JPS6049629B2 - Production method of new amino acid derivatives - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel amino acid derivative and a method for producing the same.

The novel compound of the present invention corresponds to the following general formula (1).

In the formula, A1 is hydroxy, C1-4 alkoxy, or phenyl-C1-4 alkoxy, which may be substituted with methoxy or nitro, or the general formula, -NHCFI2C
OY (Y is hydroxy or C1-4 alkoxy), R represents hydrogen or methyl, R1 is hydrogen, C1-4-alkoxycarbonyl or C
Represents phenyl-C1-3 alkoxycarbonyl which may be substituted with 1-4 alkoxy or nitro,
n is 1 or 2 and t is 2 or 3.

Salts and optically active isomers of the above compounds are also included within the scope of the present invention.

Some of the novel compounds of the invention have useful pharmaceutical properties, while others can be used as intermediates in the preparation of compounds with useful physiological or pharmaceutical properties.

With respect to physiological activity, outstandingly advantageous among the new compounds of the invention are gamma L-glutamyltaurines of the general formula.

This compound is called “AGAS” (AerObiOspherical-Genetica).
It has a wide range of therapeutic and preventive effects against pathological changes directly or indirectly related to damage to the I-Adaptation Onal-System.

To explain the AGAS (7) concept, we will list the most important tissues and organs that make up this system.

(a) Biological interfaces that form the boundary between living organisms and the atmosphere as their habitat (skin and other skin-like structures, cornea and conjunctiva, oral and pharyngeal cavities, airways, and lungs) (b) Skeletal system and body limbs (tubular and cancellous bones, ball and socket joints, synovium, skeletal muscle tissue) (c) Terrestrial organs involved in the regulation of ionic balance (transport system through the epithelium, intestinal villi and renal tubules) (d) Solid alveolar teeth (with tooth bases and fixed by the roots) necessary for the breakdown of food; (e) terrestrial auditory, olfactory and sound-forming organs;

The compounds prepared according to the invention exert biologically favorable therapeutic effects on the organs of the above-mentioned systems as well as their tissues.

Moreover, regarding the AGAS system, the compounds of the present invention also exhibit the following effects: radiation protection, wound healing promotion, systemic mensenchyma
) activation effect, protection against increased risk of infection and contamination of the mucous membranes and skin (lysozyme production of moist mucous membranes, development of the hairy epithelium of the respiratory tract, etc.), protection against viral and bacterial infections of the skin. Improved.

In response to the significantly increased stress effects of terrestrial life (e.g., climatic and strenuous, peropharyngeal changes, increased risk of injury), this compound has been shown to reduce peripheral tissue damage induced by glycocorticosteroids ( It tends to stabilize the adaptation syndrome by simultaneously preventing damage to the connective tissue bone matrix, etc.).

Development of immunodynamic equilibrium (increased recognition of self and non-self cells) The compounds of the invention inhibit the effects of vitamin A metabolism, partly directly and partly through the production of more polar vitamin A metabolites. It shows its activity through.

This activity is associated with renal canalicular 25-hydroxycholecalciferol-1-α-hydroxylase (25-HydrO
xy-ChOlecalciferOl-1-α-Hy
drOxylase) enzyme is similar to that caused by parathyroid hormones. The above facts explain the wide and diverse biochemical, pharmacological and therapeutic activities of the compounds of the present invention. Effects of vitamin A properties (a) Pharmacological and biochemical effects Action to promote the incorporation of labeled sulfate into rat cartilage, and into the crystalline lens, liver and lung tissues of chicken embryos; into rat cartilage Action to promote the incorporation of labeled phosphorus; action to promote the synthesis of chondroitin sulfate; action to favor wound healing (also effective in reducing wound healing induced by administering cortisone to rats and dogs). ); action to increase granule reduction in mast cells; action to enhance vitamin A in cases of experimental vitamin deficiency or hyperemia in rats and chickens; action to reduce stress ulcers in rats, action to increase lysozyme production; trace elements (silicon, copper, zinc, manganese, fluorine): Effects that promote epithelial formation: Effects that increase alkalinity and phosphate enzyme activity; Capsule formation induced by the local action of vitamin A Very flat dose-response curve and changes in aura symptoms at high doses: Activating the Golgi apparatus; Promoting the production of goblet cells; Serum vitamin A The effect of increasing the concentration of.

