JPH11279041A - Elastase inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an elastase inhibitor comprising, as an active ingredient, a compound which has excellent elastase inhibitory activity, is easy to synthesize and has excellent safety. SOLUTION: The following general formula (1): (Where X is COOK, etc., Y is N (R ¹ ′), Z is C
ONH _{2 and the} like, R ¹ and R ¹ ′ represent a hydrogen atom and the like. An elastase inhibitor comprising a compound represented by the formula (1) or a salt thereof as an active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an elastase inhibitor which has an excellent elastase inhibitory activity and contains a compound which can be easily synthesized as an active ingredient.

[0002]

2. Description of the Related Art In recent years, studies on aging have been advanced, and the main factors that cause skin aging are the effects of aging, drying, oxidation, sunlight (ultraviolet rays) and the like. Skin aging is recognized by a decrease in collagen and elastin in the skin dermis, a decrease in mucopolysaccharides such as hyaluronic acid, damage to cells by ultraviolet rays, and the like.

[0003] Of these, elastin crosslinks each other and contributes to the elasticity of tissues. However, elastin denaturation and destruction due to overexpression of elastin, which is an elastin-degrading enzyme, due to exposure to ultraviolet light and aging, may cause skin damage. It is thought to lead to reduced elasticity. Therefore, suppressing the activity of elastase is important in terms of imparting elasticity and firmness to the skin and preventing aging of the skin. For this reason, substances that inhibit the activity of elastase have been studied so far, such as 1,10-orthophenanthroline and spurge (Euphorbiacea).
e) An extract of Menila (P. niruri L.) belonging to the genus Phyllanthus of the family (Japanese Patent Application Laid-Open No. 9-87136) is known as an elastase inhibitor.

[0004]

However, the above-mentioned extract of Meniran requires a long time for extraction and its effect is not always sufficient, and 1,10-orthophenanthroline also has an elastase inhibitory activity, but is not safe. Had problems.

Accordingly, an object of the present invention is to provide an elastase inhibitor which has an excellent elastase inhibitory activity and contains a compound which can be easily synthesized as an active ingredient.

[0006]

Means for Solving the Problems The present inventors have found that the compound represented by the general formula (1) has excellent elastase inhibitory activity and is easy to synthesize, and have completed the present invention. Was.

That is, the present invention provides the following general formula (1)
During,

[0008]

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Wherein X is COOR ² (wherein R ² represents a hydrogen atom or an alkyl group), and the following general formula (2)

[0010]

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(Wherein R ³ and R ⁴ represent a hydrogen atom, an alkyl group or an aralkyl group) and the following general formula (3)

[0012]

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(Wherein R^FiveIs a hydrogen atom, an alkyl group, or
Represents an aralkyl group;⁶Represents an aralkyl group. )
Or SO_ThreeH, Y is N (R¹') (Where R¹′
It represents a hydrogen atom or a lower alkyl group. ), O or CH_Two
And Z is CON (R⁷) R⁸ Or COOR⁹(Where
R⁷, R⁸And R⁹Represents a hydrogen atom or an alkyl group
You. ) And R¹Is a hydrogen atom, even if it has a substituent
Good alkyl, aralkyl or alkoxyl
Group or R¹And R¹′ Together form a ring
Is shown. )) Or a salt thereof as an active ingredient
And elastase inhibitors.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION In the general formula (1), R ¹ represents a hydrogen atom, an alkyl group which may have a substituent, an aralkyl group or an alkoxyl group. The alkyl group which may have a substituent refers to a straight-chain or branched-chain alkyl group having 1 to 12 carbon atoms or a substituent such as a carbamoyl group, a hydroxyl group, a carboxyl group, or the like having 1 to 6 carbon atoms. An alkyl group having the formula: Among them, a straight-chain or branched-chain alkyl group having 3 to 5 carbon atoms or an alkyl group having the above-mentioned substituent in the alkyl group having 1 to 4 carbon atoms is preferable, and among them, n-propyl group, isopropyl group and 2,2-dimethyl are preferable. A propyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, a carbamoylmethyl group, a 2-carbamoylethyl group, and a hydroxymethyl group are preferred. The aralkyl group is
A phenylalkyl group and a naphthylalkyl group are exemplified, of which a phenyl-C _1-6 alkyl group is preferable,
Benzyl group, phenethyl group, 1-naphthylmethyl group, 2
-A naphthylmethyl group is more preferred. Further, examples of the alkoxyl group include a linear or branched alkoxyl group having 1 to 12 carbon atoms.
Is preferably a linear or branched alkoxyl group.

