Competitive Androgen Receptor Antagonism as a Factor Determining the Predictability of Cumulative Anti­androgenic Effects of Widely Used Pesticides

Frances Orton,¹ Erika Rosivatz,² Martin Scholze,¹ and Andreas Kortenkamp¹

¹Centre for Toxicology, School of Pharmacy, London, United Kingdom; ²Institute of Chemical Biology, Imperial College London, London, United Kingdom


Abstract

Background: Many pesticides in current use have recently been revealed as in vitro androgen receptor (AR) antagonists, but information about their combined effects is lacking.


Objective: We investigated the combined effects and the competitive AR antagonism of pesticide mixtures.


Methods: We used the MDA-kb2 assay to test a combination of eight AR antagonists that did not also possess AR agonist properties (“pure” antagonists; 8 mix: fludioxonil, fenhexamid, ortho-­phenylphenol, imazalil, tebuconazole, dimetho­morph, methiocarb, pirimiphos-methyl), a combina­tion of five AR antagonists that also showed agonist activity (5 mix: cyprodinil, pyrimethanil, vinclozolin, chlor­propham, linuron), and all pesticides combined (13 mix). We used concentration addition (CA) and independent action (IA) to formu­late additivity expectations, and Schild plot analyses to investigate competitive AR antagonism.


Results: A good agreement between the effects of the mixture of eight “pure” AR antagonists and the responses predicted by CA was observed. Schild plot analysis revealed that the 8 mix acted by competi­tive AR antagonism. However, the observed responses of the 5 mix and the 13 mix fell within the “prediction window” boundaries defined by the predicted regression curves of CA and IA. Schild plot analysis with these mixtures yielded anomalous responses incompatible with competitive receptor antagonism.


Conclusions: A mixture of widely used pesticides can, in a predictable manner, produce combined AR antagonist effects that exceed the responses elicited by the most potent component alone. Inasmuch as large populations are regularly exposed to mixtures of anti­androgenic pesticides, our results underline the need for considering combination effects for these substances in regulatory practice.


Key words: antiandrogen, AR-antagonism, concentration addition, endocrine disruption, fungicide, mixture, pesticide. 


Environ Health Perspect 120:1578–1584 (2012). http://dx.doi.org/10.1289/ehp.1205391 [Online 10 September 2012]


Address correspondence to F. Orton, Institute for the Environment, Brunel University, Kingston Lane, Uxbridge UB83PH UK. Telephone: 01895267208. Fax: 01895269761. E-mail: frances.orton@brunel.ac.uk


Supplemental Material is available online (http://dx.doi.org/10.1289/ehp.1205391).


This work was funded by the European Commission, FP7 programme (CONTAMED, grant 212502).


The authors declare they have no actual or potential competing financial interests.


Received 26 April 2012; Accepted 10 September 2012; Online 10 September 2012.