JPS6124377B2 - - Google Patents

Description

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

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

[wherein A ¹ is aralkoxy (the alkyl residue in the aralkoxy is lower alkyl, and the aryl residue may be substituted with methoxy or nitro) or the general formula -NHCH ₂ COY (where Y
is hydroxy or lower alkoxy), R ¹ is aralkoxycarbonyl (the alkyl residue in the aralkoxycarbonyl is lower alkyl, and the aryl residue may be substituted with methoxy or nitro) and B ¹ is the formula −SO ₂ OH, −OPO(OH) ₂ or −S−
SR ³ (R ³ is a residue obtained by removing the group B ¹ from the general formula ()), R represents hydrogen or lower alkyl, R ² represents hydrogen or carboxy, n is 1 or 2, and t is 1 or 2] 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. For example, the special public interest public in 1983
Compounds obtained by removing the protecting groups or substituents R ¹ and A ¹ on the α-amino group and α-carboxy group of some of the compounds of the present invention () by the method of No. 12347 In particular, the following formula () gamma-L-glutamine taurine has advantageous biological activities.

This compound is called “AGAS” (Aerobiospherical-Genetical-
It has a wide range of therapeutic and preventive effects against pathological changes directly or indirectly related to damage to the adaptive system.

To explain the concept of AGAS, 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 cavity and pharyngeal cavity, respiratory tract, and lungs); (b) Skeletal system and body Limbs (tubular and cancellous bones, ball and socket joints, synovium, skeletal musculature); (c) organs involved in the regulation of terrestrial ionic balance (transepithelial transport systems, intestinal villi and renal tubules); ( d) cisternal bacteria (associated with the tooth base and anchored by the tooth root) necessary for the decomposition of solid 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.

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

Significantly increased stress effects on terrestrial life (e.g.
meteorological and severe diurnal changes, increased risk of damage)
In contrast, this compound tends to stabilize the adaptation syndrome by simultaneously preventing peripheral tissue damage (eg, damage to connective tissue, bone matrix, etc.) induced by glycocorticosteroids.

Development of immune dynamic equilibrium (improved recognition of self and non-self cells).

The compounds of formula () exert their activity partly directly and partly through inhibition of vitamin A metabolism through the production of more polar vitamin A metabolites. This activity is associated with renal canalicular 25-hydroxy-cholecalciferol-1-α-hydroxylase (25-hydroxy-cholecalciferol-1-α-hydroxylase).
-hydroxylase) enzymes similar to those caused by parathyroid hormones. The above facts explain the wide and diverse biochemical, pharmacological and therapeutic activities of this compound.

(A) Effects of vitamin A properties (a) Pharmacological and biochemical effects Action to promote the incorporation of labeled sulfate into rat cartilage and into the lens, liver and lung tissues of chicken embryos; rat cartilage Action to promote the incorporation of labeled phosphorus into the body; action to promote the synthesis of chondroitin sulfate; action to favor wound healing (also effective against the reduction in wound healing induced by administering cortisone to rats and dogs). (effective); 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; traces Actions that affect the alternation of sexual elements (silicon, copper, zinc, manganese, fluorine); Actions that promote epithelial formation; Actions that increase the activity of alkaline phosphate enzymes; Induced by local action of vitamin A Action exerted on cyst production; very flat run of the dose-response curve and changes in aura symptoms at high doses; action to activate the Golgi apparatus; action to promote the production of goblet cells; serum Action to increase the concentration of vitamin A.

(b) Use in clinical treatment Keratoconjunctivitis sicca; Schjögren's syndrome; nasopharyngitis sicca; nasal eschar; chronic bronchitis; synobronchitis; pancreatic fibrosis; childhood pheumopathy tendency; periodontal increased predisposition of the skin and mucous membranes to viral and bacterial infections; antagonism of cortisone; surgical wounds and lesions of the mucous membranes; colonic erosions; pruritus; disorders of taste and smell.

(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; radioprotective action; Action to promote target reaching in maze test in active animals; Action to reduce experimental fluorosis and cadmium poisoning; Action to reduce experimental Egyptian bean toxicity; Renal cyclic adenosine-phosphate excretion Action to increase enzyme activity of liver tyrosine aminotransferase.

(b) Use in clinical treatment Less severe irradiation injury; vitiligo; asthenia; mental uplifting effect; effect of improving degenerative aging conditions and memory function; keloid predisposition; ankylosing spondylosis; disease of motor organs resulting from impairment ; sclerotic fundus; starch; patchy scleroderma; fibrocystic mastopathy.

The duration of treatment with this compound varies within wide limits. When given an oral dose of 5 μg of the chemically pure active substance three times a day, some patients no longer had symptoms after 2 weeks (e.g. nasopharyngitis sicca) and some other illnesses For some diseases, a treatment period of 1 to 2 months is required (eg, periodontal disease, Schjögren's syndrome), and for other diseases, a treatment period of 3 to 6 months is required (eg, ankylosing spondylosis).

