Structure–Activity Relationship of para-Carborane Selective Estrogen Receptor β AgonistsClick to copy article linkArticle link copied!
- David SedlákDavid SedlákCZ-OPENSCREEN, Institute of Molecular Genetics of the Czech Academy of Sciences, Vídeňská 1083, 142 20 Prague, Czech RepublicMore by David Sedlák
- Tyler A. WilsonTyler A. WilsonMedicinal Chemistry Shared Resource, Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210, United StatesMore by Tyler A. Wilson
- Werner TjarksWerner TjarksDivision of Medicinal Chemistry and Pharmacognosy College of Pharmacy, The Ohio State University, Columbus, Ohio 43210, United StatesMore by Werner Tjarks
- Hanna S. RadomskaHanna S. RadomskaDivision of Pharmaceutics and Pharmacology, College of Pharmacy, The Ohio State University, Columbus, Ohio 43210, United StatesMore by Hanna S. Radomska
- Hongyan WangHongyan WangDivision of Pharmaceutics and Pharmacology, College of Pharmacy, The Ohio State University, Columbus, Ohio 43210, United StatesMore by Hongyan Wang
- Jayaprakash Narayana KollaJayaprakash Narayana KollaCZ-OPENSCREEN, Institute of Molecular Genetics of the Czech Academy of Sciences, Vídeňská 1083, 142 20 Prague, Czech RepublicMore by Jayaprakash Narayana Kolla
- Zbigniew J. LeśnikowskiZbigniew J. LeśnikowskiLaboratory of Medicinal Chemistry, Institute of Medical Biology PAS, 106 Lodowa Street, 93-232 Lodz, PolandMore by Zbigniew J. Leśnikowski
- Alena ŠpičákováAlena ŠpičákováDepartment of Pharmacology, Faculty of Medicine, Palacky University, Hněvotínská 3, 77515 Olomouc, Czech RepublicMore by Alena Špičáková
- Tehane AliTehane AliDivision of Medicinal Chemistry and Pharmacognosy College of Pharmacy, The Ohio State University, Columbus, Ohio 43210, United StatesMore by Tehane Ali
- Keisuke IshitaKeisuke IshitaDivision of Medicinal Chemistry and Pharmacognosy College of Pharmacy, The Ohio State University, Columbus, Ohio 43210, United StatesMore by Keisuke Ishita
- Liva Harinantenaina RakotondraibeLiva Harinantenaina RakotondraibeDivision of Medicinal Chemistry and Pharmacognosy College of Pharmacy, The Ohio State University, Columbus, Ohio 43210, United States
- Sandip VibhuteSandip VibhuteMedicinal Chemistry Shared Resource, Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210, United StatesMore by Sandip Vibhute
- Dasheng WangDasheng WangMedicinal Chemistry Shared Resource, Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210, United StatesMore by Dasheng Wang
- Pavel AnzenbacherPavel AnzenbacherDepartment of Pharmacology, Faculty of Medicine, Palacky University, Hněvotínská 3, 77515 Olomouc, Czech RepublicMore by Pavel Anzenbacher
- Chad BennettChad BennettMedicinal Chemistry Shared Resource, Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210, United StatesDrug Development Institute, Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210, United StatesMore by Chad Bennett
- Petr Bartunek*Petr Bartunek*Email: [email protected]CZ-OPENSCREEN, Institute of Molecular Genetics of the Czech Academy of Sciences, Vídeňská 1083, 142 20 Prague, Czech RepublicMore by Petr Bartunek
- Christopher C. Coss*Christopher C. Coss*Email: [email protected]. Tel: 614-688-1309.Division of Pharmaceutics and Pharmacology, College of Pharmacy, The Ohio State University, Columbus, Ohio 43210, United StatesDrug Development Institute, Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210, United StatesMore by Christopher C. Coss
Abstract
Selective agonism of the estrogen receptor (ER) subtypes, ERα and ERβ, has historically been difficult to achieve due to the high degree of ligand-binding domain structural similarity. Multiple efforts have focused on the use of classical organic scaffolds to model 17β-estradiol geometry in the design of ERβ selective agonists, with several proceeding to various stages of clinical development. Carborane scaffolds offer many unique advantages including the potential for novel ligand/receptor interactions but remain relatively unexplored. We synthesized a series of para-carborane estrogen receptor agonists revealing an ERβ selective structure–activity relationship. We report ERβ agonists with low nanomolar potency, greater than 200-fold selectivity for ERβ over ERα, limited off-target activity against other nuclear receptors, and only sparse CYP450 inhibition at very high micromolar concentrations. The pharmacological properties of our para-carborane ERβ selective agonists measure favorably against clinically developed ERβ agonists and support further evaluation of carborane-based selective estrogen receptor modulators.
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Introduction
Figure 1
Figure 1. Estrogen receptor ligands.
Results and Discussion
Chemistry
Scheme 1
a(a) n-BuLi, dimethoxyethane (1,2-DME), −78 or 0 °C; (b) BBr3, CH2CI2, 0 °C to room temperature (RT).
Scheme 2
a(a) PCC, CH2Cl2; (b) BH3·THF, (S)-2-Me-CBS, RT; (c) BH3·THF, (R)-2-Me-CBS, RT; (d) BBr3, CH2Cl2, 0 °C to RT.
Scheme 3
a(a) n-Hexylmagnesium bromide, Et2O, 0 °C; (b) PCC, CH2CI2; (c) BH3·THF, (R)-2-Me-CBS, 0 °C; (d) 1-dodecanethiol, N-methylpyrrolidinone (NMP), NaOH, 100 °C.
Scheme 4
a(a) Cs2CO3, CH3I, acetone, reflux; (b) (methoxymethyl)triphenylphosphonium chloride, LiHMDS, THF; 2 N HCI; (c) hexylmagnesium bromide, Et2O, 0 °C; (d) PCC, CH2CI2; (e) BH3·THF, (R)-2-Me-CBS, 0°C; (f) 1-dodecanethiol, NMP, NaOH, 100 °C.
Scheme 5
a(a) Br2, dichloromethane (DCM), HgO; (b) AICI3, benzene, (c) LiAIH4, Et2O; (d) PCC, NaHCO3, NaOAc, CH2CI2; (e) hexylmagnesium bromide, Et2O; (f) AgOAc, Br2, CHCI3; (g) PCC, CH2CI2; (h) BH3·THF, (R)-2-Me-CBS 0 °C; (i) benzaldehyde oxime, Cs2CO3, RockPhos Pd G3.
Scheme 6
a(a) n-BuLi, 1,2-DME, −78 or 0 °C, 1-heptanal; (b) BBr3, CH2CI2, 0 °C to RT.
Structure–Activity Relationship (SAR) Study

Each compound was tested for ERα and ERβ agonism in luciferase transactivation assay and compared to 17β-estradiol. The EC50 values are shown in nanomolar. Agonism is reported as full (≥67%) colored in green, partial (34−66%) colored in yellow, weak partial (10−33%) colored in red, and no agonism (<10%) not colored. To determine the ranking of agonism, percent agonism was calculated using Emax for each compound and comparing to the activity of 100 nM 17β-estradiol set to 100%.
ERβ selectivity was calculated as the ratio of EC50 for ERα/EC50 for ERβ.
Compound cytotoxicity from parallel experiment to reporter assays determined by measurement of intracellular adenosine 5′-triphosphate (ATP) levels. IC50 values are expressed in micromolar.
Compound affinities for ERα or ERβ proteins were evaluated in polarized fluorescence-based competitive binding assay PolarScreen ER α/β. Compounds were allowed to compete with Fluormone EL Red ligand for the binding to the receptor in a dose response experiment, and Kd values were calculated from IC50 values using the Cheng−Prusoff equation.
n.a., not active.
n.c., not calculated.
Figure 2
Figure 2. Ligand selectivity for ERβ increases with elongation of the side hydrocarbon chain. BE120, 3g, and 8 were tested in ERα (dashed line) and ERβ (solid line) transactivation assays in the concentration range between 10 μM and 1 pM, in triplicates and activities were expressed relative to 100 nM 17β-estradiol set to 100%. Increasing length of the carboranes’ side hydrocarbon chain is accompanied with decreasing potency in both ERα and ERβ reporter assays and by increasing selectivity for ERβ (blue: nonselective, green: 10–100× selective, red: selectivity >100×).

Each compound was tested for ERα and ERβ agonism in luciferase transactivation assay and compared to 17β-estradiol. The EC50 values are shown in nanomolar. Agonism is reported as full (≥67%) colored in green, partial (34−66%) colored in yellow, weak partial (10−33%) colored in red, and no agonism (<10%) not colored. To determine the ranking of agonism, percent agonism was calculated using Emax for each compound and comparing to the activity of 100 nM 17β-estradiol set to 100%.
ERβ selectivity was calculated as the ratio of EC50 for ERα/EC50 for ERβ.
Compound cytotoxicity from parallel experiment to reporter assays determined by measurement of intracellular adenosine 5′-triphosphate (ATP) levels. IC50 values are expressed in micromolar.
Compound affinities for ERα or ERβ proteins were evaluated in polarized fluorescence-based competitive binding assay PolarScreen ER α/β. Compounds were allowed to compete with Fluormone EL Red ligand for the binding to the receptor in a dose response experiment, and Kd values were calculated from IC50 values using the Cheng−Prusoff equation.

Each compound was tested for ERα and ERβ agonism in luciferase transactivation assay and compared to 17β-estradiol. The EC50 values are shown in nanomolar. Agonism is reported as full (≥67%) colored in green, partial (34−66%) colored in yellow, weak partial (10−33%) colored in red, and no agonism (<10%) not colored. To determine the ranking of agonism, percent agonism was calculated using Emax for each compound and comparing to the activity of 100 nM 17β-estradiol set to 100%.
ERβ selectivity was calculated as the ratio of EC50 for ERα/EC50 for ERβ.
Compound cytotoxicity from parallel experiment to reporter assays determined by measurement of intracellular adenosine 5′-triphosphate (ATP) levels. IC50 values are expressed in micromolar.
Compound affinities for ERα or ERβ proteins were evaluated in polarized fluorescence-based competitive binding assay PolarScreen ER α/β. Compounds were allowed to compete with Fluormone EL Red ligand for the binding to the receptor in a dose response experiment, and Kd values were calculated from IC50 values using the Cheng−Prusoff equation.
n.c., not calculated.
n.d., not determined.

Each compound was tested for ERα and ERβ agonism in luciferase transactivation assay and compared to 17β-estradiol. The EC50 values are shown in nanomolar. Agonism is reported as full (≥67%) colored in green, partial (34–66%) colored in yellow, weak partial (10–33%) colored in red, and no agonism (<10%) not colored. To determine the ranking of agonism, percent agonism was calculated using Emax for each compound and comparing to the activity of 100 nM 17β-estradiol set to 100%.
ERβ selectivity was calculated as the ratio of EC50 for ERα/EC50 for ERβ.
Compound cytotoxicity from parallel experiment to reporter assays determined by measurement of intracellular adenosine 5′-triphosphate (ATP) levels. IC50 values are expressed in micromolar.
Compound affinities for ERα or ERβ proteins were evaluated in polarized fluorescence-based competitive binding assay PolarScreen ER α/β. Compounds were allowed to compete with Fluormone EL Red ligand for the binding to the receptor in a dose response experiment, and Kd values were calculated from IC50 values using the Cheng–Prusoff equation.
n.a., not active.
n.c., not calculated.
Interactions with Other Nuclear Receptors (NRs)
Figure 3
Figure 3. Activities of the carboranes’ compound library on steroid receptors. (A) Compounds were profiled at 1 μM in the selective luciferase reporter assays for the agonistic and (B) antagonistic activities for steroid receptors. The activity is expressed relative to maximal activity induced by reference compound or as a fold induction compared to untreated cells for control assay (MMTV reporter alone, dark red dots). Cell viability in U2OS cells was used as a control experiment for the antagonistic activities and is expressed in a scale of 0–100% relative to dimethyl sulfoxide (DMSO)-treated cells (green dots).
Inhibition of Cytochrome 450 Isoforms

Inhibitory activities of compounds were tested in dose response experiment with specific substrates for individual CYP forms. The enzyme activity was detected by HPLC Prominence system (Shimadzu; Kyoto, Japan) with a UV/fluorescence detection.
n.a., not active.
Conclusions
Experimental Section
Chemicals, Materials, and Methods
Synthesis
1-(4-Methoxyphenyl)-12-ethyl-1,12-dicarba-closo-dodecaborane (2b)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]ethan-1-ol (2c)
1-(4-Methoxyphenyl)-12-propyl-1,12-dicarba-closo-dodecaborane (2d)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]propan-1-ol (2e)
1-(4-Methoxyphenyl)-12-butyl-1,12-dicarba-closo-dodecaborane (2f)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]butan-1-ol (2g)
1-(4-Methoxyphenyl)-12-pentyl-1,12-dicarba-closo-dodecaborane (2h)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]pentan-1-ol (2i)
1-(4-Methoxyphenyl)-12-heptyl-1,12-dicarba-closo-dodecaborane (2j)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]heptan-1-ol (2k)
1-(4-Methoxyphenyl)-12-decyl-1,12-dicarba-closo-dodecaborane (2l)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]decyl-1-ol (2m)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-4-methylpentan-1-ol (2n)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-5-methylhexan-1-ol (2o)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-6-methylheptan-1-ol (2q)
(RS)-(2,3-Dihydro-1H-inden-5-yl)-[1-(4-methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]methanol (2r)
1-(4-Methoxyphenyl)-12-(3-phenylpropyl)-1,12-dicarba-closo-dodecaborane (2s)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-3-phenylpropan-1-ol (2t)
1-(4-Methoxyphenyl)-12-(4-phenylbutyl)-1,12-dicarba-closo-dodecaborane (2u)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-4-phenylbutan-1-ol (2v)
1-(4-Methoxyphenyl)-12-(5-phenylpentyl)-1,12-dicarba-closo-dodecaborane (2w)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-5-phenylpentan-1-ol (2x)
1-(4-Hydroxyphenyl)-12-ethyl-1,12-dicarba-closo-dodecaborane (3b)
(RS)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]ethan-1-ol (3c)
1-(4-Hydroxyphenyl)-12-propyl-1,12-dicarba-closo-dodecaborane (3d)
(RS)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]propan-1-ol (3e)
1-(4-Hydroxyphenyl)-12-butyl-1,12-dicarba-closo-dodecaborane (3f)
(RS)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]butan-1-ol (3g)
1-(4-Hydroxyphenyl)-12-pentyl-1,12-dicarba-closo-dodecaborane (3h)
(RS)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]pentan-1-ol (3i)
1-(4-Hydroxyphenyl)-12-heptyl-1,12-dicarba-closo-dodecaborane (3j)
(RS)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]heptan-1-ol (3k)
1-(4-Hydroxyphenyl)-12-decyl-1,12-dicarba-closo-dodecaborane (3l)
(RS)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]decan-1-ol (3m)
(RS)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-4-methylpentan-1-ol (3n)
(RS)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-5-methylhexan-1-ol (3o)
(RS)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-6-methylheptan-1-ol (3q)
(RS)-(2,3-Dihydro-1H-inden-5-yl)-[1-(4-hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]methanol (3r)
1-(4-Hydroxyphenyl)-12-(3-phenylpropyl)-1,12-dicarba-closo-dodecaborane (3s)
(RS)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-3-phenylpropan-1-ol (3t)
1-(4-Hydroxyphenyl)-12-(4-phenylbutyl)-1,12-dicarba-closo-dodecaborane (3u)
(RS)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-4-phenylbutan-1-ol (3v)
1-(4-Hydroxyphenyl)-12-(-phenylpentyl)-1,12-dicarba-closo-dodecaborane (3w)
(RS)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]-5-phenylpentan-1-ol (3x)
1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]heptan-1-one (4)
(S)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]heptan-1-ol (5)
(R)-1-[1-(4-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]heptan-1-ol (6)
(S)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]heptan-1-ol (7)
(R)-1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]heptan-1-ol (8)
1-[1-(4-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]heptan-1-one (9)
1-(4′-Methoxy-[1,1′-biphenyl]-4-yl)heptan-1-one (11)
(S)-1-(4′-Methoxy-[1,1′-biphenyl]-4-yl)heptan-1-ol (12)
(S)-4′-(1-Hydroxyheptyl)-[1,1′-biphenyl]-4-ol (13)
4-(4-Methoxyphenyl)cyclohexan-1-one (15)
1-(4-(4-Methoxyphenyl)cyclohexyl)heptan-1-one (16)
(S)-4-(4-(1-Hydroxyheptyl)cyclohexyl)phenol (17)
Methyl 4-Bromobicyclo[2.2.2]octane-1-carboxylate (19)
Methyl 4-Phenylbicyclo[2.2.2]octane-1-carboxylate (20)
4-Phenylbicyclo[2.2.2]octane-1-carbaldehyde (21)
1-(4-Phenylbicyclo[2.2.2]octan-1-yl)heptan-1-ol (22)
1-(4-(4-Bromophenyl)bicyclo[2.2.2]octan-1-yl)heptan-1-ol (23)
1-(4-(4-Bromophenyl)bicyclo[2.2.2]octan-1-yl)heptan-1-one (24)
(S)-1-(4-(4-Bromophenyl)bicyclo[2.2.2]octan-1-yl)heptan-1-ol (25)
(S)-4-(4-(1-Hydroxyheptyl)bicyclo[2.2.2]octan-1-yl)phenol (26)
(RS)-1-[(1-Phenyl)-1,12-dicarba-closo-dodecaborane-12-yl]heptan-1-ol (28)
(RS)-1-[1-(3-Methoxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]heptan-1-ol (30)
(RS)-1-[1-(3-Hydroxyphenyl)-1,12-dicarba-closo-dodecaborane-12-yl]heptan-1-ol (31)
Plasmids and Transfection
Reporter Luciferase Assays for Steroid Receptors AR, PR, GR, and MR
Ga4/UAS Reporter Luciferase Assays for Nuclear Receptors
Luciferase Reporter Assays
Cell Viability Assay
PolarScreen ERα and ERβ Competitor Binding Assay
Cytochrome P450 Enzymatic Assays
Supporting Information
The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.jmedchem.1c00555.
Activity profiles of library compounds in cell-based reporter assays for nuclear receptors; methods and experimental conditions for cytochrome P450 enzyme assays; and NMR and MS spectra and HPLC chromatograms (PDF)
Molecular formula strings (CSV)
Terms & Conditions
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Acknowledgments
This work was supported by the Ministry of Education, Youth and Sports grant: RVO: 68378050-KAV-NPUI, LM2015063, LM2018130, NSC 2014/13/B/NZ1/03989 at IMB PAS, institutional support RVO: 68378050 at IMG and RVO: 61989592 at UPOL. This work was further supported by the College of Pharmacy at The Ohio State University, its instrumentation core and director Dr. Craig McElroy. This study made use of the Campus Chemical Instrument Center NMR facility at The Ohio State University. Compounds 13, 17, and 26 were synthesized by the Medicinal Chemistry Shared Resource, and the corresponding mass spectral data were obtained by the Proteomics Shared Resource, both of which are part of The Ohio State University Comprehensive Cancer Center and supported by NCI/NIH Grant P30CA016058. This work was also supported by the Drug Development Institute within The Ohio State University Comprehensive Cancer Center and Pelotonia. The authors thank the University of Illinois Urbana–Champaign Mass Spectrometry Laboratory and the University of Michigan College of Literature, Science and the Arts Mass Spectrometry Technical Services for instrumentation support. They also thank Olga Martínková for support in cell culture work and transfection experiments, Dr. Martin Popr for support in compound management, Tomas Langammer for automating the experiments on HTS robotics, and Dr. Samuel Kulp for his help in formatting the manuscript.
AR | androgen receptor |
BNCT | boron-neutron capture therapy |
CBS | Corey–Bakshi–Shibata |
CDCl3 | deuterated chloroform |
DBD | DNA-binding domain |
DMEM | Dulbecco’s modified Eagle’s medium |
DPN | diarylpropionitrile |
E2 | 17β-estradiol |
ERRγ | estrogen receptor-related receptor γ |
ERα | estrogen receptor α |
ERβ | estrogen receptor β |
EtOAc | ethyl acetate |
EtOH | ethyl alcohol |
FBS | fetal bovine serum |
FXR | farnesoid X receptor |
GPER1 | G protein-coupled transmembrane receptor |
GR | glucocorticoid receptor |
i-PrOH | isopropyl alcohol |
KO | knock-out |
LTR | long terminal repeat |
LXR | liver X receptor |
MMTV | mouse mammary tumor virus |
MR | mineralocorticoid receptor |
NaOAc | sodium acetate |
NIMBA | N-isopropyl-N-methyl-tert-butylamine |
NR | nuclear receptor |
PPAR | peroxisome proliferator-activated receptor |
PPT | propylpyrazole triol |
PR | progesterone receptor |
RAR | retinoic acid receptor |
RXR | retinoid X receptor |
SR | steroid receptor |
UAS | upstream activating sequences |
VDR | vitamin D receptor |
Additional Notes
a Preparation of 5-Methylhexan-1-al: PCC (3.71 g, 17.21 mmol) was suspended in dry methylene chloride (40 mL). A solution of 5-methyl-1-hexanol (1 g, 8.61 mmol) in methylene chloride (15 mL) was then added in one portion to the stirred suspension to give a dark reaction mixture, which was stirred at room temperature overnight. Dry ether (60 mL) was added and then molecular sieve followed by stirring for 1 h. The supernatant liquid was decanted, and the insoluble residue was washed with dry ether (3 × 20 mL). The combined organic phases were passed through a short florisil column, and the solvent was removed in vacuo providing 370 mg (38%) of product, which was used without further purification and analysis in the next step.
b Preparation of 6-Methylheptan-1-al: PCC (2.89 g, 13.44 mmol) was suspended in dry methylene chloride (40 mL). A solution of 6-methyl-1-heptanol (1 g, 7.68 mmol) in methylene chloride (15 mL) was then added in one portion to the stirred suspension to give a dark reaction mixture, which was stirred at room temperature overnight. Dry ether (60 mL) was added and then molecular sieve followed by stirring for 1 h. The supernatant liquid was decanted, and the insoluble residue was washed with dry ether (3 × 20 mL). The combined organic phases were passed through a short florisil column, and the solvent was removed in vacuo providing 750 mg (76%) of product, which was used without further purification and analysis in the next step.
References
This article references 70 other publications.
- 1Dodds, E. C.; Lawson, W. Synthetic Strogenic Agents without the Phenanthrene Nucleus. Nature 1936, 137, 996, DOI: 10.1038/137996a0Google Scholar1https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaA28XkvVagsQ%253D%253D&md5=9991eb1f0557091ee8f4d26b2c9d99d3Synthetic estrogenic agents without the phenanthrene nucleusDodds, E. C.; Lawson, WilfridNature (London, United Kingdom) (1936), 137 (), 996CODEN: NATUAS; ISSN:0028-0836.The phenanthrene condensed-ring structure is not necessary for estrogenic activity. A table of 14 synthetic substances is given, all except one showing 100% positive estrus responses.
- 2Walter, P.; Green, S.; Greene, G.; Krust, A.; Bornert, J. M.; Jeltsch, J. M.; Staub, A.; Jensen, E.; Scrace, G.; Waterfield, M. Cloning of the Human Estrogen Receptor CDNA. Proc. Natl. Acad. Sci. U.S.A. 1985, 82, 7889– 7893, DOI: 10.1073/pnas.82.23.7889Google Scholar2https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaL28XotlShsA%253D%253D&md5=bfb5de85940113236c09e1f34a785b75Cloning of the human estrogen receptor cDNAWalter, Philippe; Green, Stephen; Greene, Geoffrey; Krust, Andree; Bornert, Jean Marc; Jeltsch, Jean Marc; Staub, Adrien; Jensen, Elwood; Scrace, Geoffrey; et al.Proceedings of the National Academy of Sciences of the United States of America (1985), 82 (23), 7889-93CODEN: PNASA6; ISSN:0027-8424.Poly(A)+ RNA isolated from the human breast cancer cell line MCF-7 was fractionated by sucrose gradient centrifugation and fractions enriched in estrogen receptor (ER) mRNA were used to prep. randomly primed cDNA libraries in the λg10 and λg11 vectors. Clones corresponding to ER sequence were isolated from both libraries after screening with either ER monoclonal antibodies (λg11) or synthetic oligonucleotide probes designed from 2 peptide sequences of purified ER (λg10). Five cDNA clones were isolated by antibody screening, and 5 were isolated after screening with synthetic oligonucleotides. The 2 largest ER cDNA clones, λOR3 (1.3 kilobase pairs) and λOR8 (2.1 kilobase pairs), isolated by using antibodies and oligonucleotides, resp., were able to enrich selectively for ER mRNA by hybrid-selection. Further, λOR8 contains the DNA sequence expected from the 2 ER peptides and cross-hybridizes with each of the other ER cDNA clones. These results demonstrate that the clones isolated correspond to the ER mRNA sequence. Use of λOR8 as a hybridization probe revealed a single poly(A)+ RNA band of ≈6.2 kilobase pairs in the ER-contg. human breast cancer cell lines MCF-7 and T47D. In contrast, no hybridization was seen in the human ER-neg. cell line HeLa. The same probe hybridizes to a chicken gene that is expressed in oviduct tissue as a 7.5-kilobase-pair poly(A)+ RNA.
- 3Korach, K. S.; Couse, J. F.; Curtis, S. W.; Washburn, T. F.; Lindzey, J.; Kimbro, K. S.; Eddy, E. M.; Migliaccio, S.; Snedeker, S. M.; Lubahn, D. B.; Schomberg, D. W.; Smith, E. P. Estrogen Receptor Gene Disruption: Molecular Characterization and Experimental and Clinical Phenotypes. Recent Prog. Horm. Res. 1996, 51, 159– 186Google Scholar3https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2sXktFKisr0%253D&md5=b9f0b64fca403a8ac04eff2884ad258aEstrogen receptor gene disruption: molecular characterization and experimental and clinical phenotypesKorach, Kenneth S.; Couse, John F.; Curtis, Sylvia W.; Washburn, Todd F.; Lindzey, Jonathan; Kimbro, K. Sean; Eddy, Edward M.; Migliaccio, Silvia; Snedeker, Suzanne M.; Lubahn, Dennis B.; Schomberg, David W.; Smith, Eric P.Recent Progress in Hormone Research (1996), 51 (), 159-188CODEN: RPHRA6; ISSN:0079-9963. (Endocrine Society)A review with 50 refs. The estrogen receptor (ER) is thought to play a crucial role in the regulation of many life processes, including development, reprodn. and normal physiol. Because there have been no known mutations of the estrogen receptor in normal tissue of humans and animals, its presence and tissue distribution is thought to be essential for survival. Using the techniques of homologous recombination, the ER gene has been disrupted to produce a line of transgenic mice possessing the altered ER gene (ERKO). The mouse ER gene was disrupted by inserting a 1.8-kb PGK-Neomycin sequence into exon 2, ∼280 bp downstream of the transcription start codon. The correct targeting of the disruption was demonstrated by Southern blot anal. and PCR. Western blot anal. of uterine prepns. from ERKO females showed no detectable ER protein. Heterozygotes had one half the level of ER protein compared to wild-type animals. Estrogen insensitivity was confirmed using estrogen agonists, estradiol, hydroxy tamoxifen, diethylstilbestrol treatment for 3 days which resulted in a 3-4-fold increase in uterine wet wt. and vaginal cornification in wild-type females, while ERKO mice were totally unresponsive. These data were further supported by the failure of estrogen or EGF treatment to induce DNA synthesis in uterine tissue of similarly treated mice. Lactoferrin, an estrogen-responsive gene in the uterus, was also assayed by Northern blot. Wild-type mice treated with a single estradiol injection showed a 350-fold induction in lactoferrin mRNA, while ERKO females showed no detectable response. Both male and female animals survive to adulthood with normal gross external phenotypes. As expected, females are infertile and demonstrate hypoplastic uteri and hyperemic ovaries with no apparent corpora lutea. Males are also infertile, with atrophy of the testes and seminiferous tubule dysmorphogenesis. Although the reproductive capabilities have been altered with a dramatic effect on the gonads, prenatal development of the reproductive tracts of both sexes appear to be independent of an ER-mediated response. Anal. of the mammary glands of the ERKO females at 4 mo of age showed a primitive ductal rudiment rather than the fully developed ductal tree seen in wild-type siblings. Also absent were the terminal end buds seen during normal ductal morphogenesis. Both sexes show a decrease in skeletal bone d., supporting a direct role for ER action in bone. A single patient is described who is homozygous for a point mutation in the human ER gene at codon 157. The mutation produces a truncation of the ER protein and results in estrogen insensitivity syndrome. Most significant of the clin. findings are effects on skeletal bone d. and retarded bone age. Findings from the patient and mice suggest that the absence of functional ER is not lethal. Mutation in the ER gene is present in the human population. Further characterization of the mice and identification of addnl. patients will be required to more fully understand the consequences of ER gene mutations.
discussion 186–188.
- 4Kuiper, G. G.; Enmark, E.; Pelto-Huikko, M.; Nilsson, S.; Gustafsson, J. A. Cloning of a Novel Receptor Expressed in Rat Prostate and Ovary. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 5925– 5930, DOI: 10.1073/pnas.93.12.5925Google Scholar4https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK28Xjs1Ojs7c%253D&md5=aa1c112de706d680f59afec3e77830b3Cloning of a novel estrogen receptor expressed in rat prostate and ovaryKuiper, George G. J. M.; Enmark, Eva; Pelto-Huikko, Markku; Nilsson, Stefan; Gustafsson, Jan-AakeProceedings of the National Academy of Sciences of the United States of America (1996), 93 (12), 5925-5930CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)The authors have cloned a novel member of the nuclear receptor superfamily. The cDNA of clone 29 was isolated from a rat prostate cDNA library and it encodes a protein of 485 amino acid residues with a calcd. mol. wt. of 54.2 kDa. Clone 29 protein is unique in that it is highly homologous to the rat estrogen receptor (ER) protein, particularly in the DNA-binding domain (95%) and in the C-terminal ligand-binding domain (55%). Expression of clone 29 in rat tissues was investigated by in situ hybridization and prominent expression was found in prostate and ovary. In the prostate clone 29 is expressed in the epithelial cells of the secretory alveoli, whereas in the ovary the granulosa cells in primary, secondary, and mature follicles showed expression of clone 29. Satn. ligand-binding anal. of in vitro synthesized clone 29 protein revealed a single binding component for 17β-estradiol (E2) and high affinity (Kd = 0.6 nM). In ligand-competition expts. the binding affinity decreased in the order E2 > diethylstilbestrol > estriol > estrone > 5α-androstane-3β,17β-diol » testosterone = progesterone = corticosterone = 5α-androstane-3α,17β-diol. In cotransfection expts. of Chinese hamster ovary cells with a clone 29 expression vector and an estrogen-regulated reporter gene, maximal stimulation (about 3-fold) of reporter gene activity was found during incubation with 10 nM of E2. Neither progesterone, testosterone, dexamethasone, thyroid hormone, all-trans-retinoic acid, nor 5α-androstane-3α,17β-diol could stimulate reporter gene activity, whereas estrone and 5α-androstane-3β,17β-diol did. The authors conclude that clone 29 cDNA encodes a novel rat ER, which they suggest be named rat ERβ to distinguish it from the previously cloned ER (ERα) from rat uterus.
- 5Nilsson, S.; Mäkelä, S.; Treuter, E.; Tujague, M.; Thomsen, J.; Andersson, G.; Enmark, E.; Pettersson, K.; Warner, M.; Gustafsson, J.-Å. Mechanisms of Estrogen Action. Physiol. Rev. 2001, 81, 1535– 1565, DOI: 10.1152/physrev.2001.81.4.1535Google Scholar5https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD38XislCqtw%253D%253D&md5=e3c660fbdf99d1b456486caabef516c8Mechanisms of estrogen actionNilsson, Stefan; Makela, Sari; Treuter, Eckardt; Tujague, Michel; Thomsen, Jane; Andersson, Goran; Enmark, Eva; Pettersson, Katarina; Warner, Margaret; Gustafsson, Jan-AkePhysiological Reviews (2001), 81 (4), 1535-1565CODEN: PHREA7; ISSN:0031-9333. (American Physiological Society)A review. Our appreciation of the physiol. functions of estrogens and the mechanisms through which estrogens bring about these functions has changed during the past decade. Just as transgenic mice were produced in which estrogen receptors had been inactivated and we thought that we were about to understand the role of estrogen receptors in physiol. and pathol., it was found that there was not 1 but 2 distinct and functional estrogen receptors, now called ERα and ERβ. Transgenic mice in which each of the receptors or both the receptors are inactive have revealed a much broader role for estrogens in the body than was previously thought. This decade also saw the description of a male patient who had no functional ERα and whose continued bone growth clearly revealed an important function of estrogen in men. The importance of estrogen in both males and females was also demonstrated in the lab. in transgenic mice in which the aromatase gene was inactivated. Finally, crystal structures of the estrogen receptors with agonists and antagonists have revealed much about how ligand binding influences receptor conformation and how this conformation influences interaction of the receptor with coactivators or corepressors and hence dets. cellular response to ligands.