(b) Use in clinical treatment
pancreatic fibrosis; childhood tendency towards pheumopathy; periodontitis; increased predisposition of the skin and mucous membranes to viral and bacterial infections; cortisone anti-inflammatory effects; surgical wounds and lesions of the mucous membranes; colonic erosions; Pruritus group; taste and... Anosmia.

(B) Effects of non-vitamin A properties (a) Pharmacological and biochemical effects Transient hypoglycemic action; action to reduce phosphate urine and increase serum phosphate levels; radiation. Radioprotective action; Action to promote target reaching in maze test in inactive animals; Action to reduce experimental fluorosis and cadmium poisoning; Action to reduce experimental Egyptian bean toxicity; Renal cyclic adenosine - Action to increase phosphate excretion; action to increase enzyme activity of hepatic tyrosine aminotransferase.

(b) Use in clinical treatment Less severe irradiation injuries: vitiligo: myasthenia; mental uplifting effect; effect to improve degenerative aging condition and memory function; keloid predisposition; ankylosis formation and spondylosis; motor organ tract derived from impairment disease; sclerotic basis (ScIerOticfurldus)
; starchyosis; patchy scleroderma; fibrocystic mastopathy.

The duration of treatment with the compounds of the invention varies within wide limits.

When given the chemically pure active substance at an oral dose of 5 μDa three times a day, some patients no longer had symptoms after 2 weeks (e.g., sicca rhinolaryngopharyngitis); Treatment for some diseases may require 1 to 2 months (e.g. periodontal disease, Schjögren's syndrome), and for other diseases up to 3 months.
A treatment period of up to 6 months is required (eg, ankylosis formation or spondylosis). The compounds of the invention can be converted into cosmetic or pharmaceutical compositions for use in the treatment of human resources.

The compositions may contain only the compounds of the invention as active ingredients, or may contain other biologically active substances at any time. The active agent of the invention weighs 1 kg.
Preferably, the dosage is 50 to 500 nanograms per gram given three times a day. One tablet consists of a biologically inert carrier (e.g. lactose, starch) and the usual auxiliary substances (e.g. granulating agents and lubricants such as polyvinylpyrrolidone, gelatin, talc, magnesium stearate, ultrafine silica, etc.) The active ingredient of the present invention in admixture with 2 to 2
0μy1 preferably contains about 10py.

Considering this very low dosage, in order to homogeneously disperse this active substance in the tablet, the active ingredient in solution form is mixed with the tablet mass before granulation and a homogeneous mixture is created using a kneader. Preferably, it is prepared. Because the required effective dosage is so low, the active ingredients of the invention can be produced at a satisfactory cost in large laboratory-scale equipment, even when producing trillions of tablets. The active ingredient is stable, so the tablets can be stored for long periods of time. The active ingredient content in depot tablets or Spansuled capsules is 10 to 30 μf. Injectable preparations containing the active ingredient of the invention in powder ampoules, optionally mixed with a biologically inert water-soluble diluent, contain from 5 to 10 μq of active ingredient per ampoule. It is preferable to be there.

Parenteral application is preferably by intramuscular, subcutaneous or intravenous injection. Since the active ingredients of the invention at a given concentration do not irritate tissues or vessel walls, they can also be applied in the form of drops. Suppositories are prepared using cocoa butter or synthetic waxes (e.g. Imhausen, Mass, GFR) that can be used for this purpose.
It can be prepared with an active ingredient content of from 20 μy1, preferably about 10py.

