CS209861B2 - Method of making the new derivatives of the amino acids - Google Patents

Description

The invention relates to a process for the preparation of the novel amino acid derivatives of the general formula (I) below, some of which have valuable pharmacological properties, others are intermediates for the production of substances with advantageous biological or pharmacological effects. All compounds obtained by the process of the invention are novel.

The compounds which can be prepared by the process according to the invention can be represented by the general formula I,

H 2 N - CH - CO - A ¹ (CH ₂ ) i

CO-N (CH ₂₎ t Bl

R (I) wherein is

A ¹ - (NH-CH-CO) _r -Y,

R ³ in which

Y represents a hydroxyl radical or a (C 1 -C 4) alkoxy radical, r denotes an average degree of polymerization of at most 2000,

R ³ represents a group of formula - (CH2) n-CO-N _{(CH2 t} j -B ⁴

R in which

B ⁴ represents a group of the formula —SO2 OH, —OSOOH or —OPO (OH) ₂ ,

R is hydrogen or C1-C4alkyl, n is an integer from 1 to 4 and t is an integer from 2 to 6.

The invention also includes a process for the preparation of pharmaceutically acceptable salts of said compounds or optically active antipodes thereof.

The compounds prepared by the method of the invention are characterized by a broad spectrum of therapeutic and preventive activity against pathological changes, having a direct or indirect cause in damage to the aerobiospheric genetic adaptation system (AGASJ).

The most important tissues and organs forming this system are listed below to clarify the concept of AGAS:

aj All biological boundary surfaces in contact with outside air, such as the biosphere (skin and skin, cornea and conjunctiva, oral and esophagus, airways and lungs), bj skeleton and limbs (hollow tubular bones and spongy bones, spherical joints, lining of the joint cavity, skeleton muscles],

c) bodies involved in the regulation of ion management (trans-epithelial transport system of the intestinal villi, renal ducts),

d) dentition, necessary for grinding food and fixed roots in dental beds,

(e) auditory, olfactory and vocal organs.

The compounds prepared according to the invention thus have a favorable theological or therapeutic effect on the organs or tissues of the AGAS system mentioned above.

The compounds prepared by the method according to the invention furthermore have the following AGAS-related functions: radiation protection, wound healing effect and generally activating mesenchym, protection against the ever increasing risk of infection and contamination of the skin and mucous membranes (formation of wet mucosal lysozyme, activation ciliated lining in the airways, etc.), increased protection against virus and fungal infections.

The compounds prepared by the process of the invention are effective against increasingly and substantially with increasing stress effects (e.g., meteorological effects, considerable differences between day and night temperature, increased risk of injury) on shore life by stabilizing adaptation syndrome while averting damage to peripheral tissues by glucocorticoids (for example, damage to the connective tissues, damage to the underlying bone mass, etc.) Emergence of immunohomoeostasis (an increased ability to recognize the body which cells belong to the body and which do not).

Said compounds act in part directly, in part by controlling the metabolism of vitamin A, by producing vitamin A metabolites of a more polar nature. This effect is comparable to that of parathyroid hormone on the 25-hydroxycholecalblferol-1-a-hydroxylase enzyme of the renal tubules. By this explanation the broad pharmacological, biochemical and therapeutic effect of the compounds prepared by the process according to the invention becomes understandable. The areas of action of these compounds are as follows:

A. Vitamin A effects

(a) Pharmacological and biochemical effects Sulfates labeled with the radioactive element are increasingly incorporated into the bone cartilage of the rat, optionally into the lens, liver and lung tissue of the chicken embryo; radiolabeled phosphorus is increasingly incorporated into the rat's bone scalp; chondroitin sulfate synthesis enhancing effect; a beneficial effect on wound healing, experimentally impaired administration of cortisone in rats and dogs; vitamin A potentiating effect of hypovitaminoses or hypervitaminoses, experimentally induced in rats and chickens; sedative effect on stress effects caused by rat ulcer; a beneficial effect on mast cell degranulation; lysozyme production-enhancing effect; effects on the management of trace elements (silicon, zinc, copper, manganese, fluorine); a beneficial effect on epithelial formation; escalating effect on alkaline phosphotase activity; action on granulon vesicle formation induced by the local effect of vitamin A; very flat course of the dose / effect curve or change of the effect sign at large doses; activating effect on Golgi; a favorable effect on the formation of calyx cells; Vitamin Concentration Increasing Effect

