JP2006298812A - Antioxidant, skin lotion for anti-oxidization, elastase activity inhibitor and maillard reaction inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To utilize an extract of papaya (Carica papaya L.), on which only an effect as a health food is found out until now, effectively. <P>SOLUTION: This papaya extract is found to exhibit a potent esterase activity inhibitory effect, Maillard reaction inhibitory effect and also SOD (superoxide dismutase) like activity, prevent various diseases such as senile phenomena accompanying with aging, diabetes, arteriosclelosis, etc., and also improve various skin troubles such as skin roughness, sensitive skin, dermatitis diseases, etc. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

The product of the present invention is characterized by the fact that an extract extracted from a plant of a specific genus significantly suppresses elastase activity and Maillard reaction, and further exerts an SOD-like activity action, thereby causing aging with age, diabetes, arteriosclerosis The present invention relates to antioxidants, antioxidant skin external preparations, elastase activity inhibitors, Maillard reaction inhibitors applied to pharmaceuticals, quasi-drugs, foods and the like that prevent various diseases such as the above.

Papaya is a fruit native to tropical America that is popular for its ripe, soft fruit and moderate sweetness. Until now, papaya has been known to have an effect of promoting keratinization, and a food for preventing fine wrinkles containing aloe having an excellent antioxidant function has been disclosed (see Patent Document 1). It is also disclosed as a cosmetic composition (see Patent Documents 2 to 5). However, its use as an antioxidant is not known.

There are various causes of skin aging, and there is a decrease in collagen and hyaluronic acid. One of them is elastin that maintains the physical structure of the skin. Degeneration and destruction of elastin due to excess of elastin, an elastin-destroying enzyme due to sunlight (ultraviolet rays), drying, etc., leads to a decrease in skin elasticity, causing wrinkles and tarmi. .
Therefore, it is clear that suppressing the action of elastase and preventing the degeneration and destruction of elastin which gives elasticity and elasticity to the skin suppresses wrinkles and tarmi, and thus contributes to the prevention of skin aging.

In addition, when elastase is excessive, elastin which forms the main component of the elastic fiber is decomposed to cause tissue damage, various inflammations or degenerative conditions. Such excessive degradation of elastin by elastase is considered to be the cause of diseases such as emphysema, adult respiratory distress syndrome, pulmonary fibrosis, bronchitis, pneumonia, rheumatoid arthritis, arteriosclerosis, sepsis, shock, pancreatitis, nephritis . Accordingly, elastase inhibitors are considered useful as therapeutic and / or prophylactic agents for these diseases.
A typical example of the Maillard reaction is that a protein reacts with an amino acid and is colored by heating of food or the like, but it is known that this reaction occurs in vivo. When the Maillard reaction occurs in the living body, the function of the protein is impaired, and the function of the collagen is reduced. In recent studies, attention has been focused on the Maillard reaction as a cause of various diseases associated with diabetes, arteriosclerosis and aging associated with aging.

In recent years, there are concerns about the effects of active oxygen, which has a strong oxidizing power, on the living body, but the skin is not only exposed to oxygen stress caused by the body, but is also in contact with air and directly exposed to ultraviolet rays. Therefore, it can be said that the influence of active oxygen is far greater than other organs. This oxidative stress causes various skin troubles such as skin irritation and hypersensitivity, and skin inflammation diseases. Therefore, suppressing the production of active oxygen is an indispensable element for cosmetics.

Several mechanisms have been elucidated for the elimination of active oxygen in the living body. The first O · ^2- (superoxide) is enzymatically converted into H ² O ² and O ² by SOD (superoxide dismutase). It is known to be disproportionated. However, the amount of SOD decreases with aging, and the decrease in the amount of SOD increases the superoxide concentration, leading to injury. Therefore, a substance having the same action as SOD is considered to be effective as an antioxidant in the skin.
JP 11-253129 A Japanese Patent Laid-Open No. 2003-081751 JP 2003-155213 A Japanese Patent Laid-Open No. 2001-039823 JP 2001-226219 A

The object of the present invention has been variously studied in order to effectively utilize an extract of highly effective papaya (Carica papaya L).

In order to achieve the above object, the present inventors have intensively studied and found that the papaya extract has a strong elastase activity inhibitory effect and a Maillard reaction inhibitory effect, and further exhibits a SOD-like activity. It has been found that various diseases such as aging with age, diabetes and arteriosclerosis are prevented.

The form of the papaya extract is not limited in any way. Preferably, the papaya is shade-dried, pulverized and then immersed in an extraction solvent (for example, alcohol such as ethanol, water, or a mixture thereof) for 3 days or 100 degrees for 2 hours. Use the solution heated, cooled, and filtered in.

