DK159267B - METHOD OF ANALOGUE FOR THE PREPARATION OF TAURIN DERIVATIVES OR HOMOTAURIN DERIVATIVES OR PHYSIOLOGICALLY ACCEPTABLE SALTS THEREOF - Google Patents

Description

DK 159267 B

The present invention relates to an analogous process for the preparation of novel taurine derivatives or homotaurine derivatives having valuable biological and pharmaceutical effects.

The 5 compounds prepared by the process of the invention have the general formula R1-NH-CH-COA

(CH,) n I

I i n CO-N- (CHO) .- SO, H i / ti

10 R

wherein 1 1 A, R, R, n and t have the meanings set forth in the preamble of the claim or are physiologically acceptable salts or optically active antipodes thereof.

The compounds of the present invention have valuable biological or pharmacological effects. All the compounds of the invention are novel.

Among the compounds of the invention, due to its biological action, particular emphasis is given to γ-L-20 glutamyltaurine corresponding to the formula

H0 N-CH-COOH

2 i

? H

I ά XXIV

CH0 25 I 2 co-nh-ch2-ch2-so2oh and having a broad therapeutic and preventive spectrum of action against morbid changes there.

30 can be traced back to damage by "AGAS" (the aerobiospheric genetic adaptation system). #

To illustrate the concept of "AGAS", the following are the main tissues and organs that form this system.

(a) all biological interfaces in contact with the external air such as the biosphere (skin and skin formation, eye cornea and conjunctiva, oral and pharyngeal cavities, respiratory tract and lung); b) skeleton and joints (tubular and spongy bones, ball joints, synovial membranes, skeletal muscle); 2

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(c) the organs involved in the regulation of the ion household (transepithelial transport system: intestinal tracts and renal duct); (d) the tooth kit 5 needed for comminution of the nourishment 5 (rooted in the tooth vaults); (e) hearing, smell and voice organs.

Thus, the compounds of this invention exert a favorable biological or therapeutic effect on the organs or tissues of the AGAS system enumerated herein.

In addition, the compounds prepared by the process according to the invention act on the following functions which are related to the AGAS system: radiation protection, favoring wound healing, general activating effect on mesenchymal, protection against the ever-increasing infection and contamination danger of skin and mucous membranes. (Lysozyme production of the moist mucosa, activation of the gut epithelium in the respiratory tract, etc.), increased protection against the infections caused by viruses and fungi.

The compounds of the present invention are effective against the ever-increasing 20 stress effects associated with mainland life (e.g., meteorological influence, large differences between day and night temperature, increased risk of injury) as they stabilize. adaptation syndrome and at the same time avert the peripheral tissue damage of the glucocorticoids (such as damage to the connective tissue, injuries to 25 bone matrix components, etc.). Development of immune homeostasis (increasing body recognition of which cells are suitable and which are not).

The compounds prepared by the process of the invention exert their effect, both immediately and partly above the regulation of vitamin A metabolism, in the production of vitamin A metabolites of a stronger polar nature. This effect is comparable to the effect of the parathormone on the 25-hydroxycholecalciferol-Ια-hydroxylase enzyme in the renal tract. The directions of action of the compounds are as follows: 35 A) Vitamin A grade effects: a) Pharmacological and biochemical effects:

DK 159267B

3 increasing effect on chondroitin sulfate synthesis; beneficial effect on the wound healing or on the experimentally impaired wound healing in rats and dogs by cortisone administration; potentiating effect on vitamin A's effect in rats and chickens in 5 experimentally induced hypo- or hypervitaminoses; attenuating effects on the ulceration-related stress effects in rats; beneficial effect on the degranulation of mastocytes; increasing effect on the production of lysozyme; effect on household traceability (silicon, zinc, copper, manganese, fluorine); emerging effect on epithelial formation; promoting effect on the alkaline phosphatase activity; the effect on the granuloma bag formation (granuloma sac formation) induced by local action of vitamin A; the extremely flat course of the dose-effect curve or the change of effect sign at large doses; activating effect on the Golgi apparatus; beneficial effect on the formation of mucus or goblet cells; increasing effect on the concentration of vitamin A.

