JP3533737B2 - Bioaging agent and skin composition - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a biological anti-aging agent and a dermatological composition which have a high activating effect on living cells and tissues such as skin tissue and have an excellent anti-aging effect.

[0002]

2. Description of the Related Art The mechanism of aging has been actively studied,
Various theories are presented. However, it is still in the basic research stage, and it is hard to say that effective measures to prevent aging, such as prolonging lifespan and delaying various symptoms characteristic of aging, have been established. In such a situation, although few in number, some attempts have been made at the individual level and the organizational level.

Tissue level studies include attempts at aging the skin and joints. In the skin and joints, when cellular function declines due to aging, not only the peroxidation of biological components but also the amount of hyaluronic acid in the tissues decreases, resulting in dry skin, loss of firmness / elasticity, and wrinkles / wrinkles. Increase, decrease in lubricity of joints. On the other hand, two typical countermeasures are known. One is a method in which a cosmetic containing hyaluronic acid, a moisturizing component, etc. is applied to the skin, or hyaluronic acid is directly injected into the joint. The other is a method of applying retinoic acid, which promotes hyaluronic acid synthesis in the living body and leads to improvement of wrinkles. However, the former does not lead to a fundamental improvement, and the latter retinoic acid has strong side effects such as strong skin irritation and skin thickening.

On the other hand, in the individual-level study, it was discovered from the results of the animal experiment that the animals were fed with a feed lacking a specific nutrient, vitamin E, vitamin C, etc. have the effect of delaying the aging phenomenon. For example, vitamin E has the strongest effect. Therefore, various studies have been carried out with the expectation that vitamin E has a life extension effect on mice and rats. However, so far, vitamin E has not been found to have a clear life-prolonging effect.

However, subsequent studies have revealed that even with the effects of vitamin E and vitamin C levels, which do not show a clear life-prolonging effect, local administration thereof suppresses lipid peroxidation due to skin spots, wrinkles, and aging. It is widely used as an anti-aging agent not only in pharmaceuticals but also in cosmetics and foods. At present, it is considered that these substances act as antioxidants and suppress the oxidative damage caused by active oxygen generated by oxygen metabolism, and as a result, exhibit anti-aging action.

[0006] As described above, various studies and proposals have been made on the active ingredient which exerts anti-aging effect, but an active ingredient having excellent anti-aging effect and high safety is desired.

[0007]

Means and Actions for Solving the Problems The present inventors have
As a result of extensive studies to meet the above demand, Coleus forskohli
The root extract of i) or forskolin is a nematode C. It has a long-lasting effect on elegans and a high hyaluronan synthesis-stimulating effect on rat epidermal cells, and these effects are more significantly enhanced when used in combination with tocotrienols, and thus Coleus forskolini Root extract or forskolin and tocotri
It has been found that the combination with enol has an excellent activating effect on living cell tissues such as skin tissues and has a high anti-aging effect.

That is, C.I. Elegance is widely known as a model animal suitable for aging research. The aging process has been studied at the genetic level, and it is an essential model system for aging research in mammals including humans (J
individual of theAmerican Ger
iatrics Society 40 (9), 199
2, 936-945). The average lifespan is genetically controlled, is constant (approximately 20 days at 20 ° C), and the metabolic activity of cells and tissues decreases with age, and the process similar to that of higher animals such as the accumulation of peroxides. It leads to death. Therefore, C.I. Elegance life span is a good indicator of cell and tissue aging. Therefore, the nematode C. C. When the life of elegans can be extended, it can be evaluated that this component has an activating effect on living cells and tissues including skin and also has an anti-aging effect.

On the other hand, hyaluronic acid synthesizing ability catches the skin aging phenomenon. With age, the skin loses firmness and luster, and wrinkles increase. It is a well-known fact that this is due to a decrease in elasticity and water retention of the skin, particularly the epidermis. Hyaluronic acid retains a large amount of water in the skin and greatly contributes to maintaining homeostasis such as elasticity and water retention of the skin. It is known that this hyaluronic acid decreases with aging, which leads to skin firmness, loss of gloss, and increase of wrinkles. That is, increasing hyaluronic acid in the epidermis is nothing more than suppressing aging of the skin, and one component having a hyaluronic acid synthesis promoting effect means that this component suppresses aging of skin cells and tissues. It can be evaluated as having an activation effect.

From the above points, the present inventors have found that C. elegans C. The effect of prolonging the lifespan of elegans and the effect of promoting hyaluronan synthesis on rat epidermal cells were selected as screening indices, and as a result of repeated screening,
Forskolin root extract or forskolin and toco
They have found that the desired effect can be obtained in combination with trienol, and have completed the present invention.

It has been known that an extract of roots of Choleus forskolini or forskolin has been used as an active ingredient of an external preparation for skin or a composition for hair (JP-A-63-304978 and JP-A-64978). -79107
No. 1,216,912, and No. 3-188024.
No., Publication No. Hei 5-501108) is an extract of roots of Coleus, false Koryi or forskolin and preparative
It is a new finding by the present inventors that the combined use with cotrienol gives a high anti-aging effect.

