Wang C, et al. Malate-aspartate shuttle inhibitor aminooxyacetic acid leads to decreased intracellular ATP levels and altered cell cycle of C6 glioma cells by inhibiting glycolysis. Cancer Lett. 2016 Aug 1;378(1):1-7. |
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