CS216583B1 - A method of separating the D, L-S-benzylpenicilamin optical isomers - Google Patents

Abstract

For the separation of enantiomers of S-benzylpenicillamine, insoluble forms of the enzyme penicillin-amidohydrolase can be advantageously used, e.g. the enzyme anchored on B. megaterinium cells or by means of mutually bound E. coli cells having preserved penicillin-amidohydrolase activity. The essence of this method of separation is that L-N-phenylacetyl-S-benzylpenicillamine is allowed to react with an insoluble form of penicillin-amidohydrolase in an aqueous suspension at pH 6 to 8, and the resulting mixture of L-S-benzylpenicillamine and D-N-phenylacetyl-S-benzylpenicillamine is separated in a known manner.

Description

The present invention provides a process for resolving the isomers of D, L-S-benzylpenicillamine, i.e., β, 4-dimethyl-S-benzylcysteine by insoluble forms of penicillin-emidohydrolaay.

It has been shown that for D, L-N-phenylacetyl-S-benzylpenicillamine, it is possible to selectively cleave the phenylacetyl group only from the L-form by incubation and penicillin aminohydrolase (Ch. A.o. 209 633).

Furthermore, it has been found that insoluble forms of this enzyme can be advantageously used for the resolution of the enantiomers of S-benzyl-penicilsmin, e.g. amidohydrolase activity (a.oe 203607). Both of these modifications allowed rapid recovery and reuse of the enzyme.

The underlying principle of this process is that ae D, LN-phenylacetyl-S-benzylpenicillamine is reacted with an insoluble form of penicillin aminohydrolaay in aqueous suspension at pH 6-8 and the resulting mixture of LS-benzylpenicillamine and DN-phenylacetyl-S-benzylpenicillamine is separated. in a known way.

The separation of racemic S-benzylpenicillamine is further illustrated in the Examples.

Example 1

To a solution of N-phenylacetyl-S-benzyl-D, L-penicillamine (21.5 g; preparation see The Chemiatry of PeniciBin. Princeton Univ.Presa, Princeton 1949, p.472) in a mixture of water (176 mL)., 1 mol /

1-NaOH (63 ml) and 0.2 mol / L Na-phosphate buffer pH 7.5 (50 ml) were added modified E. coli cells and foamyillin-amidohydrolase activity (15 g; enzyme activity:

g of wet mass released from penicillin G 77 (µmol) 16-aminopenicillanic acid in 1 min).

The resulting pH of the suspension was 7.3. The mixture was stirred with a magnetic stirrer at 37 ° C and the progress of the reaction was monitored by high performance liquid chromatography. After a 120 point incubation, the cells were collected by filtration through a cloth, washed with water and used for the next session. Further, the solution was treated in the same manner as described in FIG. a.o. 209633. 3.1 g (43

S-benzyl-L-penicillamine of m.p. 190-192 ° C, + 96.4 ° (c 0.2 1 mol / l-dCl). For C 7 gHH NONOgS (239.3) calculated: 60.22% C, 7.16% H, 5.85% N; Found: C, 60.35; H, 7.22; N, 5.78.

The procedure was repeated with the same cell fraction. 3.4 g (47%) of the product with m.p. 191-193 ° C, + 97.6 ° (? 0.2, 1M-HCl); Found: C, 60.27; H, 7.17; N, 5.85.

Example 2

To a solution of N-phenylacetyl-S-benzyl-D, L-penicillamine (0.5 g) in a mixture of water (2.5 mL) and 1 mol / L

1-NaOH (1.5 ml), whose pH was adjusted to 7 by the addition of 0, 21 1-NaH ₂ PO ₄ (0.5 ml), was added a suspension of the enzyme anchored to B. megaterinium cells (containing 3.7 ml of its own). mass). The reaction was carried out at 37 ° C for 24 hours, the anchored enzyme was filtered off and the solution was worked up as in Example

1. 122 mg (73%) of S-benzyl-L-penicillamine were obtained at bt187-190 ° C and β47ρ + 93.2 ° (ο 0.2 1 mol / l HCl; found: 69.82% C) H 7.19% N 5.89%

Claims (1)

Method for separating optical isomers of D, LS-benzylpenicillamine, characterized in that D, LN-phenylacetyl-S-benzylpenicillamine is reacted with an insoluble form of penicillin-emidohydrolane in aqueous suspension at pH 6-8 and a blend of LS-benzylpenicillamine and DN-phenylacetyl. -S-benzylpenicillamine is separated by known methods.