JP2011231057A - Ceramide production enhancer - Google Patents

Abstract

（式中、Ｒ_１及びＲ_２はそれぞれ独立に水素原子又は炭素数１〜４のアルキル基を表す。Ｒ_３及びＲ_４は、それぞれ独立に水素原子、水酸基、若しくはメトキシ基を表すか、又はＲ_３とＲ_４はそれらが結合している炭素原子とともに、−ＣＨ＝ＣＨＣ（ＣＨ_３）_２Ｏ−を含む６員環を形成する。）
【選択図】なしA ceramide production promoter is provided.
A ceramide production promoter comprising a compound represented by the following general formula (1) as an active ingredient.

(In the formula, R ₁ and R ₂ each independently represent a hydrogen atom or an alkyl group having 1 to 4 carbon atoms. R ₃ and R ₄ each independently represent a hydrogen atom, a hydroxyl group, or a methoxy group, or R ₃ and R ₄ together with the carbon atom to which they are attached form a 6-membered ring containing —CH═CHC (CH ₃ ) ₂ O—.
[Selection figure] None

Description

  The present invention relates to a ceramide production promoter.

Ceramide is a component of sphingolipid and is present in a wide range in a small amount in animal and plant tissues.
In recent years, it has been reported that ceramide has bone resorption inhibitory action, bone strengthening action, and alveolar bone loss inhibitory action, and is useful for the prevention and improvement of bone and joint diseases such as osteoporosis, fracture, low back pain, rheumatism ( Patent Document 1) has been reported. In addition, it has been reported that ceramide has an effect of imparting firmness and stiffness to the hair and improving touch (see Patent Document 2).
Attempts have been made to supply ceramide from the outside in order to keep the amount of ceramide in the tissue normal, but the effect is not always sufficient at present.

JP 2001-158736 A Japanese Patent Laid-Open No. 10-152421

  An object of the present invention is to provide a ceramide production promoter having a high ceramide production promotion effect.

  In view of the above problems, the present inventors have intensively studied to search for a novel substance having a ceramide production promoting action. As a result, it was found that the compound represented by the following general formula (1) has a high ceramide production promoting action. The present invention has been made based on this finding.

  That is, this invention relates to the ceramide production promoter which contains the compound represented by following General formula (1) as an active ingredient.

In General Formula (1), R ₁ and R ₂ each independently represent a hydrogen atom or an alkyl group having 1 to 4 carbon atoms. R ₃ and R ₄ each independently represent a hydrogen atom, a hydroxyl group, or a methoxy group, or R ₃ and R ₄ together with the carbon atom to which they are bonded together, —CH═CHC (CH ₃ ) ₂ O— To form a 6-membered ring.

  According to the present invention, a ceramide production promoter can be provided. The ceramide production promoter of the present invention can supply ceramide from the outside, for example, in the form of an external preparation for skin, and can maintain the amount of ceramide in the tissue within a normal range or compensate for the decreased amount of ceramide.

Hereinafter, the present invention will be described in detail.
The ceramide production promoter of the present invention contains a compound represented by the following general formula (1) as an active ingredient. As demonstrated in Examples described later, this compound has a high ceramide production promoting action.

In General Formula (1), R ₁ and R ₂ each independently represent a hydrogen atom or an alkyl group having 1 to 4 carbon atoms. Specific examples of the alkyl group having 1 to 4 carbon atoms include a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, and an isobutyl group. Of these, R ₁ is preferably a hydrogen atom or a methyl group. R ₂ is preferably a hydrogen atom or a methyl group.
In general formula (1), R ₃ and R ₄ each independently represent a hydrogen atom, a hydroxyl group, or a methoxy group, or R ₃ and R ₄ together with the carbon atom to which they are bonded together, —CH═CHC A 6-membered ring containing (CH ₃ ) ₂ O— is formed. Among these, R ₃ is preferably a hydroxyl group or a methoxy group, and more preferably a hydroxyl group. R ₄ is preferably a hydrogen atom. It is also preferable that R ₃ and R ₄ together with the carbon atom to which R ₃ and R ₄ are bonded form a 6-membered ring containing —CH═CHC (CH ₃ ) ₂ O—.

