Abstract 4879: Determination of chebulagic acid and chebulinic acid and their pharmacokinetics in mouse .

Purpose: Triphala (THL), an ayurveda powder mixture of three myrobalan fruits, has shown promising anti-tumor growth activity in stomach and pancreatic tumor mouse models. Chebulagic acid (CA) and chebulinic acid (CI) are two bioactive THL tannins showing preventive effect on diabetes and carcinogenesis. Notably, our study demonstrated that THL and CI can inhibit vascular endothelial growth factor (VEGF) induced angiogenesis, a critical step for the initiation, growth, progression of malignant tumor. However, the pharmacokinetics (PK) of CA and CI remains unknown. Therefore, the purpose of this study was to establish an LC-MS/MS method to measure CA and CI in mouse plasma and to characterize their PK in mice after an oral administration of THL. Methods: The following ion transitions at m/zs: 953.1/303.0, 955.3/337.2, and 301.5/163.9 were used for monitoring CA, CI and hesperetin (internal standard), respectively, on an API-3000 mass spectrometer. THL (100 mg/kg) in water containing 22.2 mg/kg CA and 6.8 mg/kg CI was orally gavaged to mice. Plasma samples were collected at the following time points, 10, 20, 30, 60, 120, 240 and 480 min after dose. The plasma level of CA and CI was determined using a LC-MS/MS method. The PK parameters of CA and CI were retrieved using a Computer Software WinNonlin (Pharasight version 5.3). Results: An LC-MS/MS method for determination of CA and CI was developed and validated in the concentration range of 1-1000 ng/mL with the low limit quantification of 5 ng/mL in mouse plasma. This method was capable to characterize the pharmacokinetics of CA and CI in mouse after an oral administration of 100 mg/kg THL. The plasma concentration and time profile of CA and CI demonstrated that both CA and CI was absorbed rapidly with an absorption half-life (K01-HL) of 1.19 min and 6.71 min, respectively. The peak plasma concentration (Cmax) of CA and CI reached 4061 (∼4.0 μM) and 1953 ng/mL (∼2.0 μM) in plasma. After absorption, CA and CI eliminate rapidly with a half-life (K10-HL) of 67 min and 40.5 min, respectively. Both are detectable up to 8 h in mouse plasma. The area under the plasma concentration-time curve (AUC) to the last time point (480 min) of CA and CI are 4.24E+05 min*ng/ml and 1.23E+05 ng/ml.min and their volume distribution (V_F) are 5083 mL/kg and 3235 mL/kg and their clearance (Cl-F) is 55.5 mL/min/kg and 52.6 mL/min/kg, respectively. Conclusions: The pharmacological effective concentration of CI targeting VEGF (2.0 μM) can be achieved in plasma in mice after an oral administration of non-toxic THL dose. The favorable pharmacokinetic characteristics of CI along with the LC-MS/MS method warrants further exploration of the potential correlation of pharmacokinetics and pharmacodynamics of CI or THL as a chemopreventive and chemotherapeutic agent targeting VEGF. Supported by biomedical mass spectrometric laboratory Citation Format: Di Bei, Leon Xie, Kai Lu, Kenneth Chan, Sujit Basu, Zhongfa Liu. Determination of chebulagic acid and chebulinic acid and their pharmacokinetics in mouse . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4879. doi:10.1158/1538-7445.AM2013-4879