JP2008019188A - New carnosine derivative and composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new L-carnosine derivative which overcomes the difficult points of L-carnosine, is stable to carnosinase, and is excellent in an anti-oxidizing action. <P>SOLUTION: This L-carnosine derivative is represented by the general formula (II), or its pharmacologically acceptable salt, ester or amide. This compound can be used for medicines, skin cosmetics and the like for treating or preventing diseases caused by damages due to active oxygen species, for preventing ageing, and the like. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a novel L-carnosine derivative which is stable against carnosinase and excellent in antioxidant activity, and a composition containing the derivative.

  Peroxidation of lipids and proteins caused by reactive oxygen species is considered to be one of the factors that decrease the structure and function of human and other mammalian tissues with aging. Substances to prevent such peroxidation As an antioxidant, it is applied to pharmaceuticals, cosmetics, dietary supplements, and so on.

で示されるＬ−カルノシン（β−アラニル−Ｌ−ヒスチジン）が知られている（例えば、特許文献１）。 Examples of such substances are those of formula (I):

L-carnosine (β-alanyl-L-histidine) represented by formula (1) is known (for example, Patent Document 1).

  L-carnosine is a natural antioxidant abundant in non-protein fractions of vertebrate skeletal muscle (for example, Non-patent Documents 1 and 2), lens (for example, Non-patent Document 3), and the like. Middle component lipid, protein, etc. are acrolein, secondary produced by oxidation of reactive oxygen species, lipid peroxide (for example, Non-patent Documents 4 and 5) or unsaturated fatty acid, 4-hydroxy-trans-2,3 -It is known to play an important role in preventing damage by α, β-unsaturated aldehydes such as nonenal (HNE) (eg, Non-Patent Documents 6 and 7). Therefore, L-carnosine is used as a dietary supplement and cosmetics in some countries. However, it is easily hydrolyzed by an enzyme called carnosinase, which is present in a large amount in plasma or tissue, and β-alanine and L- Since it becomes histidine and loses its physiological activity completely, it is not used as a pharmaceutical product, and products such as nutritional supplements and cosmetics mainly composed of L-carnosine are not necessarily satisfactory. In addition, as far as the present inventors know, there is no derivative that is chemically modified from L-carnosine and is used for the above purpose.

JP 2003-267992 A K. G. Crush, Com. Biocem. Physiol., Vol. 34, 3, 1970 A. A. Boldyrev., S. E. Severin, Adv. Enzyme Regul., Vol. 30, 1990 F. Margolis, Science, Vol.184, 909, 1974 M. A. Babizhayev et al., Biochem. J., Vol. 304, 509, 1994 J. H. Kang et al., Molecules and Cells, Vol. 13, 107, 2002 G. Poli, R. Schaur, IUBMB Life, Vol.50, 315, 2000 F. J. Romero et al., Environ. Health Perspect., Vol.106, 1229, 1998

  An object of the present invention is to provide a novel L-carnosine derivative that overcomes the difficulties of L-carnosine, is stable to carnosinase, and has an excellent antioxidant action. Another object of the present invention is to provide a composition that can be applied to this novel L-carnosine derivative for pharmaceuticals, skin cosmetics, etc. for the purpose of treating or preventing diseases caused by disorders caused by reactive oxygen species and preventing aging. It is to provide.

  The present inventors pay attention to L-carnosine (that is, β-alanyl-L-histidine), which is a natural antioxidant present in a large amount in various animal living bodies and is excellent in safety, and has an antioxidant activity. As a result of intensive research aimed at developing a novel L-carnosine derivative that is superior to L-carnosine and stable against peptidases such as carnosinase, the inventors succeeded in creating a novel carnosine derivative. In addition, the present inventors have found that the carnosine derivative exhibits stronger antioxidant activity than L-carnosine and is superior in resistance to proteases such as carnosinase, and the derivative has UV protection superior to L-carnosine. The present invention was completed by finding that it has an action and a moisturizing action on the skin, and further studied.

で示されるＬ−カルノシン誘導体、またはその薬理的に許容し得る塩、エステルもしくはアミド、
［２］ Ｌ−カルノシン誘導体（ＩＩ）が、Ｎ−［（２ＲＳ）−６−ヒドロキシ−２，５，７，８−テトラメチルクロマン−２−カルボニル］−Ｌ−カルノシンである前記［１］記載の化合物、
［３］ Ｌ−カルノシン誘導体（ＩＩ）が、Ｎ−［（２Ｒ）−６−ヒドロキシ−２，５，７，８−テトラメチルクロマン−２−カルボニル］−Ｌ−カルノシンである前記［１］記載の化合物、
［４］ Ｌ−カルノシン誘導体（ＩＩ）が、Ｎ−［（２Ｓ）−６−ヒドロキシ−２，５，７，８−テトラメチルクロマン−２−カルボニル］−Ｌ−カルノシンである前記［１］記載の化合物、
［５］ 前記［１］〜［４］のいずれかに記載の化合物を含有する医薬、
［６］ 前記［１］〜［４］のいずれかに記載の化合物を含有する化粧品、および
［７］ 前記［１］〜［４］のいずれかに記載の化合物を含有する抗酸化剤、
に関する。 That is, the present invention
[1] Formula (II):

Or a pharmacologically acceptable salt, ester or amide thereof,
[2] The above-mentioned [1], wherein the L-carnosine derivative (II) is N-[(2RS) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine. A compound of
[3] The above [1], wherein the L-carnosine derivative (II) is N-[(2R) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine. A compound of
[4] The above [1], wherein the L-carnosine derivative (II) is N-[(2S) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine. A compound of
[5] A medicament containing the compound according to any one of [1] to [4],
[6] A cosmetic containing the compound according to any one of [1] to [4], and [7] an antioxidant containing the compound according to any one of [1] to [4],
About.