- 6Heldring, N.; Pike, A.; Andersson, S.; Matthews, J.; Cheng, G.; Hartman, J.; Tujague, M.; Ström, A.; Treuter, E.; Warner, M.; Gustafsson, J.-Å. Estrogen Receptors: How Do They Signal and What Are Their Targets. Physiol. Rev. 2007, 87, 905– 931, DOI: 10.1152/physrev.00026.2006Google Scholar6https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXptFGls7k%253D&md5=207d16fa37d6b14483b9fe1002ce2646Estrogen receptors: how do they signal and what are their targetsHeldring, Nina; Pike, Ashley; Andersson, Sandra; Matthews, Jason; Cheng, Guojun; Hartman, Johan; Tujague, Michel; Stroem, Anders; Treuter, Eckardt; Warner, Margaret; Gustafsson, Jan-AakePhysiological Reviews (2007), 87 (3), 905-931CODEN: PHREA7; ISSN:0031-9333. (American Physiological Society)A review. During the past decade there has been a substantial advance in our understanding of estrogen signaling both from a clin. as well as a preclin. perspective. Estrogen signaling is a balance between two opposing forces in the form of two distinct receptors (ERα and ERβ) and their splice variants. The prospect that these two pathways can be selectively stimulated or inhibited with subtype-selective drugs constitutes new and promising therapeutic opportunities in clin. areas as diverse as hormone replacement, autoimmune diseases, prostate and breast cancer, and depression. Mol. biol., biochem., and structural studies have generated information which is invaluable for the development of more selective and effective ER ligands. We have also become aware that ERs do not function by themselves but require a no. of coregulatory proteins whose cell-specific expression explains some of the distinct cellular actions of estrogen. Estrogen is an important morphogen, and many of its proliferative effects on the epithelial compartment of glands are mediated by growth factors secreted from the stromal compartment. Thus understanding the cross-talk between growth factor and estrogen signaling is essential for understanding both normal and malignant growth. In this review we focus on several of the interesting recent discoveries concerning estrogen receptors, on estrogen as a morphogen, and on the mol. mechanisms of anti-estrogen signaling.
- 7Carmeci, C.; Thompson, D. A.; Ring, H. Z.; Francke, U.; Weigel, R. J. Identification of a Gene (GPR30) with Homology to the G-Protein-Coupled Receptor Superfamily Associated with Estrogen Receptor Expression in Breast Cancer. Genomics 1997, 45, 607– 617, DOI: 10.1006/geno.1997.4972Google Scholar7https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2sXnsVejsbo%253D&md5=831bb2d172786fe61c914e8e851d0735Identification of a gene (GPR30) with homology to the G-protein-coupled receptor superfamily associated with estrogen receptor expression in breast cancerCarmeci, Charles; Thompson, Devon A.; Ring, Huijun Z.; Francke, Uta; Weigel, Ronald J.Genomics (1997), 45 (3), 607-617CODEN: GNMCEP; ISSN:0888-7543. (Academic Press)Using the technique of differential cDNA library screening, a cDNA clone was isolated from an estrogen receptor (ER)-pos. breast carcinoma cell line (MCF7) cDNA library based upon the overexpression of this gene compared to an ER-neg. cell line (MDA-MB-231). Sequence anal. of this clone detd. that it shared significant homol. to G-protein-coupled receptors. This receptor, GPCR-Br, was abundantly expressed in the ER-pos. breast carcinoma cell lines MCF7, T-47D, and MDA-MB-361. Expression was absent or minimal in the ER-neg. breast carcinoma cell lines BT-20, MDA-MB-231, and HBL-100. GPCR-Br was ubiquitously expressed in human tissues examd. but was most abundant in placenta. GPCR-Br expression was examd. in 11 primary breast carcinomas. GPCR-Br was detected in all 4 ER-pos. tumors and only 1 of 7 ER-neg. tumors. Based upon PCR anal. in hybrid cell lines, the gene for GPCR-Br (HGMW-approved symbol GPR30) was mapped to chromosome 7p22. The pattern of expression of GPCR-Br indicates that this receptor may be involved in physiol. responses specific to hormonally responsive tissues.
- 8Revankar, C. M.; Cimino, D. F.; Sklar, L. A.; Arterburn, J. B.; Prossnitz, E. R. A Transmembrane Intracellular Estrogen Receptor Mediates Rapid Cell Signaling. Science 2005, 307, 1625– 1630, DOI: 10.1126/science.1106943Google Scholar8https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXitV2is7c%253D&md5=938d5a0422146eac0ace28da892d82efA Transmembrane Intracellular Estrogen Receptor Mediates Rapid Cell SignalingRevankar, Chetana M.; Cimino, Daniel F.; Sklar, Larry A.; Arterburn, Jeffrey B.; Prossnitz, Eric R.Science (Washington, DC, United States) (2005), 307 (5715), 1625-1630CODEN: SCIEAS; ISSN:0036-8075. (American Association for the Advancement of Science)The steroid hormone estrogen regulates many functionally unrelated processes in numerous tissues. Although it is traditionally thought to control transcriptional activation through the classical nuclear estrogen receptors, it also initiates many rapid nongenomic signaling events. We found that of all G protein-coupled receptors characterized to date, GPR30 is uniquely localized to the endoplasmic reticulum, where it specifically binds estrogen and fluorescent estrogen derivs. Activating GPR30 by estrogen resulted in intracellular calcium mobilization and synthesis of phosphatidylinositol 3,4,5-trisphosphate in the nucleus. Thus, GPR30 represents an intracellular transmembrane estrogen receptor that may contribute to normal estrogen physiol. as well as pathophysiol.
- 9Kuiper, G. G. J. M.; Carlsson, B.; Grandien, K.; Enmark, E.; Häggblad, J.; Nilsson, S.; Gustafsson, J.-Å. Comparison of the Ligand Binding Specificity and Transcript Tissue Distribution of Estrogen Receptors α and β. Endocrinology 1997, 138, 863– 870, DOI: 10.1210/endo.138.3.4979Google Scholar9https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2sXht1OksrY%253D&md5=bb4d88e7ba9c878a79c711b3b0a5f92fComparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors α and βKuiper, George G. J. M.; Carlsson, Bo; Grandien, Kaj; Enmark, Eva; Haeggblad, Johan; Nilsson, Stefan; Gustafsson, Jan-AakeEndocrinology (1997), 138 (3), 863-870CODEN: ENDOAO; ISSN:0013-7227. (Endocrine Society)The rat estrogen receptor (ER) exists as two subtypes, ERα and ERβ, which differ in the C-terminal ligand-binding domain and in the N-terminal transactivation domain. In this study we investigated the mRNA expression of both ER subtypes in rat tissues by RT-PCR and compared the ligand binding specificity of the ER subtypes. Satn. ligand binding anal. of in vitro synthesized human ERα and rat ERβ protein revealed a single binding component for 16α-iodo-17β-estradiol with high affinity [dissocn. const. (Kd) = 0.1 nM for ERα protein and 0.4 nM for ERβ protein]. Most estrogenic substrates or estrogenic antagonists compete with 16α-[125I]iodo-17β-estradiol for binding to both ER subtypes in a very similar preference and degree; i.e., diethylstilbestrol > hexestrol > dienestrol > 4-OH-tamoxifen > 17β-estradiol > coumestrol, ICI-164384 > estrone, 17α-estradiol > nafoxidine, moxestrol > clomifene > estriol, 4-OH-estradiol > tamoxifen, 2-OH-estradiol, 5-androstene-3β-17β-diol, genistein for the ERα protein and dienestrol > 4-OH-tamoxifen > diethylstilbestrol > hexestrol > coumestrol, ICI-164384 > 17β-estradiol > estrone, genistein > estriol > nafoxidine, 5-androstene-3β,17β-diol > 17α-estradiol, clomifene, 2-OH-estradiol > 4-OH-estradiol, tamoxifen, moxestrol for the ERβ protein. The rat tissue distribution and/or the relative level of ERα and ERβ expression seems to be quite different, i.e. moderate to high expression in uterus, testis, pituitary, ovary, kidney, epididymis, and adrenal for ERα and prostate, ovary, lung, bladder, brain, uterus, and testis for ERβ. The described differences between the ER subtypes in relative ligand binding affinity and tissue distribution could contribute to the selective action of ER agonists and antagonists in different tissues.
- 10Hamilton, K. J.; Hewitt, S. C.; Arao, Y.; Korach, K. S. Estrogen Hormone Biology. In Current Topics in Developmental Biology; Forrest, D.; Tsai, S., Eds.; Nuclear Receptors in Development and Disease; Academic Press: 2017; Chapter 4; Vol. 125, pp 109– 146.Google ScholarThere is no corresponding record for this reference.
- 11Zimmerman, M. A.; Budish, R. A.; Kashyap, S.; Lindsey, S. H. GPER–Novel Membrane Oestrogen Receptor. Clin. Sci. 2016, 130, 1005– 1016, DOI: 10.1042/CS20160114Google Scholar11https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XotVSrsLs%253D&md5=7404f65583d895b9ccc19505eb9a858bGPER-novel membrane estrogen receptorZimmerman, Margaret A.; Budish, Rebecca A.; Kashyap, Shreya; Lindsey, Sarah H.Clinical Science (2016), 130 (12), 1005-1016CODEN: CSCIAE; ISSN:1470-8736. (Portland Press Ltd.)The recent discovery of the G protein-coupled estrogen receptor (GPER) presents new challenges and opportunities for understanding the physiol., pathophysiol. and pharmacol. of many diseases. This review will focus on the expression and function of GPER in hypertension, kidney disease, atherosclerosis, vascular remodelling, heart failure, reprodn., metabolic disorders, cancer, environmental health and menopause. Furthermore, this review will highlight the potential of GPER as a therapeutic target.
- 12Mäkelä, S.; Savolainen, H.; Aavik, E.; Myllärniemi, M.; Strauss, L.; Taskinen, E.; Gustafsson, J.-Å.; Häyry, P. Differentiation between Vasculoprotective and Uterotrophic Effects of Ligands with Different Binding Affinities to Estrogen Receptors α and β. Proc. Natl. Acad. Sci. U.S.A. 1999, 96, 7077– 7082, DOI: 10.1073/pnas.96.12.7077Google Scholar12https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXks1WktbY%253D&md5=eac9076dd6e164de1216e54059d9782cDifferentiation between vasculoprotective and uterotrophic effects of ligands with different binding affinities to estrogen receptors α and βMakela, Sari; Savolainen, Hanna; Aavik, Einari; Myllarniemi, Marjukka; Strauss, Leena; Taskinen, Eero; Gustafsson, Jan-Ake; Hayry, PekkaProceedings of the National Academy of Sciences of the United States of America (1999), 96 (12), 7077-7082CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)Estrogen-based drug therapy in cardiovascular diseases has been difficult because it has not been possible to sep. the wanted vasculoprotective effect from the unwanted effects of the hormone to the reproductive system. Here we demonstrate that, after endothelial denudation of rat carotid artery, the mRNA of the classical estrogen receptor (ER) is constitutively expressed at a low level, whereas the expression of the novel ER mRNA increases >40-fold. Under in situ hybridization and immunohistochem., ERβ mRNA and protein colocalize with the smooth muscle cells in the media and neointima. Treatment of ovariectomized female rats with the isoflavone phytoestrogen genistein, which shows 20-fold higher binding affinity to ERβ than to ERα, or with 17β-estradiol, which does not differentiate between the two receptors, provides similar dose-dependent vasculoprotective effect in rat carotid injury model. In addn. in concn. <10 μM, both ligands are equally inhibitory to the replication and migration of smooth muscle cells in vitro. However, only treatment with 17β-estradiol, but not with genistein, is accompanied with a dose-dependent uterotrophic effect. The results suggest that preferential targeting to ERβ will provide vasculoprotective estrogen analogs devoid of effects to the reproductive system.
- 13Thomas, P.; Pang, Y.; Filardo, E. J.; Dong, J. Identity of an Estrogen Membrane Receptor Coupled to a G Protein in Human Breast Cancer Cells. Endocrinology 2005, 146, 624– 632, DOI: 10.1210/en.2004-1064Google Scholar13https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXotlWitw%253D%253D&md5=6870880ddbd84da144dc18d929707bcdIdentity of an estrogen membrane receptor coupled to a G protein in human breast cancer cellsThomas, P.; Pang, Y.; Filardo, E. J.; Dong, J.Endocrinology (2005), 146 (2), 624-632CODEN: ENDOAO; ISSN:0013-7227. (Endocrine Society)Although nonclassical estrogen actions initiated at the cell surface have been described in many tissues, the identities of the membrane estrogen receptors (mERs) mediating these actions remain unclear. Here we show that GPR30, an orphan receptor unrelated to nuclear estrogen receptors, has all the binding and signaling characteristics of a mER. A high-affinity (dissocn. const. 2.7 nM), limited capacity, displaceable, single binding site specific for estrogens was detected in plasma membranes of SKBR3 breast cancer cells that express GPR30 but lack nuclear estrogen receptors. Progesterone-induced increases and small interfering RNA-induced decreases in GPR30 expression in SKBR3 cells were accompanied by parallel changes in specific estradiol-17β (E2) binding. Plasma membranes of human embryonic kidney 293 cells transfected with GPR30, but not those of untransfected cells, and human placental tissues that express GPR30 also displayed high-affinity, specific estrogen binding typical of mERs. E2 treatment of transfected cell membranes caused activation of a stimulatory G protein that is directly coupled to the receptor, indicating GPR30 is a G protein-coupled receptor (GPCR), and also increased adenylyl cyclase activity. The finding that the antiestrogens tamoxifen and ICI 182,780, and an environmental estrogen, ortho, para-dichlorodiphenyl-dichloroethylene (o,p'-DDE), have high binding affinities to the receptor and mimic the actions of E2 has important implications for both the development and treatment of estrogen-dependent breast cancer. GPR30 is structurally unrelated to the recently discovered family of GPCR-like membrane progestin receptors. The identification of a second distinct class of GPCR-like steroid membrane receptors suggests a widespread role for GPCRs in nonclassical steroid hormone actions.
- 14Lubahn, D. B.; Moyer, J. S.; Golding, T. S.; Couse, J. F.; Korach, K. S.; Smithies, O. Alteration of Reproductive Function but Not Prenatal Sexual Development after Insertional Disruption of the Mouse Estrogen Receptor Gene. Proc. Natl. Acad. Sci. U.S.A. 1993, 90, 11162– 11166, DOI: 10.1073/pnas.90.23.11162Google Scholar14https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2cXks1ajsw%253D%253D&md5=bde5d2b52c3e2fda7c834175b3e61ca0Alteration of reproductive function but not prenatal sexual development after insertional disruption of the mouse estrogen receptor geneLubahn, Dennis B.; Moyer, Jeffrey S.; Golding, Thomas S.; Couse, John F.; Korach, Kenneth S.; Smithies, OliverProceedings of the National Academy of Sciences of the United States of America (1993), 90 (23), 11162-6CODEN: PNASA6; ISSN:0027-8424.Estrogen receptor and its ligand, estradiol, have long been thought to be essential for survival, fertility, and female sexual differentiation and development. Consistent with this proposed crucial role, no human estrogen receptor gene mutations are known, unlike the androgen receptor, where many loss of function mutations have been found. The authors have generated mutant mice lacking responsiveness to estradiol by disrupting the estrogen receptor gene by gene targeting. Both male and female animals survive to adulthood with normal gross external phenotypes. Females are infertile; males have a decreased fertility. Females have hypoplastic uteri and hyperemic ovaries with no detectable corpora lutea. In adult wild-type and heterozygous females, 3-day estradiol treatment at 40 μg/kg stimulates a 3- to 4-fold increase in uterine wet wt. and alters vaginal cornification, but the uteri and vagina do not respond in the animals with the estrogen receptor gene disruption. Prenatal male and female reproductive tract development can therefore occur in the absence of estradiol receptor-mediated responsiveness.
- 15Krege, J. H.; Hodgin, J. B.; Couse, J. F.; Enmark, E.; Warner, M.; Mahler, J. F.; Sar, M.; Korach, K. S.; Gustafsson, J.-Å.; Smithies, O. Generation and Reproductive Phenotypes of Mice Lacking Estrogen Receptor β. Proc. Natl. Acad. Sci. U.S.A. 1998, 95, 15677– 15682, DOI: 10.1073/pnas.95.26.15677Google Scholar15https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXhvFeiuw%253D%253D&md5=50cc5538ab791d52f280959e296d0e8dGeneration and reproductive phenotypes of mice lacking estrogen receptor βKrege, John H.; Hodgin, Jeffrey B.; Couse, John F.; Enmark, Eva; Warner, Margaret; Mahler, Joel F.; Sar, Madhabananda; Korach, Kenneth S.; Gustafsson, Jan-Ake; Smithies, OliverProceedings of the National Academy of Sciences of the United States of America (1998), 95 (26), 15677-15682CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)Estrogens influence the differentiation and maintenance of reproductive tissues and affect lipid metab. and bone remodeling. Two estrogen receptors (ERs) have been identified to date, ERα and ERβ. The authors previously generated and studied knockout mice lacking estrogen receptor α and reported severe reproductive and behavioral phenotypes including complete infertility of both male and female mice and absence of breast tissue development. Here, the authors describe the generation of mice lacking estrogen receptor β (ERβ -/-) by insertion of a neomycin-resistance gene into exon 3 of the coding gene by using homologous recombination in embryonic stem cells. Mice lacking this receptor develop normally and are indistinguishable grossly and histol. as young adults from their littermates. RNA anal. and immunocytochem. show that tissues from ERβ -/- mice lack normal ERβ RNA and protein. Breeding expts. with young, sexually mature females show that they are fertile and exhibit normal sexual behavior, but have fewer and smaller litters than wild-type mice. Superovulation expts. indicate that this redn. in fertility is the result of reduced ovarian efficiency. The mutant females have normal breast development and lactate normally. Young, sexually mature male mice show no overt abnormalities and reproduce normally. Older mutant males display signs of prostate and bladder hyperplasia. Thus, ERβ is essential for normal ovulation efficiency but is not essential for female or male sexual differentiation, fertility, or lactation. Future expts. are required to det. the role of ERβ in bone and cardiovascular homeostasis.
- 16Prossnitz, E. R.; Hathaway, H. J. What Have We Learned about GPER Function in Physiology and Disease from Knockout Mice?. J. Steroid Biochem. Mol. Biol. 2015, 153, 114– 126, DOI: 10.1016/j.jsbmb.2015.06.014Google Scholar16https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2MXht1CkurnN&md5=76ffa0a4124d62506b9add2f0459dd32What have we learned about GPER function in physiology and disease from knockout mice?Prossnitz, Eric R.; Hathaway, Helen J.Journal of Steroid Biochemistry and Molecular Biology (2015), 153 (), 114-126CODEN: JSBBEZ; ISSN:0960-0760. (Elsevier Ltd.)Estrogens, predominantly 17β-estradiol, exert diverse effects throughout the body in both normal and pathophysiol., during development and in reproductive, metabolic, endocrine, cardiovascular, nervous, musculoskeletal and immune systems. Estrogen and its receptors also play important roles in carcinogenesis and therapy, particularly for breast cancer. In addn. to the classical nuclear estrogen receptors (ERα and ERβ) that traditionally mediate predominantly genomic signaling, the G protein-coupled estrogen receptor GPER has become recognized as a crit. mediator of rapid signaling in response to estrogen. Mouse models, and in particular knockout (KO) mice, represent an important approach to understand the functions of receptors in normal physiol. and disease. Whereas ERα KO mice display multiple significant defects in reprodn. and mammary gland development, ERβ KO phenotypes are more limited, and GPER KO exhibit no reproductive deficits. However, the study of GPER KO mice over the last six years has revealed that GPER deficiency results in multiple physiol. alterations including obesity, cardiovascular dysfunction, insulin resistance and glucose intolerance. In addn., the lack of estrogen-mediated effects in numerous tissues of GPER KO mice, studied in vivo or ex vivo, including those of the cardiovascular, endocrine, nervous and immune systems, reveals GPER as a genuine mediator of estrogen action. Importantly, GPER KO mice have also demonstrated roles for GPER in breast carcinogenesis and metastasis. In combination with the supporting effects of GPER-selective ligands and GPER knockdown approaches, GPER KO mice demonstrate the therapeutic potential of targeting GPER activity in diseases as diverse as obesity, diabetes, multiple sclerosis, hypertension, atherosclerosis, myocardial infarction, stroke and cancer.
- 17Harris, H. A. Estrogen Receptor-β: Recent Lessons from in Vivo Studies. Mol. Endocrinol. 2007, 21, 1– 13, DOI: 10.1210/me.2005-0459Google Scholar17https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXmsFOnsw%253D%253D&md5=f55e31161a74e96fa82518f7aac9e62fEstrogen receptor-β: recent lessons from in vivo studiesHarris, Heather A.Molecular Endocrinology (2007), 21 (1), 1-13CODEN: MOENEN; ISSN:0888-8809. (Endocrine Society)A review. The unexpected discovery of a second form of the estrogen receptor (ER), designated ERβ, surprised and energized the field of estrogen research. In the 9 yr since its identification, the remarkable efforts from academic and industrial scientists of many disciplines have made significant progress in elucidating its biol. A powerful battery of tools, including knockout mice as well as a panel of receptor-selective agonists, has allowed an investigation into the role of ERβ. To date, in vivo efficacy studies are limited to rodents. Current data indicate that ERβ plays a minor role in mediating estrogen action in the uterus, on the hypothalamus/pituitary, the skeleton, and other classic estrogen target tissues. However, a clear role for ERβ has been established in the ovary, cardiovascular system, and brain as well as in several animal models of inflammation including arthritis, endometriosis, inflammatory bowel disease, and sepsis. The next phase of research will focus on elucidating, at a mol. level, how ERβ exerts these diverse effects and exploring the clin. utility of ERβ-selective agonists.
- 18Warner, M.; Wu, W.; Montanholi, L.; Nalvarte, I.; Antonson, P.; Gustafsson, J.-A. Ventral Prostate and Mammary Gland Phenotype in Mice with Complete Deletion of the ERβ Gene. Proc. Natl. Acad. Sci. U.S.A. 2020, 117, 4902– 4909, DOI: 10.1073/pnas.1920478117Google Scholar18https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXksVSrur0%253D&md5=5b9cf899ca7c8779041f68a6164349d1Ventral prostate and mammary gland phenotype in mice with complete deletion of the ERβ geneWarner, Margaret; Wu, Wan-fu; Montanholi, Leticia; Nalvarte, Ivan; Antonson, Per; Gustafsson, Jan-AkeProceedings of the National Academy of Sciences of the United States of America (2020), 117 (9), 4902-4909CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)Disagreements about the phenotype of estrogen receptor β; (ERβ) knockout mouse, created by removing the DNA-binding domain of the ERβ; gene or interruption of the gene with a neocassette (Oliver Smithies ERβ knockout mice [ERβOS-/-]), prompted us to create an ERβ knockout mouse by deleting the ERβ gene with the use of CRISPR/Cas9 technol. We confirmed that the ERβ gene was eliminated from the mouse genome and that no ERβ mRNA or protein was detectable in tissues of this mouse. Overall the phenotype of the ventral prostate (VP) and mammary gland (MG) in ERβcrispr-/- mice was similar to, but more severe than, that in the ERβOS-/-mice. In the VP of 6-mo-old ERβcrispr-/- mice there was epithelial hyperplasia, fibroplasia, inflammation, stromal overgrowth, and intraductal cancer-like lesions. This was accompanied by an increase in Ki67 and P63 and loss in DACH1 and PURα, two androgen receptor (AR) repressors. In the MG there was overexpression of estrogen receptor α and progesterone receptor, loss of collagen, increase in proliferation and expression of metalloproteases, and invasive epithelium. Surprisingly, by 18 mo of age, the no. of hyperplastic foci was reduced, the ducts of the VP and MG became atrophic, and, in the VP, there was massive immune infiltration and massive desquamation of the luminal epithelial cells. These changes were coincident with reduced levels of androgens in males and estrogens in females. We conclude that ERβ is a tumor suppressor gene in the VP and MG where its loss increases the activity AR and ERα resp.
- 19Meyers, M. J.; Sun, J.; Carlson, K. E.; Marriner, G. A.; Katzenellenbogen, B. S.; Katzenellenbogen, J. A. Estrogen Receptor-β Potency-Selective Ligands: Structure–Activity Relationship Studies of Diarylpropionitriles and Their Acetylene and Polar Analogues. J. Med. Chem. 2001, 44, 4230– 4251, DOI: 10.1021/jm010254aGoogle Scholar19https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3MXnsFagsr8%253D&md5=8fbf46c36aaf2ffcc9a8c44b15827bfeEstrogen Receptor-β Potency-Selective Ligands: Structure-Activity Relationship Studies of Diarylpropionitriles and Their Acetylene and Polar AnaloguesMeyers, Marvin J.; Sun, Jun; Carlson, Kathryn E.; Marriner, Gwendolyn A.; Katzenellenbogen, Benita S.; Katzenellenbogen, John A.Journal of Medicinal Chemistry (2001), 44 (24), 4230-4251CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)Through an effort to develop novel ligands that have subtype selectivity for the estrogen receptors alpha (ERα) and beta (ERβ), we have found that 2,3-bis(4-hydroxyphenyl)propionitrile (DPN) acts as an agonist on both ER subtypes, but has a 70-fold higher relative binding affinity and 170-fold higher relative potency in transcription assays with ERβ than with ERα. To investigate the ERβ affinity- and potency-selective character of this DPN further, we prepd. a series of DPN analogs in which both the ligand core and the arom. rings were modified by the repositioning of phenolic hydroxy groups and by the addn. of alkyl substituents and nitrile groups. We also prepd. other series of DPN analogs in which the nitrile functionality was replaced with acetylene groups or polar functions, to mimic the linear geometry or polarity of the nitrile, resp. To varying degrees, all of the analogs show preferential binding affinity for ERβ (i.e., they are ERβ affinity-selective), and many, but not all of them, are also more potent in activating transcription through ERβ than through ERα (i.e., they are ERβ potency-selective). meso-2,3-Bis(4-hydroxyphenyl)succinonitrile and dl-2,3-bis(4-hydroxyphenyl)succinonitrile are among the highest ERβ affinity-selective ligands, and they have an ERβ potency selectivity that is equiv. to that of DPN. The acetylene analogs have higher binding affinities but somewhat lower selectivities than their nitrile counterparts. The polar analogs have lower affinities, and only the fluorinated polar analogs have substantial affinity selectivities. This study suggests that, in this series of ligands, the nitrile functionality is crit. to ERβ selectivity because it provides the optimal combination of linear geometry and polarity. Furthermore, the addn. of a second nitrile group β to the nitrile in DPN or the addn. of a Me substituent at an ortho position on the β-arom. ring increases the affinity and selectivity of these compds. for ERβ. These ERβ-selective compds. may prove to be valuable tools in understanding the differences in structure and biol. function of ERα and ERβ.
- 20Stauffer, S. R.; Coletta, C. J.; Tedesco, R.; Nishiguchi, G.; Carlson, K.; Sun, J.; Katzenellenbogen, B. S.; Katzenellenbogen, J. A. Pyrazole Ligands: Structure–Affinity/Activity Relationships and Estrogen Receptor-α-Selective Agonists. J. Med. Chem. 2000, 43, 4934– 4947, DOI: 10.1021/jm000170mGoogle Scholar20https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3cXot1yjsbk%253D&md5=3bce249ad2d0eaf91aa4ad17ebffdd05Pyrazole Ligands: Structure-Affinity/Activity Relationships and Estrogen Receptor-α-Selective AgonistsStauffer, Shaun R.; Coletta, Christopher J.; Tedesco, Rosanna; Nishiguchi, Gisele; Carlson, Kathryn; Sun, Jun; Katzenellenbogen, Benita S.; Katzenellenbogen, John A.Journal of Medicinal Chemistry (2000), 43 (26), 4934-4947CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)We have found that certain tetrasubstituted pyrazoles are high-affinity ligands for the estrogen receptor (ER) and that one pyrazole is considerably more potent as an agonist on the ERα than on the ERβ subtype. To investigate what substituent pattern provides optimal ER binding affinity and the greatest enhancement of potency as an ERα-selective agonist, we prepd. a no. of tetrasubstituted pyrazole analogs with defined variations at certain substituent positions. Anal. of their binding affinity pattern shows that a C(4)-Pr substituent is optimal and that a p-hydroxyl group on the N(1)-Ph group also enhances affinity and selectivity for ERα. The best compd. in this series, a propylpyrazole triol (I), binds to ERα with high affinity (ca. 50% that of estradiol), and it has a 410-fold binding affinity preference for ERα. It also activates gene transcription only through ERα. Thus, this compd. represents the first ERα-specific agonist. We investigated the mol. basis for the exceptional ERα binding affinity and potency selectivity of pyrazole I by a further study of structure-affinity relationships in this series and by mol. modeling. These investigations suggest that the pyrazole triols prefer to bind to ERα with their C(3)-phenol in the estradiol A-ring binding pocket and that binding selectivity results from differences in the interaction of the pyrazole core and C(4)-Pr group with portions of the receptor where ERα has a smaller residue than ERβ. These ER subtype-specific interactions and the ER subtype-selective ligands that can be derived from them should prove useful in defining those biol. activities in estrogen target cells that can be selectively activated through ERα.
- 21Warner, M.; Huang, B.; Gustafsson, J.-A. Estrogen Receptor β as a Pharmaceutical Target. Trends Pharmacol. Sci. 2017, 38, 92– 99, DOI: 10.1016/j.tips.2016.10.006Google Scholar21https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XhslKnu7jM&md5=976f8a25342bba055e9ba332c84f7ff2Estrogen Receptor β as a Pharmaceutical TargetWarner, Margaret; Huang, Bo; Gustafsson, Jan-AkeTrends in Pharmacological Sciences (2017), 38 (1), 92-99CODEN: TPHSDY; ISSN:0165-6147. (Elsevier Ltd.)A major issue in clin. endocrinol. today is how to use hormones to achieve the health benefits that they clearly can provide but avoid the neg. side effects, i.e., how to develop more precise medicines. This problem of how to use hormones is pervasive in clin. endocrinol. It is true for estrogen, progesterone, androgen, vitamin D, and thyroid hormone, and the problem is amplified in the case of new ligands for the more recently discovered nuclear receptors. Selective targeting of hormone receptor subtypes is one attractive way to harness the beneficial effects of hormones while reducing unwanted side effects. Here, we focus on estrogen receptor (ER)β, which has promise as a selective target in hormone replacement therapy, and in breast and prostate cancers.
- 22Enmark, E.; Gustafsson, J.-Å. Oestrogen Receptors – an Overview. J. Intern. Med. 1999, 246, 133– 138, DOI: 10.1046/j.1365-2796.1999.00545.xGoogle Scholar22https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXmtlCmtb8%253D&md5=731a0a091639714ccb2a4b8760eec5a6Oestrogen receptors - an overviewEnmark, E.; Gustafsson, J.-A.Journal of Internal Medicine (1999), 246 (2), 133-138CODEN: JINMEO; ISSN:0954-6820. (Blackwell Science Ltd.)A review with 29 refs. The estrogen receptor (ER) is a ligand-activated transcription factor that mediates the effects of the steroid hormone 17β-estradiol in both males and females. Since the isolation and cloning of ER, the prevailing opinion has been that only one such receptor exists. The finding of a second subtype of ER (ERβ) has caused considerable excitement and has forced endocrinologists to re-evaluate many aspects of the actions of estrogens. In this article, the authors will try to summarize the current knowledge about the two estrogen receptor subtypes, with the emphasis on estrogen receptor β (ERβ), and to comment on the observations in mice lacking either receptor or the hormone itself.
