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de Angelis, M. ; Schriever, S.C. ; Kyriakou, E. ; Sattler, M. ; Messias, A.C. ; Schramm, K.-W. ; Pfluger, P.T.

Detection and quantification of the anti-obesity drug celastrol in murine liver and brain.

Neurochem. Int. 136:104713 (2020)

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Celastrol is a natural pentacyclic triterpene extracted from the roots of Tripterygium wilfordi (thunder god vine). Celastrol was reported as a powerful anti-obesity drug with leptin sensitizing properties that decreases food consumption and mediates body weight loss when administered to diet-induced obese mice at 100 mu g/kg body weight. The weight lowering properties of celastrol are likely mediated by the CNS, in particular, by the hypothalamus, but the final proof for the accumulation of celastrol in the brain and hypothalamus remains to be established. Here, we aimed to demonstrate that intraperitoneal celastrol administration at 100 mu g/kg can rapidly reach the brain and, in particular, the hypothalamus of mice. We developed and validated a sensitive liquid chromatography mass spectrometry method for the quantitative determination of celastrol in murine tissues, namely liver, brain and hypothalamus. Chow-fed lean mice were randomly assigned to the vehicle vs. celastrol groups, injected with saline or 100 mu g/kg body weight of celastrol, and sacrificed 30 min or 120 min post injection. Celastrol was extracted from homogenized tissue using ethyl acetate as organic solvent, and quantified using a matrix-matched calibration curve with glycyrrhetinic acid as internal standard. Liver celastrol concentrations were 32.60 +/- 8.21 pg/mg and 40.52 +/- 15.6 pg/mg, 30 and 120 min after injection, respectively. We found 4.70 +/- 0.31 pg/mg celastrol after 30 min, and 16.22 +/- 3.33 pg/mg after 120 min in whole brain lysates, and detectable amounts in the hypothalamus. These results corroborate the validity of our methodology, demonstrate the accumulation of celastrol in the brain of mice injected intraperitoneally with a dose of 100 mu g/ kg, and confirm the CNS as possible site of action for the weight lowering properties of celastrol.

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