JP2003119150A - Lipase inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a lipase inhibitor having high safety, with which pimples, rough skin, dermatitis, diaper rash and the like can be suppressed. SOLUTION: The lipase inhibitor comprises solvent extracts of one or more kinds selected from Eucalyptus plants as active ingredient. As the solvent, a lower alcohol, a glycol, a halogenated hydrocarbon, an ether, a lower fatty acid ester, a ketone, water or the like is used. Among extracted components, a dihydrochalcone has a lipase inhibiting activity.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a lipase inhibitor.

[0002]

2. Description of the Related Art The sebum present on the skin surface is decomposed into fatty acids by lipase produced by bacteria, and the barrier ability of the skin is reduced. As a result, the stimulant easily penetrates into the skin, which induces inflammation and causes acne and rough skin. In addition, the produced fatty acid and the oxidative decomposition product of the fatty acid cause a bad odor. Lipase is also involved in the development of dermatitis such as diaper rash, and it is considered that the action of lipase present in feces on the surface of the skin lowers the barrier ability of the skin and induces inflammation. Therefore, with lipase inhibitors, acne, rough skin, dermatitis,
It is thought that diaper rash etc. can be suppressed and prevented. The lipid contained in the diet is decomposed by lipase in the pancreas and absorbed from the small intestine. Excessive intake of lipids leads to obesity and causes hyperlipidemia, arteriosclerosis, heart disease, diabetes and the like. If a lipase inhibitor can suppress the accumulation of fat in the body, it is considered that obesity can be suppressed or prevented.

Conventionally, protein substances in soybean seeds (Agric. Biol. Chem., 38, 97, 197) have been used as lipase inhibitors.
4), serum albumin (Gastroenterology, 75, 382, 19
78), β-lactoglobulin (J. Clin. Invest., 64,
1303, 1979) and other proteins and Triton X-100 (Biochimie, 5
8, 751, 1976) and bile salts (Z. Physiol. Chem., 125, 1
32, 1923) and other surfactants, etc. have been reported. However, with proteins, there are problems with stability during storage,
When used on human skin, there is concern about the development of allergies. Further, surfactants generally have strong skin irritation, and there is concern about safety when used on the skin. Therefore, when using it on human skin, etc., or when orally administered,
There is a demand for inhibitors that are highly safe for the human body. Among plant extracts, extracts such as rosemary (JP-A-10-26260
No. 6), extracts such as Asenyaku (JP 2000-103741A), etc., but there are reported cases of eucalyptus extracts that have been confirmed to be safe for food additives and cosmetics. I can't find it.

[0004]

DISCLOSURE OF THE INVENTION The present invention is to provide a highly safe lipase inhibitor.

[0005]

In order to solve the above problems, the present invention adopts the following configurations. That is, the present invention is
“A lipase inhibitor containing a solvent extract of one or more plant species selected from the genus Eucalyptus as an active ingredient”.

In the present invention, the solvent used for extraction is one kind of solvent selected from lower alcohols, glycols, halogenated hydrocarbons, ethers, lower fatty acid esters, ketones and water, or these solvents. Among them, any mixed solvent using two or more kinds of uniformly mixable solvents is preferable.

In the above-mentioned present invention, Eucalyptus plants include E. globulus , E. viminalis , E. camaldulen.
It is preferably selected from sis , E. grandis and E. nitens .

Examples of the solvent extract of the Eucalyptus plant include an extract containing a dihydrochalcone compound represented by the following chemical formula (1) as a main active ingredient.

[Chemical 3]

Further, the dihydrochalcone represented by the above chemical formula (1) can be used as a lipase inhibitor by itself.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION The extract used in the present invention is an extract obtained by using leaves of a eucalyptus plant as a medium to be extracted and extracting this with a commonly used organic solvent, water, or an arbitrary mixed solvent thereof. Is. Examples of the organic solvent include lower alcohols such as ethanol, methanol and propanol, glycols such as propylene glycol and butylene glycol, halogenated hydrocarbons such as chloroform, dichloromethane and dichloroethane, methyl ether, ethyl ether and dioxane. Examples thereof include lower fatty acid esters such as ethers, methyl acetate, ethyl acetate and butyl acetate, and ketones such as acetone and methyl ethyl ketone. These solvents may be used alone or as a mixed solvent of any two or more kinds.