- 23Harris, H. A.; Bapat, A. R.; Gonder, D. S.; Frail, D. E. The Ligand Binding Profiles of Estrogen Receptors α and β Are Species Dependent. Steroids 2002, 67, 379– 384, DOI: 10.1016/S0039-128X(01)00194-5Google Scholar23https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD38XhslWgs7g%253D&md5=7d8abb8e1b0083d859b19556dcfa4355The ligand binding profiles of estrogen receptors α and β are species dependentHarris, Heather A.; Bapat, Ashok R.; Gonder, Daniel S.; Frail, Donald E.Steroids (2002), 67 (5), 379-384CODEN: STEDAM; ISSN:0039-128X. (Elsevier Science Inc.)Estrogens and selective estrogen receptor modulators are used for the treatment and prevention of conditions resulting from menopause. Since estrogens exert their activity by binding to nuclear receptors, there is intense interest in developing new ligands for the two known estrogen receptor subtypes, ER-α and ER-β. Characterization assays used to profile new estrogen receptor ligands often utilize receptors from different species, with the assumption that they behave identically. To test this belief, we have profiled a no. of estrogens, other steroids, phytoestrogens and selective estrogen receptor modulators in a solid phase radioligand binding assay using recombinant protein for human, rat, and mouse ER-α and ER-β. Certain compds. show species dependent binding preferences for ER-α or ER-β, leading to differences in receptor subtype selectivity. The amino acids identified by crystallog. as lining the ligand binding cavity are the same among the three species, suggesting that as yet unidentified amino acids contribute to the structure of the binding site. We conclude from this anal. that the ability of a compd. to selectively bind to a particular ER subtype can be species dependent.
- 24Mohler, M. L.; Narayanan, R.; Coss, C. C.; Hu, K.; He, Y.; Wu, Z.; Hong, S.-S.; Hwang, D. J.; Miller, D. D.; Dalton, J. T. Estrogen Receptor β Selective Nonsteroidal Estrogens: Seeking Clinical Indications. Expert Opin. Ther. Pat. 2010, 20, 507– 534, DOI: 10.1517/13543771003657164Google Scholar24https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXjslamu7w%253D&md5=43641194b70be56c763fdf4d8e836e88Estrogen receptor β selective nonsteroidal estrogens: seeking clinical indicationsMohler, Michael L.; Narayanan, Ramesh; Coss, Christopher C.; Hu, Kejiang; He, Yali; Wu, Zhongzhi; Hong, Seoung-Soo; Hwang, Dong Jin; Miller, Duane D.; Dalton, James T.Expert Opinion on Therapeutic Patents (2010), 20 (4), 507-534CODEN: EOTPEG; ISSN:1354-3776. (Informa Healthcare)A review. Importance of the field: Nonsteroidal estrogens have been known since the 1930s. However, the relatively recent (1996) discovery of estrogen receptor subtype β (ERβ) suggested a possible paradigm shift away from SERM-like selectivity. Selective ERβ agonism would potentially allow expansion of estrogenic targeting into new indications (discussed herein) currently precluded by the thrombogenic and hyperproliferative effects of nonselective estrogens. Areas covered in this review: ERβ agonist design has been very successful. Pharmacophores for ERβ selective nonsteroidal estrogens are generally diphenolic compds. that achieve an inter-phenolic distance and geometry similar to 17β-estradiol with few restraints on the nature of the element linking the phenols (or phenol mimetics). The tremendously chemodiverse ERβ agonist patent literature is reviewed, segregating the agonists into structurally similar compds. based on their interphenolic linking elements. What the reader will gain: A comprehensive understanding of the chemotype landscape of this field and an assessment of its maturation. Take home message: Subtype selective ERβ agonist therapy seems very promising. However, more clin. testing is needed to firmly establish its therapeutic potential. At this point, ERβ is a promising target in search of an indication.
- 25Malamas, M. S.; Manas, E. S.; McDevitt, R. E.; Gunawan, I.; Xu, Z. B.; Collini, M. D.; Miller, C. P.; Dinh, T.; Henderson, R. A.; Keith, J. C.; Harris, H. A. Design and Synthesis of Aryl Diphenolic Azoles as Potent and Selective Estrogen Receptor-β Ligands. J. Med. Chem. 2004, 47, 5021– 5040, DOI: 10.1021/jm049719yGoogle Scholar25https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXnt12gtrc%253D&md5=01c7d1d0fb71105c1adffeb5f0bc7d3bDesign and Synthesis of Aryl Diphenolic Azoles as Potent and Selective Estrogen Receptor-β LigandsMalamas, Michael S.; Manas, Eric S.; McDevitt, Robert E.; Gunawan, Iwan; Xu, Zhang B.; Collini, Michael D.; Miller, Chris P.; Dinh, Tam; Henderson, Ruth A.; Keith, James C., Jr.; Harris, Heather A.Journal of Medicinal Chemistry (2004), 47 (21), 5021-5040CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)New diphenolic azoles as highly selective estrogen receptor-β agonists are reported. The more potent and selective analogs of these series have comparable binding affinities for ERβ as the natural ligand 17β-estradiol but are >100-fold selective over ERα. The design strategy not only followed a traditional SAR approach but also was supported by X-ray structures of ERβ cocrystd. with various ligands as well as mol. modeling studies. These strategies enabled the use of a single conservative residue substitution in the ligand-binding pocket, ERα Met421 → ERβ Ile373, to optimize ERβ selectivity. The 7-position-substituted benzoxazoles were the most selective ligands of both azole series, with ERB-041 (I) being >200-fold selective for ERβ. The majority of ERβ selective agonists tested that were at least ∼50-fold selective displayed a consistent in vivo profile: they were inactive in several models of classic estrogen action (uterotrophic, osteopenia, and vasomotor instability models) and yet were active in the HLA-B27 transgenic rat model of inflammatory bowel disease. These data suggest that ERβ-selective agonists are devoid of classic estrogenic effects and may offer a novel therapy to treat certain inflammatory conditions.
- 26Roman-Blas, J. A.; Castañeda, S.; Cutolo, M.; Herrero-Beaumont, G. Efficacy and Safety of a Selective Estrogen Receptor β Agonist, ERB-041, in Patients with Rheumatoid Arthritis: A 12-Week, Randomized, Placebo-Controlled, Phase II Study. Arthritis Care Res. 2010, 62, 1588– 1593, DOI: 10.1002/acr.20275Google Scholar26https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXhsF2gu7rF&md5=b3b3d9966e0c841340d2077af7cb43e7Efficacy and safety of a selective estrogen receptor β agonist, ERB-041, in patients with rheumatoid arthritis: a 12-week, randomized, placebo-controlled, phase II studyRoman-Blas, Jorge A.; Castaneda, Santos; Cutolo, Maurizio; Herrero-Beaumont, GabrielArthritis Care & Research (2010), 62 (11), 1588-1593CODEN: ARCREG; ISSN:2151-464X. (John Wiley & Sons, Inc.)Objective. Selective estrogen receptor β (ERβ) agonists have demonstrated relevant antiinflammatory effects in different animal models. This study aimed to compare the efficacy and safety of one of these agonists, ERB-041, in subjects with rheumatoid arthritis (RA). Methods. A total of 291 patients with active RA receiving stable doses of methotrexate were randomized to receive 5, 25, or 75 mg of ERB-041 or placebo for 12 wk. The primary end point was the American College of Rheumatol. 20% improvement criteria (ACR20) at 12 wk. Secondary end points included the ACR 50% improvement criteria (ACR50) and the ACR 70% improvement criteria (ACR70) responses, health outcomes measures, C-reactive protein (CRP) levels, and potential exposure-response relationships. Medical history, phys. examn., and lab. values were obtained at screening, baseline, and weeks 2, 4, 8, and 12. Results. No statistically significant difference for the ACR20 was found between the ERB-041 treatment and placebo groups (P = 0.518). Nor was a significant difference obsd. for ACR50 and ACR70 responses, health outcomes measures, CRP levels, and overall incidence of adverse events among all groups. Forty-four subjects (15.1%) discontinued the study and the rate of discontinuation was similar among the treatment groups. The most commonly reported treatment-emergent adverse events were headache (7.6%), nausea (6.2%), infection (4.8%), and bronchitis (4.1%). None of the adverse events was considered treatment related. Conclusion. Although well tolerated and safe, ERB-041 failed to demonstrate antiinflammatory efficacy in RA patients, despite evidence of strong activity in preclin. arthritis models. These results suggest that selective ERβ agonists would not have effects on regulating inflammatory response in RA. Nevertheless, further studies are warranted to establish their efficacy in inflammatory arthritis.
- 27Norman, B. H.; Dodge, J. A.; Richardson, T. I.; Borromeo, P. S.; Lugar, C. W.; Jones, S. A.; Chen, K.; Wang, Y.; Durst, G. L.; Barr, R. J.; Montrose-Rafizadeh, C.; Osborne, H. E.; Amos, R. M.; Guo, S.; Boodhoo, A.; Krishnan, V. Benzopyrans Are Selective Estrogen Receptor β Agonists with Novel Activity in Models of Benign Prostatic Hyperplasia. J. Med. Chem. 2006, 49, 6155– 6157, DOI: 10.1021/jm060491jGoogle Scholar27https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD28XptFaitr8%253D&md5=8d57f3742a46fe2b1ff176186a6f5d08Benzopyrans Are Selective Estrogen Receptor β Agonists with Novel Activity in Models of Benign Prostatic HyperplasiaNorman, Bryan H.; Dodge, Jeffrey A.; Richardson, Timothy I.; Borromeo, Peter S.; Lugar, Charles W.; Jones, Scott A.; Chen, Keyue; Wang, Yong; Durst, Gregory L.; Barr, Robert J.; Montrose-Rafizadeh, Chahrzad; Osborne, Harold E.; Amos, Robert M.; Guo, Sherry; Boodhoo, Amechand; Krishnan, VenkateshJournal of Medicinal Chemistry (2006), 49 (21), 6155-6157CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)Benzopyran selective estrogen receptor beta agonist-1 (SERBA-1) shows potent, selective binding and agonist function in estrogen receptor β (ERβ) in vitro assays. X-ray crystal structures of SERBA-1 in ERα and β help explain obsd. β-selectivity of this ligand. SERBA-1 in vivo demonstrates involution of the ventral prostate in CD-1 mice (ERβ effect), while having no effect on gonadal hormone levels (ERα effect) at 10× the efficacious dose, consistent with in vitro properties of this mol.
- 28Roehrborn, C. G.; Spann, M. E.; Myers, S. L.; Serviss, C. R.; Hu, L.; Jin, Y. Estrogen Receptor Beta Agonist LY500307 Fails to Improve Symptoms in Men with Enlarged Prostate Secondary to Benign Prostatic Hypertrophy. Prostate Cancer Prostatic Dis. 2015, 18, 43– 48, DOI: 10.1038/pcan.2014.43Google Scholar28https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXhvVOnt73I&md5=a6fa7058d61630f48df3b35c5d637c70Estrogen receptor beta agonist LY500307 fails to improve symptoms in men with enlarged prostate secondary to benign prostatic hypertrophyRoehrborn, C. G.; Spann, M. E.; Myers, S. L.; Serviss, C. R.; Hu, L.; Jin, Y.Prostate Cancer and Prostatic Diseases (2015), 18 (1), 43-48CODEN: PCPDFW; ISSN:1365-7852. (Nature Publishing Group)Background: To assess the efficacy and safety of LY500307, a selective estrogen receptor beta agonist, on lower urinary tract symptoms (LUTS) in patients with enlarged prostate secondary to BPH. Methods: In a randomized, double-blind, placebo-controlled, parallel phase 2, efficacy and safety study, eligible patients with moderate to severe LUTS and prostatic enlargement (≥30 mL) were randomized to placebo or LY500307 at 1, 3, 10 and 25 mg once daily for 24 wk. Primary efficacy end point was change in total International Prostate Symptoms Score (IPSS) after 24 wk. Secondary end points included changes in total prostate vol. (TPV) that served as a proof of concept end point, as well as IPSS quality of life, max. peak urine flow rate (Qmax) and PSA and safety (adverse events, lab. test). Results: A total of 414 patients were randomized when the study was terminated because of insufficient TPV redn., based on a priori defined interim anal. The IPSS mean change from baseline to end point was -3.4±6.8 in the placebo group and -1.3±6.6, -2.6±7.0, -3.7±6.7 and -4.4±5.7 in the 1, 3, 10 and 25 mg LY500307-treated groups, resp. (P>0.05). Similarly, no treatment effect was obsd. for any of the secondary efficacy measures. Incidence of adverse events was comparable between treatment groups, and no clin. meaningful changes in lab. tests were obsd. Conclusions: LY500307 was well tolerated in BPH patients with LUTS at doses up to 25 mg once daily for 24 wk. The study was terminated early because of inadequate efficacy.
- 29Breier, A. The Efficacy and Safety of a Selective Estrogen Receptor Beta Agonist (LY500307) for Negative Symptoms and Cognitive Impairment Associated With Schizophrenia—Full Text View—ClinicalTrials.gov. https://clinicaltrials.gov/ct2/show/NCT01874756 (accessed Apr 27, 2020).Google ScholarThere is no corresponding record for this reference.
- 30Effects of an ER Beta Agonist (Lilly Compound LY500307) on Estradiol-Withdrawal-Induced Mood Symptoms in Women with Past Perimenopausal Depression—Tabular View—ClinicalTrials.gov. https://clinicaltrials.gov/ct2/show/record/NCT03689543 (accessed Apr 27, 2020).Google ScholarThere is no corresponding record for this reference.
- 31Paterni, I.; Bertini, S.; Granchi, C.; Macchia, M.; Minutolo, F. Estrogen Receptor Ligands: A Patent Review Update. Expert Opin. Ther. Pat. 2013, 23, 1247– 1271, DOI: 10.1517/13543776.2013.805206Google Scholar31https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhsVyns7fO&md5=6f747f7fccf0953fc64cd61b5f287dd6Estrogen receptor ligands: a patent review updatePaterni, Ilaria; Bertini, Simone; Granchi, Carlotta; Macchia, Marco; Minutolo, FilippoExpert Opinion on Therapeutic Patents (2013), 23 (10), 1247-1271CODEN: EOTPEG; ISSN:1354-3776. (Informa Healthcare)A review. Introduction: The role of estrogens is mostly mediated by two nuclear receptors (ERα and ERβ) and a membrane-assocd. G-protein (GPR30 or GPER), and it is not limited to reprodn., but it extends to the skeletal, cardiovascular and central nervous systems. Various pathologies such as cancer, inflammatory, neurodegenerative and metabolic diseases are often assocd. with dysfunctions of the estrogenic system. Therapeutic interventions by agents that affect the estrogenic signaling pathway might be useful in the treatment of many dissimilar diseases. Areas covered: The massive chemodiversity of ER ligands, limited to patented small mols., is herein reviewed. The reported compds. are classified on the basis of their chem. structures. Non-steroidal derivs., which mostly consist of diphenolic compds., are further segregated into chem. classes based on their central scaffold. Expert opinion: Estrogens have been used for almost a century and their earlier applications have concerned interventions in the female reproductive functions, as well as the treatment of some estrogen-dependent cancers and osteoporosis. Since the discovery of ERβ in 1996, the patent literature has started to pay a progressively increasing attention to this newer receptor subtype, which holds promise as a target for new indications, most of which still need to be clin. validated.
- 32Endo, Y.; Iijima, T.; Yamakoshi, Y.; Yamaguchi, M.; Fukasawa, H.; Shudo, K. Potent Estrogenic Agonists Bearing Dicarba-closo-Dodecaborane as a Hydrophobic Pharmacophore. J. Med. Chem. 1999, 42, 1501– 1504, DOI: 10.1021/jm9900725Google Scholar32https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXisVaqsbY%253D&md5=b487b1fcfa55e50d83a5cf96d8b357faPotent Estrogenic Agonists Bearing Dicarba-closo-dodecaborane as a Hydrophobic PharmacophoreEndo, Yasuyuki; Iijima, Toru; Yamakoshi, Yuko; Yamaguchi, Mitsuhiro; Fukasawa, Hiroshi; Shudo, KoichiJournal of Medicinal Chemistry (1999), 42 (9), 1501-1504CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)The novel carborane contg. mols., e.g. 1-(4-hydroxyphenyl)-1,12-dicarba-closo-dodecaborane, with potent estrogenic activity was developed. The unique character of biol. active mols. contg. a carborane skeleton may give to unusual membrane transport characteristics and metab., compared with conventional active mols. The superagonistic properties of the carborane-contg. compds. raise the possibility that structure-function studies could lead to the development of more selective estrogen agonists and antagonists, which could be useful as therapeutic agents for a wide variety of conditions.
- 33Barth, R. F.; Zhang, Z.; Liu, T. A Realistic Appraisal of Boron Neutron Capture Therapy as a Cancer Treatment Modality. Cancer Commun. 2018, 38, 36 DOI: 10.1186/s40880-018-0280-5Google Scholar33https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC1MbosFamuw%253D%253D&md5=aaf7960e10efcae0cbdcaa5113ba2bdaA realistic appraisal of boron neutron capture therapy as a cancer treatment modalityBarth Rolf F; Zhang Zizhu; Liu TongCancer communications (London, England) (2018), 38 (1), 36 ISSN:.Boron neutron capture therapy (BNCT) is a binary therapeutic modality based on the nuclear capture and fission reactions that occur when the stable isotope boron-10 is irradiated with neutrons to produce high-energy alpha particles and recoiling lithium-7 nuclei. In this Commentary we will focus on a number of papers that were presented at a Symposium entitled "Current Clinical Status of Boron Neutron Capture Therapy and Paths to the Future", which was held in September 2017 at the China National Convention Center in Beijing. Results were presented by clinicians from Japan, Finland, the United States, the China mainland and Taiwan, China who have been working in the multiple disciplines that are required for carrying out clinical BNCT. The main focus was on the treatment of patients with malignant brain tumors, recurrent tumors of the head and neck region, and cutaneous melanomas. The results obtained in treating these patients were reported in detail and, although most of the patients with brain tumors and head and neck cancer were not cured, there was evidence of some clinical efficacy. Although there are a number of problems that must be addressed, further clinical studies to evaluate the efficacy of BNCT are warranted. First, despite considerable effort by numerous investigators over the past 40 years, there still are only two boron-containing drugs in clinical use, L-boronophenylalanine (BPA) and sodium borocaptate (BSH). Therefore, until new and more effective boron delivery agents are developed, efforts should be directed to improving the dosing and delivery of BPA and BSH. Second, due to a variety of reasons, nuclear reactor-based BNCT has ended except for its use in the China mainland and Taiwan. Therefore, the future of BNCT depends upon the results of the ongoing Phase II clinical trials that are being carried out in Japan and the soon to be initiated trials that will be carried out in Finland. If the results obtained from these clinical trials are sufficiently promising, then BNCT will have a clear path to the future, especially for patients with the therapeutically challenging malignancies that in the past have been treated with reactor-based BNCT.
- 34Soloway, A. H.; Tjarks, W.; Barnum, B. A.; Rong, F.-G.; Barth, R. F.; Codogni, I. M.; Wilson, J. G. The Chemistry of Neutron Capture Therapy. Chem. Rev. 1998, 98, 1515– 1562, DOI: 10.1021/cr941195uGoogle Scholar34https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1cXjtlygtL8%253D&md5=52bcd4d4e2f37833d76092e9a1867b3cThe Chemistry of Neutron Capture TherapySoloway, Albert H.; Tjarks, Werner; Barnum, Beverly A.; Rong, Feng-Guang; Barth, Rolf F.; Codogni, Iwona M.; Wilson, J. GeraldChemical Reviews (Washington, D. C.) (1998), 98 (4), 1515-1562CODEN: CHREAY; ISSN:0009-2665. (American Chemical Society)A review with 633 refs.
- 35Soloway, A. H.; Wright, R. L.; Messer, J. R. Evaluation of Boron Compounds for Use in Neutron Capture Therapy of Brain Tumors. I. Animal Investigations. J. Pharmacol. Exp. Ther. 1961, 134, 117– 122Google Scholar35https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaF38Xks1al&md5=e291c2430bd5f235a4b0f6e3a429a7d3Evaluation of boron compounds for use in neutron-capture therapy of brain tumors. I. Animal investigationsSoloway, A. H.; Wright, R. L.; Messer, J. R.Journal of Pharmacology and Experimental Therapeutics (1961), 134 (), 117-22CODEN: JPETAB; ISSN:0022-3565.cf. CA 54, 1374d.-m-Boronosuccinanilic, 3-amino-4-carboxybenzeneboronic, m- and p-carboxybenzeneboronic, 2-nitrobenzene-l,4-diboronic, o-(2-carboxyacetamidoethyl)benzeneboronic, 2-acetamidobenzeneboronic, and m-ureidobenzeneboronic acids, p-boronophenylalanine, Na perhydrodecaborate, and boric acid were evaluated in mice with transplanted glioma (ependymoma) for toxicity. The tumor-to-brain ratios and the concns. in various organ tissues were detd. The 1st 2 compds. appeared promising and were studied further in cats by intravenous and intracarotid artery injection.
- 36Stockmann, P.; Gozzi, M.; Kuhnert, R.; Sárosi, M. B.; Hey-Hawkins, E. New Keys for Old Locks: Carborane-Containing Drugs as Platforms for Mechanism-Based Therapies. Chem. Soc. Rev. 2019, 48, 3497– 3512, DOI: 10.1039/C9CS00197BGoogle Scholar36https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhtF2nt7vI&md5=b2a1f1247161aeedf257cd807d2a30fdNew keys for old locks: carborane-containing drugs as platforms for mechanism-based therapiesStockmann, Philipp; Gozzi, Marta; Kuhnert, Robert; Sarosi, Menyhart B.; Hey-Hawkins, EvamarieChemical Society Reviews (2019), 48 (13), 3497-3512CODEN: CSRVBR; ISSN:0306-0012. (Royal Society of Chemistry)A review. Icosahedral carboranes in medicine are still an emerging class of compds. with potential beneficial applications in drug design. These highly hydrophobic clusters are potential "new keys for old locks" which open up an exciting field of research for well-known, but challenging important therapeutic substrates, as demonstrated by the numerous examples discussed in this review.
- 37Leśnikowski, Z. J. Challenges and Opportunities for the Application of Boron Clusters in Drug Design. J. Med. Chem. 2016, 59, 7738– 7758, DOI: 10.1021/acs.jmedchem.5b01932Google Scholar37https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XmvFSgs7c%253D&md5=1d4c3720cdc649c5fc3de397bea15e83Challenges and Opportunities for the Application of Boron Clusters in Drug DesignLesnikowski, Zbigniew J.Journal of Medicinal Chemistry (2016), 59 (17), 7738-7758CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)There are two branches in boron medicinal chem.: the first focuses on single boron atom compds., and the second utilizes boron clusters. Boron clusters and their heteroatom counterparts belong to the family of cage compds. A subset of this extensive class of compds. includes dicarbadodecaboranes, which have the general formula C2B10H12, and their metal biscarboranyl complexes, metallacarboranes, with the formula [M(C2B10H12)2-2]. The unique properties of boron clusters have resulted in their utilization in applications such as in pharmacophores, as scaffolds in mol. construction, and as modulators of bioactive compds. This Perspective presents an overview of the properties of boron clusters that are pertinent for drug discovery, recent applications in the design of various classes of drugs, and the potential use of boron clusters in the construction of new pharmaceuticals.
- 38Leśnikowski, Z. J. What Are the Current Challenges with the Application of Boron Clusters to Drug Design?. Expert Opin. Drug Discovery 2020, 481, DOI: 10.1080/17460441.2021.1867531Google ScholarThere is no corresponding record for this reference.
- 39Sedlák, D.; Eignerová, B.; Dračínský, M.; Janoušek, Z.; Bartůněk, P.; Kotora, M. Synthesis and Evaluation of 17α-(Carboranylalkyl)Estradiols as Ligands for Estrogen Receptors α and β. J. Organomet. Chem. 2013, 747, 178– 183, DOI: 10.1016/j.jorganchem.2013.06.013Google Scholar39https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhtVKgsrrL&md5=30ff55174dd93879e3cdd196e1032c6aSynthesis and evaluation of 17α-(carboranylalkyl)estradiols as ligands for estrogen receptors α and βSedlak, David; Eignerova, Barbara; Dracinsky, Martin; Janousek, Zbynek; Bartunek, Petr; Kotora, MartinJournal of Organometallic Chemistry (2013), 747 (), 178-183CODEN: JORCAI; ISSN:0022-328X. (Elsevier B.V.)A series of 17α-(carboranylalkyl)estradiols was synthesized using cross-metathesis reaction of 17α-allyl- and 17α-vinylestradiols with allylcarboranes catalyzed by Hoveyda-Grubbs 2nd generation catalyst. The prepd. estradiol derivs. were tested in the panel of cell-based reporter assays including all steroid receptors. The compds. mainly activated estrogen receptors α and β and tended to be weakly selective for ERα. Besides ERα and ERβ, we have also detected a weak agonistic activity on AR and PR at micromolar concns. We didn't observe any antagonistic effect with the exception of two receptors: GR and MR which were inhibited with some of the tested ligands. However, the inhibition was detectable at concns. exceeding by far those needed to activate estrogen receptors.
- 40Eignerová, B.; Sedlák, D.; Dračínský, M.; Bartůněk, P.; Kotora, M. Synthesis and Biochemical Characterization of a Series of 17α-Perfluoroalkylated Estradiols as Selective Ligands for Estrogen Receptor α. J. Med. Chem. 2010, 53, 6947– 6953, DOI: 10.1021/jm100563hGoogle Scholar40https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXhtFWlsb7E&md5=be45ac8fd4423d38ba410b2e080676faSynthesis and Biochemical Characterization of a Series of 17α-Perfluoroalkylated Estradiols as Selective Ligands for Estrogen Receptor αEignerova, Barbara; Sedlak, David; Dracinsky, Martin; Bartunek, Petr; Kotora, MartinJournal of Medicinal Chemistry (2010), 53 (19), 6947-6953CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)Despite intensive research efforts, the distinct biol. roles of two closely related estrogen receptors, ERα and ERβ, are only partially understood. Therefore, ligands selective for either of two isotypes are useful research tools because they allow for exerting a desired subset of biol. effects mediated by only one of the receptors. Here we report on the synthesis of a new class of potent and selective ligands for ERα represented by a series of 17α-substituted estradiols bearing lipophilic perfluoroalkyl chains. These 17α-perfluoroalkylated estradiols were synthesized by Ru-catalyzed cross metathesis reactions of 17α-allyl- or 17α-vinylestradiols with perfluoroalkylpropenes. Compds. were tested in both agonistic and antagonistic modes using a panel of stable steroid receptor reporter cell lines established in U2OS cells and consisting of ERα-LBD, ERβ-LBD, GR-LBD, and MR-LBD reporters. Some of the compds. are potent and selective agonists of ERα, exhibiting weak partial to no detectable agonistic activity on ERβ. Notably, I is the most ERα selective ligand of the prepd. compds. because it activates ERα but inhibits ERβ. In addn., some compds. are pure agonists on ERα but show mixed agonistic/antagonistic profile on ERβ which is a typical pattern obsd. for selective estrogen receptor modulators (SERMs).
- 41Sedlák, D.; Novák, P.; Kotora, M.; Bartůněk, P. Synthesis and Evaluation of 17α-Arylestradiols as Ligands for Estrogen Receptor α and β. J. Med. Chem. 2010, 53, 4290– 4294, DOI: 10.1021/jm901898aGoogle Scholar41https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXltVyhuro%253D&md5=f1c7acaa44d748d6c5adc341290a79cbSynthesis and Evaluation of 17α-Arylestradiols as Ligands for Estrogen Receptor α and βSedlak, David; Novak, Petr; Kotora, Martin; Bartunek, PetrJournal of Medicinal Chemistry (2010), 53 (10), 4290-4294CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)To identify novel estrogen receptor ligands a series of substituted 17α-arylestradiols I [X = C(CO2Et)2, C(COMe)2, C(CO2Et)COMe, C(CO2Et)CN, NTs, O] were synthesized using the catalytic [2 + 2 + 2]-cyclotrimerization of 17α-ethynylestradiol with various 1,7-diynes HC≡CCH2XCH2C≡CH in the presence of Wilkinson's catalysts [Rh(PPh3)3Cl]. The compds. were subjected to competitive binding assays, cell-based luciferase reporter assays, and proliferation assays. These expts. confirmed their estrogenic character and revealed some interesting properties like mixed partial/full agonism for ERα/ERβ and different selectivity as a result of differing potencies for either ER.
- 42Endo, Y.; Yamamoto, K.; Kagechika, H. Utility of Boron Clusters for Drug Design. Relation between Estrogen Receptor Binding Affinity and Hydrophobicity of Phenols Bearing Various Types of Carboranyl Groups. Bioorg. Med. Chem. Lett. 2003, 13, 4089– 4092, DOI: 10.1016/j.bmcl.2003.08.039Google Scholar42https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3sXosVWrtrk%253D&md5=337630c26a43eb82c590ebb7b6cd8e15Utility of boron clusters for drug design. Relation between estrogen receptor binding affinity and hydrophobicity of phenols bearing various types of carboranyl groupsEndo, Yasuyuki; Yamamoto, Keisuke; Kagechika, HiroyukiBioorganic & Medicinal Chemistry Letters (2003), 13 (22), 4089-4092CODEN: BMCLE8; ISSN:0960-894X. (Elsevier Science B.V.)High binding affinity for estrogen receptor and the appearance of estrogenic activity require a phenolic ring and an appropriate hydrophobic group adjacent to the phenolic ring. A quant. structure-activity relationship anal. based on the values of logP and the pKa of the phenolic group showed that the hydrophobicity of these compds. is highly correlated to the estrogen receptor α (ERα)-binding affinity. These results should be useful for application of these spherical boron clusters (dicarba-closo-dodecaboranes; carboranes) as hydrophobic pharmacophores in drug design, as well as for microscopic anal. of ER-ligand interactions.
- 43Endo, Y.; Iijima, T.; Yamakoshi, Y.; Fukasawa, H.; Miyaura, C.; Inada, M.; Kubo, A.; Itai, A. Potent Estrogen Agonists Based on Carborane as a Hydrophobic Skeletal Structure: A New Medicinal Application of Boron Clusters. Chem. Biol. 2001, 8, 341– 355, DOI: 10.1016/S1074-5521(01)00016-3Google Scholar43https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3MXjs1ahsr4%253D&md5=1f1af2e8526772e69d711f538fc1d82dPotent estrogen agonists based on carborane as a hydrophobic skeletal structure: a new medicinal application of boron clustersEndo, Yasuyuki; Iijima, Toru; Yamakoshi, Yuko; Fukasawa, Hiroshi; Miyaura, Chisato; Inada, Masaki; Kubo, Asako; Itai, AkikoChemistry & Biology (2001), 8 (4), 341-355CODEN: CBOLE2; ISSN:1074-5521. (Elsevier Science Ltd.)Background: Carboranes (dicarba-closo-dodecaboranes) are a class of carbon-contg. polyhedral boron-cluster compds. having remarkable thermal stability and exceptional hydrophobicity. Applications of the unique structural and chem. properties offered by icosahedral carboranes in boron neutron capture therapy have received increasing attention over the past 30 yr. However, these features of carboranes may allow another application as a hydrophobic pharmacophore in biol. active mols. that interact hydrophobically with receptors. Results: We have designed candidate estrogen-receptor-binding compds. having carborane as a hydrophobic skeletal structure and synthesized them. The most potent compd. bearing a carborane cage exhibited activity at least 10-fold greater than that of 17β-estradiol in the luciferase reporter gene assay. Estrogen receptor-α-binding data for the compd. were consistent with the results of the luciferase reporter gene assay. The compd. also showed potent in vivo effects on the recovery of uterine wt. and bone loss in ovariectomized mice. Conclusion: Further development of the potent carborane-contg. estrogenic agonists described here, having a new skeletal structure and unique characteristics, should yield novel therapeutic agents, esp. selective estrogen receptor modulators. Furthermore, the suitability of the spherical carborane cage for binding to the cavity of the estrogen receptor-α ligand-binding domain should provide a basis for a similar approach to developing novel ligands for other steroid receptors.