In the general formula (1), X is represented by COOR ² , the general formula (2), the general formula (3) or SO ₃ H, preferably COOR ² . R ² is a hydrogen atom or an alkyl group, of which a hydrogen atom, a methyl group, an ethyl group,
A propyl group, an n-butyl group and an isobutyl group are particularly preferred. In the general formula (2), R ³ and R ⁴ are a hydrogen atom, an alkyl group or an aralkyl group, of which a hydrogen atom, a methyl group, an ethyl group and a benzyl group are preferred. In the general formula (3), R ⁵ is a hydrogen atom, an alkyl group or an aralkyl group, of which a hydrogen atom is preferable. R ⁶ is an aralkyl group, of which a benzyl group and a phenethyl group are preferred. Y is N (R ¹ '), O or CH ₂ ;
Preferably it is N (R ¹ '). R ¹ ′ represents a hydrogen atom, a lower alkyl group, or a combination of R ¹ and R ¹ ′ to form a ring. R ¹ ′ is a hydrogen atom or R ¹
And R ¹ 'preferably together form a ring. Z
Is CON (R ⁷ ) R ⁸ or COOR ⁹ , of which CON (R ⁷ ) R ⁸ is preferable. R ⁷ and R ⁸ are a hydrogen atom or an alkyl group, which may be the same or different, and are preferably both hydrogen atoms.

The salt of the compound represented by the general formula (1) includes an alkali metal salt, an alkaline earth metal salt, an amine salt, an amino acid salt and the like. Preferred are alkali metal salts and amino acid salts. The compound represented by the general formula (1) may have optical activity, and the configuration may be either R or S, or may be a racemic form. Further, the present compound may be in the form of a hydrate.

Specific examples of the compound represented by the general formula (1) include the following.

[0018]

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[0019]

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Of these, compounds 1, 2, 3, 5,
6, 7, 8, 13 and 14 are particularly preferred because they have excellent elastase inhibitory activity, are easy to synthesize and are excellent in safety.

The compound represented by the general formula (1) can be produced, for example, by the following method. When Y in the general formula (1) is N (R ¹ ′), for example, the compound is reacted with a hydrochloride of an amide compound and a halogenated ester, a halogenated phosphoric acid ester or the like in the presence of a base such as an amine, If necessary, a base such as potassium hydroxide is reacted, and the obtained reaction product is appropriately extracted, washed, dried and the like to obtain the desired product. When Y in the general formula (1) is O, for example, the corresponding alcohol derivative is reacted with phosphoryl chloride, chlorosulfuric acid, or the like, and the obtained reaction product is appropriately extracted, washed, dried, and the like to obtain the desired product. . When Y in the general formula (1) is CH ₂ , for example, an α-halogenocarboxylic acid derivative and a malonic acid diester derivative are reacted in the presence of a base such as potassium carbonate, and then a decarboxylation reaction is performed. To react with an acid anhydride such as acetic anhydride and react with an amine such as ammonia.
The obtained reaction product is appropriately extracted, washed, dried, and the like to obtain the desired product.

The amount of compound (1) to be added to the elastase inhibitory activity of the present invention is not particularly limited.
For example, 0.0001 to 40% by weight is preferable,
1-20% by weight is particularly preferred.

The elastase inhibitor of the present invention can be used for various symptoms and diseases caused by denaturation and overproduction of elastin, for example, improvement of skin elasticity, suppression of wasted hair, improvement of inflammation,
Treatment of lupus and psoriasis, treatment of rheumatoid arthritis, improvement of respiratory diseases (emphysema, pulmonary fibrosis, pneumonia, bronchitis, bronchiectasis, asthma, etc.), cardiovascular diseases (arteriosclerosis, arteritis, myocardial infarction) Etc.), renal failure / liver failure, periodontitis, organ transplant rejection, etc. can be administered as a skin external preparation, oral preparation, injection, etc.

The elastase inhibitor of the present invention may contain, in addition to the above-mentioned active ingredients, any components as long as the effects of the present invention are not impaired. Depending on the dosage form of the elastase inhibitory activity, any of the above-mentioned optional components are commonly blended in known skin external preparations, oral preparations, injections, etc., for example,
Purified water, alcohol, chelating agents, various oils, surfactants, emulsifiers, thickeners, preservatives, antioxidants, solvents, medicinal ingredients, powders, pigments, fragrances, and the like can be blended. In addition, the elastase inhibitor of the present invention may optionally contain an existing elastase inhibitor, a keratinizing agent, an ultraviolet absorber, an ultraviolet protective agent, collagen, a humectant, an anti-inflammatory agent, an antioxidant, and the like. be able to.

The elastase inhibitor of the present invention can be formed into various forms by a conventional method, and the form is not particularly limited, but may be a lotion, an emulsion, a cream, an ointment,
It is preferably in the form of a stick, a solution with an organic solvent or purified water, a pack, a gel, or the like. That is, the external preparation for skin of the present invention comprises lotion, oil essence, O / W
Type or W / O type cream, emulsified skin external preparation, pack, ointment, foundation, skin cleanser, tonic,
Used as a bath agent and the like.

