JP2953529B2 - New cosmetics - Google Patents

Description

The present invention relates to a novel bio-based cosmetic.

[Conventional technology]

With conventional cosmetics, it is difficult to remove dirt from the epidermis, it is difficult to remove bacteria while it is attached, excessive loss of necessary water and oil, low persistence in the epidermis, makeup collapse occurs immediately, repeatedly It is necessary to apply. Problems such as poor feeling of use such as stickiness and hot flash, discoloration and odor are mentioned. In addition, when it is desired to mix a functional component, an excessive amount of the functional component is mixed in terms of permeability and reach to skin cells.

Dirt on bare skin leads to a decrease in skin physiology and metabolism, hasten aging, irritates the skin, inflammation and bacterial infection,
It can cause allergies and can cause rough, moist, firm and dull skin. For this purpose, it removes bacteria and chemical substances attached to skin membranes, removes aged keratin, fat and proteinaceous dirt, replenishes lost water and nutrients, and adds color and fragrance to skin, nails and hair In addition, cosmetic materials such as proteins, polysaccharides and enzymes have been developed for the purpose of pursuing more beauty.

[Problems to be solved by the invention]

However, among these conventional cosmetics, those using antibodies in addition to collagen, hyaluronic acid, chondroitin sulfate, enzymes and the like are not known.

Antibodies are well known for their physiological functions, such as the ability to recognize and specifically bind antigens and the ability to activate the system that removes antigens, but no cosmetics that make use of their protein properties are found. . Further, the modified antibodies themselves or cosmetics containing them are not known.

[Means for solving the problem]

In general, the present invention relates to a cosmetic, which is characterized by containing an anti-keratin antibody and / or a modified antibody thereof.

The present inventors have solved the above problems with an antibody having specific properties and / or a dye, a fragrance, a humectant, a deodorant, a bactericide, a bacteriostat, an antioxidant, a surfactant, By forming a complex with a functional molecule such as an enzyme, the moisturizing property of the antibody itself is effectively given, and it is considered that the functional additive may be effectively transported and bound to the skin. .

As a result of earnest research based on the above ideas,
It has been found that the antibody binds to a target substance in the skin and exhibits moisturizing properties, and furthermore, that the antibody modified with the functional molecule binds to the epidermis and the target substance to exhibit a function, and the present invention completed.

Hereinafter, an anti-keratin antibody will be described more specifically as an example of the present invention.

The skin epidermis is only 0.2 mm thick and the outermost layer is composed of 4 to 5 layers. The outermost layers are the sebum and stratum corneum, preventing water from evaporating, preventing the invasion of various bacteria, preventing external stimuli, and covering dirt. Plays a role. This sebum film is formed by emulsification of moisture from sweat and oil of sebum on the epidermis, and is an ultrathin film of 0.5 μm, which has a function of protecting the stratum corneum and simultaneously holding dirt on the epidermis.
In addition, the stratum corneum is a hard protein called keratin, which is also present in hair, nails and the like. These keratins are high molecular weight proteins having a crosslinked structure and a molecular weight of 40,000 to 68,000, and are stable proteins that are insoluble in water. It is this sebum and stratum corneum that are greatly involved in the beauty of the skin.

The anti-keratin antibody described herein is an antibody obtained by using keratin of epidermal cells as an antigen and specifically binding to the antigen. Usually, if a rabbit, goat, sheep or other animal is hyperimmunized, a specific antibody against the antigen can be obtained from the serum. If the antigen is hyperimmunized to a chicken, a chicken egg antibody that shows specificity for the antigen can be obtained from eggs produced by the chicken.

Furthermore, a monoclonal antibody that specifically binds to the antigen, which is obtained by a fusion cell of an antibody-producing cell specific to the antigen and a myeloma cell, may be used.

Among them, a method of obtaining a specific antibody from a chicken egg using a chicken is desirable from an economical point of view and a sensory point of view. It is more practical to use specific antibodies obtained from chicken eggs than specific antibodies obtained from serum or cell culture, but this does not limit the present invention.

The functional compound used in the modified antibody in the present invention is a compound that maintains healthy physiological function of skin and maintains and promotes healthy beautiful skin. That is, skin components such as polysaccharides such as hyaluronic acid, collagen, and chondroitin sulfate, vitamins and amino acids that maintain and promote the normal production of these skin components are removed to prevent contamination of the skin surface from various bacteria. Antibacterial agents, bacteriostats, etc., and anti-inflammatory agents, hemostatic agents, anti-allergic agents, UV-blockers, melanin deposition inhibitors, etc. to prevent skin damage and hypersensitivity reactions due to external irritation There are various enzyme agents and the like, polysaccharides and the like for imparting moisturizing properties, pigments for coloring skin, and fluorescent pigments. As these functional compounds, those derived from nature and even foods are desirable from a sensory point of view.For example, phycobililine obtained from seaweed as a fluorescent dye is obtained from seaweed as a bacteriostat, a virus-static agent, and a humectant. Fucoidan.

