Steroid hormones specifically modify the activity of organic anion transporting polypeptides

doi:10.1016/j.ejps.2012.08.017

European Journal of Pharmaceutical Sciences

Volume 47, Issue 4, 20 November 2012, Pages 774-780

https://doi.org/10.1016/j.ejps.2012.08.017 Get rights and content

Abstract

Previously, the steroid hormone progesterone has been demonstrated to stimulate OATP2B1-mediated transport of estrone-3-sulphate (E₁S), dehydroepiandrosterone sulphate (DHEAS) and pregnenolone sulphate (PS), which may influence the uptake of precursor molecules for steroid hormone synthesis. However, it is unclear whether OATP2B1 drug substrates like atorvastatin or glibenclamide are also affected by this phenomenon. In addition, it has not been studied so far if this stimulatory effect is specific for OATP2B1. To address these questions, we examined the influence of progesterone on OATP2B1-mediated atorvastatin and glibenclamide uptake and studied the impact of steroid hormones on the transport activity of OATP1A2, OATP1B1 and OATP1B3.

Comparison of the substrate spectrum of the investigated OATPs revealed that DHEAS and atorvastatin are substrates of all transporters, while E₁S was only significantly transported by OATP1A2, OATP2B1 and OATP1B1. Glibenclamide uptake was limited to OATP1A2, OATP1B1 and OATP2’B1.

Subsequent interaction studies indicated that progesterone only increases OATP2B1-mediated E₁S and DHEAS transport, whereas uptake of BSP, atorvastatin and glibenclamide was either inhibited or not affected. Moreover, the steroid hormone effect was specific for OATP2B1; neither OATP1B1, OATP1B3 nor OATP1A2 function was stimulated in the presence of progesterone. Similar to progesterone, the glucocorticoide dexamethasone stimulated OATP2B1-mediated transport of E₁S and DHEAS (EC₅₀ for E₁S: 10.2 ± 5.6 μM and 17.9 ± 15.4 μM for DHEAS).

In conclusion, our data demonstrate that among the tested compounds the stimulatory effect of progesterone is specific for OATP2B1 and restricted to sulphated steroids like E₁S and DHEAS while the OATP-mediated drug transport is not enhanced.

Introduction

The organic anion transporting polypeptide (OATP/SLCO) family consists of 11 human members involved in the uptake of various endogenous and exogenous amphiphilic compounds. So far, pharmacological relevance has been demonstrated for OATP1A2, OATP1B1, OATP1B3 and OATP2B1 [for review see (Kalliokoski and Niemi, 2009, Muller and Fromm, 2011)]. Especially for OATP1B1 and OATP1B3, which are expressed in the sinusoidal membrane of the liver, numerous interaction and genetic studies have highlighted their role in hepatic drug uptake (Koenen et al., 2011, Meyer zu Schwabedissen and Kim, 2009). For example, it has been shown that genetic polymorphisms of OATP1B1 are associated with altered pharmacokinetics of statins (Romaine et al., 2010). In addition, there is evidence, that this transporter also affects efficacy and safety of drugs as shown for simvastatin (Couvert et al., 2008, Link et al., 2008). In contrast to the liver-specific OATP1B1, the expression of other OATP family members is not restricted to a single organ. For instance, besides liver, OATP1B3 expression has also been demonstrated in prostate and colon cancer (Hamada et al., 2008, Lee et al., 2008). OATP1A2 and OATP2B1 are expressed in a variety of tissues including the placenta, kidney, liver, intestine, heart and brain (Kalliokoski and Niemi, 2009). Numerous drugs have been shown to interact with these transporters. For OATP2B1, transport of the cholesterol-lowering agents/HMG-CoA reductase inhibitors atorvastatin and rosuvastatin, the antidiabetic drug glibenclamide and the leukotriene receptor antagonist montelukast as well as of several endogenous sulphate conjugates of steroid hormones has been demonstrated (Kalliokoski and Niemi, 2009, Mougey et al., 2009, Tamai et al., 2001).

