Sulfuric acid magnesium salt (1:1) → 3-Carbamoyloxymethyl-7-amino-3-cephem-4-carboxylic acid

HRID 616524Notes

Reaction equation

HRID 616524 reaction equation
[NH2:1][CH:2]1[C:14](=[O:15])[N:4]2[C:5]([C:11]([OH:13])=[O:12])=[C:6]([CH2:9][OH:10])[CH2:7][S:8][C@H:3]12.P([O-])([O-])([O:18][C:19](=O)[NH2:20])=O.S([O-])([O-])(=O)=O.[Mg+2]>[Cl-].[Mn+2].[Cl-].C(O)(=O)C>[NH2:1][CH:2]1[C:14](=[O:15])[N:4]2[C:5]([C:11]([OH:13])=[O:12])=[C:6]([CH2:9][O:10][C:19](=[O:18])[NH2:20])[CH2:7][S:8][C@H:3]12

Participants

6

Reactant 3

HCID 24083Sulfuric acid magnesium salt (1:1)HCID 3423467Carbamic acid, monoanhydride with phosphoric acid, ion(2-)HCID 129054807-Amino-3-hydroxymethyl-3-cephem-4-carboxylic Acid

Product 1

HCID 127847593-Carbamoyloxymethyl-7-amino-3-cephem-4-carboxylic acid

Reagents, catalysts & solvents 2

HCID 10313134Manganese(2+) dichlorideCatalystHCID 176Acetic AcidSolvent

Procedure

7-amino-3-hydroxymethylceph-3-em-4-carboxylic acid (40μg), [14C] carbamoyl phosphate (150μg, 7.5μCi), magnesium sulphate (20μg), manganese chloride (20μg) and pipes buffer (0.5μmole, pH 6.75) were incubated with O-transcarbamoylase as described in Example 9(i). After incubation the mixture was acidified with glacial acetic acid (10μl), clarified by centrifugation and the supernatant dried by rotary evaporation. This material was taken up in acetate/formate buffer [glacial acetic acid (5ml), formic acid (0.2ml) in methanol/water (40:60 v/v, 1 liter), pH adjusted to 3.9 with 4N sodium hydroxide] (30μl) and a portion (10μl) was applied to a Partisil 10 SAX column (250mm×4.6mm) and eluted with the same buffer at 1000 psi (1ml/minute). The U.V. absorption at 260nm and radioactivity of the eluate were monitored. A new U.V. absorption peak was detected after 12 minutes which also contained radioactivity. This peak corresponded to that obtained with authentic title compound. The conversion of cephalosporin starting material into title compound was 13%.

Workup

  1. customthe supernatant dried by rotary evaporation
  2. washeluted with the same buffer at 1000 psi (1ml/minute)
  3. customabsorption at 260nm and radioactivity of the
  4. customabsorption peak

Sources & evidence

ORD record
ord-3504b47618494521b483dcea2b24dec4
Source dataset
uspto-grants-1978_02
Patent number
US04075061

Reaction notes

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