The elastase inhibitor of the present invention can be obtained by mixing and stirring the compound represented by the general formula (1) and other additives by a conventional method.

[0027]

EXAMPLES Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited to the following examples.

Synthesis Example 1 Ni-butoxycarbonyl-
Synthesis of L-leucinamide (Compound 1) 5.00 g (30 mmol) of L-leucinamide hydrochloride and 5.46 g (54 mmol) of triethylamine are dissolved in a mixed solvent of 150 ml of tetrahydrofuran and 50 ml of distilled water, and cooled to 5 ° C. on ice. Cool. At the same temperature, 3.29 g (33 mmol) of isobutyl chloroformate was added dropwise. After completion of the addition, the disappearance of the raw materials was confirmed by thin-layer chromatography, and 6.53 g (57 mmol) of 85% phosphoric acid was added to terminate the reaction. did. After distilling off tetrahydrofuran from the reaction mixture under reduced pressure, 300 ml of ethyl acetate was added for extraction, and the organic layer was washed with distilled water and saturated saline, and dried over anhydrous sodium sulfate. Then, the solvent was distilled off, and the residue was subjected to column chromatography, and extracted with a mixed solvent of ethyl acetate-n-hexane. The solvent was distilled off and Ni-butoxycarbonyl-L-leucinamide (Compound 1) 4.
82 g (80% yield) were obtained. The physical properties of the obtained compound are as follows.

NMR (CDCl ₃ ) δ: 0.85 (d, 3H, J = 5 Hz), 0.88 (d,
3H, J = 5Hz), 1.33-1.93 (m, 4H), 3.73 (d, 2H, J = 7Hz), 3.84-
4.00 (m, 1H), 6.93 (s, 1H), 7.07 (d, 1H, J = 9Hz), 7.26 (s, 1
H)

Synthesis Example 2 Synthesis of N-carboxy-L-phenylalanine amide (compound 2) 6.00 g (30 mm) of L-phenylalanine amide hydrochloride
ol) and 5.46 g (54 mmol) of triethylamine were dissolved in a mixed solvent of 150 ml of tetrahydrofuran and 50 ml of distilled water, and cooled to 5 ° C. with ice. At the same temperature, 3.29 g (33 mmol) of ethyl chloroformate was added dropwise. After completion of the addition, the disappearance of the raw materials was confirmed by thin-layer chromatography, and 6.53 g (57 mmol) of 85% phosphoric acid was added to terminate the reaction. did. After distilling off tetrahydrofuran from the reaction mixture under reduced pressure, 300 ml of ethyl acetate was added for extraction.
The organic layer was washed with distilled water and saturated saline, and dried by adding anhydrous sodium sulfate. The solvent was distilled off, the residue was dissolved in ethanol (100 ml), and a solution obtained by dissolving 1.35 g (24 mmol) of potassium hydroxide in distilled water (100 ml) was added with good stirring, followed by stirring at 70 ° C. for 3 hours to complete the reaction. .
The solvent was distilled off, and the residue was recrystallized from ethanol-diethyl ether to give 5.91 g of N-carboxy-L-phenylalanine amide (compound 2) as pale yellow crystals (yield: 8).
1%). The physical properties of the obtained compound are as follows.

NMR (DMSO-d ₆ ) δ: 2.58 (dd, 1H, J = 8.14 Hz),
2.95 (dd, 1H, J = 4.14Hz), 3.45 (br.s, 2H), 3.70-3.75 (m, 1
H), 6.07 (br.s, 1H), 7.13-7.28 (m, 5H)

Synthesis Example 3 Synthesis of N-dibenzyloxyphosphonyl-L-leucinamide (Compound 3) 6.0 g (36 mmol) of L-leucinamide hydrochloride and 14.6 g (144 mmol) of triethylamine were dissolved in 200 ml of chloroform. The mixture was cooled to 0 ° C. with good stirring. Further, 16.0 g of dibenzyl chlorophosphate (54 mm
ol) and the mixture was stirred at the same temperature for 20 hours to complete the reaction. Then, 50 ml of ethanol was added, and the mixture was stirred for 30 minutes. The reaction mixture was washed with 5% hydrochloric acid, distilled water, 10% aqueous sodium hydrogen carbonate and saturated saline in this order, and dried over anhydrous sodium sulfate. Thereafter, the solvent was distilled off under reduced pressure, the residue was subjected to column chromatography, and chloroform-
The mixture was extracted with a mixed solvent of methanol. The solvent was distilled off, and the residue was recrystallized from ethyl acetate-n-hexane to obtain 9.3 g (yield: 66%) of N-dibenzyloxyphosphonyl-L-leucinamide (compound 3) as white crystals. . The physical properties of the obtained compound are as follows.

