反応式
C1N(CCO)CCN(CCS(O)(=O)=O)C1.C(N(CC(O)=O)CC(O)=O)COCCOCCN(CC(O)=O)CC(O)=O.[Mg+2].[Cl-].[Cl-].[P:45]([O:57][CH2:58][C@H:59]1[O:63][C@@H:62]([N:64]2[C:73]3[N:72]=[CH:71][N:70]=[C:68]([NH2:69])[C:67]=3[N:66]=[CH:65]2)[C@H:61]([OH:74])[C@@H:60]1[OH:75])([O:48][P:49]([O:52][P:53]([OH:56])([OH:55])=[O:54])([OH:51])=[O:50])(=[O:47])[OH:46].[CH3:76][CH2:77][CH2:78][CH2:79][CH2:80][CH2:81][CH2:82][C:83]([O:85][CH2:86][CH:87]([O:90][C:91]([CH2:93][CH2:94][CH2:95][CH2:96][CH2:97][CH2:98][CH3:99])=[O:92])[CH2:88][OH:89])=[O:84].[CH2:100]([OH:128])[CH:101]([OH:127])[CH2:102][O:103][P:104]([O:107][C@H:108]1[C@H:113]([OH:114])[C@@H:112]([O:115][P:116]([OH:119])([OH:118])=[O:117])[C@H:111]([O:120][P:121]([OH:124])([OH:123])=[O:122])[C@@H:110]([OH:125])[C@H:109]1[OH:126])([OH:106])=[O:105]>>[P:45]([O:57][CH2:58][C@H:59]1[O:63][C@@H:62]([N:64]2[C:73]3[N:72]=[CH:71][N:70]=[C:68]([NH2:69])[C:67]=3[N:66]=[CH:65]2)[C@H:61]([OH:74])[C@@H:60]1[OH:75])([O:48][P:49]([O:52][P:53]([OH:55])([OH:56])=[O:54])([OH:51])=[O:50])(=[O:46])[OH:47].[CH3:76][CH2:77][CH2:78][CH2:79][CH2:80][CH2:81][CH2:82][C:83]([O:85][CH2:86][CH:87]([O:90][C:91]([CH2:93][CH2:94][CH2:95][CH2:96][CH2:97][CH2:98][CH3:99])=[O:92])[CH2:88][OH:89])=[O:84].[CH2:100]([OH:128])[CH:101]([OH:127])[CH2:102][O:103][P:104]([O:107][C@H:108]1[C@H:113]([OH:114])[C@@H:112]([O:115][P:116]([OH:118])([OH:119])=[O:117])[C@H:111]([O:120][P:121]([OH:124])([OH:123])=[O:122])[C@@H:110]([OH:125])[C@H:109]1[OH:126])([OH:106])=[O:105]参加物
実験手順
Promega ADP-Glo Max assay kit (Cat. No. V7002) was utilized to determine IC50 values for α, β, δ and γ isoforms of human Class I PI3 kinases (Millipore). Samples of kinase (20 nM α or δ, 40 nM β or γ isoform) were incubated with compound for 15 minutes at room temperature in reaction buffer (15 mM HEPES pH 7.4, 20 mM NaCl, 1 mM EGTA, 0.02% Tween 20, 10 mM MgCl2, 0.2 mg/mL bovine-γ-globulins) followed by addition of ATP/diC8-PtdInsP mixture to give final concentrations of 3 mM ATP and 500 uM diC8-PtdInsP. Reactions were incubated at room temperature for 2 hours followed by addition of 25 uL of stop solution. After a 40-minute incubation at room temperature, 50 uL of Promega detection mix was added followed by incubation for 1 hour at room temperature. Plates were then read on Envision plate reader in luminescence mode. Data was converted to % inhibition using the following equation below:
出典と根拠
- ORDレコード
- ord-ae5a39c10eb3435a88f7a8b1fb8715e2
- 出典データセット
- uspto-grants-2015_01
- 特許番号
- US08940742B2
関連ノート
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