FI65990B - FOERFARANDE FOER FRAMSTAELLNING AV NYA TERAPEUTISKT ANVAENDBARA AMINOSYRADERIVAT - Google Patents

Abstract

1504541 Compositions containing amino acid derivatives CHINOIN GYOGYSZER ES VEGYESZETI TERMEKEK GYARA RT 28 April 1975 [29 April 1974 26 March 1975] 17608/75 Heading A5B [Also in Divisions C2 and C3] Cosmetic and pharmaceutical compositions contain (I) and/or salts thereof as active ingredient wherein A<SP>1</SP> stands for hydroxy, C 1-4 alkoxy, cycloalkoxy, aralkoxy, substituted aralkoxy, aryloxy, substituted aryloxy or a group of the general formulae: wherein R<SP>14</SP> is hydrogen, C 1-4 alkyl or aralkyl, R<SP>6</SP> is hydrogen, C 1-5 alkyl, aralkyl, hydroxysubstituted aralkyl, heteroaralkyl or a group of the general formula: Y is hydroxy, amino, alkylamino, dialkylamino, C 1-4 alkoxy or aralkoxy, and r is an integer of from 1 to 10 or an average polymerization grade of up to 2000, B<SP>1</SP> is a group of the formulae -SO 2 OH, -OSO 2 OH, -O-PO(OH) 2 or -S-S-R<SP>11</SP>, wherein R<SP>11</SP> is C 1-4 alkyl, aralkyl or aryl or a residue obtained when removing group B<SP>1</SP> from the general formula (I), R stands for hydrogen, C 1-4 alkyl or aralkyl, R<SP>x</SP> stands for hydrogen or halogen, R<SP>1</SP> stands for hydrogen, C 1-4 alkyl, aryl, aryl having a nitro or alkoxy substituent, aralkyl, substituted aralkyl, alkoxycarbonyl, cycloalkoxycarbonyl, aralkoxycarbonyl, substituted aralkoxycarbonyl having, e.g. a halogen, alkoxy, nitro, phenylazo or alkoxyphenylazo substituent, unsubstituted or substituted aryloxycarbonyl, acyl, arylsulfonyl or group (wherein R<SP>6</SP> has the same meanings as defined above and p is an integer of from 1 to 10 or an average polymerization degree of up to 2000), R<SP>2</SP> stands for hydrogen, C 1-4 alkyl, or aralkyl or R<SP>2</SP> and R<SP>2</SP> may each stand for a -CO- group and form a ring through an o-phenylene, alkylene or -CH=CH- group, R<SP>3</SP> stands for hydrogen, carboxy or carbalkoxy, R<SP>4</SP> stands for hydrogen, halogen, C 1-4 alkyl or hydroxy, R<SP>5</SP> stands for hydrogen, halogen, C 1-4 alkyl, carboxy, carboxamido, carbalkoxy or carboaralkoxy, m is 1, 2 or 3, n is 1, 2, 3 or 4, s is 0, 1, 2, 3 or 4, and t is 1, 2 or 3.

Description

I, .- ^ 71 ra, .... KU RELEASE PUBLICATION

[B] O1) UTLÄGGNI NGSSKR1FT 6 5 9 90 C Patent granted 10 03 1934 ^ 'Patent se-cyclelat ^' r ~ * '(51) KHk? / 00, C 07 F 9/09 FINLAND —FINLAND (2.1) P »t« rttlh * l <«mut - Pitenunuttknlng 751256 (22) HakumltpUvi - Anteknlngudtg 25.0 ^ .75 (23) Alku cloud - Glltlghatadag 25.0 ^ .75 (41) Tullut JulklMksI - Bllvlt offamlig 30.10.75

Patent · 14 registered administrations ....,. . .....

*. . (44) Date of Nihtlvikalpano's hearing. -. "

Patent- och reglttentyrelaen Anattkan utlagd och uti.skrtft «n pubiicörad 30. Oh. oh (32) (33) (31) Requested «until then — Begird priorkst 29. OA. Jk 26.03.75 Hungary-Hungary (HU) FE-928, CI-1558 (71) CHINOIN Pharmaceutical Chemicals Termgkek Gyära RT., T6 u. 1-5, Budapest IV, Hungary-Hungary (HU) (72) Läszlö Feuer, Budapest, Ärpäd Furka, Budapest, Ferenc Sebestyen, Budapest, Jolän Hercsel, Budapest, Erzsebet Bendefy, Budapest,

Hungary-Hungary (HU) {7k) Oy Kolster Ab (51+) Method for the preparation of new therapeutically useful amino acid derivatives - Förfarande för framställning av nya tera-peutiskt användbara aminosyraderivat / s' s

The invention relates to a process for the preparation of new therapeutically useful amino acid derivatives of general formula (I)

H ^ N-CH-CO-OH

(<TH2 n I 2 n 100-γ- ^ Η- (CH2) tB (I) R R5 wherein R is hydrogen or C1-4alkyl, is hydrogen or carboxyl, is “SC2OH, -OSC2OH or -OPCMOH 1 / n is an integer of 1 or 2 and t is an integer of 1 or 2 to prepare pharmacologically acceptable salts and optically active isomers thereof.

2 c 5 99 0

A common structural property of all compounds of general formula (I) is that they contain an o <-substituted dicarboxylic acid or a derivative thereof also substituted at other positions which is linked via an O) -carboxyl group by an acid amide bond to a primary or secondary alkylamine further substituted by various substituents. in the alkyl side chain, a group having a strong acidic character at the W position.

Of the compounds according to the invention, special mention should be made of β-L-glutamyltaurine of the formula (XXIV)

H ^ N-CH-COOH,

“I

CH2 (XXIV) CH2 co-nh-ch2-ch2-so2oh and having a direct or immediate broad spectrum of therapeutic and prophylactic effects on pathological changes due to damage to "AGAS" (aerobiospheric genetic adaptation system).

To illustrate the concept of "AGAS", the following is a list of the major tissues and organs that make up the rumen system.

(a) All biological interfaces in contact with the outdoor air as the biosphere (skin and skin tissue, cornea and conjunctiva, oral and pharyngeal cavities, respiratory tract and lungs); (b) bones and limbs (tubular and fungal bones, ball joints, synovial membrane, skeletal muscle); (c) organs involved in the regulation of the ionic economy (transepithelial transport system: intestinal lobe and renal tubules): (d) artificial dentition (attached to the roots of the tooth pit) for grinding food; (e) auditory, olfactory and auditory organs.

Thus, the compounds of the invention have a beneficial biological or therapeutic effect on the organs or tissues of the AGAS system listed herein.

3 63990

The compounds of the invention further affect the following functions associated with the AGAS system: radiation protection, wound healing, primary activating effect on primary tissue, protection against the ever-increasing risk of infection and aging of the skin and mucous membranes (lysozyme production of moist mucosa, protection against infections caused by viruses and fungi.

The compounds of the invention counteract the ever-increasing effects of life stress (e.g. climatic effects, large differences in day and night temperatures, increased risk of vulnerability) by stabilizing adaptation syndrome while combating peripheral tissue damage caused by goiter cocoids (e.g. connective tissue damage, bone marrow damage, bone marrow damage Development of immune homeostasis (increased ability of the body to recognize which cells are the body's own and which are not).

FI patent 51,429 discloses a method for isolating and purifying a hormone concentrate that activates elemental scaffold and raises blood silicon content.

GB 1 092 185 discloses a therapeutic composition comprising the calcium salt of choline phosphate and potassium and / or magnesium aspartate.

The process according to the invention for the preparation of new amino acid derivatives of the general formula I, their pharmacologically acceptable salts and their optically active isomers is characterized in that a) a compound of the general formula (II)

R 1 HN-qH-CO-OH

<CHo> n I 2 .1 CO-N-CH- (CH0) .- B (II) I Is 2 1

R R

wherein R is (1) C 1-4 alkoxycarbonyl or (2) C 7-9 aralkoxycarbonyl optionally substituted by one or more halogen atoms or one or more alkoxy or nitro groups, and R, R, B, n and t is as defined above, the protecting group R is cleaved by acidolysis or hydrolysis, or 65990 b) for a compound of general formula (III)

HoN-CH-CO-A2 2 i (CH2) n CO-N-CH- (CH2) t-B1 (III) I 15

R R

2 wherein A is (1) C 1-4 alkoxy, (2) g-aralkoxy optionally substituted in the p-position by alkoxy or nitro, or (3) a group of formula (b) (-NH-CH -CO) -Y (b) 1 · r 6 wherein R is hydrogen or C 1-4 alkyl, Y is hydroxyl or C. Alkoxy, 5 1 1 and r is an integer from 1 to 10, and R, R, B, n and t is as defined above, alkaline or enzymatic hydrolysis, acidolysis or hydrogenolysis is carried out, or c) a compound of general formula (IV) 7 3

R -HN- <j] H-CO-A

Wn is 1 CO-1 -CH- (CH 2) t B (IV) 3 wherein A is (1) C 1-4 alkoxy or (2) C 7-9 aralkoxy optionally substituted at the p-position with alkoxy or nitro, and R, R 7, R, B, n and t are as defined above, the protecting groups R and A are simultaneously cleaved by acidolysis, alkaline hydrolysis or hydrogenolysis, or d) a compound of general formula (V)

