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Comparison of the tissue distribution and metabolism of AN1284, a potent anti-inflammatory agent, after subcutaneous and oral administration in mice

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Abstract

This study is to compare the tissue distribution and metabolism of AN1284 after subcutaneous and oral administration at doses causing maximal reductions in IL-6 in plasma and tissues of mice. Anti-inflammatory activity of AN1284 and its metabolites was detected in lipopolysaccharide (LPS) activated RAW 264.7 macrophages. Mice were given AN1284 by injection or gavage, 15 min before LPS. IL-6 protein levels were measured after 4 h. Using a liquid chromatography/mass spectrometry method we developed, we showed that AN1284 is rapidly metabolized to the indole (AN1422), a 7-OH derivative (AN1280) and its glucuronide. AN1422 has weaker anti-inflammatory activity than AN1284 in LPS-activated macrophages and in mice. AN1284 (0.5 mg/kg) caused maximal reductions in IL-6 in the plasma, brain, and liver when injected subcutaneously and after gavage only in the liver. Similar reductions in the plasma and brain required a dose of 2.5 mg/kg, which resulted in 5.5-fold higher hepatic levels than after injection of 0.5 mg/kg, but 7, 11, and 19-fold lower ones in the plasma, brain, and kidneys, respectively. Hepatic concentrations produced by AN1284 were 2.5 mg/kg/day given by subcutaneously implanted mini-pumps that were only 12% of the peak levels seen after acute injection of 0.5 mg/kg. Similar hepatic concentrations were obtained by (1 mg/kg/day), administered in the drinking fluid. These were sufficient to decrease hepatocellular damage and liver triglycerides in previous experiments in diabetic mice. AN1284 can be given orally by a method of continuous release to treat chronic liver disease, and its preferential concentration in the liver should limit any adverse effects.

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Acknowledgements

We thank Donna Schorer-Apelbaum for performing the graphics and statistical analyses and Yulia Shenberger and Bruria Schmerling for the help with the LC-MS/MS analysis.

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Authors and Affiliations

  1. Department of Chemistry, Bar-Ilan University, 5290002, Ramat Gan, Israel

    Michal Weitman, Inessa Yanovsky, Shani Zeeli & Abraham Nudelman

  2. Institute of Drug Research, School of Pharmacy, The Hebrew University of Jerusalem, Ein Kerem, 9112002, Jerusalem, Israel

    Corina Bejar, Michal Melamed, Tehilla Weill & Marta Weinstock

Authors
  1. Michal Weitman
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  2. Corina Bejar
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  3. Michal Melamed
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  4. Tehilla Weill
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  5. Inessa Yanovsky
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  6. Shani Zeeli
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  7. Abraham Nudelman
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  8. Marta Weinstock
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Contributions

All authors read and approved the manuscript and all data were generated in-house.

MaW: Conceptualization; supervision, calculations, writing.

CB, MM, TW: all biological experimentation.

IY, SZ: chemical syntheses.

MiW: Analytical and mass spectral studies.

AN: Supervision of spectral analyses and chemical syntheses.

Corresponding author

Correspondence to Marta Weinstock.

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All experiments were performed according to the ethical committee guidelines of The Hebrew University Medical School, numbers MD-18–15512-4 and MD-20–16205-2.

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The authors declare no competing interests.

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Weitman, M., Bejar, C., Melamed, M. et al. Comparison of the tissue distribution and metabolism of AN1284, a potent anti-inflammatory agent, after subcutaneous and oral administration in mice. Naunyn-Schmiedeberg's Arch Pharmacol 394, 2077–2089 (2021). https://doi.org/10.1007/s00210-021-02125-y

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