Hydrochloric Acid → Phosphate

HRID 2303921ノート

反応式

HRID 2303921 の反応式
[P:1]([O-:5])([O-:4])([O-:3])=[O:2].[C:6]([OH:27])(=[O:26])[CH2:7][CH2:8][CH2:9][CH2:10][CH2:11][CH2:12][CH2:13][CH2:14][CH2:15][CH2:16][CH2:17][CH2:18][CH2:19][CH2:20][CH2:21][CH2:22][CH2:23][CH2:24][CH3:25].Cl.C([O:34]C(C1C=CC(O)=CC=1)=O)CC(C)C>CS(C)=O.O.[Cl-].[Ca+2].[Cl-].C([C@H](N)C(O)=O)CC(N[C@H](C(NCC(O)=O)=O)CS)=O.C(OCC)(=O)C>[P:1]([O-:5])([O-:4])([O-:3])=[O:2].[CH3:25][CH2:24][CH2:23][CH2:22][CH2:21][CH:20]=[CH:19][CH2:18][CH:17]=[CH:16][CH2:15][CH:14]=[CH:13][CH:12]=[CH:11][CH:10]([OH:34])[CH2:9][CH2:8][CH2:7][C:6]([OH:27])=[O:26]

参加物

11

反応物 4

HCID 313Hydrochloric AcidHCID 1061PhosphateHCID 10467Eicosanoic AcidHCID 81009Isoamyl 4-hydroxybenzoate

生成物 2

HCID 1061PhosphateHCID 17715-Hydroxyicosa-6,8,11,14-tetraenoic acid

試薬・触媒・溶媒 5

HCID 679Dimethyl Sulfoxide溶媒HCID 962Water溶媒HCID 8857Ethyl Acetate溶媒HCID 124886Glutathione溶媒HCID 5284359Calcium Chloride溶媒

反応条件

温度
25 °C

実験手順

Into 5 μl of a solution of the test compound in dimethylsulfoxide, 750 μl of a 1/15M phosphate buffer (pH 7.4) which was prepared by dissolving a wrapper of phosphate buffer powder (Yatron) in 10 ml of water, 40 μl of a 30 mM aqueous calcium chloride solution, 50 μl of a 20 mM aqueous GSH (glutathione, reduced form) solution, 50 μl of a 40 mM aqueous ATP (adenosine-5-triphosphate) solution and 100 μl of guinea pig polymorphonuclear leukocyte enzyme were added, followed by stirring at 25° C. for 5 minutes. To initiate the reaction, 5 μl of arachidic acid (10 mg/ml ethanol) were added to said mixture and the mixture was stirred at 25° C. for 5 minutes. 50 μl of 2N hydrochloric acid were added to the reaction mixture to terminate the reaction. 2.0 ml of ethyl acetate containing an internal standard substance (internal standard substance: 2 μg/ml of isoamyl paraben) were then added to the reaction mixture. The resulting mixture was extracted for 1 minute by a mixer, followed by centrifugal separation at 3000 rpm for 5 minutes. The ethyl acetate layer was separated, evaporated to dryness under reduced pressure and then dissolved in 200 μl of acetonitrile. By high-performance liquid chromatography, the amount of 5-HETE formed was determined. From the determination of 5-HETE of the control without the test compounds and that of the test compounds, an inhibition rate (%) was found and the 50% inhibition concentration of the test compounds against 5-lipoxygenase was determined. The results are shown in Table 12.

後処理

  1. additionwere added
  2. additionwere added
  3. stirringthe mixture was stirred at 25° C. for 5 minutes
  4. customto terminate
  5. customthe reaction
  6. extractionThe resulting mixture was extracted for 1 minute by a mixer
  7. customfollowed by centrifugal separation at 3000 rpm for 5 minutes
  8. customThe ethyl acetate layer was separated
  9. customevaporated to dryness under reduced pressure
  10. dissolutiondissolved in 200 μl of acetonitrile

出典と根拠

ORDレコード
ord-4b1387a740974c09867e0488e5c96dd6
出典データセット
uspto-grants-2002_08
特許番号
US06432993B1

反応ノート

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