With regard to the genus Eucalyptus, any plant species belonging to the same genus can be used, and may be used alone or in combination of two or more species. For example,
Eucalyptus globulus , E. niten
s , E. camaldulensis , E. viminalis , E. grandis, etc. can be used. The parts to be extracted are leaves, stems,
Any part such as a trunk or a root may be used, but leaves are preferable from the viewpoint of extraction efficiency.

The extraction method may be a generally used method, for example, a method of immersing the raw material in a solvent for a long time,
There is a method in which extraction is carried out while heating and stirring at a temperature not higher than the boiling point of the organic solvent, followed by filtration to obtain an extract. The obtained extract may be used as it is, or may be concentrated by vacuum concentration and used. As a pre-extraction step, the raw material may be defatted with a non-polar solvent to remove the essential oil and then extracted with the solvent, or may be extracted with the solvent after removing the essential oil by steam distillation. The obtained extract may be used after being treated with an adsorbent such as activated carbon, celite or silica gel to remove color and odor.

As shown in the production examples below, a leaf extract of Eucalyptus globulus contains a dihydrochalcone compound represented by the above chemical formula (1) (JP-A-11-80012) as a compound having a lipase inhibitory activity. Has been. The compound having a lipase inhibitory activity contained in eucalyptus leaves is considered to include compounds other than these compounds, but when preparing the eucalyptus extract used in the present invention, these compounds or their analogs Can be used as an index.

When the extract obtained from the above plant is used as a lipase inhibitor, the dosage form is not particularly limited, and it can be used in the form of liquid, gel, powder or the like.
In addition, the extract can be blended with lotions, sprays, mousses and the like. In the case of a liquid or gel product, the extract is preferably contained in an amount of 0.00001 to 10% by weight, more preferably 0.001 to 1% by weight.

The extract extracted by the above method can be used as a lipase inhibitor on human or animal skin. Further, it can be contained in diapers, tissues, wipes for wipes, hygiene sheets, nursing sheets, etc., or can be used as a wet tissue medicine, cleaning agent, etc. When contained in a diaper, for example, a liquid composition is applied to a diaper (cloth, paper, non-woven fabric, etc.), or impregnated and dried, a powdered composition is mixed with the diaper, and the composition is made into a lotion. It can be carried out by a method such as blending and applying lotion to the diaper. In addition, it may be added to cosmetics such as creams and lotions to prevent and suppress acne, rough skin, dermatitis such as diaper rash, and beverages, foods, tablets, capsules, etc., or to be used as a health food or pharmaceutical raw material. You can also

When the above extract is used as a lipase inhibitor, various components, moisturizers and acidity regulators commonly used in foods, pharmaceuticals, quasi drugs, cosmetics, etc. are used within a range that does not impair the effects of the present invention. , Stabilizers, surfactants, antioxidants, deodorants, antibacterial agents, etc. can be blended, and these auxiliary components may be blended in two or more components.

Various ingredients generally used in foods, pharmaceuticals, quasi drugs, cosmetics and the like include allergy suppressants, atopic dermatitis suppressors, itch suppressors, skin roughening suppressors, lower alcohols and polyhydric alcohols. , Fragrances, cooling agents, animal and plant polysaccharides and degradation products thereof, animal and plant glycoproteins and degradation products thereof, microbial culture metabolic components, amino acids and salts thereof, deodorants, emulsifiers, etc., and can be used in combination .