  According to the present invention, the novel L-carnosine derivative (II) has a stronger antioxidant activity than natural L-carnosine and resists hydrolysis by proteases such as carnosinase, so that it can be administered orally or parenterally. It can be stably present in various tissues of the living body, and is useful for the prevention and treatment of disorders caused by reactive oxygen species such as human or other mammalian tissues and skin. Therefore, it can be used for the prevention and treatment of ischemic diseases such as arteriosclerosis, myocardial infarction, cerebral infarction, subarachnoid hemorrhage, and can be expected to have an anti-aging effect, as well as UV protection and water retention It is also useful as a cosmetic product that promotes skin health.

で示されるＬ−カルノシン誘導体、またはその薬理的に許容し得る塩、エステルもしくはアミドである。 The target compound of the present invention has the general formula (II):

Or a pharmacologically acceptable salt, ester or amide thereof.

  Examples of the “pharmacologically acceptable salt” include alkali metal salts such as sodium salt and potassium salt, inorganic acid salts such as hydrochloride and sulfate, malate, fumarate and succinate. And organic acid salts such as tartrate.

  The “ester” includes an ester that is a “general protecting group” of a carboxyl group, or an ester that is a “protecting group that can be cleaved in vivo by a biological method such as hydrolysis”.

  Examples of the “general protecting group” include an alkyl group such as a methyl group, an ethyl group, a propyl group and a butyl group; an aralkyl group such as a benzyl group and a phenethyl group; a hydroxyethyl group and a 3-hydroxypropyl group. And such hydroxy-substituted alkyl groups.

  Examples of the “protecting group that can be cleaved by biological methods such as hydrolysis in vivo” include, for example, methoxymethyl group, 1-ethoxyethyl group, 1-methyl-1-methoxyethyl group, ethoxymethyl group , N-propoxymethyl group, alkoxy-substituted alkyl group such as tert-butoxymethyl group; alkoxycarbonyl-substituted alkyl group such as methoxycarbonylmethyl group; alkylthio-substituted alkyl group such as methylthiomethyl group and ethylthiomethyl group; acetoxy Methyl, 1-acetoxyethyl, propionyloxymethyl, 1-propionyloxyethyl, butyryloxymethyl, pivaloyloxymethyl, 1-pivaloyloxyethyl, valeryloxymethyl, iso Valeryloxymethyl group, 1-cyclohexanecarbo Aliphatic acyloxy substituted alkyl groups such as Ruokishiechiru group; methoxycarbonyloxymethyl group, ethoxycarbonyloxymethyl group, an alkoxycarbonyl-substituted alkyl groups such as propoxycarbonyl oxymethyl group.

［式中、ＲａおよびＲｂはそれぞれ独立して水素原子、アルキル基またはアラルキル基を示すか、あるいは両者が隣接する窒素原子と共に５〜７員複素環式基を形成していてもよいことを示す。］
で示される基が挙げられる。 Examples of the group that forms “the amide” include, for example, the formula (A):

[Wherein, Ra and Rb each independently represent a hydrogen atom, an alkyl group or an aralkyl group, or they may form a 5- to 7-membered heterocyclic group together with an adjacent nitrogen atom. . ]
The group shown by these is mentioned.

［式中、Ｒは水素原子またはカルボキシル基の保護基を示す。］
で示されるＬ−カルノシン類化合物と一般式（ＩＩＩ）：

［式中、Ｒ’は水素原子または水酸基の保護基を示す。]
で示される６−ヒドロキシ−２，５，７，８−テトラメチルクロマン誘導体またはそのカルボキシル基における反応性誘導体を反応させ、Ｒおよび／またはＲ’が保護基である場合は、さらに生成物から該保護基を除去するか、あるいは
（方法２）
一般式（ＩＶ）：

［式中、Ｒは上記と同意義を示す。］
で示されるＬ−ヒスチジン類化合物と式（Ｖ）：

［式中、Ｒ’は上記と同意義を示す。］
で示される化合物またはそのカルボキシル基における反応性誘導体を反応させ、Ｒおよび／またはＲ’が保護基である場合は、さらに生成物から該保護基を除去することにより、目的化合物（ＩＩ）を製造することができる。 The novel L-carnosine derivative (II) according to the present invention can be produced, for example, by the following method 1 or method 2.
(Method 1)
Formula (Ia):

[Wherein, R represents a hydrogen atom or a protecting group for a carboxyl group. ]
L-carnosine compounds represented by general formula (III):

[Wherein, R ′ represents a hydrogen atom or a protecting group for a hydroxyl group. ]
When the 6-hydroxy-2,5,7,8-tetramethylchroman derivative represented by or a reactive derivative at its carboxyl group is reacted and R and / or R ′ is a protecting group, Removing the protecting group or (Method 2)
Formula (IV):

[Wherein, R represents the same meaning as described above. ]
L-histidine compounds represented by the formula (V):

[Wherein, R ′ is as defined above. ]
In the case where R and / or R ′ is a protecting group, the target compound (II) is produced by further removing the protecting group from the product. can do.

［式中、Ｒは前記と同意義を示す。］
で示されるβ−アラニン類化合物と上記化合物（ＩＩＩ）またはそのカルボキシル基における反応性誘導体を反応させ、次いで必要により脱保護することにより化合物（Ｖ）を製造することができる。 The starting compound (V) in Method 2 can be produced as follows. That is, the formula (VI)

[Wherein, R represents the same meaning as described above. ]
The compound (V) can be produced by reacting the β-alanine compound represented by formula (II) with the compound (III) or a reactive derivative thereof at the carboxyl group and then deprotecting as necessary.

  In the compounds represented by the formulas (Ia), (IV) and (VI), when the group represented by R is a protecting group for a carboxyl group, the protecting group may be a carboxylic acid protecting group in the field of amino acid or peptide chemistry. A well-known thing is mentioned as group. Examples of such protecting groups include alkyl groups (eg, methyl group, ethyl group, propyl group, butyl group, tert-butyl group) and aralkyl groups (eg, benzyl group, p-methoxybenzyl group, etc.). It is done.

