JP5488870B2 - Olive fruit extract, human leukocyte elastase inhibitor containing the extract, and method for producing olive fruit extract - Google Patents

Description

  The present invention relates to an olive fruit extract, a human leukocyte elastase inhibitor comprising the extract, and a method for producing an olive fruit extract.

  Olive is a plant belonging to the family Moleaceae that originates in the Mediterranean coast, and has been cultivated for food and oil extraction since ancient times. Olive fruit usually grows from around October and becomes pale green, and when it comes to purple-red spots, it is a suitable time for processing into green pickles. After that, around November, the whole becomes a so-called live olives with a purple-black color, which is suitable for salting processing. The ripe olive fruit around December contains about 15-30% oil, and olive oil is extracted from the olive fruit at this time. The olive fruit and olive oil thus obtained are known to have various excellent effects such as arteriosclerosis, gastric ulcer and constipation reduction, bone strengthening, prevention of aging, and skin beautification.

  Above all, olive oil is a vegetable oil containing various active ingredients such as oleic acid, provitamin A, vitamin B, vitamin Ks, polyphenols, etc., and has recently attracted attention for its health improvement effect. Has been reported to have a cancer-inhibiting effect based on phenotypes and polyphenols, and to promote bone formation based on provitamin A, vitamin D and vitamin K. Olive oil also has skin beautifying effects such as reducing rough skin, keeping the skin healthy, moisturizing the skin, protecting the skin, and preventing the skin from drying, and is formulated into cosmetics and topical medicines. ing.

  Moreover, as an olive fruit extract obtained from olive fruit, what is obtained by extracting the fruit and / or seed of an olive plant with water and / or an organic solvent is known (for example, refer patent document 1). This extract has been reported to have an action to remove active oxygen, an inhibitory effect on melanin production, and an inhibitory / killing effect on tumor cell growth, and the solvent used for the extraction provides safety to the human body of the resulting extract. However, from the viewpoint of increasing its action, it is desirable to use a mixed solvent of water and alcohol.

JP 2001-181197 A

As described above, olive fruits and extracts of the fruits contain various active ingredients that have a beneficial effect on the living body. However, these actions are not always sufficient.
Accordingly, an object of the present invention is to provide an olive fruit extract having high physiological activity, a human leukocyte elastase inhibitor comprising the extract, and a method for producing an olive fruit extract.

As a result of diligent research, the inventors of the present invention are an olive fruit extract containing an olive fruit extract extracted from olive fruit cake crushed after freezing, 1,3-butylene glycol, and water. It was found that an olive fruit extract containing the above polyphenol exhibits excellent human leukocyte elastase inhibitory action. In addition, olive fruit extract is crushed after freezing as olive fruit cake, olive fruit is squeezed, then the obtained fruit cake is frozen at a temperature of −10 ° C. or lower, and the obtained frozen rice cake is The present invention was also completed by finding that it exhibits an excellent inhibitory effect on human leukocyte elastase when a product obtained by grinding until the maximum length of the ground piece becomes 1 mm or less is used.

That is, the present invention
(1) An olive fruit extract comprising an olive fruit extract extracted from olive fruit cake ground after freezing, 1,3-butylene glycol, and water,
The olive fruit extract contains, as polyphenols, tyrosol and hydroxytyrosol, an olive fruit extract,
(2) The olive fruit extract also contains other polyphenols, and based on the total mass of polyphenols, the content of tyrosol is 1.5 to 3.5% by mass, and the content of hydroxytyrosol The olive fruit extract according to the above (1), wherein the amount is 0.1 to 1.0% by mass, and the content of oleuropein is less than 0.1% by mass,
(3) The olive fruit cake crushed after freezing is obtained by squeezing the olive fruit, then freezing the obtained fruit cake at a temperature of −10 ° C. or less, and using the obtained frozen rice cake as a maximum of the crushed pieces. The olive fruit extract according to (1) or (2), wherein the olive fruit extract is produced by pulverizing until the length is 1 mm or less,
(4) The olive fruit extract according to (3), wherein the olive fruit variety is Mission (Misson),
(5) a human leukocyte elastase inhibitor comprising the olive fruit extract according to any one of (1) to (4) above,
(6) A method for producing an olive fruit extract, comprising a step of extracting olive fruit cake crushed after freezing with a mixture of 1,3-butylene glycol and water,
It is about.