1 1 R -HN-CII-CO-A

j 2 'n CO-N-CH- (CH2) t'B2 (v) I! 3

R R

6 5 9 9 0 5 1

wherein R, R, n and t have the same meaning as above, A

is (1) hydroxy, (2) C 1-4 alkoxy, (3) C 1-4 aralkoxy optionally substituted in the p-position by alkoxy or nitro, or (4) a group of formula (b) wherein R 1, Y and r are as defined above, R is (1) hydrogen, (2) -N-alkoxycarbonyl, or (3) C 1-6 aralkoxycarbonyl optionally substituted with one or more halogen atoms, or 2 with one or more atoms - or with a nitro group, B is -SH, 10 -SOOOH, -OH, p-toluenesulfonyloxy, halogen or -SOnR, 10 *. wherein R is C 1-4 alkoxy or aralkoxy, is oxidized, or if the compound of formula (V) contains a sulfonic acid ester group, partial hydrolysis is performed, or if B is p-toluenesulfonyloxy or halogen, a compound of formula (V) is obtained react with alkali sulphite or alkali hydrogen sulphite to give compounds of general formula (I) in which B 1 is -SO 2 OH, or if the compound of formula (V) contains hydroxyl groups, it is esterified with sulfuric acid or phosphoric acid or a derivative thereof to give compounds of formula (I) containing a hydrogen sulphate or dihydrogen phosphate group and, if desired, cleavage of the protecting groups attached to or bonded to the β-amino and / or β-carboxyl groups, or e) for the preparation of compounds of general formula (I) in which B is a group containing a sulfonyl, a compound of the general formula (VT) R 1 -HN-CH-CO-A1 KVn CO-N-R 12 l · 3 11. 12 wherein R, A and n are as defined above, R is hydrogen 13 C 1-6 alkyl, and R is an alkali metal, is reacted with an alkali metal salt of a haloalkylsulfonic acid, followed by cleavage of any protecting groups attached to or attached to the N, amino and / or carboxyl groups, or f) reacting compounds of general formula (I). for the manufacture of. . 1 wherein B is -SO 2 OH, a compound of general formula (VII) r7-hn-ch-co-ä4 j i hVn! (VII,: CO-N-9H- (CH9) -Ξ ι I s 1 t; r R 'L_l 2 65990 4 wherein A is hydroxyl, aralkoxy or preferably benzyloxy, substituted aralkoxy, preferably p-methoxybenzyloxy- or p- nitrobenzyloxy, and R, R, R7, n and t are as defined above, are oxidized, followed by protecting groups, R1 and A4 are cleaved or ct) to prepare compounds of general formula (I) wherein B is -SO4OH, a compound of general formula (XXII)

O O

l! I: CN c jj N CH "xnm ^ 3 ...... c '|, (XXII) 0 <αΙ2> η where n is an integer 1 or 2 and M is sodium or potassium, is reacted with a compound having general formula (XXIII)

Hal - CH - (CH_). - SO,., OH

I 5 2 t 2 in R (XXIII) 5] R and T have the same meaning as in formula (I) and Hal is halogen, or a salt thereof, a compound of formula (XV) 9 o c il, ·> \ Λ \

I || M -PU

i /. N-CH-CH 2), -SO-OH (XV) i '5

O I A R

(CH _) '~

2 n O

S

wherein R ', n and t are as defined above, are partially hydrolysed with a dilute acid or base to give a compound of formula (XXIV) 0 | 1 N-CH-COOH (XXIV) Ί 7, ^. / ...... c (CH „) m | 2 n 0 CO-NH-CH- (CH.J, -SO-OH k 2 t 2

R

5 wherein R, n and t are as defined above, is reacted with hydrazine, or h) a compound of general formula (XVIII) B '-NH-CH-CO-A4 (CH 2) (XVIII> I 2. Π

CO-A,

65990 4 7 7 wherein Λ, R and n are as defined above and A is hydroxyl, p-nitrophenyloxy, pentachlorophenyloxy or t 1-4 -alkoxycarbonyloxy are reacted with a compound of general formula (XX) HN -9H- (CH2) t-B1 (XX) k r5 1 5 in which B, R, R and t have the same meaning as above, after which the protecting groups A and R7 are cleaved from the compound obtained, preferably the protecting group R is cleaved by acidolysis and the protecting group A is cleaved by alkaline hydrolysis in any order and, if desired, converted or liberated from the pharmacologically acceptable salt and / or isolated as its optically active isomers using an optically active starting material in an optional reaction step or by resolution of the racemate.

In the detailed description of the process alternatives according to the invention, mention should first be made of process variant h) suitable for the preparation of all compounds of the formula I, according to which the compounds of the general formula (I) are prepared by forming an acid amide bond. In this case, a derivative substituted in the amino or amino and A-carboxyl groups of the x-aminodicarboxylic acid by one or more protecting groups or also by other groups is attached via a β-carboxyl group to, for example, 2-aminoethanesulfonic acid, 3-aminopropanesulfonic acid or 2-phosphonoethanolamine. When forming an acid amide bond, various protecting groups come into play. The most suitable coupling method is the active ester method (monograph on the formation and selective deprotection and coupling methods: E. Schröder, K. Lubke: The Peptides, Vol. 1: Methods of Peptide Synthesis, Academic Press, 1965).

Compounds of general formula (I) are likewise obtained in process variant d), in which a d-aminodicarboxylic acid amide is first prepared in which the amine moiety has a functional group other than a strongly acidic functional group, for example a sulfhydryl or sulfinic acid function. From these compounds, the desired compounds are prepared by oxidation. If the functional group is a halogen atom or a ρ-toluenesulfonyl group (J. Chem. Soc. 824, 1964), substitution with alkali sulfite or alkali hydrogen sulfite gives compounds of general formula (I) which contain a sulfone group. Compounds of general formula (I) containing a hydrogen sulphate or dive-phosphate group are obtained from compounds containing hydroxyl groups by esterification. If the starting compound contains a sulfonic acid ester group, then the compounds of general formula (I) are obtained by partial hydrolysis under mild conditions.

Furthermore, it is possible to start from glutamine or asparagine or their substituted derivatives (with the exception of acid amides). In this case, according to process variant e), for example, one of the hydrogen atoms of the acid amide of the acid amide is substituted with sodium metal, and the compound thus obtained is converted by reaction with 2-bromoethanesulfonic acid salts and optionally cleaving the protecting groups of the general formula (1). compound. The starting materials used are u) -amides of -aminodicarboxylic acids, for example w-vinyl-1-amide or u./-amide, in which the amine component is a nitrogen-containing three-membered ring compound (aziridine).

In process variant g), an alkali salt of β-aminoglutarimide or β-aminosuccinimide is first formed and this is then alkylated on an imide nitrogen platform. As the alkylating agent, for example, 2-bromoethanesulfonic acid or β-bromopropanesulfonic acid can be used. The N-sulfoethyl or N-sulfopropyl derivative thus obtained is then hydrolyzed in a weakly alkaline medium to form most of the ring compound -amide. This can be purified by ion exchange chromatography to remove the prescribed amide.

The compounds of the invention act in part directly, in part through the metabolism of vitamin A, by producing metabolites with a stronger polar nature than vitamin A. This effect can be compared to the effect of parathyroid hormone on the renal tubular 25-hydroxycholecalciferol-14-hydroxylase enzyme. With this clarification, the broad pharmacological, biochemical and therapeutic effects of the compounds of the invention will be understood. The effect of the compounds is as follows: 9 65990 A) Vitamin A-like effect: a) Pharmacological and biochemical effects: labeled sulphates accumulate to a large extent in rat sword cartilage or chicken embryo lens, liver and lung tissue respectively: radiolabelled phosphorus accumulates to a large extent in rat sword; chondroitin sulfate synthesis promoting effect; a beneficial effect on wound healing or, respectively, experimental wound healing in rats and dogs caused experimentally by the use of cortisone; The potentiating effect of vitamin A in experimentally induced hypo- or hypervitaminosis in rats and chickens; smoothing the stress-causing effect of gastric ulcer in rats; the effect of platelet degranulation; lysozyme production enhancing effect; effect on trace element economy (silicon, zinc, copper, manganese, fluorine); promoting effect on epithelial formation; an elevating effect on alkaline phosphatase activity, potency in the formation of granuloma sacs caused by the local action of vitamin A, a very gentle course of the dose-potency curve, or a change in the sign of the dose at high doses, respectively; activating effect on the Golgi apparatus; promoting effect on goblet cell formation; an elevating effect on vitamin A content; b) Clinical-therapeutic effects: keratoconjunctivis sicca, Sjögren's syndrome; rhino-laryngo-pharyngitis sicca; chronic bronchitis; sinobronkiitti; mucoviscidosis; small children's structural lung diseases; paradentosis; susceptibility of skin and mucous membranes to viral and fungal infections; cortisone antagonistic effect; beneficial effect on the healing of surgical and mucosal wounds; erosion Colli; pruritus, diseases; decreased sense of smell and taste.

B) Effects that do not have the character of vitamin A.

(a) Pharmacological and biochemical effects; a temporary lowering effect on the blood sugar mirror; elevating effect on phosphate urination, lowering effect on serum phosphorus mirror; radio shielding effect; in labyrinth experiments with inactive animals, a lowering effect on the time required to reach the target; lowering effect on experimentally induced fluoride and cadmium poisoning; elevating effect on renal cyclic adenosine monophosphate depletion; smoothing effect on symptoms of experimentally induced pea poisoning; decreased histamine sensitivity; elevating effect on liver enzyme tyrosine aminotransferase activity.

b) Therapeutic effects; slight radiation damage; vitiligo; lihashypotonia; psychoenergic effect; beneficial effect on involutionary and gerontological conditions such as memory loss functions; arpeutumistaipumukset; ankylosing spondylosis; musculoskeletal disorders; sclerotic fundus; amyloidosis; morphea; fibrocystic mastopathy.