NMR (CDCl ₃ ) δ: 0.84 (d, 3H, J = 4 Hz), 0.87 (d,
3H, J = 4Hz), 1.41-1.82 (m, 3H), 3.70-3.97 (m, 2H), 4.98
(d, 2H, J = 4Hz), 5.02 (d, 2H, J = 4Hz), 6.93 (s, 1H), 7.07 (d,
1H, J = 9Hz), 7.26 (s, 1H)

Synthesis Example 4 Synthesis of N-phosphono-L-leucinamide dipotassium salt (Compound 4) 5.0 g (13 mmol) of N-dibenzyloxyphosphonyl-L-leucinamide (Compound 3) was added to methanol 100
dissolved in 3.3 ml of 8N aqueous potassium hydroxide solution.
% Palladium carbon was added, and the mixture was stirred for 2 hours under a hydrogen stream at room temperature to complete the reaction. After the reaction mixture was filtered to remove the catalyst, the solvent was distilled off, and the residue was diluted with ethanol.
Recrystallization from diethyl ether gave 3.2 g of N-phosphono-L-leucinamide dipotassium salt (compound 4) (yield 97).
%). The physical properties of the obtained compound are as follows.

NMR (D ₂ O) δ: 0.94 (d, 6H, J = 6 Hz), 1.47-1.61
(m, 2H), 1.69-1.79 (m, 1H), 3.43-3.76 (m, 1H) and amide proton were not observed.

The following compounds were obtained in the same manner as in Synthesis Examples 1 to 4.

[0037]

[Table 1]

Test Example 1 Test for Elastase Activity Inhibition of Cultured Human Fibroblasts Normal human fibroblasts commercially available from Dainippon Pharmaceutical Co., Ltd. were subcultured in a DME medium containing 10% fetal bovine serum, and used in this test. did. The cells detached from the petri dish using a rubber policeman were suspended in saline, the cells were collected using a low-speed centrifuge, and washed three times with saline. Cells are 0.1% Triton X-100 / 0.2M Tris-HCl buffer
The suspension was suspended in [pH 8.0] and sonicated to prepare an enzyme solution.
As a substrate for measuring the enzyme activity, 125 mM N-Suc- (Ala) 3-p-nitroanilide was used. Compounds 1 to 14 were respectively added to the enzyme solution, and 1 μl of the substrate was added to 100 μl of the enzyme solution + test subject adjusted to have the evaluation concentrations shown in Table 2.
For 1 hour, and the reaction was stopped by adding 5 μl of acetic acid. The amount of nitroaniline formed was 405 on a spectrophotometer.
The absorbance at nm was measured to determine the elastase activity inhibition rate. Table 2 shows the results.

[0039]

[Table 2]

From Table 2, it was found that Compounds 1 to 14 had a remarkable elastase activity inhibitory action.

[0041]

The elastase inhibitor of the present invention has an excellent elastase activity inhibitory activity and the like, and is easily synthesized.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 31/195 ADA A61K 31/195 ADA 31/22 31/22 31/40 31/40 31/66 31/66 // C07D 207 / 16 C07D 207/16 (72) Inventor Yoshinori Nishizawa 2606 Akabane, Kakaicho, Haga-gun, Tochigi Pref.

Claims (2)

[Claims] In the following general formula (1),(Where X is COOR^Two(Where R^TwoIs a hydrogen atom or
Shows an alkyl group. ), The following general formula (2):(Where R^ThreeAnd R^FourRepresents a hydrogen atom, an alkyl group or
Represents a alkyl group. ), The following general formula (3):(Where R^FiveIs a hydrogen atom, alkyl group, or aralkyl
A group represented by R⁶Represents an aralkyl group. ) Or SO_ThreeH
And Y is N (R¹') (Where R¹'Is a hydrogen atom or
Shows a lower alkyl group. ), O or CH_TwoAnd Z is
CON (R⁷) R⁸ Or COOR⁹(Where R⁷, R⁸Passing
And R⁹Represents a hydrogen atom or an alkyl group. ) And R
¹Is a hydrogen atom, an alkyl group which may have a substituent,
An aralkyl group or an alkoxyl group, or R¹When
R¹′ Together form a ring. )
Elaster containing the compound described above or a salt thereof as an active ingredient
Ze inhibitor. 2. In the general formula (1), X is COOR
² (wherein, R ² is the same as described above);
Is N (R ¹ ′) (wherein R ¹ ′ is the same as described above), Z is CONH ₂ , and R ¹ has 3 carbon atoms.
5 to 5 are alkyl, benzyl, carbamoylmethyl, 2-carbamoylethyl and hydroxymethyl, wherein R ¹ ′ is a hydrogen atom or R ¹ and R ¹ ′ together form a ring. An elastase inhibitor according to claim 1.