As mentioned above, altered OATP function may affect disposition, efficacy and safety of drugs. Therefore, it is important to identify clinically relevant confounders of OATP function. Variability of transport function may arise from genetic polymorphisms, translational or posttranslational regulation as well as interactions of OATPs with modulators/inhibitors. In the latter case, we have previously reported that OATP2B1-mediated uptake of estrone-3-sulphate (E₁S), dehydroepiandrosterone sulphate (DHEAS) and pregnenolone sulphate (PS) is enhanced in the presence of the steroid hormone progesterone (Grube et al., 2006). This finding may be of special relevance for the formation of gestagens and estrogens in tissues like placenta and mammary gland, which are dependent on the uptake of precursor molecules for steroid hormone synthesis like PS or DHEAS. However, with regard to the proposed function of OATP2B1 as an uptake carrier for drugs into the enterocyte, the liver or organs like heart or placenta, modulation of transport – either by inhibition or stimulation – may substantially affect drug disposition. So far, it is still unknown whether the observed stimulatory effect of progesterone on OATP2B1-function is restricted to the transport of sulphated steroids or whether it also affects OATP2B1 drug substrates, like bromosulfophthalein (BSP), atorvastatin or glibenclamide. In addition, the influence of steroid hormones on OATP-function has only been investigated for OATP2B1 so far. We therefore tested whether progesterone and other steroid hormones also modifies the function of OATP1A2, 1B1 and 1B3, which share some substrates with OATP2B1.

Section snippets

Materials

[³H]atorvastatin (specific activity 10 Ci/mmol) was purchased from American Radiolabeled Chemicals (St. Louis, USA), [³H]E₁S (specific activity 50 Ci/mmol), [³H]BSP (specific activity 14 Ci/mmol) and [³H]DHEAS (specific activity 94.5 Ci/mmol) were from Hartmann Analytic (Braunschweig, Germany) and [³H]glibenclamide (specific activity 42 Ci/mmol) was from PerkinElmer Life and Analytical Sciences (Boston, USA). All other substances were obtained from Sigma–Aldrich (St. Louis, MO, USA).

Cell culture

HEK293 cells

Uptake of typical OATP2B1 substrates by OATP2B1, 1A2, 1B1 and 1B3

To compare the transport properties between OATP2B1 and the other OATPs, we studied the uptake of atorvastatin, BSP, DHEAS, E₁S and glibenclamide into OATP2B1-, 1A2-, 1B1- and 1B3-transfected HEK293 cells. Compared to vector transfected control cells, DHEAS was accumulated by OATP1A2 about 4.0-fold, by OATP2B1 1.5-fold (not significant), by OATP1B1 3.2-fold and 2.2-fold by OATP1B3. BSP and atorvastatin were taken up by almost all studied transporter proteins. However, compared to the other

Discussion

The organic anion transporting polypeptide (OATP) family mediates the cellular uptake of amphiphilic organic anions including bile acids, steroid hormone derivates as well as various drugs. Pharmacological relevance regarding drug interactions with liver specific OATPs has been demonstrated for instance for statins, repaglinide and the antihistamine fexofenadine (for review see (Kalliokoski and Niemi, 2009)). Knowledge about factors influencing OATP-function like genetic polymorphisms,

Conclusions

Taken together, we could demonstrate that the stimulatory effect of progesterone on OATP2B1 function is largely specific for the transport of sulphated steroids, while the transport of OATP2B1 drug substrates like atorvastatin or glibenclamide is inhibited or unaffected by progesterone. Moreover, the stimulatory effect of both progesterone and dexamethasone is specific for OATP2B1. The underlying mechanism of these findings is still unclear; however, our study indicates that there is a specific

Acknowledgments

We acknowledge the excellent technical assistance of Tina Sonnenberger (Department of Pharmacology, Ernst-Moritz-Arndt University, Greifswald, Germany). In addition we gratefully acknowledge Dr. Hartmut Glaeser and Dr. Kathrin Mandery, Institute of Experimental and Clinical Pharmacology and Toxicology, Friedrich-Alexander University, Erlangen, Germany, for providing the OATP1A2 cDNA.

This work was supported by a Grant of the Deutsche Forschungsgemeinschaft (Grant: GR 3375/1-1) and in part by the

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¹: These authors contributed equally to this work.

²: Present address: University of North Carolina, Eshelman School of Pharmacy, Chapel Hill, NC, USA².

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Steroid hormones specifically modify the activity of organic anion transporting polypeptides

Abstract

Introduction

Section snippets

Materials

Cell culture

Uptake of typical OATP2B1 substrates by OATP2B1, 1A2, 1B1 and 1B3

Discussion

Conclusions

Acknowledgments

J. Biol. Chem.

Eur. J. Pharmacol.

Gastroenterology

Biochem. Pharmacol.

J. Steroid Biochem. Mol. Biol.

Drug. Metab. Pharmacokinet

J. Biol. Chem.

Interaction of oral antidiabetic drugs with hepatic uptake transporters – focus on organic anion transporting polypeptides and organic cation transporter 1

Diabetes

Association of fetal hormone levels with stem cell potential: evidence for early life roots of human cancer

Cancer Res.

Patterns of sex steroid hormones in nipple aspirate fluid during the menstrual cycle and after menopause in relation to serum concentrations

Cancer Epidemiol. Biomarkers Prev.