- 44Ohta, K.; Ogawa, T.; Oda, A.; Kaise, A.; Endo, Y. Design and Synthesis of Carborane-Containing Estrogen Receptor-Beta (ERβ)-Selective Ligands. Bioorg. Med. Chem. Lett. 2015, 25, 4174– 4178, DOI: 10.1016/j.bmcl.2015.08.007Google Scholar44https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2MXhtlSlsLjF&md5=18ce9668981a0f26829a981d79f8addbDesign and synthesis of carborane-containing estrogen receptor-beta (ERβ)-selective ligandsOhta, Kiminori; Ogawa, Takumi; Oda, Akifumi; Kaise, Asako; Endo, YasuyukiBioorganic & Medicinal Chemistry Letters (2015), 25 (19), 4174-4178CODEN: BMCLE8; ISSN:0960-894X. (Elsevier B.V.)Candidates for highly selective estrogen receptor-beta (ERβ) ligands were designed and synthesized based on carborane-contg. ER ligands as lead compds. Among them, p-carboranylcyclohexanol derivs. 8a and 8b exhibited high ERβ selectivity in competitive binding assay: for example, 8a showed 56-fold selectivity for ERβ over ERα. Docking studies of 8a and 8b with the ERα and ERβ ligand-binding domains (LBDs) suggested that the p-carborane cage of the ligands is located close to key amino acid residues that influence ER-subtype selectivity, i.e., Leu384 in the ERα LBD and Met336 in the ERβ LBD. The p-carborane cage in 8a and 8b appears to play a crucial role in the increased ERβ selectivity.
- 45Coult, R.; Fox, M. A.; Gill, W. R.; Herbertson, P. L.; MacBride, J. A. H.; Wade, K. C-Arylation and C-Heteroarylation of Icosahedral Carboranes via Their Copper(I) Derivatives. J. Organomet. Chem. 1993, 462, 19– 29, DOI: 10.1016/0022-328X(93)83337-UGoogle Scholar45https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2cXitFOktbc%253D&md5=cbd713095584fc4dbaf5b88cee325275C-arylation and C-heteroarylation of icosahedral carboranes via their copper(I) derivativesCoult, Robert; Fox, Mark A.; Gill, Wendy R.; Herbertson, Penelope L.; MacBride, J. A. Hugh; Wade, KennethJournal of Organometallic Chemistry (1993), 462 (1-2), 19-29CODEN: JORCAI; ISSN:0022-328X.Reaction between C-mono- or C,C'-di-copper(I) derivs. of 1,2-, 1,7-, or 1,12-dicarba-closo-dodecaborane(12) and aryl iodides in the presence of pyridine gives the corresponding C-mono- or C,C'-diaryl derivs. of 1,7- and 1,12-dicarba-closo-dodecaboranes(12); 1,2-dicarba-closo-dodecaborane(12) gives only the C-monoaryl product. Cyclic or linear arylene coupled systems are obtained when di-iodoarenes are used. Copper(I) derivs. may be generated from C-unsubstituted or C-monosubstituted carboranes using copper(I) t-butoxide when substituents incompatible with the use of C-lithio-intermediates are involved. The C-copper(I) deriv. of 1,2-dicarba-closo-dodecaborane(12) gives 1,2-di-2'-pyridyl-1,2-dicarba-closo-dodecaborane(12) specifically with 2-bromopyridine. The (inferred) intermediate mono-2-pyridyl-deriv., obtained independently from 2-ethynylpyridine and the di-Me sulfide complex of decaborane, gives 1-phenyl-2,2'-pyridyl-1,2-dicarba-closo-dodecaborane(12) upon conversion into its copper(I) deriv. and treatment with iododobenzene. However, the copper(I) deriv. of 1-phenyl-1,2-dicarba-closo-dodecaborane(12) does not react to a significant extent with 2-bromopyridine.
- 46Corey, E. J.; Bakshi, R. K.; Shibata, S.; Chen, C. P.; Singh, V. K. A Stable and Easily Prepared Catalyst for the Enantioselective Reduction of Ketones. Applications to Multistep Syntheses. J. Am. Chem. Soc. 1987, 109, 7925– 7926, DOI: 10.1021/ja00259a075Google Scholar46https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaL1cXnsFShtA%253D%253D&md5=4deaae8502b4e75f0744e46a9e774decA stable and easily prepared catalyst for the enantioselective reduction of ketones. Applications to multistep synthesesCorey, E. J.; Bakshi, Raman K.; Shibata, Saizo; Chen, Chung Pin; Singh, Vinod K.Journal of the American Chemical Society (1987), 109 (25), 7925-6CODEN: JACSAT; ISSN:0002-7863.The readily available and stable oxazaborolidine I is an excellent catalyst for the enantioselective redn. of a wide range of ketones. The stereochem. course of the redn. is predictable and the secondary alcs. formed are generally produced with high enantioselectivity. Use of I or its enantiomer provides outstanding control of stereochem. at C(15) in prostaglandin synthesis. A general and highly enantioselective synthesis of trans-2,5-diarylfurans, potent antagonists of platelet activating factor, also illustrates the value of this enantioselective redn. Both I and its enantiomer can be obtained from (±)-pyroglutamic acid.
- 47Dale, J. A.; Mosher, H. S. Nuclear Magnetic Resonance Enantiomer Regents. Configurational Correlations via Nuclear Magnetic Resonance Chemical Shifts of Diastereomeric Mandelate, O-Methylmandelate, and.Alpha.-Methoxy-.Alpha.-Trifluoromethylphenylacetate (MTPA) Esters. J. Am. Chem. Soc. 1973, 95, 512– 519, DOI: 10.1021/ja00783a034Google Scholar47https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaE3sXotlOjtw%253D%253D&md5=27a7f9c81b89c6b29ffed705dc4f689cNuclear magnetic resonance enantiomer regents. Configurational correlations via nuclear magnetic resonance chemical shifts of diastereomeric mandelate, O-methylmandelate, and α-methoxy-α-trifluoromethylphenylacetate (MTPA) estersDale, James A.; Mosher, Harry S.Journal of the American Chemical Society (1973), 95 (2), 512-19CODEN: JACSAT; ISSN:0002-7863.An empirically derived correlation of configuration and NMR chem. shifts for diastereomeric mandelate, O-methylmandelate and α-methoxy-α-trifluoromethylphenylacetate esters has been developed and rationalized in terms of useful models. These models have been successfully applied to over 40 examples. The correlations involve the relative chem. shifts of the proton resonances from the groups attached to the carbinyl carbon of these diastereomeric esters.
- 48Hoye, T. R.; Jeffrey, C. S.; Shao, F. Mosher Ester Analysis for the Determination of Absolute Configuration of Stereogenic (Chiral) Carbinol Carbons. Nat. Protoc. 2007, 2, 2451– 2458, DOI: 10.1038/nprot.2007.354Google Scholar48https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXhtFOksL3E&md5=741bb9fd8eb7c18c310196f298e5ea3fMosher ester analysis for the determination of absolute configuration of stereogenic (chiral) carbinol carbonsHoye, Thomas R.; Jeffrey, Christopher S.; Shao, FengNature Protocols (2007), 2 (10), 2451-2458CODEN: NPARDW; ISSN:1750-2799. (Nature Publishing Group)This protocol details the most commonly used NMR-based method for deducing the configuration of otherwise unknown stereogenic, secondary carbinol (alc.) centers (R1R2CHOH (or the analogous amines where OH is replaced by NH2)). This 'Mosher ester anal.' relies on the fact that the protons in diastereomeric α-methoxy-α-trifluoromethylphenylacetic acid (MTPA) esters (i.e., those derived from conjugation of the carbinol under interrogation with MTPA) display different arrays of chem. shifts (δs) in their 1H NMR spectra. The protocol consists of the following: (i) prepn. of each of the diastereomeric S- and R-MTPA esters and (ii) comparative (ΔδSR) anal. of the 1H NMR spectral data of these two esters. By analyzing the sign of the difference in chem. shifts for a no. of analogous pairs of protons (the set of ΔδSR values) in the diastereomeric esters (or amides), the abs. configuration of the original carbinol (or amino) stereocenter can be reliably deduced. A typical Mosher ester anal. requires approx. 4-6 h of active effort over a 1- to 2-d period.
- 49Goto, T.; Ohta, K.; Fujii, S.; Ohta, S.; Endo, Y. Design and Synthesis of Androgen Receptor Full Antagonists Bearing a P-Carborane Cage: Promising Ligands for Anti-Androgen Withdrawal Syndrome. J. Med. Chem. 2010, 53, 4917– 4926, DOI: 10.1021/jm100316fGoogle Scholar49https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXmvFaktbs%253D&md5=e1df86c4bdb95869f329aac6442419d7Design and Synthesis of Androgen Receptor Full Antagonists Bearing a p-Carborane Cage: Promising Ligands for Anti-Androgen Withdrawal SyndromeGoto, Tokuhito; Ohta, Kiminori; Fujii, Shinya; Ohta, Shigeru; Endo, YasuyukiJournal of Medicinal Chemistry (2010), 53 (13), 4917-4926CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)Pure androgen receptor (AR) full antagonists are candidates to treat anti-androgen refractory prostate cancers. We previously developed a carborane-contg. AR antagonist, 3-(12-hydroxymethyl-1,12-dicarba-closo-dodecaborane-1-yl)benzonitrile (BA341), which was more potent than hydroxyflutamide (4) but acted as an agonist toward LNCaP prostate cancer cells expressing T877A AR mutant. Here, we designed and synthesized novel AR full antagonists, 3-[12-(1-hydroxy-2-Ar-ethyl)-1,12-dicarba-closo-dodecaborane-1-yl]benzonitrile (e.g., Ar = 4-XC6H4O, X = F 7b, MeO, NO2, NHCOMe and Ar = 4-XC6H4S, X = H, F 8b, MeO, NHCOMe, CF3 and Ar = 4-XC6H4SO2) structurally based upon the clin. used AR full antagonist (R)-bicalutamide (5) to test our hypothesis that the carborane cage is suitable as a hydrophobic pharmacophore for AR ligands. 7A and 8b showed good biol. profiles in AR binding and transactivation assays and dose-dependently inhibited the testosterone-induced proliferation of LNCaP cells, as well as SC-3 cells. The IC50 values of compds. 7b and 8b were 3.8 × 10-7 and 4.2 × 10-7 M, resp. [5, 8.7 × 10-7 M]. Since compds. 7b and 8b did not show any agonistic activity in functional assays, they seem to be pure AR full antagonists and are therefore candidates for treatment of anti-androgen withdrawal syndrome.
- 50Ohta, K.; Goto, T.; Fujii, S.; Kawahata, M.; Oda, A.; Ohta, S.; Yamaguchi, K.; Hirono, S.; Endo, Y. Crystal Structure, Docking Study and Structure–Activity Relationship of Carborane-Containing Androgen Receptor Antagonist 3-(12-Hydroxymethyl-1,12-Dicarba-closo-Dodecaboran-1-Yl)Benzonitrile. Bioorg. Med. Chem. 2011, 19, 3540– 3548, DOI: 10.1016/j.bmc.2011.04.017Google Scholar50https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXmvFWmt7Y%253D&md5=ad316a904afbef54577cf99651706340Crystal structure, docking study and structure-activity relationship of carborane-containing androgen receptor antagonist 3-(12-hydroxymethyl-1,12-dicarba-closo-dodecarboran-1-yl)benzonitrileOhta, Kiminori; Goto, Tokuhito; Fujii, Shinya; Kawahata, Masatoshi; Oda, Akifumi; Ohta, Shigeru; Yamaguchi, Kentaro; Hirono, Shuichi; Endo, YasuyukiBioorganic & Medicinal Chemistry (2011), 19 (11), 3540-3548CODEN: BMECEP; ISSN:0968-0896. (Elsevier B.V.)A potent androgen receptor (AR) antagonist, 3-(12-hydroxymethyl-1,12 dicarba-closo-dodecarboran-1-yl)benzonitrile (3a, BA341), contains a p-carborane cage as a hydrophobic pharmacophore. We elucidated the structural properties of 3a by single-crystal x-ray diffraction anal. and conducted a docking study of 3a with hAR LBD. The cyano group of 3a formed hydrogen bonds with Gln711 and Arg752 and the hydroxymethyl group did so with Asn705 and Thr877 in hAR LBD. The bulky p-carborane cage was accommodated in the hydrophobic pocket of hAR LBD. To understand the structure-activity relation around the hydroxymethyl group of 3a, we designed, synthesized, and evaluated the biol. activities of various novel AR ligands. Since the biol. activities of carbonyl compds. 8a, 8b, and 8c were similar to or weaker than those of the parent hydroxyl compds. 3a, 7a, and 7b, it seems to be necessary to have not only a hydrogen bonding acceptor, but also a hydrogen bonding donor adjacent to the hydroxymethyl group of 3a for efficient interaction with hAR LBD.
- 51Anzenbacher, P.; Zanger, U. M., Eds. Metabolism of Drugs and Other Xenobiotics, 1st ed.; John Wiley & Sons, Ltd., 2012.Google ScholarThere is no corresponding record for this reference.
- 52Lee, A. J.; Cai, M. X.; Thomas, P. E.; Conney, A. H.; Zhu, B. T. Characterization of the Oxidative Metabolites of 17β-Estradiol and Estrone Formed by 15 Selectively Expressed Human Cytochrome P450 Isoforms. Endocrinology 2003, 144, 3382– 3398, DOI: 10.1210/en.2003-0192Google Scholar52https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3sXlslGjsb0%253D&md5=dc77411de262647d6fcf0c28c27d66e7Characterization of the oxidative metabolites of 17β-estradiol and estrone formed by 15 selectively expressed human cytochrome p450 isoformsLee, Anthony J.; Cai, May Xiaoxin; Thomas, Paul E.; Conney, Allan H.; Zhu, Bao TingEndocrinology (2003), 144 (8), 3382-3398CODEN: ENDOAO; ISSN:0013-7227. (Endocrine Society)We systematically characterized the oxidative metabolites of 17β-estradiol and estrone formed by 15 human cytochrome P 450 (CYP) isoforms. CYP1A1 had high activity for 17β-estradiol 2-hydroxylation, followed by 15α-, 6α-, 4-, and 7α-hydroxylation. However, when estrone was the substrate, CYP1A1 formed more 4-hydroxyestrone than 15α- or 6α-hydroxyestrone, with 2-hydroxyestrone as the major metabolite. CYP1A2 had the highest activity for the 2-hydroxylation of both 17β-estradiol and estrone, although it also had considerable activity for their 4-hydroxylation (9-13% of 2-hydroxylation). CYP1B1 mainly catalyzed the formation of catechol estrogens, with 4-hydroxyestrogens predominant. CYP2A6, 2B6, 2C8, 2C9, 2C19, and 2D6 each showed a varying degree of low catalytic activity for estrogen 2-hydroxylation, whereas CYP2C18 and CYP2E1 did not show any detectable estrogen-hydroxylating activity. CYP3A4 had strong activity for the formation of 2-hydroxyestradiol, followed by 4-hydroxyestradiol and an unknown polar metabolite, and small amts. of 16α- and 16β-hydroxyestrogens were also formed. The ratio of 4- to 2-hydroxylation of 17β-estradiol or estrone with CYP3A4 was 0.22 or 0.51, resp. CYP3A5 had similar catalytic activity for the formation of 2- and 4-hydroxyestrogens. Notably, CYP3A5 had an unusually high ratio of 4- to 2-hydroxylation of 17β-estradiol or estrone (0.53 or 1.26, resp.). CYP3A4 and 3A5 also catalyzed the formation of nonpolar estrogen metabolite peaks (chromatog. less polar than estrone). CYP3A7 had a distinct catalytic activity for the 16α-hydroxylation of estrone, but not 17β-estradiol. CYP4A11 had little catalytic activity for the metab. of 17β-estradiol and estrone. In conclusion, many human CYP isoforms are involved in the oxidative metab. of 17β-estradiol and estrone, with a varying degree of catalytic activity and distinct regioselectivity.
- 53Yano, J. K.; Wester, M. R.; Schoch, G. A.; Griffin, K. J.; Stout, C. D.; Johnson, E. F. The Structure of Human Microsomal Cytochrome P450 3A4 Determined by X-ray Crystallography to 2.05-Å Resolution. J. Biol. Chem. 2004, 279, 38091– 38094, DOI: 10.1074/jbc.C400293200Google Scholar53https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXnt1Oqs7c%253D&md5=59a8fa760069d365165ddeeb9e4081d4The Structure of Human Microsomal Cytochrome P450 3A4 Determined by X-ray Crystallography to 2.05-Å ResolutionYano, Jason K.; Wester, Michael R.; Schoch, Guillaume A.; Griffin, Keith J.; Stout, C. David; Johnson, Eric F.Journal of Biological Chemistry (2004), 279 (37), 38091-38094CODEN: JBCHA3; ISSN:0021-9258. (American Society for Biochemistry and Molecular Biology)The structure of P 450 3A4 was detd. by x-ray crystallog. to 2.05-Å resoln. P 450 3A4 catalyzes the metabolic clearance of a large no. of clin. used drugs, and a no. of adverse drug-drug interactions reflect the inhibition or induction of the enzyme. P 450 3A4 exhibits a relatively large substrate-binding cavity that is consistent with its capacity to oxidize bulky substrates such as cyclosporin, statins, taxanes, and macrolide antibiotics. Family 3A P450s also exhibit unusual kinetic characteristics that suggest simultaneous occupancy by smaller substrates. Although the active site vol. is similar to that of P 450 2C8 (PDB code: 1PQ2), the shape of the active site cavity differs considerably due to differences in the folding and packing of portions of the protein that form the cavity. Compared with P 450 2C8, the active site cavity of 3A4 is much larger near the heme iron. The lower constraints on the motions of small substrates near the site of oxygen activation may diminish the efficiency of substrate oxidn., which may in turn be improved by space restrictions imposed by the presence of a second substrate mol. The structure of P 450 3A4 should facilitate a better understanding of the substrate selectivity of the enzyme.
- 54Anzenbacherová, E.; Bec, N.; Anzenbacher, P.; Hudecek, J.; Soucek, P.; Jung, C.; Munro, A. W.; Lange, R. Flexibility and Stability of the Structure of Cytochromes P450 3A4 and BM-3. Eur. J. Biochem. 2000, 267, 2916– 2920, DOI: 10.1046/j.1432-1327.2000.01305.xGoogle Scholar54https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3cXjs1Gjs7w%253D&md5=9e358cae19770cf66d9ec8cd6ffe595eFlexibility and stability of the structure of cytochromes P450 3A4 and BM-3Anzenbacherova, Eva; Bec, Nicole; Anzenbacher, Pavel; Hudecek, Jiri; Soucek, Pavel; Jung, Christiane; Munro, Andrew W.; Lange, ReinhardEuropean Journal of Biochemistry (2000), 267 (10), 2916-2920CODEN: EJBCAI; ISSN:0014-2956. (Blackwell Science Ltd.)The flexibility of the structure and compressibility of the resp. active site of cytochromes P 450 3A4 (CYP3A4) and BM-3 (CYP102) were studied using absorption spectroscopy in the UV and visual regions. Conformational changes in the overall protein structures of both CYP3A4 and CYP102 due to the effects of temp. and pressure are reversible. However, the enzymes differ in the properties of their active sites. The CYP3A4 enzyme denatures to the inactive P420 form relatively easy, at 3000 bar over half is converted to P420. The compressibility of its active site is lower than that of CYP102 and is greater with the substrate bound, which is in line with the obsd. lack of a stabilizing effect of the substrate on its conformation under pressure. In contrast, CYP102, although having the most compressible active site among the P450s, possesses a structure that does not denature easily to the inactive (P420) form under pressure. In this respect, it resembles the P 450 isolated from acidothermophilic archaebacteria.
- 55Harris, H. A.; Albert, L. M.; Leathurby, Y.; Malamas, M. S.; Mewshaw, R. E.; Miller, C. P.; Kharode, Y. P.; Marzolf, J.; Komm, B. S.; Winneker, R. C.; Frail, D. E.; Henderson, R. A.; Zhu, Y.; Keith, J. C. Evaluation of an Estrogen Receptor-β Agonist in Animal Models of Human Disease. Endocrinology 2003, 144, 4241– 4249, DOI: 10.1210/en.2003-0550Google Scholar55https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3sXns1Srt7o%253D&md5=ed1255fd36790a7b9bc665624e5a4619Evaluation of an estrogen receptor-β agonist in animal models of human diseaseHarris, Heather A.; Albert, Leo M.; Leathurby, Yelena; Malamas, Michael S.; Mewshaw, Richard E.; Miller, Chris P.; Kharode, Yogendra P.; Marzolf, James; Komm, Barry S.; Winneker, Richard C.; Frail, Donald E.; Henderson, Ruth A.; Zhu, Yuan; Keith, James C., Jr.Endocrinology (2003), 144 (10), 4241-4249CODEN: ENDOAO; ISSN:0013-7227. (Endocrine Society)The discovery of a second estrogen receptor (ER), called ERβ, in 1996 sparked intense interest within the scientific community to discover its role in mediating estrogen action. However, despite more than 6 yr of research into the function of this receptor, its physiol. role in mediating estrogen action remains unclear and controversial. We have developed a series of highly selective agonists for ERβ and have characterized their activity in several clin. relevant rodent models of human disease. The activity of one such compd., ERB-041, is reported here. We conclude from these studies that ERβ does not mediate the bone-sparing activity of estrogen on the rat skeleton and that it does not affect ovulation or ovariectomy-induced wt. gain. In addn., these compds. are nonuterotrophic and nonmammotrophic. However, ERB-041 has a dramatic beneficial effect in the HLA-B27 transgenic rat model of inflammatory bowel disease and the Lewis rat adjuvant-induced arthritis model. Daily oral doses as low as 1 mg/kg reverse the chronic diarrhea of HLA-B27 transgenic rats and dramatically improve histol. disease scores in the colon. The same dosing regimen in the therapeutic adjuvant-induced arthritis model reduces joint scores from 12 (maximal inflammation) to 1 over a period of 10 d. Synovitis and Mankin (articular cartilage) histol. scores are also significantly lowered (50-75%). These data suggest that one function of ERβ may be to modulate the immune response, and that ERβ-selective ligands may be therapeutically useful agents to treat chronic intestinal and joint inflammation.
- 56Fulmer, G. R.; Miller, A. J. M.; Sherden, N. H.; Gottlieb, H. E.; Nudelman, A.; Stoltz, B. M.; Bercaw, J. E.; Goldberg, K. I. NMR Chemical Shifts of Trace Impurities: Common Laboratory Solvents, Organics, and Gases in Deuterated Solvents Relevant to the Organometallic Chemist. Organometallics 2010, 29, 2176– 2179, DOI: 10.1021/om100106eGoogle Scholar56https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXkslKms7Y%253D&md5=53317e05fe3be77fe7de48d8fe1fb8b7NMR Chemical Shifts of Trace Impurities: Common Laboratory Solvents, Organics, and Gases in Deuterated Solvents Relevant to the Organometallic ChemistFulmer, Gregory R.; Miller, Alexander J. M.; Sherden, Nathaniel H.; Gottlieb, Hugo E.; Nudelman, Abraham; Stoltz, Brian M.; Bercaw, John E.; Goldberg, Karen I.Organometallics (2010), 29 (9), 2176-2179CODEN: ORGND7; ISSN:0276-7333. (American Chemical Society)Tables of 1H and 13C NMR chem. shifts have been compiled for common org. compds. often used as reagents or found as products or contaminants in deuterated org. solvents. Building upon the work of Gottlieb, Kotlyar, and Nudelman in the Journal of Org. Chem., signals for common impurities are now reported in addnl. NMR solvents (tetrahydrofuran-d8, toluene-d8, dichloromethane-d2, chlorobenzene-d5, and 2,2,2-trifluoroethanol-d3) which are frequently used in organometallic labs. Chem. shifts for other orgs. which are often used as reagents or internal stds. or are found as products in organometallic chem. are also reported for all the listed solvents.
- 57Thirumamagal, B. T. S.; Zhao, X. B.; Bandyopadhyaya, A. K.; Johnsamuel, J.; Tiwari, R.; Golightly, D. W.; Patel, V.; Jehning, B. T.; Backer, M. V.; Barth, R. F.; Lee, R. J.; Backer, J. M.; Tjarks, W. Receptor-Targeted Liposomal Delivery of Boron-Containing Cholesterol Mimics for Boron Neutron Capture Therapy (BNCT). Bioconjugate Chem. 2006, 17, 1141– 1150, DOI: 10.1021/bc060075dGoogle Scholar57https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD28XotV2ltbs%253D&md5=4a6a65107118389c48d5ee8233e563f8Receptor-Targeted Liposomal Delivery of Boron-Containing Cholesterol Mimics for Boron Neutron Capture Therapy (BNCT)Thirumamagal, B. T. S.; Zhao, Xiaobin B.; Bandyopadhyaya, Achintya K.; Narayanasamy, Sureshbabu; Johnsamuel, Jayaseharan; Tiwari, Rohit; Golightly, Danold W.; Patel, Vimalkumar; Jehning, Brian T.; Backer, Marina V.; Barth, Rolf F.; Lee, Robert J.; Backer, Joseph M.; Tjarks, WernerBioconjugate Chemistry (2006), 17 (5), 1141-1150CODEN: BCCHES; ISSN:1043-1802. (American Chemical Society)Liposomes have been a main focus of tumor-selective boron delivery strategies in boron neutron capture therapy (BNCT), a binary method for the treatment of cancer that is based on the nuclear reaction between boron atoms and low-energy thermal neutrons. Three novel carboranyl cholesterol derivs. were prepd. as lipid bilayer components for the construction of nontargeted and receptor-targeted boronated liposomes for BNCT. A major structural feature of these novel boronated cholesterol mimics is the replacement of the B and the C ring of cholesterol with a carborane cluster. Computational analyses indicated that all three boronated compds. have structural features and physicochem. properties that are very similar to those of cholesterol. One of the synthesized boronated cholesterol mimics was stably incorporated into non-, folate receptor (FR)-, and vascular endothelial growth factor receptor-2 (VEGFR-2)-targeted liposomes. No major differences were found in appearance, size distribution, and lamellarity between conventional dipalmitoylphosphatidylcholine (DPPC)/cholesterol liposomes, nontargeted, and FR-targeted liposomal formulations of this carboranyl cholesterol deriv. FR-targeted boronated liposomes were taken up extensively in FR overexpressing KB cells in vitro, and the uptake was effectively blocked in the presence of free folate. In contrast, a boronated cholesterol mimic incorporated into nontargeted liposomes showed significantly lower cellular uptake. There was no apparent in vitro cytotoxicity in FR overexpressing KB cells and VEGFR-2 overexpressing 293/KDR cells when these were incubated with boronated FR- and (VEGFR-2)-targeted liposomes, resp., although the former accumulated extensively in KB cells and the latter effectively interacted with VEGFR-2 by causing autophosphorylation and protecting 293/KDR cells from SLT (Shiga-like toxin)-VEGF cytotoxicity.
- 58Fujii, S.; Yamada, A.; Nakano, E.; Takeuchi, Y.; Mori, S.; Masuno, H.; Kagechika, H. Design and Synthesis of Nonsteroidal Progesterone Receptor Antagonists Based on C,C′-Diphenylcarborane Scaffold as a Hydrophobic Pharmacophore. Eur. J. Med. Chem. 2014, 84, 264– 277, DOI: 10.1016/j.ejmech.2014.07.034Google Scholar58https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXht1Grs7bF&md5=5b524301e7d8a908c6848a54652d2dc5Design and synthesis of nonsteroidal progesterone receptor antagonists based on C,C'-diphenylcarborane scaffold as a hydrophobic pharmacophoreFujii, Shinya; Yamada, Ayumi; Nakano, Eiichi; Takeuchi, Yuki; Mori, Shuichi; Masuno, Hiroyuki; Kagechika, HiroyukiEuropean Journal of Medicinal Chemistry (2014), 84 (), 264-277CODEN: EJMCA5; ISSN:0223-5234. (Elsevier Masson SAS)The progesterone receptor (PR) plays important roles in multiple physiol. processes, including female reprodn. Here, we report the synthesis of nonsteroidal PR antagonists contg. a boron cluster as the hydrophobic core. We found that 1,7-diphenyl-meta-carborane was the preferred substructure among the three carborane isomers. One compd. was a potent PR antagonist (IC50: 29 nM). Another compd. also exhibited potent activity (IC50: 93 nM), and did not bind to androgen receptor, glucocorticoid receptor or mineralocorticoid receptor. These compds. may be useful for investigating potential clin. applications of PR modulators.
- 59Sedlak, D.; Paguio, A.; Bartunek, P. Two Panels of Steroid Receptor Luciferase Reporter Cell Lines for Compound Profiling. Comb. Chem. High Throughput Screening 2011, 14, 248– 266, DOI: 10.2174/138620711795222446Google Scholar59https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXnslagtL8%253D&md5=0a021e26cec2615f445f7b3d18625c39Two panels of steroid receptor luciferase reporter cell lines for compound profilingSedlak, David; Paguio, Aileen; Bartunek, PetrCombinatorial Chemistry & High Throughput Screening (2011), 14 (4), 248-266CODEN: CCHSFU; ISSN:1386-2073. (Bentham Science Publishers Ltd.)Steroid hormone receptors represent a major target in drug discovery. As ligand inducible transcription factors, their activity can be modulated by small lipophilic mols. Here we describe two panels of potent and selective luciferase reporter cell lines based on cells with low endogenous steroid receptor activity (U2OS). The panels contain reporter cell lines for estrogen receptors α and β, androgen, glucocorticoid, mineralocorticoid, and progesterone receptors. In the first panel, the activation of either synthetic, steroid response elements contg. promoter or viral promoter is mediated by full-length steroid receptors. The second panel is based on the expression of the chimeric receptor, which was created by the replacement of the N-terminal part of the mol. by Gal4 DBD and that binds to multiple UAS sites in the reporter promoter. Both panels were extensively characterized by profiling 28 ligands in dose response manner in agonist and antagonist mode. We have analyzed and compared the responses to tested ligands from both panels and concluded that in general both systems generated similar qual. response in terms of potency, efficacy, partial agonism/antagonism, mixed agonistic/antagonistic profiles and the rank of potencies was well conserved between both panels. However, we have also identified some artifacts introduced by the Gal4/LBD reporter assays in contrast to their full-length receptor reporter counterparts. Keeping in mind the advantages and drawbacks of each reporter format, these cell lines represent powerful and selective tools for profiling large compd. libraries (HTS) and for detailed study of mechanisms by which compds. exert their biol. effects.
- 60Yung-Chi, C.; Prusoff, W. H. Relationship between the Inhibition Constant (KI) and the Concentration of Inhibitor Which Causes 50 per Cent Inhibition (I50) of an Enzymatic Reaction. Biochem. Pharmacol. 1973, 22, 3099– 3108, DOI: 10.1016/0006-2952(73)90196-2Google ScholarThere is no corresponding record for this reference.
- 61Chang, T. K. H.; Waxman, D. J. Enzymatic Analysis of CDNA-Expressed Human CYP1A1, CYP1A2, and CYP1B1 with 7-Ethoxyresorufin as Substrate. In Cytochrome P450 Protocols; Phillips, I. R.; Shephard, E. A., Eds.; Methods in Molecular Biology; Humana Press: Totowa, NJ, 2006; pp 85– 90.Google ScholarThere is no corresponding record for this reference.
- 62Souček, P. Novel Sensitive High-Performance Liquid Chromatographic Method for Assay of Coumarin 7-Hydroxylation. J. Chromatogr. B: Biomed. Sci. Appl. 1999, 734, 23– 29, DOI: 10.1016/S0378-4347(99)00325-4Google Scholar62https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXns1Ghtbw%253D&md5=313b3ef1e8521fa185c493775b679005Novel sensitive high-performance liquid chromatographic method for assay of coumarin 7-hydroxylationSoucek, PavelJournal of Chromatography B: Biomedical Sciences and Applications (1999), 734 (1), 23-29CODEN: JCBBEP; ISSN:0378-4347. (Elsevier Science B.V.)In this paper, a novel HPLC-based method with fluorometric detection of coumarin 7-hydroxylase is presented. The described method provides a time-effective, more sensitive and specific alternative to the previously used spectrofluorometric assay. Using the developed method, metab. of coumarin in 11 samples of human liver microsomes was evaluated and 1790±690 pmol/min/nmol cytochrome P 450 (CYP) activity was found. Kinetic parameters and linearity of coumarin 7-hydroxylation were studied in a reconstituted system consisting of recombinant CYP2A6 expressed in Escherichia coli, rat NADPH-CYP reductase and usual components. It was found that a 3.5 to 30 min time of incubation is suitable for estn. of coumarin 7-hydroxylase activity. Obsd. Km and Vmax values in the CYP2A6 reconstituted system were 1.48±0.37 μM and 3360±180 pmol product/min/nmol CYP, resp.
