DE2435619A1 - N-ACYL DERIVATIVES OF DESALANYLTETAIN AND THE PROCESS FOR THEIR PRODUCTION - Google Patents

Description

Politechnika Bdanska, '24 ·. July 1974

Gdansk / Poland "WZ / '

PL 216? ;

j N-acyl derivatives of '

Desalanyl tetaine and process for its, manufacture

The invention relates to N-acyl derivatives of desalanyl tetaine and a process for their preparation.

The antibiotic tetaine is produced by the strain of ύ Bacillus pumilus and has a number of interesting biological properties. It is characterized by a very high antibacterial activity, the range of activity of which includes numerous gram-positive as well as gram-vegetative and many other microorganisms.

A particularly valuable property of tetain is its low toxicity for animal organisms. This is based on a specific mechanism of action that is used to inhibit the cell ?! - <· wand-murein synthesis of the microorganisms leads. This is brought about by inhibiting the incorporation of the first amino acid, ie alanine, into the N-acetylmuramic acid residue. Antibiotics that inhibit this enzymatic process are not yet known.

509809 / 1U0

The Investigation of a New Inhibition Mechanism in Murein Synthesis gives through the synthesis of structural analogues of Tetains New Opportunities for the Manufacture of Non-Toxic Chemotherapy Drugs. The making of such connections is also useful because the tetaine itself is sensitive to a whole series of enzymes, which are light cause its ineffectiveness in living organisms.

On the one hand, these enzymes lead to the hydrolysis of the peptide bond of the tetaine molecule between the carboxyl group of L-alanine and the amino group of the remaining part of the tetaine molecule, i.e. desalanyl tetain. Free desalanyl tetaine formed in this way is biologically inactive.

These enzymes, on the other hand, catalyze a number of unfavorable reactions that occur in the desalanyl tetaine pragment of the antibiotic take place, and which also lead to the formation of biological ineffective substances contribute.

N-acyl derivatives of desalanyl tetain as well as processes their production are not yet known.

The purpose of the present invention is the preparation of N-acyl derivatives desalanyl tetaine, as well as the creation of a process for their manufacture.

The inventive derivatives of dasalanyl tetain are characterized in that they contain an N-acyl radical consisting of an et -amino acid or a peptide, with the configuration D or L or DL, or a group which is divided into an amino

509809 / 1U0

group or peptide group can be converted. \

The process for the preparation of N-acyl derivatives of desalanyl tetain is based on the acylation of the amino group of desalanyl tetain or its derivatives, for example peptides such as tetaine, by N-protected amino acids or peptides or by other [ compounds who are capable; to form a peptide group, \ weeks following the protection of the functional group of the final product, if necessary, \ removed. I.

The desalanyl tetaine / DAT / is a mixture of two isomeric substances called desalanyl tetaine A and B which are made from tetaine. That is tetain itself
a mixture of two isomeric compounds designated as tetaine A and tetaine B.

Both mixture components have the same malfunctional group J pen and similar reactivities. Semi-synthetic derivatives; can be made in an identical manner from both components A and B; as well as from their mixture. ,

The inventive N-acyl derivatives have the advantage that
they have a higher resistance to the above
Have enzymes.

The inventive N-acyl derivatives and the method to
their preparation are illustrated by the following examples.

Example I.

15 mg desalanyl tetaine in a lyophilized powder form

509809 / 1U0 " ^Λ

we'rden suspended in 1 ml of dry dimethylformamide. After stirring, 15 mg of D-Z-Z-ot-aminophenyl-acetic acid chloride-i hydrogen chloride added. After the reagents have been stirred for 1 hour, 5 ml of ether are added and the precipitate which has separated out is added is filtered off. The main product obtained is D - <? C aminophenylacetyl desalanyl tetaine. l

Example II. I

15 ml desalanyl tetaine A suspended in 0.1 ml dimethylformamide 1 are with Io mg D-Z-Z- * -aminophenyl acetic acid-N-carboxyan- ' hydride implemented. The pH is raised with N-methylmorpholine brought about 9. After stirring for 1 hour, the solvent becomes

evaporated in vacuo. The main product obtained is D - <* -Amino- ' phenylacetyl desalanyl tetaine.

_t j

Example III.

A solution of 1.5 mg of L-alanine-N-carboxyanhydride in 0.5 ml of tetrahydrofuran ^{1 is} added to 3.7 mg of desalanyl tetain B in 0.5 ml of water. After 2o hours of reaction at O ⁰ C, the Mi-! Schung evaporated in a vacuum. The main product obtained is crude tetaine B.

Example IV.

A solution is added to 3.7 mg of desalanyl tetaine in 0.5 ml of water of 2.4 mg of L-phenylalanine-N-carboxyanhydride in 0.5 ml Dimethylformamide. After a reaction time of 20 hours at 0 ° C., the mixture is evaporated in vacuo. As the main product L-phenylalanyl-desalanyl tetaine is obtained.

Example V.

To 37 mg desalanyl tetain in 5 ml Michaelis buffer / pH 8.2 /

509809/1 UO

give ^{a solution of 21.2 mg at 0} ° C. with vigorous stirring! L-alanine-N-carboxyanhydride in 5 ml tetrahydrofuran. After a reaction time of 20 hours at ⁰ ° C., the mixture is evaporated in vacuo and the crude tetaine obtained is purified on a chromatographic silica gel column with chloroform / methanol / water / 16: ^: 0.5 /. Yield: lo%.

Example VI.

21.2 mg of L-alanine-N-carboxyanhydride are added to 37 mg of desalanyl tetaine B in 5 ml of buffer / "pH Io /" at ⁰ ° C. with vigorous stirring. After 5 minutes, the pH is adjusted to 7 and the tetaine obtained B isolated according to Example V. Yield: 78 ^

Example VII.

