Neca → Adenosine

HRID 22066

反応式

HRID 22066 の反応式
CCN[C:4]([C@H:6]1[O:10][C@@H:9]([N:11]2[C:15]3[N:16]=[CH:17][N:18]=[C:19]([NH2:20])[C:14]=3[N:13]=[CH:12]2)[C@H:8]([OH:21])[C@@H:7]1[OH:22])=[O:5].C1([C@@]2(N3C4N=CN=C(N)C=4N=C3)O[C@H](CO)[C@@H](O)[C@H]2O)CCCCC1.[Mg+2].[Cl-].[Cl-]>C1(C(NC2N=C(N)C3N=CN(C=3N=2)[C@@H]2O[C@H](CO)[C@@H](O)[C@H]2O)(C)C)C=CC=CC=1>[C@@H:9]1([N:11]2[C:15]3[N:16]=[CH:17][N:18]=[C:19]([NH2:20])[C:14]=3[N:13]=[CH:12]2)[O:10][C@H:6]([CH2:4][OH:5])[C@@H:7]([OH:22])[C@H:8]1[OH:21]

参加物

5

反応物 3

HCID 448222NecaHCID 5360315Magnesium ChlorideHCID 66601354Cyclohexyladenosine

生成物 1

HCID 60961Adenosine

試薬・触媒・溶媒 1

HCID 54295855(R)-2-[(Phenylisopropyl)Amino]Adenosine溶媒

実験手順

Incubation tubes, in triplicate, receive 100 μl of [3H]NECA (94 nM in the assay), 100 μl of 1 μM cyclohexyladenosine (CHA), 100 μl of 100 mM MgCl2, 100 μl of 1 IU/ml adenosine deaminase, 100 μl of test compounds at various concentrations over the range of 10-10M to 10-4M diluted with assay buffer (50 mM Tris-HCl, pH 7.7) and 0.2 μl of membrane suspension (5 mg wet weight), in a final volume of 1 ml of 50 mM Tris-HCl, pH 7.7. Incubations are carried out at 25° C. for 60 minutes. Each tube is filtered through GF/B glass fiber filters using a vacuum. The filters are rinsed two times with 5 ml of the ice-cold buffer. The membranes on the filters are transferred to scintillation vials to which 8 ml of Omnifluor with 5% Protosol is added. The filters are counted by liquid scintillation spectrometry.

後処理

  1. concentration/ml adenosine deaminase, 100 μl of test compounds at various concentrations over the range of 10-10M to 10-4M
  2. additiondiluted with assay buffer (50 mM Tris-HCl, pH 7.7) and 0.2 μl of membrane suspension (5 mg wet weight)
  3. filtrationEach tube is filtered through GF/B glass fiber filters
  4. washThe filters are rinsed two times with 5 ml of the ice-cold buffer
  5. additionis added

出典と根拠

ORDレコード
ord-20e1f244ba284a2a9325cd7161d23d69
出典データセット
uspto-grants-1995_06
特許番号
US05426101

関連ノート

読み込み中…