FI68810C - NYA SAOSOM MELLANPRODUKTER ANVAENDBARA AMINOSYRADERIVAT - Google Patents

Description

L 68810

New amino acid derivatives useful as intermediates

The invention relates to novel amino acid derivatives, their salts and optically active isomers. The compounds of the invention are useful as intermediates in the preparation of compounds with valuable biological and pharmaceutical properties. Such compounds are those amino acid derivatives which are prepared according to Finnish patent application 751256.

The amino acid derivatives of the invention have the general formula I.

R -'-HN-CH-CO-A1 (CH.) I 2 n ^ CO-JJ - ((: H) - (CH2) t-BJ- (I) R R520 wherein A is hydroxyl, optionally p C 1-6 aralkoxy substituted with alkoxy or nitro or - (NH-CHR 6 -CO) -Y, 6 ly where R is hydrogen or C 1-6 alkyl, Y is hydroxyl or 1-4-C 1-4 alkyl ~ alkoxy and r is an integer from 1 to 10, B is -SO 2 OH, or -OPO (OH) 2, R is hydrogen or C 1-4 alkyl, R "^ is hydrogen, C 1-4 alkoxycarbonyl or optionally substituted with one or more of halogen, alkoxy and nitro substituted C 1-6 alkoxycarbonyl, R is hydrogen or carboxyl, n is 1 or 2, and t is 1 or 2, wherein A 1 is not hydroxyl if R 1 is hydrogen.

Acta Chemica Scandinavica 21 (1967) 35 1593-1604, page 1602 discloses a compound of formula I, 2,68810 wherein B1 is -SSR "; Ν, Ν'-bis - (β-p-toluenesulfonyl) -L-glutamylcystamine.

All the compounds of the invention are novel and the following are preferred compounds: 5 / ((. * - benzyl) -glutamyl-β-taurine, [N-carbobenzyloxy- (*-benzyl) -aspartyl-β-taurine, / ( ok (4-benzyl) -aspartyl-? Homotaurine, N-carbobenzyloxy- (N-benzyl) -aspartyl-N-cholineamine phosphate, N-carbobenzyloxy- (? -Benzyl) -glutamyl-N-homo-taurine, -carbobenzyloxy- (N-benzyl) -glutamyl-N-cholineamine phosphate, N-carbobenzyloxy- (N-benzyl) -glutamyl-N-cysteic acid, N-carbobenzyloxy- N-cysteamide) -glutamyl) -oC.7-Glycine ethyl ester, N-carbobenzyloxy- (N-taurine) glutamyl-N-glycine ethyl ester, N, N-carbobenzyloxy - (N-taurine) glutamyl - N - glycine-n-ethyl ester, ((N '- (taurine)) - L-glutamyl-O-N-glycine and (N-carbobenzyloxy- (OL-benzyl) -L-glutamyl) -N-taurine.

One structural feature common to all compounds of the formula I is that they have a dicarboxylic acid substituted in the β-position or also a derivative substituted in its other positions, which is bonded at its β-carboxyl group by an acid amide bond to a primary or secondary alkylamine in which the dicarboxylic acid in addition to the various substituents on the alkyl side chain, its derivative has a group which is strongly acidic in nature and which is in the ^ position.

The compounds of formula I can be prepared as follows: 3,68810

(a) for the preparation of compounds of general formula I having a sulphonyl, sulphonyloxy or phosphoryloxy group, compounds of general formula V

5 R ^ HN-CH-CO-A1 (CH) I 2 n 2 CO-N- (CH) - (CH_) -BZ (V) I 5 2 t

R R

Wherein A, R, R, R, n and t are as defined above and B is -SH, -SOOH, -OH, p-toluenesulfonyloxy, halogen or -SO 2 R 10, wherein R 2 is C N-alkoxy or aralkoxy, oxidized or hydrolyzed, reacted with an alkali metal or alkali metal hydrogen sulfite, esterified with sulfuric acid or phosphoric acid or a derivative thereof, or

b) for the preparation of compounds of general formula I in which B 1 is -SO 2 OH, compounds of general formula VI

R1-HN-CH-CO-A1 (CH) CO-N-R12 (VI) k1 25 II 12 wherein R, A and n are as defined above, R is hydrogen or N-alkyl, and R is an alkali metal, is subjected to reacting with alkali metal salts of haloalkylsulfonic acid, or

C) for the preparation of compounds of general formula I in which B 2 is -SO 2 OH, the compounds of general formula VII are oxidized

_ .. TT "

6881 O

4 r7-hn-ch-co-a4 «PVn CO-N- (CH) - (CH.) -S- (VII) 1 I 5 ^ t

R R

5 __ __ 2 4 wherein A is hydroxyl, aralkoxy, preferably benzyloxy, substituted aralkoxy, preferably p-methoxybenzyloxy or p-nitrobenzyloxy, 7 R is C 1-4 alkoxycarbonyl or optionally halogen, alkoxy with one or more substituents and nitro-substituted C 1-4 aralkoxycarbonyl and 5 "

