CH621334A5 - - Google Patents

Description

621334

2nd

PATENT CLAIM Process for the preparation of new amino acid derivatives of the general formula

RI-HN-CH-CO-A1

I.

(CH2) n (I)

I.

CO — N— (CH) - (CH ;,) t — B1

I I

R R2

in the form of their racemates or optical antipodes,

wherein

A1 for a group of the general formula - (NH — CH — CO) r — Y

I.

R3

in which Y is hydroxy or alkoxy having 1 to 4 carbon atoms,

r is an integer of at most 2000 and R3 is a group of the general formula

- (CH,) n — CO — N— (CH) - (CH2) t — B1

I I

R R2

mean,

B1 for -SOoOH; -0S020H or -OPO (OH) 2,

R for hydrogen, alkyl with 1 to 4 carbon atoms, R1 for hydrogen, alkoxycarbonyl, aralkoxycarbonyl,

aralkoxycarbonyl substituted by halogen, alkoxy or nitro,

R2 for hydrogen, carboxyl, alkyl with 1 to 4 carbon atoms, alkoxycarbonyl or carboxamide,

n is an integer from 1 to 3 and t is an integer from 1 to 2,

and of their salts, characterized in that a peptide derivative of the general formula

R1-NH-CH-CO-A3

I.

(CH2) n (II)

CO-A2

wherein R1 and n have the above meaning, A3 is a group of the general formula

- (NH-CH-CO) r-Y

I.

R4

means in which Y and r have the meaning given above, R4 represents a group of the general formula - (CH2) n-CO-A2 and A2 represents p-nitrophenoxy, pentachlorophenoxy or C2- to C4-alkoxycarbonyloxy, with a compound of the general formula

HN— (CH) - (CH2) t — B2 (III)

I I

R R2

wherein R, R2 and t have the meanings defined above and B2 -S020H; -0S020H; -OPO (OH) 2 or the mercapto-

represents group, implemented, an optionally obtained compound in which B2 is the mercapto group, to an amino acid derivative of the general formula (I) in which A1, R, R1, R2, n and t have the abovementioned meaning and B1 for the group - SOaOH stands, is oxidized, optionally a compound of the general formula (I) obtained is converted into a salt, or is released from a salt.

The present invention relates to the process for the preparation of new amino acid derivatives of the formula I, in which the symbols have the meaning given there.

Some of these compounds have valuable pharmacological properties. Still other of the compounds accessible according to the invention are intermediaries for the production of substances with valuable biological or pharmacological effects. All of the compounds according to the invention are new.

Among the compounds mentioned in CH-PS 617 183, y-L-glutaryl-taurine, which has the formula (XXIV)

h2h-ch-cooh I

ch2

i (xxiv)

ch2 I

CO-NH-CH2-CH2-S020H

corresponds to and has a broad therapeutic and preventive spectrum of effects for indirect or directly attributable to violations of the "AGAS" (aerobiospheric genetic adaptation system).

To explain the term “AGAS”, the most important tissues and organs that make up this system are listed below.

a) All biological interfaces that are in contact with the outside air as a biosphere (skin and skin structures, cornea and conjunctiva, oral and pharynx cavities, airways and lungs);

b) skeleton and limbs (long bones and spongy bones, ball joints, synovial membrane, skeletal muscles);

c) the organs involved in the regulation of the ion balance (transepithelic transport system: villi and kidney channels);

d) The theko-donte (fixed in the tooth socket by the root) the bite required for the comminution of the food;

e) Hearing, smell and voice organs.

The compounds accessible according to the invention thus have a beneficial biological or therapeutic effect on the organs or tissues of the AGAS system listed here.

The compounds accessible according to the invention also act on the following functions associated with the AGAS system: radiation protection effect, favoring wound healing, generally activating the mesenchyme, protection against the constantly increasing risk of infection and contamination of the skin and mucous membranes (lysozyme generation of the moist Mucous membranes, activation of ciliated epithelia in the respiratory tract, etc.), gei

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increased protection against infections caused by viruses and fungi.

The compounds according to the invention are effective against the constantly and greatly increasing stress effects (e.g. meteorological influences, strong differences between day and night temperature, increased risk of injury) by stabilizing the adaptation syndrome and at the same time warding off the peripheral tissue damage of the glucochorti-coids (thus Example damage to the connective tissue, injuries to the bone matrix stock, etc.). Development of immune homeostasis (increased knowledge of the body which cells are in the body and which are not).