Therefore, the present invention provides a root extract of Coleus forskolini or forskolin and tocotrienos.
The present invention provides a biological anti-aging agent containing glutinol as an active ingredient, and a skin composition containing the biological anti-aging agent.

The present invention will be described in more detail below.

The Coleus forskohlii used in the present invention belongs to the Labiatae family and has been used as a folk medicine for a long time in regions such as India, Arab, Africa and Brazil, and has a heart and respiratory system. It is also said to be effective against diseases of the nervous system or abdominal pain. In India, this is eaten as an accessory. As Choleus forskolii used in the present invention, those cultivated and grown naturally in these areas can be used, but are not limited thereto. For example, Coleus forskolii cultivated in Japan is no different from the one obtained from the above region in terms of composition and effectiveness, and thus these can be effectively used.
In addition, microtuber, adventitious bud, adventitious root, hairy root,
An organ obtained by a tissue culture method such as callus can also be used.

There is no limitation on the method for obtaining the root extract of Coleus forskoli applied to the present invention, and a usual extraction method is adopted, and a hydrophilic organic solvent such as water, a water-containing hydrophilic organic solvent, and other organic substances are used. Extracted using a solvent or the like, a known root extract of Choleus forskolii can be used in the present invention.

Forskolin is a known compound found in the roots of Coleus forskolini. The following method is suitable as a method for obtaining a root extract containing this substance. That is, it is a method in which dried roots of Coleus forskolini are cold-soaked with methanol or ethanol to obtain an extract, which is concentrated to obtain an extract. Incidentally, this method shows one embodiment as a method for obtaining a root extract of Choleus forskolini containing forskolin of the present invention, in the production process according to the present invention, the solvent used for extraction and its The order of use is not limited and can be arbitrarily selected.

In the present invention, as the root extract of Choleus forskolii, when the extraction solvent is a non-toxic one such as water, ethanol or water-ethanol, the extract may be used as it is, or It can be used as a diluent. Further, it may be a concentrated extract, and may be made into a dry powder product by freeze-drying or the like, or may be prepared as a paste. If necessary, a purification step may be added to increase the content of forskolin in the extract, and the final extract may be forskolin.

[0018] On the other hand, in the present invention, using the door Birds enol
To use.

The ratio of the above-mentioned tocotrienol to be used in combination with the root extract of Choleus forskolini or forskolin is appropriately selected, but is 0.01 to 50 parts by weight, particularly 0.1 to 10 parts by weight to 1 part by weight of forskolin. It is preferable that

The biological anti-aging agent of the present invention is excellent in cells and tissue activating effects on skin tissues and the like, and has a high hyaluronic acid synthesis accelerating ability. Therefore, it is suitable for skin compositions such as skin cosmetics and skin external preparations. In addition to being blended in, it can be blended and used in hair compositions such as hair cosmetics, foods and drinks, and pharmaceuticals. In this case, the dosage form includes liniment, spray, lotion, aqueous solution, emulsion, ointment, powder,
It can be made into external forms such as dissolving tablets and soaps.
Tablets, capsules, powders, oral liquids, fine granules, granules and the like can also be in the form of internal preparations, and the form of the composition in which the bioaging inhibitor of the present invention is blended is not particularly limited. Absent.

When the above-mentioned biological anti-aging agent is compounded in the composition for skin as described above, the compounding amount can be variously selected, but usually 0.001 to 30% by weight of the whole composition, particularly 0.1. It is from 01 to 10% by weight.

As a component constituting a composition for skin or the like to which the bio-aging agent of the present invention is added, a known component may be used depending on the type of the composition, purpose of use, form, etc. The compounding amount can also be a usual dose.

[0023]

As described above, according to the present invention, a biological anti-aging agent according to the present invention, a root extract or forskolin Coleus-false Koryi, by the synergistic action of the bets bird enol, excellent cell, activation of the tissue It has an aging effect, has a high effect of preventing bioaging, and is highly safe. A composition for skin containing this bioaging agent has a high effect of suppressing aging of the skin.

[0024]

EXAMPLES The present invention will be specifically described below by showing Examples and Comparative Examples, but the present invention is not limited to the following Examples.

[Production Example] Production of Choleus forskolii root extract 2 kg of Coleus forskolii root was cold-soaked with 20 L of ethanol three times for 3 days each to obtain an extract. The solvent was distilled off from this extract using an evaporator to obtain 1 L of concentrated extract.

Production of purified extract of Coleus forskolii
The concentrated extract obtained above was subjected to column chromatography [carrier: silica gel (Wakogel C-200), mobile phase: benzene / ethyl acetate = 90/10]. Furthermore, a reversed phase silica gel column [ODS-AQ (250 x
30 mmD. ); YMC Co.] column was used for preparative separation by high performance liquid chromatography. 65% solvent
Acetonitrile, flow rate 40 ml / min, UV 21
Detection was at 0 nm. A portion having a retention time of 11 minutes to 12 minutes was collected and concentrated to dryness to obtain 4.2 g of a light brown powder.