Test Example 1 Elastase Activity Inhibition Test After culturing fibroblasts in a flask or petri dish (100 mmφ) until it becomes 80-90% confluent, it is replaced with a serum-free medium and IL-1 is adjusted to 0.4 ng / ml. And incubate for 48 hours. The medium was discarded, the cells were detached with 0.1% Triton-X100 / 0.1 M Tris-HCl buffer pH 8.3, sonicated under ice-cooling, and centrifuged to obtain an enzyme solution.
The test was performed with a sample (Sample), background (BG), control (Cont.), Blank (BL) and positive control (EDTA).
Place 178 μL of enzyme solution and 20 μL of sample in a 500 μL Eppendorf tube (BG and BL add 0.1% Triton-X100 / 0.1M Tris-HCl buffer pH 8.3 instead of enzyme solution. Cont. And EDTA are samples of (Add water or 100 mmol / L EDTA-2Na aqueous solution instead) Let react for hours. After adding 100 μL of acetic acid and stopping the reaction, the absorbance at 405 nm of the supernatant centrifuged with a desktop centrifuge was measured with a microplate reader, and the inhibition rate was calculated by the following formula.
Inhibition rate = (((Cont.−BL) − (Sample−BG)) / (Cont.−BL)) × 100
The test products are shown below, and the test results are shown in Table 1.
Papaya E: Papaya (fruit dried product) (20 g) ethanol (100 ml) extract (lyophilized product)
Papaya EW: Papaya (fruit dried product) (20 g) 50% ethanol-water (100 ml) extract (lyophilized product)
Papaya W: Papaya (dried fruit) (20 g) hot water (100 ml) extract (freeze dried)

Test Example 2 Maillard Reaction Inhibition Test Maillard reaction inhibitory action was examined on the extract of papaya (Carica papaya L.) by the NBT reduction method that measures ketoamine, which is an intermediate of Maillard reaction. 100 μl of purified water and 500 μl of 0.1M phosphate buffer (pH 6.0) were added to 50 μl of a test product prepared with water, and 100 μl of 2M glucose aqueous solution was added. Furthermore, 200 μl of 4 mg / ml BSA (Bovine Serum Albumin) aqueous solution was added, stirred, and incubated at 60 ° C. for 30 hours. After 30 hours, 2.7 ml of 0.2 mg / 0.9 ml [phosphate buffer (pH 10.3)] NBT (nitroblue tetrazolium chloride) solution was added to 300 μl of the reaction solution, and incubated at 37 ° C. for 30 minutes. Absorbance (A) was measured at 530 nm. Similarly, absorbance (B) when water is used as an alternative to an aqueous BSA solution, absorbance (C) when water is used as an alternative to a test product, water as an alternative to an aqueous BSA solution, and water as an alternative to a test product Absorbance (D) when used was measured, and Maillard reaction inhibition rate (%) was calculated from the following formula.
<Formula> Maillard reaction inhibition rate (%) = [1- (A−B) / (C−D)] × 100
The test products are shown below, and the test results are shown in Table 2.
Papaya E: Papaya (fruit dried product) (20 g) ethanol (100 ml) extract (lyophilized product)
Papaya EW: Papaya (fruit dried product) (20 g) 50% ethanol-water (100 ml) extract (lyophilized product)
Papaya W: Papaya (dried fruit) (20 g) hot water (100 ml) extract (freeze dried)

Test 3: SOD-like activity test
Specimen solution (H ₂ for the subject) was added to 0.6 ml of test solution in which equal amounts of KH ₂ PO ₄ -borax buffer solution, 0.5 mM xanthine solution and 0.25 mM hydroxyamine hydrochloride solution containing 0.5 mM EDTA-2Na were mixed. O) 0.2 ml and 1.43 μL / ml xanthine oxidase solution (Wako Pure Chemical Industries milk xanthine oxidase suspension 5.9 unit / ml) 0.2 ml were added and incubated at 37 ° C. for 30 minutes. Thereafter, 2 ml of N-1 naphthylethylenediamine-sulfanilic acid acetic acid solution was added to stop the reaction, left at room temperature for 30 minutes, and the absorbance at 550 nm was measured with an absorptiometer. The erasure rate was calculated by the following method.
Erase rate = (A−B) / A × 100
A: Absorbance difference when adding air solution and enzyme solution in the absence of sample
B: The difference in absorbance when the air solution and the enzyme solution were added in the presence of the sample and the sample are shown below, and the test results are shown in Table 1.
Papaya E: Papaya (30 g) ethanol (200 ml) extract (lyophilized)
Papaya EW: 50% ethanol-water (200 ml) extract of papaya (30 g) (lyophilized product)
Papaya W: Papaya (30 g) hot water (200 ml) extract (lyophilized)
I went there.