The moisturizers include aloe extract, protracted herb extract, hypericum extract, barley extract, orange extract, seaweed extract, chamomile extract, cucumber extract, comfrey extract, burdock extract, shiitake extract, perilla extract, perilla extract, sage extract, Duke extract, Cordyceps sinensis extract, Dokudami extract, Hatake shimeji extract, loquat extract, grape leaf extract, fuyubodaiju extract, prune extract, loofah extract, button pi extract, makai extract, momoha extract, lily extract, apple extract, almond oil, olive oil, sesame oil, safflower Oil, dimethyl silicone, silicone oil, modified silicone, soybean oil, camellia oil, castor oil, jojoba oil, mink oil, coconut oil, arabinose, galactose,
Xylose, glucose, sucrose, sorbitol, fructose, maltose, maltitol, mannose, beeswax, hyaluronic acid, placenta extract, rhamnose, xylobiose, xylooligosaccharide, tuberose polysaccharide, trisaccharide, trehalose,
Ethylene glycol, soluble collagen, glycerin,
Glycyrrhizin, chondroitin sulfate, diglycerin,
Examples thereof include squalane, ceramide-like compounds, triglycerin, urea, vitamin C phosphate calcium salt, vitamin E, sodium pyrrolidonecarboxylate, hinokitiol, 1,3-butylene glycol, propylene glycol, liquid paraffin and petrolatum.

Among these moisturizers, aloe extract, prolong life herb extract, hypericum extract, comfrey extract,
Perilla extract, sage extract, ceramide-like compound, Dokudami extract, Hatake shimeji extract, loquat extract, Fujudaiju extract, button pi extract, castor oil, jojoba oil, lanolin, hyaluronic acid, placenta extract, rhamnose, xylooligosaccharide, soluble collagen, glycerin, chondroitin Sulfuric acid, squalane, urea, 1,3
-Butylene glycol and propylene glycol are more preferred, and aloe extract, xylooligosaccharides, ceramide-like compounds, edible mushroom extract, urea, 1,3-butylene glycol and propylene glycol are particularly preferred.

The acidity regulator and the function stabilizer include:
For example, adipic acid, citric acid, trisodium citrate, glycine, glycerin fatty acid ester, gluconodelta lactone, gluconic acid, succinic acid, monosodium succinate, disodium succinate, acetic acid, sodium acetate, DL-tartaric acid, L -Tartaric acid, DL-sodium tartrate, L-sodium tartrate, carbonates, carbon dioxide, lactic acid, sodium lactate, fumaric acid, monosodium fumarate, lysozyme, DL-malic acid, DL-sodium malate, phosphoric acid, phosphoric acid Examples thereof include salts, polymerized phosphates, itaconic acid, phytic acid and the like.

Examples of the surfactant include glycerin fatty acid esters such as glycerol monostearate and polyglycerol trioleate, organic acid monoglycerides,
Propylene glycol fatty acid ester, sorbitan fatty acid ester, sucrose fatty acid ester, lecithin, lysolecithin, polyethylene glycol, polyoxyalkyl ether, polyoxyethylene polyamine, alkyl polyoxyethylene sulfuric acid ester salt, alkyl sulfuric acid ester salt, acylmethyl taurine salt, N -Acyl glutamate, alkylamido betaine and the like can be mentioned.

Examples of antioxidants include catechin, tocopherol, propolis, ellagic acid, and animal and plant extracts (sage, seri, rosemary, etc.).

The deodorant is not particularly limited as long as it is a substance or an extract having a deodorizing activity, and examples thereof include extracts of plants and mushrooms, essential oils and the like.

The antibacterial agent is not particularly limited as long as it is a substance or extract having antibacterial activity, and examples thereof include hinokitiol, triclosan, cetylpyridinium chloride, chlorhexidine gluconate, potassium sorbate, sorbic acid, sodium benzoate, and benzoic acid. Examples thereof include acids, paraoxybenzoic acids such as ethyl paraoxybenzoate and propyl paraoxybenzoate, animal / plant extracts, essential oils and the like.