  In the present invention, the compound represented by the general formula (1) includes a salt thereof. Examples of the salt include, but are not limited to, alkali metal salts such as lithium salt, sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, alkylamine salts such as trimethylamine and triethylamine, and quaternary ammonium salts. , Alkanolamine salts such as triethanolamine, diethanolamine and monoethanolamine, or amino acid salts such as lysine, histidine and arginine.

  Although the specific example of a compound represented by General formula (1) below is given, this invention is not limited by these. In the following exemplary compounds, Me represents a methyl group.

The compound represented by the general formula (1) can be produced by chemical synthesis. For example, it can be synthesized from various aromatic rings as raw materials through addition reaction, cyclization reaction, alkylation reaction and the like. Specifically, the compound can be synthesized with reference to the synthesis methods described in Phytochemistry, 19, 921, 1980, Australian Journal of Chemistry, 40, 1705, 1987, JP-A-2005-133272, and the like.
As a more specific synthesis method, the method shown in the Examples described later can be used, but the present invention is not limited to this.

  Moreover, the compound represented by the said General formula (1) can also be obtained from what is marketed as a reagent etc. For example, it is available from Wako Pure Chemical Industries, Kanto Chemical Co., Funakoshi Co., Ltd., ALDRICH Co., etc.

  Furthermore, the compound represented by the general formula (1) can also be obtained by extraction and isolation from plant materials. The method for isolating the compound represented by the general formula (1) is not particularly limited. For example, the plant is extracted using an appropriate solvent, and the obtained plant extract is subjected to a technique such as chromatography. The method of isolating the compound represented by General formula (1) is mentioned.

The compound represented by the general formula (1) has an excellent ceramide production promoting action as shown in Examples described later, and a ceramide production promoter can be obtained by containing this compound. As described above, ceramide has been reported to have bone resorption inhibitory action, bone strengthening action, and alveolar bone loss inhibitory action, and the ceramide production promoter of the present invention is effective for bones such as osteoporosis, fractures, low back pain, and rheumatism. It is useful as a material for preventing or ameliorating joint diseases, preventing or improving periodontal diseases, pharmaceuticals, quasi drugs and the like. In addition, it has been reported that ceramide has an effect of imparting elasticity and feel to hair, and the ceramide production promoter of the present invention imparts elasticity and feel to hair and improves the feel of hair. It is also useful as a raw material for quasi-drugs and cosmetics.
It has never been known that the compound represented by the general formula (1) has a ceramide production promoting action, which is a new finding obtained by the present inventors.

In the present invention, the compound represented by the general formula (1) may be used as it is as a ceramide production promoter. Or you may add various additives etc. to the compound represented by the said General formula (1) in the range which does not affect the effect. For example, an appropriate liquid or solid excipient or filler such as titanium oxide, calcium carbonate, distilled water, lactose, starch, etc. may be added and used as a ceramide production promoter.
When the composition is used, the amount of the compound represented by the general formula (1) in the ceramide production promoter is not particularly limited, but the compound represented by the general formula (1) is 0.00001 to 20% by mass. It is preferably contained, and particularly preferably about 0.0001 to 10% by mass.

The ceramide production promoter of the present invention can be applied to cosmetics, quasi drugs, and pharmaceuticals.
When used for cosmetics or quasi drugs, for example, it can be in the form of a skin external preparation. When used in the form of an external preparation for skin, in addition to the compound represented by the general formula (1), the above-mentioned various additives and other medicinal components can be appropriately added, and further, depending on the dosage form that can be taken. Various components that are usually used in external preparations for skin can be blended. Specific examples of dosage forms for external preparations for skin include various dosage forms that can be applied externally, such as creams, emulsions, lotions, gels, ointments, pastes, packs, sheet-like products, and the like. For example, various oil agents, surfactants, gelling agents, preservatives, antioxidants, solvents, alcohol, water, chelating agents, thickeners, ultraviolet absorbers, emulsion stabilizers, pH adjusters, dyes, and fragrances Etc. can be blended. In addition, other medicinal ingredients can be blended. Examples of such medicinal ingredients include other moisturizing ingredients such as sodium hyaluronate, known ceramide production promoters, pseudoceramides, natural ceramides, sugar ceramides, sphingomyelin, skin anti-aging agents, and whitening agents. .