R, R, n and t have the same meaning as above, or d) compounds of general formula XVI

R 7 N-C-COOH

I I

0 = C- (CH2) n (XVI) 7 20 wherein R and n are as defined above are reacted with compounds of the formula H2N - (<p ») -‘ cH2> t-E1

Rb 25 1 5.

wherein B, R and t are as defined above, or with salts thereof, or compounds of general formula XVIII

30 R7-HN-CH-CO-A4 (CH.) (XVIII) 2 n CO-A7 5 6 8 310 4 7 7 wherein A, R and n are as defined above and A is hydroxyl, p-nitrophenyloxy, pentachlorophenyloxy or C 2-4 alkoxycarbonyl, is reacted with compounds of general formula XX 5 HN- (CH) - (CH).-B1 II 5 2 t RR (XX)

wherein B 1, R 2, R 5 and t are as defined above, or 10 f) compounds of general formula XXI

r7-hn-j; h-co-a4 (CH ~) | 2 n ^ CO-N- (CH) - (CH2) -B (XXI) | lc 2 t

15 R R

4 1 5 7 wherein A, B, R, R, R, n and t are as defined above, a protecting group is cleaved from the amino group, or g) compounds of the general formula 4 1 5 7 20 XXI above, wherein A, B, R, R, R, n and t have the same meaning as above, the protecting group is removed from the α-carboxyl group, or

h) for the preparation of compounds of general formula XXIII

25 R1-HN-CH-CO-A8 (CH) I 2 n, CO-N- (CH) - (CH) -B (XXIII)

I I ς 2 t R RJ

30

wherein A8 is - (NH-CHR6-CO) -Y and B1, R, R1, R5, n and t are as defined above, the β-aminocarboxylic acid-containing peptide derivatives activated at the β-position are reacted with compounds having the general formula. va XXIV

6881 O

6 HN - (CH) - (CH2) -B4 R R5 (XXIV) 45 wherein B is -SO4 -OH, -OPO (OH) 2 or -SH and R and R are as defined above, and optionally the mercapto group is oxidized, and the resulting compounds are converted to their salts or liberated from their salts and / or the compounds are prepared as optically active isomers by using optically active reaction components in an optional reaction step or by performing racemate resolution.

In process variant (a), the compounds of general formula V are preferably reacted with perforic acid or a mixture of glacial acetic acid and hydrogen peroxide or the compounds are heated with an aqueous alkali metal sulphite solution.

In process variant (b), the compounds of general formula VI are preferably reacted with bromoethanesulfonic acid or bromopropanesulfonic acid starting metal salts.

In process variant (c), the compounds of general formula VII are preferably reacted with a mixture of glacial acetic acid and 30% hydrogen peroxide.

In process variant (d), the compounds of the general formula XVI, preferably pyroglutamic acid, are preferably reacted with taurine or homotaurine.

In process variant (e), compounds of general formula XVIII, preferably 30 N-carbobenzyl 1-aminocarboxylic acid 1 * -bentyl 1 - [- p - nitrophenyl ester in a mixture of pyridine and water, are preferably reacted with compounds of general formula XX. preferably with taurine, N-methyltaurine, homotaurine, choline phosphate or cystamic acid.

Il 7 6381 0

In process variant (f), the carbobenzyloxy group is preferably removed from the compounds of general formula XXI, preferably from N-carbobenzyloxy- * benzyl ester derivatives, preferably with hydrogen bromide in glacial acetic acid.

In process variant (g), the ester group of the compounds of general formula XXI, preferably N-carbobenzyloxy-o-4-benzyl ester derivatives, is preferably removed by saponification.

In the preparation of salts of the compounds of the general formula I, the compounds of the general formula I are preferably reacted with alkali metal or alkaline earth metal hydroxides or carbonates or with organic bases.

The invention is described in more detail in the following examples.

Example 1; N-L-glutamyltaurine (a) 40.85 g (0.11 mol) of carbobenzyloxy-L-glutamic acid oxy-benzyl ester (Liebigs Annalen 20 655 (1962) 200) are dissolved in 500 ml of acetonitrile.

The solution is cooled to -15 ° C, while preventing the effect of air humidity. 15.4 ml (0.11 mol) of triethylamine are then added dropwise to the solution with stirring, then 15.4 ml (0.11 mol) of chloroformic acid isobutyl 25 ester. The reaction mixture is stirred at -15 ° C for 40 minutes, then 28 ml (0.2 mol) of triethylamine are added, followed by 11.26 g (0.05 mol) of cystamine hydrochloride and finally 250 ml of acetonitrile. The mixture is then stirred for a further 2 hours at -15 ° C and 4 hours at room temperature.

