Separation and detection of enantiomers of stilbestrol analogues by combined high-performance liquid chromatography—thermospray mass spectrometry

Parker, Carol E.; Levy, Louis; Smith, Richard; Yamaguchi, Kenji; Gaskell, S J; Korach, Kenneth S.

doi:10.1016/s0378-4347(00)82044-7

Cited by 11 publications

(5 citation statements)

References 11 publications

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“…This is in contrast to yeast-expressed glucocorticoid receptor where the dose-responses to various glucocorticoids were substantially altered compared with glucocorticoid receptor expressed in mammalian cells (Shena & Yamamoto 1988). In the present study, we used the yeast expression system to elucidate the structure-function relationship Our data demonstrate that the relative activities of DES-related compounds in yeast cells transfected with the mER are similar to the stimulation of uterine estrogen responses by these compounds (Korach et al 1985). The mER binds DES and IA-S with similar affinity yet DES has 7-5-fold greater transcriptional activity.…”

Section: Discussionmentioning

confidence: 73%

“…IA and DMIA were synthesized by ChemSyn Laboratories (Lenexa, KS, USA) as previously described (Korach et al 1979). Separation of the IA enantiomers was performed as described (Parker et al 1985). Oxalyticase was obtained from Enzogenetics (Corvallis, OR, USA).…”

Section: Methodsmentioning

confidence: 99%

“…The latter compound was of particular interest because of its high binding affinity to mouse uterine ER but weak uterotropic activity (Korach et al 1985). It was determined that IA has a single chiral carbon atom at C3 and exists as a mixture of enantiomers, IA-S and IA-R, in solution ( Fig.…”

mentioning

confidence: 99%

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Mutational analysis of the estrogen receptor ligand-binding domain: influence of ligand structure and stereochemistry on transactivation

Kohno

Bocchinfuso²,

Gandini³

et al. 1996

Journal of Molecular Endocrinology

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The mouse estrogen receptor (mER) exhibits ligand stereochemical specificity for indenestrol A (IA), a stilbestrol estrogen. IA has a chiral C3 methyl group, and the mER preferentially binds the S-enantiomer (IA-S), resulting in elevated biological activity when compared with the IA-R enantiomer. To elucidate the mechanisms for this stereochemical recognition, we have constructed a series of mERs with individual amino acid substitutions at Met521, His528, Met532, and Val537. The abilities of yeast-expressed wild-type and mutant mERs to transactivate an estrogen-responsive reporter gene construct were measured in the presence of diethylstilbestrol (DES) and IA enantiomers. The concentration of IA-S required to induce half-maximal transactivation by wild-type mER was 10-fold lower than IA-R, which is attributed to the 15-fold greater binding affinity for IA-S. Wild-type mER displayed similar dose-response curves for IA-R and demethyl IA, which lacks a C3 methyl group, demonstrating that the presence and correct orientation of the C3 methyl group on the IA compound is required for high-affinity ligand binding and transcriptional activity. Each mutant exhibited a reduced preference for IA-S enantiomer with respect to transactivation, suggesting that this region of the mER functions in ligand stereochemical recognition and activation. A mutation at Met532 diminished DES- and IA-S-induced transactivation by 7.5-fold and 40-fold respectively, with minimal change on their binding affinity. These data suggest that Met532 is required for transactivation induced by the potent agonist, IA-S, and the M532G mutation effectively uncouples IA-S ligand binding from transactivation. Use of these stereochemically different ligands in combination with mutagenesis of the mER demonstrates that ligand structure could influence transactivation by specifically altering the conformation of the mER AF-2 region.

