Luciferase reporter gene. These cell lines were divided into two steps: rst, an expression cassette for human AR under control of the CMV promoter in any location in the genome of HEK293 Flp-In cell line h integrated. A line AR weight was just increments and second, the FLP system was used to integrate a ARQ 197 luciferase reporter gene under control Of the pGL4 a bo You E1B TATA and four copies of HRE Slp 2 SLP or two mutated HRE in Flp recombination target site. The resulting cell lines were called SelARE and Clare, respectively. Expression of human AR SelARE and Clare was best determined by Western blot analysis CONFIRMS. Known hypersensitivity to androgens such as T, DHT and methyltrienolone was incubated overnight with a dilution series of androgen from 13 hours to 0.3 M. The EC 50 of DHT, T and 0.36 nM R1881 determined for SelARE, 1.81 nM and 0.24 nM. For Claire, the androgen-response was almost identical. Only when androgens such as medroxyprogesterone acetate compounds, Dianabol, 5 饪 it Androstanediol and dihydrotestosterone were added, the reporter system was activated. Ligands other nuclear receptors such as aldosterone, progesterone, Estradiol, vitamin D, dexamethasone and hydroxycorticosterone could stimulate cell lines. For the detection of AR antagonists erm Adjusted, DHT TNF-Alpha Signaling Pathway was coadministered with a submaximal concentration to activate the AR. The success rate for the projection of 1.3% with a false-positive rate of only 0.1%.
This led to the identification of eight 5.6 pyrazinocarbazole dihydro 4H, further structure activity relationship of compound 3 MEL For more information about how molecular bond, chemical derivatives of MEL-3 were purchased and tested. In Mel Mel 3.1 to 3.6, the isopropyl group at position 11 is changed from MEL 3 Ge, W While the aromatic pyrazine 3 is Mel Mel MEL Ver 3.7 to 3.8 Changed. Use of home computing, was the observed structure-activity Ts relationship with a structural point of analysis. MEL 3 docked in the binding of the ligand-binding Dom ne of AR on the binding of DHT to the AR-LBD and the binding of AR-LBD with the Bic basis. To the common-mode bind to these strong links is shown in Figure 2A. The key interaction is the free electron pair of the nitrogen in the 16th M March MEL derivatives formed with Arg 752 of the AR. This interaction with duplication of the interaction of the cyano group by Bic and the carbonyl group of DHT and can be regarded as essential for the binding compounds. Methylation of nitrogen in MEL 3.7 16 prevents the formation of this hydrogen bond from the free electron pair conformation has changed GE And the space is Afatinib now occupied by a proton. W During the MEL 3.8, disables the reduction of pyrazine in a piperazine formation of hydrogen bonds with Arg 572 through the wrong orientation of the lone pair of nitrogen 16th Disruption of this hydrogen bond is thus obtained in the absence of the power of MEL 3.7 and Inaktivit t in the MEL 3.8. The hydrophobic cyclohexyl derivatives MEL 3 overlap well with the hydrophobic trifluoromethyl Bic. In addition, the activity t MEL 3.1, containing a methyl group at position 11, Clare is comparable to the activity T 3 itself MEL