Adenosine Monophosphate → Camp

HRID 1288784

反応式

HRID 1288784 の反応式
[CH:1]1[N:6]=[C:5]([NH2:7])[C:4]2[N:8]=[CH:9][N:10]([C@@H:11]3[O:15][C@H:14]([CH2:16][O:17][P:18]([OH:21])([OH:20])=[O:19])[C@@H:13](O)[C@H:12]3[OH:23])[C:3]=2[N:2]=1.C1C(N)=CC=C(SC#N)C=1>>[CH:1]1[N:6]=[C:5]([NH2:7])[C:4]2[N:8]=[CH:9][N:10]([C@@H:11]3[O:15][C@@H:14]4[CH2:16][O:17][P:18]([OH:20])([O:21][C@H:13]4[C@H:12]3[OH:23])=[O:19])[C:3]=2[N:2]=1

参加物

3

反応物 2

HCID 6083Adenosine MonophosphateHCID 76336[(4-Aminophenyl)sulfanyl]formonitrile

生成物 1

HCID 6076Camp

実験手順

The adenylate cyclase activity induced by each of Analogs I-V was also measured in SaOS-2 B10 cells as described previously (Rodan et al. 1983 J. Clin. Invest. 72;1511; Goldman et al. 1988 Endocrinology 123, 1468). Confluent SaOS-2 B10 cells in 24 wells plates were incubated 0.5 μCi[3H]adenine (26.9 Ci/mmol, New England Nuclear, Boston, Mass.) in fresh medium at 37° C. for 2 hrs, and washed twice with Hank's solution. The cells were treated with 1 mM IBMX [isobutylmethylxanthine, Sigma, St. Louis, Mo.] in fresh medium for 15 min, and a tested PTH analog was added to the medium to incubate for 5 min. The reaction was stopped by the addition of 1.2M TCA followed by sample neutralization with 4N KOH. Cyclic AMP was isolated by the two-column chromatographic method (see Salomon et al. 1974 Anal. Biochem. 58;541). The radioactivity was counted in a scintillation counter (Liquid scintillation counter 2200CA, PACKARD, Downers Grove, Ill.). The EC50 's (half maximal stimulation of adenylate cyclase) were calculated for the five PTH analogs and are shown below:

後処理

  1. customat 37° C.
  2. customfor 2 hrs
  3. washwashed twice with Hank's solution
  4. additionThe cells were treated with 1 mM IBMX [isobutylmethylxanthine, Sigma, St. Louis, Mo.] in fresh medium for 15 min
  5. additiona tested PTH analog was added to the medium
  6. additionThe reaction was stopped by the addition of 1.2M TCA

出典と根拠

ORDレコード
ord-dc4bbec37752466daff392b683623f9c
出典データセット
uspto-grants-1998_05
特許番号
US05747456

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