AND.

b) Clinical therapeutic effects

Keratoconjunctivitis sicca; SjOrgren's syndrome; rhinolaryngopharyngitis sicca; ozaen.a; chronic bronchitis; sinobronchitis; mucoviscidosis; constitutional lung disease in young children; periodontitis, susceptibility of the skin and mucosa to infection by viruses and fungi; cortisone antagonist; beneficial effect on wound healing of skin and mucosa during surgery; erosio colli; itch-induced decrease in olfactory and taste sense.

B. Vitamin A-free effects as well as Pharmacological and Biochenk effects

Effect on blood sugar in terms of transient reduction; escalating effect on phosphaturia; reducing the effect on serum phosphorus; radio-protective effect; an effect reducing the time needed to reach the target in experiments with the labyrinth of inactive animals; a dampening effect on experimentally induced fluorine and cadmium toxicosis; adenosine monophosphate-induced cyclic renal emptying effect; a mitigating effect on the symptoms of experimentally induced lathyrism; decreased sensitivity to histamine; enhancing effect on liver enzyme tyrosine aminotransferase activity.

bj Therapeutic effects

Slight radiation damage; local lack of dye in the skin; muscular hypotonia; psychoenergetic effect; a beneficial effect on involutionary and gerontological states as well as on mnestic functions; tendency to scar fibroma formation; spondylosis ankylopoetica; musculoskeletal disorders caused by wear; sclerotic fundus; amyloidosis; morphea; fibrocystic mastopathy.

WITH

When administering the compounds of the invention, the treatment time varies greatly. In many diseases (such as rhinolaryngopharingytis sicca), oral administration of 5 µg three times a day does not show symptoms after only two weeks, to symptomatically heal others. Diseases (eg periodontitis, Sjorgren's syndrome) are required for one to two months, and for other diseases (eg spondylosis ankylopostica), treatment must take place for a quarter to a half year.

Any of the pharmaceutical preventive, cosmetic or veterinary medicinal preparations can be prepared in a simple manner from the compounds prepared by the process according to the invention. These preparations may contain a single active ingredient or a combination of active ingredients. The dose of pure active ingredient is 50-500 [mu] g per day, per kg body weight, and is administered in three divided doses.

One tablet contains 2 to 20 µg, preferably 10 µg of active ingredient, and additionally theologically inert carriers, for example milk sugar, starch, as well as conventional tabletting aids (granulating and glidants, for example polyvinylpyrrolidone, gelatin, talc, stearate magnesium, aeroeil, etc.).