The compounds of formula () can be made into cosmetic or pharmaceutical compositions for use in the treatment of humans and animals. The composition may contain only this compound as active ingredient or may contain other biologically active substances at any time. The active agents of the invention are preferably administered in doses of 50 to 500 nanograms per kilogram of body weight 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.) 2 to 20μ of the active ingredient when mixed with
g, preferably about 10 μg. 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 ingredient 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 μg.

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 μg 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 can be made with cocoa butter or synthetic waxes (e.g. imhausen mass,
GFR) with an active ingredient content of 2 to 20 μg, preferably about 10 μg.

Dermatological or cosmetic ointments prepared with common hydrophilic or hydrophobic ointment bases (e.g. cholesterol, paraffin, glycerin, lanolin, linseed oil, etc.) have an active ingredient content of 0.1 to 1.0μ
g/g is sufficient.

Aerosol formulations contain active ingredients at 0.1 to 1.0μ
It is preferable to contain it at a g/g concentration. The active ingredient content of sublingual tablets is approximately 10 μg 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 to 5 μg/g.
It can be in the form of a suspension. Similarly, long-acting injectable preparations can be prepared from mixtures of this polymer or a salt of a compound of formula () with high molecular weight organic bases (eg, protamine, histones). This composition contains 10 to 20 per ampoule.
Contains an amount of active ingredient in μg.

Dermatological and cosmetic powders may have an active ingredient content of 0.1 to 1 μg/g and contain the usual carriers (eg talc).

Eye drops and tear-miscible or immiscible ointments for ophthalmological applications have an active ingredient content of 0.1 to 1.0 μg/g.

The most preferred dosage for pediatric use is
The ratio is 0.3 μg of active ingredient per 1 kg of body weight.

Preferably, all disinfectant compositions are prepared by sterilization.

The combination of the above formulations containing the compound of the present invention increases the intended prophylactic, therapeutic or cosmetic effect;
to strengthen or improve; Primarily, the following co-supplementary ingredients will be used: vitamin A, vitamin C,
vitamin E, vitamin K, trace elements, cortisone and its derivatives, progesterone, thyroid hormones, products of radium-like and immunosuppressive action, psychotropic drugs (especially tranquilizers and thymoleptics), 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 formula () 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 improve vitamin A absorption and metabolism. This compound improves the absorption of trace elements,
It also increases its blood level. When used as a feed additive, it can be administered to animals in daily oral doses of 100 to 300 nanograms per kilogram of body weight, preferably about 200 nanograms. this is,
When mixed with animal feed, generally 1 kg feed
concentrations of 1 to 2 μg (i.e. 1 to 2 mg/
Considering that the required concentration is very low, corresponding to 0.001 to 0.002 ppm), the formula (
) can also be incorporated into microcapsules containing vitamin premixes or other useful feed additives, and can also be administered as an additive to drinking water or licking salts. The compounds can also be used in veterinary medicine in the same form as in human treatment (epithelialization, wound healing, bone fractures, etc.).

A common structural feature of compounds of general formula () is that they contain an α-substituted dicarboxylic acid moiety, and their ω
- the carboxyl group is bonded via an amide bond to a primary or secondary amino group which contains a strongly acidic group in the ω position in addition to other substituents in the alkyl side chain; be.

A compound of general formula () or a salt thereof can be produced according to the present invention by the following method.

General formula () [In the formula, R ¹ , A ¹ and n are each as defined above, and A ² is hydroxy, p-nitrophenoxy or pentachlorophenoxy] A compound of the general formula () [In the formula, R, R ² and t are each as defined above; B ² is a formula -SO ₂ OH, -OPO(OH) ₂ or -S-
S--R ⁴ (R ⁴ is the residue obtained by removing the group B ² from the general formula ())] and, if desired, the obtained compound is converted into its salt.

According to the method of the invention, compounds of general formula () are prepared by forming an acid amide bond. In this case, a suitable α-amino group and a carboxyl group having a protecting group or another substituent may be used.
-aminodicarboxylic acid derivatives via their ω-carboxyl group, such as 2-aminoethanesulfonic acid, 3-aminopropanesulfonic acid, 2-
Coupling with phosphoethanolamine, 3-phosphopropanolamine or cysteic acid. A wide range of protecting groups can be applied in this reaction. The most preferred method of attachment is the "active ester" method. For information on the generation and selective removal of protecting groups, as well as the bonding method, please refer to
E. Schrodor and K. Lubke, “The Peptides”
Vol.1, Peptide Synthesis Method, Academic Press
(1965).