b) Clinical-therapeutic effects: keratoconjunctivis sicca; Sjogren's syndrome; rhino-laryngo-pharingitis sicca; ozæna; 20 chronic bronchitis; sinobronchitis; mucoviscidosis; constitutional lung disease in young children; periodontal disease; skin and mucosal susceptibility to viruses and fungi; cortisone antagonistic effect; beneficial effect on healing of operative and mucosal wounds; erosio colli; pruritus-like disorders; reduction of 25 sense of smell and taste.

B) Vitamin A-grade effects (a) Pharmacological and biochemical effects: effects on the blood sugar level of a transient decrease; increasing effect on phosphaturia, lowering effect on serum phosphate level; radiation protective effect; reducing the time required to reach the goal of maze trials in inactivated animals; reducing effect on experimentally induced fluorine and cadmium toxicosis; increasing effect on the cyclic adenosine monophosphate depletion of the kidneys; attenuating effect on the symptoms of experimentally induced lathyrism; decrease in histamine sensitivity; enhancing effect on liver enzyme tyrosine aminotransferase activity.

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b) Therapeutic effects: weak radiation damage; vitiligo; muscle hypotonia; psychoenergetic effect; beneficial effect on involutional and gerontological conditions as well as on the mnestic functions; keloid inclinations; spondylosis 5 ankylopoetics; diseases of the movement organs due to wear and tear; sclerotic fundus; amyloidosis; morphea; fibrocytic mastopathy.

In veterinary medicine, the compounds of the invention have similar fields of application as in human medicine, ie. eg skin damage (peeling), wound healing and bone fractures.

The process according to the invention is characterized by the characterizing part of the claim.

Thus, one substitutes for β-substituted glutamine or as-paragin or substituted derivatives thereof. In this case, for example, one of the acid amide hydrogen atoms having an acid character is substituted by the method of the invention, for example, with metallic sodium and the compound thus obtained is converted by reaction with, for example, 2-bromoethanesulfonic acid sodium salt to a compound of the general formula I .

The method according to the invention is further elucidated in the following by way of example.

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Example 3.02 g (10 mmol) of carbobenzyloxy-L-glutamine sodium salt (Liebigs Annalen 640, 145, 1961) is dissolved in 50 ml of dimethylformamide.

To the solution is added an oil suspension containing 12 mmol sodium hydroxide and then heated for two hours to exclude the humidity. Then, 2.11 g (10 mmol) of bromethanesulfonic acid sodium in 50 ml of dimethylformamide are added dropwise to the solution and the mixture is heated for a further two hours. Then it is evaporated under vacuum and the residue is extracted with ether and dried. The dry compound is dissolved in water, placed on a column of MDowex "50 and eluted with water. The eluate is evaporated under vacuum and the residue is dried over potassium hydroxide. Crude carbobenzyloxy-γ-L-glutamyltaurine is purified, which is purified by paper electrophoresis carried out at pH 6. 5. The relative mobility of cysteic acid * is 1.05. · R + (n-Butanol / pyridine / glacial acetic acid / water 15: 10: 3: 12) = 0.57.

t r X

The biological effect of the compounds of the invention according to the invention is hereinafter illustrated in two biological test reports, A and B. In these test reports, the biological effect of different types of compounds of general formula I. The tested compounds 20 have all unprotected N -terminal and C-terminal groups, i.e.

R 2 is hydrogen and is hydroxy. Compounds of general formula I where different from hydrogen and / or where A 1 is different from hydroxy have the same biological effect as the corresponding unprotected amino acid derivatives, one having to expect the protecting groups to be cleaved in the organism.

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Biological Test Report A.

Study of γ-L-glutamyltaurine as in this report is called litoralon.

5 In the following Tables 1-6, the test results are shown as mean values + standard deviation with the number of brackets. Determination of whether there is a significant difference between control trials and trials in treated animals was done with Student's t-test.