Hereinafter, the present invention will be described in detail with reference to Examples, but these Examples are illustrative and the scope of the present invention is defined by the scope of the claims.

<Production Example 1> Eucalyptus glo of Eucalyptus
bulus , E. nitens , E. camaldulensis , E. viminalis ,
E. grandis ) 50 g of each leaf is mixed with 50% ethanol (1
L) was extracted for 3 days at room temperature to obtain each extract. These extracts were concentrated under reduced pressure to obtain an extract.

<Production Example 2> Eucalyptus glo
bulus , E. nitens , E. camaldulensis , E. viminalis ,
20 g of leaves of E. grandis ) were extracted with 50% butylene glycol (160 ml) for 3 days at room temperature to obtain an extract.

<Production Example 3> Eucalyptus ( E. globulus )
Leaf 500 g was extracted with 50% ethanol (7 L) for 3 days at room temperature to obtain an extract. Concentrate the extract under reduced pressure,
98 g (19.6% yield from leaves) of the extract was obtained.

Next, the extract of Production Example 3 was partitioned and extracted with hexane and water, the hexane layer was separated, and then the aqueous layer was partitioned and extracted with dichloromethane, ethyl acetate and n-butanol in that order. Concentrate the ethyl acetate fraction as a highly active fraction,
10.24 g of an ethyl acetate extract was obtained (2.0% yield from leaves).

After the ethyl acetate extract was adsorbed on silica gel chromatography, it was eluted with a hexane-ethyl acetate mixed solvent for fractionation. The fraction eluted with a hexane / ethyl acetate = 1: 4 mixture was concentrated to obtain 6.24 g of an active fraction (yield 1.2% from leaves). This fraction was further separated by ODS-HPLC (solvent: 80% methanol solution) to give compound (1) 0.4
25 g was obtained (content in leaf was 0.085%).

The compound (1) was found to be dihydrochalcone represented by the above chemical formula (1) from the following physicochemical properties. Molecular weight: EI-MS m / z 286 (M +), 181,
154, molecular formula: C17H18O4 (λmax MeOH): 286nm (ε = 2170
0), IR (νmax KBr): 3296, 2944, 2924, 1650, 1595,
1516, 1429, 1274, 1247, 1213, 1147, 1112, 1082, 88
6, 799, 742, 720, 699, 468cm-1 1H-NMR (δdmso-d6): 1.86 (3H, s), 2.89 (1H, t, J =
7.6), 3.32 (1H, t, J = 7.6), 3.77 (3H, s), 6.08 (1H, s),
7.15-7.30 (5H, m), 10.92 (1H, s), 13.64 (1H, s) ppm 13C-NMR (δ dmso-d6): 7.2, 30.1, 45.3, 55.4, 90.
3, 102.3, 104.1, 125.8, 128.3, 141.6, 160.5, 162.
0, 163.2, 204.7 ppm

<Example 1> Each of the extracts obtained in Production Example 1 was dissolved in dimethyl sulfoxide (Wako Pure Chemical Industries, Ltd.) so that the concentration of the extract was 1%, and a sample solution was prepared. The activity was measured. As a reagent, lipase (Si
gma; TypeII, Crude, from Porcine pancreas), Lipase Kit S manufactured by Dainippon Pharmaceutical. The lipase was dissolved in 10 mM phosphate buffer pH 8.0. Sample solution 5
0 μl, coloring solution (buffer containing 0.1 mg / ml 5,5-diobis)
Add 780 μl and 20 μl of esterase inhibitor solution (3.48 mg / ml phenylmethylsulfonyl chloride), and add 5 at 30 ℃.
After heating for minutes, 100 μl of the substrate solution (6.69 mg / ml methylcapnol tributyrate + 5.73 mg / ml sodium dodecyl sulfate) and 50 μl of lipase solution were added, and the mixture was incubated at 30 ° C. for 30 minutes in the dark. Add 2 ml of reaction stop solution and
The absorbance at 12 nm was measured to determine the inhibition rate (%). The results are shown in Table 1, and the lipase inhibitory activity was observed in each eucalyptus extract.