Known ceramide production promoters are not particularly limited, and examples thereof include acetylhydroxyproline, potassium glycyrrhizinate, L-carnitine, ascorbic acid, ascorbyl glucoside, magnesium ascorbyl phosphate, dl-α-tocopheryl-dl. -Ascorbyl phosphate, dl-α-tocopheryl phosphate, nicotinamide, tocopherol nicotinate, L-lactic acid, vitamin C, asparagus extract, butcher bloom, genquanine, rosemary, lavender, sage, jujube, black ( Red) Ganoderma, Toki, Kujin, Yokuinin, Beniceanne extract, Rice power extract, etc.
The pseudo-ceramide is not particularly limited. For example, commercially available ceramide R (manufactured by Unilever), ceramide PC-104 (manufactured by Taiheiyo Chemical), ceramide HO3 (manufactured by sederma), Erduo PS-203 (manufactured by Ajinomoto) Etc.
Examples of sugar ceramides include, but are not limited to, glucosyl ceramide, galactosyl ceramide, and the like, but commercially available products include nippon ceramide (manufactured by Nippon Flour), oriza ceramide (manufactured by Oriza Yuka), Nissan ceramide. , Neo Liquid Ceramide N (manufactured by Nippon Oil & Fats), Ceramide (manufactured by Unitika) and the like.
The content of the compound represented by the general formula (1) in the external preparation for skin such as cosmetics is preferably 0.00001 to 20% by mass, particularly 0.0001 to 10% by mass. preferable.

  Examples of the dosage form when the ceramide production promoter of the present invention is used as a pharmaceutical include oral administration by tablets, capsules, granules, powders, syrups, etc. or injections, external preparations, suppositories, transdermal absorption agents, etc. Parenteral administration. To prepare the pharmaceutical preparation, the ceramide production promoter of the present invention alone or other pharmaceutically acceptable excipients, binders, extenders, disintegrants, surfactants, lubricants, A dispersant, buffer, preservative, flavoring agent, fragrance, coating agent, carrier, diluent and the like can be used in appropriate combination. The content of the compound represented by the general formula (1) in the preparation is preferably 0.00001 to 20% by mass, particularly preferably 0.0001 to 10% by mass. When the ceramide production promoter of the present invention is used as a pharmaceutical, the daily dose per adult is preferably 0.001 to 1000 mg of the compound represented by the general formula (1), particularly The amount is preferably 0.01 to 100 mg.

  EXAMPLES Hereinafter, although this invention is demonstrated further in detail based on an Example, this invention is not limited to this.

Synthesis Example The exemplified compounds (b) and (c) were respectively synthesized by the following methods.

1. Exemplary Compound (b): Synthesis of 4′-O-methylgrabridine 300 mg (0.925 mmol) of glabrizine was added to a 50 mL eggplant flask and dissolved in 13.5 mL of chloroform and 1.5 mL of methanol. Under ice cooling, 5.55 mL (11.1 mmol) of a trimethylsilyldiazomethane 2.0M hexane solution was slowly added dropwise and stirred for 1 hour, and then stirred at room temperature for 47 hours. Under ice-cooling, 69.9 μL (13.3 mmol) of acetic acid was added to stop the reaction. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography 50 g. As a colorless solid, 111 mg of 4′-O-methylgrabridine (exemplary compound (b)) (yield 35.4%) and as a colorless solid grabridine- 118 mg (yield 36.6%) of dimethyl compound was obtained.

The structure of the obtained compound was confirmed by ¹ H-NMR and ¹³ C-NMR.
¹ H-NMR (CDCl _3, 600 MHz) δ1.34 (3H, s), 1.36 (3H, s), 2.79 (1H, ddd, J = 15.6 Hz, 5.2 Hz, 2.1 Hz), 2.92 (1H, ddd , J = 15.6Hz, 10.9Hz, 0.8Hz), 3.42 (1H, dddd, J = 10.9Hz, 10.3Hz, 5.2Hz, 3.4Hz), 3.69 (3H, s), 3.95 (1H, ddd, J = 10.3) Hz, 10.3Hz, 0.8Hz), 4.31 (1H, ddd, J = 10.3Hz, 3.4Hz, 2.1Hz), 4.90 (1H, br), 5.49 (1H, d, J = 9.9Hz), 6.29 (1H, d, J = 2.5Hz), 6.30 (1H, d, J = 8.5Hz), 6.41 (1H, dd, J = 8.3Hz, 2.5Hz), 6.58 (1H, dd, J = 9.9Hz, 0.6Hz), 6.76 (1H, br-d, J = 8.3Hz), 6.95 (1H, d, J = 8.5Hz)
¹³ C-NMR (CDCl ₃ ,, 125 MHz) δ 27.54, 27.78, 30.59, 31.68, 55.36, 70.01, 75.62, 96.14, 103.13, 106.00, 108.71, 109.93, 114.36, 116.95, 119.85, 128.22, 128.98, 129.21, 149.74 , 151.87, 154.26, 159.35