It can also be achieved, for example, by acylating the amino group of distamine or a substituted cystamine derivative with an α-aminodicarboxylic acid derivative. Several coupling methods can be used to effect the acylation, such as the "active ester" or "mixed anhydride" method. When the product of this bonding reaction is reacted with hydrogen peroxide or peracid, another compound of the general formula () is produced through oxidative elimination of the disulfide bond.

The present invention will be explained in more detail by the following examples. However, the present invention is not limited to these.

Example 1 40.85 g (0.11 mol) of carbobenzyloxy-L-glutamic acid α-benzyl ester (Liebig's Ann. 655, 200/1962) Dissolve in 500ml of acetonitrile. Cool the solution to −15°C by excluding atmospheric moisture and add 15.4 ml.
(0.11 mol) of triethylamine is added to the stirred solution followed by 15.4 ml (0.11 mol) of isobutyl chloroformate. This mixture was heated at −15°C.
_{Stir for 40 min at} Add 250ml of acetonitrile. The mixture is stirred vigorously for 2 hours at -15°C and then for 4 hours at room temperature.

The reaction mixture is evaporated in vacuo at 30°C. The residue is mixed with 200 ml of ice-cold water under cooling and stirring, and the mixture obtained is evaporated in vacuo at 35°C. 250ml for residue
of water and 500 ml of ethyl acetate and place the mixture in a separating funnel. 250ml of ethyl acetate phase
of water, 2 x 250ml of 5% aqueous sodium carbonate solution,
Wash with 2 x 250 ml of 1N hydrochloric acid and 250 ml of water. (The aqueous-alkaline washing solution is acidified with hydrochloric acid,
After extraction with ether, about 5 g of unreacted carbobenzyloxy-L-glutamic acid α-benzyl ester is recovered. ) Dry the ethyl acetate solution over anhydrous sodium sulfate and evaporate to dryness under vacuum at 30 °C. A thick oily residue is obtained which crystallizes on standing. The residue is triturated with 250 ml of anhydrous ether, the crystalline material is separated off, and about 40-42 g of the crude product thus obtained is recrystallized from 100 ml of ethyl acetate and 170 ml of ether. 29.3 g N,N'-bis[N-carbobenzyloxy-gamma-(α-benzyl)-L-glutamine]-cysteamine is obtained. mp91−92℃.

Analysis C ₄₄ H ₅₀ N ₄ O ₁₀ S ₂ (MW=859.05) Calculated values: C 61.52%, H 5.89%, N 6.52%, S 7.46% Actual values: C 60.85%, H 5.91%, N 6.61%, S 7.72% Example 2 5.42 g (11 mmol) of carbobenzyloxy-L-glutamic acid (α-benzyl)-γ-p-
Nitrophenyl ester (Chem.Ber.96, 204/19
63) Dissolve in 50 ml of pyridine. The solution is cooled to 0<0>C and a solution of 1.25 g (10 mmol) of taurine _NH2- ( _CH2 ) ₂ - _SO2OH in 20 ml of water is added dropwise over 30 minutes under vigorous stirring. Then 3.08 ml (22 mmol) of triethylamine are added dropwise to the mixture and cooling and stirring are stopped. The mixture is left at room temperature for 72 hours and then evaporated in vacuo. Dissolve the residue in 50 ml of water and add 1N hydrochloric acid to the solution until the yellow coloration disappears. The solution is washed with 10 x 50 ml of ether to remove p-nitrophenol and the aqueous phase is evaporated in vacuo. 6.9g carbobenzyloxy
γ-(α-benzyl)-L-glutamyltaurine The triethylammonium salt of is obtained.

Example 3 0.47 g (1 mmol) carbobenzyloxy-α-L-glutamyl-(γ-p-nitrophenyl ester)-glycine methyl ester Dissolve in 6 ml of pyridine. This solution was cooled in an ice bath, and 0.125 g (1 mmol) of taurine was added to it for 2 hours.
ml of the solution in water and then 0.28 ml (2 mmol) of triethylamine are added. Each reactant should be added in small portions to always obtain a clear solution. The reaction mixture is left at room temperature for 3 days and then evaporated in vacuo. The oily residue is triturated with ether and petroleum ether and dried over sulfuric acid under vacuum. Carbobenzyloxy-α-L-glutamyl-(γ-taurine)-glycine methyl ester is obtained.

Example 4 1.083 g (2.2 mmol) of carbobenzyloxy-L-glutamic acid (α-benzyl)-γ-p
- The nitrophenyl ester is dissolved in 6 ml of a 2:1 pyridine-water mixture and 278 mg (2 mmol) of homotaurine NH ₂ -(CH ₂ ) ₃ -SO ₂ OH and 0.59 ml.
(4.2 mmol) of triethylamine is added to this solution. The resulting yellow solution is left at room temperature for 72 hours and then evaporated in vacuo. The oily residue is dissolved in water, neutralized with hydrochloric acid and subjected to ether extraction in a continuous extractor for 8 hours to remove p-nitrophenol. Evaporation of the aqueous phase in vacuo yielded 1.68 g of carbobenzyloxy-γ-(α-benzyl)-L-glutamyl-homotaurine. is obtained.