  On the other hand, in the compounds represented by the formulas (III) and (V), examples of the protecting group for the hydroxyl group represented by R ′ include an acyl group (for example, a formyl group, an acetyl group, a benzoyl group) and an alkoxycarbonyl group (for example, Methoxycarbonyl group, ethoxycarbonyl group, propoxycarbonyl group, butoxycarbonyl group, isobutoxycarbonyl group, tert-butoxycarbonyl group, etc.), aralkyl groups (for example, benzyl group, p-methoxybenzyl group, etc.) and the like.

Reaction of Compound (III) with Compound (Ia) or Reaction of Compound (V) with Compound (IV) :
These reactions are usually carried out in the presence of an appropriate condensing agent, or once compound (III) or compound (V) is once led to a reactive derivative at its carboxyl group, and then reacted with compound (Ia) or compound (IV), respectively. Is done. As the condensing agent used in the above reaction, those known per se, such as dicyclohexylcarbodiimide (DCC), water-soluble carbodiimide (WSC) [for example, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride, etc.] , Carbonyldiimidazole (CDI), benzotriazol-1-yloxy-tris (pyrrolidino) phosphonium hexafluorophosphate (PyBOP), diphenylphosphoric acid azide (DPPA), etc., which are optionally N-hydroxysuccinimide ( HOSu), 1-hydroxybenzotriazole (HOBT) and the like may be used together. Examples of the reactive derivative in the carboxyl group of compound (III) or compound (V) include acid chloride, active ester (for example, p-nitrophenyl ester, pentachlorophenyl ester, ester with N-hydroxysuccinimide, Esters with 1-hydroxybenzotriazole, etc.), imidazolides, mixed acid anhydrides (eg methoxy formic acid, ethoxy formic acid, propoxy formic acid, butoxy formic acid, isobutoxy formic acid, tert-butoxy formic acid, phenoxy formic acid, 2,2-dimethylpropionic acid Methanesulfonic acid, benzenesulfonic acid, mixed acid anhydrides with toluenesulfonic acid, etc.). These reactive derivatives may be once isolated and reacted with the compound (Ia) or the compound (IV). After the reactive derivative is produced in the reaction system or while being produced, the compound (Ia) or (IV ).

  The above reaction is usually performed in a solvent in the presence of a base as necessary. As the solvent, an inert organic solvent is usually used, but in some cases, water can be used as a solvent, or a mixture thereof can also be used. Examples of the organic solvent used include alkyl halides (eg, methylene chloride, chloroform, etc.), aromatic hydrocarbons (eg, benzene, toluene, xylene, etc.), ethers (eg, diethyl ether, diisopropyl ether, methyl isobutyl). Ether, methylcyclopentyl ether, tetrahydrofuran, dioxane, etc.), esters (eg, methyl acetate, ethyl acetate, isopropyl acetate, butyl acetate, etc.), ketones (eg, acetone, methyl ethyl ketone, methyl isobutyl ketone, etc.), N, N- Examples include dimethylformamide, N, N-dimethylacetamide, N-methylpiperidone, dimethyl sulfoxide and the like. Examples of the base that can be used include inorganic bases (for example, sodium hydrogen carbonate, potassium hydrogen carbonate, sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, lithium hydroxide, etc.), organic bases (for example, pyridine, triethylamine, N, N-diisopropylethylamine, N-methylmorpholine, N-methylpiperidine, etc.). The reaction temperature varies depending on the condensing agent used, the reactive derivative of compound (III) or the reactive derivative of compound (V), etc., but is usually about −30 ° C. to 100 ° C., preferably −10 ° C. to about It is in the range of 40 ° C. The amount of the condensing agent and base used is usually 1 to 5 equivalents, preferably 1 to 3 equivalents, per 1 mol of compound (III) or compound (V). When compound (III) or (V) is used in the form of a reactive derivative thereof, the amount used is 0.1 to 3 equivalents, preferably 0.1 to 1 mol of compound (IV) or compound (VI). 3 to 1 equivalent.

  When R and / or R ′ in the compound thus obtained is a protective group other than a hydrogen atom, appropriate reaction conditions known per se (eg, hydrolysis with an acid or alkali) depending on the protective group. The target compound (II) can be obtained by deprotection using decomposition, catalytic reduction using palladium carbon as a catalyst, or the like. The target compound (II) can be isolated and purified by, for example, solvent extraction, chromatography, recrystallization and the like.

  When the raw material compound (V) is produced from the compound (III) and the compound (IV), it can be carried out in the same manner as in the above condensation reaction and deprotection reaction.

  In addition, since the starting compound (III) or (V) has an asymmetric carbon, there are two kinds of optical isomers [(R) isomer and (S) isomer]. Therefore, depending on the optical isomers of the compound (III) or compound (V) used, the target compound (II) has two isomers each having a diastereomeric relationship, [(R)-(S)]. Or a racemic form of compound (III) or (V), the desired product (II) can be obtained as a mixture of the diastereomers, [(RS )-(S)] body. In addition, the diastereomeric mixture of the target product (II) can be separated and purified as each diastereomer if necessary. The object (II) of the present invention includes any of these diastereomers and mixtures thereof, but is optically pure [(R)-(S)] or [(S)- (S)] body is preferred.

  The compound (II) obtained as described above can be converted into a pharmacologically acceptable salt, ester or amide, if necessary. The salt formation reaction, esterification reaction and amidation reaction can be carried out according to conventional methods employed in this technical field. The ester can also be obtained as a synthetic intermediate in Method 1 or Method 2.