At the end of the reaction time, the mixture is evaporated at 30 ° C in vacuo. The residue is dissolved with stirring and cooling in 200 ml of ice water, and the mixture is again evaporated in vacuo at 35 ° C. The residue is taken up in water (250 ml) and ethyl acetate (500 ml) into a separatory funnel and the organic phase is separated. The organic phase is shaken successively with water (250 ml), 5% sodium carbonate solution (2 x 250 ml), 1N hydrochloric acid (2 x 250 ml) and finally with water (250 ml) (shaking the sodium phase with sodium carbonate solution 5 can be made by acidified with hydrochloric acid and shaken with ether to recover about 5 g of unreacted carbobenzyloxy-L-Glu-aminic acid-6 * benzyl ester). The ethyl acetate phase is dried over anhydrous sodium sulfate and then evaporated to dryness in vacuo at 30 ° C. A thick oily residue is obtained which soon crystallizes into a crystalline mass. The crude product (40-42 g) is recrystallized from a mixture of ethyl acetate (100 ml) and ether (170 ml). 29.3 g of N, N1-bis / N-carbobenzyloxy - '- (<* - benzyl) -L-glutan-15-yl] -cystamine are obtained, m.p. 91-92 ° C.

Elemental analysis for C 44 H 50 N 4 O 2 Sq = ^ 59.05) Calculated: C 61.52%, H 5.89%, N 6.52%, S 7.46% Found: C 60.85%, H 5.91%, N 6.61%, S 7.72%.

(b) 25.77 g (0.03 mol) of the Ν, Ν'-bis-20 / N-carbobenzyloxy-E- (c, benzyl) -L-glutamyl-7-cystamine obtained above are dissolved in 75 ml of glacial acetic acid. . A freshly prepared mixture of 30% hydrogen peroxide (75 ml) and glacial acetic acid (225 ml) is added dropwise over 15 minutes to an ice-cooled solution. After the addition, cooling is stopped and the reaction mixture is stirred for 4 hours at room temperature. It is then evaporated to dryness in vacuo at 30 ° C. The oily product is first dried in a desiccator with phosphorus pentoxide, then with solid potassium hydroxide. 28.5 g of carbobenzyloxy-N- (N-benzyl) -L-Glu-30 tamyl taurine are obtained. The crude product can be used without purification to prepare β-L-glutamyl taurine.

(c) 26.32 g (55 mmol) of the carbobenzyloxy-N- (N-benzyl) -L-glutamyltaurine prepared above are dissolved in 50 ml of glacial acetic acid. Add 4 l to the solution

6881 O

9 moles of hydrogen bromide dissolved in 50 ml of glacial acetic acid. Rapid evolution of carbon dioxide can be observed. The reaction mixture is allowed to stand for 2 hours at room temperature, then evaporated to dryness in vacuo at 30 ° C. The oily residue 5 is dissolved in 170 ml of water and shaken with ether (5 x 70 ml). The aqueous phase is evaporated to dryness in vacuo at 35 ° C. 20.42 g of (N-benzyl) -L-glutamyl litaurine are obtained, which is recrystallized from 90% ethanol, m.p. 204-208 ° C.

Rf (n-butanol-pyridine-glacial acetic acid-water 15: 10: 3: 12) = 0.53 Rf (n-butanol-glacial acetic acid-water 4: 1: 1) = 0.39.

(d) 20.42 g of the β- (oC-benzyl-L-glutamyl taurine obtained above) are dissolved in 150 ml of 1N potassium hydroxide solution and allowed to stand at room temperature for 4 hours to 15 hours, then taken up in the H + step. A 2 x 100 cm statue filled with Dowex 50 x 2 (Fluka, 100-200 mesh) is eluted with water, 300 ml of eluate are collected from the beginning of the elution and evaporated to dryness in vacuo at 35 ° C. 8-10 ml of water and about 100 ml of ethanol, filtered, washed with alcohol and dried to give 13.7 g of -L-glutamyl taurine The crystalline product is recrystallized from 80% aqueous ethanol to give 9.79 g of pure product corresponding to a yield of 70% of N, N 1 -bis- / N-carbobenzyloxy- (R-benzyl) -L-glutamyl-7-cystamine.

Example 2: N-L-glutamyl taurine (a) 529 mg (1.1 moles) of the carbobenzyloxy-benzyl) -L-glutamyl taurine prepared in Example 1b are dissolved in 5 ml of 1 N potassium hydroxide and the solution is allowed to stand at room temperature for 4 hours. The solution is then shaken with ether (3 x 3 mL). The aqueous phase is transferred to a 1 x 20 cm column filled with Dowex 50 x 2 and eluted with water. Collect 50 ml of the solution, which is evaporated to dryness in vacuo at 35 ° C. Crude carbobenzyloxy-β-L-glutamyl taurine is obtained, which is purified by paper electrophoresis at pH 6.5. The relative mobility compared to cysteic acid is 1.05.

(n-butanol-pyridine-glacial acetic acid-water 15: 10: 3: 12) = 0.57. (b) The carbobenzyloxy-β-L-glutamyl litaurine obtained above is dissolved in 2 ml of glacial acetic acid containing 4 moles of hydrogen bromide. The mixture is allowed to stand at room temperature for 10 and a half hours, then evaporated to dryness in vacuo at 35 ° C. The residue is triturated several times with ether Q i and separated from it by decantation. The resulting ε-L-glutamyl taurine is recrystallized as described in Example 3.

