DMD Large equally mixed donor pool

Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
QUICK SEARCH:   [advanced]


     

Drug Metabolism and Disposition Fast Forward
First published on May 31, 2007; DOI: 10.1124/dmd.107.014746

0090-9556/07/3509-1554-1563$20.00
DMD 35:1554-1563, 2007

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Erratum
Right arrow An erratum has been published
Right arrow All Versions of this Article:
dmd.107.014746v1
35/9/1554    most recent
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Cai, P.
Right arrow Articles by Ruppen, M. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Cai, P.
Right arrow Articles by Ruppen, M. E.

In Vitro Metabolic Study of Temsirolimus: Preparation, Isolation, and Identification of the Metabolites

Ping Cai, Rushung Tsao, and Mark E. Ruppen

Chemical and Pharmaceutical Development, Wyeth Research, Pearl River, New York

The in vitro metabolism of temsirolimus, (rapamycin-42-[2,2-bis-(hydroxymethyl)]-propionate), an antineoplastic agent, was studied using human liver microsomes as well as recombinant human cytochrome P450s, namely CYP3A4, 1A2, 2A6, 2C8, 2C9, 2C19, and 2E1. Fifteen metabolites were detected by liquid chromatography (LC)-tandem mass spectrometry (MS/MS or MS/MS/MS). CYP3A4 was identified as the main enzyme responsible for the metabolism of the compound. Incubation of temsirolimus with recombinant CYP3A4 produced most of the metabolites detected from incubation with human liver microsomes, which was used for large-scale preparation of the metabolites. By silica gel chromatography followed by semipreparative reverse-phase high-performance liquid chromatography, individual metabolites were separated and purified for structural elucidation and bioactivity studies. The minor metabolites (peaks 1-7) were identified as hydroxylated or desmethylated macrolide ring-opened temsirolimus derivatives by both positive and negative mass spectrometry (MS) and MS/MS spectroscopic methods. Because these compounds were unstable and only present in trace amounts, no further investigations were conducted. Six major metabolites were identified as 36-hydroxyl temsirolimus (M8), 35-hydroxyl temsirolimus (M9), 11-hydroxyl temsirolimus with an opened hemiketal ring (M10 and M11), N- oxide temsirolimus (M12), and 32-O-desmethyl temsirolimus (M13) using combined LC-MS, MS/MS, MS/MS/MS, and NMR techniques. Compared with the parent compound, these metabolites showed dramatically decreased activity against LNCaP cellular proliferation.

Address correspondence to: Dr. Ping Cai, Wyeth Research, Building 250/229, 401 N. Middletown Rd., Pearl River, NY 10965. E-mail: caip{at}wyeth.com




This article has been cited by other articles:


Home page
 Clin. Cancer Res.Home page
J. G. Kuhn, S. M. Chang, P. Y. Wen, T. F. Cloughesy, H. Greenberg, D. Schiff, C. Conrad, K. L. Fink, H. I. Robins, M. Mehta, et al.
Pharmacokinetic and Tumor Distribution Characteristics of Temsirolimus in Patients with Recurrent Malignant Glioma
Clin. Cancer Res., December 15, 2007; 13(24): 7401 - 7406.
[Abstract] [Full Text] [PDF]


Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
All ASPET Journals Molecular Pharmacology Pharmacological Reviews
Molecular Interventions Drug Metabolism and Disposition

Copyright © 2007 by the American Society for Pharmacology and Experimental Therapeutics.