The beneficial therapeutic effects of the compounds of the invention are also apparent from the following therapeutic experiments.

Therapeutic experiments

As can be seen from the following experiments, the compounds of formula I have the following effects: they lower blood sugar, increase serum vitamin A levels, increase blood silicon levels and, together with vitamin A, promote implantation-induced granuloma formation in wound care.

The following compounds of formula I were tested in the experiments: β-L-glutamyl taurine (referred to as litoralone) * '-aspartyl-N-methyltaurine -aspartyl-homotaurine β-glutamylcholamine phosphate s

The results shown in Tables 1-6 are averages. The tables also show the standard error of the results and the number of determinations performed in parentheses. The significance of the differences between the results given by the reference and test substances is determined by the Student's t-distribution.

a) Effect of litoralon on serum vitamin A levels

table 1

Group Litoralone Serum vitamin A concentration ^ g / day ________P3 * __ I (comparison) - 28.3 j 0.7 (20) II 0.1 41.9 - 1.0 * (20) III 0.3 32.9 - 1.1 (20) IV _M__27.4 - 0.6 ___ (20) 65990 nx P <0.001 xx P <0.01

In the experiment, 20 rats, half male and half female (Sprague Dawlev) and weighing 180-200 g, were treated orally for eight days with the daily dose of litoral in aqueous solution shown in Table 1. On the ninth day, the animals were killed and bled. The vitamin A content of the obtained serum was determined by the method of Neeld and Pearson.

65990 (k) interaction of kitprion and vitamin A on granuloma formation induced by the implantation of cotton swabs CN 00 00 00 CO 00 --g —----- irL --- L -----. “...

cd o e c

0 -H

OfC o r— m t ^ · σ \ r- rHCUtP * · * ·

3 td E r- M (N (M CN

C> + 1 + 1 +1 +1 + | +1

Cd -H <N * + rr ΙΛ Π O

M3 m m U ^

B

+ j M: cd ta>

0) -H

• H Ud i— x— Ή Dj I I I - -

m \ O O

• rl (J tT

CM3 ΓΜ 0 0 ^

rH

O M

-X O Jt v-

-X -P -h tn I I - I I

3 ^ n! 3. o rH mj a 3 tn

td O

H C ------------------------------------------------- - c <

•B

X P

-h tn c ta -h tn • H · Η

E

td f-t tr> il cn (n i cn <j>

+> cd E jC

-h λ; υ> -h o 1 cd Ä <Oi <d. . ......________ ______________________________.

C

cd cd e n O-ι 4-1

3 3 ~ O

rH rH C W

• H Ή * H

cd Cd +>

+ J + J O X

MM3 ta ta -h

j> _> rH

: <D

E

C H

> 4 H H> H

PS H H H H>> * »____ ____ 13 65990

The differences were significant: between groups II and III P <0.05, between groups II and V P <0.001 and between groups V and VI P <0.01.

Granuloma formation was determined by the method of Lee and colleagues using male rats (Sprague - Dawley) weighing 110-120 g. The swabs were removed from the subcutaneous subcutaneous tissue after 10 days and dried at 65 ° C and weighed.

14 65990 c) The effect of l · itoralonln on the development of experimental latyrism

Table 3

Bone

Group

Ca P CSA CP

I: comparison 14,4ίθ, 24 6.7 ^ 0.21 0.9210.01 0.37 ^ 0.023 (12) II: ΒΑΡΝ 12.6 ± 0.26 5.5 ^ 0.32 0.7 ± 0 .06.26 ± 0.02 (12) 10 III: BAPN 10 mg 14.2 ^ 0.30 6.5-0.14 0.90 ^ 0.037 0.35 ± 0.022 (12) ♦ litoralone 0.3 ! Cartilage j Itver comparison 1.52-0.047 0.84 ^ -0.026 2.18 ± 0.088 1.06 ± 0.032 (12) fj II: BAPN 1Omg 1.33-0.062 0.62-0.051 1.47ίθ, 20 0.84ί0 , 047 (12) y III: BAPN 10 mg A. . . / l + litoralone 1.50 ^ 0.036 0.8110.0492.17-0.19 0.9910.048 (12) j 0.3 ^ g!

The differences between groups II and III are significant when P <0.01.

Male rats weighing 140-150 g (Sprague-Dawley) were used in the experiment. With 0.3 yugrn of synthetic Lito-raloia with a purity of 98%. Table 3 shows the amounts of Ca, P, CSA (chondroitin sulfoute) and CP (chondroprotein) determined from tibia and sword-like cartilage as a percentage of wet weight.

Ca was determined by the method presented by Pribil, P was determined by the method presented by Baginski and colleagues, and CSA and CP were determined by the method not already presented by Dittmann's colleagues.

15 05990 d) Effect of litoral on blood silicon

Table 4 ~ - i ”'” --Λ

Silicon concentration in the blood, mg / g

5 days 7 days of the test

Group at baseline after after I: comparison 0.103 ^ 0.003 0.096i0.00g 0.114-0.015 (6) II: litoralone, 5 μg 0.095 ^ 0.006 0.154 ± 0.012 0.151 ^ 0.004 (6) III: litoralone, 10 μg 0.103 ^ 0.007 0.170-0.006 x 0.171-0.006 (6) After 13 days 20 days 40 days after | ______ | I: comparison 0.117-0.011 0.126 ± 0.009 0.149 ^ 0.011 (6) i: II: litoralone, 5 jaq 0.182 + 0.006x 0.303-0.012XX 0.331-0.013XX (6)! 1 1

+ X + XX + XX I

III: litoralone, 10 μg 0.201-0.01 5 0.365-0.01 3 0.355-0.01 4 (6) j ί x significance: P <0.01 xx significance: P <0.001

The results obtained on day 13 are significant when P <0.01 and on day 20. Results obtained when P <0.001. The experiment was performed on male rabbits (breeding rabbits) weighing 2.5-3 kg. Rabbits were orally administered litoralone daily at the doses shown in Table 4. Silicon content was determined by the Gaubatz method from 5 ml blood samples taken from the ear vein.

ie 65990 e) Effect of litoralon on metamorphos in Dalmatian frogs.

Table 5

Group Body length, mm Tail length, mm

Comparison 45.9 ^ 0.2 31.4ί 0> 2 (6Q) Treated animals 40.2ί0.2χ 26.7i 0.2X (60) x Significance: P <0.001

30-day-old Dalmatian frog larvae with a body length of 20–25 mm were used in the experiment. The beginnings of the hind legs were already visible. For 30 days, the experimental group was placed daily in tap water with 0.5 μg litoralone per ml for 2 hours. The control group was similarly maintained in tap water to which no litoralone had been added. Frog fry were measured on day 30.

f) The effect of litoralon on blood sugar

Table 6 i Group I study II study i mg% mg%

Comparison 94 + 3.6 (10) 94-4.09 (10) Treated animals 82.4 ± 3.8 (10) 81-1.32 (10) i i____________

Significance: Study I: P <0.05, Study II: P <0.01.

65990

White male rats (CGY) weighing 160-180 g were used in the experiment. Animals were maintained on a standard diet. A blood test was taken on day 5 of the experiment after an 18-hour fast. Blood glucose assays were performed according to the Hultman method. Litoralone was administered orally daily at a dose of 1 μg / kg.

2. / 3-aspartyl-N-methyltaurine a) Effect on blood sugar

Control group 107mg%

Experimental animals S3mg%

Significance: P <0.05

There were 20 rats in both the experimental and control groups. Assays were performed after 18 hours of fasting. The test substance was administered orally as a solution for four days at a dose of 1 μg / kg animal.

b) Effect of increasing serum vitamin A levels

Dose ^ Ug / 200g animal Effect on vitamin A-oral concentration, __________i_______________ 0 8.5 5 11.0 1 11.5 0.3 12.5 j 0.1 16.1! 0.05 14.8 0.01 12.5 0.005 10.5

Significance: P <0.01

There were 20 male rats (Wistar) weighing 200-220 g in both the experimental and control groups. The trial period lasted 6 days.

65990 18 c) Effect on blood silicon

Group Blood concentration, mg / g

At the beginning of the 40 days 40 days after the test I ______ [comparison 0, 0,00ί 0,004 0,120 ± 0,010 0,154 + 0,015 j

| l (5 yug / day) 0.100 = ^ 0.005 0.31 5 ± 0.01 4XX 0.346 ± 0.015XX

i

XX

lii (10yug / day) 0.107 + 0.009 0.370 ± 0.119XX 0.360 ± 0.017 ί _______________ xx Significance: P <0.001

The result is significant from day 13 when P <0.01 and from day 20 when P <0.001. Male rabbits (breeding rabbits) weighing 2.5-3 kg were used in the experiment. The active substance was administered orally daily at the dose shown in the table. Silicon assays were performed by the method of Gaubatz (Gaubatz E., Klin. Wschrft. 14 (1935) 1753) on 5 ml of blood samples taken from the ear vein of an experimental animal.

19 65990 d) Effect of β-aspartyl-N-methyltaurine and vitamin A on the formation of granuloma in cotton-wool implantation

Group Dose | Dry granule- x! Vitamin A Active substance | animal weight, topically topically! OrallyJ m <? mg ^ μg ^ ug / day; _ ___- ____ ________1 .... 1— --——, - .....