Example 3: N-L-glutamyl taurine 5.79 g (13.1 mmol) of the carbobenzyloxy- (N-benzyl) -L-glutamyl taurine prepared in Example Ib are dissolved in a mixture of ethanol (100 ml) and water (25 ml) and hydrogenated with shaking in the presence of 10% palladium on carbon-20 catalyst (0.5 g). The catalyst is suitably added in two portions of 0.25 g. When no more hydrogen bonding occurs, the solution is filtered and then evaporated to dryness in vacuo at 30 ° C. The oily residue is dried in a desiccator over phosphorus pene-25 oxide. 3.1 g of N-L-glutamyl taurine are obtained, which is very soluble in water and insoluble in alcohol. Addition of a little water and alcohol in small portions causes the product to crystallize. The crystalline product melts at 202-204 ° C.

In the IR spectrum, the character groups have the following maxima:

Onh * x OH 3600 - 2400; NH (amide) 3315; V CO (carboxyl) 1758; ^ CO (amide) 1666; 0nh * 1576; 0 SO- 1296, 1031 cm -1.

6881 O

11

The crude product is crystallized several times from 80% ethanol. 2.02 g of pure final product are obtained in a yield of 66% with respect to N, N'-bis- / N-carbobenzyloxy- (OL · -benzyl) -L-glutamyl-7-cystamine. The melting point of the pure product 5 is 219-220 ° C. + 14 ° (water, c = 1.02).

The relative mobility relative to cysteic acid in paper electrophoresis at pH 6.5 is 0.73 and at pH 1.8 is 0.53.

R t (n-butanol-pyridine-glacial acetic acid-water 15: 10: 3: 12) = 0.19 10 Analysis for C 1 H 2 O 2 S (M = 254.27)

Calculated: C 33.07%, H 5.55%, N 11.02%, O 37.75%, S 12.61% Found: C 33.15%, H 5.76%, N 10.94% , O 37.53%, S 12.17%.

Example 4: N-L-glutamyltaurine 5.42 g (11 mmol) of carbobenzyloxy-L-glutamic acid - (N-benzyl) -p-nitrophenyl ester (Chem. Ber.

96, 204, 1963) is dissolved in 50 ml of pyridine. The solution was cooled to 0 ° C and then added dropwise within a period of half an hour to 20 with vigorous stirring, a solution of 1.25 g (10 mmol) of taurine in 20 ml of water. 3.08 ml (22 mmol) of triethylamine are then added to the mixture, and cooling and stirring are stopped. The mixture is allowed to stand at room temperature for 72 hours, then evaporated to dryness in vacuo. The residue is dissolved in 50 ml of water and 1N hydrochloric acid is added to the yellow solution until the color disappears, the solution is shaken with ether (10 x 50 ml) to remove p-nitrophenol. The aqueous phase is evaporated to dryness in vacuo. 6.9 g of carbobenzyl-30-oxy-M-benzyl) -L-glutamyltaurine triethylammonium salt are obtained.

This material is catalytically hydrogenated as described in Example 3a and the solvent is removed by evaporation in vacuo. To remove triethylamine, the material is dissolved in a small amount of water and the solution is taken up in a Dowex 50 x 2 12 6881 0 column (2 x 40 cm). Elute with water. The eluate is taken up in about 120 ml and evaporated to dryness in vacuo at 35 ° C. The product is crystallized and isolated as described in Example 3. 1.72 g of 5β-L-glutamyltaurine are obtained, corresponding to a yield of 60% with respect to taurine.

In the IR spectrum, the character groups have the following maxima:

OnH * x OH 3600 - 2400; VNH (amide) 3315; V / CO (carb-10-carboxyl) 1758, -00.00 (amide) 1666; in 1576; 0 SO ~ 1296, 1031 cm_1.

Example 5: fr-L-glutamyltaurine (a) 0.48 g (1 mmol) of carbobenzyloxy-L-glutamyl- (β-nitrophenyl ester) -glycine ethyl ester 15 (Acta Chim. Acad. Sci. Hung. 65, 375, 1970) is dissolved in 6 ml of ethyl acetate. The solution is cooled with ice water. At 0 [deg.] C., 0.08 g (1 mmol) of cysteamine in 1 ml of dimethylformamide are added dropwise to the solution, then 0.14 ml (1 mmol) of triethylamine. A precipitate slowly forms.

The reaction mixture is kept in ice water for some time and then at room temperature for 24 hours, finally diluted with a 1: 1 mixture of ether and ethyl acetate. The precipitate is separated by centrifugation and washed first with a 4: 1 mixture of ether and ethyl acetate, then several times with ether. After drying over sulfuric acid, the precipitate is washed 3 times with 1N hydrochloric acid, 2 times with water, 2 times with saturated sodium hydrogen carbonate solution and finally 2 more times with water and then dried in vacuo over sulfuric acid. 0.35 g of carbobenzyl-30-oxy-β-L-glutamyl- (β-cysteamine) -glycine ethyl ester is obtained, corresponding to a yield of 85%.