2. Exemplary Compound (c): Synthesis of Satiban
Synthesis of Intermediate 1 (1- (4- (benzyloxy) -2-hydroxyphenyl) ethanone) 2, 10 g (65.7 mmol) of 4 -dihydroxyacetophenone was added to a 1 mL eggplant flask under a nitrogen atmosphere and dissolved in 400 mL of acetonitrile. . 9.51 mL (85.4 mmol) of benzyl chloride, 685 mg (6.57 mmol) of sodium iodide and 12.1 g (144.5 mmol) of sodium hydrogen carbonate were added, and the mixture was stirred at 50 ° C. for 4 days. The reaction solution was filtered through celite, and the solvent was evaporated under reduced pressure. Ethyl acetate 200mL and water 200mL were added, liquid separation operation was performed, the organic layer was extracted, and the solvent was distilled off under reduced pressure. Hexane 150mL and ethyl acetate 15mL were added, it was made to melt | dissolve at 60 degreeC, and it air-cooled, stirring. The precipitated crystals were filtered (φ = 9.0 cm, filter paper 5B) to obtain Intermediate 1. (Yield 12.7 g, Yield 78.5%)

Intermediate 2 (3- (2,4-bis (methoxy) phenyl) -7- (benzyloxy) -4H-chromen-4-one) Synthetic Intermediate 1 1.50 g (6.19 mmol), 2,4- Dimethoxybenzaldehyde (1.03 g, 6.19 mmol) and ethanol (50 mL) were added to a 100 mL eggplant flask, and the mixture was heated to 80 ° C. and stirred. 8.3 ml of 50% aqueous sodium hydroxide solution was gradually added dropwise and refluxed for 30 minutes. The reaction was stopped by adding 1M hydrochloric acid until pH acidic. The precipitate was filtered (φ = 9.0 cm, filter paper 5A), washed with water, and dried under reduced pressure. 20 mL of hexane and 20 mL of ethyl acetate were added, dissolved at 60 ° C., and air-cooled with stirring. The precipitated crystals were filtered (φ = 6.0 cm, filter paper 5A) and dried under reduced pressure. 980 mg of dried solid was added to a 500 mL eggplant flask, dissolved in 300 mL of ethanol, and heated to 50 ° C. After adding 1.34 g (3.01 mmol) of thallium (III) nitrate trihydrate and stirring for 3 hours, 9 ml of 3N hydrochloric acid was added and further stirred for 12 hours. The precipitate was filtered (φ = 6.0 cm, filter paper 5B), and the filtrate was evaporated under reduced pressure. Purification by silica gel column chromatography 45 g (chloroform → chloroform: methanol = 49: 1) gave Intermediate 2. (Yield 487 mg, Yield 41.2%)

300 mg (0.772 mmol) of synthetic intermediate 2 of Satiban (3,4-dihydro-3- (2,4-dimethoxyphenyl) -2H-chromen-7-ol) was added to a 200 mL eggplant flask, 30 mL of acetone, 1. Dissolved in 5 mL. 10% palladium-carbon (300 mg) was added, and the mixture was stirred at 40 ° C. for 6 hours under a hydrogen atmosphere. The reaction solution was filtered through Celite, and the filtrate was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. Purification by silica gel column chromatography 45 g (chloroform → chloroform: methanol = 49: 1) gave a sativan (exemplary compound (c)) as a colorless solid. (Yield 172 mg, Yield 81.12%)

