NADPH → Cortisol

HRID 1424140

反応式

HRID 1424140 の反応式
C1N=C(N)C2N=CN([C@@H]3O[C@H](COP(OP(OC[C@H]4O[C@@H](N5C=C(C(N)=O)CC=C5)[C@H](O)[C@@H]4O)(O)=O)(O)=O)[C@@H](O)[C@H]3OP(O)(O)=O)C=2N=1.P(OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)(O)(O)=O.[Cl-].[Mg+2].[Cl-].[CH3:68][C@@:69]12[C@H:79]3[C:80]([CH2:82][C@:83]4([CH3:93])[C@@:87]([OH:92])([C:88]([CH2:90][OH:91])=[O:89])[CH2:86][CH2:85][C@H:84]4[C@@H:78]3[CH2:77][CH2:76][C:75]1=[CH:74][C:72](=[O:73])[CH2:71][CH2:70]2)=[O:81].C[C@@]12CC[C@]3(C)C(=CC([C@H]4[C@@]3(C)CC[C@@H]3[C@]4(C)CC[C@H](OC(CCC(O)=O)=O)C3(C)C)=O)[C@@H]1C[C@](C(O)=O)(C)CC2>CS(C)=O.CO.OCC(CO)O>[CH3:68][C@@:69]12[C@H:79]3[C@@H:80]([OH:81])[CH2:82][C@:83]4([CH3:93])[C@@:87]([OH:92])([C:88]([CH2:90][OH:91])=[O:89])[CH2:86][CH2:85][C@H:84]4[C@@H:78]3[CH2:77][CH2:76][C:75]1=[CH:74][C:72](=[O:73])[CH2:71][CH2:70]2

参加物

8

反応物 5

HCID 5884NADPHHCID 222786CortisoneHCID 439958Glucose-6-Phosphate2 回HCID 636403CarbenoxoloneHCID 5360315Magnesium Chloride

生成物 1

HCID 5754Cortisol

試薬・触媒・溶媒 2

HCID 753Glycerol溶媒HCID 22036561Dimethyl sulfoxide methanol溶媒

実験手順

A reaction was performed on a 384-well plate (Greiner Bio One) using a reaction volume of 24 μL, and all the samples were diluted with an assay buffer (50 mM tris buffer, pH 7.4, 10% glycerol). 0.8 mM NADPH, 6 mM glucose 6-phosphate, 0.35 units/mL glucose 6-phosphate dehydrogenase (Sigma Chemical), and 3 mM magnesium chloride and a microsome fraction as an enzyme source were added to the plate, and a solution containing a test compound dissolved in a dimethyl sulfoxide/methanol solution was added at a final concentration of 0.1%. After the addition of the test compound, cortisone at a final concentration of 160 nM was added to start the reaction, and the reaction was performed at room temperature for 3 h. 25 μL of 100 mM carbenoxolone (Sigma Chemical) was added to terminate the reaction, and the amount of cortisol produced was measured with RUBYstar (BMG LABTECH JAPAN Ltd.) using a cortisol prototype kit (Cisbio International) according to the package insert. The IC50 values of the compounds of Examples are shown below.

後処理

  1. additiona solution containing a test compound
  2. additionwas added at a final concentration of 0.1%
  3. additionwas added
  4. customthe reaction
  5. customto terminate
  6. customthe reaction

出典と根拠

ORDレコード
ord-5ff24abd4cfe4af096225f4452a1b8b8
出典データセット
uspto-grants-2015_01
特許番号
US08927536B2

関連ノート

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