Common hydrophilic or hydrophobic ointment bases (e.g. cholesterol, paraffin, glycerin, lanolin, linseed oil,
The dermatological or cosmetic ointments prepared in the above may have an active ingredient content of 0.1 to 1.0 μYly. The aerosol formulation preferably contains the active ingredient in a concentration of 0.1 to 1.0 μYly.

The active ingredient content of sublingual tablets is about 10 py per tablet, and the disintegration time is 0.5 to 1 hour. Polymers of high molecular weight with a sustained effect can also be prepared, for example with an active ingredient content of 1
It can be in the form of a suspension of 5μYlq(7).

Similarly, long-acting injectable preparations can be prepared from mixtures of the polymers or salts of the compounds of the invention with high molecular weight organic bases (eg, protamine, histones). The composition contains active ingredient in an amount of 10 to 20 μq per amble. Dermatological and cosmetic powders may have an active ingredient content of 0.1 to 1 μm and contain the usual carriers (eg talc).

Eye drops applied for ophthalmology and ointments miscible or immiscible with tears have an active ingredient content of 0.1 to 1.0 p.
It is y1y.

The most preferable dose for pediatric use is 1 kg body weight.
The ratio is 0.3 μq of active ingredient per g.

Preferably, all sterile compositions are prepared by sterile filtration. Combinations of the above formulations containing the compounds of the invention increase, intensify or improve the intended prophylactic, therapeutic or cosmetic effect.

Primarily, the following co-supplementary ingredients will be used: Vitamin A
1 vitamin C1 vitamin E1 vitamin K1 trace elements,
Cortisone and its derivatives, progesterone, thyroid hormones, products of radium-like and immunosuppressive action, psychotropic drugs (especially tranquilizers and thymoleptics, Thyr
TlOleptics), organosilicon compounds, gerontological preparations, oral antidiabetic agents, anti-inflammatory agents, antihistamines, etc. The appropriate amount of each component in the combination formulation is generally about the same as the usual therapeutic amount when used alone. The compounds of the invention can also be used as additives in therapeutic and nutritional premixes.

When used in such compositions, the compounds increase weight gain and also reduce vitamin A requirements and/or
Or improve vitamin A absorption and metabolism. This compound improves the absorption of trace elements and also increases their blood levels. When used as a feed additive, it weighs 1k
100 to 300 nanograms per 9, preferably about 2
A daily oral dose of 0.00 nanograms can be administered to animals. When mixed with animal feed, this is generally at a concentration of 1 to 2 μy/k9 feed (i.e. 1 to 2 m9/tonne or 0.001 to 0.002 ppm).
). In view of the very low concentrations required, the compounds of the invention can also be incorporated into microcapsules containing vitamin premixes or other useful feed additives, or added to drinking water or licking salts. It can also be administered as a drug. The compounds of the invention can also be used in veterinary medicine in the same manner as they are applied in human treatment (epithelialization, wound healing, bone fractures, etc.). General formula (1
The common structural feature of compounds of It is bonded to a primary or secondary amino group via an amide bond.

The compound of general formula (1) or its salt or optically active isomer can be produced according to the present invention by the following method.

Formula (■) [In the formula, A1, R, . Rl, n and t are each as described above, and B is halogen, hydroxy, p
-toluenesulfonyloxy, or a group of formula -SHI or -S-S-R2 (R2 is a residue when group B is removed from formula (■))], in which group B
is converted to the group -SO2OH.

More specifically, according to the present invention, general formula (■A) [wherein N. ．． R. ．． Rl, n and t have the same meanings as above,
and B1 is of the formula -SH or -S-S-R2 (in the formula R2
is the residue obtained when group B1 is removed from general formula (■A)], or the compound of general formula (■B) [in which A1, R, . Rl, n and t are as defined above] by reaction with p-toluenesulfonic acid chloride and subsequent treatment with an alkali metal sulfite or bisulfite to form a SO2OH group. or the general formula [wherein A1, R. ．． Rl, n and t are as defined above and B2 is halogen or p-toluenesulfonyloxy] with an alkali metal sulfite or bisulfite, if desired, the resulting product α - on the amino group and/or α
The compound of general formula (1) or a salt thereof can be produced by a method comprising removing the protecting group on one carboxy group and, if necessary, converting the obtained compound into a salt or liberating it.

