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14
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38949119757
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Dols, P.P.M.A.1
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Hermkens, P.H.H.8
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16
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68349134529
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note
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The PR binding assay was performed according to the manufacturers protocol (PR Competitor Assay Kit, Red-(Invitrogen-Product No. P2962)) with minor amendments. Briefly, 40 nM PR-Ligand Binding Domain, 2 nM Fluormone PL Red and 1 mM DTT were dissolved and mixed in Complete PR RED Buffer supplemented with 2 mM CHAPS. Ten microlitres of the mix were dispensed to each well of Greiner low volume plates, containing compounds at the required concentration. The plates were spun for 1 min at 200 g, covered to protect the reagents from light, and then incubated at room temperature for approximately 2 h. Plates were read on an Acquest using a 530-25 nm excitation and 580-10 nm emission interference filter and a 561 nm Dichroic mirror.
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17
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68349156815
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note
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50 = 9.8, 100% efficacy).
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18
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68349140749
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note
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16. In short, 120 μL of a T47D cell suspension was seeded into a 96-well plate and cells were allowed to attach to the plate overnight. On the next day, the cells were treated with compound and incubated overnight. On the following day, 100 μL of pNPP-SPAP was added and there were allowed to stand in the dark for 2 h. Optical density was then measured at a wavelength of 405 nm on a plate reader.
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19
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0031519350
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Nesterov V.N., Krivokolysko S.G., Dyachenko V.D., Dotsenko V.V., and Litvinov V.P. Russ. Chem. Bull. 46 (1997) 990
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(1997)
Russ. Chem. Bull.
, vol.46
, pp. 990
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Nesterov, V.N.1
Krivokolysko, S.G.2
Dyachenko, V.D.3
Dotsenko, V.V.4
Litvinov, V.P.5
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22
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68349134528
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note
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All tricyclic pyridines were prepared and tested as 1:1 mixtures of two enantiomers unless stated otherwise.
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23
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0031181346
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The co-crystal structure of progesterone in PR (PDB code 1A28) was used as the starting point for the initial docking calculations using the program, Flo+, version 0203. Residues within a 15 Å sphere of progesterone in the binding site were retained. The mcdock algorithm, which relies on a Monte Carlo perturbation/fast search/energy minimization algorithm was used. Only those residues that were known to be flexible from other crystal structures were made flexible during the energy minimization step. Two thousand steps of perturbation were performed and the 25 top-ranked poses were retained. Visual inspection of the interactions made by the ligand within the active site and the relative strain energies of the ligand calculated using Flo+ in each pose were used to determine the best docked pose. Also see:
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The co-crystal structure of progesterone in PR (PDB code 1A28) was used as the starting point for the initial docking calculations using the program, Flo+, version 0203. Residues within a 15 Å sphere of progesterone in the binding site were retained. The mcdock algorithm, which relies on a Monte Carlo perturbation/fast search/energy minimization algorithm was used. Only those residues that were known to be flexible from other crystal structures were made flexible during the energy minimization step. Two thousand steps of perturbation were performed and the 25 top-ranked poses were retained. Visual inspection of the interactions made by the ligand within the active site and the relative strain energies of the ligand calculated using Flo+ in each pose were used to determine the best docked pose. Also see:. McMartin C., and Bohacek R.S. J. Comput. Aided Mol. Des. 11 (1997) 333
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(1997)
J. Comput. Aided Mol. Des.
, vol.11
, pp. 333
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McMartin, C.1
Bohacek, R.S.2
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24
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68349140750
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note
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2, 140 bar, 40 °C, UV @ 280 nm; (b) Stereochemistry of 9 and 10 was assigned by Vibrational Circular Dichroism (VCD) analysis of both enantiomers.
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25
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68349129599
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note
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The crystal of PR LBD bound to compound 9 had a data set collected at 17-ID of the APS, and were solved using the programs hkl2000, ccp4 (molrep), coot, and refmac.
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26
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34250894870
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Madauss K.P., Grygielko E.T., Deng S.-J., Sulpizio A.C., Stanley T.B., Wu C., Short S.A., Thompson S.K., Stewart E.L., Laping N.J., Williams S.P., and Bray J.D. Mol. Endocrinol. 21 (2007) 1066
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(2007)
Mol. Endocrinol.
, vol.21
, pp. 1066
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Madauss, K.P.1
Grygielko, E.T.2
Deng, S.-J.3
Sulpizio, A.C.4
Stanley, T.B.5
Wu, C.6
Short, S.A.7
Thompson, S.K.8
Stewart, E.L.9
Laping, N.J.10
Williams, S.P.11
Bray, J.D.12
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27
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68349130840
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note
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+).
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