  The object compound (II) of the present invention obtained as described above or a pharmacologically acceptable salt, ester or amide thereof has excellent antioxidant activity and resists degradation by carnosinase or other proteases. Therefore, when administered orally or parenterally, it is hardly degraded in the human body or other mammals and acts as a stable antioxidant. Therefore, it can be used for the prevention and treatment of various diseases caused by reactive oxygen species such as arteriosclerosis, myocardial infarction, angina pectoris, cerebral infarction, subarachnoid hemorrhage, etc. Anti-aging effects against various aging phenomena can be expected. In addition, since the target compound (II) prevents or delays skin aging and has an ultraviolet protection effect, it can be blended into cosmetics and used as skin cosmetics.

  When the target compound of the present invention is used as a pharmaceutical, the pharmaceutical dosage form is, for example, a preparation for oral administration such as tablets, capsules, granules, powders and syrups, or parenteral administration such as injections and suppositories. Preparations. These preparations are made of excipients (eg lactose, sucrose, glucose, mannitol, sorbitol, corn starch, potato starch, crystalline cellulose, gum arabic, dextran, pullulan, light anhydrous silicic acid, synthetic aluminum silicate, calcium carbonate, phosphoric acid. Calcium hydrogen), lubricant (eg stearic acid, calcium stearate, magnesium stearate, talc, colloidal silica etc.), binder (eg hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, macrogol etc.), disintegration Agents (for example, low-substituted hydroxypropylcellulose, carboxymethylcellulose, carboxymethylcellulose calcium, carboxymethyl starch, carboxymethyl starch natri) And additives such as stabilizers (eg methyl paraben, propyl paraben, benzyl alcohol), flavoring agents (eg, commonly used sweeteners, acidulants, flavors), diluents, etc. And manufactured by a well-known method. The dose of the target compound varies depending on the type of disease, symptoms, age, administration method, etc. For example, in the case of oral administration, it is 0.1-100 mg / kg / day, particularly 0.5-20 mg / kg / day. Day is preferred. In the case of intravenous injection, 0.01 to 10 mg / kg / day, particularly 0.1 to 2 mg / kg / day is preferable.

  In addition, when the target compound of the present invention is used as a cosmetic, the cosmetic dosage form may be any ointment, cream, emulsion, lotion, oil, gel, etc., as long as it is suitable for application to the skin. The ointment form is preferred. The ointment can be prepared according to a known method using a known ointment base. Examples of the base include white petrolatum, yellow petrolatum, water-absorbing petrolatum, macrogol, paraffin, liquid paraffin, plastibase, silicone, beef tallow, wax, lanolin, vegetable oil, hydroxypropylcellulose, and the like. The formulations can contain conventional cosmetic additives, such as fragrances. Although the compounding quantity of the target compound in cosmetics is not specifically limited, Usually, 0.01-5 w / w%, especially 0.1-2 w / w% are preferable.

  Hereinafter, the present invention will be described in more detail with reference to Examples and Examples, but this does not limit the present invention. In the present specification, the “alkyl group” is preferably an alkyl group having 1 to 6 carbon atoms, the alkoxy group is preferably an alkoxy group having 1 to 6 carbon atoms, and the aliphatic acyl group has 1 carbon atom. ~ 6 aliphatic acyl groups are preferred. The chroman skeleton contained in the object compound (II) of the present invention is also named “3,4-dihydro-2H-benzopyran”.

(Reference Example 1) L-carnosine benzyl ester · 2 (p-toluenesulfonic acid) salt L-carnosine (raw compound (I)) (22.6 g, 0.1 mol) and p-toluenesulfonic acid monohydrate (Hereinafter abbreviated as TsOH · H ₂ O) (39.9 g, 0.21 mol) was dissolved in water (100 ml), and then water was distilled off under reduced pressure. To the residue, TsOH.H ₂ O (5.0 g, 0.026 mol) and benzyl alcohol (75 ml) were added and dissolved, and then chloroform (80 ml) was further added, and the resulting water was subjected to azeotropic conditions. While being removed with Molecular Sieves-4A, the mixture was heated to reflux for 12 hours until the spot of the raw material compound (I) disappeared by thin layer chromatography (hereinafter abbreviated as TLC).
Chloroform was distilled off under reduced pressure, and ethyl acetate (hereinafter abbreviated as AcOEt) (300 ml) was added to the residue to wash the residual oily substance. Further, this oily substance was heated to about 30 ° C., washed with AcOEt (4 × 300 ml), and then dried under reduced pressure to obtain the title compound (52.5 g, 79.7%) as an amorphous substance.
¹ H-NMR (200 MHz, DMSO-d ₆ ): δ 2.30 (6H, s), 2.45-2.56 (2H, m), 2.88-3.23 (4H, m), 4 60-4.74 (1H, m), 5.12 (2H, s), 7.08-7.16 (4H, brd), 7.28-7.53 (10H, m), 7. 68 (3H, brs), 8.99 (1H, d, J = 1.2 Hz).

Reference Example 2 N-tert-butoxycarbonyl-L-carnosine benzyl ester N-tert-butoxycarbonyl-β-alanine (1.5 g, 7.92 mmol) and L-histidine benzyl ester · 2 (p-toluenesulfonic acid) ) Salt (7.02 g, 11.9 mmol) in N, N-dimethylformamide (hereinafter abbreviated as DMF) (20 ml) in ice-cooled N, N-diisopropylethylamine (hereinafter abbreviated as DIEA) (3. 07 g, 23.8 mmol), followed by 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1.67 g, 8.72 mmol), and then stirred at room temperature for 72 hours. The reaction was diluted with AcOEt (40 ml) and washed with water (1 × 40 ml, 2 × 10 ml) and saturated brine (20 ml). The aqueous layer was further extracted with AcOEt (2 × 15 ml), then the entire organic layer was dried (MgSO ₄ ) and the solvent was distilled off. The oily residue was separated and purified by silica gel column chromatography using hexane / ethyl acetate (1: 1) and then chloroform / methanol (15: 1) as a solvent. The title compound (3.14 g, 95%) was obtained. Obtained as a colorless amorphous material.
¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.43 (9H, s), 2.42 (2H, br dd), 3.06 (1H, do of a pair of ABq, J _AB = 15.6 Hz) , J _AX = 4.9 Hz), 3.12 (1H, d of a pair of ABq, J _AB = 15.6 Hz, J _BX = 4.9 Hz), 3.34-3.49 (2H, m), 4.84 (1H, dt, J = 7.5, 4.9 Hz), 5.08 and 5.17 (1Heach, ABq, J = 12.2 Hz), 5.59 (2H, br t), 6 .53 (1H, d, J = 1.1 Hz), 7.22 (1H, br d, J = 7.5 Hz), 7.25-7.40 (5H, m), 7.50 (1H, d , J = 1.1 Hz).

