Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Special Sections
    • 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
  • Submit
  • 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
    • Special Sections
    • 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
  • Submit
  • Visit dmd on Facebook
  • Follow dmd on Twitter
  • Follow ASPET on LinkedIn
Research ArticleArticle

Commensal Gut Bacteria Convert the Immunosuppressant Tacrolimus to Less Potent Metabolites

Yukuang Guo, Camila Manoel Crnkovic, Kyoung-Jae Won, Xiaotong Yang, John Richard Lee, Jimmy Orjala, Hyunwoo Lee and Hyunyoung Jeong
Drug Metabolism and Disposition March 2019, 47 (3) 194-202; DOI: https://doi.org/10.1124/dmd.118.084772
Yukuang Guo
Departments of Medicinal Chemistry and Pharmacognosy (Y.G., C.M.C., J.O., H.L.), Pharmacy Practice (K.-J.W., H.J.), Biopharmaceutical Sciences (X.Y., H.J.), and Center for Biomolecular Sciences (Y.G., J.O., H.L., H.J.), University of Illinois at Chicago, Chicago, Illinois; and Division of Nephrology and Hypertension, Department of Medicine, Weill Cornell Medicine, New York, New York (J.R.L.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Camila Manoel Crnkovic
Departments of Medicinal Chemistry and Pharmacognosy (Y.G., C.M.C., J.O., H.L.), Pharmacy Practice (K.-J.W., H.J.), Biopharmaceutical Sciences (X.Y., H.J.), and Center for Biomolecular Sciences (Y.G., J.O., H.L., H.J.), University of Illinois at Chicago, Chicago, Illinois; and Division of Nephrology and Hypertension, Department of Medicine, Weill Cornell Medicine, New York, New York (J.R.L.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kyoung-Jae Won
Departments of Medicinal Chemistry and Pharmacognosy (Y.G., C.M.C., J.O., H.L.), Pharmacy Practice (K.-J.W., H.J.), Biopharmaceutical Sciences (X.Y., H.J.), and Center for Biomolecular Sciences (Y.G., J.O., H.L., H.J.), University of Illinois at Chicago, Chicago, Illinois; and Division of Nephrology and Hypertension, Department of Medicine, Weill Cornell Medicine, New York, New York (J.R.L.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Xiaotong Yang
Departments of Medicinal Chemistry and Pharmacognosy (Y.G., C.M.C., J.O., H.L.), Pharmacy Practice (K.-J.W., H.J.), Biopharmaceutical Sciences (X.Y., H.J.), and Center for Biomolecular Sciences (Y.G., J.O., H.L., H.J.), University of Illinois at Chicago, Chicago, Illinois; and Division of Nephrology and Hypertension, Department of Medicine, Weill Cornell Medicine, New York, New York (J.R.L.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
John Richard Lee
Departments of Medicinal Chemistry and Pharmacognosy (Y.G., C.M.C., J.O., H.L.), Pharmacy Practice (K.-J.W., H.J.), Biopharmaceutical Sciences (X.Y., H.J.), and Center for Biomolecular Sciences (Y.G., J.O., H.L., H.J.), University of Illinois at Chicago, Chicago, Illinois; and Division of Nephrology and Hypertension, Department of Medicine, Weill Cornell Medicine, New York, New York (J.R.L.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jimmy Orjala
Departments of Medicinal Chemistry and Pharmacognosy (Y.G., C.M.C., J.O., H.L.), Pharmacy Practice (K.-J.W., H.J.), Biopharmaceutical Sciences (X.Y., H.J.), and Center for Biomolecular Sciences (Y.G., J.O., H.L., H.J.), University of Illinois at Chicago, Chicago, Illinois; and Division of Nephrology and Hypertension, Department of Medicine, Weill Cornell Medicine, New York, New York (J.R.L.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hyunwoo Lee
Departments of Medicinal Chemistry and Pharmacognosy (Y.G., C.M.C., J.O., H.L.), Pharmacy Practice (K.-J.W., H.J.), Biopharmaceutical Sciences (X.Y., H.J.), and Center for Biomolecular Sciences (Y.G., J.O., H.L., H.J.), University of Illinois at Chicago, Chicago, Illinois; and Division of Nephrology and Hypertension, Department of Medicine, Weill Cornell Medicine, New York, New York (J.R.L.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hyunyoung Jeong
Departments of Medicinal Chemistry and Pharmacognosy (Y.G., C.M.C., J.O., H.L.), Pharmacy Practice (K.-J.W., H.J.), Biopharmaceutical Sciences (X.Y., H.J.), and Center for Biomolecular Sciences (Y.G., J.O., H.L., H.J.), University of Illinois at Chicago, Chicago, Illinois; and Division of Nephrology and Hypertension, Department of Medicine, Weill Cornell Medicine, New York, New York (J.R.L.)
  • 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 + SI
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • Additional Files
  • Fig. 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 1.