When the starting material has a disulfide bond, when it is reacted with hydrogen peroxide or peracid, the compound of general formula (1) is produced through oxidative cleavage of the disulfide bond. Removal of the protecting group can be carried out at any stage.

For example, when the -0H group of the compound of general formula (■B) is converted to -SO2OH group by reaction with p-toluenesulfonic acid chloride and subsequent treatment with an alkali metal sulfite or bisulfite. The removal of the protecting group can be carried out before or after treatment with an alkali metal sulfite or bisulfite. The present invention will be explained in more detail by the following examples.

However, the present invention is not limited to these. 7
Example 1 40.85y (0.11 mol) of carbobenzyloxy-L-glutamic acid α-benzyl ester (Lieb
ig3sAnn. ? ? , 200/1962) for 500m
Dissolve in t acetonitrile.

The solution was cooled to -150°C by excluding the moisture in the air, and the temperature was 15.4T! Ll (0.11 mol) of triethylamine was added to the stirred solution, followed by 15.4 ml (0.1 mol) of triethylamine.
．． 11 mol) of isobutyl chloroformate are added. This mixture was stirred at -15°C for 4 minutes, and then 28ml (a). 2 mol) of triethylamine and 11.26y (0.
05 mol) of cysteamine dihydrochloride and finally 250 m
Add L of acetonitrile. This mixture was heated to -15°C.
Stir vigorously for 2 hours at room temperature and then for 4 hours at room temperature. The reaction mixture is evaporated in vacuo at 30'C.

The residue was mixed with 200 ml of ice-cold water under cooling and stirring,
The resulting mixture is evaporated in vacuo at 35°C. 250 for residue
ml of water and 500 ml of ethyl acetate are added and the mixture is placed in a separate load. 250ml of ethyl acetate phase
of water, 2 x 250 ml of 5% aqueous sodium carbonate solution, 2
Wash with 250 ml of 1N hydrochloric acid and 250 ml of water. (The aqueous-alkaline wash is acidified with hydrochloric acid and extracted with ether to recover approximately 5 Da of unreacted carbobenzyloxy-L-glutamizoic acid α-benzyl ester.) The ethyl acetate solution is dried over anhydrous sodium sulfate. death,
Evaporate to dryness under vacuum at 30°C. A thick oily residue is obtained which crystallizes on standing. The residue was triturated with 250 ml of anhydrous ether, the crystalline material was filtered off and the roughly 40-42y crude product thus obtained was dissolved in 100 ml of anhydrous ether.
of ethyl acetate and 170 mt of ether.
29.3 V of N-N''-bis[N-carbobenzyloxy-gamma(α-benzyl)-glutamyl]-cystamine is obtained.

M. p. 9l~97C0 analysis C4, H5ON4OlOS
2 (MW=859.05) Calculated value: C6l. 52%, H
5.89%, N6.52%, S7.46% Actual value: C6
O. 85%, H5.9l%, N6.6l%, S7.72
%25.77y (0.03 mol) of N-N''-bis[
N-carbobenzyloxy-γ-(α-benzyl)-
[glutamyl]-cystamine is dissolved in 75 mt glacial acetic acid.

The solution is cooled in an ice bath and a freshly prepared mixture of 75 ml of 30% hydrogen peroxide and 225 ml of glacial acetic acid is added dropwise in one drop. The ice bath is then removed and the mixture is stirred at room temperature for 4 hours and evaporated in vacuo at 30°C. The oily product is then placed in a desiccator and dried first over phosphorus pentoxide and then over solid potassium hydroxide. 28.5y of carbobenzyloxy-γ-(α-benzyl)-glutamyltaurine is obtained.

