Reaktionsgleichung
C(O)[C:2]([NH2:7])([CH2:5][OH:6])[CH2:3][OH:4].C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)C[C:13](O)=[O:14].S[CH2:30]CO.SC[C@H]([C@@H](CS)O)O.CC(C[C@H](NC(C)=O)C(N[C@H](C(N[C@H](C(O)=O)CCCN=C(N)N)=O)CC(C)C)=O)C.[C:72](N)(=O)[C:73]1[CH:78]=[CH:77][CH:76]=[CH:75][CH:74]=1.[CH:81]1[CH:86]=[CH:85][C:84]([CH2:87]S(F)(=O)=O)=[CH:83][CH:82]=1.[CH2:92]1[S:98][C:96](=[O:97])[NH:95][C:93]1=[O:94]>OCC(CO)O>[CH3:30][C:5]1[O:6][C:72]([C:73]2[CH:78]=[CH:77][CH:76]=[CH:75][CH:74]=2)=[N:7][C:2]=1[CH:3]([OH:4])[CH2:13][O:14][C:81]1[CH:82]=[CH:83][C:84]([CH2:87][CH:92]2[S:98][C:96](=[O:97])[NH:95][C:93]2=[O:94])=[CH:85][CH:86]=1Komponenten
Edukt 8
Produkt 1
Reagenzien, Katalysatoren, Lösungsmittel 1
Versuchsvorschrift
For each assay, an aliquot of receptor (1:1000-1:3000 dilution) was incubated in TEGM (10 mM Tris, pH 7.2, 1 mM EDTA, 10% glycerol, 7 μl/100 ml β-mercaptoethanol, 10 mM Na molybdate, 1 mM dithiothreitol, 5 μg/ml aprotinin, 2 μg/ml leupeptin, 2 μg/ml benzamide and 0.5 mM PMSF) containing 5-10% COS-1 cell cytoplasmic lysate and 10 nM labeled thiazolidinedione ([3H2 ]AD-5075, 21 Ci/mmole), ± test compound compound, [3H2 ]Example 11, 17 Ci/mmole), ± test compound, respectively. Assays were incubated for ~16 h at 4 ° C. in a final volume of 300 μl. Unbound ligand was removed by addition of 200 μl dextran/gelatin-coated charcoal, on ice, for ~10 minutes. After centrifugation at 3000 rpm for 10 min at 4° C., 200 μl of the supernatant fraction was counted in a liquid scintillation counter. In this assay the KD for AD-5075 and Example 11 is 1 nM, respectively.
Aufarbeitung
- additioncontaining 5-10% COS-1 cell cytoplasmic lysate and 10 nM
- waitAssays were incubated for ~16 h at 4 ° C.
- customUnbound ligand was removed by addition of 200 μl dextran/gelatin-coated charcoal, on ice, for ~10 minutes
Quellen und Nachweise
- ORD-Eintrag
- ord-02cc420f906c4c0687d2f648116f01e1
- Quelldatensatz
- uspto-grants-2000_02
- Patentnummer
- US06020382
Verknüpfte Notizen
Wird geladen…