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Table 1

Effect of Litoralon on vitamin A concentration in serum.

Group of Litoralone Vitamin A Concentration in Serum 15 µg / day_ I. Control - 28.3 + 0.7 (20) II. 0.1 41.9 + 1.0X (20) III. 0.3 32.9 + 1.1 ”(20) IV. 1.0 27.4 + 0.6 (20) 20: x P <0.001 xx P <0.01 20 Sprague Dawley rats (10 males and 10 females) weighing 180-200-200 g were orally treated with those listed in Table 1 daily do see litoralon in the form of an aqueous solution for a period of 8 days. On the 9th test day, the animals were sacrificed by decapitation and the blood was collected. The vitamin A concentration in the serum was determined by Neeld and Pearson's method.

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Table 2

The effect of litoralone and vitamin A on granule formation induced by implanted cotton balls

Group Dose Dry weight of 5 Vitamin Ax litoralon granuloma local local oral mg yg yg / day I. Control - - 52 + 1.0 (24) II. Control +

Solvent - - - 54 + 3.1 (8) III. 2 - - 64 + 2.5 (8) IV. 2 0.1 65 + 2.7 (8) V. - 0.1 73 + 2.9 (8) VI. 2 - 0.1 90 + 4.1 (8) 15 x Hoffmann La Roche

The difference is significant: between groups II and III at P <0.05, between II and V at P <0.001, and between V and VI at 20 P <0.01. The granule conversion was determined according to Lee et al

Male Sprague-Dawley rats weighing 110-120 g. The tampons were removed from the dorsolateral subcutaneous implants after 10 days and weighed after drying to constant weight at 65 ° C.

25 8

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DK 159267 B

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DK 159267B

Table 5

The effect of Litoralon on the metamorphosis of Rana dalmatina

Group Body length Tail length _mm_mm_

Control 45.9 + 0.2 31.4 + 0.2 (60) 5 Treated Animals 40.2 + 0.2X 26.7 + 0.2X (60) x Significance: P <0.001

The experiment was conducted with larvae of Rana dalmatia with a 30 day age, a body length of 20-25 mm, and with already 10 visible protrusive hind legs. Over a period of 30 days, the test animals were placed in tap water containing 0.5 µg / ml litoralon for two hours each day. The control group was placed in tap water without the addition of litoralone. The tails were measured on the 30th day.

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Table 6

Litoralone's effect on blood sugar levels

Group I. Study II. Study _mg% ._ mg% _ 20 Control 94 + 3.6 (10) 94 + 4.09 (10)

Treated animals 82.4 +3.8 (10). 81 + 1.32 (10)

Significance at I. study: P <0.05 Significance at II. study: P <0.01

Male CGY white body rats 160-180 g were used for the experiment. The animals were kept on a standard diet. Blood samples were taken on the 5th day of testing after 18 hours of fasting. The blood glucose assays were performed according to the method of E. Hultman. Litoralon was given orally at daily doses of 1 µg / kg.

30 11

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Biological Test Report B.

Study of synthetic γ-aspartyl taurine and its derivatives.

1. β-Aspartyl-N-methyltaurine 5 a) Effect on blood sugar level Control 107 mg%

Treated animals 93 mg%

Significance: P <0.05

20-20 rats were used for the samples. Measurements were carried out -10 after 18 hours of fasting. The dose used was 1 µg / kg body weight for 4 days in the form of a solution by oral administration.

b) Effect to increase serum vitamin A level

Oral dose Evoked vitamin A yg% 15 yg / 200 g body weight 0 (control) 8.5 5 11.0 1 11.5 0.3 12.5 20 0.1 16.1 0.05 14.8 0.01 12.5 0.005 10.5

Significance: P <0.01 25 For the experiments, 20-20 Wistar male rats weighing 200-220 g were used.

