Hydrochloric Acid → HCID 178501463, compound

HRID 1660432Notes

Reaction equation

HRID 1660432 reaction equation
[CH3:1][O:2][C:3](=[O:23])[C@H:4]([CH2:16][CH2:17][CH2:18][NH:19][C:20](=[NH:22])[NH2:21])[NH:5][S:6]([C:9]1[CH:15]=[CH:14][C:12]([CH3:13])=[CH:11][CH:10]=1)(=[O:8])=[O:7].P([O-])([O-])([O-])=O.NO.NO.Cl.[OH-].[Na+].[Cl:36]C(Cl)(Cl)C(O)=O.Cl>>[ClH:36].[CH3:1][O:2][C:3](=[O:23])[C@H:4]([CH2:16][CH2:17][CH2:18][NH:19][C:20](=[NH:21])[NH2:22])[NH:5][S:6]([C:9]1[CH:15]=[CH:14][C:12]([CH3:13])=[CH:11][CH:10]=1)(=[O:7])=[O:8]

Participants

8

Reactant 7

HCID 313Hydrochloric AcidHCID 787HydroxylamineHCID 1061PhosphateHCID 6421Trichloroacetic AcidHCID 14798Sodium HydroxideHCID 443297Hydroxyamine hydrochlorideHCID 178501462HCID 178501462, compound

Product 1

HCID 178501463HCID 178501463, compound

Procedure

To 0.5 ml of urinary kallikrein were added 0.4 ml of Nα-tosylarginine methyl ester solution (10 micromoles/0.4 ml of 5% DMSO) and 0.1 ml of phosphate buffer (pH 7.4). The mixture was subjected to incubation at 37° C. for 30 min, and 1.5 ml of hydroxylamine solution (a mixture of equal amounts of 2 M NH2OH hydrochloride and 3.5 M NaOH) was added thereto, after which the mixture was allowed to stand at room temperature for 15 min. Thereto were added 1 ml of 18% trichloroacetic acid solution, 1 ml of 4 N hydrochloric acid and 1 ml of 10% ferric chloride solution, and the resulting mixture was stirred thoroughly and then centrifuged at 3,000 r.p.m. for 10 min. The color developed in the supernatant was measured as absorbance (530 nm) by means of a spectrophotometer. The value obtained corresponds to the amount of the substrate remaining unhydrolyzed with kallikrein, and therefore, the activity of the enzyme corresponds to the difference between the value obtained when no enzyme was used (control) and the value obtained after the enzyme reaction.

Workup

  1. additionwas added
  2. waitto stand at room temperature for 15 min
  3. waitfor 10 min
  4. customThe value obtained corresponds to the amount of the substrate
  5. customthe activity of the enzyme corresponds to the difference between the value obtained when no enzyme
  6. customthe value obtained
  7. customafter the enzyme reaction

Sources & evidence

ORD record
ord-4c073cfc4687407985f0bdc0ca70de3a
Source dataset
uspto-grants-1981_12
Patent number
US04308202

Reaction notes

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