The compounds accessible according to the invention exert their effect partly directly, partly via the control of the vitamin A metabolism, by the production of vitamin A metabolites of a more polar character. This effect is comparable to that of the parathyroid hormone on the 25-hydroxy-chole-calciferol-la-hydroxylase enzyme of the kidney channels. The broad pharmacological, biochemical and therapeutic effects of the compounds according to the invention can be understood from this explanation. The directions of action of the connections are as follows:

A) Effects with vitamin A character:

a) Pharmacological and biochemical effects: Labeled sulfates are incorporated to an increased extent in the rat's cartilage or the eye lens, the liver and lung tissue of the chicken embryo; radioactively labeled phosphorus is incorporated into the rat's sword cartilage to an increased extent; the chon-droitin sulfate synthesis increasing effect; beneficial effect on wound healing or experimentally worsened wound healing in rats and dogs by administration of chortisone; the potentiating effect of vitamin A in hypo and hypervitaminoses experimentally caused in rats and chickens; the effects moderating the ulcer-related stress effects in rats; beneficial effect on degranulation of mastocytes; increasing effect on the production of Lyso-sym; Effects on the household of trace elements (silicon, zinc, copper, manganese, fluorine); promoting effect on epithelial formation; increasing effect on alkaline phosphatase activity; the effect exerted on the granuloma sack formation caused by local action of vitamin A; the very flat course of the dose-response curve or the change in the sign of the effect at large doses. Activating effect on the Golgi apparatus; beneficial effect on the formation of calyx cells; increasing effect on the concentration of vitamin A;

b) clinical-therapeutic effects: keratocunjunctivis sicca; Sjogren's syndrome; rhino-laryngo-pharyngitis sicca; Ozaena; Chronic bronchitis; Sinobronchitis; Mucovisci can; constitutional lung diseases in young children; Paradentosis; Infection of the skin and mucous membranes for viruses and fungi; chortisone-antagonistic effect; beneficial effect on the Heii process in surgical wounds and mucosal wounds; erosio colli; pruritus-like; Reduced sense of smell and taste.

B) Effects without a vitamin A character:

a) Pharmacological and biochemical effects: effect on the blood sugar level in the sense of a temporary decrease; increasing effect on the phosphaturia, lowering effect on the phosphorus level in the

621334

Serum; radioprotective effect; in the case of labyrinth experiments with inactive animals, the time-reducing effect required to achieve the goal; reducing effect on experimentally induced fluorine and cadmium toxicosis; cyclic adenosine monophosphate emptying of the kidney; the symptoms of experimentally induced lathyrism moderate effect; Reduction in sensitivity to histamine; increasing effect on the activity of the liver enzyme tyro-sinaminotransferase.

b) Therapeutic effects: weak radiation damage; Vitiligo; Muscle hypotension; psychoenergetic effect; beneficial effect on involutional and gerontological conditions as well as on mnestic functions; cheloidal tendencies; Ankylopoetic spondylosis; wear and tear diseases of the locomotive organs; sclerotic fundus; Amyloidosis; Morphes; fibrocystic mastopathy.

When the compounds obtainable according to the invention are administered, the duration of the treatment is extremely different. Some diseases (e.g. rhinoceronary pharyngitis sicca) become symptom-free after just two weeks after oral administration of 3X5 micrograms daily; symptomatic improvement of other diseases (e.g. paradentosis, Sjögren's syndrome) takes one to two months, while others require Diseases (e.g. spondylosis ankylopoetica) need to be treated for a quarter to half a year.

Any pharmaceutical, preventive, cosmetic or veterinary preparation can be produced in a simple manner from the compounds obtainable according to the invention. The preparations can contain a single active ingredient or a combination of active ingredients. The dose of pure active ingredient is 50-500 nano-grams per day and kilo of body weight and is administered in three separate doses.

One tablet contains 2-20 micrograms, preferably 10 micrograms of active ingredient and also biologically inert carriers (e.g. milk sugar, starch) as well as the usual tableting aids (granulating and lubricating agents such as polyvinylpyrrolidone, gelatin, talc, magnesium stearate, Aerosil etc.) ).