Because of the extremely low dosages, it is expedient to add the active ingredient to the mass from which the tablets are then compressed, prior to granulation, in the form of a solution. In this way, an even distribution of the active substance is achieved. Moreover, the low active substance content makes it possible to produce the active substance - even in the manufacture of many millions of tablets per year - on a laboratory scale only. at a price that is available to every patient. Since the active substances are stable, the tablets can be supplied to the market without a expiry date. The active ingredient content of retarding tablets or spansular capsules may be in the range of 10 to 20 µg. For injectable preparations, the effective dose per vial is 5 to 10 µg, and the preparation can optionally be prepared in the form of a powder ampoule in which the active ingredient is mixed with an inert water-soluble filler. Parenteral administration can be performed intramuscularly, subcutaneously or intravenously. At the indicated concentration, the active compounds do not have a detrimental effect on either the tissue or the vessel walls. The active ingredients may also be administered as an infusion. Suppositories contain from 2 to 20, preferably about 10, g of active ingredient and are made from cocoa butter or synthetic waxes or fats suitable for this purpose. Content of active substance in cosmetic or ointment. skin healing amounts to 0.1 to 1 pg / g. The ointment matrix may be hydrophilic or hydrophobic and contains conventional ingredients, for example, cholesterol, paraffin, glycerin, lanolin, cocoa butter, linseed oil, etc. The active ingredients may also be formulated as aerosol preparations, the active ingredient content being also in the range of 0, 1 to 1 pg / g. In the form of a perlingual tablet, the tablet contains 10 µl of active ingredient, the disintegration time being 0.5 to 1 hour. Sustained release polymers can be prepared, for example, as a suspension and contain 1 to 5 µg of active ingredient per g of polymer. Delayed injectable preparations may be formulated using either high molecular weight polymers or salts of the compounds of the invention formed with high molecular weight organic bases (e.g., histone, protamine), each ampoule containing 10 to 20 μξ of the active ingredient. Powders for cosmetic or skin treatment are prepared with conventional carriers, for example talc, and contain 0.1 to 1 µg of active ingredient per g of powder. For ophthalmology, the compounds of the invention are formulated as drops, optionally as ointments, miscible or immiscible with tears; the active ingredient content of these preparations is 0.1 to 1 µg / g. In children, the compounds of the invention should be administered at a dose of about 0.3 µg per kg body weight.

Sterile preparations are conveniently prepared by sterile filtration.

The desired preventive pharmacological or cosmetic effect of the above-described preparations can be increased and supplemented by numerous combinations. Among the biologically active additives that may be considered are, in the first place, the following substances and the use thereof:

Vitamins A, C, E and K, trace elements, cortisone and its derivatives, progresterone, thyroid hormones, radiomimetics and immunosuppressive agents, psychopharmaceuticals, especially soothing agents, timoleptics, organic silicon compounds, gerontological agents, lowering agents blood cholesterol levels, oral antidiabetics, anti-inflammatory agents, antilrstamines, etc. The dosage of these active ingredients is generally the same as the usual therapeutic dose using these agents alone.

The compounds produced by the process according to the invention can also be used as an additive in nutritional or medicinal mixtures. They cause weight gain, reduce the need for vitamin A or improve its metabolism. Further, the compounds of the invention increase the resorption of trace elements and their blood levels. When the compounds of the invention are used as feed additives, the oral dose is expediently 200 µg / kg per day. When mixed with the feed, this dose corresponds to a concentration of approximately 1 to 2 g / kg or 1 to 2 mg / t of feed, i.e. a concentration of 0.001 to 0.002 ppm. Because of these very low concentrations, their use is as. feed additives extremely economical. Preferably, these compounds are added to the vitamin masterbatch. or are used in microcapsules containing the compounds of the invention together with the necessary feed additives. The compounds according to the invention can furthermore be used in drinking water for cattle feeding, in licking salts or, if desired, in the form of a spray.

In veterinary medicine, the compounds of the invention have similar fields of use as in human medicine, e.g., in skin diseases, wound healing, bone fractures, etc.

A common feature of the structure of all compounds of formula I is that they contain a dicarboxylic acid substituted at the α position, or a derivative thereof substituted at other positions, linked via its ω-carboxyl group by bonding the acid amide to a primary or secondary alkylamine which The substituent contains a strongly acidic group on its side alkyl chain in the ω-position.

The process according to the invention for the preparation of the compounds of the formula I, their salts and optical antipodes is characterized in that the compounds of the formula II are:

HzN - CH - CO - A ³ (CH 2)

CO — A ² (II) where n is an integer from 1 to 4,

A ³ is - (NH-CH-CO), -Y,

R ⁴ in which

Y represents a hydroxyl radical or a (C1 -C4) alkoxy radical, r denotes an average polymerization degree of at most 2000,

R ⁴ represents a group of the formula - (CH 2 -CO-A ² , where n is as defined above and Az represents a p-nitrophenoxy, pentachlorophenoxy or C 2 -C 4 alkoxycarbonyloxy group, reacted with a compound of formula III ,

HN - (CH ^{2 -} - B ²

R (III) wherein

R and t are as defined above and B ² is -SO 2 OH, OSO 2 OH or OPO (OH) 2 or mercapto, and when B ² is mercapto, the resulting compound is oxidized and optionally converted released into the salt and / or prepared in optically active form using optically active starting materials or by resolution of the racemic mixture formed as the product.