I (comparison) i: I j II (comparison -! 55-3.4! j and Solvent) j j I | III 2 i I ~ I 66-12.5 i 1 i IV 2 0.1 i 67-2.6 1 i i! v - 0.1 79 ± 2.8:! i! I! ! i VI! 2; 0.1 1 96-4.4 1! ! ! ______j_________I________________i___________l_______! x Hoffman La Roche The differences are significant as follows:

Between groups II and III P <0.05 and between groups II and V P <0.01 and between groups V and VI P <0.01.

Granuloma formation was determined in male rats (Sprague-Dawley) weighing 110-120 g by the method of Leen and colleagues (Lee KH, Fu Ch.Ch., Spencer MK, Tong TG and Poon RJ, Pharm. Soc., 62 (1973)). ) 895). Subcutaneous implanted tampons were removed after 10 days, dried at 65 ° C and weighed.

/ 20 65990 3. / 3-asparyl-homotaurine a) Effect on blood sugar

Control group 105 mg%

Experimental animals 94 mg%

Significance, dose, experimental animals and experimental arrangements as in 2a above (β-aspartyl-N-methyltaurine).

b) Effect of increasing serum vitamin A levels

Dose yug / 200 g animal Effect on vitamin A-i oral concentration, i _____ / »g * _ | 0 8.6 and 5 12.0 1 13.5 j 1.3 14.0 0.1 15.8 0.05 15.0! | 0.01 12.0 | 0.005 10.0 l ________________ ί

Significance, number of experimental animals and experimental arrangements, etc. as in 2b above.

21 65990 c) Effect on blood silicon content Group j Blood silicon content, mg / kg i | The start of the exam - 20 days 40 days! moment after

Comparison! 0; 104 ± 0.009 0.134 ± 0.015 10.157 ^ 0.020 j '| [! + | + xx | , xx I (5 ug / day) I 0.094-0.007! 0.309-0.014 | 0.340 ^ 0.014 i i

i! . XX + XX

II (10 μg / day) '0.109-0.010 0.372-0.120 0.363-0.018

Significance, number of experimental animals, experimental arrangements and method of analysis as in 2 c above.

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Significance Number of test animals, test arrangements and method of analysis as specified in paragraph 2d above.

4. β-Glutamyl-cholamine sulphate a) Effect on blood sugar

Control group 104 mg%

Experimental animals 93 mg%

Significance, number of experimental animals, experimental arrangements and dosage as in 2a.

b) Serum vitamin A-increasing effect

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Significance, number of experimental animals, experimental arrangements and method of analysis as in 2b above.

65990 24 c) Effect on blood silicon content

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Significance Number of test animals, test design and method of analysis as in 1c above.

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26 6 5 9 9 0

In the administration of the compounds according to the invention, the treatment time varies greatly. Some diseases (e.g. rhino-laryngo-pharyngitis sicca) become asymptomatic at a daily oral dose of 3x5 μg within two weeks, other months (e.g. paradentosis, Sjögren’s syndrome) take 1-2 months to resolve symptoms, while others (e.g. Spondylosis ankylipoetiea) requires 1 / 4-1 / 2 years of treatment.

The compounds according to the invention can be prepared in a simple manner into various pharmaceutical, prophylactic, cosmetic or, respectively, veterinary preparations. The preparations may contain only one active ingredient or may contain a combination of active ingredients. The daily dose of pure active ingredient per kg of body weight is 50-500 ng and is divided into three single doses.

One tablet contains 2-20 [mu] g, preferably 10 [mu] g of active ingredient, in addition to biologically inert carriers (e.g. milk sugar, starch) and customary tableting aids (granulation and glidants, such as e.g. polyvinylpyrrolidone, gelatin, talc, magnesium stearate, aerosil, etc.).

Due to the very low dose, it is expedient to add the active ingredient to the tablet mass as a solution before granulation. In this way, the active ingredient is evenly distributed. The active substances are permanent and the tablets can therefore be marketed without notification of the expiry date. The active ingredient content of sustained-release tablets or spans capsules may be 10 to 20 μg. A suitable dose for injection preparations is 5 to 10 g per ampoule, and the preparation can be prepared as a powder ampoule in which the active ingredient is mixed with an inactive, water-soluble diluent. Parenteral administration can be intramuscularly, subcutaneously or intravenously. At the reported concentrations, the active substances did not damage tissues or vascular walls. The active ingredients may also be administered by infusion. Suppositories contain 2.20, preferably about 10 active ingredients, and are prepared from cocoa butter or synthetic waxes or fats suitable for this purpose (for example, Imhausen pulp manufactured in the Federal Republic of Germany).

27 6 5 9 9 0

The active ingredient content of cosmetic or skin-improving creams is 0.1-1 .mu.g / g. The ointment base may be hydrophilic or hydrophobic and contains conventional components: e.g. cholesterol, paraffin, glycerol, lanolin, cocoa butter, linseed oil, etc. The active ingredients may further be in the form of aerosols, the active ingredient content being also 0.1-1. / g. When formulating a perlingual tablet, this contains 10 ug of active ingredient, and the disintegration time of the tablet is 1/2 to 1 hour. Sustained release polymers can be prepared, for example, in the form of suspensions containing 1 to 5 .mu.g of active ingredient / g of polymer. Injectable preparations with a delayed action can be formulated either using high molecular weight polymers or salts of the compounds of the invention with high molecular weight organic bases (e.g. histone, prostamine), the ampoule containing 10-20 ug of active ingredient. Powders used for cosmetic or skin healing purposes are prepared with ordinary carriers (eg talc) and contain G, 1-1 μg active ingredient / g powder. In ophthalmology, compounds are formed from drops or ointment / miscible or immiscible ointments; the active ingredient content of these preparations is 0.1 to 1 ug / g. When administering the compounds of the invention to children, the dosage is about 0.3 μg per kg of body weight.

Sterile preparations are made by purely sterile filtration.

The desired preventive garmacological or cosmetic efficacy of the above formulations can be enhanced and supplemented with a variety of combinations. Among the possible biologically active additives, the following should be mentioned in particular, the use of which falls within the scope of the patent: vitamins A, C ~, E and K.it, trace elements, cortisone and its derivatives, progesterones, radiopharmaceuticals and immunosuppressive agents, antipsychotics , especially tranquilizers, thymoleptics, organosilicon compounds, gerantological preparations, blood cholesterol lowering agents, oral anti-diabetics, anti-inflammatory agents, antihistamines, etc. The dosage of these active ingredients is the same as the usual therapeutic dose of these agents alone.

28 6 5 9 9 0

The compounds of the invention may also be used as additives in pharmaceutical preparations and premixtures. In part, they have an effect on weight gain, in part they reduce the need for vitamin A or improve its metabolism. In addition, their use increases the resorption of trace elements whose blood mirror rises. When the compounds are used as a supplement to animal feed, their oral dosage is suitably 200 ng / kg per day. When mixed with feed, this corresponds to a concentration of about 1-2 yug / kg or 1-2 mg / ton, respectively, in the feed, i.e. 0.001-0.002 ppm concentration. Given this rather low concentration, its use as a feed additive is very economical. The compounds are preferably mixed with vitamin premixes, or used as microcapsules containing the compounds of the invention in addition to other necessary feed additives.

The compounds according to the invention can further be administered in the form of drinking water or an arrow or, optionally, also in the form of a spray.

In veterinary medicine, the compounds of the invention have similar uses as in human medicine, i. eg skin damage (flaking), wound healing, fractures, etc.

The invention is further illustrated by the following examples.

.o

Example 1; <1-L-glutamyltaurlin A. 40.85 g (0.11 moles) of carbobenzyloxy-L-glutamic acid β-benzyl ester (Liebigs Annalen 655 (J06-2) 200 are dissolved in 500 ml of acetonitrile. The solution is cooled to -1 5. While stirring, 15.4 ml (0.11 moles) of triethylamine are added dropwise to the solution, followed by 15.4 ml (0.11 moles) of chloroformic acid isobutyl ester, and the reaction mixture is stirred at -15 ° C. 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, and 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) in 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 water (250 ml) by shaking with the aqueous phase obtained with sodium carbonate solution. can be acidified with hydrochloric acid and shaken with ether to recover about 5 g of unreacted carbobenzyloxy-L-glutamic acid-oC-benzyl ester). The ethyl acetate phase is dried over anhydrous sodium sulfate and then evaporated to dryness in vacuo at 30 ° C to give a thick oily residue which soon crystallizes into a crystalline mass. This is triturated with absolute ether (250 ml), the crystals are filtered off. 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, N'-bis- (N-carbobenzyloxy - ") - (o <-benzyl) -L-glutamyl-cystamine are obtained, melting at 91-92 [deg.] C. Elemental analysis for C, N S (M = 859.05) 44 50 4 10 2

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 N, N'-bis-N-carbobenzyloxy- (N-benzyl) -L-glutamyl-4-cystamine a obtained in 1A 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 to the ice-cooled solution over 15 minutes. 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 11a. 28.5 g of carbobenzyloxy-N- (o (-benzyl) -L-glutamyl) litarin are obtained, and the crude product can be used without purification for the preparation of N, N-glutamyl taurine.

C. 26.32 g (55 mmol) of the carbobenzyloxy prepared in 1B are dissolved in 50 ml of glacial acetic acid. To the solution is added 4 moles of hydrogen bromine dissolved in 50 ml of glacial acetic acid. The reaction mixture is allowed to stand for 2 hours at room temperature, then evaporated to dryness in vacuo at 30 [deg.] C. The oily residue is dissolved in 170 ml of water and shaken with ether (5 x 70 ml). to dryness in vacuo at 35 [deg.] C. 20.42 g of β- ((<-benzyl) -L-glutamyltaurine are obtained, which is recrystallized from 90% ethanol, m.p.