A solution of 3 mg of L-alanine-N-carboxyanhydride in 1 ml of tetrahydrofuran is added to 10 mg of Tetain A-LyopillSat in 1 ml of water. After 20 hours of reaction at ⁰ ° C., the mixture is evaporated in vacuo The main product obtained is L-Alanyltetain B.

Example VIII. J

A solution of Ώ-αί -aminophenylacetic acid-N-carboxy anhydride in 1 ml of dimethylformamide is added to 10 mg of Tetain A lyophillate in 3 ml of dimethylformamide. After a reaction time of 20 hours at ⁰ ° C., the mixture is evaporated in vacuo. The main product obtained is D- oi- aminophenylacetyl tetaine A.

Example IX.

A solution of 1.8 mg of L-phenylalanine-N-carboxyanhydric is added to 10 mg of tetaine lyophilisate in 1 ml of Michaelis buffer / pH 8.2 /. in 1 ml of tetrahydrofuran. After a reaction time of 20 hours at ⁰ ° C., the mixture is evaporated in vacuo. The main product obtained is L-phenylalanine.

509809 / 1U0 ₆

Example X.

500 mg Benzoylglycylphenylphenylalanylalaninv ^ gestures in 10 ml of tetrahydrofuran was suspended and 0.11 ml of N-methylmorpholine added, cooled to -10 ⁰ C and treated with 0.13 ml ¹ J- isobutyl chloroformate. After vigorous stirring at this temperature for 15 minutes, a solution of 200 mg desalanyl tetaine in 5 ml Michaelis buffer / 8.2 pH / is added dropwise. The reaction is carried out at ⁰ ° C. for 3 hours. After this time has elapsed, tetrahydrofuran is removed in vacuo, and after the pH has been adjusted to 6, the reaction product, ethyl acetate, is extracted. After evaporation of the solvent, the residue is suspended in a phosphate buffer of pH 7.6 with the addition of 25 mg of chymotropsin. The reaction mixture is stirred at 30 ° while the reaction is monitored by chromatography. After the substrate has completely disappeared, the solution is brought to pH 5 and the precipitate is centrifuged. The sorption of the crude tetaine is carried out with tyrosine-deactivated carbon, followed by desorption with a 6% aqueous butanol solution. The extract is concentrated in vacuo and purified on a chromatographic silica gel column with butanol / water / methanol / 10: 10: 1 / as the top layer. Reaction yield: approx. HO%. \

^{Other N-acyl-:} derivatives of desalanyl tetain can be prepared by the methods mentioned above, which in the after- '

are summarized in the following table. !

... 7 509809/1 UO

N Desalanyl derivatives of the manufacturing

Tetains method

1. D-phenylalanyl-desalanyl tetaine - IV

2. D-alanyl-desalanyl tetaine · V

3. Glycyl desalanyl tetaine III

4. L-tyrosy-desalanyl tetaine III

5. L-valyl-desalanyl tetaine III

6. L-norvalyl-desalanyl tetaine. III

7. L-leucyl-desalanyl tetaine IV

8. L-isoleucyl-desalanyl tetaine IV

9. L-norleucyl-desalanytetaine IV

10. β-Alanyl-desalanyl tetaine III

11. e ^ -Aminoisibutyryl-Desalanyltetaine III

12. L-ot-aminobutyryl-desalanyl-tain IV

13. L-cX-aminophenylacetyl-desalanyl tetaine IV

14. O-Benzylseryl-desalanyl tetaine IV

15. S-Benzylcysteinyl-Desalanltetaine IV

16. Sarcosyl-desalanyl tetaine "III

17. L-N-methyl-alanyl-desalanyl tetaine III

18. N-phenylglycyl-desalanyl tetaine IV

19. D-Phenylalanyl-Tetaine VII

20. D-Alanyl-Tetaine VIII

21. Glycyl-Tetaine "VII

22. L-tyrosyl-tetaine ■ IX

23. L-valyl-tetaine IX

24. L-norvalyl-tetaine IX

25. L-leucyl-tetaine IX

26. L-isoleucyl-tetaine IX

27. L-norleucyl-tetaine IX

28. β-Alanyl-tetaine x IX

509809 / 1U0 ••• ⁸

Desalanyl derivatives of tetaine

Manufacturing methods

29. et -Aminoisobutyryl-tetaine

30. L-c * -aminobutyryl-tetaine

31 · L-c ^ -aminophenylacetyl tetaine

32. O-Benzylseryl-tetaine

33. S-Benzylcysteinyl-tetaine 3 ^. Sarcosyl tetaine

35. L-N-methylalynyl-tetaine

36. N-phenylglycyl-tetaine

IX

IX

VIII

VIII

VIII

IX

IX

VIII

509809/1140

All substances produced were thin-layer paper and biochromatographically tested.

In addition, the enzyme cleavage products were and paper chromatography in the presence of standards.

509809/1 UO

Claims (2)

24356 HJ 2L--, July 1974-WZ / Pl 2167 claims 1. N-acyl derivatives of desalanyl tetain, characterized that the acyl group is a ci'-amino acid residue or peptide residue is the one absolute configuration D, L or DL, or one Has grouping that is converted into an amine or peptide group can be. 2. Expires for the production of derivatives according to claim 1, because » characterized in that the amino group of desalanyl tetaine or its derivatives, for example, peptides such as tetaine with N-protected amino acids or peptides, or with others Compounds which can form a peptide bond is acylated, after which the protective group is optionally known Way is removed from the final product. 509809 / 1UO