Analysis for C 18 H 19 N 2 O 2 N 2 S (M = 411.4) i 6868 0

Calculated: C 62.6%, H 6.1%, S 7.8% Found: C 53.4%, H 6.5%, S 7.7%. In the infrared spectrum, the character groups have the following maxima: 5 Y NH 3310; Y C = O (COOEt) 1748; Y C = O (Z) 1690; Y C = O (amide) 1655 cm -1.

100 mg of carbobenzyloxy-06-L-glutamyl- (N, N-cysteamine) -glycine ethyl ester are dissolved in 2 ml of glacial acetic acid, and 0.5 ml of 30% hydrogen peroxide is added to the solution.

The reaction mixture is allowed to stand for 4 hours under ice-water cooling. The progress of the reaction is monitored by paper electrophoresis. After dilution with water, the reaction mixture is lyophilized. The final product is obtained as a foam. 0.11 g of carbobenzyloxy-O-L-glutamyl- (β-taurine) -glycine-15-ethyl ester (95%) are obtained.

(b) 100 mg of the carbobenzyloxy-L-glutamyl- (4-taurine) -glycine ethyl ester prepared above are dissolved in a mixture of trifluoroacetic acid (1 ml) and concentrated hydrochloric acid (1 ml). The solution is kept at 35 ° C for 3 hours in 20 sealed bomb tubes. The solution is then evaporated to dryness in vacuo, the residue is digested several times with ether and then n-hexane is then evaporated to dryness again. A white amorphous substance is obtained which proves to be uniform in the electrophoresis study and which gives 25 positive ninhydrin spots. The product is N-L-glutamyl- (N-taurine) -glycine, yield 0.06 g (88%).

Analysis for C 9 H 8 N 2 O 2 S (M = 311.3)

Calculated: S 10.3% Found: S 10.0%.

30 In the infrared spectrum, the character groups have the following maxima: 3100 (broad); )> OH (COOH) 3200 (broad); C = O (COOH) 1730; )> C = O (amide-I) 1680; ^ C = O (amide-II) 1560; p S = 0 (SO 2 OH) 1220 (intense); S = 0 (SO 2 O) 35) ^ 1045 cm -1 (intense).

i4 6 8 81 0

Hydrolysis: 20 mg is kept in 1 ml of 6 N hydrochloric acid in a fused glass tube at 105 ° C for 24 hours. After cooling, a sample of the solution is electrophoresed at pH 1.8. The solution contains glutamic acid, glycine and taurine.

100 mg of the? -L-glutamyl- (p-taurine) -glycine prepared above are dissolved in 25 ml of 0.2 molar Arammonium hydrogen carbonate buffer, pH 8.5. To the solution is added 1 mg of carboxypeptidase A dissolved in 0.5 to 10 ml of water (enzyme manufacturer: Serva, Heidelberg).

The mixture is kept for 24 hours in a thermostat at 37 ° C, then lyophilized. From the dry lyophilisate, β-L-glutamyl taurine and glycine can be detected. Pure glutamyl taurine is obtained by electrophoresis or chromatography on a Dowex 50 column.

Example 6: N, N-Glutamyl taurine (a) 0.47 g (1 mmol) of carbobenzyloxy-o-CL-glutamyl (N, N-p-nitrophenyl ester) glycine methyl ester are dissolved in 6 ml of pyridine. - 0.125 g (1 mmol) of taurine in 2 ml of water, then 0.28 ml (2 mmol) of triethylamine are added in such small portions that the solution remains clear at all times, and the reaction mixture is allowed to stand at room temperature for 3 days. in vacuo, an oil is obtained which is thoroughly digested in ether, then in petroleum ether and finally dried in vacuo over sulfuric acid to give carbobenzyloxy-? (, - L-glutamyl- (r-taurine) -glycine methyl ester.

(b) 100 mg of the carbobenzyloxy-30 ° C-L-glutamyl- (1H-taurine) -glycine methyl ester prepared above is reacted at room temperature with a 2N hydrochloric acid acetic acid solution (4 ml) until the whole is dissolved (about 30 minutes). The resulting clear solution is poured into 30 ml of ether and left in a cool place for 35 days. The precipitate formed is centrifuged, washed several times with ether and then dried in vacuo over potassium hydroxide, sulfuric acid and phosphorus pentoxide. As the electrophoretic study shows, the hydrobromide 5 of ° C-L-glutamyl - (N-taurine) -glycine methyl ester 5 is practically pure.

(c) The salt obtained above is reacted for 3 hours with 1N sodium hydroxide solution (2 ml) while cooling with ice water. The progress of the hydrolysis is monitored by electrophoresis. The reaction mixture is subjected to 10 ion exchanges with 10 ml of Dowex 50 H +, then lyophilized. Since the electrophoresis still shows impurities, the product is crystallized several times from aqueous ethanol until a sufficient degree of purity is reached. 40 mg (59%) of α-L-glutamyl- (N-taurine) -15 glycine are obtained.

