Dissecting Physiological Roles of Estrogen Receptor α and β with Potent Selective Ligands from Structure-Based Design

The distinct roles of the two estrogen receptor (ER) isotypes, ERα and ERβ, in mediating the physiological responses to estrogens are not completely understood. Although knockout animal experiments have been aiding to gain insight into estrogen signaling, additional information on the function of ERα and ERβ will be provided by the application of isotype-selective ER agonists. Based on the crystal structure of the ERα ligand binding domain and a homology model of the ERβ-ligand binding domain, we have designed steroidal ligands that exploit the differences in size and flexibility of the two ligand binding cavities. Compounds predicted to bind preferentially to either ERα or ERβ were synthesized and tested in vitro using radio-ligand competition and transactivation assays. This approach directly led to highly ER isotype-selective (∼200-fold) and potent ligands. To unravel physiological roles of the two receptors, in vivo experiments with rats were conducted using the ERα- and ERβ-selective agonists in comp...

[1]  B. Katzenellenbogen,et al.  Molecular basis for the subtype discrimination of the estrogen receptor-beta-selective ligand, diarylpropionitrile. , 2003, Molecular endocrinology.

[2]  E. Baulieu,et al.  Androgen and estrogen receptors in rat ventral prostate epithelium and stroma. , 1981, Endocrinology.

[3]  J. D. Neill,et al.  The Physiology of reproduction , 1988 .

[4]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[5]  B. Komm,et al.  Evaluation of an Estrogen Receptor-β Agonist in Animal Models of Human Disease , 2003 .

[6]  J. Gustafsson,et al.  Role of estrogen receptor beta in estrogen action. , 2001, Annual review of physiology.

[7]  J. Gustafsson,et al.  The novel estrogen receptor‐β subtype: potential role in the cell‐ and promoter‐specific actions of estrogens and anti‐estrogens , 1997, FEBS letters.

[8]  E. Slater,et al.  Characterization of the novel progestin gestodene by receptor binding studies and transactivation assays. , 1995, Contraception.

[9]  H. Lee,et al.  A PCR Analysis of ERα and ERβ mRNA Abundance in Rats and the Effect of Ovariectomy , 1999 .

[10]  J. Gorski,et al.  Relationship of estrogen receptors and protein synthesis to the mitogenic effect of estrogens. , 1985, Endocrinology.

[11]  K. Grandien,et al.  Printed in U.S.A. Copyright © 1997 by The Endocrine Society Comparison of the Ligand Binding Specificity and Transcript Tissue Distribution of Estrogen Receptors � and � , 2022 .

[12]  K. Griffiths,et al.  Inhibition of 5 alpha-reductase in genital skin fibroblasts and prostate tissue by dietary lignans and isoflavonoids. , 1995, The Journal of endocrinology.

[13]  J. Gustafsson,et al.  Cloning of a novel receptor expressed in rat prostate and ovary. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Alexander Hillisch,et al.  The role of protein 3D-structures in the drug discovery process. , 2003, EXS.

[15]  J. Corton,et al.  Interaction of Estrogenic Chemicals and Phytoestrogens with Estrogen Receptor β. , 1998, Endocrinology.

[16]  M. Bajpai,et al.  Involvement of tyrosine kinase and cAMP-dependent kinase cross-talk in the regulation of human sperm motility. , 2003, Reproduction.

[17]  J. Gustafsson,et al.  Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors alpha and beta. , 1997, Endocrinology.

[18]  Zbigniew Dauter,et al.  Molecular basis of agonism and antagonism in the oestrogen receptor , 1997, Nature.

[19]  K. Dechering,et al.  Estrogen receptors alpha and beta: two receptors of a kind? , 2000, Current medicinal chemistry.

[20]  L. Andersson,et al.  A role for estrogen receptor β in the regulation of growth of the ventral prostate , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[21]  T. Halgren.  Merck molecular force field. I. Basis, form, scope, parameterization, and performance of MMFF94 , 1996, J. Comput. Chem..

[22]  K. Fritzemeier,et al.  Estrogen action on hepatic synthesis of angiotensinogen and IGF-I: Direct and indirect estrogen effects , 1994, The Journal of Steroid Biochemistry and Molecular Biology.

[23]  P. Chambon,et al.  Effect of single and compound knockouts of estrogen receptors alpha (ERalpha) and beta (ERbeta) on mouse reproductive phenotypes. , 2000, Development.

[24]  P B Sigler,et al.  Crystallographic comparison of the estrogen and progesterone receptor's ligand binding domains. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[25]  P. Chambon,et al.  Effect of single and compound knockouts of estrogen receptors α (ERα) and β (ERβ) on mouse reproductive phenotypes , 2000 .

[26]  John A. Katzenellenbogen,et al.  Allosteric Control of Ligand Selectivity between Estrogen Receptors α and β - Implications for Other Nuclear Receptors , 2004 .

[27]  B. Katzenellenbogen,et al.  Estrogen receptor-beta potency-selective ligands: structure-activity relationship studies of diarylpropionitriles and their acetylene and polar analogues. , 2001, Journal of medicinal chemistry.

[28]  B. Katzenellenbogen,et al.  Pyrazole ligands: structure-affinity/activity relationships and estrogen receptor-alpha-selective agonists. , 2000, Journal of medicinal chemistry.

[29]  J. Gustafsson,et al.  The estrogen receptor family. , 1999, Current opinion in obstetrics & gynecology.

[30]  P. Sigler,et al.  Atomic structure of progesterone complexed with its receptor , 1998, Nature.

[31]  R. Hilgenfeld,et al.  Modern Methods of Drug Discovery , 2003, EXS.

[32]  K. Korach,et al.  Receptor null mice reveal contrasting roles for estrogen receptor α and β in reproductive tissues , 2000, The Journal of Steroid Biochemistry and Molecular Biology.

[33]  K. Dechering,et al.  Estrogen Receptors α and β : Two Receptors of a Kind ? , 2001 .

[34]  J. Gustafsson,et al.  Generation and reproductive phenotypes of mice lacking estrogen receptor beta. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[35]  M Carlquist,et al.  Structure of the ligand‐binding domain of oestrogen receptor beta in the presence of a partial agonist and a full antagonist , 1999, The EMBO journal.

[36]  Impact of isotype-selective estrogen receptor agonists on ovarian function. , 2004 .

[37]  C. Hegele-hartung,et al.  In Vitro and In Vivo Models to Characterise Estrogens and Antiestrogens , 1999 .

[38]  K. Dahlman-Wright,et al.  Estrogen Receptor (ER)-β Reduces ERα-Regulated Gene Transcription, Supporting a “Ying Yang” Relationship between ERα and ERβ in Mice , 2003 .

[39]  K. Korach,et al.  Estrogen receptor gene disruption: molecular characterization and experimental and clinical phenotypes. , 1996, Recent progress in hormone research.

[40]  J. Thomsen,et al.  Mechanisms of estrogen action. , 2001, Physiological reviews.

[41]  B. Katzenellenbogen,et al.  Activities of estrogen receptor alpha- and beta-selective ligands at diverse estrogen responsive gene sites mediating transactivation or transrepression , 2003, Molecular and Cellular Endocrinology.

[42]  B. Katzenellenbogen,et al.  Characterization of the Biological Roles of the Estrogen Receptors, ERα and ERβ, in Estrogen Target Tissues in Vivo through the Use of an ERα-Selective Ligand , 2002 .

[43]  J. Frasor,et al.  Response-Specific and Ligand Dose-Dependent Modulation of Estrogen Receptor (ER) α Activity by ERβ in the Uterus , 2003 .