A novel estrogen receptor ligand template

Bioorganic and Medicinal Chemistry Letters

Volumn 13, Issue 11, 2003, Pages 1919-1922

  Sibley, Robert ^a   Hatoum Mokdad, Holia ^a   Schoenleber, Robert ^a   Musza, Laszlo ^a   Stirtan, William ^a   Marrero, Diana ^a   Carley, William ^a   Xiao, Hong ^a   Dumas, Jacques ^a  

^a BAYER CORPORATION   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ALCOHOL DERIVATIVE; BICYCLO[3.3.1]NONENE; ESTROGEN RECEPTOR ALPHA; ESTROGEN RECEPTOR BETA; RALOXIFENE; UNCLASSIFIED DRUG;

ARTICLE; DRUG POTENTIATION; DRUG SELECTIVITY; LIGAND BINDING; RECEPTOR BINDING; STEREOCHEMISTRY;

EID: 0038056114     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0960-894X(03)00307-X     Document Type: Article

References (18)

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4. Smith R.A., Chen J., Mader M.M., Muegge I., Moehler U., Katti S., Marrero D., Stirtan W.G., Weaver D.R., Xiao H., Carley W. Bioorg. Med. Chem. Lett. 12:2002;2875. For a recent review on SERM templates, see: Fink B.E., Mortensen D.S., Stauffer S.R., Aron Z.D., Katzenellenbogen J.A. Chem. Biol. 6:1999;205.
5. Smith R.A., Chen J., Mader M.M., Muegge I., Moehler U., Katti S., Marrero D., Stirtan W.G., Weaver D.R., Xiao H., Carley W. Bioorg. Med. Chem. Lett. 12:2002;2875. For a recent review on SERM templates, see: Fink B.E., Mortensen D.S., Stauffer S.R., Aron Z.D., Katzenellenbogen J.A. Chem. Biol. 6:1999;205.
6. 1H NMR analysis of bicyclic ether 2 shows that this sample consists of a 9:1 mixture of two isomers. Stereochemical considerations suggest that the major isomer is also the active ingredient.
7. 6] estradiol (4.8 nM). Antimouse antibody SPA binding beads were then added to each well (0.25 mg/well), the reaction mixture was equilibrated (3 h), the plate centrifuged (2000 rpm, 10 min), and counted in a Microbeta scintillation counter (Wallac, Inc.). Raloxifene 1 was used as a positive control, unlabeled estradiol 4 was used as a non-specific binding control.
8. Crenshaw R.R., Luke G.M., Jenks T.A., Partyka R.A., Bialy G., Bierwagen M.E. J. Med. Chem. 16:1973;813.
9. When using benzyl or methyl acrylate as the dienophile, the ratio of the desired regioisomer 6 to the undesired was 4:1 in each case.
10. Rappoport Z., Gal A. J. Chem. Soc., Perkin Trans. 2. 1973;301.
11. 3SnCl, -78 °C to rt).
12. 13C (ppm) 1 1.88 m 48.6 2 2.57 m 51.8 3a 1.83 m 33.3 3b 1.59 m 4a 1.97 m 35.3 4b 1.25 m 5 43.7 6a 1.94 m 22.0 6b 1.86 m 7 5.62 m 126.1 8 5.36 m 128.1 9a 1.68 m 29.0 9b 1.33 m 10 133.8 11 7.00 d (J=8.5 Hz) 127.9 12 6.66 d (J=8.5 Hz) 115.0 13 155.5 14a 3.27 dd (J=5.5 and 10.5 Hz) 65.6 14b 3.20 dd (J=5.5 and 10.5 Hz) 13 OH 9.13 s 14 OH 4.56 t (J=5.5 Hz) Elucidation of relative stereochemistry was based on NOE data in comparison with molecular models. Molecular mechanics calculations were performed on a Silicon Graphics Indy workstation using Tripos force fields and the Gasteiger and Hückel method for charges as implemented in SYBYL version 6.6 (Tripos, St. Louis, MO, USA). The basis for the stereochemical assignment of bicyclic analogue 3 is the observation of cross-peaks between the olefinic proton H8 and the aromatic proton H11 in the ROESY spectrum, indicating spatial proximity in agreement with distance calculations. The presence of a strong cross-peak between protons H2 and H9a provides further evidence for the assigned stereochemistry.
13. 50 values showed an error of ≤15%.
14. While our data indicate that raloxifene is approximately 70-fold selective for ERα over ERβ, the number has historically been 10- to 20-fold: see: Miller C.P. Curr. Pharm. Des. 8:2002;2089 Miller C.P., Komm B.S. Ann. Rep. Med. Chem. 36:2001;149, and. references cited therein for information on the binding of raloxifene and other SERMs to ERα and ERβ
15. While our data indicate that raloxifene is approximately 70-fold selective for ERα over ERβ, the number has historically been 10- to 20-fold: see: Miller C.P. Curr. Pharm. Des. 8:2002;2089 Miller C.P., Komm B.S. Ann. Rep. Med. Chem. 36:2001;149, and. references cited therein for information on the binding of raloxifene and other SERMs to ERα and ERβ
16. MCF-7 cells were obtained from the ATCC and grown in Growth Medium (GM, see below). The cells are grown to 80% confluency and trypsinized and washed to remove phenol red, then plated at 10,000 cells/well in Starve Medium (SM, see below). After 2 days, the medium is removed from the cells and either fresh SM or SM plus test compound is added (100 mL total volume). The next day the bDNA assay is started by adding Capture Hybridization Buffer to each capture well of the bDNA plate (all solutions supplied with QuantiGene kit). Subsequently, medium is removed from the 96-well plate containing cells and then the solution of lysis buffer and oligomers (designed by ProbeDesigner 1.0 for the gene of interest) is added to the 96-well plate (100 μL/well). The cell and lysis/oligomer mixture is incubated for 15 min at 53°C and then vortexed for 1 min. The lysate is then transferred to the bDNA capture plate and mixed. The plate is then sealed and incubated at 53°C for 16 h. The next day, the plate is cooled for 10 min at room temperature and the Amplifier Reagent mixture is prepared. The wells are then washed twice with 200 μL of Wash Solution A. Amplifier Reagent is added (50 μL/well) and the plate is sealed again and incubated at 53°C for 30 min. The plate is then cooled for 10 min at room temperature (rt) and the Label Probe Reagent is prepared. The plate is washed twice with 200 μL of Wash Solution A and 50 μL/well of Label Probe Reagent is added. The plate is sealed and incubated at 53°C for 15 min. The plate is cooled for 10 min at rt and the Substrate Mixture is prepared. The plate is again washed twice with 200 μL Wash Solution A and then twice with Wash Solution D. The Substrate Mixture is then added (50 μL) and the plate is sealed. The plate is incubated at 37°C for 30 min and then read on the Quantiplex luminometer. Raloxifene is used as a positive control in this assay, and behaves as a full antagonist of estrogen-induced pS2 expression.
17. pS2 is an estrogen-responsive gene identified in breast cancer cell lines. Rio M.C., Chambon P. Cancer Cells. 2:1990;269 Davidson N.E., Bronzert D.A., Chambon P., Gelmann E.P., Lippman M.E. Cancer Res. 46:1986;1904.
18. pS2 is an estrogen-responsive gene identified in breast cancer cell lines. Rio M.C., Chambon P. Cancer Cells. 2:1990;269 Davidson N.E., Bronzert D.A., Chambon P., Gelmann E.P., Lippman M.E. Cancer Res. 46:1986;1904.