(Reference Example 3) A solution of L-carnosine benzyl ester dihydrochloride N-tert-butoxycarbonyl-L-carnosine benzyl ester (1.00 g, 2.40 mmol) in acetonitrile (5 ml) in 4N hydrogen chloride-AcOEt solution (3 ml) ) Was added dropwise at room temperature, followed by stirring at the same temperature for 10 minutes. The reaction solution was concentrated under reduced pressure to obtain L-carnosine benzyl ester dihydrochloride (940 mg, quantitative).
¹ H-NMR (200 MHz, CD ₃ OD): δ 2.69 (2H, t, J = 6.4 Hz), 3.10-3.38 (4H, m), 4.73-4.94 (1H , M), 5.15 and 5.22 (1H each, ABq, J = 12.1 Hz), 7.27 (1H, d, J = 1.3 Hz), 7.30-7.44 (5H, m ), 8.78 (1H, d, J = 1.3 Hz).

Reference Example 4 (±) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl-β-alanine (compound (V), R ′ = H)
(±) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (compound (III), R ′ = H) (1.00 g, 3.99 mmol) and β-alanine ethyl ester A solution of hydrochloride (736 mg, 4.79 mmol) in DMF (10 ml) was added DIEA (2.07 g, 16.0 mmol), followed by benzotriazol-1-yloxy-tris (pyrrolidino) phosphonium hexafluorophosphate (hereinafter PyBOP). Abbreviated) (2.08 g, 3.99 mmol). Thereafter, the reaction solution was stirred overnight at room temperature. It was confirmed by TLC that the raw material (compound (III), R ′ = H) had disappeared. The reaction mixture was diluted with AcOEt (30 ml), washed with 1N HCl (2 × 20 ml), saturated aqueous sodium hydrogen carbonate solution (1 × 15 ml) and saturated brine (20 ml), and dried (MgSO ₄ ). After distilling off the solvent, the crude product was purified by silica gel column chromatography using hexane / ethyl acetate (3: 1) as a solvent. The ethyl ester of the title compound (1.42 g, quantitative) was obtained as a yellowish white amorphous substance. Obtained as material.
¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.21 (3H, t, J = 7.1 Hz), 1.49 (3H, s), 1.88 (1H, ddd, J = 13.5, 8.1, 6.6 Hz), 2.09 (3H, s), 2.18 (6H, s), 2.24-2.72 (5 H, m), 3.49 (2H, q, J = 6.1 Hz), 4.05 and 4.06 (total 2H, both q, J = 7.1 Hz), 4.50 (1H, s), 7.05 (1H, br t, J = 6.1 Hz) .

A solution of lithium hydroxide monohydrate (0.50 g, 12.0 mmol) in water (4 ml) was added dropwise to a solution of the ethyl ester (1.40 g, 3.99 mmol) in methanol (10 ml) and refluxed for 2 hours. . After the methanol was distilled off under reduced pressure, the liquidity of the residual liquid was adjusted to pH 1 with 1N hydrochloric acid. The precipitated crystals were extracted with AcOEt (1 × 50 ml, 2 × 10 ml), the organic layer was dried (MgSO ₄ ), and the solvent was distilled off to obtain the title compound (1.24 g, 96.5%) as a white amorphous substance. It was.
¹ H-NMR (200 MHz, CD ₃ OD): δ 1.45 (3H, s), 1.82 (1H, ddd, J = 13.4, 8.6, 6.4 Hz), 2.06 (3H , S), 2.14 (6H, s), 2.21-2.71 (5H, m), 3.34-3.47 (2H, m), 7.50 (1H, br t, J = 5.9 Hz).

(Example 1)
1) N-[(2RS) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine benzyl ester (R: S = 1: 1 mixture)
Method A : (±) -6-hydroxy-2,5,7 in a solution of L-carnosine benzyl ester · 2 (p-toluenesulfonic acid) salt (0.86 g, 1.3 mmol) in pyridine (5 ml) at room temperature , 8-tetramethylchroman-2-carboxylic acid (250 mg, 1.0 mmol) was added, followed by 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (280 mg, 1.46 mmol) at 5-10 ° C. ) And the reaction was stirred at room temperature for 6 hours. After pyridine was distilled off under reduced pressure, the residue was dissolved in AcOEt (20 ml), washed with saturated brine (2 × 20 ml), and dried (MgSO ₄ ). After the solvent was distilled off under reduced pressure, the residue was purified by silica gel column chromatography using hexane / ethyl acetate (1: 1) and then chloroform / methanol (10: 1) as a solvent to give the title compound (260 mg, 56 .8%) was obtained as a viscous oil.
¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.48 and 1.51 (total 3H, both), 1.70-2.70 (6H, m), 2.07 and 2.08 (total 3H) , Both s), 2.14 and 2.17 (total 6H, both s), 2.80-3.10 (2H, m), 3.20-3.67 (2H, m), 4.38 ( 0.5H, dt, J = 6.8, 6.1 Hz), 4.73 (0.5H, dt, J = 7.5, 5.7 Hz), 5.03 and 5.09 (1Heach, ABq) , J = 12.3 Hz), 6.46 (0.5 H, br d, J = 6.8 Hz), 6.53 (0.5 H, d, J = 1.0 Hz), 6.57 (0.5 H) , D, J = 0.9 Hz), 7.10-7.40 (6.5 H, m), 7.45 (0.5 H, , J = 0.9Hz), 7.49 (0.5H, d, J = 1.0Hz).