Examples of the method for modifying an antibody in the present invention include covalent bonds and ionic bonds. Not only methods for modifying antibody proteins used in the field of conventional immunology are applicable, but also widely used in the field of biochemistry. Modification methods for proteins, sugars, nucleic acids, and the like can be used. The antibodies and / or modified antibodies themselves or substances containing them are applied to the skin, nails, hair and body hair in various forms.

For example, the anti-keratin antibody-containing egg yolk is mixed with a carrier generally used for the production of a composition applied directly to the skin and the like,
A composition to be applied to the skin or the like can be produced. Suitable such carriers include gels, liquid sprays,
Examples include creams, lotions, oils, powders, ointments, solid stick applicators, and the like.

For example, when an anti-keratin antibody-containing egg yolk is used as the antibody, the preparation usually contains about 1 to 25 parts by weight of egg yolk in a cosmetic carrier. The compounding amount of the antibody and / or the modified antibody in the present invention is preferably 0.0001 to 5 parts by weight.

When the amount is less than 0.0001 part by weight, the action of the antibody and / or the modified antibody in the cosmetic may not be sufficient, and even when the amount exceeds 5 parts by weight, an effect corresponding to the compounding agent may not be expected. However, the concentration range of the antibodies and / or modified antibodies in different compositions must be set for each formulation depending on the amount that produces the desired effect.

The novel cosmetics of the present invention can be applied to, for example, lotions, emulsions, creams, packs, bath preparations, facial cleansers, bath detergents, hair preparations, hair cleansers, and the like.
Skin cream, skin milk, cleansing cream, cleansing milk, cold cream, cream soap, makeup base, skin lotion, pack, calamine lotion, milky lotion, hand cream, essence powder, whitening powder, powder soap, solid soap, transparent Soap, lip balm, lipstick, nutritional essence, creamy foundation, face powder, powder eyeshadow, powder foundation, nail remover, hair tonic, hair liquid, hair cream, hair treatment, scalp treatment, shampoo, rinse, hair spray, sun oil , Sunscreen, shaving home, shaving cream, baby oil etc. That.

The cosmetics of the present invention may appropriately contain pigments, fragrances, preservatives, surfactants, pigments, antioxidants, and the like as long as the object of the present invention is achieved.

According to the cosmetic of the present invention, smoothness and the like are imparted to the skin by skin roughness improvement, keratin improvement, beautiful skin effect, etc., and to hair, excellent gloss is imparted, smooth feel, A finishing effect or the like can be provided.

〔Example〕

Next, the present invention will be described in more detail by way of examples. Note that the present invention is not limited to this. Hereinafter, in each example, parts and% are based on weight.

Example 1 Production of Egg Yolk Containing Anti-Keratin Antibody 1 ml of physiological saline solution containing 0.5 mg of keratin derived from human skin
1 ml of FCS (Freund's complete adjuvant) was mixed 1: 1 to form an emulsion. The resulting emulsion was injected into the pectoral muscle of chicken in 1 ml increments, and after the first immunization, 8 weeks later, the second immunization was carried out in the same manner as above, and after 1 month, eggs produced by the chicken were collected for 3 weeks. Eggs were obtained and egg yolk was obtained.

Example 2 Preparation of Antibody An additional 4.4% was added to a mixed solution obtained by mixing 500 ml of the above egg yolk and the same amount of PBS (phosphate buffered saline, pH 7.4).
Then, polyethylene glycol 6000 is added thereto, and the mixture is stirred at room temperature for 30 minutes, and centrifuged at 4 ° C., 6000 × g for 30 minutes to obtain a supernatant. The supernatant was passed through a phenyl sepharose (Pharmacia) column equilibrated with 1/10 PBS, the adsorbed fraction was eluted with distilled water, and the first peak was collected to obtain a purified egg yolk antibody (100 mg).

(1) A lotion was prepared from the following components.

Keratin antibody 0.1 part Ethanol 8.0 part 1,3-butylene glycol 2.0 part Methyl paraoxybenzoate 0.12 part Purified water residue (2) Prepared without blending keratin antibody with the lotion obtained in (1) (purified water replacement) Using the control lotion and the control lotion, a rough skin improvement effect test, a keratin improvement effect test, and a skin improvement effect test after tanning were performed on a human body panel using them. The results are shown in Tables 1 to 3.

(Surface Roughness Improvement Effect Test) The effect of continuous application for four weeks was examined on 20 middle-aged and elderly subjects having rough skin on the lower leg. Approximately 1 g of a sample was applied to the test site of the left lower leg of the subject once a day, and the skin condition before and after the start of the test was determined according to the following criteria. The lower right leg served as a control without the application of the sample.

* Evaluation criteria for skin dryness Evaluation points:-Normal ± Slightly dry, no desquamation + Dry, mild desquamation ++ Dry, moderate desquamation +++ Dried, desquamation Compare the test results before and after the test with the control site,
When skin dryness is improved by two stages (for example, + →-, +
(+ → ±) was effective, and the case where the level was improved by one step was regarded as slightly effective, and the results were shown by the number of subjects.