In the infrared spectrum, the character groups have the following maxima: NH 3310; VnH ^ 3100 (broad); C = O (COOH) 1730; ^ 0 = O (amide-I) 1650; >> C = O (amide-II) 1570; > * S = 0 1220 20 (intense), 1045 cm 2 (intense).

(d) 100 mg of ° C-1-glutamyl- (Λ · taurine) -glycine prepared above are dissolved in 25 ml of 0.2 molar ammonium hydrogen carbonate buffer at pH 8.5. To the solution is added 1 mg of carboxypeptidase A dissolved in 0.5 to 25 ml of water (enzyme manufacturer: Serva, Heidelberg).

The mixture is kept in a thermostat for 24 hours at 37 ° C, then lyophilized. From the dry lyophilisate, N, L-glutamyl taurine and glycine can be detected. Pure N-L-glutamyl taurine is obtained by electrophoresis or chromatography on a Dowex 50 column.

Example 7: N, N-Glutamyl homotaurine (a) 1.083 g (2.2 mmol) of carbobenzyloxy-L-glutamic acid (N-benzyl) -β-nitrophenyl ester are dissolved in 6 ml of a 2: 1 aqueous pyridine mixture. To solution 35 is added first 278 mg (2 mmol) of homotaurine, then 16 6881 0 0.59 ml (4.2 mmol) of triethylamine. The yellow solution is allowed to stand at room temperature for 72 hours and then evaporated to dryness in vacuo. the oily residue is dissolved in water, neutralized with hydrochloric acid and then extracted to remove 5 p-nitrophenol in a continuous extraction apparatus with ether for 8 hours. The aqueous phase is evaporated in vacuo. 1.68 g of carbobenzyloxy-benzyl) -L-glutamylhomotaurine are obtained.

(b) The entire amount prepared above (1.68 g) is dissolved in 10 ml of 50% aqueous ethanol, then 0.3 g of 10% palladium on carbon is added to the solution, and hydrogen is passed through the suspension for 4 hours. The solution is then filtered and evaporated in vacuo. The residue is dissolved in 1-2 ml of water and the solution is taken up in a Dowex 50 x 2 column (1 x 35 cm) to remove triethylamine in H + form. Let's live with water. The eluate is taken up in 50 ml and then evaporated to dryness. The residue contains 440 ml of N-L-glutamyl homotaurine, corresponding to a yield of 82%. Electrophoresis at pH 6.5 shows 20 small amounts of partly neutral, partly acidic substances (homotaurine or glutamic acid). The product can be purified, for example, by preparative electrophoresis.

In both pH 6.5 and pH 1.8 electrophoresis, the substance migrates to the cathode. Its relative mobility to cysteic acid is 0.68 at pH 6.5 and 0.50 at pH 1.8.

(n-butanol-pyridine-glacial acetic acid-water 15: 10: 3: 12) = 0.19 In the IR spectrum, the characteristic groups have the following maxima: 30 P-NH 3345; > * NH *, OH 3200 - 2200; V CO 1740; TP CO-NH 1645; y SO "1240, 1190, 1150, 1038; (f -NH 1570, 1535; SO ~ 590 cm -1).

Example 8: r-L-glutamyl-N-methyltaurine (a) 1.083 g (2.2 mmol) of carbobenzyloxy-L-35 glutamic acid (benzyl) p-nitrophenyl ester 17 6881 0 is reacted with N-methyltaurine (278 mg, 2 mmol). ) as described in Example 7a. 1.59 g of carbobenzyloxy-N '- (N-benzyl) -L-glutamyl-N-methyltaurine are obtained.

5 (b) The entire amount of the material prepared above (1.59 g) is hydrogenated from the catalyst as described in Example 7b. 423 mg of L-glutamyl-N-methyltaurine are obtained, corresponding to a yield of 79%.

The substance migrates at both pH 6.5 and pH 1.8 in paper-10 electrophoresis per anode. The relative mobility compared to cysteic acid is 0.68 at pH 6.5 and 0.49 at pH 1.8.

(n-butanol-pyridine-glacial acetic acid-water 15: 10: 3: 12) = 0.16. Example 9: N-Glutamyl-L-cysteic acid (a) 2.87 g (6.6 mmol) of carbobenzyloxy-L-glutamic acid (<> C-benzyl) -β-p-nitrophenyl ester are dissolved in 20 ml. 1.25 g (6 mmol) of L-cysteic acid monohydrate dissolved in a mixture of water (17 ml) and pyridine (17 ml) are added to the solution, and a further 2.6 ml (18.6 mmol) of triethylamine are added, which The reaction mixture is then allowed to stand at room temperature for 72 hours, then the solution is evaporated in vacuo at 30 [deg.] C. The residue is dissolved in 20 ml of water, acidified with concentrated hydrochloric acid and then shaken with ether (15 x 10 ml). evaporated in vacuo at 35 [deg.] C. to give carbobenzyloxy-[(C-benzyl) -L-glutamyl-L-cysteic acid.