Because of the extraordinarily small dose, it is advisable to add the active ingredient in the form of a solution to the tablet mass before granulation. In this way, the active ingredient is distributed evenly. Incidentally, the low active substance content enables the active substance to be produced on a large laboratory scale even if many millions of tablets are produced, and at a price that is acceptable to every patient. The active ingredients are stable, so the tablets can be put on the market without specifying a time limit for consumption. The active substance content of delayed acting tablets or capsular capsules can be 10-20 micrograms. In the case of injectable preparations, the appropriate dose per ampoule is 5-10 micrograms, and the preparation can, if desired, be prepared as a powder ampoule in which the active ingredient is mixed with an indifferent, water-soluble excipient. Parenteral administration can be carried out intramuscularly, subcutaneously or intravenously. In the specified concentration, the active ingredients do not damage tissue or vessel walls. The active ingredients can also be administered as an infusion. Sup-positorien contain 2-20, preferably about 10 micrograms of active ingredient and are made from cocoa butter or from synthetic waxes or fats suitable for this purpose (for example those produced in the Federal Republic of Germany

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Imhausen mass). The active substance content of cosmetic ointments used to heal the skin is 0.1-1 micrograms / g. The ointment base can be hydrophilic or hydrophobic and contains the usual components: for example cholesterol, paraffin, glycerin, lanolin, cocoa butter, linseed oil etc. The active ingredients can also be formulated as aerosol preparations, the active ingredient content also being 0.1-1 micrograms / g . When formulated as a perlingual tablet, a tablet contains 10 micrograms of active ingredient, the tablet's disintegration time is Yi-1 hour, polymers for the sustained release of the active ingredient can be prepared, for example, as a suspension and contain 1-5 micrograms of active ingredient / g polymer. Injection preparations with retarder effect can be formulated either using high molecular weight polymers or from the salts of the compounds according to the invention formed with high molecular weight organic bases (for example histone, protamine), one ampoule containing 10-20 micrograms of active ingredient. Powders for cosmetic or skin healing purposes are prepared with the usual carriers (for example talc) and contain 0.1-1 g active ingredient / g powder. For ophthalmology, the compounds are formulated as drops or as ointments that are miscible or immiscible with the tear fluid; the active substance content of these preparations is 0.1-1 microgram / g. Children should be administered the compounds according to the invention in a dosage of about 0.3 micrograms per kilogram of body weight.

The sterile preparations are expediently produced by sterile filtration.

The desired preventive pharmacological or cosmetic effect of the preparations listed above can be increased and supplemented by numerous combinations. The following are explicitly mentioned as possible biologically active additives and their use is to be protected:

Vitamins A, C, E and K, trace elements, chortisone and its derivatives, progesterone, thyroid hormones, radio-mymetically and immunosuppressive substances, psy-chopharmaca, especially tranquillizers, timoleptics, organic silicon compounds, gerontological preparations, which lower the blood cholesterol level Substances, oral antidiabetics, anti-inflammatory substances, antihistamines, etc. The dosage of these active additives is generally the same as the usual therapeutic dose when the substances are used alone.

The compounds accessible according to the invention can also be used as an additive to nutritional or pharmaceutical premixes. They partly cause weight gain, partly they reduce the need for vitamin A or improve its metabolism. Furthermore, the absorption of the trace elements is increased by the compounds, their level in the blood is raised. When the compounds are used as an additive to animal feed, the dosage orally is expediently 200 nanograms / kg and day. When mixed into the feed, this corresponds approximately to a concentration of 1-2 micrograms / kg or 1-2 mg / ton feed, i.e. a concentration of 0.001-0.002 ppm. In view of these very low concentrations, use as a feed additive is extremely economical. The compounds are preferably admixed with vitamin premixes or used in microcapsules which contain the compounds according to the invention in addition to other necessary feed additives. The compounds can also be used when soaking with drinking water, in leaking salt or occasionally as a spray.

In veterinary medicine, the compounds according to the invention have fields of application similar to those in human medicine, i.e. for example skin damage (peeling), wound healing, broken bones etc.

It is a common structural property of all compounds of the general formula (I) that they contain a dicarboxylic acid substituted in the a-position or its derivative which is also substituted in other places via its w-carboxyl group with an acid amide bond bonded to a primary or secondary alkylamine, which contains a group of strongly acidic character apart from various substituents on its alkyl side chain in the «position.