Example 5 1.083 g (2.2 mmol) of carbobenzyloxy-L-glutamic acid (α-benzyl)-γ-p
- React the nitrophenyl ester with 278 ml (2 mmol) of N-methyltaurine in the manner described in Example 4. 1.59g carbobenzyloxy-γ
-(α-benzyl)-L-glutamyl-N-methyltaurine is obtained.

Example 6 2.87 g (6.6 mmol) of carbobenzyloxy-L-glutamic acid (α-benzyl)-γ-p
- Dissolve the nitrophenyl ester in 20 ml of pyridine and add 1.25 g (6 mmol) of L-cysteic acid. Add a solution of the hydrate in a mixture of 17 ml of water and 17 ml of pyridine. 2.6ml of this mixture
(18.6 mmol) of triethylamine is added and the reaction mixture is left at room temperature for 72 hours. Add this solution to 30
Evaporate in vacuo at °C. The residue is dissolved in 20 ml of water and the solution is acidified with concentrated hydrochloric acid and then washed with 15×10 ml of ether. The aqueous phase is evaporated in vacuo at 35°C.
Carbobenzyloxy-γ-(α-benzyl)-L
-glutamyl-cysteic acid is obtained.

Example 7 1.083 g (2.2 mmol) of carbobenzyloxy-L-glutamic acid (α-benzyl)-γ-p
-Nitrophenyl ester in pyridine and water:
282 mg (2 mmol) of colamine phosphate (U.S. Pat.
2730542) Add _NH2- ( _CH2 ) ₂ -OPO(OH) ₂ and 0.87 ml (6.2 mmol) of triethylamine. The mixture is left at room temperature for 72 hours and then evaporated in vacuo. The residue is treated as described in Example 4. 1.25g carbobenzyloxy-γ-
(α-benzyl)-L-glutamyl-colamine phosphate is obtained.

Example 8 526 mg (1.1 mmol) of carbobenzyloxy-L-aspartic acid (α-benzyl)-β-p
-Nitrophenyl ester (Chem.Ber.97,
1789/1964) Dissolve in 5 ml of pyridine. This solution was cooled to 0°C, and a solution of 125 mg (1 mmol) of taurine dissolved in 2 ml of water was added little by little.
Add ml (2 mmol) of triethylamine.
The reaction mixture is left at room temperature for 48 hours and then evaporated in vacuo. The residue is dissolved in 5 ml of water and 1N hydrochloric acid is added dropwise to this solution until the yellow coloration disappears.
10% solution to remove p-nitrophenol
Wash with x5 ml of ether and evaporate the aqueous phase in vacuo. 478 mg of carbobenzyloxy-β-(α-benzyl)-L-aspartyltaurine is obtained.

Example 9 526 mg (1.1 mmol) of carbobenzyloxy-L-aspartic acid (α-benzyl)-β-p
- When the nitrophenyl ester is reacted with 139 mg (1 mmol) of homotaurine as in Example 8, carbobenzyloxy-β-(α-benzyl)-L-aspartylhomotaurine is obtained.

Example 10 Carbobenzyloxy-L-aspartic acid.
(α-benzyl)-β-p-nitrophenyl ester is reacted with colamine phosphate as in Example 7. Carbobenzyloxy-β-(α
-benzyloxy)-L-aspartyl cholamine phosphate is obtained.

Claims (1)

[Claims] 1 General formula () [wherein A ¹ is aralkoxy (the alkyl residue in the aralkoxy is lower alkyl, and the aryl residue may be substituted with methoxy or nitro) or the general formula -NHCH ₂ COY (where Y
is hydroxy or lower alkoxy), R ¹ is aralkoxycarbonyl (the alkyl residue in the aralkoxycarbonyl is lower alkyl, and the aryl residue may be substituted with methoxy or nitro) and B ¹ is the formula −SO ₂ OH, −OPO(OH) ₂ or −S−
SR ³ (R ³ is a residue obtained by removing the group B ¹ from the general formula ()), R represents hydrogen or lower alkyl, R ² represents hydrogen or carboxy, n is 1 or 2, and t is 1 or 2] In producing the compound or its salt, the general formula () A compound of [wherein R ¹ , A ¹ and n are each as defined above, and A ² is hydroxy, p-nitrophenoxy or pentachlorophenoxy] is represented by the general formula () [In the formula, R, R ² and t are each as defined above; B ² is a formula -SO ₂ OH, -OPO(OH) ₂ or -S-
S-R ⁴ (R ⁴ is the residue obtained by removing the group B ² from the general formula ())] and, if desired, converting the obtained compound into its salt. How to become.