(b) The product prepared above is dissolved in 20 ml of water, 0.3 g of 10% palladium-on-carbon is added, and hydrogen is passed through the suspension for 3 hours. The reaction mixture is worked up as described in Example 7b. N-L-glutamyl-L-cysteic acid is obtained, melting at 187 ° C. By paper chromatography, the relative mobility to cysteic acid is 1.21 35 at pH 6.5 and 0.54 at pH 1.8.

18 6881 0 In the IR spectrum, the characteristic groups have the following maxima:}) NH (amide) 3423; OH 3300 - 2200; j) CO (carboxyl) 1748, 1712; J> CO (amide) 1627; j) NH (amide) 1527; 5 .Pso ”1221, 1109, 1043; ($ SO ~ 525 cm "1.

Analysis for cqhi4N2 ° 8S = 298.28)

Calculated: C 32.86%, H 4.73%, N 9.39%, S 10.75% Found: C 33.06%, H 5.28%, N 9.05%, S 11.11% .

Example 10: L-glutamylcholamine phosphate 10 (a) 1.083 g (2.2 mmol) of carbobenzyloxy-L-glutoiminoic acid (oC-benzyl) p-nitrophenyl ester are dissolved in 6 ml of a 2: 1 mixture of pyridine and water. To the solution is added first 282 mg (2 mmol) of choline phosphate (U.S. Patent No. 2,730,542) and then 0.87 ml (6.2 mmol) of triethylamine. The reaction mixture is allowed to stand at room temperature for 72 hours, then evaporated in vacuo. Further work-up is carried out as described for carbobenzyloxy-r- (CZ * -benzyl) -L-glutamyl homotaurine (Example 7a). 1.25 g of carbobenzyloxy-N- (O-benzyl) -glutamyl-colamine phosphate are obtained.

(b) The product obtained above (1.25 g) is catalytically hydrogenated to remove protecting groups. Hydrogenation and ion exchange purification are performed as described for the preparation of N, L-glutamyl homotaurine (Example 7b). 470 mg of N-L-glutamylcholamine phosphate are obtained, corresponding to a yield of 91%. However, as can be seen in paper electrophoresis, the product contains about 15 to 20% choline phosphate as an impurity. As a cleaning method, e.g. electrophoresis.

In paper electrophoresis, the substance migrates at both pH 6.5 and pH 1.8 per anode. The relative mobility compared to cysteic acid is 0.75 at pH 6.5 and 0.36 at pH 1.8.

35 Rf (n-butanol-pyridine-glacial acetic acid-water 15: 10: 3: 12) = 0.18.

19 6881 0

Example 11: β-L-aspartyltaurine (a) 526 mg (1.1 mmol) of carbobenzyloxy-L-aspartic acid (CX-benzyl) - β-p-nitrophenyl ester (Chem. Ber. 97, 1789, 1964) are dissolved To 5 ml of pyridine.

The solution is cooled to 0 ° C and then a solution of 125 mg (1 mmol) of taurine in 2 ml of water is added in small portions, followed by 0.28 ml (2 mmol) of triethylamine. The reaction mixture is allowed to stand at room temperature for 48 hours, then evaporated in vacuo.

Dissolve the residue in 5 ml of water and initially add 1N hydrochloric acid to the yellow solution until the color disappears, shake the solution with ether (10 x 5 ml) to remove p-nitrophenol. The aqueous phase is evaporated in vacuo. 478 mg of carbobenzyloxy - ((β-benzyl) -L-aspartyl-15-litaurine are obtained.

(b) The entire amount of product obtained above is dissolved in 6 ml of 50% aqueous ethanol. 100 mg of 10% palladium-on-carbon are added to the solution, and the mixture is then hydrated for 4 hours by passing hydrogen gas through it.

After filtration and evaporation in vacuo, the triethylamine is removed from the product by the ion exchange method described for the preparation of N, L-glutamyl homotaurine (Example 7b). 17 2 mg / μl of L-aspartyl taurine are obtained, corresponding to a yield of 71%. The product contains some taurine as an impurity, which can be removed e.g. by electrophoresis.

In paper electrophoresis, the substance migrates at both pH 6.5 and pH 1.8 per anode. The relative mobility to cysteic acid is 0.77 at pH 6.5 and 0.58 at pH 1.8.

(n-butanol-pyridine-glacial acetic acid-water 15: 10: 3: 12) = 0.16.

Example 12: N-L-aspartylhomotaurine (a) 526 mg (1.1 mmol) of carbobenzyloxy-L-aspartic acid (δ-benzyl) - (δ-nitrophenyl ester) and 35,139 g (1 mmol) of homotaurine are reacted in Example 6881. 20 as described in Cat 11. Carbobenzyloxy- [b- (δC-benzyl) -L-aspartyltaurine is obtained.