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Section: Discussionmentioning

confidence: 73%

Section: Methodsmentioning

confidence: 99%

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Mutational analysis of the estrogen receptor ligand-binding domain: influence of ligand structure and stereochemistry on transactivation

Kohno

Bocchinfuso²,

Gandini³

et al. 1996

Journal of Molecular Endocrinology

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“…Because the target indene structure contains a stereocenter at C-1, all compounds were prepared as racemic mixtures. 38,[40][41][42] The core 3-ethyl-2-arylindene scaffold (4) was synthesized by Lewis-acid induced cyclodehydration of a ketone intermediate (3), following adaptations to a published synthetic route 43,44 (Scheme 1). Boron tribromide was used for the cyclodehydration of 3, because of its chelating effect, which sterically prevented formation of undesirable isomers obtained using polyphosphoric acid.…”

Section: Resultsmentioning

confidence: 99%

“…The 1-position of indenestrol A (Figure b) was already known to accommodate an alkyl extension of up to at least four carbons, without loss in binding affinity, , indicating a prime target site for derivatization with aryl-based extensions. Because the target indene structure contains a stereocenter at C-1, all compounds were prepared as racemic mixtures. ,− The core 3-ethyl-2-arylindene scaffold ( 4 ) was synthesized by Lewis-acid induced cyclodehydration of a ketone intermediate ( 3 ), following adaptations to a published synthetic route , (Scheme ). Boron tribromide was used for the cyclodehydration of 3 , because of its chelating effect, which sterically prevented formation of undesirable isomers obtained using polyphosphoric acid.…”

Section: Resultsmentioning

confidence: 99%

Differential Response of Estrogen Receptor Subtypes to 1,3-Diarylindene and 2,3-Diarylindene Ligands

et al. 2005

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Estrogen receptors (ERs) control transcription of genes important for normal human development and reproduction. The signaling networks are complex, and there is a need for a molecular level understanding of the roles of receptor subtypes ERalpha and ERbeta in normal physiology and as therapeutic targets. We synthesized two series of ER ligands, based on a common indene scaffold, in an attempt to develop compounds that can selectively modulate ER-mediated transcription. The 3-ethyl-1,2-diarylindenes, utilizing an amide linker for the 1-aryl extension, bind weakly to the ERs. The 2,3-diarylindenes bind with high affinity to the ER subtypes and demonstrate a range of different biological activities, both in transcriptional reporter gene assays and inhibition of estradiol-stimulated proliferation of MCF-7 cells. Several ligands differentiate between ERalpha and ERbeta subtypes at an estrogen response element (ERE), displaying various levels of partial to full agonist activity at ERalpha, while antagonizing estradiol action at ERbeta.

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Combined liquid chromatography mass spectrometry. Part III. Applications of thermospray

Arpino

1992

Mass Spectrometry Reviews

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In Part I1 (9), the basic instrument setup of the thermospray (TS) interface was described. This device was originally designed as a general direct coupling technique to a liquid chromatograph (LC/TS/MS), but it is often used in a direct flow injection mode (TS / MS). General operating principles, optimization strategies, and possible ionization mechanisms were also reviewed and commented on in Part 11. This third part in the series of reviews dedicated to LC / MS coupling techniques covers analytical applications using a thermospray interface. The covered literature being particularly abundant, inorganic applications when using a thermospray nebulizer as a sampling device for spectroscopic methods other than mass spectrometry (for example, atomic absorption or emission spectroscopy, inductively coupled plasma spectroscopy, etc.) and direct coupling of the TS interface to supercritical fluid chromatography have been purposely omitted. The same convention for the numbering system as in Part I and I1 is again adopted in this text. The term "thermospray" is unambiguous only when it covers a mechanical device that samples a liquid solution introduced into a mass spectrometer. Consequently, LC /TS/MS was purposely chosen, instead of other frequently used abbreviations and expressions such as thermospray LC/MS, TS-LC/MS, etc., because LC/TS/MS correctly describes the logical order of the chained instruments or techniques used.

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Separation and detection of enantiomers of stilbestrol analogues by combined high-performance liquid chromatography—thermospray mass spectrometry

Cited by 11 publications

References 11 publications

Mutational analysis of the estrogen receptor ligand-binding domain: influence of ligand structure and stereochemistry on transactivation

Mutational analysis of the estrogen receptor ligand-binding domain: influence of ligand structure and stereochemistry on transactivation

Differential Response of Estrogen Receptor Subtypes to 1,3-Diarylindene and 2,3-Diarylindene Ligands

Combined liquid chromatography mass spectrometry. Part III. Applications of thermospray

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