Method B : (±) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl-β-alanine (compound (V), R ′ = H) (600 mg, 1.87 mmol) and L -A solution of histidine benzyl ester 2 (p-toluenesulfonic acid) (3.30 g, 5.60 mmol) in DMF (10 ml) at room temperature with DIEA (1.21 g, 9.34 mmol) followed by 1- Ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (394 mg, 2.05 mmol) was added, and the mixture was stirred at the same temperature for 96 hours. The reaction mixture was diluted with AcOEt (40 ml), washed with water (1 × 20 ml, 4 × 10 ml) and saturated brine (15 ml), dried (MgSO ₄ ), and the solvent was evaporated. The oily residue was separated and purified by silica gel column chromatography using hexane / ethyl acetate (1: 1) and then chloroform / methanol (10: 1) as a solvent to give the title compound (869 mg, 85%) as a viscous oil. Obtained.

2) N-[(2RS) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine compound (1)
N-[(2RS) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine benzyl ester (R: S = 1: 1 mixture) (895 mg, 1.63 mmol) 10% palladium carbon (water content: 52.7%) (200 mg) was added to a methanol (8 ml) solution, and catalytic reduction was performed at normal temperature and normal pressure. After removing the catalyst by filtration, the solvent was distilled off to obtain a slightly yellow viscous oily substance. This was dissolved in methanol / chloroform (1: 3), AcOEt was gradually added to crystallize the product, and then the supernatant was removed (this operation was repeated three times). After distilling off the solvent, the oily residue was dissolved in methanol (1.5 ml), and ligroin (boiling range 80-110 ° C., 4-5 ml) was gradually added to form a suspension. Thereafter, the solvent was distilled off under reduced pressure, and the residue was further dried under vacuum to obtain the title compound (617 mg, 82%) as a slightly yellowish white amorphous substance.
¹ H-NMR (200 MHz, CD ₃ OD): δ 1.436 and 1.444 (total 3H, both s), 1.75-1.90 (1H, m), 2.05 (3H, s), 2.13 (3H, s), 2.14 (3H, s), 2.10-2.70 (5H, m), 2.99-3.52 (4H, m), 4.64-4. 72 (1H, m), 7.29 (0.5 H, d, J = 1.3 Hz), 7.30 (0.5 H, d, J = 1.3 Hz), 7.59 (0.5 H, br t), 7.90 (0.5 H, s), 8.74 (0.5 H, d, J = 1.3 Hz), 8.81 (0.5 H, d, J = 1.3 Hz).
Electrospray ionization mass spectrometry Positive ion mode:
Observation (M + H) ⁺ : 459.1 for C ₂₃ H ₃₁ N ₄ O ₆
Calculated value: 459.2 for isotopic model MH;
Anion mode:
Observation (M-H) ⁻ : 457.1 for C ₂₃ H ₂₉ N ₄ O ₆
Calculated value: 457.2 for isotopic model (M-H).
As a result of HPLC analysis (below), the compound (1) was a mixture of two diastereomers (1: 1).

(Example 2)
1) N-[(2R) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine benzyl ester L-carnosine benzyl ester dihydrochloride (1.12 g, 2 .88 mmol) in DMF (3 ml) at room temperature with (2R) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (721 mg, 2.88 mmol) and dichloromethane (3 ml). After addition and dissolution, DIEA (1.12 g, 8.64 mmol) and PyBOP (1.50 g, 2.88 mmol) were added, and the reaction solution was stirred at room temperature for 72 hours. After most of the dichloromethane was distilled off under reduced pressure, the residue was dissolved in AcOEt (40 ml), and this was washed with water (1 × 30 ml, 3 × 10 ml) and saturated brine (20 ml) (aqueous layer pH: 5.5). The aqueous layer was further extracted with AcOEt (2 × 10 ml), then the entire organic layer was dried (MgSO ₄ ) and concentrated. The oily residue was separated and purified by silica gel column chromatography using chloroform and then chloroform / methanol (15: 1) as a solvent to give the title compound (1.58 g, quantitative) as a white amorphous substance.
¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.50 (3H, s), 1.69-1.88 (1H, m), 2.05-2.67 (5H, m), 2.07 (3H, s), 2.15 (6H, s), 2.90 and 2.97 (1 Heach, ABX system, J _AB = 15.6 Hz, J _AX = 6.1 Hz), 3.21-3. 38 (1H, m), 3.49-3.67 (1H, m), 4.46 (1H, dt, J = 6.8, 6.1 Hz), 5.04 and 5.10 (1H each, ABq, J = 12.3 Hz), 6.33 (2H, br), 6.49 (1H, d, J = 1.0 Hz), 6.54 (1H, d, J = 6.8 Hz), 7. 16 (1H, dd, J = 7.0, 5.2 Hz), 7.22-7.37 (5H, m), 7.49 (1H, d, J = 1) .0Hz).