The process according to the invention is prepared from peptides containing α-amlnocarhoxylic acids, for example α-glutamylglycine, derivatives activated on the ω-carboxyl group, for example p-nitrophenyl esters, and the compounds thus obtained acylate taurine, homotaurine or cholamine phosphate.

Polymeric or oligomeric derivatives can also be prepared from the compounds of formula (I). They are obtained by reacting ω-active esters of n-polyaminodicarboxylic acids - for example χ-p-nitrophenyl ester of α-poly-L-glutamic acid - with taurine, homotaurine or cystamine (in the case of cystamine, oxidation is also necessary). Conversely, if desired, these polymers can be degraded to the monomeric compounds of formula I as described above.

To prepare salts of the compounds of the invention, the compound of formula I is reacted with an alkali or alkaline earth metal hydroxide or carbonate or an organic base.

The process according to the invention is illustrated by the following examples.

He did

0.52 g (4 mol of α-poly-L-glutamic acid (polymerization step 80) (Acta Chim. Acad. Sel. Hung. 5, pp. 2-7, 1955) is dissolved in 10 ml of dimethylformamide and cooled. With stirring, 1.39 g (10 mmol of p-nitrophenol) and 0.82 g (4 mmol) of dicyclohexylcarbodiimide were added, and after 10 minutes dicyclohexylcarbamide began to precipitate, and the mixture was stirred at room temperature for one day, then precipitated. The dicyclohexylcarbamide was filtered off and the unreacted dicyclohexylcarbodiimide was reacted to dicyclohexylcarbamide with the addition of 0.3 ml glacial acetic acid, which was also filtered, and the solution was poured into a mixture of 100 ml ether, 100 ml petroleum ether, 20 ml ethyl acetate and 2 ml glacial acetic acid. The product is centrifuged, washed several times with ether, and then dried with sulfuric acid under reduced pressure to give 0.80 g of active ester 1 (α-poly-L-glutamic acid containing 2.6 mval / g of p-nitrotopheno). látu.

The infrared spectrum is characterized by the following maxima for the characteristic groups:

NH 3300 cm ^-1 ;

C = O (COONP) 1765 cm -1;

C = O (amide IJ 1660 cm ^-1 ;

C = O (amide IIJ 1500 cm ^-1 ;

NO 2 1530 and 1360 cm ⁴ .

0.25 g α-poly-L-glutamic acid β-p-nitrophenyl ester 1 is dissolved in 7 ml pyridine. 0.125 g (1 mmol) of taurine in 1 ml of water and 0.28 ml (2 mmol) in 10 portions are added to the solution while stirring, so that the reaction always proceeds in a clear solution. To this must be added another ml of water. The reaction mixture was allowed to stand for three days at room temperature, then evaporated under reduced pressure and dried. After thorough digestion with ether, the residue is triturated to a powder. After dissolution in water and subsequent lyophilization, 0.20 g of α-poly-γ-glutamyltaurine 1 is obtained. The chromatogram shows taurine contamination at 0.4%.

Analysis: 10.1%.

The infrared spectrum is characterized by the following maximums for characteristic groups:

OH 3100—3400 cm ^-1 (wide);

C = O (amide I) 16.50 cm ^-1 ;

C = O (amide II) 1550 cm ^-1 ;

S = O (SO 2 OH) 1220, 1040 and 600 cm ^-1 .

Example 2

0.26 g (2 mval) of α-poly-L-glutamic acid (polymerization step 580) (J.A.C. 80, p. 4631, 1958) was swollen and dissolved in 15 ml of dimethylformamide. 0.6.9 g (5 mmol) of p-nitrophenol and 0.41 g (2 mmol) of dicyclohexylcarbodiimide were added to the solution with stirring. The reaction mixture was then stirred at room temperature for 2 days. 0.30 g of active .alpha.-poly-L-glutaronic acid ester 2 is obtained.