Trial period: 6 days.

c) Effect on blood silicon level: on

Silicon (mg / g blood) __0 hours_20 days_40 days

Control group 0.110 + 0.004 0.120 + 0.010 0.154 + 0.015

Treatment Group I 5 µg / day 0.100 + 0.005 0.315 + 0.014XX 0.345 + 0.015 Treatment Group II 10 µg / day 0.107 + 0.009 0.370 + 0.119 0.360 + 0.017 xx Significance: P <0.001 12

DK 159267 B

The results are significant from the 13th day at the significance level P <0.01 and from the 20th day at the level P <0.001. The experiments were performed on inbred male rabbits of body weight 2.5-3 kg. The active ingredient was administered in the 5 daily doses shown in the table. The determination of silicon was performed according to Gaubatz's method (Gaubatz E., Clin. Wschrft. 14, 1753, 1935) in 5 ml of blood samples taken from the animals' ears.

d) The effect of β-aspartyl-N-methyl-taurine and vitamin A on the 10 granulogenic effect caused by implantation of cotton wool;

Group Dose Weight of dry

Vitamin Ax g-aspartyl-N- rf 9ranuloma methyltaurine local local oral mg ug u g / day ---: -

Control I. - - - 54 + 1.7

Control II. + solvent - - - 55 + 3.4

Treatment Group III. 2 - - 66 + 12.5

Treatment Group IV. 2 0.1 - 67 + 2.6 Treatment Group V. - - 0.1 79 + 2.8

Treatment Group VI. 2 - 0.1 96 + 4.4 x Hoffmann la Roche

The differences are significant as follows: 25 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.

The determination of granuloma was performed on male Sprague-Dawley body weights 110-120 g by the method of Lee et al (Lee K.H., Fu Ch.Ch., Spencer M.R. Tong T.G. and Poon R.J., Pharm.

30 Sci., 62, 895, 1973). The dorsolaterally subcutaneously implanted tampons were removed after 10 days and measured after drying at 65 ° C to constant weight. 1 P- Aspar tyl-homotaurin ·.

35 a) Effect on blood sugar level

Control group 105 mg%

Treated group 94 mg%

Significance, number of test animals and test method were as indicated in section. 1 a) above.

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b) Increasing effect on serum vitamin A level

Oral dose. Induced vitamin A

µg / 200g body weight µg% _ 0 (control) 8> 6 5 5 12.0 1 13.5 0.3 14.0 0.1 15.8 0.05 15.0 10 0.01 12.0 0.005 _10,0_

Significance, number of test animals and test method were as indicated above under section. Ib).

C) Effect on blood silicon level:

Silicon (mg / g blood) _0 hours _20 days_40 days_

Control group 0.104 + 0.009 0.134 + 0.015 0.157 + 0.020

treatment Group

20 I. 5pg / day 0.094 + 0.007 0.309 + 0.014XX 0.340 + 0.014XX

Treat 1 incrscnr up

II. 10 µg / day 0.109 + 0.010 0.372 + 0.120XX 0.363 + 0.018XX

Significance, number of test animals, arrangement and method of determination as indicated above under cl. 1 c).

D) The effect of β-aspartyl homotaurine and vitamin A on the granulomatous effect of cotton implantation;

Dose Weight of dried

Vitamin Ax β-aspartyl granuloma local homotaurin g mg local oral _ µ g g / day

Control group I - - 52 + 1.5

Control group II - - - 53 + 3.2 + solvent 35 Treatment group III. 2 - - 64 + 12.3

Treatment Group IV. 2 0.1 - 65 + 2.4

Treatment group V. - - 0.1 77 + 2.6

Treatment group VI._2_ 0.1_94 + 4.2_ 14

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x Hoffmann la Roche

Significance, number of test animals, arrangement and method of determination were as indicated above under point 1 (d).

Claims (1)

20 I .R CO-N + X Me 1 1 wherein A, R, R and n have the meanings given above and Me is an alkali metal, is alkylated with an alkali metal m-halo-C if desired is converted to a physiologically acceptable salt or released from a salt and / or if desired prepared in optically active form by splitting the racemic product obtained. For: Chinoin Gyogyszer és Vegyészeti Termékek Gyåra RT. fENERET