2) N-[(2R) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine compound (2)
10% palladium in a solution of N-[(2R) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine benzyl ester (700 mg, 1.27 mmol) in methanol (3 ml). Carbon (water content: 52.7%) (100 mg) was added, and catalytic reduction was performed at normal temperature and normal pressure. Thereafter, the catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain a yellowish white amorphous. AcOEt (5 ml) was added to this and stirred, and then the supernatant was removed. Further, chloroform (3 ml) was added and washed in the same manner. The residue was dissolved in methanol and then concentrated under reduced pressure, followed by vacuum drying to obtain the title compound (564 mg, 97%) as a pale yellow amorphous substance.
¹ H-NMR (200 MHz, CD ₃ OD): δ 1.44 (3H, s), 1.81 (1H, ddd, J = 13.4, 8.2, 6.2 Hz,), 2.05 ( 3H, s), 2.13 (3H, s), 2.14 (3H, s), 2.17-2.70 (5H, m), 3.06 (1H, ABX system, J _AB = 15. 3 Hz, J _AX = 7.8 Hz), 3.20 (1 H, ABX system, J _AB = 15.3 Hz, J _BX = 5.6 Hz), 3.30-3.48 (2 H, m), 4.66 (1H, dd, J = 7.8, 5.6 Hz), 7.27 (1H, d, J = 1.5 Hz), 8.70 (1H, d, J = 1.5 Hz).
¹³ C-NMR (50.3 MHz, CD ₃ OD): δ 12.0, 12.4, 13.0, 21.6, 24.7, 28.2, 31.0, 36.0, 36.6 , 52.9, 79.1, 118.5, 118.6, 122.2, 123.3, 124.9, 131.5, 135.0, 145.7, 147.3, 173.4, 173 .8, 177.1.
Electrospray ionization mass spectrometry Positive ion mode:
Observation (M + H) ⁺ : 459.1 for C ₂₃ H ₃₁ N ₄ O ₆
Calculated value: 459.2 for isotopic model MH;
Anion mode:
Observation (M-H) ⁻ : 457.1 for C ₂₃ H ₂₉ N ₄ O ₆
Calculated value: 457.2 for isotopic model (M-H).

(Example 3)
1) N-[(2S) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine benzyl ester carnosine benzyl ester dihydrochloride (940 mg, 2.40 mmol) (S) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (601 mg, 2.40 mmol) and dichloromethane (3 ml) were added to a DMF (3 ml) solution at room temperature and dissolved. Subsequently, DIEA (931 mg, 7.20 mmol) and PyBOP (1.25 g, 2.40 mmol) were added, and the reaction solution was stirred at room temperature for 72 hours. After most of dichloromethane was distilled off under reduced pressure, the reaction mixture was diluted with AcOEt (40 ml), and this was washed with water (1 × 30 ml, 3 × 10 ml) and saturated brine (20 ml) (pH of aqueous layer: 5.5). ). The aqueous layer was further extracted with AcOEt (3 × 10 ml), then the entire organic layer was dried (MgSO ₄ ) and concentrated. The oily residue was separated and purified by silica gel column chromatography using chloroform and then chloroform / methanol (15: 1) as a solvent to give the title compound (1.08 g, 81%) as a white amorphous substance.
¹ H-NMR (200 MHz, CDCl ₃ ): δ 1.47 (3H, s), 1.77-1.94 (1H, m), 2.06 (3H, s), 2.13 (6H, s) ), 2.18-2.68 (5H, m), 2.99 and 3.05 (1Heach, ABX system, J _AB = 15.6 Hz, J _AX = 5.7 Hz), 3.37-3. 56 (2H, m), 4.73 (1H, dt, J = 7.5, 5.7 Hz), 5.03 and 5.09 (1Heach, ABq, J = 12.3 Hz), 5.90 ( 2H, br), 6.55 (1H, d, J = 0.9 Hz), 7.11 (1H, d, J = 7.5 Hz), 7.18-7.35 (6H, m), 7. 47 (1H, d, J = 0.9 Hz).

2) N-[(2S) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine compound (3)
10% palladium in a solution of N-[(2S) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine benzyl ester (600 mg, 1.09 mmol) in methanol (3 ml). Carbon (water content: 52.7%) (80 mg) was added, vacuum degassing with an aspirator and hydrogen gas replacement in the reaction flask were repeated 5 times, and then vigorously stirred for 5 hours in a hydrogen atmosphere (using balloons). did. Thereafter, the Pd catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain a yellowish white amorphous. AcOEt (5 ml) was added to this and stirred, and then the supernatant was removed. Further, chloroform (3 ml) was added and washed in the same manner. This amorphous residue was dissolved in methanol and then concentrated under reduced pressure, followed by vacuum drying to obtain the title compound (498 mg, quantitative) as a yellowish white amorphous substance.
¹ H-NMR (200 MHz, CD ₃ OD): δ 1.43 (3H, s), 1.79 (1H, ddd, J = 13.4, 8.2, 6.4 Hz), 2.04 ( 3H, s), 2.12 (3H, s), 2.13 (3H,), 2.18-2.68 (5H, m), 3.02 (1H, ABX system, J _AB = 15.3 Hz. , J _AX = 7.0 Hz), 3.16 (1H, ABX system, J _AB = 15.3 Hz, J _BX = 5.3 Hz), 3.33-3.46 (2H, m), 4.46 ( 1H, dd, J = 7.0, 5.3 Hz), 7.16 (1H, d, J = 1.1 Hz), 8.47 (1H, d, J = 1.1 Hz).
¹³ C-NMR (50.3 MHz, CD ₃ OD): δ 12.0, 12.4, 13.0, 21.6, 24.8, 29.5, 31.0, 36.2, 36.7 , 54.9, 79.1, 118.3, 118.6, 122.3, 123.2, 125.0, 132.8, 134.9, 145.7, 147.3, 173.3, 176 .1, 177.0.
Electrospray ionization mass spectrometry Positive ion mode:
Observation (M + H) ⁺ : 459.1 for C ₂₃ H ₃₁ N ₄ O ₆
Calculated value: 459.2 for isotopic model MH;
Anion mode:
Observation (M-H) ⁻ : 457.1 for C ₂₃ H ₂₉ N ₄ O ₆
Calculated value: 457.2 for isotopic model (M-H).

(Example 4) Antioxidant action (DPPH test)
According to the method of K. Schlesier et al. (Free Radical Research, Vol. 36, 177-187, 2002), 2,2-diphenyl at a concentration of 0.025 mM to 0.150 mM of compounds (1), (2), (3) The scavenging action of the -1-picrylhydrazyl radical (DPPH) was investigated. The results are shown in Table 1 below.
As is clear from Table 1, all of the compounds of the present invention showed DPPH elimination action in a concentration-dependent manner, but L-carnosine showed almost no such action even at high concentrations.