- 63Donato, M. T.; Jiménez, N.; Castell, J. V.; Gómez-Lechón, M. J. Fluorescence-Based Assays for Screening Nine Cytochrome P450 (P450) Activities in Intact Cells Expressing Individual Human P450 Enzymes. Drug Metab. Dispos. 2004, 32, 699– 706, DOI: 10.1124/dmd.32.7.699Google Scholar63https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXltlWqt7k%253D&md5=10a4fda9a830c33436a8b2373e3af799Fluorescence-based assays for screening nine cytochrome P450 (P450) activities in intact cells expressing individual human P450 enzymesDonato, M. Teresa; Jimenez, Nuria; Castell, Jose V.; Gomez-Lechon, M. JoseDrug Metabolism and Disposition (2004), 32 (7), 699-706CODEN: DMDSAI; ISSN:0090-9556. (American Society for Pharmacology and Experimental Therapeutics)Here, the authors describe a battery of fluorescence assays for rapid measurement in intact cells of the activity of 9 cytochromes P 450 (P 450) isoforms involved in drug metab. The assays are based on the direct incubation of monolayers of cells expressing individual P 450 enzymes with a fluorogenic substrate followed by fluorometric quantification of the product formed and released into incubation medium. For each individual P 450 activity, different fluorescent probes were examd., and the one showing the best properties (highest metabolic rates, lowest background fluorescence) was selected: 3-cyano-7-ethoxycoumarin for CYP1A2 and CYP2C19, coumarin for CYP2A6, 7-ethoxy-4-trifluoromethylcoumarin for CYP2B6, dibenzylfluorescein for CYP2C8, 7-methoxy-4-trifluoromethylcoumarin (MFC) for CYP2C9 and CYP2E1, 3-[2-(N,N-diethyl-N-methylammonium)ethyl]-7-methoxy-4-methylcoumarin for CYP2D6, and 7-benzyloxy-4-trifluoromethylcoumarin for CYP3A4. The fluorescence-based assays were highly sensitive and allowed the simultaneous measurement of a large no. of samples using plate readers, thus enhancing sample throughput. The major advantages over high-throughput assays in subcellular fractions were that, as living cells were used, manual handling and enzyme damage were minimized, the endoplasmic reticulum of the cells remained intact, exogenous cofactors or NADPH-regenerating systems were not required, and transport processes were maintained. These assays could be applied to preliminary screening of inhibitory effects of new drugs on individual P 450 enzymes. After comparison of the results obtained using the fluorescent probes in intact P 450-expressing cells and those obtained using the HPLC-based selective assays in the same cells, in primary human hepatocytes, or in human liver microsomes, a fairly good agreement was found.
- 64Crespi, C. L.; Chang, T. K. H.; Waxman, D. J. High-Performance Liquid Chromatography Analysis of CYP2C8-Catalyzed Paclitaxel 6α-Hydroxylation. In Cytochrome P450 Protocols; Phillips, I. R.; Shephard, E. A., Eds.; Methods in Molecular Biology; Humana Press: Totowa, NJ, 2006; pp 103– 107.Google ScholarThere is no corresponding record for this reference.
- 65Crespi, C. L.; Chang, T. K. H.; Waxman, D. J. Determination of CYP2C9-Catalyzed Diclofenac 4′-Hydroxylation by High-Performance Liquid Chromatography. In Cytochrome P450 Protocols; Phillips, I. R.; Shephard, E. A., Eds.; Methods in Molecular Biology; Humana Press: Totowa, NJ, 2006; pp 109– 113.Google ScholarThere is no corresponding record for this reference.
- 66Crespi, C. L.; Chang, T. K. H.; Waxman, D. J. CYP2C19-Mediated (S)-Mephenytoin 4′-Hydroxylation Assayed by High-Performance Liquid Chromatography With Radiometric Detection. In Cytochrome P450 Protocols; Phillips, I. R.; Shephard, E. A., Eds.; Methods in Molecular Biology; Humana Press: Totowa, NJ, 2006; pp 115– 119.Google ScholarThere is no corresponding record for this reference.
- 67Crespi, C. L.; Chang, T. K. H.; Waxman, D. J. CYP2D6-Dependent Bufuralol 1′-Hydroxylation Assayed by Reverse-Phase Ion-Pair High-Performance Liquid Chromatography With Fluorescence Detection. In Cytochrome P450 Protocols; Phillips, I. R.; Shephard, E. A., Eds.; Methods in Molecular Biology; Humana Press: Totowa, NJ, 2006; pp 121– 125.Google ScholarThere is no corresponding record for this reference.
- 68Lucas, D.; Menez, J.-F.; Berthou, F. [12] Chlorzoxazone: An in Vitro and in Vivo Substrate Probe for Liver CYP2E1. In Methods in Enzymology; Johnson, E. F.; Waterman, M. R., Eds.; Cytochrome P450, Part B; Academic Press, 1996; Vol. 272, pp 115– 123.Google ScholarThere is no corresponding record for this reference.
- 69Ghosal, A.; Satoh, H.; Thomas, P. E.; Bush, E.; Moore, D. Inhibition and Kinetics of Cytochrome P4503A Activity in Microsomes from Rat, Human, and Cdna-Expressed Human Cytochrome P450. Drug Metab. Dispos. 1996, 24, 940– 947Google Scholar69https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK28XlvV2itrg%253D&md5=fb95979ee683d0166e6159ccd3bee9e2Inhibition and kinetics of cytochrome P4503A activity in microsomes from rat, human, and cDNA-expressed human cytochrome P450Ghosal, Anima; Satoh, Hiroko; Thomas, Paul E.; Bush, Ernest; Moore, DavidDrug Metabolism and Disposition (1996), 24 (9), 940-947CODEN: DMDSAI; ISSN:0090-9556. (Williams & Wilkins)Midazolam (MDZ) is metabolized in human liver microsomes by the cytochrome P 450 (CYP) 3A subfamily to 1'-hydroxy (1'-OH) and 4-hydroxy (4-OH) metabolites. MDZ is metabolized in the rat primarily to 4-OH MDZ, 1'-OH MDZ, and 1',4-dihydroxy (1',4-diOH) MDZ. The kinetics of 4-OH and 1'-OH metabolite formation were detd. using hepatic microsomes from control, Ro 23-7637 and dexamethasone-treated male rats. KM values for the major metabolite, 4-OH MDZ, were 24.5, 43.1, and 32.8 μM, and the corresponding Vmax values were 5.9, 28.9, and 13 nmol/mg/min for the control, DEX, and Ro 23-7637-treated animals, resp. KM values for 1'-hydroxylation of MDZ (the major metabolite) after incubation with human liver microsomes from three individuals were 5.57, 2.50, and 3.56 μM, and the corresponding Vmax values were 4.38, 0.49, and 0.19 nmol/mg/min, resp. In parallel studies using cDNA-expressed human CYP3A4 microsomes, the KM for 1'-OH formation was 1.56 μM, and the corresponding Vmax was 0.16 nmol/mg/min. MDZ was not metabolized by cDNA-expressed human CYP2D6, CYP2E1, or CYP1A2, thus confirming that these isoforms were not responsible for its biotransformation. The formation of 1',4-diOH metabolite in rat and 1'-OH formation in cDNA-expressed human CYP3A4 microsomes showed a decrease in velocity at high substrate concns. Inhibition studies showed that MDZ hydroxylation was strongly inhibited by ketoconazole and Ro 23-7637 in rat, human, and cDNA-expressed human CYP3A4 microsomes. α-Naphthoflavone stimulated 1'-OH metabolite formation in human and cDNA-expressed human CYP3A4 microsomes at low concn. (10 μM). Naringenin, a flavonoid present in grapefruit juice, also inhibited MDZ metab. in human liver microsomes. Immunoinhibition studies revealed that polyclonal anti-rat CYP3A2 antibody inhibited MDZ metab. 80-90% in rat, human, and cDNA-expressed human CYP3A4 microsomes, thus suggesting that members of the CYP3A4 subfamily were involved in the metab.
- 70Kronbach, T.; Mathys, D.; Umeno, M.; Gonzalez, F. J.; Meyer, U. A. Oxidation of Midazolam and Triazolam by Human Liver Cytochrome P450IIIA4. Mol. Pharmacol. 1989, 36, 89– 96Google Scholar70https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaL1MXlt1Kgu7g%253D&md5=6ecf9fc3682977542e46d9a32be16f21Oxidation of midazolam and triazolam by human liver cytochrome P450IIIA4Kronbach, Thomas; Mathys, Daniel; Umeno, Morio; Gonzalez, Frank J.; Meyer, Urs A.Molecular Pharmacology (1989), 36 (1), 89-96CODEN: MOPMA3; ISSN:0026-895X.The metab. of midazolam and triazolam to their 1'-hydroxy and 4-hydroxy metabolites was studied in microsomes of human livers. The formation of both metabolites was inhibited >90% by an antiserum directed against a pregnenolone 16α-carbonitrile-inducible cytochrome P 450 (P450PCN1) of rat liver. Moreover, midazolam hydroxylase activity was immunopptd. from solubilized human microsomes with polyclonal antibodies against rat P450PCN1 and the closely related human isoenzyme P450NF. A close correlation was obsd. between the amt. of protein detected in immunoblots with these antibodies and the midazolam or triazolam or triazolam hydroxylase activity. The formation of both metabolites of midazolam was inhibited by triacetyloleandomycin, a known inhibitor of cytochromes P 450 of the IIIA family. Direct evidence that P450IIIA4 catalyzes the metab. of midazolam was provided through the use of cDNA-directed expression. Monkey COS cells transfected with human P450PCN1 cDNA were able to catalyze both the 1'- and the 4-hydroxylation of midazolam. The metab. of midazolam and triazolam in human liver is apparently predominantly mediated by cytochrome P450IIIA4. Two of 15 human livers expressed a 2nd immunoreactive microsomal protein of higher apparent mol. wt. and were more active in midazolam 1'-hydroxylation. The data also provide evidence that the marked interindividual variation in the response to these widely used benzodiazepine drugs is due to variable hepatic metab.
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Abstract
Figure 1
Figure 1. Estrogen receptor ligands.
Scheme 1
Scheme 1. Synthesis of Target Alkyl- and Hydroxyalkyl-Substituted Carboranesaa(a) n-BuLi, dimethoxyethane (1,2-DME), −78 or 0 °C; (b) BBr3, CH2CI2, 0 °C to room temperature (RT).
Scheme 2
Scheme 2. Synthesis of Ketone 9 and S and R Enantiomers 7 and 8aa(a) PCC, CH2Cl2; (b) BH3·THF, (S)-2-Me-CBS, RT; (c) BH3·THF, (R)-2-Me-CBS, RT; (d) BBr3, CH2Cl2, 0 °C to RT.
Scheme 3
Scheme 3. Synthesis of Phenyl Compound 13aa(a) n-Hexylmagnesium bromide, Et2O, 0 °C; (b) PCC, CH2CI2; (c) BH3·THF, (R)-2-Me-CBS, 0 °C; (d) 1-dodecanethiol, N-methylpyrrolidinone (NMP), NaOH, 100 °C.
Scheme 4
Scheme 4. Synthesis of Cyclohexyl Compound 17aa(a) Cs2CO3, CH3I, acetone, reflux; (b) (methoxymethyl)triphenylphosphonium chloride, LiHMDS, THF; 2 N HCI; (c) hexylmagnesium bromide, Et2O, 0 °C; (d) PCC, CH2CI2; (e) BH3·THF, (R)-2-Me-CBS, 0°C; (f) 1-dodecanethiol, NMP, NaOH, 100 °C.
Scheme 5
Scheme 5. Synthesis of [2.2.2]Bicyclic Compound 26aa(a) Br2, dichloromethane (DCM), HgO; (b) AICI3, benzene, (c) LiAIH4, Et2O; (d) PCC, NaHCO3, NaOAc, CH2CI2; (e) hexylmagnesium bromide, Et2O; (f) AgOAc, Br2, CHCI3; (g) PCC, CH2CI2; (h) BH3·THF, (R)-2-Me-CBS 0 °C; (i) benzaldehyde oxime, Cs2CO3, RockPhos Pd G3.
Scheme 6
Scheme 6. Synthesis of Compounds 28 and 31aa(a) n-BuLi, 1,2-DME, −78 or 0 °C, 1-heptanal; (b) BBr3, CH2CI2, 0 °C to RT.
Figure 2
Figure 2. Ligand selectivity for ERβ increases with elongation of the side hydrocarbon chain. BE120, 3g, and 8 were tested in ERα (dashed line) and ERβ (solid line) transactivation assays in the concentration range between 10 μM and 1 pM, in triplicates and activities were expressed relative to 100 nM 17β-estradiol set to 100%. Increasing length of the carboranes’ side hydrocarbon chain is accompanied with decreasing potency in both ERα and ERβ reporter assays and by increasing selectivity for ERβ (blue: nonselective, green: 10–100× selective, red: selectivity >100×).
Figure 3
Figure 3. Activities of the carboranes’ compound library on steroid receptors. (A) Compounds were profiled at 1 μM in the selective luciferase reporter assays for the agonistic and (B) antagonistic activities for steroid receptors. The activity is expressed relative to maximal activity induced by reference compound or as a fold induction compared to untreated cells for control assay (MMTV reporter alone, dark red dots). Cell viability in U2OS cells was used as a control experiment for the antagonistic activities and is expressed in a scale of 0–100% relative to dimethyl sulfoxide (DMSO)-treated cells (green dots).
References
This article references 70 other publications.
- 1Dodds, E. C.; Lawson, W. Synthetic Strogenic Agents without the Phenanthrene Nucleus. Nature 1936, 137, 996, DOI: 10.1038/137996a01https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaA28XkvVagsQ%253D%253D&md5=9991eb1f0557091ee8f4d26b2c9d99d3Synthetic estrogenic agents without the phenanthrene nucleusDodds, E. C.; Lawson, WilfridNature (London, United Kingdom) (1936), 137 (), 996CODEN: NATUAS; ISSN:0028-0836.The phenanthrene condensed-ring structure is not necessary for estrogenic activity. A table of 14 synthetic substances is given, all except one showing 100% positive estrus responses.
- 2Walter, P.; Green, S.; Greene, G.; Krust, A.; Bornert, J. M.; Jeltsch, J. M.; Staub, A.; Jensen, E.; Scrace, G.; Waterfield, M. Cloning of the Human Estrogen Receptor CDNA. Proc. Natl. Acad. Sci. U.S.A. 1985, 82, 7889– 7893, DOI: 10.1073/pnas.82.23.78892https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaL28XotlShsA%253D%253D&md5=bfb5de85940113236c09e1f34a785b75Cloning of the human estrogen receptor cDNAWalter, Philippe; Green, Stephen; Greene, Geoffrey; Krust, Andree; Bornert, Jean Marc; Jeltsch, Jean Marc; Staub, Adrien; Jensen, Elwood; Scrace, Geoffrey; et al.Proceedings of the National Academy of Sciences of the United States of America (1985), 82 (23), 7889-93CODEN: PNASA6; ISSN:0027-8424.Poly(A)+ RNA isolated from the human breast cancer cell line MCF-7 was fractionated by sucrose gradient centrifugation and fractions enriched in estrogen receptor (ER) mRNA were used to prep. randomly primed cDNA libraries in the λg10 and λg11 vectors. Clones corresponding to ER sequence were isolated from both libraries after screening with either ER monoclonal antibodies (λg11) or synthetic oligonucleotide probes designed from 2 peptide sequences of purified ER (λg10). Five cDNA clones were isolated by antibody screening, and 5 were isolated after screening with synthetic oligonucleotides. The 2 largest ER cDNA clones, λOR3 (1.3 kilobase pairs) and λOR8 (2.1 kilobase pairs), isolated by using antibodies and oligonucleotides, resp., were able to enrich selectively for ER mRNA by hybrid-selection. Further, λOR8 contains the DNA sequence expected from the 2 ER peptides and cross-hybridizes with each of the other ER cDNA clones. These results demonstrate that the clones isolated correspond to the ER mRNA sequence. Use of λOR8 as a hybridization probe revealed a single poly(A)+ RNA band of ≈6.2 kilobase pairs in the ER-contg. human breast cancer cell lines MCF-7 and T47D. In contrast, no hybridization was seen in the human ER-neg. cell line HeLa. The same probe hybridizes to a chicken gene that is expressed in oviduct tissue as a 7.5-kilobase-pair poly(A)+ RNA.
- 3Korach, K. S.; Couse, J. F.; Curtis, S. W.; Washburn, T. F.; Lindzey, J.; Kimbro, K. S.; Eddy, E. M.; Migliaccio, S.; Snedeker, S. M.; Lubahn, D. B.; Schomberg, D. W.; Smith, E. P. Estrogen Receptor Gene Disruption: Molecular Characterization and Experimental and Clinical Phenotypes. Recent Prog. Horm. Res. 1996, 51, 159– 1863https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2sXktFKisr0%253D&md5=b9f0b64fca403a8ac04eff2884ad258aEstrogen receptor gene disruption: molecular characterization and experimental and clinical phenotypesKorach, Kenneth S.; Couse, John F.; Curtis, Sylvia W.; Washburn, Todd F.; Lindzey, Jonathan; Kimbro, K. Sean; Eddy, Edward M.; Migliaccio, Silvia; Snedeker, Suzanne M.; Lubahn, Dennis B.; Schomberg, David W.; Smith, Eric P.Recent Progress in Hormone Research (1996), 51 (), 159-188CODEN: RPHRA6; ISSN:0079-9963. (Endocrine Society)A review with 50 refs. The estrogen receptor (ER) is thought to play a crucial role in the regulation of many life processes, including development, reprodn. and normal physiol. Because there have been no known mutations of the estrogen receptor in normal tissue of humans and animals, its presence and tissue distribution is thought to be essential for survival. Using the techniques of homologous recombination, the ER gene has been disrupted to produce a line of transgenic mice possessing the altered ER gene (ERKO). The mouse ER gene was disrupted by inserting a 1.8-kb PGK-Neomycin sequence into exon 2, ∼280 bp downstream of the transcription start codon. The correct targeting of the disruption was demonstrated by Southern blot anal. and PCR. Western blot anal. of uterine prepns. from ERKO females showed no detectable ER protein. Heterozygotes had one half the level of ER protein compared to wild-type animals. Estrogen insensitivity was confirmed using estrogen agonists, estradiol, hydroxy tamoxifen, diethylstilbestrol treatment for 3 days which resulted in a 3-4-fold increase in uterine wet wt. and vaginal cornification in wild-type females, while ERKO mice were totally unresponsive. These data were further supported by the failure of estrogen or EGF treatment to induce DNA synthesis in uterine tissue of similarly treated mice. Lactoferrin, an estrogen-responsive gene in the uterus, was also assayed by Northern blot. Wild-type mice treated with a single estradiol injection showed a 350-fold induction in lactoferrin mRNA, while ERKO females showed no detectable response. Both male and female animals survive to adulthood with normal gross external phenotypes. As expected, females are infertile and demonstrate hypoplastic uteri and hyperemic ovaries with no apparent corpora lutea. Males are also infertile, with atrophy of the testes and seminiferous tubule dysmorphogenesis. Although the reproductive capabilities have been altered with a dramatic effect on the gonads, prenatal development of the reproductive tracts of both sexes appear to be independent of an ER-mediated response. Anal. of the mammary glands of the ERKO females at 4 mo of age showed a primitive ductal rudiment rather than the fully developed ductal tree seen in wild-type siblings. Also absent were the terminal end buds seen during normal ductal morphogenesis. Both sexes show a decrease in skeletal bone d., supporting a direct role for ER action in bone. A single patient is described who is homozygous for a point mutation in the human ER gene at codon 157. The mutation produces a truncation of the ER protein and results in estrogen insensitivity syndrome. Most significant of the clin. findings are effects on skeletal bone d. and retarded bone age. Findings from the patient and mice suggest that the absence of functional ER is not lethal. Mutation in the ER gene is present in the human population. Further characterization of the mice and identification of addnl. patients will be required to more fully understand the consequences of ER gene mutations.
discussion 186–188.
- 4Kuiper, G. G.; Enmark, E.; Pelto-Huikko, M.; Nilsson, S.; Gustafsson, J. A. Cloning of a Novel Receptor Expressed in Rat Prostate and Ovary. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 5925– 5930, DOI: 10.1073/pnas.93.12.59254https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK28Xjs1Ojs7c%253D&md5=aa1c112de706d680f59afec3e77830b3Cloning of a novel estrogen receptor expressed in rat prostate and ovaryKuiper, George G. J. M.; Enmark, Eva; Pelto-Huikko, Markku; Nilsson, Stefan; Gustafsson, Jan-AakeProceedings of the National Academy of Sciences of the United States of America (1996), 93 (12), 5925-5930CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)The authors have cloned a novel member of the nuclear receptor superfamily. The cDNA of clone 29 was isolated from a rat prostate cDNA library and it encodes a protein of 485 amino acid residues with a calcd. mol. wt. of 54.2 kDa. Clone 29 protein is unique in that it is highly homologous to the rat estrogen receptor (ER) protein, particularly in the DNA-binding domain (95%) and in the C-terminal ligand-binding domain (55%). Expression of clone 29 in rat tissues was investigated by in situ hybridization and prominent expression was found in prostate and ovary. In the prostate clone 29 is expressed in the epithelial cells of the secretory alveoli, whereas in the ovary the granulosa cells in primary, secondary, and mature follicles showed expression of clone 29. Satn. ligand-binding anal. of in vitro synthesized clone 29 protein revealed a single binding component for 17β-estradiol (E2) and high affinity (Kd = 0.6 nM). In ligand-competition expts. the binding affinity decreased in the order E2 > diethylstilbestrol > estriol > estrone > 5α-androstane-3β,17β-diol » testosterone = progesterone = corticosterone = 5α-androstane-3α,17β-diol. In cotransfection expts. of Chinese hamster ovary cells with a clone 29 expression vector and an estrogen-regulated reporter gene, maximal stimulation (about 3-fold) of reporter gene activity was found during incubation with 10 nM of E2. Neither progesterone, testosterone, dexamethasone, thyroid hormone, all-trans-retinoic acid, nor 5α-androstane-3α,17β-diol could stimulate reporter gene activity, whereas estrone and 5α-androstane-3β,17β-diol did. The authors conclude that clone 29 cDNA encodes a novel rat ER, which they suggest be named rat ERβ to distinguish it from the previously cloned ER (ERα) from rat uterus.
- 5Nilsson, S.; Mäkelä, S.; Treuter, E.; Tujague, M.; Thomsen, J.; Andersson, G.; Enmark, E.; Pettersson, K.; Warner, M.; Gustafsson, J.-Å. Mechanisms of Estrogen Action. Physiol. Rev. 2001, 81, 1535– 1565, DOI: 10.1152/physrev.2001.81.4.15355https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD38XislCqtw%253D%253D&md5=e3c660fbdf99d1b456486caabef516c8Mechanisms of estrogen actionNilsson, Stefan; Makela, Sari; Treuter, Eckardt; Tujague, Michel; Thomsen, Jane; Andersson, Goran; Enmark, Eva; Pettersson, Katarina; Warner, Margaret; Gustafsson, Jan-AkePhysiological Reviews (2001), 81 (4), 1535-1565CODEN: PHREA7; ISSN:0031-9333. (American Physiological Society)A review. Our appreciation of the physiol. functions of estrogens and the mechanisms through which estrogens bring about these functions has changed during the past decade. Just as transgenic mice were produced in which estrogen receptors had been inactivated and we thought that we were about to understand the role of estrogen receptors in physiol. and pathol., it was found that there was not 1 but 2 distinct and functional estrogen receptors, now called ERα and ERβ. Transgenic mice in which each of the receptors or both the receptors are inactive have revealed a much broader role for estrogens in the body than was previously thought. This decade also saw the description of a male patient who had no functional ERα and whose continued bone growth clearly revealed an important function of estrogen in men. The importance of estrogen in both males and females was also demonstrated in the lab. in transgenic mice in which the aromatase gene was inactivated. Finally, crystal structures of the estrogen receptors with agonists and antagonists have revealed much about how ligand binding influences receptor conformation and how this conformation influences interaction of the receptor with coactivators or corepressors and hence dets. cellular response to ligands.
- 6Heldring, N.; Pike, A.; Andersson, S.; Matthews, J.; Cheng, G.; Hartman, J.; Tujague, M.; Ström, A.; Treuter, E.; Warner, M.; Gustafsson, J.-Å. Estrogen Receptors: How Do They Signal and What Are Their Targets. Physiol. Rev. 2007, 87, 905– 931, DOI: 10.1152/physrev.00026.20066https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXptFGls7k%253D&md5=207d16fa37d6b14483b9fe1002ce2646Estrogen receptors: how do they signal and what are their targetsHeldring, Nina; Pike, Ashley; Andersson, Sandra; Matthews, Jason; Cheng, Guojun; Hartman, Johan; Tujague, Michel; Stroem, Anders; Treuter, Eckardt; Warner, Margaret; Gustafsson, Jan-AakePhysiological Reviews (2007), 87 (3), 905-931CODEN: PHREA7; ISSN:0031-9333. (American Physiological Society)A review. During the past decade there has been a substantial advance in our understanding of estrogen signaling both from a clin. as well as a preclin. perspective. Estrogen signaling is a balance between two opposing forces in the form of two distinct receptors (ERα and ERβ) and their splice variants. The prospect that these two pathways can be selectively stimulated or inhibited with subtype-selective drugs constitutes new and promising therapeutic opportunities in clin. areas as diverse as hormone replacement, autoimmune diseases, prostate and breast cancer, and depression. Mol. biol., biochem., and structural studies have generated information which is invaluable for the development of more selective and effective ER ligands. We have also become aware that ERs do not function by themselves but require a no. of coregulatory proteins whose cell-specific expression explains some of the distinct cellular actions of estrogen. Estrogen is an important morphogen, and many of its proliferative effects on the epithelial compartment of glands are mediated by growth factors secreted from the stromal compartment. Thus understanding the cross-talk between growth factor and estrogen signaling is essential for understanding both normal and malignant growth. In this review we focus on several of the interesting recent discoveries concerning estrogen receptors, on estrogen as a morphogen, and on the mol. mechanisms of anti-estrogen signaling.
- 7Carmeci, C.; Thompson, D. A.; Ring, H. Z.; Francke, U.; Weigel, R. J. Identification of a Gene (GPR30) with Homology to the G-Protein-Coupled Receptor Superfamily Associated with Estrogen Receptor Expression in Breast Cancer. Genomics 1997, 45, 607– 617, DOI: 10.1006/geno.1997.49727https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2sXnsVejsbo%253D&md5=831bb2d172786fe61c914e8e851d0735Identification of a gene (GPR30) with homology to the G-protein-coupled receptor superfamily associated with estrogen receptor expression in breast cancerCarmeci, Charles; Thompson, Devon A.; Ring, Huijun Z.; Francke, Uta; Weigel, Ronald J.Genomics (1997), 45 (3), 607-617CODEN: GNMCEP; ISSN:0888-7543. (Academic Press)Using the technique of differential cDNA library screening, a cDNA clone was isolated from an estrogen receptor (ER)-pos. breast carcinoma cell line (MCF7) cDNA library based upon the overexpression of this gene compared to an ER-neg. cell line (MDA-MB-231). Sequence anal. of this clone detd. that it shared significant homol. to G-protein-coupled receptors. This receptor, GPCR-Br, was abundantly expressed in the ER-pos. breast carcinoma cell lines MCF7, T-47D, and MDA-MB-361. Expression was absent or minimal in the ER-neg. breast carcinoma cell lines BT-20, MDA-MB-231, and HBL-100. GPCR-Br was ubiquitously expressed in human tissues examd. but was most abundant in placenta. GPCR-Br expression was examd. in 11 primary breast carcinomas. GPCR-Br was detected in all 4 ER-pos. tumors and only 1 of 7 ER-neg. tumors. Based upon PCR anal. in hybrid cell lines, the gene for GPCR-Br (HGMW-approved symbol GPR30) was mapped to chromosome 7p22. The pattern of expression of GPCR-Br indicates that this receptor may be involved in physiol. responses specific to hormonally responsive tissues.
- 8Revankar, C. M.; Cimino, D. F.; Sklar, L. A.; Arterburn, J. B.; Prossnitz, E. R. A Transmembrane Intracellular Estrogen Receptor Mediates Rapid Cell Signaling. Science 2005, 307, 1625– 1630, DOI: 10.1126/science.11069438https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXitV2is7c%253D&md5=938d5a0422146eac0ace28da892d82efA Transmembrane Intracellular Estrogen Receptor Mediates Rapid Cell SignalingRevankar, Chetana M.; Cimino, Daniel F.; Sklar, Larry A.; Arterburn, Jeffrey B.; Prossnitz, Eric R.Science (Washington, DC, United States) (2005), 307 (5715), 1625-1630CODEN: SCIEAS; ISSN:0036-8075. (American Association for the Advancement of Science)The steroid hormone estrogen regulates many functionally unrelated processes in numerous tissues. Although it is traditionally thought to control transcriptional activation through the classical nuclear estrogen receptors, it also initiates many rapid nongenomic signaling events. We found that of all G protein-coupled receptors characterized to date, GPR30 is uniquely localized to the endoplasmic reticulum, where it specifically binds estrogen and fluorescent estrogen derivs. Activating GPR30 by estrogen resulted in intracellular calcium mobilization and synthesis of phosphatidylinositol 3,4,5-trisphosphate in the nucleus. Thus, GPR30 represents an intracellular transmembrane estrogen receptor that may contribute to normal estrogen physiol. as well as pathophysiol.
- 9Kuiper, G. G. J. M.; Carlsson, B.; Grandien, K.; Enmark, E.; Häggblad, J.; Nilsson, S.; Gustafsson, J.-Å. Comparison of the Ligand Binding Specificity and Transcript Tissue Distribution of Estrogen Receptors α and β. Endocrinology 1997, 138, 863– 870, DOI: 10.1210/endo.138.3.49799https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2sXht1OksrY%253D&md5=bb4d88e7ba9c878a79c711b3b0a5f92fComparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors α and βKuiper, George G. J. M.; Carlsson, Bo; Grandien, Kaj; Enmark, Eva; Haeggblad, Johan; Nilsson, Stefan; Gustafsson, Jan-AakeEndocrinology (1997), 138 (3), 863-870CODEN: ENDOAO; ISSN:0013-7227. (Endocrine Society)The rat estrogen receptor (ER) exists as two subtypes, ERα and ERβ, which differ in the C-terminal ligand-binding domain and in the N-terminal transactivation domain. In this study we investigated the mRNA expression of both ER subtypes in rat tissues by RT-PCR and compared the ligand binding specificity of the ER subtypes. Satn. ligand binding anal. of in vitro synthesized human ERα and rat ERβ protein revealed a single binding component for 16α-iodo-17β-estradiol with high affinity [dissocn. const. (Kd) = 0.1 nM for ERα protein and 0.4 nM for ERβ protein]. Most estrogenic substrates or estrogenic antagonists compete with 16α-[125I]iodo-17β-estradiol for binding to both ER subtypes in a very similar preference and degree; i.e., diethylstilbestrol > hexestrol > dienestrol > 4-OH-tamoxifen > 17β-estradiol > coumestrol, ICI-164384 > estrone, 17α-estradiol > nafoxidine, moxestrol > clomifene > estriol, 4-OH-estradiol > tamoxifen, 2-OH-estradiol, 5-androstene-3β-17β-diol, genistein for the ERα protein and dienestrol > 4-OH-tamoxifen > diethylstilbestrol > hexestrol > coumestrol, ICI-164384 > 17β-estradiol > estrone, genistein > estriol > nafoxidine, 5-androstene-3β,17β-diol > 17α-estradiol, clomifene, 2-OH-estradiol > 4-OH-estradiol, tamoxifen, moxestrol for the ERβ protein. The rat tissue distribution and/or the relative level of ERα and ERβ expression seems to be quite different, i.e. moderate to high expression in uterus, testis, pituitary, ovary, kidney, epididymis, and adrenal for ERα and prostate, ovary, lung, bladder, brain, uterus, and testis for ERβ. The described differences between the ER subtypes in relative ligand binding affinity and tissue distribution could contribute to the selective action of ER agonists and antagonists in different tissues.
- 10Hamilton, K. J.; Hewitt, S. C.; Arao, Y.; Korach, K. S. Estrogen Hormone Biology. In Current Topics in Developmental Biology; Forrest, D.; Tsai, S., Eds.; Nuclear Receptors in Development and Disease; Academic Press: 2017; Chapter 4; Vol. 125, pp 109– 146.There is no corresponding record for this reference.