The structure of the obtained compound was confirmed by ¹ H-NMR and ¹³ C-NMR.
¹ H-NMR (CDCl ₃ ,, 600 MHz) δ 2.86 (1 H, ddd, J = 15.7 Hz, 5.5 Hz, 2.0 Hz), 2.97 (1 H, ddd, J = 15.7 Hz, 10.7 Hz, 0.5 Hz), 3.56 (1H, dddd, J = 10.7Hz, 10.3Hz, 5.5Hz, 3.6Hz), 3.80 (3H, s), 3.81 (3H, s), 3.99 (1H, dd, J = 10.3Hz, 10.3Hz), 4.29 (1H, ddd, J = 10.3Hz, 3.6Hz, 2.0Hz), 4.83 (1H, br), 6.35 (1H, d, J = 2.5Hz), 6.38 (1H, dd, J = 8.3Hz, 2.5Hz) , 6.46 (1H, dd, J = 8.3Hz, 2.5Hz), 6.49 (1H, d, J = 2.5Hz), 6.94 (1H, d, J = 8.3Hz), 7.02 (1H, d, J = 8.3Hz) )
¹³ C-NMR (CDCl ₃ ,, 125 MHz) δ30.33, 31.48, 55.34, 55.37, 70.11, 98.67, 103.16, 104.04, 107.81, 114.86, 121.79, 127.53, 130.41, 154.78, 155.17, 158.26, 159.63

(Test example) Verification of ceramide production promoting effect Using the exemplified compounds (a) to (c), the ceramide production promoting action was evaluated by the following procedure. Exemplified compound (b) 4'-O-methylgrabridine and (c) sativa are those synthesized above, and exemplary compound (a) grabridine is commercially available from Wako Pure Chemicals as a reagent. used.

Using a culture plate, normal human epidermal keratinocytes (trade name: NHEK (F), manufactured by KURABO) are cultured at 37 ° C., 5% CO _{2 in} a culture solution (trade name: EpiLife-KG2, manufactured by KURABO). In culture.
Thereafter, the culture solution is replaced with EpiLife-KG2 excluding growth factors such as epidermal growth factor, and the concentrations of the evaluation samples (exemplary compounds (a) to (c), comparative compounds) are 10 mM, 5 mM, 2 mM, and 1 mM. As described above, 0.1% amount of each prepared or control solution (50% ethanol) was added.
After culturing for 3 days, each cell was collected together with 1 well.

The organic phase from which the lipid was extracted from the collected cells by the Bligh and Dyer method was transferred to a glass tube, solidified with nitrogen, and then redissolved with chloroform and methanol to obtain a lipid sample.
In addition, 0.1N NaOH, 1% SDS aqueous solution is added to the cells after lipid extraction, and the protein is solubilized by heating at 60 ° C. for 2 hours, cooled to room temperature, and then neutralized by adding 2N HCl. The amount of protein was quantified by the BCA method.

The prepared lipid sample was horizontally developed by thin film chromatography (TLC) twice with chloroform: methanol: acetic acid = 190: 9: 1. The copper sulfate solution was sprayed and sprayed with a hot plate to detect ceramide, and the amount was determined.
The results are shown in Table 1. In addition, the amount of ceramide shown in Table 1 shows a relative value when the control solution addition group is 1.

  As a comparative compound, the same evaluation as described above was performed using isoliquiritigenin (manufactured by Funakoshi) having the following structure.

  As is apparent from Table 1, it was recognized that the ceramide yield was increased in the system to which the exemplary compounds (a) to (c) were added as compared with the control system. Therefore, it turns out that the ceramide production promoter of this invention which contains the compound represented by the said General formula (1) as an active ingredient can accelerate | stimulate ceramide production.

(Prescription Example 1)
Hair lotions (Nos. 1 to 3) having the compositions shown in Table 2 below were produced by conventional methods.

(Prescription example 2)
Hair treatments (Nos. 11 to 13) having the compositions shown in Table 3 below were produced by conventional methods.

Claims (1)

The ceramide production promoter which contains the compound represented by following General formula (1) as an active ingredient.

(In the formula, R ₁ and R ₂ each independently represent a hydrogen atom or an alkyl group having 1 to 4 carbon atoms. R ₃ and R ₄ each independently represent a hydrogen atom, a hydroxyl group, or a methoxy group, or R ₃ and R ₄ together with the carbon atom to which they are attached form a 6-membered ring containing —CH═CHC (CH ₃ ) ₂ O—.