This crude product can be used for the production of γ-L-glutamyl taurine without purification. Example 20.48y (1 mmol) of carbobenzyloxy-alpha-glutamyl(γ
-p-nitrophenyl ester)-glycine ethyl ester (ActaChlm.Acad.Scj.Hung
．． ? 375/1970) in 6 ml of ethyl acetate.

This solution is cooled to O'C with ice water, and a solution of 0.08y (1 mmol) of cysteamine dissolved in Lml of dimethylformamide is added. After that, 0.14m1 (1
Triethylamine (mmol) is added dropwise to this solution. The precipitate gradually begins to separate. The reaction mixture is left in ice water and then at room temperature for 1 day. The mixture is diluted with a 1:1 mixture of ethyl acetate and ether and the precipitate is centrifuged and washed several times with a 4:1 mixture of ether and ethyl acetate and finally once with ether. The precipitate is dried over sulfuric acid, then washed successively three times with IN hydrochloric acid, twice with water, twice with saturated aqueous sodium bicarbonate, and twice with water, and dried in vacuo over sulfuric acid. 0.35y (85%) of carbobenzyloxy-α-glutamyl(γ-cysteamine)-
Glycine ethyl ester is obtained.

Calculated as analysis Cl9H27O6N3S: C52.6%, H6. 1%, S7.8%, actual values: C53.4%, H6.5%, S7.7% IR Svectorini specific absorption maximum 3310 (NH), 1748 (ester carbonyl), 1690 (C = 0, carbobenzyloxy ) and 1655(amidecarbonyl)Cm-1.

100 mg of carbobenzyloxy-α-glutamyl(γ-cysteamine)-glycine ethyl ester
Dissolve in ml of glacial acetic acid and add 0.5 ml of 30% hydrogen peroxide to this solution. The reaction mixture is left in an ice bath for 4 hours. The progress of the reaction is monitored electrophoretically. Once the reaction is complete, the mixture is diluted with water and lyophilized. 0.1
A solid foamy carbobenzyloxy-α-glutamyl (γ-taurine)-glycine ethyl ester of 1y is obtained.

Yield 95%. Example 3 4.14y (10 mmol) of carbobenzyloxy-γ-(α-benzyl)-glutamylcolamine was added to 40m
1. Dissolve in pyridine.

The solution is cooled to -100 C and 2.1y (11 mmol) of p-toluenesulfonyl chloride is added in portions under vigorous stirring. The mixture was stirred at O'C for 3 hours,
Pour into 40y of ice. The precipitated material is filtered off, washed with water and recrystallized from a mixture of ethanol and petroleum ether. The obtained product was mixed with ethanol and water in a 4:1 (volume ratio)
Dissolve in the mixture, add 10% palladium-carbon carrier decomposition medium and catalytically hydrogenate the mixture under shaking. The dry material obtained after hydrogenation was dissolved in 30 mt of water and 10.1
y (40 mmol) of sodium sulfite heptahydrate are added and the solution is stirred at 40° C. for 24 hours and finally evaporated in vacuo. Dissolve the residue in a minimum amount of water and add this solution to the DOw
Pass through a column packed with ex5O ion exchanger and elute the column with water. The eluate is evaporated in vacuo and the residue is dried over potassium hydroxide. Recrystallization of the crude product from 80% ethanol yields 1.6y gamma supra-gretamyl urethane. Example 4 115 ml of thionyl bromide was mixed with 4.14 Da (10 mmol) of carbobenzyloxy-γ-(α-benzyl)-
Add glutamylcolamine and stir the mixture for 3 hours.

The mixture is then diluted with ether and the precipitate that has separated out is filtered off and recrystallized from a mixture of acetone and petroleum ether. The material obtained is dissolved in a mixture of dimethylformamide and water and 10.1y (40 mmol) of sodium sulfite heptahydrate are added in portions with stirring. The mixture is stirred for 24 hours at room temperature and then for 6 hours at 50° C. and finally filtered. The clear filtrate is evaporated in vacuo and the residue is catalytically hydrogenated as described in Example 3. The dry material obtained after hydrogenation is dissolved in a minimum amount of water and DOw
The solution is passed through a column packed with ex5O ion exchanger and the column is eluted with water. The eluate is evaporated in vacuo and the residue is recrystallized from 80% ethanol. 1.2q of gamma-glutamyl taurine is present.