<Comparative Example 1> Camphor tree (Cinnamomum camph)
ore), Stellax japonica, Tobera (Pittos)
50g of porum tobira leaves with 50% ethanol (1L)
Extraction was performed for 3 days at room temperature. This extract was concentrated under reduced pressure to obtain an extract. For each extract, a dimethylsulfoxide solution having an extract concentration of 1% was prepared as a sample solution, and the lipase inhibitory activity was measured in the same manner as in Example 1 below. The results are shown in Table 1, but no inhibitory activity was observed.

[0034]

[Table 1]

Example 2 The extract solution (stock solution) obtained in Production Example 2 was diluted with butylene glycol to obtain an extract concentration of 1
% Solution was made. Using this as a sample solution, the results of measuring the lipase inhibitory activity by the method described in Example 1 are shown in Table 2 below. A lipase inhibitory activity was observed in each eucalyptus extract.

<Comparative Example 2> Camphor tree (Cinnamomum camph)
ore), Stellax japonica, Tobera (Pittos)
20 g of leaves of porum tobira) were extracted with 50% butylene glycol (160 ml) for 3 days at room temperature to obtain an extract. The lipase inhibitory activity of each extract was measured in the same manner as in Example 2. The results are shown in Table 2, but no inhibitory activity was observed.

[0037]

[Table 2]

Example 3 The results of measuring the lipase inhibitory activity of the extract obtained in Production Example 3 in the same manner as in Example 1 are shown in Table 3. Further, the compound (1) obtained in Production Example 3 was dissolved in dimethylsulfoxide (Wako Pure Chemical Industries, Ltd.) so that the concentration was 1% to prepare a sample solution, and lipase inhibitory activity was carried out in the same manner as in Example 1. Was measured. The results are shown in Table 3.

[0039]

[Table 3]

[0040]

EFFECTS OF THE INVENTION From Tables 1 and 2, it is clear that the extract of Eucalyptus obtained in the present invention has a sufficient lipase inhibitor, and therefore contains a lipase inhibitor. Furthermore, from Table 3, it was also confirmed that the dihydrochalcone represented by the chemical formula (1) has a lipase inhibitory activity. Further, the lipase inhibitor obtained in the present invention is derived from a natural product and has high safety.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 17/00 A61P 17/00 43/00 111 43/00 111 C12N 9/99 C12N 9/99 F term ( Reference) 4B018 MD08 MD61 ME14 MF01 4C083 AA111 AA112 CC02 EE13 4C088 AB57 AC05 AC06 AC11 BA08 BA09 BA10 CA05 CA06 CA07 CA09 CA13 NA14 ZA70 ZA89 ZC20 4C206 AA01 AA02 AA04 CB18 MA01 MA04 NA14 ZA70 ZA89 ZC89

Claims (5)

[Claims] 1. A lipase inhibitor comprising as an active ingredient a solvent extract of one or more plant species selected from the genus Eucalyptus. 2. The solvent used for extraction is a lower alcohol, a glycol, a halogenated hydrocarbon, an ether,
2. One solvent selected from lower fatty acid esters, ketones and water, or an arbitrary mixed solvent using two or more of these solvents that can be uniformly mixed. Lipase inhibitor. 3. A plant species selected from the genus Eucalyptus is E. g.
lobulus , E. viminalis , E. camaldulensis , E. grandi
s, lipase inhibitor of claim 1 or claim 2 characterized in that it is a plant species selected from the group consisting of E. nitens. 4. The lipase inhibitor according to any one of claims 1 to 3, wherein the extract contains dihydrochalcone represented by the following chemical formula (1) as a main active ingredient. [Chemical 1] 5. A lipase inhibitor containing dihydrochalcone represented by the following chemical formula (1) as an active ingredient. [Chemical 2]