title: A local effect of CYP24 inhibition on lung tumor xenograft exposure to 1,25-dihydroxyvitamin D(3) is revealed using a novel LC-MS/MS assay.
abstract: The vitamin D(3) catabolizing enzyme, CYP24, is frequently over-expressed in tumors, where it may support proliferation by eliminating the growth suppressive effects of 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)). However, the impact of CYP24 expression in tumors or consequence of CYP24 inhibition on tumor levels of 1,25(OH)(2)D(3)in vivo has not been studied due to the lack of a suitable quantitative method. To address this need, an LC-MS/MS assay that permits absolute quantitation of 1,25(OH)(2)D(3) in plasma and tumor was developed. We applied this assay to the H292 lung tumor xenograft model: H292 cells eliminate 1,25(OH)(2)D(3) by a CYP24-dependent process in vitro, and 1,25(OH)(2)D(3) rapidly induces CYP24 expression in H292 cells in vivo. In tumor-bearing mice, plasma and tumor concentrations of 1,25(OH)(2)D(3) reached a maximum of 21.6 and 1.70ng/mL, respectively, following intraperitoneal dosing (20μg/kg 1,25(OH)(2)D(3)). When co-administered with the CYP24 selective inhibitor CTA091 (250μg/kg), 1,25(OH)(2)D(3) plasma levels increased 1.6-fold, and tumor levels increased 2.6-fold. The tumor/plasma ratio of 1,25(OH)(2)D(3) AUC was increased 1.7-fold by CTA091, suggesting that the inhibitor increased the tumor concentrations of 1,25(OH)(2)D(3) independent of its effects on plasma disposition. Compartmental modeling of 1,25(OH)(2)D(3) concentration versus time data confirmed that: 1,25(OH)(2)D(3) was eliminated from plasma and tumor; CTA091 reduced the elimination from both compartments; and that the effect of CTA091 on tumor exposure was greater than its effect on plasma. These results provide evidence that CYP24-expressing lung tumors eliminate 1,25(OH)(2)D(3) by a CYP24-dependent process in vivo and that CTA091 administration represents a feasible approach to increase tumor exposure to 1,25(OH)(2)D(3).
authors.affiliation: Molecular Therapeutics/Drug Discovery Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, United States.
authors.forename: Jan H
authors.initials: JH
authors.lastname: Beumer
authors.forename: Robert A
authors.initials: RA
authors.lastname: Parise
authors.forename: Beatriz
authors.initials: B
authors.lastname: Kanterewicz
authors.forename: Martin
authors.initials: M
authors.lastname: Petkovich
authors.forename: David Z
authors.initials: DZ
authors.lastname: D'Argenio
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Hershberger
mesh_terms: D000818:Animals; D019540:Area Under Curve; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005240:Feasibility Studies; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008819:Mice, Nude; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013250:Steroid Hydroxylases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D014807:Vitamin D / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D065668:Vitamin D3 24-Hydroxylase; D014815:Vitamins / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics; D023041:Xenograft Model Antitumor Assays
chemical_list: D004791:Enzyme Inhibitors; D014815:Vitamins; D014807:Vitamin D; C097949:1,25-dihydroxyvitamin D; D013250:Steroid Hydroxylases; D065668:Vitamin D3 24-Hydroxylase