Finally, preferred embodiments of the present invention will be summarized.

1 In a method of oxidizing a compound of general formula (■A),
A method of reacting with a mixture of peracid or glacial acetic acid and hydrogen peroxide.

2 A method of heating the compound of general formula (■C) in the presence of an alkali metal sulfite.

3 In preparing the compound of the present invention in the form of a salt, the general formula (1
) with an alkali metal or alkaline earth metal hydroxide or carbonate, or with an organic base.

Claims (1)

[Claims] 1 General formula (I) ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) [A^ in the formula
1 is hydroxy, C_1_-_4 alkoxy, or phenyl-C_1_-_4 alkoxy which may be substituted with methoxy or nitro, or the general formula -NHCH^
2COY (Y is hydroxy or C_1_-_4 alkoxy), R represents hydrogen or methyl, R^1 may be substituted with hydrogen, C_1_-_4-alkoxycarbonyl or C_1_-_4 alkoxy or nitro phenyl-C_1_-_3 alkoxycarbonyl, n is 1 or 2, and t is 2 or 3] A method for producing a compound of the general formula (IIA) ▲ mathematical formula, chemical formula, There are tables, etc. ▼ (IIA) [
In the formula, A^1, R, R^1, n and t have the same meanings as above, and B^1 represents the formula -SH or -S-S-R^2(
where R^2 is the residue obtained when the group B^1 is removed from the general formula (IIA)] and, if desired, the α-amino basic and/
Alternatively, a method comprising removing the protecting group on α-carboxy and, if necessary, converting the obtained compound into a salt or liberating it from the salt. 2 General formula (I) ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) [A^ in the formula
1 is hydroxy, C_1_-_4 alkoxy, or phenyl-C_1_-_4 alkoxy which may be substituted with methoxy or nitro, or the general formula -NHCH^
2COY (Y is hydroxy or C_1_-_4 alkoxy), R represents hydrogen or methyl, R^1 may be substituted with hydrogen, C_1_-_4-alkoxycarbonyl or C_1_-_4 alkoxy or nitro phenyl-C_1_-_3 alkoxycarbonyl, n is 1 or 2, and t is 2 or 3] A method for producing a compound of general formula (IIB) ▲ mathematical formula, chemical formula, There are tables, etc. ▼ (IIB) [
A compound of the formula A^1, R, R^1, n and t are as defined above] is reacted with p-toluenesulfonyl chloride and then treated with an alkali metal sulfite or bisulfite, if desired. Before or after the treatment, the protective groups on the α-amino group and/or α-carboxy of the product are removed and, if necessary, the resulting compound is converted into or released from the salt. How to become. 3 General formula (I) ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) [A^ in the formula
1 is hydroxy, C_1_-_4 alkoxy, or phenyl-C_1_-_4 alkoxy which may be substituted with methoxy or nitro, or the general formula -NHCH_
2COY (Y is hydroxy or C_1_-_4-alkoxy), R represents hydrogen or methyl, and R^1 is substituted with hydrogen, C_1_-_4-alkoxycarbonyl or C_1_-_4-alkoxy or nitro; phenyl-C_1_-_3 alkoxycarbonyl, n is 1 or 2, and t is 2 or 3] A method for producing a compound or a salt thereof of general formula (IIC)▲mathematical formula, chemical formula , tables, etc. ▼ [In the formula, A^1, R, R^1, n and t have the same meanings as above, and B^2 is halogen or p-toluenesulfonyloxy] is an alkali metal sulfite salt or bisulfite and, if desired, α of the resulting product.
- a process consisting of removing the protective group on the amino group and/or on the α-carboxylic group and, if necessary, converting the compound obtained into a salt or liberating it from the salt.