(b) The product obtained above is catalytically hydrogenated as described in Example 7b. 203 mg (84%) of 5- (b-L-aspartyl homotaurine) are obtained.

In paper electrophoresis, the substance migrates at both pH 6.5 and pH 1.8 per anode. Compared to cysteic acid, the relative mobility is 0.72 at pH 6.5 and 0.53 at pH 1.8.

Rf (n-butanol-pyridine-glacial acetic acid-water 15: 10: 3: 12) = 0.17.

Example 13: (R) -L-aspartylcholamine phosphate (a) Carbobenzyloxy-L-aspartic acid (N, N-benzyl) -N-p-nitrophenyl ester and cholamine phosphate are reacted with each other as described in Example 10. Carbobenzyloxy-N- (N, N-benzyl) -L-aspartylcholine phosphate is obtained.

(b) The material obtained above is catalytically hydrogenated as described in Example 7b. <3-L-aspartylcholine phosphate is obtained.

In paper electrophoresis, the substance casts at both pH 6.5 and pH 1.8 per anode. Compared to cysteic acid, the relative mobility is 0.81 at pH 6.5 and 0.40 at pH 1.8.

(n-butanol-pyridine-glacial acetic acid-water 15: 10: 3: 12) = 0.14.

Claims (13)

Intermediates for the preparation of therapeutically useful amino acid derivatives of the general formula 1'h2n-ch-co-oh (CH) j 2 n (i ·) CO-N-CH- (CHJ -B1; | £ t. 10. r5 5 wherein R is hydrogen or C 1-4 alkyl, R is hydrogen or carboxyl, R 1 is -SO 2 OH or -OPO (OH) 2, n is 1 or 2 and t is 1 or 2, and pharmacologically acceptable salts thereof and novel 15 amino acid derivatives useful in the preparation of optically active isomers, characterized in that they have the general formula I R1HN-CH-CO-A1 (CH-) I 2 n / j) 20 I 1 '^ 1 CO-N- (CH) - (CHJ. -Q ti 2 t where A 1 is hydroxyl, optionally substituted in the p-position by alkoxy or nitro substituted C-, O-aralkoxy or - (NH-CHR CO) ,, -Y, 6 / -yr where R is hydrogen or C 1-6 alkyl, Y is hydroxyl or 1-4 η C 1-4 alkoxy and r is an integer from 1-10, B is -SO 2 OH or -OPO (OH) 2, R is hydrogen or C 1-4 alkyl, R 1 is hydrogen, C 1-4 alkoxycarbonyl or optionally substituted by one or more halogen, alkoxy and nitro substituents C 1-6 aralkoxycarbonyl, 5'-y R is hydrogen or carboxyl, n is 1 or 2 and 35 is 1 or 2, wherein A 1 is not hydroxyl if R 2 is hydrogen, their salts and optically active isomers. _______ - ίγ ^ ----- 22 6381 0 Amino acid derivative according to Claim 1, characterized in that it is β-benzylglutamyl-Y 7 -taurine, a salt thereof or an optically active isomer thereof. The amino acid derivative woman according to claim 1, characterized in that it is N-carbobenzyloxy- (α-benzyl) -aspartyl-β-taurine, a salt thereof or an optically active isomer thereof. Amino acid derivative according to Claim 1, characterized in that it is / (β-benzyl) -aspartyl-β-halothiourine, a salt thereof or an optically active isomer thereof. Amino acid derivative according to Claim 1, characterized in that it is N-carbobenzyloxy- (<β-benzyl) -aspartyl-β-choline phosphate, a salt thereof or an optically active isomer thereof. 5 6 8 81 0 Amino acid derivative according to Claim 1, characterized in that it is N-carbobenzyloxy- (cK-benzyl) -glutamyl-N-homotaurine, a salt thereof or an optically active isomer thereof. Amino acid derivative according to Claim 1, characterized in that it is (N-carbobenzyloxy-M-benzyl) -glutamyl-cholamine phosphate, a salt thereof or an optically active isomer thereof. Amino acid derivative according to Claim 1, characterized in that it is (N-carbobenzyloxy-u-benzyl) -glutamyl-cysteic acid, a salt thereof or an optically active isomer thereof. Amino acid derivative according to Claim 1, characterized in that it is N-carbobenzyloxy- (Ϋ-ky steamide) -glutamyl-α-glycine ethyl ester. Amino acid derivative according to Claim 1, characterized in that it is N-carbobenzyloxy- (**-taurine) -glutamyl-<7-glycine ethyl ester. Amino acid derivative according to Claim 1, characterized in that it is N-carbobenzyloxy - (N-taurine) -glutamyl-* 7-glycine methyl ester. 23 6 881 0 Amino acid derivative according to Claim 1, characterized in that it is N, N-taurine-L-glutamyl-β-glycine or a salt thereof. Amino acid derivative according to Claim 1, characterized in that it is N-carbobenzyloxy- (N-benzyl) -L-glutamyl-N-taurine, a salt thereof or an optically active isomer thereof. 6881 0 24