The present invention will be described in more detail.
The olive fruit extract of the present invention comprises an olive fruit extract extracted from olive fruit cake ground after freezing, 1,3-butylene glycol, and water, and the content of tyrosol is a polyphenol in the extract. 1.5 to 3.5% by mass, based on the total mass of the above, the content of hydroxytyrosol is 0.1 to 1.0% by mass based on the total mass of polyphenols in the extract, The amount is less than 0.1% by weight, based on the total weight of polyphenols in the extract, but the olive fruit cake crushed after freezing is squeezed olive fruit, followed by It is made by freezing the resulting fruit koji and grinding the resulting frozen koji.
Examples of the olive fruit to be used include Mission, Manzanillo, Nevadillo Blanco, Picual, Ojiblanca, Arbequina, C, and Colnica. Gordal, Frantoio, Moroiolo, Recino, Coratina, Ascolana Terena, Olivel (Ol)
ivie), FS17 (Frantoistock 17), Lucca and the like.
A preferable olive fruit variety is, for example, Mission.
Freezing is preferably performed at a temperature of −10 ° C. or lower.
It is preferable to grind the frozen rice cake until the maximum length of the crushed pieces is 1 mm or less.
In addition, for the pulverization of the frozen koji, a normal pulverizer used in the food field can be used, but those that are not heated during pulverization are preferable, and a pulverizer that can be pulverized while cooling is used. preferable.

In the extraction operation, first, the olive fruit cake crushed after freezing is charged into an extraction container with a mixture of 1,3-butylene glycol as an extraction solvent and water.
The ground olive fruit cake can be used for the extraction operation in either a frozen state or a thawed state.

  As the extraction solvent, a 100% 1,3-butylene glycol solvent containing no other auxiliary solvent can be used. However, the extraction solvent may be a mixed solvent composed of water and 1,3-butylene glycol. When a mixed solvent is employed, the concentration is, for example, 30% by volume.

  Extraction is performed after charging the olive fruit koji and the extraction solvent into the extraction container. Extraction can be performed at a temperature slightly higher than room temperature in order to promote the extraction action. For example, when the extraction temperature is 50 to 60 ° C., the extraction time is about 5 hours.

  After the extraction is completed, the olive fruit cake and the extract are separated. Separation of olive fruit can be performed by filtration. If the obtained filtrate is convenient for the subsequent solid content separation, the total amount may be diluted several times.

  Next, filtration is performed again, and the solid content contained in the extract is further separated. Prior to solid content separation, it is preferable to adsorb and separate fine solids by adding an adsorbent such as diatomaceous earth to the liquid to be filtered. The amount of adsorbent added is not particularly limited, but is, for example, 10% by volume with respect to the liquid to be filtered.

  After separating the solid content in this way, the obtained filtrate is sterilized. Sterilization can be performed by ordinary heat sterilization. For example, pasteurization that is maintained at 80 to 90 ° C. for about 1 hour is preferable. High-temperature sterilization using a temperature exceeding 100 ° C. can shorten the sterilization time, but is not a preferable method because it causes decomposition of active ingredients contained in olive fruit straw.

By filtering the sterilized liquid again, an olive extract is obtained. This final filtration is preferably performed using a filter having a fine 0.5 μm cut-off, for example. By such filtration, the amount of solids contained in the olive extract can be reduced to 0.5% or less.
The olive extract contains tyrosol and hydroxytyrosol as polyphenols.
The olive extract also contains other polyphenols, and based on the total mass of the polyphenols, the content of tyrosol is 1.5 to 3.5% by mass, and the content of hydroxytyrosol is 0.1 to 1.0% by mass, and the content of oleuropein is less than 0.1% by mass.
The olive extract of the present invention is produced by diluting the olive extract obtained above as it is or as necessary with 1,3-butylene glycol and water.

  The olive fruit extract of the present invention thus produced has an excellent human leukocyte elastase inhibitory action. These actions will be briefly described below.

-Human leukocyte elastase inhibitory action Elastin is an elastic tissue contained in the skin, and when it is decomposed, the skin gradually loses its tension and elasticity. Elastin is degraded by elastase, and elastase is activated by aging and by external and internal stimuli such as ultraviolet rays. Therefore, if the activity of elastase, which is a degrading enzyme of elastin, can be inhibited to suppress the degradation of elastin, the elasticity of the skin can be maintained and the skin can be kept firm and moist.
In addition, diseases caused by elastin degradation by elastase, collagen fiber degradation and / or abnormally increased proteoglycan degradation in humans, such as chronic obstructive pulmonary diseases such as emphysema, rheumatoid arthritis, atherosclerosis, adult respiratory distress It is also useful for the treatment and / or prevention of (ARDS), glomerulonephritis, myocardial infarction, ulcerative colitis, periodontal disease and the like.

  The olive fruit extract of the present invention can be in various dosage forms. For example, it can be blended in foods, beverages, pharmaceuticals and the like taken orally. Above all, since it exhibits an excellent skin beautifying effect, it is preferably blended with external preparations applied directly to the skin, such as ointments, creams, emulsions, suspensions, solutions and the like. The compounding quantity to these is 1 to 50 weight%, for example.