- 11Zimmerman, M. A.; Budish, R. A.; Kashyap, S.; Lindsey, S. H. GPER–Novel Membrane Oestrogen Receptor. Clin. Sci. 2016, 130, 1005– 1016, DOI: 10.1042/CS2016011411https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XotVSrsLs%253D&md5=7404f65583d895b9ccc19505eb9a858bGPER-novel membrane estrogen receptorZimmerman, Margaret A.; Budish, Rebecca A.; Kashyap, Shreya; Lindsey, Sarah H.Clinical Science (2016), 130 (12), 1005-1016CODEN: CSCIAE; ISSN:1470-8736. (Portland Press Ltd.)The recent discovery of the G protein-coupled estrogen receptor (GPER) presents new challenges and opportunities for understanding the physiol., pathophysiol. and pharmacol. of many diseases. This review will focus on the expression and function of GPER in hypertension, kidney disease, atherosclerosis, vascular remodelling, heart failure, reprodn., metabolic disorders, cancer, environmental health and menopause. Furthermore, this review will highlight the potential of GPER as a therapeutic target.
- 12Mäkelä, S.; Savolainen, H.; Aavik, E.; Myllärniemi, M.; Strauss, L.; Taskinen, E.; Gustafsson, J.-Å.; Häyry, P. Differentiation between Vasculoprotective and Uterotrophic Effects of Ligands with Different Binding Affinities to Estrogen Receptors α and β. Proc. Natl. Acad. Sci. U.S.A. 1999, 96, 7077– 7082, DOI: 10.1073/pnas.96.12.707712https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXks1WktbY%253D&md5=eac9076dd6e164de1216e54059d9782cDifferentiation between vasculoprotective and uterotrophic effects of ligands with different binding affinities to estrogen receptors α and βMakela, Sari; Savolainen, Hanna; Aavik, Einari; Myllarniemi, Marjukka; Strauss, Leena; Taskinen, Eero; Gustafsson, Jan-Ake; Hayry, PekkaProceedings of the National Academy of Sciences of the United States of America (1999), 96 (12), 7077-7082CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)Estrogen-based drug therapy in cardiovascular diseases has been difficult because it has not been possible to sep. the wanted vasculoprotective effect from the unwanted effects of the hormone to the reproductive system. Here we demonstrate that, after endothelial denudation of rat carotid artery, the mRNA of the classical estrogen receptor (ER) is constitutively expressed at a low level, whereas the expression of the novel ER mRNA increases >40-fold. Under in situ hybridization and immunohistochem., ERβ mRNA and protein colocalize with the smooth muscle cells in the media and neointima. Treatment of ovariectomized female rats with the isoflavone phytoestrogen genistein, which shows 20-fold higher binding affinity to ERβ than to ERα, or with 17β-estradiol, which does not differentiate between the two receptors, provides similar dose-dependent vasculoprotective effect in rat carotid injury model. In addn. in concn. <10 μM, both ligands are equally inhibitory to the replication and migration of smooth muscle cells in vitro. However, only treatment with 17β-estradiol, but not with genistein, is accompanied with a dose-dependent uterotrophic effect. The results suggest that preferential targeting to ERβ will provide vasculoprotective estrogen analogs devoid of effects to the reproductive system.
- 13Thomas, P.; Pang, Y.; Filardo, E. J.; Dong, J. Identity of an Estrogen Membrane Receptor Coupled to a G Protein in Human Breast Cancer Cells. Endocrinology 2005, 146, 624– 632, DOI: 10.1210/en.2004-106413https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXotlWitw%253D%253D&md5=6870880ddbd84da144dc18d929707bcdIdentity of an estrogen membrane receptor coupled to a G protein in human breast cancer cellsThomas, P.; Pang, Y.; Filardo, E. J.; Dong, J.Endocrinology (2005), 146 (2), 624-632CODEN: ENDOAO; ISSN:0013-7227. (Endocrine Society)Although nonclassical estrogen actions initiated at the cell surface have been described in many tissues, the identities of the membrane estrogen receptors (mERs) mediating these actions remain unclear. Here we show that GPR30, an orphan receptor unrelated to nuclear estrogen receptors, has all the binding and signaling characteristics of a mER. A high-affinity (dissocn. const. 2.7 nM), limited capacity, displaceable, single binding site specific for estrogens was detected in plasma membranes of SKBR3 breast cancer cells that express GPR30 but lack nuclear estrogen receptors. Progesterone-induced increases and small interfering RNA-induced decreases in GPR30 expression in SKBR3 cells were accompanied by parallel changes in specific estradiol-17β (E2) binding. Plasma membranes of human embryonic kidney 293 cells transfected with GPR30, but not those of untransfected cells, and human placental tissues that express GPR30 also displayed high-affinity, specific estrogen binding typical of mERs. E2 treatment of transfected cell membranes caused activation of a stimulatory G protein that is directly coupled to the receptor, indicating GPR30 is a G protein-coupled receptor (GPCR), and also increased adenylyl cyclase activity. The finding that the antiestrogens tamoxifen and ICI 182,780, and an environmental estrogen, ortho, para-dichlorodiphenyl-dichloroethylene (o,p'-DDE), have high binding affinities to the receptor and mimic the actions of E2 has important implications for both the development and treatment of estrogen-dependent breast cancer. GPR30 is structurally unrelated to the recently discovered family of GPCR-like membrane progestin receptors. The identification of a second distinct class of GPCR-like steroid membrane receptors suggests a widespread role for GPCRs in nonclassical steroid hormone actions.
- 14Lubahn, D. B.; Moyer, J. S.; Golding, T. S.; Couse, J. F.; Korach, K. S.; Smithies, O. Alteration of Reproductive Function but Not Prenatal Sexual Development after Insertional Disruption of the Mouse Estrogen Receptor Gene. Proc. Natl. Acad. Sci. U.S.A. 1993, 90, 11162– 11166, DOI: 10.1073/pnas.90.23.1116214https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2cXks1ajsw%253D%253D&md5=bde5d2b52c3e2fda7c834175b3e61ca0Alteration of reproductive function but not prenatal sexual development after insertional disruption of the mouse estrogen receptor geneLubahn, Dennis B.; Moyer, Jeffrey S.; Golding, Thomas S.; Couse, John F.; Korach, Kenneth S.; Smithies, OliverProceedings of the National Academy of Sciences of the United States of America (1993), 90 (23), 11162-6CODEN: PNASA6; ISSN:0027-8424.Estrogen receptor and its ligand, estradiol, have long been thought to be essential for survival, fertility, and female sexual differentiation and development. Consistent with this proposed crucial role, no human estrogen receptor gene mutations are known, unlike the androgen receptor, where many loss of function mutations have been found. The authors have generated mutant mice lacking responsiveness to estradiol by disrupting the estrogen receptor gene by gene targeting. Both male and female animals survive to adulthood with normal gross external phenotypes. Females are infertile; males have a decreased fertility. Females have hypoplastic uteri and hyperemic ovaries with no detectable corpora lutea. In adult wild-type and heterozygous females, 3-day estradiol treatment at 40 μg/kg stimulates a 3- to 4-fold increase in uterine wet wt. and alters vaginal cornification, but the uteri and vagina do not respond in the animals with the estrogen receptor gene disruption. Prenatal male and female reproductive tract development can therefore occur in the absence of estradiol receptor-mediated responsiveness.
- 15Krege, J. H.; Hodgin, J. B.; Couse, J. F.; Enmark, E.; Warner, M.; Mahler, J. F.; Sar, M.; Korach, K. S.; Gustafsson, J.-Å.; Smithies, O. Generation and Reproductive Phenotypes of Mice Lacking Estrogen Receptor β. Proc. Natl. Acad. Sci. U.S.A. 1998, 95, 15677– 15682, DOI: 10.1073/pnas.95.26.1567715https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXhvFeiuw%253D%253D&md5=50cc5538ab791d52f280959e296d0e8dGeneration and reproductive phenotypes of mice lacking estrogen receptor βKrege, John H.; Hodgin, Jeffrey B.; Couse, John F.; Enmark, Eva; Warner, Margaret; Mahler, Joel F.; Sar, Madhabananda; Korach, Kenneth S.; Gustafsson, Jan-Ake; Smithies, OliverProceedings of the National Academy of Sciences of the United States of America (1998), 95 (26), 15677-15682CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)Estrogens influence the differentiation and maintenance of reproductive tissues and affect lipid metab. and bone remodeling. Two estrogen receptors (ERs) have been identified to date, ERα and ERβ. The authors previously generated and studied knockout mice lacking estrogen receptor α and reported severe reproductive and behavioral phenotypes including complete infertility of both male and female mice and absence of breast tissue development. Here, the authors describe the generation of mice lacking estrogen receptor β (ERβ -/-) by insertion of a neomycin-resistance gene into exon 3 of the coding gene by using homologous recombination in embryonic stem cells. Mice lacking this receptor develop normally and are indistinguishable grossly and histol. as young adults from their littermates. RNA anal. and immunocytochem. show that tissues from ERβ -/- mice lack normal ERβ RNA and protein. Breeding expts. with young, sexually mature females show that they are fertile and exhibit normal sexual behavior, but have fewer and smaller litters than wild-type mice. Superovulation expts. indicate that this redn. in fertility is the result of reduced ovarian efficiency. The mutant females have normal breast development and lactate normally. Young, sexually mature male mice show no overt abnormalities and reproduce normally. Older mutant males display signs of prostate and bladder hyperplasia. Thus, ERβ is essential for normal ovulation efficiency but is not essential for female or male sexual differentiation, fertility, or lactation. Future expts. are required to det. the role of ERβ in bone and cardiovascular homeostasis.
- 16Prossnitz, E. R.; Hathaway, H. J. What Have We Learned about GPER Function in Physiology and Disease from Knockout Mice?. J. Steroid Biochem. Mol. Biol. 2015, 153, 114– 126, DOI: 10.1016/j.jsbmb.2015.06.01416https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2MXht1CkurnN&md5=76ffa0a4124d62506b9add2f0459dd32What have we learned about GPER function in physiology and disease from knockout mice?Prossnitz, Eric R.; Hathaway, Helen J.Journal of Steroid Biochemistry and Molecular Biology (2015), 153 (), 114-126CODEN: JSBBEZ; ISSN:0960-0760. (Elsevier Ltd.)Estrogens, predominantly 17β-estradiol, exert diverse effects throughout the body in both normal and pathophysiol., during development and in reproductive, metabolic, endocrine, cardiovascular, nervous, musculoskeletal and immune systems. Estrogen and its receptors also play important roles in carcinogenesis and therapy, particularly for breast cancer. In addn. to the classical nuclear estrogen receptors (ERα and ERβ) that traditionally mediate predominantly genomic signaling, the G protein-coupled estrogen receptor GPER has become recognized as a crit. mediator of rapid signaling in response to estrogen. Mouse models, and in particular knockout (KO) mice, represent an important approach to understand the functions of receptors in normal physiol. and disease. Whereas ERα KO mice display multiple significant defects in reprodn. and mammary gland development, ERβ KO phenotypes are more limited, and GPER KO exhibit no reproductive deficits. However, the study of GPER KO mice over the last six years has revealed that GPER deficiency results in multiple physiol. alterations including obesity, cardiovascular dysfunction, insulin resistance and glucose intolerance. In addn., the lack of estrogen-mediated effects in numerous tissues of GPER KO mice, studied in vivo or ex vivo, including those of the cardiovascular, endocrine, nervous and immune systems, reveals GPER as a genuine mediator of estrogen action. Importantly, GPER KO mice have also demonstrated roles for GPER in breast carcinogenesis and metastasis. In combination with the supporting effects of GPER-selective ligands and GPER knockdown approaches, GPER KO mice demonstrate the therapeutic potential of targeting GPER activity in diseases as diverse as obesity, diabetes, multiple sclerosis, hypertension, atherosclerosis, myocardial infarction, stroke and cancer.
- 17Harris, H. A. Estrogen Receptor-β: Recent Lessons from in Vivo Studies. Mol. Endocrinol. 2007, 21, 1– 13, DOI: 10.1210/me.2005-045917https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXmsFOnsw%253D%253D&md5=f55e31161a74e96fa82518f7aac9e62fEstrogen receptor-β: recent lessons from in vivo studiesHarris, Heather A.Molecular Endocrinology (2007), 21 (1), 1-13CODEN: MOENEN; ISSN:0888-8809. (Endocrine Society)A review. The unexpected discovery of a second form of the estrogen receptor (ER), designated ERβ, surprised and energized the field of estrogen research. In the 9 yr since its identification, the remarkable efforts from academic and industrial scientists of many disciplines have made significant progress in elucidating its biol. A powerful battery of tools, including knockout mice as well as a panel of receptor-selective agonists, has allowed an investigation into the role of ERβ. To date, in vivo efficacy studies are limited to rodents. Current data indicate that ERβ plays a minor role in mediating estrogen action in the uterus, on the hypothalamus/pituitary, the skeleton, and other classic estrogen target tissues. However, a clear role for ERβ has been established in the ovary, cardiovascular system, and brain as well as in several animal models of inflammation including arthritis, endometriosis, inflammatory bowel disease, and sepsis. The next phase of research will focus on elucidating, at a mol. level, how ERβ exerts these diverse effects and exploring the clin. utility of ERβ-selective agonists.
- 18Warner, M.; Wu, W.; Montanholi, L.; Nalvarte, I.; Antonson, P.; Gustafsson, J.-A. Ventral Prostate and Mammary Gland Phenotype in Mice with Complete Deletion of the ERβ Gene. Proc. Natl. Acad. Sci. U.S.A. 2020, 117, 4902– 4909, DOI: 10.1073/pnas.192047811718https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXksVSrur0%253D&md5=5b9cf899ca7c8779041f68a6164349d1Ventral prostate and mammary gland phenotype in mice with complete deletion of the ERβ geneWarner, Margaret; Wu, Wan-fu; Montanholi, Leticia; Nalvarte, Ivan; Antonson, Per; Gustafsson, Jan-AkeProceedings of the National Academy of Sciences of the United States of America (2020), 117 (9), 4902-4909CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)Disagreements about the phenotype of estrogen receptor β; (ERβ) knockout mouse, created by removing the DNA-binding domain of the ERβ; gene or interruption of the gene with a neocassette (Oliver Smithies ERβ knockout mice [ERβOS-/-]), prompted us to create an ERβ knockout mouse by deleting the ERβ gene with the use of CRISPR/Cas9 technol. We confirmed that the ERβ gene was eliminated from the mouse genome and that no ERβ mRNA or protein was detectable in tissues of this mouse. Overall the phenotype of the ventral prostate (VP) and mammary gland (MG) in ERβcrispr-/- mice was similar to, but more severe than, that in the ERβOS-/-mice. In the VP of 6-mo-old ERβcrispr-/- mice there was epithelial hyperplasia, fibroplasia, inflammation, stromal overgrowth, and intraductal cancer-like lesions. This was accompanied by an increase in Ki67 and P63 and loss in DACH1 and PURα, two androgen receptor (AR) repressors. In the MG there was overexpression of estrogen receptor α and progesterone receptor, loss of collagen, increase in proliferation and expression of metalloproteases, and invasive epithelium. Surprisingly, by 18 mo of age, the no. of hyperplastic foci was reduced, the ducts of the VP and MG became atrophic, and, in the VP, there was massive immune infiltration and massive desquamation of the luminal epithelial cells. These changes were coincident with reduced levels of androgens in males and estrogens in females. We conclude that ERβ is a tumor suppressor gene in the VP and MG where its loss increases the activity AR and ERα resp.
- 19Meyers, M. J.; Sun, J.; Carlson, K. E.; Marriner, G. A.; Katzenellenbogen, B. S.; Katzenellenbogen, J. A. Estrogen Receptor-β Potency-Selective Ligands: Structure–Activity Relationship Studies of Diarylpropionitriles and Their Acetylene and Polar Analogues. J. Med. Chem. 2001, 44, 4230– 4251, DOI: 10.1021/jm010254a19https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3MXnsFagsr8%253D&md5=8fbf46c36aaf2ffcc9a8c44b15827bfeEstrogen Receptor-β Potency-Selective Ligands: Structure-Activity Relationship Studies of Diarylpropionitriles and Their Acetylene and Polar AnaloguesMeyers, Marvin J.; Sun, Jun; Carlson, Kathryn E.; Marriner, Gwendolyn A.; Katzenellenbogen, Benita S.; Katzenellenbogen, John A.Journal of Medicinal Chemistry (2001), 44 (24), 4230-4251CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)Through an effort to develop novel ligands that have subtype selectivity for the estrogen receptors alpha (ERα) and beta (ERβ), we have found that 2,3-bis(4-hydroxyphenyl)propionitrile (DPN) acts as an agonist on both ER subtypes, but has a 70-fold higher relative binding affinity and 170-fold higher relative potency in transcription assays with ERβ than with ERα. To investigate the ERβ affinity- and potency-selective character of this DPN further, we prepd. a series of DPN analogs in which both the ligand core and the arom. rings were modified by the repositioning of phenolic hydroxy groups and by the addn. of alkyl substituents and nitrile groups. We also prepd. other series of DPN analogs in which the nitrile functionality was replaced with acetylene groups or polar functions, to mimic the linear geometry or polarity of the nitrile, resp. To varying degrees, all of the analogs show preferential binding affinity for ERβ (i.e., they are ERβ affinity-selective), and many, but not all of them, are also more potent in activating transcription through ERβ than through ERα (i.e., they are ERβ potency-selective). meso-2,3-Bis(4-hydroxyphenyl)succinonitrile and dl-2,3-bis(4-hydroxyphenyl)succinonitrile are among the highest ERβ affinity-selective ligands, and they have an ERβ potency selectivity that is equiv. to that of DPN. The acetylene analogs have higher binding affinities but somewhat lower selectivities than their nitrile counterparts. The polar analogs have lower affinities, and only the fluorinated polar analogs have substantial affinity selectivities. This study suggests that, in this series of ligands, the nitrile functionality is crit. to ERβ selectivity because it provides the optimal combination of linear geometry and polarity. Furthermore, the addn. of a second nitrile group β to the nitrile in DPN or the addn. of a Me substituent at an ortho position on the β-arom. ring increases the affinity and selectivity of these compds. for ERβ. These ERβ-selective compds. may prove to be valuable tools in understanding the differences in structure and biol. function of ERα and ERβ.
- 20Stauffer, S. R.; Coletta, C. J.; Tedesco, R.; Nishiguchi, G.; Carlson, K.; Sun, J.; Katzenellenbogen, B. S.; Katzenellenbogen, J. A. Pyrazole Ligands: Structure–Affinity/Activity Relationships and Estrogen Receptor-α-Selective Agonists. J. Med. Chem. 2000, 43, 4934– 4947, DOI: 10.1021/jm000170m20https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3cXot1yjsbk%253D&md5=3bce249ad2d0eaf91aa4ad17ebffdd05Pyrazole Ligands: Structure-Affinity/Activity Relationships and Estrogen Receptor-α-Selective AgonistsStauffer, Shaun R.; Coletta, Christopher J.; Tedesco, Rosanna; Nishiguchi, Gisele; Carlson, Kathryn; Sun, Jun; Katzenellenbogen, Benita S.; Katzenellenbogen, John A.Journal of Medicinal Chemistry (2000), 43 (26), 4934-4947CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)We have found that certain tetrasubstituted pyrazoles are high-affinity ligands for the estrogen receptor (ER) and that one pyrazole is considerably more potent as an agonist on the ERα than on the ERβ subtype. To investigate what substituent pattern provides optimal ER binding affinity and the greatest enhancement of potency as an ERα-selective agonist, we prepd. a no. of tetrasubstituted pyrazole analogs with defined variations at certain substituent positions. Anal. of their binding affinity pattern shows that a C(4)-Pr substituent is optimal and that a p-hydroxyl group on the N(1)-Ph group also enhances affinity and selectivity for ERα. The best compd. in this series, a propylpyrazole triol (I), binds to ERα with high affinity (ca. 50% that of estradiol), and it has a 410-fold binding affinity preference for ERα. It also activates gene transcription only through ERα. Thus, this compd. represents the first ERα-specific agonist. We investigated the mol. basis for the exceptional ERα binding affinity and potency selectivity of pyrazole I by a further study of structure-affinity relationships in this series and by mol. modeling. These investigations suggest that the pyrazole triols prefer to bind to ERα with their C(3)-phenol in the estradiol A-ring binding pocket and that binding selectivity results from differences in the interaction of the pyrazole core and C(4)-Pr group with portions of the receptor where ERα has a smaller residue than ERβ. These ER subtype-specific interactions and the ER subtype-selective ligands that can be derived from them should prove useful in defining those biol. activities in estrogen target cells that can be selectively activated through ERα.
- 21Warner, M.; Huang, B.; Gustafsson, J.-A. Estrogen Receptor β as a Pharmaceutical Target. Trends Pharmacol. Sci. 2017, 38, 92– 99, DOI: 10.1016/j.tips.2016.10.00621https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XhslKnu7jM&md5=976f8a25342bba055e9ba332c84f7ff2Estrogen Receptor β as a Pharmaceutical TargetWarner, Margaret; Huang, Bo; Gustafsson, Jan-AkeTrends in Pharmacological Sciences (2017), 38 (1), 92-99CODEN: TPHSDY; ISSN:0165-6147. (Elsevier Ltd.)A major issue in clin. endocrinol. today is how to use hormones to achieve the health benefits that they clearly can provide but avoid the neg. side effects, i.e., how to develop more precise medicines. This problem of how to use hormones is pervasive in clin. endocrinol. It is true for estrogen, progesterone, androgen, vitamin D, and thyroid hormone, and the problem is amplified in the case of new ligands for the more recently discovered nuclear receptors. Selective targeting of hormone receptor subtypes is one attractive way to harness the beneficial effects of hormones while reducing unwanted side effects. Here, we focus on estrogen receptor (ER)β, which has promise as a selective target in hormone replacement therapy, and in breast and prostate cancers.
- 22Enmark, E.; Gustafsson, J.-Å. Oestrogen Receptors – an Overview. J. Intern. Med. 1999, 246, 133– 138, DOI: 10.1046/j.1365-2796.1999.00545.x22https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXmtlCmtb8%253D&md5=731a0a091639714ccb2a4b8760eec5a6Oestrogen receptors - an overviewEnmark, E.; Gustafsson, J.-A.Journal of Internal Medicine (1999), 246 (2), 133-138CODEN: JINMEO; ISSN:0954-6820. (Blackwell Science Ltd.)A review with 29 refs. The estrogen receptor (ER) is a ligand-activated transcription factor that mediates the effects of the steroid hormone 17β-estradiol in both males and females. Since the isolation and cloning of ER, the prevailing opinion has been that only one such receptor exists. The finding of a second subtype of ER (ERβ) has caused considerable excitement and has forced endocrinologists to re-evaluate many aspects of the actions of estrogens. In this article, the authors will try to summarize the current knowledge about the two estrogen receptor subtypes, with the emphasis on estrogen receptor β (ERβ), and to comment on the observations in mice lacking either receptor or the hormone itself.
- 23Harris, H. A.; Bapat, A. R.; Gonder, D. S.; Frail, D. E. The Ligand Binding Profiles of Estrogen Receptors α and β Are Species Dependent. Steroids 2002, 67, 379– 384, DOI: 10.1016/S0039-128X(01)00194-523https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD38XhslWgs7g%253D&md5=7d8abb8e1b0083d859b19556dcfa4355The ligand binding profiles of estrogen receptors α and β are species dependentHarris, Heather A.; Bapat, Ashok R.; Gonder, Daniel S.; Frail, Donald E.Steroids (2002), 67 (5), 379-384CODEN: STEDAM; ISSN:0039-128X. (Elsevier Science Inc.)Estrogens and selective estrogen receptor modulators are used for the treatment and prevention of conditions resulting from menopause. Since estrogens exert their activity by binding to nuclear receptors, there is intense interest in developing new ligands for the two known estrogen receptor subtypes, ER-α and ER-β. Characterization assays used to profile new estrogen receptor ligands often utilize receptors from different species, with the assumption that they behave identically. To test this belief, we have profiled a no. of estrogens, other steroids, phytoestrogens and selective estrogen receptor modulators in a solid phase radioligand binding assay using recombinant protein for human, rat, and mouse ER-α and ER-β. Certain compds. show species dependent binding preferences for ER-α or ER-β, leading to differences in receptor subtype selectivity. The amino acids identified by crystallog. as lining the ligand binding cavity are the same among the three species, suggesting that as yet unidentified amino acids contribute to the structure of the binding site. We conclude from this anal. that the ability of a compd. to selectively bind to a particular ER subtype can be species dependent.
- 24Mohler, M. L.; Narayanan, R.; Coss, C. C.; Hu, K.; He, Y.; Wu, Z.; Hong, S.-S.; Hwang, D. J.; Miller, D. D.; Dalton, J. T. Estrogen Receptor β Selective Nonsteroidal Estrogens: Seeking Clinical Indications. Expert Opin. Ther. Pat. 2010, 20, 507– 534, DOI: 10.1517/1354377100365716424https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXjslamu7w%253D&md5=43641194b70be56c763fdf4d8e836e88Estrogen receptor β selective nonsteroidal estrogens: seeking clinical indicationsMohler, Michael L.; Narayanan, Ramesh; Coss, Christopher C.; Hu, Kejiang; He, Yali; Wu, Zhongzhi; Hong, Seoung-Soo; Hwang, Dong Jin; Miller, Duane D.; Dalton, James T.Expert Opinion on Therapeutic Patents (2010), 20 (4), 507-534CODEN: EOTPEG; ISSN:1354-3776. (Informa Healthcare)A review. Importance of the field: Nonsteroidal estrogens have been known since the 1930s. However, the relatively recent (1996) discovery of estrogen receptor subtype β (ERβ) suggested a possible paradigm shift away from SERM-like selectivity. Selective ERβ agonism would potentially allow expansion of estrogenic targeting into new indications (discussed herein) currently precluded by the thrombogenic and hyperproliferative effects of nonselective estrogens. Areas covered in this review: ERβ agonist design has been very successful. Pharmacophores for ERβ selective nonsteroidal estrogens are generally diphenolic compds. that achieve an inter-phenolic distance and geometry similar to 17β-estradiol with few restraints on the nature of the element linking the phenols (or phenol mimetics). The tremendously chemodiverse ERβ agonist patent literature is reviewed, segregating the agonists into structurally similar compds. based on their interphenolic linking elements. What the reader will gain: A comprehensive understanding of the chemotype landscape of this field and an assessment of its maturation. Take home message: Subtype selective ERβ agonist therapy seems very promising. However, more clin. testing is needed to firmly establish its therapeutic potential. At this point, ERβ is a promising target in search of an indication.
- 25Malamas, M. S.; Manas, E. S.; McDevitt, R. E.; Gunawan, I.; Xu, Z. B.; Collini, M. D.; Miller, C. P.; Dinh, T.; Henderson, R. A.; Keith, J. C.; Harris, H. A. Design and Synthesis of Aryl Diphenolic Azoles as Potent and Selective Estrogen Receptor-β Ligands. J. Med. Chem. 2004, 47, 5021– 5040, DOI: 10.1021/jm049719y25https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXnt12gtrc%253D&md5=01c7d1d0fb71105c1adffeb5f0bc7d3bDesign and Synthesis of Aryl Diphenolic Azoles as Potent and Selective Estrogen Receptor-β LigandsMalamas, Michael S.; Manas, Eric S.; McDevitt, Robert E.; Gunawan, Iwan; Xu, Zhang B.; Collini, Michael D.; Miller, Chris P.; Dinh, Tam; Henderson, Ruth A.; Keith, James C., Jr.; Harris, Heather A.Journal of Medicinal Chemistry (2004), 47 (21), 5021-5040CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)New diphenolic azoles as highly selective estrogen receptor-β agonists are reported. The more potent and selective analogs of these series have comparable binding affinities for ERβ as the natural ligand 17β-estradiol but are >100-fold selective over ERα. The design strategy not only followed a traditional SAR approach but also was supported by X-ray structures of ERβ cocrystd. with various ligands as well as mol. modeling studies. These strategies enabled the use of a single conservative residue substitution in the ligand-binding pocket, ERα Met421 → ERβ Ile373, to optimize ERβ selectivity. The 7-position-substituted benzoxazoles were the most selective ligands of both azole series, with ERB-041 (I) being >200-fold selective for ERβ. The majority of ERβ selective agonists tested that were at least ∼50-fold selective displayed a consistent in vivo profile: they were inactive in several models of classic estrogen action (uterotrophic, osteopenia, and vasomotor instability models) and yet were active in the HLA-B27 transgenic rat model of inflammatory bowel disease. These data suggest that ERβ-selective agonists are devoid of classic estrogenic effects and may offer a novel therapy to treat certain inflammatory conditions.
- 26Roman-Blas, J. A.; Castañeda, S.; Cutolo, M.; Herrero-Beaumont, G. Efficacy and Safety of a Selective Estrogen Receptor β Agonist, ERB-041, in Patients with Rheumatoid Arthritis: A 12-Week, Randomized, Placebo-Controlled, Phase II Study. Arthritis Care Res. 2010, 62, 1588– 1593, DOI: 10.1002/acr.2027526https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXhsF2gu7rF&md5=b3b3d9966e0c841340d2077af7cb43e7Efficacy and safety of a selective estrogen receptor β agonist, ERB-041, in patients with rheumatoid arthritis: a 12-week, randomized, placebo-controlled, phase II studyRoman-Blas, Jorge A.; Castaneda, Santos; Cutolo, Maurizio; Herrero-Beaumont, GabrielArthritis Care & Research (2010), 62 (11), 1588-1593CODEN: ARCREG; ISSN:2151-464X. (John Wiley & Sons, Inc.)Objective. Selective estrogen receptor β (ERβ) agonists have demonstrated relevant antiinflammatory effects in different animal models. This study aimed to compare the efficacy and safety of one of these agonists, ERB-041, in subjects with rheumatoid arthritis (RA). Methods. A total of 291 patients with active RA receiving stable doses of methotrexate were randomized to receive 5, 25, or 75 mg of ERB-041 or placebo for 12 wk. The primary end point was the American College of Rheumatol. 20% improvement criteria (ACR20) at 12 wk. Secondary end points included the ACR 50% improvement criteria (ACR50) and the ACR 70% improvement criteria (ACR70) responses, health outcomes measures, C-reactive protein (CRP) levels, and potential exposure-response relationships. Medical history, phys. examn., and lab. values were obtained at screening, baseline, and weeks 2, 4, 8, and 12. Results. No statistically significant difference for the ACR20 was found between the ERB-041 treatment and placebo groups (P = 0.518). Nor was a significant difference obsd. for ACR50 and ACR70 responses, health outcomes measures, CRP levels, and overall incidence of adverse events among all groups. Forty-four subjects (15.1%) discontinued the study and the rate of discontinuation was similar among the treatment groups. The most commonly reported treatment-emergent adverse events were headache (7.6%), nausea (6.2%), infection (4.8%), and bronchitis (4.1%). None of the adverse events was considered treatment related. Conclusion. Although well tolerated and safe, ERB-041 failed to demonstrate antiinflammatory efficacy in RA patients, despite evidence of strong activity in preclin. arthritis models. These results suggest that selective ERβ agonists would not have effects on regulating inflammatory response in RA. Nevertheless, further studies are warranted to establish their efficacy in inflammatory arthritis.
- 27Norman, B. H.; Dodge, J. A.; Richardson, T. I.; Borromeo, P. S.; Lugar, C. W.; Jones, S. A.; Chen, K.; Wang, Y.; Durst, G. L.; Barr, R. J.; Montrose-Rafizadeh, C.; Osborne, H. E.; Amos, R. M.; Guo, S.; Boodhoo, A.; Krishnan, V. Benzopyrans Are Selective Estrogen Receptor β Agonists with Novel Activity in Models of Benign Prostatic Hyperplasia. J. Med. Chem. 2006, 49, 6155– 6157, DOI: 10.1021/jm060491j27https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD28XptFaitr8%253D&md5=8d57f3742a46fe2b1ff176186a6f5d08Benzopyrans Are Selective Estrogen Receptor β Agonists with Novel Activity in Models of Benign Prostatic HyperplasiaNorman, Bryan H.; Dodge, Jeffrey A.; Richardson, Timothy I.; Borromeo, Peter S.; Lugar, Charles W.; Jones, Scott A.; Chen, Keyue; Wang, Yong; Durst, Gregory L.; Barr, Robert J.; Montrose-Rafizadeh, Chahrzad; Osborne, Harold E.; Amos, Robert M.; Guo, Sherry; Boodhoo, Amechand; Krishnan, VenkateshJournal of Medicinal Chemistry (2006), 49 (21), 6155-6157CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)Benzopyran selective estrogen receptor beta agonist-1 (SERBA-1) shows potent, selective binding and agonist function in estrogen receptor β (ERβ) in vitro assays. X-ray crystal structures of SERBA-1 in ERα and β help explain obsd. β-selectivity of this ligand. SERBA-1 in vivo demonstrates involution of the ventral prostate in CD-1 mice (ERβ effect), while having no effect on gonadal hormone levels (ERα effect) at 10× the efficacious dose, consistent with in vitro properties of this mol.