Experimental Example 1 C.I. Evaluation of life extension ability of elegance Escherichia coli st on NG medium
The rain OP50 is cultivated and the NGM medium (nemat
ode growth medium). next,
On this NGM medium, the nematode C. Elegance ( Caeno
rhabditis elegans var. Br
a large amount of istoll) was raised. C. at the same growth stage. To obtain elegance, eggs were collected from a large population of individuals. That is, by immersing a large amount of adults in sodium hydroxide and sodium hypochlorite and melting the body surface of the adults, only eggs could be obtained. The eggs were thoroughly washed in S buffer and allowed to stand at 20 ° C. for 18 hours. The hatched larvae were collected and bred for 3 days on NGM medium until they became adults. 5-Fluoro-2'-deoxyuridine was added to the medium from the 3rd day for the purpose of suppressing hatching emigration of the next-generation larvae. The sample obtained in the above Production Example, retinoic acid, flavin adenine dinucleotide, ascorbic acid, α-tocopherol, coenzyme Q, tocotrienol was dissolved in a very small amount of ethanol and Tween 80, so that the final concentration was 2 mg / ml. Adjusted with water. 4m of this solution so that the final concentration of the sample is 25 μg / ml.
1 of NGM medium was added. From the 4th day of growing up as an adult,
C. Elegance was bred on a sample-added medium and its life span was investigated. The results are shown in Table 1. Table 1 shows the average lifespan obtained from the lifespan curve of the population to which each sample was administered.

[0028]

[Table 1]

Next, flavin adenine dinucleotide,
Ascorbic acid, α-tocopherol, coenzyme Q, and tocotrienol were administered in combination with a purified extract of Coleus forskolini (hereinafter, simply referred to as a purified extract), and the change in life span was investigated by the same method as described above.
12 μg / ml for purified extract, 25 μg / m for other samples
It was set to l. The results are shown in Table 2.

[0030]

[Table 2]

As is clear from the results shown in Tables 1 and 2, the combined use of the refined extract and the antioxidant resulted in a longer life-prolonging effect than that of the single product. Among them, the synergistic effect due to the combination with tocotrienol is particularly remarkable and far surpasses the others. The same result was obtained even when the Coleus forskolii root extract was used.

[Experimental Example 2] Evaluation of Hyaluronic Acid Synthesis Promoting Ability Using Cultured Epidermal Cells Keratinocytes were isolated from the skin of neonatal (3-day-old) rats by 0.25% trypsin treatment, and a growth medium (0.1 mM) was isolated. -Ca) was cultivated until the petri dish was full, and then cultivated in a differentiation medium (1.8 mM-Ca) for 2 days. This cell was used for screening. A sample such as Coleus forskolii root extract is added to the cells at a concentration of 20 μg / ml to 500 μg / ml.
After allowing it to act for ³ hours, ³ H-glucosamine (5 μCi / d
ish) was added and the cells were further cultured for 24 hours. Further, the medium was treated with pronase to digest the protein, and then the total ³ H-GAG containing hyaluronic acid released into the medium was separated by the cetylpyridinium chloride precipitation method, and its radioactivity was measured. GAG when
The ability to promote synthesis (control ratio: magnification) was calculated. The results are shown in Table 3.

[0033] GAG synthesis promoting ability ^{(magnification) = A / B A: 3} H-GAG radioactivity upon addition of sample (dpm) B: the case of no addition ³ H-GAG radioactivity (dpm)

[0034]

[Table 3]

From the results shown in Table 3, it is recognized that the combination of the purified extract and tocotrienol (antioxidant) gives excellent GAG synthesis promoting activity. Also, the same result was obtained when the Coleus forskolii root extract was used. It should be noted that vitamin E has a particularly high G
No AG synthesis promoting activity was observed.

Next, formulation examples of the composition containing the bio-aging agent of the present invention will be shown.

[0037]

[0038]

[0039]

[Formulation Example 4] Cosmetic cream A Oil phase part Liquid paraffin (# 70) 5.0% Squalane 15.0 Cetostearyl alcohol 5.0 Beeswax 2.0 Glycerin monostearate 2.0 POE (20 ) Sorbitan monolaurate 2.0 Propyl parapen 0.1 Forischoline 0.05 Palm oil tocotrienol 0.5 B Water phase part Hyaluronic acid 0.2 Methylparaben 0.2 Purified water Balance C Fragrance Suitable amount ─────── ──────────────────────────── Total 100.0% The above formulations A and B were mixed and dissolved at 70 ° C. A was added and emulsified uniformly. Add C and cool,
An emulsion was prepared.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI A61K 45/08 A61K 45/08 A61P 17/00 A61P 17/00 43/00 107 43/00 107 (56) References 63-304978 (JP, A) JP-A-3-188024 (JP, A) International Publication 94/027583 (WO, A1) (58) Fields investigated (Int.Cl. ⁷ , DB name) A61K 35/78 A61K 7/00 A61K 31/355

Claims (3)

(57) [Claims] 1. An anti-aging agent comprising a root extract of Choleus forskolini and tocotrienol as active ingredients. 2. A bioaging agent containing forskolin and tocotrienol as active ingredients. 3. A skin composition containing the bioaging agent according to claim 1 or 2.