The infrared spectrum is characterized by the following maximums for characteristic groups:

C = 0 (COOH) 1720 cm ^-1 ;

The other belts are similar to those described in Example 1, but are wider.

From α-poly-β-nitrophenyl ester 2 of α-poly-L-glutamic acid, 0.27 g of lyophilized α-poly-χ-L-glutamyltaurine 2 is obtained as described in Example 1. %.

Analysis: 7.3%.

The infrared spectrum of this product is identical to that described above.

Example 3

0.26 g (2 mol) of α-poly-L-glutamic acid (polymerization step 580) was dissolved in 8 ml of dimethylformamide. The solution was cooled to -10 ° C in a cooling mixture of ice and sodium chloride. After the addition of 0.28 ml (2 mmol) of triethylamine, the solution becomes desalted and does not become liquid again by the addition of an additional 8 ml of dimethylformamide and vigorous stirring. 0.28 ml (2 mmol) of isobutyl chloroformate was added dropwise and after 30 minutes of activation time 0.16 g (2 mmol) of cystamine in 2 ml of dimethylformamide was added dropwise. The reaction mixture was stirred at -5 ° C for 2 hours, then at room temperature for 4 hours, then poured into a mixture of 50 ml of chloroform and 50 ml of petroleum ether. The resulting white precipitate is centrifuged, washed several times in a mixture of chloroform and petroleum ether, allowed to swell several times in alcohol and then precipitated with ether. 0.33 g of α-poly-γ-L-glutamylcystamine is obtained. Analysis: S 14.7%.

0.16 g of α-poly-γ-L-glutamylcystamine is suspended in 5 ml of glacial acetic acid and then 1 ml of 30% hydrogen peroxide is added. The reaction mixture was allowed to stand for 3 days at room temperature, whereupon the solution became clear. It is then diluted with water and lyophilized. 0.20 g of white .alpha.-poly-.beta.-L-glutamyltaurine is obtained, contaminated according to chromatographic analysis with taurine in an amount of 0.5%.

Analysis: S 11.9%.

The infrared spectrum of the product is identical to that described above.

Claims (1)

A process for the preparation of novel amino acid derivatives of the formula I, HaN — CH — CO — A ¹ I (CH ₂ ) " CO-N (CH _{2) t} ¹ -B R (I) W stands for A ^{1 is a} group of the general formula YNÁLEZU .................- ·· - (NH — CH — CO) _{r —} Y, R ³ in which Y represents a hydroxyl radical or a (C 1 -C 4) alkoxy radical, r denotes an average degree of polymerization of at most 2000, R ³ represents a group of the formula - (CH 2) -CO-N- (CH 2) t -B ¹ in which B ¹ represents a group of the formula —SO2 OH, —OSO2 OH or - OPO (0H) 2, R is a hydrogen atom or an alkyl radical having 1 to 4 carbon atoms, n an integer from 1 to 4 and an integer from 2 to 6, as well as their pharmaceutically acceptable salts or their optically active antipodes, characterized in that the compound of the formula II, H2N — CH — CO — A ³ (CH2 Cl2) CO — A ² (II) wherein n is as defined above, A ³ is - (NH-CH-CO) _r -Y, R ⁴ in which Y and r are as defined above and R ⁴ is - (CH ² ) n - CO - A ² , wherein n is as defined above and A ² is p-nitrophenoxy, pentachlorophenoxy or C 2 -C 4 alkoxycarbonyloxy, reacted with a compound of formula III, HN - (CH ^{2 -} - B ² R (III) wherein R and t are as defined above and B ² represents a group of the formula --SO ² OH. OSOaOH or OPO (OH) 2 or mercapto, and when B ² is mercapto, the resulting compound is oxidized and optionally converted to a salt, released from the salt and / or prepared in optically active form using optically active starting materials or by resolution of the racemic mixture formed as the product.