30. Λ ^ ϋ5 990 R ^ (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. Dissolve γ- (C 1 -benzyl) -L-glutamyl taurine (20.42 g) obtained in 1 C above is dissolved in 150 ml of 1-n urea hydroxide solution. The solution is allowed to stand at room temperature for 4 hours, then transferred to a 2 x 100 cm column filled with Dowex 50 x 2 (Fluka, 100-200 mesh) in the H + phase. Elute with water. From the beginning of the elution, 300 ml of eluate are collected. This is evaporated to dryness in vacuo at 35 ° C, the oily residue is crystallized by adding 8-10 ml of water and about 100 ml of ethanol. Filter, wash with alcohol and dry to give 13.7 g of γ-L-glutamyl taurine. The crystalline product is recrystallized from 80% aqueous ethanol. 9.79 g of pure product are obtained, corresponding to a yield of 70% with respect to N, N'-bis-N-carbobenzyloxy-Y-benzyl) -L-glutanyl-cystamine.

Example 2: PL-glutamyl taurine A. 529 mg (1.1 mmol) of the carbobenzyloxy-β- (tX-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. at 4 hours. The solution is then shaken with ether (3 x 3 mL). The aqueous phase is transferred to a 1x20 cm column filled with Dowex 50 x 2 and eluted with water. Collect 50 ml of the solution and evaporate to dryness in vacuo at 35 ° C. Crude carbobenzyloxy-γ-L-glutamylurine is obtained, which is purified by paper electrophoresis at pH 6.5. The relative mobility compared to cysteic acid is 1.05.

(n-butano-1-pyridine-glacial acetic acid-water 15: 10: 3: 12) = 0.57.

B. The carbobenzyloxy-γ-L-glutamyl-taurine obtained in 2A 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 half a feel, then it is evaporated to dryness in vacuo at 35 ° C. The residue is triturated several times with ether and separated from it by decantation. The resulting 7-L-glutamyl taurine is recrystallized as described in Example 3.

31 65990

Example 3: N-L-glutamyl taurine 5.79 g (12.1 mmol) of the carbobenzyloxy-ΊΓ - (<X-benzyl) -L-glutamyl taurine prepared in Example 1B are dissolved in ethanol (100 ml) and water (25 ml). mixture and hydrogenated with shaking in the presence of 10% palladium on carbon 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 pentoxide. 3.1 g of N, L-glutamyl-aurine are obtained, which is very soluble in water and insoluble in alcohol. Addition of a small amount of water and alcohol in small portions causes the product to crystallize. The crystalline product melts at 202-204 ° C.

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-carbobenzyl: i.ox.-Y '- (<K-benzyl) -L-glutamyl-7-cystamine. The melting point of the pure product is 219-220 ° C.

r. -, 20 = +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.

Rj (n-butanol-pyridine-glacial acetic acid-water 15: 10: 3: 12) = 0.19

Analysis for the formula C_H N o, S (M = 254.29): '14 2 6

Calculated: C 33.07%, H 5.55%, N (1.02%, O 37.75%, S 12.61% Found: C 33.15%, H 5.76%, N 50.94 H, O 37.53%, S 12.17%.

In the IR spectrum, the characteristic groups have the following maxima:? (NH 4 and OH) 3600-2400; ylNH; amides) 3315; HCO; carboxyl) 1758; neo; amide) 1666; r (NH +) 1 576; VlSO ") 1296 and

J

Example 4: N-L-glutamyltaurine 5.42 g (11 mmol) of carbobenzyloxy-L-glutamic acid (Ύ-benzyl) -T-p-nitrophenyl ester (Chem. Ber. 96 (1963) 204) are dissolved in 50 ml of pyridine. The solution was cooled to 0 ° C and then added dropwise over a half hour 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 32 65990 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 carbobenzyloxy-1H- (o-benzyl) -L-glutamyltaurine triethylammonium salt are obtained. This substance is catalytically hydrogenated as described, for example, 3, and the solvent is removed by evaporation in vacuo. To remove triethylamine, the substance is dissolved in a small amount of water and the solution is taken up in a Dowex 50 x 2 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 5. 1.72 g of N, N-glutamyl taurine are obtained, corresponding to a yield of 68% with respect to taurine.

In the IR spectrum, the groups of cavacteria have the same maxima as in Example 3.

Example 5; L-glutamyltaurine A. 0.48 g (1 mmol) of carbobenzyloxy-O- (L-p-nitrophenyl ester) -glycine ethyl ester (Acta Chim. Sci. Hung.

65 (1970) 375 is dissolved in 6 ml of ethyl acetate. The solution is cooled with ice water. At 0 ° 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 for a while in ice water and then for 24 hours at room temperature, 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 carbobenzyloxy-o <-L-glutamyl- (1H-cysteamine) -glycine ethyl ester is obtained, corresponding to a yield of 85%.

Analysis for the formula C1QH 0.N-.S (M = 411.4):

I O 4D o J

Calculated: C 62.6 S, H 6.1%, S 7.8% Found: C 53.4%, H 6.5%, S 7.7%.

33 In the 65990 iR spectrum, the characteristic groups have the following maxima: V ~ (NH) 3310, VKC = 0; COOEt) 1748, "ΠθΟ; (Ζ)) 1690, Y '(C = O; amide) 1655 cm -1 · 100 mg of carbobenzyloxy-C 1 -L-glutamyl- (T-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-T-L-glutamyl-1- (T-taurine) -glycine ethyl ester (95 l) is obtained.

B. Dissolve 100 mg of carbobenzyloxy-cK-L-glutamyl (Τ'-taurine) -glycine ethyl ester prepared in 5A 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 a sealed bomb tube. 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 a positive ninhydrin spot. The product is δ (-L-glutamyl- (Τ'-taurine) -glycine, yield 0.06 g (88%).

Analysis for Coh1-, N_0tS (M = 311.3) y I / 3 /

Calculated: S 10.3% Found: S 10.0%.

In the IR spectrum, the character groups have the following maxima: V ~ (NH +) 3100 (Broad), ΥΊΟΗ; COOH) 3200 (broad), 1T (C = O; C00H) 1730, r (C = O; amide I) 1680, Y1C = O; amide-II) 1560, HS = 0; SO OH) 1220 ( intens- _ - 1 ^ winged),) 1045 (intense) cm

Hydrolysis: 20 mg of substance 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.

C. Dissolve 100 mg of O (-L-glutamyl- (β-taurine) -glycine prepared in 5 B in 25 N of 0.2 M ammonium bicarbonate buffer at pH 8.5. Add 1 mg of carboxypeptidase A to the solution: dissolved in 0.5 ml of water (Enzyme manufacturer: Serva, Heidelberg) The mixture is kept for 24 hours in a thermostat at 37 [deg.] C., then lyophilized, and the dry lyophilisate can be detected in 34 6 5 9 9 0 YL-glycamyl laurine and glystinl. Pure YL-glutamyl-taurine is obtained by electrophoresis or chromatography on a Dowex 50 column.

Example 6: f'-L-glutamyltaurine A. 0.47 g (1 mmol) of carbobenzyloxy-O (-L-glutamyl- (γ * -p-nitro-phenyl ester) -glycine methyl ester are dissolved in 6 ml of pyridine.

To an ice-cooled solution is added 0.125 g (1 mmol) of taurine in 2 ml of water, then 0.28 ml (2 mmol) of triethylamine in such small portions that the solution remains clear at all times. The reaction mixture is allowed to stand at room temperature for 3 days. After evaporation to dryness in vacuo, an oil is obtained which is thoroughly digested in ether, then in petroleum ether and finally dried in vacuo over sulfuric acid. Carbobenzyloxy-L-glutamyl- (γ'-taurine) -glycine methyl ester is obtained.

B. 100 mg of the carbobenzyl ± oxy-O <-L-glutamyl- (1H-taurine) -glycine methyl ester prepared in 6A above is reacted at room temperature with 2N hydrochloric acid in 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 24 hours. The resulting precipitate 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 hydrogen bromide of the β-L-glutamyl-γ "-taurine) -glycine methyl ester is practically pure.

C. The salt obtained in 6B 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 ion exchanged at 10 r in Dowex 50 H form, 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 N - (- L-glutamyl- (Y'-taurine) -glycine are obtained. In the IR spectrum, the caviter groups have the following maxima: Γ (ΝΗ) 3310, jr (ΝΚί) 3100 (broad), V (C = O; C00H) 1730, V (C = O; amide I) 1650, Y * (C = O; -1 amide II) 1570, Y (S = O) 1220 (intense), 1045 (intense) cm .

(3i 65990 D. Dissolve 100 mg of Crt-L-glutamyl- (T-taurine) -glycine prepared in 5B in 25 ml of 0.2 M ammonium hydrogen carbonate buffer, pH 8.5. To the solution is added 1 mg of carboxypeptidase. A. dissolved in 0.5 .mu.l of water (Enzyme manufacturer: Serva, Heidelberg) The mixture is kept for 24 hours in a thermostat at 37 [deg.] C., then lyophilized, and the dry lyophilisate can be detected in N, L-glutamyltaurine. and glycine Pure 'Y'-L-glutamyl taurine is obtained by electrophoresis or chromatography on a Dowex 50 column.