    F. prausnitzii metabolizes tacrolimus. (A) F. prausnitzii (OD600 2.6) cultured in YCFA media was incubated with tacrolimus (100 μg/ml) anaerobically at 37°C for 48 hours. The mixture was analyzed by using HPLC/UV. (B) Time profiles of tacrolimus disappearance and M1 appearance upon anaerobic incubation of tacrolimus (100 μg/ml) with F. prausnitzii.

  • Fig. 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    Chemical structures of tacrolimus and F. prausnitzii–derived metabolite M1. M1 structure was identified using mass spectrometry and nuclear magnetic resonance spectroscopy.

  • Fig. 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 3.

    M1 is less potent than tacrolimus as an immunosuppressant and antifungal agent. (A) Immunosuppressant activities of tacrolimus and M1 were examined in PBMCs by measuring cell proliferation after treatment with a T-lymphocyte mitogen in the presence of tacrolimus or M1. (B) Antifungal activities of tacrolimus and M1 were examined using Malassezia sympodialis. The yeast was inoculated on a modified Dixon agar plate. After 1 hour incubation, an aliquot of tacrolimus or M1 at different concentrations was placed on the plate, as shown in the left panel, and incubated at 37°C for 2 days.

  • Fig. 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 4.

    Multiple commensal gut bacteria convert tacrolimus to M1. (A) Representative chromatograms of bacteria incubated with tacrolimus. M1 nonproducer (Bifidobacterium longum) or producer (Clostridium aldenense, Clostridium citroniae, and Erysipelotrichaceae sp.) cultured overnight in YCFA media was incubated with tacrolimus (100 μg/ml) anaerobically at 37°C for 48 hours. The mixture was analyzed by using HPLC/UV at 210 nm. (B) Mouse or human hepatic microsomes [(HMs); 3 mg microsomal protein/ml] were incubated with tacrolimus (100 μg/ml) at 37°C for 2 hours aerobically. The mixture was analyzed by using HPLC/UV.

  • Fig. 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 5.

    Human gut microbiota convert tacrolimus to M1. Tacrolimus (100 μg/ml) was incubated anaerobically with human stool samples from two different subjects (100 mg wet weight/ml) for 48 hours at 37°C. A separate set of samples was boiled for 10 minutes before incubation with tacrolimus. The incubation mixtures were analyzed by HPLC/UV.

  • Fig. 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 6.

    M1 formation by small intestinal bacteria may be extensive. (A) Tacrolimus (10 μg/ml) was incubated anaerobically with F. prausnitzii (6.3 × 107 cells/ml) for varying amounts of time. (B) Tacrolimus (10 μg/ml) was incubated with varying amount of F. parusnitzii in PBS at 37°C for 2 hours. (C) Tacrolimus at varying concentrations was incubated with F. prausnitzii (6.3 × 107 cells/ml) for 1 hour. M1 concentrations in the reaction mixtures were analyzed by liquid chromatography–MS/MS.

Tables

  • Figures
  • Additional Files
    • View popup
    TABLE 1

    Screening gut bacteria for tacrolimus conversion to M1 in YCFA culture

    OrderBacteriumOD600M1 Production Detected
    BifidobacterialesBifidobacterium longum1.8No
    BacteroidalesBacteroides cellulosilyticus0.6Yesa
    Bacteroides finegoldii3.4Yesa
    Bacteroides ovatus4.2Yesa
    Parabacteroides merdae2.7Yesa
    Parabacteroides johnsonii3.6Yesa
    Parabacteroides goldsteinii3.3Yesa
    ClostridialesRuminococcaceae sp.0.5Yes
    Clostridium innocuum3.4Yes
    Anaerostipes sp.2.7Yes
    Dorea formicigenerans2.4Yes
    Clostridium clostridioforme3.0Yes
    Clostridium hathewayi2.6Yes
    Blautia sp.4.7Yes
    Clostridium aldenense1.4Yes
    Clostridium symbiosum2.5Yes
    Clostridium citroniae1.7Yes
    Coprococcus sp.2.4Yes
    Clostridium bolteae3.6Yes
    Clostridium cadaveris1.4Yes
    Ruminococcus gnavus3.4Yes
    ErysipelotrichalesErysipelotrichaceae sp.3.8Yes
    • ↵a M1 production observed only when using sensitive HPLC-MS/MS for detection.

    • View popup
    TABLE 2

    M1 levels in kidney transplant patients’ stool samples

    PatientAgeGenderPost-Transplant DayTacrolimus Oral DoseaFecal AbundanceBaseline Level in Stool SamplesM1 Production upon Tacrolimus Incubation
    F. prausnitziiClostridialesTacrolimusM1
    yrmg/day%%ng/mg stoolng/mg stoolng/mg stool
    145Female31946860.880.385.1
    256Male1833989BQLbBQLb3.5
    361Male20532710.63BQLb4.5
    459Female12627760.710.122.9
    550Male321026790.370.416.4
    652Female286ND150.29BQLb3.5
    757Male153ND440.85BQLb4.1
    871Male184ND95BQLb0.607.1
    925Male274ND740.49BQLb12.6
    1052Male326ND950.14BQLb11.0
    • BQL, below the quantification limit; ND, not detected.