  The following examples illustrate the invention in more detail, but these examples are not intended to limit the invention to certain specific embodiments.

Examples 1 to 3: Production of olive fruit extract Olive fruit extract was produced as follows.
Preparation: Olive fruit 100 g after squeezing olive fruit of variety mission (Misson) was frozen in a freezer for food (−15 ° C.). The obtained olive frozen fruit cake was pulverized with a food pulverizer until the maximum length of the pulverized pieces became 1 mm or less. The obtained pulverized product and 1000 g of a 30% 1,3-butylene glycol aqueous solution were charged into an extraction container.
Extraction: The extraction vessel was heated to keep the interior at 50-60 ° C. Extraction was carried out with stirring at this temperature for 5 hours.
Olive fruit koji separation: After the completion of extraction, the olive fruit koji was removed and 1000 g of 30% 1,3-butylene glycol aqueous solution was added to dilute the extract twice.
Solid content separation: 200 g of diatomaceous earth (Radiolite # 200) was added to the diluted solution and stirred for 1 hour to adsorb fine solid matter. Thereafter, the diluted solution was filtered to separate solids and diatomaceous earth.
Sterilization: The filtrate was heated to 80 to 90 ° C. and sterilized by heating with stirring for 1 hour.
Final filtration: The sterilized filtrate was final filtered using a 0.45 μm cut-off filter.
According to the above procedure, 1541.2 g of olive fruit extract containing 0.38% solid content was obtained.
The same operation as described above was performed to produce a total of 3 lots of olive fruit extract.
A table summarizing the quantification results of the three lots produced is shown below.
The amount of polyphenol (total amount) was measured by the folin-thiocult method, and the amounts of oleuropein, hydroxytyrosol and tyrosol were determined by HPLC.

Test Example 1: Evaluation of Elastase Activity Inhibitory Action Except for using the 30% 1,3-butylene glycol aqueous solution instead of the olive fruit extract obtained in Example 1, the same process as in Example 1 was used. The elastase activity inhibitory action of the olive fruit water extract obtained in 1 was evaluated. The reason for using water for extraction is that elastase activity cannot be measured when the sample contains alcohol.
50 μL of olive fruit water extract, 25 μL of 12 μg / mL human leukocyte-derived elastase (ELASTIN PRODUCT CO. INC), and 25 μL of methoxy-succinyl-alanyl-alanyl-propylvaline-p-nitroanilide prepared to 8 mM, The reaction was carried out at 37 ° C. for 20 minutes. The solution after the reaction was measured for absorbance at 405 nm, and the produced nitroaniline was quantified.
As a result, 37.9% of human leukocyte elastase activity was inhibited.
The results obtained for the water extract are themselves good. However, it is known in the prior art that substances having elastase activity inhibitory ability are polysaccharides such as peptin, and polysaccharides are generally more efficient when using water and 1,3-butylene glycol than ethanol. It is also known that it can be extracted. Therefore, the olive fruit extract of Example 1 is expected to exhibit elastase activity that is much higher than that of the water extract tested in this test example, and much higher than that of the ethanol extract, which is much superior to that of the ethanol extract.

表２から解るように、本発明のオリーブ果実エキスを配合したハンドクリームは、比較のハンドクリームを越える著しい美肌作用を奏した。 Test Example 2: Production of Olive Cream Extract-Containing Hand Cream In addition to the following basic components, 3% by weight of the olive fruit extract of Example 1 was blended with respect to the total of other components to obtain a hand cream.
Basic ingredients: water, mineral oil, petrolatum, glycerin, hydrogenated polyisobutene, microcrystalline wax, cyclomethicone, lanolin alcohol, paraffin, squalane, jojoba oil, decyl oleate, octyldodecanol, aluminum distearate, magnesium stearate, sulfuric acid Magnesium, citric acid, benzoate, tocopherol, butylhydroxyanisole, dibutylhydroxytoluene, fragrance.
As a comparison, an olive fruit extract was obtained by performing the same operation as in Example 1 except that 100 g of frozen olive fruit was used instead of the pulverized product obtained by freezing 100 g of olive fruit cake after oil extraction. A hand cream having the same composition as described above was produced.
The hand cream thus produced and the comparative hand cream were applied to the subject's skin once a day, and the change in the applied part after 10 days was moistened, and the three items of tension and gloss were observed.
The results are shown in Table 2. The results in Table 2 show the number of subjects improved for each item.

As can be seen from Table 2, the hand cream formulated with the olive fruit extract of the present invention exhibited a remarkable skin beautifying action over the comparative hand cream.

