Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET

User menu

  • My alerts
  • Log in
  • My Cart

Search

  • Advanced search
Drug Metabolism & Disposition
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET
  • My alerts
  • Log in
  • My Cart
Drug Metabolism & Disposition

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Visit dmd on Facebook
  • Follow dmd on Twitter
  • Follow ASPET on LinkedIn
Research ArticleArticle

In Vitro Evaluation of the Disposition of A Novel Cysteine Protease Inhibitor

Wolfgang Jacobsen, Uwe Christians and Leslie Z. Benet
Drug Metabolism and Disposition November 2000, 28 (11) 1343-1351;
Wolfgang Jacobsen
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Uwe Christians
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Leslie Z. Benet
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF
Loading

Abstract

K11777 (N-methyl-piperazine-Phe-homoPhe-vinylsulfone-phenyl) is a potent, irreversible cysteine protease inhibitor. Its therapeutic targets are cruzain, a cysteine protease of the protozoan parasiteTrypanosoma cruzi, and cathepsins B and L, which are associated with cancer progression. We evaluated the metabolism of K11777 by human liver microsomes, isolated cytochrome P450 (CYP) enzymes, and flavin-containing monooxygenase 3 (FMO3) in vitro. K11777 was metabolized by human liver microsomes to three major metabolites:N-oxide K11777 (apparentKm = 14.0 ± 4.5 μM and apparentVmax = 3460 ± 3190 pmol · mg−1 · min−1, n = 4), β-hydroxy-homoPhe K11777 (Km = 16.8 ± 3.5 μM and Vmax = 1260 ± 1090 pmol · mg−1 · min−1, n = 4), andN-desmethyl K11777 (Km = 18.3 ± 7.0 μM and Vmax = 2070 ± 1830 pmol · mg−1 · min−1, n = 4). All three K11777 metabolites were formed by isolated CYP3A and their formation by human liver microsomes was inhibited by the CYP3A inhibitor cyclosporine (50 μM, 54–62% inhibition) and antibodies against human CYP3A4/5 (100 μg of antibodies/100 μg microsomal protein, 55–68% inhibition). CYP2D6 metabolized K11777 to its N-desmethyl metabolite with an apparent Km (9.2 ± 1.4 μM) lower than for CYP3A4 (25.0 ± 4.0 μM) and human liver microsomes. The apparent Km forN-oxide K11777 formation by cDNA-expressed FMO3 was 109 ± 11 μM. Based on the intrinsic formation clearances and the results of inhibition experiments (CYP2D6, 50 μM bufuralol; FMO3 mediated, 100 mM methionine) using human liver microsomes, it was estimated that CYP3A contributes to >80% of K11777 metabolite formation. K11777 was a potent (IC50 = 0.06 μM) and efficacious (maximum inhibition 85%) NADPH-dependent inhibitor of human CYP3A4 mediated 6′β-hydroxy lovastatin formation, suggesting that K11777 is not only a substrate but also a mechanism-based inhibitor of CYP3A4.

Footnotes

  • Send reprint requests to: Leslie Z. Benet, Ph.D., Professor, Department of Biopharmaceutical Sciences, School of Pharmacy, University of California, San Francisco, CA 94143-0446. E-mail: benet{at}itsa.ucsf.edu

  • This study was supported in part by National Institutes of Health Grant CA72006 (to L.Z.B), the Deutsche Forschungsgemeinschaft Grant Ch 95/6-2 (to U.C.), and National Institutes of Health NCRR Grant 01614.

  • Abbreviations used are::
    K11777
    N-methyl-piperazine-Phe-homoPhe-vinylsulfone-phenyl
    CYP
    cytochrome P450
    FMO3
    flavin-containing monooxygenase 3
    MS
    mass spectrometry
    HLF
    human liver (female)
    HLM
    human liver (male)
    LOD
    below limit of detection
    • Received November 11, 1999.
    • Accepted July 26, 2000.
  • The American Society for Pharmacology and Experimental Therapeutics
View Full Text

 

DMD articles become freely available 12 months after publication, and remain freely available for 5 years. 

Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page. 

 

  • Click here for information on institutional subscriptions.
  • Click here for information on individual ASPET membership.

 

Log in using your username and password

Forgot your user name or password?

Purchase access

You may purchase access to this article. This will require you to create an account if you don't already have one.
PreviousNext
Back to top

In this issue

Drug Metabolism and Disposition: 28 (11)
Drug Metabolism and Disposition
Vol. 28, Issue 11
1 Nov 2000
  • Table of Contents
  • About the Cover
  • Index by author
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Drug Metabolism & Disposition article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
In Vitro Evaluation of the Disposition of A Novel Cysteine Protease Inhibitor
(Your Name) has forwarded a page to you from Drug Metabolism & Disposition
(Your Name) thought you would be interested in this article in Drug Metabolism & Disposition.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Research ArticleArticle

In Vitro Evaluation of the Disposition of A Novel Cysteine Protease Inhibitor

Wolfgang Jacobsen, Uwe Christians and Leslie Z. Benet
Drug Metabolism and Disposition November 1, 2000, 28 (11) 1343-1351;

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Share
Research ArticleArticle

In Vitro Evaluation of the Disposition of A Novel Cysteine Protease Inhibitor

Wolfgang Jacobsen, Uwe Christians and Leslie Z. Benet
Drug Metabolism and Disposition November 1, 2000, 28 (11) 1343-1351;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Materials and Methods
    • Results
    • Discussion
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • Humanized PXR-CAR-CYP3A4/7 Mouse as Model of Induction
  • Ozanimod Human Metabolism and Disposition
  • High-Throughput Characterization of SLCO1B1 VUS
Show more Article

Similar Articles

Advertisement
  • Home
  • Alerts
Facebook   Twitter   LinkedIn   RSS

Navigate

  • Current Issue
  • Fast Forward by date
  • Fast Forward by section
  • Latest Articles
  • Archive
  • Search for Articles
  • Feedback
  • ASPET

More Information

  • About DMD
  • Editorial Board
  • Instructions to Authors
  • Submit a Manuscript
  • Customized Alerts
  • RSS Feeds
  • Subscriptions
  • Permissions
  • Terms & Conditions of Use

ASPET's Other Journals

  • Journal of Pharmacology and Experimental Therapeutics
  • Molecular Pharmacology
  • Pharmacological Reviews
  • Pharmacology Research & Perspectives
ISSN 1521-009X (Online)

Copyright © 2021 by the American Society for Pharmacology and Experimental Therapeutics