    • ↵a At the time of stool collection.

    • ↵b Below the quantification limit (i.e., 0.1 ng/mg stool).

Additional Files

  • Figures
  • Tables
  • Data Supplement

    • Supplemental Data -

      Supplemental Figure 1 - Different strains of F. prausnitzii (i.e., A2-165, ATCC 27766 and ATCC 27768) were cultured overnight in YCFA media, followed by anaerobic incubation with tacrolimus (100 μg/ml) at 37°C for 48 h

      Supplemental Figure 2 - MS2 spectra of tacrolimus and M1 (IT-TOF)

      Supplemental Figure 3 - MS2 fragments of tacrolimus and M1 obtained in IT-TOF and QTOF systems

      Supplemental Figure 4 - Tacrolimus (50 mg) was incubated with F. prausnitzii A2-165 (OD ~2; 500 ml) for 96 h, followed by ethyl acetate extraction of the mixture

      Supplemental Figure 5 - IR spectra of a) tacrolimus, b) M1, c) overlay of tacrolimus (red) and M1 (blue) spectra

      Supplemental Figure 6 - 1H NMR spectrum (900 MHz, CDCl3) of M1

      Supplemental Figure 7 - DEPTQ spectrum (226 MHz, CDCl3) of M1

      Supplemental Figure 8 - COSY spectrum (900 MHz, CDCl3) of M1

      Supplemental Figure 9 - HSQC spectrum (900 MHz, CDCl3) of M1

      Supplemental Figure 10 - HMBC spectrum (900 MHz, CDCl3) of M1

      Supplemental Figure 11 - 1H-13C HSQC-TOCSY spectrum (900 MHz, CDCl3, 90 ms mixing time) of M1

      Supplemental Figure 12 - Key 2D NMR correlations of M1 (Isomer I)

      Supplemental Figure 13 - Expansions of the DEPTQ spectra (226 MHz, CDCl3) of M1 (three major isomers assigned) and tacrolimus (two isomers)

      Supplemental Figure 14 - Bacteria cultured overnight in YCFA media were incubated with tacrolimus (100μg/ml) or vehicle control anaerobically at 37°C for 48 h

      Supplemental Figure 15 - Tacrolimus (10 μg/ml) was incubated with F. prausnitzii (6.28×107 CFU/ml) anaerobically at 37°C up to 24 h

      Supplemental Table 1 -  List of commensal gut bacteria obtained from Biodefense and Emerging Infections (BEI) Research Resources Repository

      Supplemental Table 2 - Demographic information of kidney transplant patients whose stool samples were analyzed

      Supplemental Table 3 - NMR spectroscopic data of M1 (Isomer I), CDCl3a

      Supplemental Table 4 - NMR Spectroscopic Data of M1 (Isomer II), CDCl3a

PreviousNext
Back to top

In this issue

Drug Metabolism and Disposition: 47 (3)
Drug Metabolism and Disposition
Vol. 47, Issue 3
1 Mar 2019
  • Table of Contents
  • Table of Contents (PDF)
  • About the Cover
  • Index by author
  • Editorial Board (PDF)
  • Front Matter (PDF)
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.
Commensal Gut Bacteria Convert the Immunosuppressant Tacrolimus to Less Potent Metabolites
(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

Tacrolimus Metabolism by Gut Bacteria

Yukuang Guo, Camila Manoel Crnkovic, Kyoung-Jae Won, Xiaotong Yang, John Richard Lee, Jimmy Orjala, Hyunwoo Lee and Hyunyoung Jeong
Drug Metabolism and Disposition March 1, 2019, 47 (3) 194-202; DOI: https://doi.org/10.1124/dmd.118.084772

Citation Manager Formats

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

Share
Research ArticleArticle

Tacrolimus Metabolism by Gut Bacteria

Yukuang Guo, Camila Manoel Crnkovic, Kyoung-Jae Won, Xiaotong Yang, John Richard Lee, Jimmy Orjala, Hyunwoo Lee and Hyunyoung Jeong
Drug Metabolism and Disposition March 1, 2019, 47 (3) 194-202; DOI: https://doi.org/10.1124/dmd.118.084772
del.icio.us logo Digg logo Reddit logo Twitter logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Introduction
    • Materials and Methods
    • Results
    • Discussion
    • Acknowledgments
    • Authorship Contributions
    • Footnotes
    • Abbreviations
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF + SI
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • CES2-based hydrolysis of molnupiravir
  • Oral PBPK Modeling of Vismodegib
  • Transporter-enzyme interplay in PK of PF-06835919
Show more Articles

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 © 2022 by the American Society for Pharmacology and Experimental Therapeutics