Antioxidants, antioxidant skin external preparations, elastase activity inhibitors, Maillard reaction inhibitors of the present invention are used as external preparations (pharmaceuticals, quasi-drugs, cosmetics) as internal preparations (pharmaceuticals, foods) according to conventional methods. It can be prepared by blending with various known forms of substrates.
It does not specifically limit as a form of an external preparation, For example, arbitrary dosage forms, such as a milky lotion, cream, aqueous solution, a pack, can be selected.
Moreover, as an internal preparation, the form as a tablet, a granule, a capsule, a drink, or a foodstuff may be sufficient.

In the external preparation or internal preparation of the present invention, in addition to the above-mentioned essential components, components to be blended in a normal external preparation or internal preparation, for example, oil agent, powder, purified water, polymer compound, gelling agent , UV absorbers, UV scattering agents, antioxidants, pigments, preservatives, fragrances, and cosmetic ingredients can be appropriately selected and used within a range not impairing the effects of the present invention.

  EXAMPLES Next, although an Example and a comparative example are given and this invention is demonstrated still in detail, this invention is not restrict | limited at all to these. Moreover, the extraction method about the extract of the used chemical | medical agent is not limited at all.

Tables 4 to 8 show the formulations of Examples and Comparative Examples. The preparation method was performed by a conventional method. Table 4 is a tablet, Table 5 is a beverage, Table 6 is a lotion, Table 7 is a cosmetic solution, Table 8 is a cream formulation, and the amount is shown in parts by weight.

Details of the raw materials used in the above comparative examples and examples are described below.
(Note 1) Papaya extract 1 was prepared by adding 200 ml of purified water to 10 g of papaya (dried fruit) and boiled for 1 hour, and then using the filtered extract.
(Note 2) Papaya extract 2 was prepared by adding 100 ml of purified water and 100 ml of ethanol to 10 g of papaya (dried fruit) and stirring for 5 days, and then using the filtered extract.
(Note 3) Papaya extract 3 was prepared by adding 200 ml of ethanol to 10 g of papaya (dried fruit) and stirring for 5 days, and then using the filtered extract.
(Note 4) The hydrolyzed conchiolin solution used was a hydrolyzed conchyolin that had a molecular weight of 1000 or less.
(Note 5) As the tablinus extract, a Matils extract [manufactured by Maruzen Pharmaceutical Co., Ltd.] was used.
(Note 6) Falco Rex Honoki E [Ichimaru Falcos Co., Ltd.] was used as the honoki extract.
(Note 7) As the guava extract, 300 ml of ethanol was added to 10 g of guava leaves and allowed to stand for 5 days, and then the filtered extract was used.
(Note 8) Nishi-Haruyanagi extract used Nishi-Haruyanagi extract [Nippon Seika Co., Ltd.].
(Note 9) Marine purge [manufactured by Ichimaru Falcos Co., Ltd.] was used as the seaweed extract.
(Note 10) As the acacia bark extract, 300 ml of ethanol was added to 10 g of bark and left for 5 days, and then the filtered extract was used.
(Note 11) The Murayako Engii extract used was the Murayako Engii extract [Yamakawa Trading Co., Ltd.].
(Note 12) As for the extract of the giant crape myrtle, the extract of the giant crape myrtle (produced by Yamakawa Trading Co., Ltd.) was used.

Confirmation of effect 1
The effect test of Examples 1-2 of Table 4-Table 5 and Comparative Examples 1-2 was implemented. The test method consisted of 20 females aged 25 to 55 years old, taking a proper amount of the test food after meals twice a day in the morning and at night for 12 weeks. Table 9 shows the results of the effect of improving skin troubles by taking.

  Based on the above results, papaya extract blended with foods and drinks improved various skin problems such as wrinkles, spots / freckle, dullness, and rough skin, and decreased blood glucose levels among testers. In addition, many subjects complained of improved health.

Confirmation of effect 2
Examples 3 to 14 described in Tables 6 to 8 Effect tests of Comparative Examples 3 to 5 were performed. The test method consisted of 20 females aged 25 to 55 years old, and a suitable amount of a topical preparation was applied to the face after washing the face twice a day in the morning and night for 12 weeks. Table 10 shows the results of the skin trouble improving effect by application.

Claims (4)

Antioxidant characterized by blending papaya (Carica papaya L.) extract Antioxidant skin external preparation characterized by containing papaya (Carica papaya L.) extract Elastase activity inhibitor characterized by containing papaya (Carica papaya L.) extract Maillard reaction inhibitor characterized by containing papaya (Carica papaya L.) extract