Example 7: β = -L-glutamyl skin motaurine A. 1.083 g (2.2 mmol) of carbobenzyloxy-L-glutamic acid (<κ-benzyl) -γ-nitrophenyl ester are dissolved in 6 ml of a 2: 1 aqueous mixture of pyridine. . To the solution is added first 278 mg (2 mmol) of homotaurine, then 0.59 L (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 p-nitrophenol in a continuous extractor with ether for 8 hours. The aqueous phase is evaporated in vacuo. 1.68 g of carbobenzyloxy-Y- (δ-benzyl) -L-glutamyl] homotaurine are obtained.

B. The material prepared in 7A above (1.68 g) is dissolved in 10 ml of 50% aqueous ethanol, 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 transferred to a Dowex 50 X 2 column (1 x 35 cm) in H + form to remove trethylamine. Elute with water. The eluate is taken up in 50 ml and then evaporated to dryness. The residue contains 440 ml of γ-1-glutamylhma-taurine, corresponding to a yield of 82%. Electrophoresis at pH 6.5 shows 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 36 6 5 9 9 0 In the IR spectrum, the characteristic groups have the following maxima: V (-NH) 3345; Y (NH 4, OH) 3200-2200; Γ (ΟΟ) 1740; Y (CO-NH), 1645; V'tSO ") 1240, 1190, 1150 and 1038; ^ (- NH) 1 570 and 1 535; (SO3) 590 cm-1.

Example 8: <PL-Glutanyl-N-methyltaurine A. 1.083 g (2.2 mmol) of carbobenzyloxy-L-glutamic acid ((X-benzyl) -β-nitrophenyl ester are reacted with N-methyltaurine (278 mg, 2 mmol) as described in Example 7A, 1.59 g of carbobenzyloxy- (OS-benzyl) -L-glutamyl-N-methyltaurine are obtained.

B. The entire amount of the material prepared in 8A above (1.59 g) is catalytically hydrogenated as described in Example 7B. 423 'of Y-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 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; «Trans-L-Glutamyl-L-cysteic acid A. 2.87 g (6.6 mmol) of carbobenzyloxy-L-glutamic acid (N-benzyl) -N-nitrophenyl ester are dissolved in 20 ml of pyridine and 1.25 g are added. g (6 mmol) of L-cysteic acid monohydrate dissolved in a mixture of water (17 ml) and pyridine (17 ml). An additional 2.6 mL (18.6 mmol) of triethylamine is added, after which the reaction mixture is allowed to stand at room temperature for 72 hours. The solution is then evaporated in vacuo at 30 ° C. The residue is dissolved in 20 ml of water, the solution is acidified with concentrated hydrochloric acid and then shaken with ether (15 x 10 ml). The aqueous phase is evaporated in vacuo at 35 ° C. Carbobenzyloxy-? "- (1'-benzyl) -L-glutamyl-L-cysteic acid is obtained.

B. Dissolve the product prepared in 9A above in 20 ml of water, add 0.3 g of 10% palladium-on-carbon, and pass hydrogen through the suspension for 3 hours. The reaction mixture is worked up as described in Example 7B. T-L-glutamyl-L-cysteic acid is obtained, melting at 187 ° C. By paper chromatography, the relative mobility to cysteic acid is 1.21 at pH 6.5 and 0.54 at pH 1.8.

*

37 6 5 9 9 O

In the IR spectrum, the characteristic groups have the following maxima: ΎΊΝΗ; amide:) 3423; Y (NH 4, OH) 3300-2200; Y ^ CO; carboxyl) 1748 and 1712; V "(C0; amide) 1627; Y" (NH; amide) 1 527; 77503) ^ 21, 1109 and 104 3; <MSO ^) 525 cm ^.

Analysis for C 9 H 18 N 2 O 2 S (M-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 S.

Example 10: β-L-glutamylcholamyl phosphate; A. 1.083 g (2.2 mmol) of carbobenzyloxy-L-glutamic acid - (<X-benzyl) -T-p-nitrophenyl ester are dissolved in 6 ml of a 2: 1 mixture of pyridine and water. To the solution is first added 282 mg (2 mmol) of cholamine 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-Y - ((X-benzyl) -L-glutamylhomotaurine (Example 7A) to give 1.25 g of carbobenzyloxy-TT - (- Benzyl) -glutamylcholamine phosphate.

B. The entire amount of product obtained in 10A above (1.25 g) is hydrogenated from the catalytic deprotector. Hydrogenation and ion exchange purification are performed as described for the preparation of γ-L-glutamyl homotaurine (Example 7B). 470 mg of 7-L-glutamyl.cyclaminiphosphate are obtained, corresponding to a yield of 91%. However, as can be seen in paper electrophoresis, the product contains about 15-20% cholamine phosphate as an impurity. Electrophoresis, among others, can be used as the purification method.

In electrophoresis, the substance migrates at both pH 6.5 and pH 1.8 per anode. The relative mobility to cysteic acid is 0.15 at pH 6.5 and 0.36 at pH 1.8.

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

Example 11: N-L-glutamyl homotaurine A. 526 mg (1.1 mmol) of carbobenzyloxy-L-aspartic acid (6X-benzyl) -β-nitrophenyl ester (Chem. Ber. 97 (1 964) 1709 are dissolved in 5 ml of: The solution is cooled to 0 [deg.] 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. 48 hours, then evaporated in vacuo, the residue is dissolved in 5 ml of water 65990 38 and initially 1N hydrochloric acid is added to the yellow solution until the color disappears, the solution is shaken with ether (10 x 5 ml) to remove p-nitrophenol and the aqueous phase is evaporated in vacuo.

4 78 mg of carbobenzyloxy-A - (- benzyl) -1-aspartyl-taurine are obtained.

B. Dissolve the entire amount of the product obtained in 11A above in 6 ml of 50% aqueous ethanol. 100 mg of 10% palladium-on-carbon are added to the solution, and the mixture is then boiled for 4 hours by passing hydrogen gas through it. After filtration and evaporation in vacuo, triethylamine is removed from the product by the ion exchange method described in connection with the preparation of N, L-glutamyl homo-taurine (Example 7B). 172 mg / 3-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: β-L-aspartyl homotaurine A. 526 mg (1.1 mmol) of carbobenzyloxy-L-aspartic acid (CK-benzyl) -β-nitrophenyl ester and 139 g (1 mmol) of homotaurine are reacted as described in Example 11A. way.

Carbobenzyloxy-N - ((X-benzyl) -L-aspartyl taurine is obtained.

B. The product from 12A above is catalytically hydrogenated as described in Example 7B. 203 mg (84 £) of N-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.

R f (n-butanol-pyridine-glacial acetic acid-water) 15: 10: 3: 12) = 0.17. Example 13: η 5 -L-aspartylcholamine phosphate A. Carbobenzyloxy-L-aspartic acid (β-benzyl) - β-p-nitrophenyl ester and cholamine phosphate are reacted with each other as described in Example 10A. Carbobenzyloxy, β - (<X-benzyl) -L-aspartylcholamine phosphate is obtained.

B. The material from 13A above is catalytically hydrogenated as described in Example 7B. Β-L-aspartylcholine phosphate is obtained.

39 65990

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 0.81 at pH 6.5 and 0.40 at pH 1.8.

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

Example 14: Monosodium salt of β-L-glutamyl taurine 25.4 mg (0.1 mmol) of T-L-glutamyvlitaurine are dissolved in 2 ml of water, and 10 ml of 0.01 N sodium hydroxide solution is added to the solution. The mixture is evaporated to dryness in vacuo at 30 ° C. The white crystalline residue is dried in a desiccator over phosphorus pentoxide.

The monosodium salt of Y '-'-L-glutamyl.taurine is obtained. The product does not o ...

there is no sharp melting point, it begins to shrink into the pile at 200 C and the color becomes constantly darker, and at about 250 ° C it charts.

The product is equally poorly soluble in methanol and ethanol.

O

Example 15: -g-L-glutamyl taurine r-L-glutamine taurine (prepared according to Examples 1-6) is purified by recrystallization as follows: 300 mg of the crude substance are dissolved in 5 ml of anhydrous dimethyl sulfoxide at room temperature with stirring. The opalescent solution is filtered and the filter is washed with 0.5 ml of dimethyl sulfoxide. The filtrate and washings are combined and 55 ml of absolute ethanol are added.

The mixture is allowed to stand at room temperature for 12 hours, then the substance is filtered, washed with 2.5 ml of absolute ethanol and dried in a vacuum desiccator over phosphorus pentoxide until a constant weight is reached. 240 mg of crystalline β-L-glutargyl taurine are obtained (yield in recrystallization: 80%).

Instead of ethanol, the same amount of dioxane, ether or acetone can also be used for recrystallization. The quality of the crystalline product is the same in all cases. Melting point (according to Boetius): 218-219 C. The product is uniform by thin layer chromatography.

Example 16: N, N-Glutamylcholamine phosphate

Carbobenzyloxy-γ- (α-benzyl) -L-glutamylcholamine is prepared in a manner suitable for the preparation of glutamic acid N-amides (Acta Chim.Acd.Hung. 64 (1970) 285. 4.14 g of this product (10 mmol) are dissolved in 50 ml. 9 g (33 mmol) of diphenylphosphoric acid chloride are added to the solution, the reaction mixture is kept at 0 [deg.] C. for 12 hours, then diluted with 80 ml of 1140 6 5 990 chloroform, the separated product is filtered off, washed with dilute hydrochloric acid and then dried with water and finally dried in a desiccator over potassium hydroxide The dry substance is dissolved in 15 ml of glacial acetic acid containing 3.3 mol / l of hydrogen bromide and the solution is allowed to stand for 15 minutes, then evaporated to dryness in vacuo at 35 ° C. The residue is dissolved in 30 ml. sodium hydroxide solution and the solution is allowed to stand at room temperature for 1 hour, then the pH is adjusted to 4 and the solution is extracted with ether (3 x 30 ml) to remove phenol and benzyl alcohol. Dowex 50 into a statue and eluted with water. The eluate is evaporated in vacuo and the residue is crystallized from 2: 1 acetone-water. 0.8 g of r-L-glutamylcholamine phosphate is obtained.