- 28Roehrborn, C. G.; Spann, M. E.; Myers, S. L.; Serviss, C. R.; Hu, L.; Jin, Y. Estrogen Receptor Beta Agonist LY500307 Fails to Improve Symptoms in Men with Enlarged Prostate Secondary to Benign Prostatic Hypertrophy. Prostate Cancer Prostatic Dis. 2015, 18, 43– 48, DOI: 10.1038/pcan.2014.4328https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXhvVOnt73I&md5=a6fa7058d61630f48df3b35c5d637c70Estrogen receptor beta agonist LY500307 fails to improve symptoms in men with enlarged prostate secondary to benign prostatic hypertrophyRoehrborn, C. G.; Spann, M. E.; Myers, S. L.; Serviss, C. R.; Hu, L.; Jin, Y.Prostate Cancer and Prostatic Diseases (2015), 18 (1), 43-48CODEN: PCPDFW; ISSN:1365-7852. (Nature Publishing Group)Background: To assess the efficacy and safety of LY500307, a selective estrogen receptor beta agonist, on lower urinary tract symptoms (LUTS) in patients with enlarged prostate secondary to BPH. Methods: In a randomized, double-blind, placebo-controlled, parallel phase 2, efficacy and safety study, eligible patients with moderate to severe LUTS and prostatic enlargement (≥30 mL) were randomized to placebo or LY500307 at 1, 3, 10 and 25 mg once daily for 24 wk. Primary efficacy end point was change in total International Prostate Symptoms Score (IPSS) after 24 wk. Secondary end points included changes in total prostate vol. (TPV) that served as a proof of concept end point, as well as IPSS quality of life, max. peak urine flow rate (Qmax) and PSA and safety (adverse events, lab. test). Results: A total of 414 patients were randomized when the study was terminated because of insufficient TPV redn., based on a priori defined interim anal. The IPSS mean change from baseline to end point was -3.4±6.8 in the placebo group and -1.3±6.6, -2.6±7.0, -3.7±6.7 and -4.4±5.7 in the 1, 3, 10 and 25 mg LY500307-treated groups, resp. (P>0.05). Similarly, no treatment effect was obsd. for any of the secondary efficacy measures. Incidence of adverse events was comparable between treatment groups, and no clin. meaningful changes in lab. tests were obsd. Conclusions: LY500307 was well tolerated in BPH patients with LUTS at doses up to 25 mg once daily for 24 wk. The study was terminated early because of inadequate efficacy.
- 29Breier, A. The Efficacy and Safety of a Selective Estrogen Receptor Beta Agonist (LY500307) for Negative Symptoms and Cognitive Impairment Associated With Schizophrenia—Full Text View—ClinicalTrials.gov. https://clinicaltrials.gov/ct2/show/NCT01874756 (accessed Apr 27, 2020).There is no corresponding record for this reference.
- 30Effects of an ER Beta Agonist (Lilly Compound LY500307) on Estradiol-Withdrawal-Induced Mood Symptoms in Women with Past Perimenopausal Depression—Tabular View—ClinicalTrials.gov. https://clinicaltrials.gov/ct2/show/record/NCT03689543 (accessed Apr 27, 2020).There is no corresponding record for this reference.
- 31Paterni, I.; Bertini, S.; Granchi, C.; Macchia, M.; Minutolo, F. Estrogen Receptor Ligands: A Patent Review Update. Expert Opin. Ther. Pat. 2013, 23, 1247– 1271, DOI: 10.1517/13543776.2013.80520631https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhsVyns7fO&md5=6f747f7fccf0953fc64cd61b5f287dd6Estrogen receptor ligands: a patent review updatePaterni, Ilaria; Bertini, Simone; Granchi, Carlotta; Macchia, Marco; Minutolo, FilippoExpert Opinion on Therapeutic Patents (2013), 23 (10), 1247-1271CODEN: EOTPEG; ISSN:1354-3776. (Informa Healthcare)A review. Introduction: The role of estrogens is mostly mediated by two nuclear receptors (ERα and ERβ) and a membrane-assocd. G-protein (GPR30 or GPER), and it is not limited to reprodn., but it extends to the skeletal, cardiovascular and central nervous systems. Various pathologies such as cancer, inflammatory, neurodegenerative and metabolic diseases are often assocd. with dysfunctions of the estrogenic system. Therapeutic interventions by agents that affect the estrogenic signaling pathway might be useful in the treatment of many dissimilar diseases. Areas covered: The massive chemodiversity of ER ligands, limited to patented small mols., is herein reviewed. The reported compds. are classified on the basis of their chem. structures. Non-steroidal derivs., which mostly consist of diphenolic compds., are further segregated into chem. classes based on their central scaffold. Expert opinion: Estrogens have been used for almost a century and their earlier applications have concerned interventions in the female reproductive functions, as well as the treatment of some estrogen-dependent cancers and osteoporosis. Since the discovery of ERβ in 1996, the patent literature has started to pay a progressively increasing attention to this newer receptor subtype, which holds promise as a target for new indications, most of which still need to be clin. validated.
- 32Endo, Y.; Iijima, T.; Yamakoshi, Y.; Yamaguchi, M.; Fukasawa, H.; Shudo, K. Potent Estrogenic Agonists Bearing Dicarba-closo-Dodecaborane as a Hydrophobic Pharmacophore. J. Med. Chem. 1999, 42, 1501– 1504, DOI: 10.1021/jm990072532https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXisVaqsbY%253D&md5=b487b1fcfa55e50d83a5cf96d8b357faPotent Estrogenic Agonists Bearing Dicarba-closo-dodecaborane as a Hydrophobic PharmacophoreEndo, Yasuyuki; Iijima, Toru; Yamakoshi, Yuko; Yamaguchi, Mitsuhiro; Fukasawa, Hiroshi; Shudo, KoichiJournal of Medicinal Chemistry (1999), 42 (9), 1501-1504CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)The novel carborane contg. mols., e.g. 1-(4-hydroxyphenyl)-1,12-dicarba-closo-dodecaborane, with potent estrogenic activity was developed. The unique character of biol. active mols. contg. a carborane skeleton may give to unusual membrane transport characteristics and metab., compared with conventional active mols. The superagonistic properties of the carborane-contg. compds. raise the possibility that structure-function studies could lead to the development of more selective estrogen agonists and antagonists, which could be useful as therapeutic agents for a wide variety of conditions.
- 33Barth, R. F.; Zhang, Z.; Liu, T. A Realistic Appraisal of Boron Neutron Capture Therapy as a Cancer Treatment Modality. Cancer Commun. 2018, 38, 36 DOI: 10.1186/s40880-018-0280-533https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC1MbosFamuw%253D%253D&md5=aaf7960e10efcae0cbdcaa5113ba2bdaA realistic appraisal of boron neutron capture therapy as a cancer treatment modalityBarth Rolf F; Zhang Zizhu; Liu TongCancer communications (London, England) (2018), 38 (1), 36 ISSN:.Boron neutron capture therapy (BNCT) is a binary therapeutic modality based on the nuclear capture and fission reactions that occur when the stable isotope boron-10 is irradiated with neutrons to produce high-energy alpha particles and recoiling lithium-7 nuclei. In this Commentary we will focus on a number of papers that were presented at a Symposium entitled "Current Clinical Status of Boron Neutron Capture Therapy and Paths to the Future", which was held in September 2017 at the China National Convention Center in Beijing. Results were presented by clinicians from Japan, Finland, the United States, the China mainland and Taiwan, China who have been working in the multiple disciplines that are required for carrying out clinical BNCT. The main focus was on the treatment of patients with malignant brain tumors, recurrent tumors of the head and neck region, and cutaneous melanomas. The results obtained in treating these patients were reported in detail and, although most of the patients with brain tumors and head and neck cancer were not cured, there was evidence of some clinical efficacy. Although there are a number of problems that must be addressed, further clinical studies to evaluate the efficacy of BNCT are warranted. First, despite considerable effort by numerous investigators over the past 40 years, there still are only two boron-containing drugs in clinical use, L-boronophenylalanine (BPA) and sodium borocaptate (BSH). Therefore, until new and more effective boron delivery agents are developed, efforts should be directed to improving the dosing and delivery of BPA and BSH. Second, due to a variety of reasons, nuclear reactor-based BNCT has ended except for its use in the China mainland and Taiwan. Therefore, the future of BNCT depends upon the results of the ongoing Phase II clinical trials that are being carried out in Japan and the soon to be initiated trials that will be carried out in Finland. If the results obtained from these clinical trials are sufficiently promising, then BNCT will have a clear path to the future, especially for patients with the therapeutically challenging malignancies that in the past have been treated with reactor-based BNCT.
- 34Soloway, A. H.; Tjarks, W.; Barnum, B. A.; Rong, F.-G.; Barth, R. F.; Codogni, I. M.; Wilson, J. G. The Chemistry of Neutron Capture Therapy. Chem. Rev. 1998, 98, 1515– 1562, DOI: 10.1021/cr941195u34https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1cXjtlygtL8%253D&md5=52bcd4d4e2f37833d76092e9a1867b3cThe Chemistry of Neutron Capture TherapySoloway, Albert H.; Tjarks, Werner; Barnum, Beverly A.; Rong, Feng-Guang; Barth, Rolf F.; Codogni, Iwona M.; Wilson, J. GeraldChemical Reviews (Washington, D. C.) (1998), 98 (4), 1515-1562CODEN: CHREAY; ISSN:0009-2665. (American Chemical Society)A review with 633 refs.
- 35Soloway, A. H.; Wright, R. L.; Messer, J. R. Evaluation of Boron Compounds for Use in Neutron Capture Therapy of Brain Tumors. I. Animal Investigations. J. Pharmacol. Exp. Ther. 1961, 134, 117– 12235https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaF38Xks1al&md5=e291c2430bd5f235a4b0f6e3a429a7d3Evaluation of boron compounds for use in neutron-capture therapy of brain tumors. I. Animal investigationsSoloway, A. H.; Wright, R. L.; Messer, J. R.Journal of Pharmacology and Experimental Therapeutics (1961), 134 (), 117-22CODEN: JPETAB; ISSN:0022-3565.cf. CA 54, 1374d.-m-Boronosuccinanilic, 3-amino-4-carboxybenzeneboronic, m- and p-carboxybenzeneboronic, 2-nitrobenzene-l,4-diboronic, o-(2-carboxyacetamidoethyl)benzeneboronic, 2-acetamidobenzeneboronic, and m-ureidobenzeneboronic acids, p-boronophenylalanine, Na perhydrodecaborate, and boric acid were evaluated in mice with transplanted glioma (ependymoma) for toxicity. The tumor-to-brain ratios and the concns. in various organ tissues were detd. The 1st 2 compds. appeared promising and were studied further in cats by intravenous and intracarotid artery injection.
- 36Stockmann, P.; Gozzi, M.; Kuhnert, R.; Sárosi, M. B.; Hey-Hawkins, E. New Keys for Old Locks: Carborane-Containing Drugs as Platforms for Mechanism-Based Therapies. Chem. Soc. Rev. 2019, 48, 3497– 3512, DOI: 10.1039/C9CS00197B36https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXhtF2nt7vI&md5=b2a1f1247161aeedf257cd807d2a30fdNew keys for old locks: carborane-containing drugs as platforms for mechanism-based therapiesStockmann, Philipp; Gozzi, Marta; Kuhnert, Robert; Sarosi, Menyhart B.; Hey-Hawkins, EvamarieChemical Society Reviews (2019), 48 (13), 3497-3512CODEN: CSRVBR; ISSN:0306-0012. (Royal Society of Chemistry)A review. Icosahedral carboranes in medicine are still an emerging class of compds. with potential beneficial applications in drug design. These highly hydrophobic clusters are potential "new keys for old locks" which open up an exciting field of research for well-known, but challenging important therapeutic substrates, as demonstrated by the numerous examples discussed in this review.
- 37Leśnikowski, Z. J. Challenges and Opportunities for the Application of Boron Clusters in Drug Design. J. Med. Chem. 2016, 59, 7738– 7758, DOI: 10.1021/acs.jmedchem.5b0193237https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC28XmvFSgs7c%253D&md5=1d4c3720cdc649c5fc3de397bea15e83Challenges and Opportunities for the Application of Boron Clusters in Drug DesignLesnikowski, Zbigniew J.Journal of Medicinal Chemistry (2016), 59 (17), 7738-7758CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)There are two branches in boron medicinal chem.: the first focuses on single boron atom compds., and the second utilizes boron clusters. Boron clusters and their heteroatom counterparts belong to the family of cage compds. A subset of this extensive class of compds. includes dicarbadodecaboranes, which have the general formula C2B10H12, and their metal biscarboranyl complexes, metallacarboranes, with the formula [M(C2B10H12)2-2]. The unique properties of boron clusters have resulted in their utilization in applications such as in pharmacophores, as scaffolds in mol. construction, and as modulators of bioactive compds. This Perspective presents an overview of the properties of boron clusters that are pertinent for drug discovery, recent applications in the design of various classes of drugs, and the potential use of boron clusters in the construction of new pharmaceuticals.
- 38Leśnikowski, Z. J. What Are the Current Challenges with the Application of Boron Clusters to Drug Design?. Expert Opin. Drug Discovery 2020, 481, DOI: 10.1080/17460441.2021.1867531There is no corresponding record for this reference.
- 39Sedlák, D.; Eignerová, B.; Dračínský, M.; Janoušek, Z.; Bartůněk, P.; Kotora, M. Synthesis and Evaluation of 17α-(Carboranylalkyl)Estradiols as Ligands for Estrogen Receptors α and β. J. Organomet. Chem. 2013, 747, 178– 183, DOI: 10.1016/j.jorganchem.2013.06.01339https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhtVKgsrrL&md5=30ff55174dd93879e3cdd196e1032c6aSynthesis and evaluation of 17α-(carboranylalkyl)estradiols as ligands for estrogen receptors α and βSedlak, David; Eignerova, Barbara; Dracinsky, Martin; Janousek, Zbynek; Bartunek, Petr; Kotora, MartinJournal of Organometallic Chemistry (2013), 747 (), 178-183CODEN: JORCAI; ISSN:0022-328X. (Elsevier B.V.)A series of 17α-(carboranylalkyl)estradiols was synthesized using cross-metathesis reaction of 17α-allyl- and 17α-vinylestradiols with allylcarboranes catalyzed by Hoveyda-Grubbs 2nd generation catalyst. The prepd. estradiol derivs. were tested in the panel of cell-based reporter assays including all steroid receptors. The compds. mainly activated estrogen receptors α and β and tended to be weakly selective for ERα. Besides ERα and ERβ, we have also detected a weak agonistic activity on AR and PR at micromolar concns. We didn't observe any antagonistic effect with the exception of two receptors: GR and MR which were inhibited with some of the tested ligands. However, the inhibition was detectable at concns. exceeding by far those needed to activate estrogen receptors.
- 40Eignerová, B.; Sedlák, D.; Dračínský, M.; Bartůněk, P.; Kotora, M. Synthesis and Biochemical Characterization of a Series of 17α-Perfluoroalkylated Estradiols as Selective Ligands for Estrogen Receptor α. J. Med. Chem. 2010, 53, 6947– 6953, DOI: 10.1021/jm100563h40https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXhtFWlsb7E&md5=be45ac8fd4423d38ba410b2e080676faSynthesis and Biochemical Characterization of a Series of 17α-Perfluoroalkylated Estradiols as Selective Ligands for Estrogen Receptor αEignerova, Barbara; Sedlak, David; Dracinsky, Martin; Bartunek, Petr; Kotora, MartinJournal of Medicinal Chemistry (2010), 53 (19), 6947-6953CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)Despite intensive research efforts, the distinct biol. roles of two closely related estrogen receptors, ERα and ERβ, are only partially understood. Therefore, ligands selective for either of two isotypes are useful research tools because they allow for exerting a desired subset of biol. effects mediated by only one of the receptors. Here we report on the synthesis of a new class of potent and selective ligands for ERα represented by a series of 17α-substituted estradiols bearing lipophilic perfluoroalkyl chains. These 17α-perfluoroalkylated estradiols were synthesized by Ru-catalyzed cross metathesis reactions of 17α-allyl- or 17α-vinylestradiols with perfluoroalkylpropenes. Compds. were tested in both agonistic and antagonistic modes using a panel of stable steroid receptor reporter cell lines established in U2OS cells and consisting of ERα-LBD, ERβ-LBD, GR-LBD, and MR-LBD reporters. Some of the compds. are potent and selective agonists of ERα, exhibiting weak partial to no detectable agonistic activity on ERβ. Notably, I is the most ERα selective ligand of the prepd. compds. because it activates ERα but inhibits ERβ. In addn., some compds. are pure agonists on ERα but show mixed agonistic/antagonistic profile on ERβ which is a typical pattern obsd. for selective estrogen receptor modulators (SERMs).
- 41Sedlák, D.; Novák, P.; Kotora, M.; Bartůněk, P. Synthesis and Evaluation of 17α-Arylestradiols as Ligands for Estrogen Receptor α and β. J. Med. Chem. 2010, 53, 4290– 4294, DOI: 10.1021/jm901898a41https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXltVyhuro%253D&md5=f1c7acaa44d748d6c5adc341290a79cbSynthesis and Evaluation of 17α-Arylestradiols as Ligands for Estrogen Receptor α and βSedlak, David; Novak, Petr; Kotora, Martin; Bartunek, PetrJournal of Medicinal Chemistry (2010), 53 (10), 4290-4294CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)To identify novel estrogen receptor ligands a series of substituted 17α-arylestradiols I [X = C(CO2Et)2, C(COMe)2, C(CO2Et)COMe, C(CO2Et)CN, NTs, O] were synthesized using the catalytic [2 + 2 + 2]-cyclotrimerization of 17α-ethynylestradiol with various 1,7-diynes HC≡CCH2XCH2C≡CH in the presence of Wilkinson's catalysts [Rh(PPh3)3Cl]. The compds. were subjected to competitive binding assays, cell-based luciferase reporter assays, and proliferation assays. These expts. confirmed their estrogenic character and revealed some interesting properties like mixed partial/full agonism for ERα/ERβ and different selectivity as a result of differing potencies for either ER.
- 42Endo, Y.; Yamamoto, K.; Kagechika, H. Utility of Boron Clusters for Drug Design. Relation between Estrogen Receptor Binding Affinity and Hydrophobicity of Phenols Bearing Various Types of Carboranyl Groups. Bioorg. Med. Chem. Lett. 2003, 13, 4089– 4092, DOI: 10.1016/j.bmcl.2003.08.03942https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3sXosVWrtrk%253D&md5=337630c26a43eb82c590ebb7b6cd8e15Utility of boron clusters for drug design. Relation between estrogen receptor binding affinity and hydrophobicity of phenols bearing various types of carboranyl groupsEndo, Yasuyuki; Yamamoto, Keisuke; Kagechika, HiroyukiBioorganic & Medicinal Chemistry Letters (2003), 13 (22), 4089-4092CODEN: BMCLE8; ISSN:0960-894X. (Elsevier Science B.V.)High binding affinity for estrogen receptor and the appearance of estrogenic activity require a phenolic ring and an appropriate hydrophobic group adjacent to the phenolic ring. A quant. structure-activity relationship anal. based on the values of logP and the pKa of the phenolic group showed that the hydrophobicity of these compds. is highly correlated to the estrogen receptor α (ERα)-binding affinity. These results should be useful for application of these spherical boron clusters (dicarba-closo-dodecaboranes; carboranes) as hydrophobic pharmacophores in drug design, as well as for microscopic anal. of ER-ligand interactions.
- 43Endo, Y.; Iijima, T.; Yamakoshi, Y.; Fukasawa, H.; Miyaura, C.; Inada, M.; Kubo, A.; Itai, A. Potent Estrogen Agonists Based on Carborane as a Hydrophobic Skeletal Structure: A New Medicinal Application of Boron Clusters. Chem. Biol. 2001, 8, 341– 355, DOI: 10.1016/S1074-5521(01)00016-343https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3MXjs1ahsr4%253D&md5=1f1af2e8526772e69d711f538fc1d82dPotent estrogen agonists based on carborane as a hydrophobic skeletal structure: a new medicinal application of boron clustersEndo, Yasuyuki; Iijima, Toru; Yamakoshi, Yuko; Fukasawa, Hiroshi; Miyaura, Chisato; Inada, Masaki; Kubo, Asako; Itai, AkikoChemistry & Biology (2001), 8 (4), 341-355CODEN: CBOLE2; ISSN:1074-5521. (Elsevier Science Ltd.)Background: Carboranes (dicarba-closo-dodecaboranes) are a class of carbon-contg. polyhedral boron-cluster compds. having remarkable thermal stability and exceptional hydrophobicity. Applications of the unique structural and chem. properties offered by icosahedral carboranes in boron neutron capture therapy have received increasing attention over the past 30 yr. However, these features of carboranes may allow another application as a hydrophobic pharmacophore in biol. active mols. that interact hydrophobically with receptors. Results: We have designed candidate estrogen-receptor-binding compds. having carborane as a hydrophobic skeletal structure and synthesized them. The most potent compd. bearing a carborane cage exhibited activity at least 10-fold greater than that of 17β-estradiol in the luciferase reporter gene assay. Estrogen receptor-α-binding data for the compd. were consistent with the results of the luciferase reporter gene assay. The compd. also showed potent in vivo effects on the recovery of uterine wt. and bone loss in ovariectomized mice. Conclusion: Further development of the potent carborane-contg. estrogenic agonists described here, having a new skeletal structure and unique characteristics, should yield novel therapeutic agents, esp. selective estrogen receptor modulators. Furthermore, the suitability of the spherical carborane cage for binding to the cavity of the estrogen receptor-α ligand-binding domain should provide a basis for a similar approach to developing novel ligands for other steroid receptors.
- 44Ohta, K.; Ogawa, T.; Oda, A.; Kaise, A.; Endo, Y. Design and Synthesis of Carborane-Containing Estrogen Receptor-Beta (ERβ)-Selective Ligands. Bioorg. Med. Chem. Lett. 2015, 25, 4174– 4178, DOI: 10.1016/j.bmcl.2015.08.00744https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2MXhtlSlsLjF&md5=18ce9668981a0f26829a981d79f8addbDesign and synthesis of carborane-containing estrogen receptor-beta (ERβ)-selective ligandsOhta, Kiminori; Ogawa, Takumi; Oda, Akifumi; Kaise, Asako; Endo, YasuyukiBioorganic & Medicinal Chemistry Letters (2015), 25 (19), 4174-4178CODEN: BMCLE8; ISSN:0960-894X. (Elsevier B.V.)Candidates for highly selective estrogen receptor-beta (ERβ) ligands were designed and synthesized based on carborane-contg. ER ligands as lead compds. Among them, p-carboranylcyclohexanol derivs. 8a and 8b exhibited high ERβ selectivity in competitive binding assay: for example, 8a showed 56-fold selectivity for ERβ over ERα. Docking studies of 8a and 8b with the ERα and ERβ ligand-binding domains (LBDs) suggested that the p-carborane cage of the ligands is located close to key amino acid residues that influence ER-subtype selectivity, i.e., Leu384 in the ERα LBD and Met336 in the ERβ LBD. The p-carborane cage in 8a and 8b appears to play a crucial role in the increased ERβ selectivity.
- 45Coult, R.; Fox, M. A.; Gill, W. R.; Herbertson, P. L.; MacBride, J. A. H.; Wade, K. C-Arylation and C-Heteroarylation of Icosahedral Carboranes via Their Copper(I) Derivatives. J. Organomet. Chem. 1993, 462, 19– 29, DOI: 10.1016/0022-328X(93)83337-U45https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK2cXitFOktbc%253D&md5=cbd713095584fc4dbaf5b88cee325275C-arylation and C-heteroarylation of icosahedral carboranes via their copper(I) derivativesCoult, Robert; Fox, Mark A.; Gill, Wendy R.; Herbertson, Penelope L.; MacBride, J. A. Hugh; Wade, KennethJournal of Organometallic Chemistry (1993), 462 (1-2), 19-29CODEN: JORCAI; ISSN:0022-328X.Reaction between C-mono- or C,C'-di-copper(I) derivs. of 1,2-, 1,7-, or 1,12-dicarba-closo-dodecaborane(12) and aryl iodides in the presence of pyridine gives the corresponding C-mono- or C,C'-diaryl derivs. of 1,7- and 1,12-dicarba-closo-dodecaboranes(12); 1,2-dicarba-closo-dodecaborane(12) gives only the C-monoaryl product. Cyclic or linear arylene coupled systems are obtained when di-iodoarenes are used. Copper(I) derivs. may be generated from C-unsubstituted or C-monosubstituted carboranes using copper(I) t-butoxide when substituents incompatible with the use of C-lithio-intermediates are involved. The C-copper(I) deriv. of 1,2-dicarba-closo-dodecaborane(12) gives 1,2-di-2'-pyridyl-1,2-dicarba-closo-dodecaborane(12) specifically with 2-bromopyridine. The (inferred) intermediate mono-2-pyridyl-deriv., obtained independently from 2-ethynylpyridine and the di-Me sulfide complex of decaborane, gives 1-phenyl-2,2'-pyridyl-1,2-dicarba-closo-dodecaborane(12) upon conversion into its copper(I) deriv. and treatment with iododobenzene. However, the copper(I) deriv. of 1-phenyl-1,2-dicarba-closo-dodecaborane(12) does not react to a significant extent with 2-bromopyridine.
- 46Corey, E. J.; Bakshi, R. K.; Shibata, S.; Chen, C. P.; Singh, V. K. A Stable and Easily Prepared Catalyst for the Enantioselective Reduction of Ketones. Applications to Multistep Syntheses. J. Am. Chem. Soc. 1987, 109, 7925– 7926, DOI: 10.1021/ja00259a07546https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaL1cXnsFShtA%253D%253D&md5=4deaae8502b4e75f0744e46a9e774decA stable and easily prepared catalyst for the enantioselective reduction of ketones. Applications to multistep synthesesCorey, E. J.; Bakshi, Raman K.; Shibata, Saizo; Chen, Chung Pin; Singh, Vinod K.Journal of the American Chemical Society (1987), 109 (25), 7925-6CODEN: JACSAT; ISSN:0002-7863.The readily available and stable oxazaborolidine I is an excellent catalyst for the enantioselective redn. of a wide range of ketones. The stereochem. course of the redn. is predictable and the secondary alcs. formed are generally produced with high enantioselectivity. Use of I or its enantiomer provides outstanding control of stereochem. at C(15) in prostaglandin synthesis. A general and highly enantioselective synthesis of trans-2,5-diarylfurans, potent antagonists of platelet activating factor, also illustrates the value of this enantioselective redn. Both I and its enantiomer can be obtained from (±)-pyroglutamic acid.
- 47Dale, J. A.; Mosher, H. S. Nuclear Magnetic Resonance Enantiomer Regents. Configurational Correlations via Nuclear Magnetic Resonance Chemical Shifts of Diastereomeric Mandelate, O-Methylmandelate, and.Alpha.-Methoxy-.Alpha.-Trifluoromethylphenylacetate (MTPA) Esters. J. Am. Chem. Soc. 1973, 95, 512– 519, DOI: 10.1021/ja00783a03447https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaE3sXotlOjtw%253D%253D&md5=27a7f9c81b89c6b29ffed705dc4f689cNuclear magnetic resonance enantiomer regents. Configurational correlations via nuclear magnetic resonance chemical shifts of diastereomeric mandelate, O-methylmandelate, and α-methoxy-α-trifluoromethylphenylacetate (MTPA) estersDale, James A.; Mosher, Harry S.Journal of the American Chemical Society (1973), 95 (2), 512-19CODEN: JACSAT; ISSN:0002-7863.An empirically derived correlation of configuration and NMR chem. shifts for diastereomeric mandelate, O-methylmandelate and α-methoxy-α-trifluoromethylphenylacetate esters has been developed and rationalized in terms of useful models. These models have been successfully applied to over 40 examples. The correlations involve the relative chem. shifts of the proton resonances from the groups attached to the carbinyl carbon of these diastereomeric esters.
- 48Hoye, T. R.; Jeffrey, C. S.; Shao, F. Mosher Ester Analysis for the Determination of Absolute Configuration of Stereogenic (Chiral) Carbinol Carbons. Nat. Protoc. 2007, 2, 2451– 2458, DOI: 10.1038/nprot.2007.35448https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXhtFOksL3E&md5=741bb9fd8eb7c18c310196f298e5ea3fMosher ester analysis for the determination of absolute configuration of stereogenic (chiral) carbinol carbonsHoye, Thomas R.; Jeffrey, Christopher S.; Shao, FengNature Protocols (2007), 2 (10), 2451-2458CODEN: NPARDW; ISSN:1750-2799. (Nature Publishing Group)This protocol details the most commonly used NMR-based method for deducing the configuration of otherwise unknown stereogenic, secondary carbinol (alc.) centers (R1R2CHOH (or the analogous amines where OH is replaced by NH2)). This 'Mosher ester anal.' relies on the fact that the protons in diastereomeric α-methoxy-α-trifluoromethylphenylacetic acid (MTPA) esters (i.e., those derived from conjugation of the carbinol under interrogation with MTPA) display different arrays of chem. shifts (δs) in their 1H NMR spectra. The protocol consists of the following: (i) prepn. of each of the diastereomeric S- and R-MTPA esters and (ii) comparative (ΔδSR) anal. of the 1H NMR spectral data of these two esters. By analyzing the sign of the difference in chem. shifts for a no. of analogous pairs of protons (the set of ΔδSR values) in the diastereomeric esters (or amides), the abs. configuration of the original carbinol (or amino) stereocenter can be reliably deduced. A typical Mosher ester anal. requires approx. 4-6 h of active effort over a 1- to 2-d period.
- 49Goto, T.; Ohta, K.; Fujii, S.; Ohta, S.; Endo, Y. Design and Synthesis of Androgen Receptor Full Antagonists Bearing a P-Carborane Cage: Promising Ligands for Anti-Androgen Withdrawal Syndrome. J. Med. Chem. 2010, 53, 4917– 4926, DOI: 10.1021/jm100316f49https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXmvFaktbs%253D&md5=e1df86c4bdb95869f329aac6442419d7Design and Synthesis of Androgen Receptor Full Antagonists Bearing a p-Carborane Cage: Promising Ligands for Anti-Androgen Withdrawal SyndromeGoto, Tokuhito; Ohta, Kiminori; Fujii, Shinya; Ohta, Shigeru; Endo, YasuyukiJournal of Medicinal Chemistry (2010), 53 (13), 4917-4926CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)Pure androgen receptor (AR) full antagonists are candidates to treat anti-androgen refractory prostate cancers. We previously developed a carborane-contg. AR antagonist, 3-(12-hydroxymethyl-1,12-dicarba-closo-dodecaborane-1-yl)benzonitrile (BA341), which was more potent than hydroxyflutamide (4) but acted as an agonist toward LNCaP prostate cancer cells expressing T877A AR mutant. Here, we designed and synthesized novel AR full antagonists, 3-[12-(1-hydroxy-2-Ar-ethyl)-1,12-dicarba-closo-dodecaborane-1-yl]benzonitrile (e.g., Ar = 4-XC6H4O, X = F 7b, MeO, NO2, NHCOMe and Ar = 4-XC6H4S, X = H, F 8b, MeO, NHCOMe, CF3 and Ar = 4-XC6H4SO2) structurally based upon the clin. used AR full antagonist (R)-bicalutamide (5) to test our hypothesis that the carborane cage is suitable as a hydrophobic pharmacophore for AR ligands. 7A and 8b showed good biol. profiles in AR binding and transactivation assays and dose-dependently inhibited the testosterone-induced proliferation of LNCaP cells, as well as SC-3 cells. The IC50 values of compds. 7b and 8b were 3.8 × 10-7 and 4.2 × 10-7 M, resp. [5, 8.7 × 10-7 M]. Since compds. 7b and 8b did not show any agonistic activity in functional assays, they seem to be pure AR full antagonists and are therefore candidates for treatment of anti-androgen withdrawal syndrome.
