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Absorption and metabolism of genistein and its five isoflavone analogs in the human intestinal Caco-2 model

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Abstract

The purposes of this study were to determine the effect of structural change on the intestinal disposition of isoflavones and to elucidate the mechanisms responsible for transport of phase II isoflavone conjugates. Transport and metabolism of six isoflavones (i.e., genistein, daidzein, glycitein, formononetin, biochanin A, and prunetin) were studied in the human intestinal Caco-2 model and mature Caco-2 cell lysate. Glucuronides were the main metabolites in intact Caco-2 cells for all isoflavones except prunetin, which was mainly sulfated. In addition, the 7-hydroxy group was the main site for glucuronidation whereas the 4′-hydroxy group was only one of the possible sites for sulfation. Glucuronidated isoflavones (except biochanin A) were preferably excreted to the basolateral side, whereas sulfated metabolites (except genistein and glycitein) were mainly excreted into the apical side. Polarized excretion of most isoflavone conjugates was inhibited by the multidrug resistance-related protein (MRP) inhibitor leukotriene C4 (0.1 μM) and the organic anion transporter (OAT) inhibitor estrone sulfate (10 μM). When formation and excretion rates of isoflavones were determined simultaneously, the results showed that formation served as the rate-limiting step for all isoflavone conjugates (both glucuronides and sulfates) except for genistein glucuronide, which had comparable excretion and formation rates. In conclusion, the intestinal disposition of isoflavones was structurally dependent, polarized, and mediated by MRP and OAT. Formation generally served as the rate-limiting step in the cellular excretion of conjugated isoflavones in the Caco-2 cell culture model.

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Abbreviations

HBSS:

Hank’s balanced salt solution

LTC4 :

Leukotriene C4

MRP:

Multidrug resistance related protein

OAT:

Organic anion transporter

PAPS:

3′-Phosphoadenosine 5′-phosphosulfate

SULT:

Sulfotransferases

UDPGA:

Uridine diphosphoglucuronic acid

UGT:

Uridine diphosphate-5′-glucuronosyltransferase or UDP-glucuronosyltransferase

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Author notes

  1. Jun Chen

    Present address: Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109-1065, USA

  2. Ming Hu

    Present address: Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX 77030, USA

Authors and Affiliations

  1. Department of Pharmaceutical Sciences, College of Pharmacy, Washington State University, Pullman, WA 99164-6510, USA

    Jun Chen, Huimin Lin & Ming Hu

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  1. Jun Chen
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  2. Huimin Lin
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  3. Ming Hu
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Correspondence to Ming Hu.

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Chen, J., Lin, H. & Hu, M. Absorption and metabolism of genistein and its five isoflavone analogs in the human intestinal Caco-2 model. Cancer Chemother Pharmacol 55, 159–169 (2005). https://doi.org/10.1007/s00280-004-0842-x

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  • DOI: https://doi.org/10.1007/s00280-004-0842-x

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