example 1

0.52 g (4 mVal) of a-poly-L-glutamic acid (degree of polymerization 80) (Acta Chim. Acad. Sei. Hung. 5, 267, 1955) are dissolved in 10 ml of dimethylformamide, the solution is cooled with ice water. 1.39 g (10 mmol) of p-nitrophenol and 0.82 g (4 mmol) of dicyclohexylcarbodiimide are added with stirring. After 10 minutes, dicyclohexylcarbamide begins to separate out. The mixture is stirred at room temperature for one day, then the dicyclohexylcarbamide is filtered off, the unreacted dicyclohexylcarbodiimide is also converted to dicyclohexylcarbamide by addition of 0.3 ml of glacial acetic acid and this is also filtered off. The solution is poured into a mixture of 100 ml of ether, 100 ml of petroleum ether, 20 ml of ethyl acetate and 2 ml of glacial acetic acid. The precipitated product is centrifuged off, washed several times with ether and then dried in vacuo over sulfuric acid. 0.80 g of a-poly-L-glutamic acid active ester 1 is obtained, the p-nitrophenolate content of which is 2.6 meq / g.

For the characteristic groups, the infrared spectrum has the following maxima: NH 3300 cm-1; C = 0 (COONP) 1765 cm-1.

0.25 g of a-poly-L-glutamic acid-y-p-nitrophenyl ester 1 are dissolved in 7 ml of pyridine. While stirring, 0.125 g (1 mmol) of taurine in 1 ml of water and 0.28 ml (2 mmol) are added in ten portions so that the reaction always takes place in clear solution. Another ml of water is required. The reaction mixture is left to stand at room temperature for three days, then evaporated in vacuo and the residue dried. After thorough digestion with ether, the residue is ground to a powder. After dissolving in water and then lyophilizing, 0.20 g of a-poly-y-glutamyl taurine 1 is obtained. The chromatogram shows contamination by 0.4% taurine.

Analysis: S 10.1%.

For the characteristic groups, the infrared spectrum has the following maxima: OH 3100-3400 cmr1 (broad); C = 0 (amide I) 1650 cm-1; C = 0 (Amide II) 1550 cm "1; S = 0 (S020H) 1220.1040 and 600 cm" 1.

Example 2

0.26 g (2 mVal) of a-poly-L-glutamic acid (degree of polymerization 580) (J.A. C. S. 80, 4631, 1958) are swollen and dissolved in 15 ml of dimethylformamide. 0.69 g (5 mmol) of p-nitrophenol and 0.41 g (2 mmol) of dicyclohexylcarbodiimide are added to the solution with stirring. The reaction mixture is stirred at room temperature for two days. 0.30 g of a-poly-L-glutamic acid active ester 2 is obtained.

For the characteristic groups, the infrared spectrum has the following maxima: C = 0 (COOH) 1720 cm-1. The remaining bands are similar to those described in Example 20, but wider. From the a-poly-L-glutamic acid-y-p-nitrophenyl ester 2, 0.27 g of lyophilized a-poly-y-L-glutamyl taurine 2 is obtained in the manner described in Example 20. The contamination of the product by taurine is less than 0.4% according to chromatographic studies.

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621334

Analysis: S 7.3%.

The infrared spectrum is identical to that described above.

Example 3

0.26 g (2 mVal) of a-L-polyglutamic acid (degree of polymerization 580) are dissolved in 8 ml of dimethylformamide. The solution is cooled to -10 ° C in a cold mixture of ice and table salt. After the addition of 0.28 ml (2 mmol) of triethylamine, the solution gels and does not become liquid again even with the addition of a further 8 ml of dimethylformamide and intensive stirring. 0.28 ml (2 mmol) of isobutyl chlorocarbonate and, after 30 minutes of activation, 0.16 g (2 mmol) of cysteamine in 2 ml of dimethylformamide are then added dropwise. The reaction mixture is stirred at -5 ° C for two hours, at room temperature for four hours and then poured into a mixture of 50 ml of chloroform and 50 ml of petroleum ether. The precipitated white precipitate is centrifuged off, washed several times with a mixture of chloroform and petroleum ether, swollen a few times in alcohol and then precipitated with ether. 0.33 g of a-poly-y-L-glutamyl-s cysteamine is obtained.

Analysis: S 14.7%.

0.16 g of a-poly-y-L-glutamylcysteamine are suspended in 5 ml of glacial acetic acid and then mixed with 1 ml of 30% hydrogen peroxide. The reaction mixture is left at room temperature for three days, the solution slowly clearing. It is diluted with water and then lyophilized. This gives 0.20 g of white a-poly-y-L-glutamyl taurine, the chromatographically determined tau rin impurity of which is 0.5%.

15 Analysis: S 11.9%.

The infrared spectrum agrees with that already described.

v