Example 17: N-L-glutamylcholamine phosphate 4.68 ml (5 mmol) of phosphorus oxychloride are added dropwise to 1.8 ml of water under cooling and stirring (Biochem. Preparations 6 (135c) 76). To the solution is added in small portions 1.9 g (10 mmol) of T'-L-glutamylcholamine (prepared from carbobenzyloxy-Y- (O <-benzyl) -L-glutamylcholamine according to Example 16). The mixture is stirred at 60 ° C for 2 hours, after cooling, 0.72 ml of water is added thereto with stirring, and then it is allowed to stand at room temperature for 2 hours. 10 ml of 96% ethyl alcohol are then added dropwise with stirring, followed by 10 ml of ether. The reaction mixture is quenched at 4 ° C overnight, then 5 ml of 96% ethanol are added. The precipitated substance is filtered off, washed first with ethanol, then with ether and finally recrystallized from a mixture of ethanol and water. 1.75 g of r-L-glutamylcholine phosphate are obtained.

Example 18 N-L-glutamyl itaurin 4.14 g (10 mmol) of carbobenzyloxy-γ- (o-benzyl) -L-glutamylcholamine are dissolved in 40 ml of pyridine and the solution is cooled to -10 ° C. 2.1 g (11 mmol) of p-toluenesulfonyl chloride are added in small portions with vigorous stirring. The reaction mixture is stirred at 0 ° C for 3 hours, then poured onto 40 g of ice. The separated precipitate is filtered off, washed with water and finally crystallized from a mixture of ethanol and petroleum ether. The product is catalytically hydrogenated as described in Example 3. The dried product obtained after hydrogenation is dissolved in 30 ml of water, 10.1 g of 41,69900 (40 mmol) of sodium sulfite heptahydrate are added to the solution. The solution is stirred at 40 ° C for 24 hours and then evaporated to dryness in vacuo. The residue is dissolved in a small amount of water and taken up in a Dowex 50 column from which the product is eluted with water. The eluate is evaporated in vacuo and the residue is dried over potassium hydroxide. Crystallization from 80% ethanol gives 1.6'-L-glutamyltaurine.

Example 19; fr-L-glutamyltaurine To 4.14 g (10 mmol) of carbobenzyloxy-Υ- (O-benzyl) -L-glutamylcholamine is added 15 ml of thionyl bromide, and the mixture is stirred for 3 hours. Ether is then added and the separated precipitate is filtered off and recrystallized from a mixture of acetone and petroleum ether. The resulting substance is dissolved in a mixture of dimethylformamide and water, and 10.1 g (40 mmol) of sodium sulfite heptahydrate are introduced into the solution with stirring. The mixture is stirred at room temperature for 24 hours, then for a further 6 hours at 50 ° C and finally filtered. The clear solution is evaporated in vacuo and the residue is hydrogenated from the catalyst as described in Example 3. The dried substance obtained after hydrogenation is dissolved in a small amount of water, and the solution is taken up in a Dowex 50 column and eluted with water. The eluate is evaporated in vacuo and the residue is crystallized from 80% ethanol. 1.2 g of N, N-glutamyvlitaurine are obtained.

Example 20: N-L-glutamyltaurine 1.71 g (10 mmol) of carbobenzyloxy-L-glutamic acid O-benzyl ester are dissolved in 60 ml of acetonitrile, the solution is cooled to -15 ° C and 1.4 g are added with stirring. ml (10 mmol) of chloroformic acid isobutyl ester, then 1.4 ml (10 mmol) of triethylamine The reaction mixture is stirred at -15 [deg.] C. for 30 minutes 30 minutes, then 2.05 g (10 mmol) of bromoethylamine hydrobromide, 1.4 ml (10 mmol) of ) triethylamine and 40 ml of acetonitrile are cooled to -15 [deg.] C. The mixture is stirred at 15 [deg.] C. for 2 hours, then at room temperature for a further 4 hours.

It is then filtered and the filtrate is evaporated in vacuo at 35 ° C. The residue is dissolved in a mixture of dimethylformamide and water, and 10.1 g of sodium sulfite heptahydrate are added to the solution. Further work-up of the reaction mixture is carried out as described in Example 19 to give 1.45 g of Y-L-glutamylurine.

Example 21; N-L-glutamyltyrine 3.02 g (10 mmol) of the sodium salt of carbobenzyloxy-L-glutamine (Liebigs Annalen 640 (1961) 145) are dissolved in 50 ml of dimethylformamide. An oil suspension containing 12 mmol of sodium hydride is added to the solution, then the solution is heated 2 A solution of 2.11 g (10 mmol) of sodium bromoethanesulphonate in 50 ml of dimethylformamide is then added dropwise to the mixture and the mixture is heated for a further 2 hours, then evaporated to dryness in vacuo, the residue extracted with ether and dried. , the aqueous solution is taken up in a Doewx 50 column and eluted with water. The eluate is evaporated to dryness in vacuo and the residue is dried over potassium hydroxide. The dry substance is catalytically hydrogenated as described in Example 5. Recrystallization from ethanol / water gives 1.55 g of "-L-Glu-tamyltaurine.

Example 22: β-Glutamylautin 2.58 g (10 mmol) of L-N- (2,6-dioxo-3-piperidyl) -phthalimide (J.Pharm.Sci. 57 (1, 68) 757) dissolved in sodium ethanol solution prepared in absolute ethanol. The solution is evaporated in vacuo, the residue is dissolved in 50 ml of dimethylformamide and 2.25 g (11 mmol) of sodium bromomethanesulphonate are added to the solution with stirring. The mixture is heated for 2 hours, then evaporated in vacuo. The residue is dissolved in water and the solution is taken up in a Dowex 50 column and eluted with water. The eluate is evaporated in vacuo and the residue obtained is boiled in 100 ml of 0.5 N hydrochloric acid for 3 hours. After evaporation of the solution, 30 ml of ethanol and 0.7 ml of 72% hydrazine hydrate are added to the resulting substance, and the mixture is boiled for 3 hours. The mixture is then evaporated in vacuo. The residue is dissolved in 25 ml of 2N hydrochloric acid, the mixture is kept at 50 ° C for 10 minutes and then for 30 minutes at room temperature. The separated solid is removed by filtration, the filtrate is evaporated in vacuo and the residue is dried in a desiccator over potassium hydroxide.

The dry substance is dissolved in a 90: 8: 2 mixture of water, acetic acid and formic acid and the solution is taken up in a Dowex 1 column (2 x 100 cm). The statue was pre-equilibrated with said solvent mixture. First elute with said solvent mixture, recover 400 ml of eluate. This fraction contains β 2 -L-glutamyl taurine. It is then eluted with 0.5 N hydrochloric acid and 400 ml of eluate are again recovered. This fraction is evaporated at 35 ° C in vacuo. The residue is dried in a desiccator over solid potassium hydroxide and the product obtained is crystallized from 80% ethanol. 1.07 g of T'-L-glutarayl taurine are obtained.

Example 23; Β-L-glutamyl taurine 4.43 g (10 mmol) of the dicyclohexylamine salt of carbobenzyloxy-L-pyroglutamic acid (Liebigs Annalen 640 (1961) 145), (10 mmol) of taurine and 0.84 g (10 mmol) of sodium hydrogen carbonate are dissolved in 50 ml of water. . The solution is heated for 4 hours (or allowed to stand at room temperature for 24 hours) and then the solution is evaporated in vacuo. The residue is dissolved in water and the solution is taken up in a Dowex 50 column and eluted with water. The eluate is evaporated to dryness. The resulting substance is catalytically hydrogenated as described in Example 3. After crystallization from the ethanol-water mixture, 2.03 g of N-L-glutamyl taurine are obtained.

Example 24: N-glutamylcholamine sulphate 7.38 g (15 mmol) of carbobenzyloxy-L-glutamic acid (N-benzyl) -β-p-nitrophenyl ester and 1.41 g (10 mmol) of collarine sulphate (see PA Leighton, J. Am. Chem. Soc., 69 (1947) 1540)) is reacted with each other as described in Example 7A. Work-up of the reaction mixture, deprotection by catalytic hydrogenation and purification of the product are carried out as described in Example 3. 1.75 g of N '- L-glutamylcholamine sulphate (64.7%) are obtained, m.p. 164-166 ° C.

In the IR spectrum, the characteristic groups have the following maxima: V (-NH) 3365; T "(NH 3 + OH) 3200-2400; T '(CO) 1748; Y | cO-NH) 1660; (SO, -) 1260, 1186, 1155, 989 and 829; <ζ (-ΝΗ-) 1565 and 1540, S (SO ~) 580 and 595 cm -1.