- 50Ohta, K.; Goto, T.; Fujii, S.; Kawahata, M.; Oda, A.; Ohta, S.; Yamaguchi, K.; Hirono, S.; Endo, Y. Crystal Structure, Docking Study and Structure–Activity Relationship of Carborane-Containing Androgen Receptor Antagonist 3-(12-Hydroxymethyl-1,12-Dicarba-closo-Dodecaboran-1-Yl)Benzonitrile. Bioorg. Med. Chem. 2011, 19, 3540– 3548, DOI: 10.1016/j.bmc.2011.04.01750https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXmvFWmt7Y%253D&md5=ad316a904afbef54577cf99651706340Crystal structure, docking study and structure-activity relationship of carborane-containing androgen receptor antagonist 3-(12-hydroxymethyl-1,12-dicarba-closo-dodecarboran-1-yl)benzonitrileOhta, Kiminori; Goto, Tokuhito; Fujii, Shinya; Kawahata, Masatoshi; Oda, Akifumi; Ohta, Shigeru; Yamaguchi, Kentaro; Hirono, Shuichi; Endo, YasuyukiBioorganic & Medicinal Chemistry (2011), 19 (11), 3540-3548CODEN: BMECEP; ISSN:0968-0896. (Elsevier B.V.)A potent androgen receptor (AR) antagonist, 3-(12-hydroxymethyl-1,12 dicarba-closo-dodecarboran-1-yl)benzonitrile (3a, BA341), contains a p-carborane cage as a hydrophobic pharmacophore. We elucidated the structural properties of 3a by single-crystal x-ray diffraction anal. and conducted a docking study of 3a with hAR LBD. The cyano group of 3a formed hydrogen bonds with Gln711 and Arg752 and the hydroxymethyl group did so with Asn705 and Thr877 in hAR LBD. The bulky p-carborane cage was accommodated in the hydrophobic pocket of hAR LBD. To understand the structure-activity relation around the hydroxymethyl group of 3a, we designed, synthesized, and evaluated the biol. activities of various novel AR ligands. Since the biol. activities of carbonyl compds. 8a, 8b, and 8c were similar to or weaker than those of the parent hydroxyl compds. 3a, 7a, and 7b, it seems to be necessary to have not only a hydrogen bonding acceptor, but also a hydrogen bonding donor adjacent to the hydroxymethyl group of 3a for efficient interaction with hAR LBD.
- 51Anzenbacher, P.; Zanger, U. M., Eds. Metabolism of Drugs and Other Xenobiotics, 1st ed.; John Wiley & Sons, Ltd., 2012.There is no corresponding record for this reference.
- 52Lee, A. J.; Cai, M. X.; Thomas, P. E.; Conney, A. H.; Zhu, B. T. Characterization of the Oxidative Metabolites of 17β-Estradiol and Estrone Formed by 15 Selectively Expressed Human Cytochrome P450 Isoforms. Endocrinology 2003, 144, 3382– 3398, DOI: 10.1210/en.2003-019252https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3sXlslGjsb0%253D&md5=dc77411de262647d6fcf0c28c27d66e7Characterization of the oxidative metabolites of 17β-estradiol and estrone formed by 15 selectively expressed human cytochrome p450 isoformsLee, Anthony J.; Cai, May Xiaoxin; Thomas, Paul E.; Conney, Allan H.; Zhu, Bao TingEndocrinology (2003), 144 (8), 3382-3398CODEN: ENDOAO; ISSN:0013-7227. (Endocrine Society)We systematically characterized the oxidative metabolites of 17β-estradiol and estrone formed by 15 human cytochrome P 450 (CYP) isoforms. CYP1A1 had high activity for 17β-estradiol 2-hydroxylation, followed by 15α-, 6α-, 4-, and 7α-hydroxylation. However, when estrone was the substrate, CYP1A1 formed more 4-hydroxyestrone than 15α- or 6α-hydroxyestrone, with 2-hydroxyestrone as the major metabolite. CYP1A2 had the highest activity for the 2-hydroxylation of both 17β-estradiol and estrone, although it also had considerable activity for their 4-hydroxylation (9-13% of 2-hydroxylation). CYP1B1 mainly catalyzed the formation of catechol estrogens, with 4-hydroxyestrogens predominant. CYP2A6, 2B6, 2C8, 2C9, 2C19, and 2D6 each showed a varying degree of low catalytic activity for estrogen 2-hydroxylation, whereas CYP2C18 and CYP2E1 did not show any detectable estrogen-hydroxylating activity. CYP3A4 had strong activity for the formation of 2-hydroxyestradiol, followed by 4-hydroxyestradiol and an unknown polar metabolite, and small amts. of 16α- and 16β-hydroxyestrogens were also formed. The ratio of 4- to 2-hydroxylation of 17β-estradiol or estrone with CYP3A4 was 0.22 or 0.51, resp. CYP3A5 had similar catalytic activity for the formation of 2- and 4-hydroxyestrogens. Notably, CYP3A5 had an unusually high ratio of 4- to 2-hydroxylation of 17β-estradiol or estrone (0.53 or 1.26, resp.). CYP3A4 and 3A5 also catalyzed the formation of nonpolar estrogen metabolite peaks (chromatog. less polar than estrone). CYP3A7 had a distinct catalytic activity for the 16α-hydroxylation of estrone, but not 17β-estradiol. CYP4A11 had little catalytic activity for the metab. of 17β-estradiol and estrone. In conclusion, many human CYP isoforms are involved in the oxidative metab. of 17β-estradiol and estrone, with a varying degree of catalytic activity and distinct regioselectivity.
- 53Yano, J. K.; Wester, M. R.; Schoch, G. A.; Griffin, K. J.; Stout, C. D.; Johnson, E. F. The Structure of Human Microsomal Cytochrome P450 3A4 Determined by X-ray Crystallography to 2.05-Å Resolution. J. Biol. Chem. 2004, 279, 38091– 38094, DOI: 10.1074/jbc.C40029320053https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXnt1Oqs7c%253D&md5=59a8fa760069d365165ddeeb9e4081d4The Structure of Human Microsomal Cytochrome P450 3A4 Determined by X-ray Crystallography to 2.05-Å ResolutionYano, Jason K.; Wester, Michael R.; Schoch, Guillaume A.; Griffin, Keith J.; Stout, C. David; Johnson, Eric F.Journal of Biological Chemistry (2004), 279 (37), 38091-38094CODEN: JBCHA3; ISSN:0021-9258. (American Society for Biochemistry and Molecular Biology)The structure of P 450 3A4 was detd. by x-ray crystallog. to 2.05-Å resoln. P 450 3A4 catalyzes the metabolic clearance of a large no. of clin. used drugs, and a no. of adverse drug-drug interactions reflect the inhibition or induction of the enzyme. P 450 3A4 exhibits a relatively large substrate-binding cavity that is consistent with its capacity to oxidize bulky substrates such as cyclosporin, statins, taxanes, and macrolide antibiotics. Family 3A P450s also exhibit unusual kinetic characteristics that suggest simultaneous occupancy by smaller substrates. Although the active site vol. is similar to that of P 450 2C8 (PDB code: 1PQ2), the shape of the active site cavity differs considerably due to differences in the folding and packing of portions of the protein that form the cavity. Compared with P 450 2C8, the active site cavity of 3A4 is much larger near the heme iron. The lower constraints on the motions of small substrates near the site of oxygen activation may diminish the efficiency of substrate oxidn., which may in turn be improved by space restrictions imposed by the presence of a second substrate mol. The structure of P 450 3A4 should facilitate a better understanding of the substrate selectivity of the enzyme.
- 54Anzenbacherová, E.; Bec, N.; Anzenbacher, P.; Hudecek, J.; Soucek, P.; Jung, C.; Munro, A. W.; Lange, R. Flexibility and Stability of the Structure of Cytochromes P450 3A4 and BM-3. Eur. J. Biochem. 2000, 267, 2916– 2920, DOI: 10.1046/j.1432-1327.2000.01305.x54https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3cXjs1Gjs7w%253D&md5=9e358cae19770cf66d9ec8cd6ffe595eFlexibility and stability of the structure of cytochromes P450 3A4 and BM-3Anzenbacherova, Eva; Bec, Nicole; Anzenbacher, Pavel; Hudecek, Jiri; Soucek, Pavel; Jung, Christiane; Munro, Andrew W.; Lange, ReinhardEuropean Journal of Biochemistry (2000), 267 (10), 2916-2920CODEN: EJBCAI; ISSN:0014-2956. (Blackwell Science Ltd.)The flexibility of the structure and compressibility of the resp. active site of cytochromes P 450 3A4 (CYP3A4) and BM-3 (CYP102) were studied using absorption spectroscopy in the UV and visual regions. Conformational changes in the overall protein structures of both CYP3A4 and CYP102 due to the effects of temp. and pressure are reversible. However, the enzymes differ in the properties of their active sites. The CYP3A4 enzyme denatures to the inactive P420 form relatively easy, at 3000 bar over half is converted to P420. The compressibility of its active site is lower than that of CYP102 and is greater with the substrate bound, which is in line with the obsd. lack of a stabilizing effect of the substrate on its conformation under pressure. In contrast, CYP102, although having the most compressible active site among the P450s, possesses a structure that does not denature easily to the inactive (P420) form under pressure. In this respect, it resembles the P 450 isolated from acidothermophilic archaebacteria.
- 55Harris, H. A.; Albert, L. M.; Leathurby, Y.; Malamas, M. S.; Mewshaw, R. E.; Miller, C. P.; Kharode, Y. P.; Marzolf, J.; Komm, B. S.; Winneker, R. C.; Frail, D. E.; Henderson, R. A.; Zhu, Y.; Keith, J. C. Evaluation of an Estrogen Receptor-β Agonist in Animal Models of Human Disease. Endocrinology 2003, 144, 4241– 4249, DOI: 10.1210/en.2003-055055https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD3sXns1Srt7o%253D&md5=ed1255fd36790a7b9bc665624e5a4619Evaluation of an estrogen receptor-β agonist in animal models of human diseaseHarris, Heather A.; Albert, Leo M.; Leathurby, Yelena; Malamas, Michael S.; Mewshaw, Richard E.; Miller, Chris P.; Kharode, Yogendra P.; Marzolf, James; Komm, Barry S.; Winneker, Richard C.; Frail, Donald E.; Henderson, Ruth A.; Zhu, Yuan; Keith, James C., Jr.Endocrinology (2003), 144 (10), 4241-4249CODEN: ENDOAO; ISSN:0013-7227. (Endocrine Society)The discovery of a second estrogen receptor (ER), called ERβ, in 1996 sparked intense interest within the scientific community to discover its role in mediating estrogen action. However, despite more than 6 yr of research into the function of this receptor, its physiol. role in mediating estrogen action remains unclear and controversial. We have developed a series of highly selective agonists for ERβ and have characterized their activity in several clin. relevant rodent models of human disease. The activity of one such compd., ERB-041, is reported here. We conclude from these studies that ERβ does not mediate the bone-sparing activity of estrogen on the rat skeleton and that it does not affect ovulation or ovariectomy-induced wt. gain. In addn., these compds. are nonuterotrophic and nonmammotrophic. However, ERB-041 has a dramatic beneficial effect in the HLA-B27 transgenic rat model of inflammatory bowel disease and the Lewis rat adjuvant-induced arthritis model. Daily oral doses as low as 1 mg/kg reverse the chronic diarrhea of HLA-B27 transgenic rats and dramatically improve histol. disease scores in the colon. The same dosing regimen in the therapeutic adjuvant-induced arthritis model reduces joint scores from 12 (maximal inflammation) to 1 over a period of 10 d. Synovitis and Mankin (articular cartilage) histol. scores are also significantly lowered (50-75%). These data suggest that one function of ERβ may be to modulate the immune response, and that ERβ-selective ligands may be therapeutically useful agents to treat chronic intestinal and joint inflammation.
- 56Fulmer, G. R.; Miller, A. J. M.; Sherden, N. H.; Gottlieb, H. E.; Nudelman, A.; Stoltz, B. M.; Bercaw, J. E.; Goldberg, K. I. NMR Chemical Shifts of Trace Impurities: Common Laboratory Solvents, Organics, and Gases in Deuterated Solvents Relevant to the Organometallic Chemist. Organometallics 2010, 29, 2176– 2179, DOI: 10.1021/om100106e56https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXkslKms7Y%253D&md5=53317e05fe3be77fe7de48d8fe1fb8b7NMR Chemical Shifts of Trace Impurities: Common Laboratory Solvents, Organics, and Gases in Deuterated Solvents Relevant to the Organometallic ChemistFulmer, Gregory R.; Miller, Alexander J. M.; Sherden, Nathaniel H.; Gottlieb, Hugo E.; Nudelman, Abraham; Stoltz, Brian M.; Bercaw, John E.; Goldberg, Karen I.Organometallics (2010), 29 (9), 2176-2179CODEN: ORGND7; ISSN:0276-7333. (American Chemical Society)Tables of 1H and 13C NMR chem. shifts have been compiled for common org. compds. often used as reagents or found as products or contaminants in deuterated org. solvents. Building upon the work of Gottlieb, Kotlyar, and Nudelman in the Journal of Org. Chem., signals for common impurities are now reported in addnl. NMR solvents (tetrahydrofuran-d8, toluene-d8, dichloromethane-d2, chlorobenzene-d5, and 2,2,2-trifluoroethanol-d3) which are frequently used in organometallic labs. Chem. shifts for other orgs. which are often used as reagents or internal stds. or are found as products in organometallic chem. are also reported for all the listed solvents.
- 57Thirumamagal, B. T. S.; Zhao, X. B.; Bandyopadhyaya, A. K.; Johnsamuel, J.; Tiwari, R.; Golightly, D. W.; Patel, V.; Jehning, B. T.; Backer, M. V.; Barth, R. F.; Lee, R. J.; Backer, J. M.; Tjarks, W. Receptor-Targeted Liposomal Delivery of Boron-Containing Cholesterol Mimics for Boron Neutron Capture Therapy (BNCT). Bioconjugate Chem. 2006, 17, 1141– 1150, DOI: 10.1021/bc060075d57https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD28XotV2ltbs%253D&md5=4a6a65107118389c48d5ee8233e563f8Receptor-Targeted Liposomal Delivery of Boron-Containing Cholesterol Mimics for Boron Neutron Capture Therapy (BNCT)Thirumamagal, B. T. S.; Zhao, Xiaobin B.; Bandyopadhyaya, Achintya K.; Narayanasamy, Sureshbabu; Johnsamuel, Jayaseharan; Tiwari, Rohit; Golightly, Danold W.; Patel, Vimalkumar; Jehning, Brian T.; Backer, Marina V.; Barth, Rolf F.; Lee, Robert J.; Backer, Joseph M.; Tjarks, WernerBioconjugate Chemistry (2006), 17 (5), 1141-1150CODEN: BCCHES; ISSN:1043-1802. (American Chemical Society)Liposomes have been a main focus of tumor-selective boron delivery strategies in boron neutron capture therapy (BNCT), a binary method for the treatment of cancer that is based on the nuclear reaction between boron atoms and low-energy thermal neutrons. Three novel carboranyl cholesterol derivs. were prepd. as lipid bilayer components for the construction of nontargeted and receptor-targeted boronated liposomes for BNCT. A major structural feature of these novel boronated cholesterol mimics is the replacement of the B and the C ring of cholesterol with a carborane cluster. Computational analyses indicated that all three boronated compds. have structural features and physicochem. properties that are very similar to those of cholesterol. One of the synthesized boronated cholesterol mimics was stably incorporated into non-, folate receptor (FR)-, and vascular endothelial growth factor receptor-2 (VEGFR-2)-targeted liposomes. No major differences were found in appearance, size distribution, and lamellarity between conventional dipalmitoylphosphatidylcholine (DPPC)/cholesterol liposomes, nontargeted, and FR-targeted liposomal formulations of this carboranyl cholesterol deriv. FR-targeted boronated liposomes were taken up extensively in FR overexpressing KB cells in vitro, and the uptake was effectively blocked in the presence of free folate. In contrast, a boronated cholesterol mimic incorporated into nontargeted liposomes showed significantly lower cellular uptake. There was no apparent in vitro cytotoxicity in FR overexpressing KB cells and VEGFR-2 overexpressing 293/KDR cells when these were incubated with boronated FR- and (VEGFR-2)-targeted liposomes, resp., although the former accumulated extensively in KB cells and the latter effectively interacted with VEGFR-2 by causing autophosphorylation and protecting 293/KDR cells from SLT (Shiga-like toxin)-VEGF cytotoxicity.
- 58Fujii, S.; Yamada, A.; Nakano, E.; Takeuchi, Y.; Mori, S.; Masuno, H.; Kagechika, H. Design and Synthesis of Nonsteroidal Progesterone Receptor Antagonists Based on C,C′-Diphenylcarborane Scaffold as a Hydrophobic Pharmacophore. Eur. J. Med. Chem. 2014, 84, 264– 277, DOI: 10.1016/j.ejmech.2014.07.03458https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2cXht1Grs7bF&md5=5b524301e7d8a908c6848a54652d2dc5Design and synthesis of nonsteroidal progesterone receptor antagonists based on C,C'-diphenylcarborane scaffold as a hydrophobic pharmacophoreFujii, Shinya; Yamada, Ayumi; Nakano, Eiichi; Takeuchi, Yuki; Mori, Shuichi; Masuno, Hiroyuki; Kagechika, HiroyukiEuropean Journal of Medicinal Chemistry (2014), 84 (), 264-277CODEN: EJMCA5; ISSN:0223-5234. (Elsevier Masson SAS)The progesterone receptor (PR) plays important roles in multiple physiol. processes, including female reprodn. Here, we report the synthesis of nonsteroidal PR antagonists contg. a boron cluster as the hydrophobic core. We found that 1,7-diphenyl-meta-carborane was the preferred substructure among the three carborane isomers. One compd. was a potent PR antagonist (IC50: 29 nM). Another compd. also exhibited potent activity (IC50: 93 nM), and did not bind to androgen receptor, glucocorticoid receptor or mineralocorticoid receptor. These compds. may be useful for investigating potential clin. applications of PR modulators.
- 59Sedlak, D.; Paguio, A.; Bartunek, P. Two Panels of Steroid Receptor Luciferase Reporter Cell Lines for Compound Profiling. Comb. Chem. High Throughput Screening 2011, 14, 248– 266, DOI: 10.2174/13862071179522244659https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXnslagtL8%253D&md5=0a021e26cec2615f445f7b3d18625c39Two panels of steroid receptor luciferase reporter cell lines for compound profilingSedlak, David; Paguio, Aileen; Bartunek, PetrCombinatorial Chemistry & High Throughput Screening (2011), 14 (4), 248-266CODEN: CCHSFU; ISSN:1386-2073. (Bentham Science Publishers Ltd.)Steroid hormone receptors represent a major target in drug discovery. As ligand inducible transcription factors, their activity can be modulated by small lipophilic mols. Here we describe two panels of potent and selective luciferase reporter cell lines based on cells with low endogenous steroid receptor activity (U2OS). The panels contain reporter cell lines for estrogen receptors α and β, androgen, glucocorticoid, mineralocorticoid, and progesterone receptors. In the first panel, the activation of either synthetic, steroid response elements contg. promoter or viral promoter is mediated by full-length steroid receptors. The second panel is based on the expression of the chimeric receptor, which was created by the replacement of the N-terminal part of the mol. by Gal4 DBD and that binds to multiple UAS sites in the reporter promoter. Both panels were extensively characterized by profiling 28 ligands in dose response manner in agonist and antagonist mode. We have analyzed and compared the responses to tested ligands from both panels and concluded that in general both systems generated similar qual. response in terms of potency, efficacy, partial agonism/antagonism, mixed agonistic/antagonistic profiles and the rank of potencies was well conserved between both panels. However, we have also identified some artifacts introduced by the Gal4/LBD reporter assays in contrast to their full-length receptor reporter counterparts. Keeping in mind the advantages and drawbacks of each reporter format, these cell lines represent powerful and selective tools for profiling large compd. libraries (HTS) and for detailed study of mechanisms by which compds. exert their biol. effects.
- 60Yung-Chi, C.; Prusoff, W. H. Relationship between the Inhibition Constant (KI) and the Concentration of Inhibitor Which Causes 50 per Cent Inhibition (I50) of an Enzymatic Reaction. Biochem. Pharmacol. 1973, 22, 3099– 3108, DOI: 10.1016/0006-2952(73)90196-2There is no corresponding record for this reference.
- 61Chang, T. K. H.; Waxman, D. J. Enzymatic Analysis of CDNA-Expressed Human CYP1A1, CYP1A2, and CYP1B1 with 7-Ethoxyresorufin as Substrate. In Cytochrome P450 Protocols; Phillips, I. R.; Shephard, E. A., Eds.; Methods in Molecular Biology; Humana Press: Totowa, NJ, 2006; pp 85– 90.There is no corresponding record for this reference.
- 62Souček, P. Novel Sensitive High-Performance Liquid Chromatographic Method for Assay of Coumarin 7-Hydroxylation. J. Chromatogr. B: Biomed. Sci. Appl. 1999, 734, 23– 29, DOI: 10.1016/S0378-4347(99)00325-462https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK1MXns1Ghtbw%253D&md5=313b3ef1e8521fa185c493775b679005Novel sensitive high-performance liquid chromatographic method for assay of coumarin 7-hydroxylationSoucek, PavelJournal of Chromatography B: Biomedical Sciences and Applications (1999), 734 (1), 23-29CODEN: JCBBEP; ISSN:0378-4347. (Elsevier Science B.V.)In this paper, a novel HPLC-based method with fluorometric detection of coumarin 7-hydroxylase is presented. The described method provides a time-effective, more sensitive and specific alternative to the previously used spectrofluorometric assay. Using the developed method, metab. of coumarin in 11 samples of human liver microsomes was evaluated and 1790±690 pmol/min/nmol cytochrome P 450 (CYP) activity was found. Kinetic parameters and linearity of coumarin 7-hydroxylation were studied in a reconstituted system consisting of recombinant CYP2A6 expressed in Escherichia coli, rat NADPH-CYP reductase and usual components. It was found that a 3.5 to 30 min time of incubation is suitable for estn. of coumarin 7-hydroxylase activity. Obsd. Km and Vmax values in the CYP2A6 reconstituted system were 1.48±0.37 μM and 3360±180 pmol product/min/nmol CYP, resp.
- 63Donato, M. T.; Jiménez, N.; Castell, J. V.; Gómez-Lechón, M. J. Fluorescence-Based Assays for Screening Nine Cytochrome P450 (P450) Activities in Intact Cells Expressing Individual Human P450 Enzymes. Drug Metab. Dispos. 2004, 32, 699– 706, DOI: 10.1124/dmd.32.7.69963https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXltlWqt7k%253D&md5=10a4fda9a830c33436a8b2373e3af799Fluorescence-based assays for screening nine cytochrome P450 (P450) activities in intact cells expressing individual human P450 enzymesDonato, M. Teresa; Jimenez, Nuria; Castell, Jose V.; Gomez-Lechon, M. JoseDrug Metabolism and Disposition (2004), 32 (7), 699-706CODEN: DMDSAI; ISSN:0090-9556. (American Society for Pharmacology and Experimental Therapeutics)Here, the authors describe a battery of fluorescence assays for rapid measurement in intact cells of the activity of 9 cytochromes P 450 (P 450) isoforms involved in drug metab. The assays are based on the direct incubation of monolayers of cells expressing individual P 450 enzymes with a fluorogenic substrate followed by fluorometric quantification of the product formed and released into incubation medium. For each individual P 450 activity, different fluorescent probes were examd., and the one showing the best properties (highest metabolic rates, lowest background fluorescence) was selected: 3-cyano-7-ethoxycoumarin for CYP1A2 and CYP2C19, coumarin for CYP2A6, 7-ethoxy-4-trifluoromethylcoumarin for CYP2B6, dibenzylfluorescein for CYP2C8, 7-methoxy-4-trifluoromethylcoumarin (MFC) for CYP2C9 and CYP2E1, 3-[2-(N,N-diethyl-N-methylammonium)ethyl]-7-methoxy-4-methylcoumarin for CYP2D6, and 7-benzyloxy-4-trifluoromethylcoumarin for CYP3A4. The fluorescence-based assays were highly sensitive and allowed the simultaneous measurement of a large no. of samples using plate readers, thus enhancing sample throughput. The major advantages over high-throughput assays in subcellular fractions were that, as living cells were used, manual handling and enzyme damage were minimized, the endoplasmic reticulum of the cells remained intact, exogenous cofactors or NADPH-regenerating systems were not required, and transport processes were maintained. These assays could be applied to preliminary screening of inhibitory effects of new drugs on individual P 450 enzymes. After comparison of the results obtained using the fluorescent probes in intact P 450-expressing cells and those obtained using the HPLC-based selective assays in the same cells, in primary human hepatocytes, or in human liver microsomes, a fairly good agreement was found.
- 64Crespi, C. L.; Chang, T. K. H.; Waxman, D. J. High-Performance Liquid Chromatography Analysis of CYP2C8-Catalyzed Paclitaxel 6α-Hydroxylation. In Cytochrome P450 Protocols; Phillips, I. R.; Shephard, E. A., Eds.; Methods in Molecular Biology; Humana Press: Totowa, NJ, 2006; pp 103– 107.There is no corresponding record for this reference.
- 65Crespi, C. L.; Chang, T. K. H.; Waxman, D. J. Determination of CYP2C9-Catalyzed Diclofenac 4′-Hydroxylation by High-Performance Liquid Chromatography. In Cytochrome P450 Protocols; Phillips, I. R.; Shephard, E. A., Eds.; Methods in Molecular Biology; Humana Press: Totowa, NJ, 2006; pp 109– 113.There is no corresponding record for this reference.
- 66Crespi, C. L.; Chang, T. K. H.; Waxman, D. J. CYP2C19-Mediated (S)-Mephenytoin 4′-Hydroxylation Assayed by High-Performance Liquid Chromatography With Radiometric Detection. In Cytochrome P450 Protocols; Phillips, I. R.; Shephard, E. A., Eds.; Methods in Molecular Biology; Humana Press: Totowa, NJ, 2006; pp 115– 119.There is no corresponding record for this reference.
- 67Crespi, C. L.; Chang, T. K. H.; Waxman, D. J. CYP2D6-Dependent Bufuralol 1′-Hydroxylation Assayed by Reverse-Phase Ion-Pair High-Performance Liquid Chromatography With Fluorescence Detection. In Cytochrome P450 Protocols; Phillips, I. R.; Shephard, E. A., Eds.; Methods in Molecular Biology; Humana Press: Totowa, NJ, 2006; pp 121– 125.There is no corresponding record for this reference.
- 68Lucas, D.; Menez, J.-F.; Berthou, F. [12] Chlorzoxazone: An in Vitro and in Vivo Substrate Probe for Liver CYP2E1. In Methods in Enzymology; Johnson, E. F.; Waterman, M. R., Eds.; Cytochrome P450, Part B; Academic Press, 1996; Vol. 272, pp 115– 123.There is no corresponding record for this reference.
- 69Ghosal, A.; Satoh, H.; Thomas, P. E.; Bush, E.; Moore, D. Inhibition and Kinetics of Cytochrome P4503A Activity in Microsomes from Rat, Human, and Cdna-Expressed Human Cytochrome P450. Drug Metab. Dispos. 1996, 24, 940– 94769https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK28XlvV2itrg%253D&md5=fb95979ee683d0166e6159ccd3bee9e2Inhibition and kinetics of cytochrome P4503A activity in microsomes from rat, human, and cDNA-expressed human cytochrome P450Ghosal, Anima; Satoh, Hiroko; Thomas, Paul E.; Bush, Ernest; Moore, DavidDrug Metabolism and Disposition (1996), 24 (9), 940-947CODEN: DMDSAI; ISSN:0090-9556. (Williams & Wilkins)Midazolam (MDZ) is metabolized in human liver microsomes by the cytochrome P 450 (CYP) 3A subfamily to 1'-hydroxy (1'-OH) and 4-hydroxy (4-OH) metabolites. MDZ is metabolized in the rat primarily to 4-OH MDZ, 1'-OH MDZ, and 1',4-dihydroxy (1',4-diOH) MDZ. The kinetics of 4-OH and 1'-OH metabolite formation were detd. using hepatic microsomes from control, Ro 23-7637 and dexamethasone-treated male rats. KM values for the major metabolite, 4-OH MDZ, were 24.5, 43.1, and 32.8 μM, and the corresponding Vmax values were 5.9, 28.9, and 13 nmol/mg/min for the control, DEX, and Ro 23-7637-treated animals, resp. KM values for 1'-hydroxylation of MDZ (the major metabolite) after incubation with human liver microsomes from three individuals were 5.57, 2.50, and 3.56 μM, and the corresponding Vmax values were 4.38, 0.49, and 0.19 nmol/mg/min, resp. In parallel studies using cDNA-expressed human CYP3A4 microsomes, the KM for 1'-OH formation was 1.56 μM, and the corresponding Vmax was 0.16 nmol/mg/min. MDZ was not metabolized by cDNA-expressed human CYP2D6, CYP2E1, or CYP1A2, thus confirming that these isoforms were not responsible for its biotransformation. The formation of 1',4-diOH metabolite in rat and 1'-OH formation in cDNA-expressed human CYP3A4 microsomes showed a decrease in velocity at high substrate concns. Inhibition studies showed that MDZ hydroxylation was strongly inhibited by ketoconazole and Ro 23-7637 in rat, human, and cDNA-expressed human CYP3A4 microsomes. α-Naphthoflavone stimulated 1'-OH metabolite formation in human and cDNA-expressed human CYP3A4 microsomes at low concn. (10 μM). Naringenin, a flavonoid present in grapefruit juice, also inhibited MDZ metab. in human liver microsomes. Immunoinhibition studies revealed that polyclonal anti-rat CYP3A2 antibody inhibited MDZ metab. 80-90% in rat, human, and cDNA-expressed human CYP3A4 microsomes, thus suggesting that members of the CYP3A4 subfamily were involved in the metab.
- 70Kronbach, T.; Mathys, D.; Umeno, M.; Gonzalez, F. J.; Meyer, U. A. Oxidation of Midazolam and Triazolam by Human Liver Cytochrome P450IIIA4. Mol. Pharmacol. 1989, 36, 89– 9670https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaL1MXlt1Kgu7g%253D&md5=6ecf9fc3682977542e46d9a32be16f21Oxidation of midazolam and triazolam by human liver cytochrome P450IIIA4Kronbach, Thomas; Mathys, Daniel; Umeno, Morio; Gonzalez, Frank J.; Meyer, Urs A.Molecular Pharmacology (1989), 36 (1), 89-96CODEN: MOPMA3; ISSN:0026-895X.The metab. of midazolam and triazolam to their 1'-hydroxy and 4-hydroxy metabolites was studied in microsomes of human livers. The formation of both metabolites was inhibited >90% by an antiserum directed against a pregnenolone 16α-carbonitrile-inducible cytochrome P 450 (P450PCN1) of rat liver. Moreover, midazolam hydroxylase activity was immunopptd. from solubilized human microsomes with polyclonal antibodies against rat P450PCN1 and the closely related human isoenzyme P450NF. A close correlation was obsd. between the amt. of protein detected in immunoblots with these antibodies and the midazolam or triazolam or triazolam hydroxylase activity. The formation of both metabolites of midazolam was inhibited by triacetyloleandomycin, a known inhibitor of cytochromes P 450 of the IIIA family. Direct evidence that P450IIIA4 catalyzes the metab. of midazolam was provided through the use of cDNA-directed expression. Monkey COS cells transfected with human P450PCN1 cDNA were able to catalyze both the 1'- and the 4-hydroxylation of midazolam. The metab. of midazolam and triazolam in human liver is apparently predominantly mediated by cytochrome P450IIIA4. Two of 15 human livers expressed a 2nd immunoreactive microsomal protein of higher apparent mol. wt. and were more active in midazolam 1'-hydroxylation. The data also provide evidence that the marked interindividual variation in the response to these widely used benzodiazepine drugs is due to variable hepatic metab.
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Activity profiles of library compounds in cell-based reporter assays for nuclear receptors; methods and experimental conditions for cytochrome P450 enzyme assays; and NMR and MS spectra and HPLC chromatograms (PDF)
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