Ammonia → HCID 178455899, compound

HRID 1982319Notes

Reaction equation

HRID 1982319 reaction equation
[NH2:1][C@H:2]([C:10]([NH:12][C@H:13]([C:21]([N:23]1[CH2:90][CH2:89][CH2:88][C@H:24]1[C:25]([NH:27][C@H:28]([C:36]([NH:38][C@H:39]([C:46]([NH:48][C@H:49]([C@H:54]([CH2:56][C:57]([NH:59][C@H:60]([C:65]([NH:67][C@H:68]([C:75]([NH:77][C@H:78]([C:84]([O:86]C)=[O:85])[CH2:79][CH2:80][CH2:81][CH2:82][NH2:83])=[O:76])[CH2:69][C:70]1[N:74]=[CH:73][NH:72][CH:71]=1)=[O:66])[C@H:61]([CH2:63][CH3:64])[CH3:62])=[O:58])[OH:55])[CH2:50][CH:51]([CH3:53])[CH3:52])=[O:47])[CH2:40][C:41]1[N:45]=[CH:44][NH:43][CH:42]=1)=[O:37])[CH2:29][C:30]1[CH:35]=[CH:34][CH:33]=[CH:32][CH:31]=1)=[O:26])=[O:22])[CH2:14][CH2:15][CH2:16][NH:17][C:18](=[NH:20])[NH2:19])=[O:11])[CH2:3][CH2:4][CH2:5][NH:6][C:7](=[NH:9])[NH2:8].N>O>[NH2:1][C@H:2]([C:10]([NH:12][C@H:13]([C:21]([N:23]1[CH2:90][CH2:89][CH2:88][C@H:24]1[C:25]([NH:27][C@H:28]([C:36]([NH:38][C@H:39]([C:46]([NH:48][C@H:49]([C@H:54]([CH2:56][C:57]([NH:59][C@H:60]([C:65]([NH:67][C@H:68]([C:75]([NH:77][C@H:78]([C:84]([OH:86])=[O:85])[CH2:79][CH2:80][CH2:81][CH2:82][NH2:83])=[O:76])[CH2:69][C:70]1[N:74]=[CH:73][NH:72][CH:71]=1)=[O:66])[C@H:61]([CH2:63][CH3:64])[CH3:62])=[O:58])[OH:55])[CH2:50][CH:51]([CH3:53])[CH3:52])=[O:47])[CH2:40][C:41]1[N:45]=[CH:44][NH:43][CH:42]=1)=[O:37])[CH2:29][C:30]1[CH:35]=[CH:34][CH:33]=[CH:32][CH:31]=1)=[O:26])=[O:22])[CH2:14][CH2:15][CH2:16][NH:17][C:18](=[NH:19])[NH2:20])=[O:11])[CH2:3][CH2:4][CH2:5][NH:6][C:7](=[NH:8])[NH2:9]

Participants

4

Reactant 2

HCID 222Ammonia2×HCID 178455900HCID 178455900, compound

Product 1

HCID 178455899HCID 178455899, compound

Reagents, catalysts & solvents 1

HCID 962WaterSolvent

Procedure

80 mg of H-Arg-Arg-Pro-Phe-His-Sta-Ile-His-Lys-OMe (Example 7) are dissolved in 1.5 ml of H2O and the pH value is adjusted to 5.5 by the addition of 0.1N NH3. 8 μl of a 1% aqueous trypsin solution are added and the pH value is maintained at 5.5 by the addition of 0.1N NH3 by means of a pH stat. After approximately 20 minutes the absorption of base is complete, whereupon 350 μl of glacial acetic acid are added and the whole is heated for 2 minutes in a boiling water bath. The solution is concentrated to dryness, the residue is dissolved in 2 ml of 0.05N AcOH and, in order to separate off the inactivated enzyme, pumped through a column (φ=1.2 cm, length=25 cm) of Biogel P-10 polyacrylamide gel, exclusion limit at a molecular weight of 20 000; manufacturer: Bio-Rad, Richmond, Calif., USA). The fraction having a molecular weight matching the peptide mentioned below, which is ascertained by UV detection, is concentrated to a small volume and lyophilised, yielding H-Arg-Arg-Pro-Phe-His-Sta-Ile-His-Lys-OH; Rf (D)=0.12; Rf (M)=0.07; Rf (N)=0.05.

Workup

  1. addition8 μl of a 1% aqueous trypsin solution are added
  2. customAfter approximately 20 minutes the absorption of base
  3. additionare added
  4. temperaturethe whole is heated for 2 minutes in a boiling water bath
  5. concentrationThe solution is concentrated to dryness
  6. dissolutionthe residue is dissolved in 2 ml of 0.05N AcOH and, in order
  7. customto separate off the inactivated enzyme
  8. concentrationis concentrated to a small volume

Sources & evidence

ORD record
ord-5f149b6cb5c74165a3c631c61ded346a
Source dataset
uspto-grants-1986_06
Patent number
US04595677

Reaction notes

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