(Example 5) Oxidation inhibiting action of lipoprotein (MDA test)
To 0.5 ml of an aqueous solution containing 105 mM KOH, 20 mM potassium phosphate buffer (pH 7.4), 50 μl of human plasma and 5-25 μl of an aqueous solution of a test compound were added to adjust the final concentrations shown in Table 2 below. Then, 20 μl of a freshly prepared 100 mM FeSO ₄ solution was added and incubated at 37 ° C. for 60 minutes. A 50 μl portion thereof was taken, and malonaldehyde (MDA) produced by lipid peroxidation was quantified as a thiobarbital acid reactive substance (TBRAS) and expressed as a percentage of the control. The results are as shown in Table 2 below.
As is apparent from Table 2, the compounds (1), (2) and (3) of the present invention exhibited a lipoprotein oxidation inhibitory action stronger than that of L-carnosine.

(Example 6) resistance to hydrolytic enzymes human serum 200μl of 125 mM-Tris-HCl buffer _{800μl (pH8.5), 5mM-CdCl} 2 / 30mM- aqueous sodium citrate 300 [mu] l, the mixture of 300 [mu] l of water In addition, after incubation at 37 ° C. for 30 minutes, 400 μl of a 50 mM aqueous test sample solution (adjusted to pH = 8.5 with 0.05 M NaOH) was added to make a final 10 mM solution. This was taken for 0 minutes (before incubation), and 100 μl of each was taken 10, 20, 30, 60, 90, and 180 minutes after the start of incubation, and the remaining amount of the compound was measured using HPLC. The results are as shown in Table 3 below. In the table, the residual ratio of the test compound when 0 minute is taken as 100 is shown. L-carnosine is shown as an average value in serum obtained from two donors 1 and 2, and compounds (2) and (3) are shown as numerical values for each donor.
As is apparent from Table 3, L-carnosine was hydrolyzed relatively rapidly, but the compounds (2) and (3) of the present invention were stable with little degradation even after 90 minutes and 180 minutes.

結果は表４の通りである。なお、表中、紅斑の度合いＫの値はコントロールを１００としたときの値を示す。
表４から明らかなように、本発明化合物はＬ−カルノシン、ＢＨＴよりも優れた紫外線防御作用を示し、特に化合物（３）の作用は最も顕著で、既知の紫外線防御剤であるＢＨＴより有意に優れていた。 (Example 7) UV protection action The UV protection action of compound (2) and compound (3) was measured and compared with control compound L-carnosine and 2,6-di-tert-butyl-4-methylphenol (BHT). did.
Two sites symmetrical with respect to the spine of healthy volunteers (circles each having a diameter of 25 mm) were selected, one of which was coated with a 0.3% solution of the test compound and the other was the control. Both were irradiated with UV light for 1 minute with an HB 400 UV bulb (Philips), and the local erythema intensity I ₀ , I ₂₄ before and 24 hours after irradiation was measured using a sensor Mexometer (Courage & Kazaka Electronics, Germany). The difference ΔI was obtained. The degree K of erythema was determined using the following formula.

The results are shown in Table 4. In the table, the value of the degree K of erythema is a value when the control is 100.
As is apparent from Table 4, the compound of the present invention exhibits an ultraviolet protective effect superior to that of L-carnosine and BHT, and particularly the effect of compound (3) is the most prominent and significantly more significant than BHT, which is a known ultraviolet protective agent. It was excellent.

Example 8 Effect on Moisturizing Action and Elasticity of Skin Epithelium A test area having a diameter of 25 mm was set in the vicinity of the forearm of a healthy person, and a resonator was installed. Here, a 0.3% solution of the test compound was applied at a rate of 50 μl / cm ² , the skin resonance frequency Q before application and 5 minutes after application was measured, and the difference ΔQ was obtained. A larger ΔQ indicates that the skin epithelium swells and increases its elasticity. A significant difference test between ΔQ of each compound was performed using a computer program “Biostatistica”. The results are as shown in Table 5 below.

Results and Discussion L-carnosine was not different from water, but rather tended to have a weak moisturizing effect, which was due to the fact that L-carnosine interacted with water molecules in some way, which was more likely to occur in the skin than with water alone. This is considered to reduce the water content slightly. However, since the compound (3) of the present invention showed a stronger moisturizing effect than water, it was suggested that the compound (3) increases the elasticity of the skin more than the case of L-carnosine aqueous solution or water alone.

The L-carnosine derivative (II) according to the present invention, or a pharmacologically acceptable salt, ester or amide thereof has a stronger antioxidant activity than natural L-carnosine and is resistant to hydrolysis by proteases such as carnosinase. Therefore, when administered orally or parenterally, it can be stably present in various tissues of the living body, and for the prevention and treatment of disorders caused by reactive oxygen species such as human or other mammalian tissues and skin. Useful. Therefore, the target compound of the present invention can be used for the prevention and treatment of ischemic diseases such as arteriosclerosis, myocardial infarction, cerebral infarction, and subarachnoid hemorrhage, and can be expected to have an anti-aging effect, as well as ultraviolet rays. Since it has effects such as a protective action and increased water retention and elasticity of the skin, it is also useful as a cosmetic for improving skin health.

Claims (7)

Formula (II):

Or a pharmacologically acceptable salt, ester or amide thereof.   The compound according to claim 1, wherein the L-carnosine derivative (II) is N-[(2RS) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine.   The compound according to claim 1, wherein the L-carnosine derivative (II) is N-[(2R) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine.   The compound according to claim 1, wherein the L-carnosine derivative (II) is N-[(2S) -6-hydroxy-2,5,7,8-tetramethylchroman-2-carbonyl] -L-carnosine.   The pharmaceutical containing the compound in any one of Claims 1-4.   Cosmetics containing the compound according to any one of claims 1 to 4. The antioxidant containing the compound in any one of Claims 1-4.