title: Characterization of silvestrol pharmacokinetics in mice using liquid chromatography-tandem mass spectrometry.
abstract: A sensitive and specific liquid chromatography-tandem mass spectrometry method was developed and validated for the quantification of the plant natural product silvestrol in mice, using ansamitocin P-3 as the internal standard. The method was validated in plasma with a lower limit of quantification of 1 ng/mL, accuracy ranging from 87 to 114%, and precision (coefficient of variation) below 15%. The validated method was used to characterize pharmacokinetics in C57BL/6 mice and metabolism in mouse, human and rat plasma, and liver microsomes. Mice were dosed with silvestrol formulated in hydroxypropyl-β-cyclodextrin via intravenous, intraperitoneal, and oral routes followed by blood sampling up to 24 h. Intraperitoneal systemic availability was 100%, but oral administration resulted in only 1.7% bioavailability. Gradual degradation of silvestrol was observed in mouse and human plasma, with approximately 60% of the parent drug remaining after 6 h. In rat plasma, however, silvestrol was completely converted to silvestric acid (SA) within 10 min. Evaluation in microsomes provided further evidence that the main metabolite formed was SA, which subsequently showed no cytotoxic or cytostatic activity in a silvestrol-sensitive lymphoblastic cell line. The ability of the analytical assay to measure tissue levels of silvestrol was evaluated in liver, brain, kidney, and spleen. Results indicated the method was capable of accurately measuring tissue levels of silvestrol and suggested it has a relatively low distribution to brain. Together, these data suggest an overall favorable pharmacokinetic profile of silvestrol in mice and provide crucial information for its continued development toward potential clinical testing.
authors.affiliation: College of Pharmacy, The Ohio State University, Columbus, 43210, USA.
authors.forename: U V R Vijaya
authors.initials: UV
authors.lastname: Saradhi
authors.forename: Sneha V
authors.initials: SV
authors.lastname: Gupta
authors.forename: Ming
authors.initials: M
authors.lastname: Chiu
authors.forename: Jiang
authors.initials: J
authors.lastname: Wang
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Ling
authors.forename: Zhongfa
authors.initials: Z
authors.lastname: Liu
authors.forename: David J
authors.initials: DJ
authors.lastname: Newman
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Covey
authors.forename: A Douglas
authors.initials: AD
authors.lastname: Kinghorn
authors.forename: Guido
authors.initials: G
authors.lastname: Marcucci
authors.forename: David M
authors.initials: DM
authors.lastname: Lucas
authors.forename: Michael R
authors.initials: MR
authors.lastname: Grever
authors.forename: Mitch A
authors.initials: MA
authors.lastname: Phelps
authors.forename: Kenneth K
authors.initials: KK
authors.lastname: Chan
mesh_terms: D000284:Administration, Oral; D000818:Animals; D001682:Biological Availability; D002853:Chromatography, Liquid / Q000379:methods*; D004285:Dogs; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D066298:In Vitro Techniques; D007274:Injections, Intraperitoneal; D007275:Injections, Intravenous; D057230:Limit of Detection; D051379:Mice; D008810:Mice, Inbred C57BL; D008862:Microsomes, Liver / Q000378:metabolism; D015394:Molecular Structure; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014018:Tissue Distribution; D014315:Triterpenes / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics*; D001572:Benzofurans
chemical_list: D014315:Triterpenes; C489897:silvestrol; D001572:Benzofurans