表３から解るように、本発明のオリーブ果実エキスを配合したボディーソープは、比較のボディーソープを越える著しい美肌作用を奏した。 Test Example 3 Production of Body Soap Containing Olive Fruit Extract In addition to the following basic components, 2% by weight of the olive fruit extract of Example 1 was blended with respect to the total of other components to obtain a body soap.
Basic ingredients:
Ammonium lauryl sulfate 8%
Sodium polyoxyethylene lauryl ether sulfate 2%
Palm oil fatty acid amidopropyl betaine 7%
Palm oil fatty acid monoethanolamide 4%
Edetic acid disodium 0.2%
Sodium benzoate 0.2%
Citric acid 0.2%
The remainder The body soap produced in this way and the comparative body soap having the same composition using the comparative olive fruit extract obtained in the same manner as in Test Example 2 were used by the subject once a day, and the skin changes after 10 days. The three items of moisture and tension and gloss were observed.
The results are shown in Table 3. The results in Table 3 show the number of subjects improved for each item.

As can be seen from Table 3, the body soap formulated with the olive fruit extract of the present invention exhibited a remarkable skin beautifying action over the comparative body soap.

表４から解るように、本発明のオリーブ果実エキスを配合した美容液は、比較の美容液を越える著しい美肌作用を奏した。 Test Example 4: Production (prescription) of cosmetic liquid containing olive fruit extract
(Phase A)
Ethyl alcohol (95%) 10.0% by mass
Polyoxyethylene (20 mol) octyldodecanol 1.0
Pantotenyl ethyl ether 0.1
Olive fruit extract of the present invention 3.0
Methylparaben 0.15
(Phase B)
Potassium hydroxide 0.1
(Phase C)
Glycerin 5.0
Dipropylene glycol 10.0
Sodium bisulfite 0.03
Carboxyvinyl polymer 0.2
Purified water residue (production method)
A phase and a C phase were uniformly dissolved, the A phase was added to the C phase to solubilize, and then a B phase was added to obtain a cosmetic liquid.
The cosmetic liquid thus prepared and the comparative cosmetic liquid having the same composition using the comparative olive fruit extract obtained in the same manner as in Test Example 2 were applied to the skin of the subject once a day, The change was moistened, and the three items of tension and gloss were observed.
The results are shown in Table 4. The results in Table 4 show the number of subjects improved for each item.

As can be seen from Table 4, the essence containing the olive fruit extract of the present invention exhibited a significant skin beautifying effect that exceeded that of the comparative essence.

表５から解るように、本発明のオリーブ果実エキスを配合したシャンプーは、比較のシャンプーを越える優れた効果を示した。 Test Example 5: Production of shampoo containing olive fruit extract-60 g sodium lauryl ether sulfate (28% AM)
-9g cocoyl betaine
-2 g of olive fruit extract of the present invention
-Preservative, perfume Appropriate amount-Amount to make HCl pH 6-Amount to make water 100 g The milky white shampoo was obtained by blending the above ingredients.
The shampoo produced in this manner and the comparative shampoo having the same composition using the comparative olive fruit extract obtained in the same manner as in Test Example 2 were used to wash the hair using the subject once a day, and the scalp and hair after 10 days. The overall change was moistened, and three items of tension and gloss were observed.
The results are shown in Table 5. The results in Table 5 show the number of subjects improved for each item.

As can be seen from Table 5, the shampoo containing the olive fruit extract of the present invention showed an excellent effect over the comparative shampoo.

  According to the present invention, the olive fruit cake after oil extraction that has been crushed after freezing is extracted with 1,3-dibutyl ether-water, so that the elastase activity inhibitory action is significantly superior to conventional products. A fruit extract can be provided. Such an olive fruit extract can be preferably blended with external medicines such as ointments, creams, emulsions, suspensions and solutions.

Claims (4)

An olive fruit extract containing an olive fruit extract extracted from olive fruit cake crushed after freezing, 1,3-butylene glycol, and water,
The olive fruit cake crushed after freezing is obtained by squeezing olive fruit, subsequently freezing the obtained fruit cake at a temperature of −10 ° C. or less, and obtaining the obtained frozen rice cake with the maximum length of the crushed pieces. Is made by crushing until it becomes 1 mm or less,
The olive fruit extract contains, as polyphenols, tyrosol , hydroxytyrosol and other polyphenols, and the content of tyrosol is 1.5 to 3.5% by mass, respectively, based on the total mass of polyphenols, An olive fruit extract characterized in that the content of hydroxytyrosol is 0.1 to 1.0% by mass and the content of oleuropein is less than 0.1% by mass . The olive fruit extract according to claim 1 , wherein the olive fruit variety is Mission. A human leukocyte elastase inhibitor comprising the olive fruit extract according to claim 1 or 2 . The method for producing an olive fruit extract according to claim 1 or 2, comprising a step of extracting the olive fruit cake crushed after freezing with a mixed solution of 1,3-butylene glycol and water.