Claims (5)

1. R -HN-CH-CO-A «jVn CO-N-CH- (CH2) t ~ B2 (V) 1 h 5. 1 wherein R, R, n 7a t are as defined above, A is (1) hydroxy, (2) C 1-4 alkoxy, (3) C 1-8 aralkoxy optionally substituted in the p-position by alkoxy or nitro, or (4) a group of formula (b) wherein R 6, Y and r have the same meaning as above, R is (1) hydrogen, (2) C 1-4 alkoxycarbonyl or 46 65990 (3) C 1-8 aralkoxycarbonyl optionally substituted by one or more halogen towers, or by one or more 2 alkoxy or nitro groups, 3 is -SH, -SO 4, -OH, p-toluenesulfonyloxy, halogen, or -SO, R 1, where R 1 * is C 1-4 alkoxy or aralkoxy, is oxidized or, if the compound of formula (V) contains a sulfonic acid ester group, partial hydrolysis is carried out, or 2 if B is p-toluenesulfonyloxy or halogen , reacting a compound of formula (V) with an alkali sulphite or alkali hydrogen sulphite to give compounds of general formula (I) in which B1 is -SO 2 OH, or if the compound of formula (V) contains hydroxyl groups, it is esterified with sulfuric acid or phosphoric acid or a derivative thereof to give compounds of general formula (I) containing a hydrogen sulphate or dihydrogen phosphate group and, if desired, cleaving protecting groups attached to or bound to the β-amino and / or t'-carboxyl groups, t ai e) for the preparation of compounds of general formula (1) in which B2 3 is a group containing a sulfonyl, a compound of general formula (VI) R 4 5 -HN-CH-CO-A1 (CH) i 2 n 12 (VI ) CO-NR tVi '1 1 3 R 11 12 wherein R, A and n are as defined above, R is hydrogen or 3 C 1-6 alkyl and R is an alkali metal, is reacted with an alkali metal salt of a haloalkylsulphonic acid, followed by optionally cleaving protecting groups attached to or bound to the β-amino and / or β-carboxyl groups, or 2 f) for the preparation of compounds of general formula (I), wherein B is -SO 2 OH, a compound of general formula (VII) 4 <WcH- (CH2) t-S- (VII) 5 RR 65990 47 4 wherein A is hydroxyl, aralkoxy or preferably benzyloxy, substituted aralkoxy, preferably p-methoxybenzyloxy or 5 7 p-nitrobenzyloxy and R, R, R, n and t have the same meaning than 7 4 above is oxidized, after which the protecting groups R and A are cleaved, or g) according to the general formula (I) for the preparation of compounds wherein 1 is -SO 2 OH, a compound of general formula (XXII) -C? NMR (xxii) 0 wherein n is an integer of 1 or 2 and M is sodium or potassium is reacted with a compound of general formula (ΧΧΙΙΙ) Hal - CH - (CH „) - SO„ OH (ΧΧΙΙΙ ) 1. ztz R 5 wherein R and t are as defined in formula (I) and Hal is halogen, or a salt thereof, a compound of formula (XV) 0 O Il !! , r \ \ v i | N-CH N-CH- (CHO) -SO., OH (XV) k / '-?' IKO 2'n O 1 wherein R, n and t are as defined above is partially hydrolysed with a dilute acid or base to give a compound of formula (XXIV) 0 65990 48 UN - (^ H-COOH "" / 'iVn Ö CO-NH-CH- (CH 2) -SO-OH (XXIV) 15. i R 5 wherein R, n and t are as defined above are reacted with hydrazine, or h) a compound of general formula (XVIII) R7-NH-CH-CO-A4 (CH0) (XVIII) I 2? 4. CCHA 7 wherein A, R and n are as defined above and A is hydroxyl, p-nitrophenyloxy, pentachlorophenyloxy or C 2-6 alkoxycarbonyloxy, is reacted with a compound of general formula (XX) HijJ- <j : H- (CH2) t-B1 (XX) R R5 15 wherein B, R, R and t have the same meanings as above, after which the resulting compound is cleaved with protecting groups A and R, preferably the protecting group R7 is cleaved by acidolysis and protection - Group A is cleaved by alkaline hydrolysis, which cleavages are carried out in an arbitrary order, and if desired the resulting compound is converted into a pharmacologically acceptable salt or liberated from the salt and / or the compound is isolated as its optically active isomers using an optically active starting material or racemate resolution. 65990 49 For the preparation of a therapeutically active amino acid derivative with all the compounds of formula (I) h2n-ch-co-oh «PV n, (I) CO-N-CH- (CH2) -B 5 RR 5 1 color R is derived from C 1-4 alkyl, R is derived from carboxyl / or B is -SC 2 OH, -OSO 2 OH or, -OPO (OH) 2, or 1 or 2 or 2 and h or 1 or 2 of the same pharmacologically active and optically active isomeric compounds are shown in Figure 7 a) in the group R of the genomic acidolysis or hydrolysis of the compounds of formula (II) R7-HN-CH-CO -OH (CH) i 2 n. CO-N-CH- (CH ")" B (II) kk5 7 color R is (1) -? - alkoxycarbonyl or (2) C7-9 ~ aralkoxycarbonyl, optionally substituted with a halogen atom S1 or a halogen atom alkoxy- or nitro groups, and R, R ~, B, n and t in any of the following conditions, or b) in the case of all the compounds of formula (III) H „N-CH-CO-A1 1 i {CH-J) I 2 n ^ CO-N-CH- (CH) -B1 (III) II 5 1 r RR color A är (1) C 1-4 alkoxy, (2) C 1-4 aralkoxy, optionally with alkoxy or nitro substituents i p -stellation, or (3) and groups of formula (b) 50 65990 (-NH-p-CO) rY (b) color R is an alkyl or C 1-4 alkyl, Y is hydroxyl or C 1-4 alkoxy 1-4 51 1-4 och r är ett helt tai 1-10 och R, R, B, n och t betecknar decamina som ovan, undercast alkaline or enzymatic hydrolysis, en acidolys eller hydrogenolys, eller 7 3 c) skyddsgrupperna R och A veltidigt avlägsnas genomic acidolysis, alkaline hydrolysis or hydrogenolysis with the addition of the formula (IV) R7 -HN-CH-CO-A3 (CH) | 2 π ^ CO-N-CH- (CH.J ^ -B '(IV) II 5 1 - RR 3 color A är (1) C 1-4 alkoxy or (2) C 1-6 aralkoxy, optionally alkoxy or nitro substituents on the p-ring, and R, R5, R7, B, and octyl of the decamate as defined above, or d) in the case of the lower form (V) R1-HN-CH-CO-A1 (CH ) I 2 n 2 CO-N-CH- (CH) -B (V) fc R5 color A is (1) hydroxy, (2) C1-4 alkoxy, (3) -galalkoxy, optionally with alkoxy or nitro substituents i p-ställningen, eller (4) en grupp med formuleln (b), vari R6, Y and R betecknar decamma som ovan, R är (1) väte, (2) C ^ _4 ~ alkoxycarbonyl eller (3) C ^ _g -aralkoxycarbonyl, optionally substituted with a halogen atom or an alkoxy alkoxy or 2 nitro group, B is -SH, -SO 4 -, -OH, p-toluenesulfonyloxy, halogen or -SO 2 R 1, or R 1 <1 or C _4-alkoxy or aralkoxy oxides or compounds of formula (V) in which the sulfonyl ester group is used, partially hydrolysed, or 65990 2 of B and p-toluenesulfonyl x or halogen, the compounds of formula (V) with alkali sulphite or alkali bisulphite, are different from the compounds of formula (I), with B and -SO 2 OH, or with compounds of formula (I) having a hydroxyl group, for which they are in the case of phosphorus, or in the case of derivatives of the same formula (I), in the case of a bisulfate or a dihydrogen phosphate, , or (e) in the case of a compound having the formula 1 (I), a group B in which the sulfonyl is used in the formula (VI) R1-HN- ^ H-CO-A1 (CH2) (VI) in ^ λ όθ-Ν-R k1 2 11 12 color R, A och n betecknar decamma som ovan, R är väte eller 1 3 C 1-4 -alkyl och R is en alkali metal, omsättes med ett alkalimetall-salt av en halogenalkylsulfonsyra, varpä possibly with the addition of 1-amino- and / or carboxyl groups and a fusion group, or f) for a compound of the formula 1 (I), wherein B is -SC 2 OH, and for the formula (VII) R 3 -HN-CH-CO-A4 and CH )! 2 n CO-N-CH- (CH „) -S- (VII) II s 2 t RR 2 7 2 cyclo and oct, R, R, n and t in the case of a single ring, oxide, varp 3 3 avlägsnas skyddsgrupperna R and A; or 4 with A hydroxyl, aralkoxy or benzyloxy, substituted with aralkoxy, p-methoxybenzyl or p-nitrobenzyl 52 65990 g) for the conversion of the formulation with all of the compounds of formula (I), in the case of the formula (XXII) O o ^ 1 j yi \ s ^ c \ (xxii) I N - CH NM I! Ij <CH2) nC, o color is not helt or 1 or 2 or M is sodium or potassium, in addition to the formulation of all formulas (XXIII) Hal-CH- (CHJ -SCUOH (ΧΧΙΙΙ) k5 2 1 'color och t betecnar decamma som i formeln (I) och Hai är halogen, eller med ett sait därav, den erhällna föreningen med formuleln (XV) 0 O Ϊ ^ 'x. (xv) N-CH N-CH- (CH2) -SO_OH Ok: / | | i5 2 1 2 Ö (° η2) ^> 0 e, color R ~, n och t betecknar decamma som ovan, hydrolyser batches with enveloped syrups or bases and underlying compounds with a formulation of 0 I V- £ H-COOH ( XXIV) c '(fH2> n J CO-NII-CH- (CH2) t ~ S020H I5 5 color r, n och t betecknar decamma som ovan, omsättes med hydazm, eller