(Keratin improvement effect test) A mending tape was adhered to the test subject's skin before the start of the rough skin improvement effect test and after the test was finished, and when the mending tape was peeled off, the state of the keratinocytes attached to the tape was examined in detail by a scanning electron microscope. Examined. The anti-peeling properties of skin keratin were analyzed according to the following criteria, and the keratin-improving effect was determined.

* Evaluation criteria for keratin improvement effect (increase in exfoliation of keratin) Evaluation points: 1. No scale recognized 2. Small scale scattered 3. Small to medium scale remarkable 4. Large scale remarkable Evaluation was made after continuous application for 4 weeks The case where the difference between the evaluation of the test site and that of the control point was 2 or more was valid, the case of 1 point was slightly valid, and the case of 0 point was invalid. The judgment result is valid,
It is indicated by the number of subjects who became slightly effective.

(Skin improvement effect test after tanning) The above-mentioned Example 1- (1) was applied to the left and right skin of the forearm flexion side of 25 subjects irradiated with sunlight for 6 hours (3 hours a day for 2 days). The obtained lotion and a control lotion containing no antibody were applied twice a day to each of the left and right sides for 10 days, and the effect of improving skin roughness was examined.

On the tenth day, the keratin-improving effect was evaluated on the 10th day according to the same criterion.

Example 3 Preparation of Fluorescent Modified Antibody Phycobiliprotein was bound to the yolk antibody prepared in Example 2 by the SPDP method. Specifically, 1 μmol of phycobiliprotein was dissolved in 2 ml of 0.1 M sodium phosphate buffer (pH 7.5) containing 0.1 mM NaCl, and 20 mM N-succinimidyl 3- (2-pyridyldithio) propionate (SPD
P) 0.1 ml of an ethanol solution was added, and the mixture was subjected to gel filtration with Sephadex G-25 (Pharmacia) left at room temperature for 30 minutes.
SPDP-modified phycopyrin protein was obtained. Next, 1 μmol of antibody
Was subjected to SPDP in the same manner as described above, and dithiothreitol was added thereto to a final concentration of 50 mM. The mixture was allowed to stand at room temperature for 30 minutes, and subjected to gel filtration with Sephadex G-25 to obtain an SH-conjugated antibody. This is mixed with the SPDP-modified phycobiliprotein and mixed at room temperature.
The mixture was reacted overnight and subjected to gel filtration using Ultrogel AcA22 (manufactured by LKB), and the first peak was collected as a fluorescent dye-modified antibody.

(Color coating effect test) The fluorescent labeled antibody obtained in Example 3 was
It was mixed with talc so as to contain 1% as phycobiliprotein. A 0.1 g sample containing a fluorescently labeled antibody equivalent to 1 mg of phycobiliprotein was applied to the left arm of 10 panelists. On the other hand, as a control, a talc preparation containing the same amount of only phycobiliprotein not bound with an antibody was applied to the right arm of the same panelist in the same amount. Twelve hours later, an excitation wavelength of 545 nm was applied to the left and right arms, and the intensity of the fluorescence was visually determined. As a result, in all ten persons, the left arm to which the phycobiliprotein bound to the antibody was applied emitted stronger fluorescence.

This result indicates that the color application effect is increased and the effect is persistent by binding the fluorescent substance to the anti-keratin antibody and applying it to the skin.

Example 4 Preparation of polysaccharide-modified antibody Fucoidan was bound to an anti-keratin antibody by a periodate oxidation method.

10 mg of fucoidan was dissolved in 1 ml of distilled water, 0.2 ml of 0.1 M sodium periodate was added, reacted at room temperature for 20 minutes, and dialyzed against 1 mM sodium acetate buffer (pH 4) overnight. to this
Add 0.02 ml of 0.2 M sodium carbonate buffer (pH 9.5) to pH 9
At the same time as 9.59.5, 10 mg of antibody was added and reacted at room temperature for 2 hours. Sodium borohydride (4mg / ml)
0.1 ml was added thereto, and the mixture was reacted at 4 ° C. for 2 hours. The resulting mixture was subjected to gel filtration with Ultrogel AcA22, and the first peak was collected as a polysaccharide-modified antibody.

(1) An emulsion was prepared from the following components.

Liquid paraffin 23.0 parts Isopropyl myristate 5.0 parts Vaseline 6.0 parts Beeswax 5.0 parts Stearic acid 2.0 parts Behenyl alcohol 1.0 parts Polysorbetane monostearate 3.0 parts Polyoxyethylene (20) sorbitan monostearate 3.0 parts 1,3-butylene glycol 3.0 parts Paraben 0.3 Part fucoidan-modified keratin antibody 0.1 part flavor appropriate amount purified water residue Emulsion containing 1% fucoidan in place of the emulsion obtained in (1) and fucoidan-modified keratin antibody, and purified water instead of fucoidan-modified keratin antibody as a control emulsion A skin effect test was conducted in comparison with the compounded product.

(Beautiful skin effect test) Female subjects complaining of rough skin, fine wrinkles, dry skin, etc. (35-
55 years old) Apply the sample to 20 people twice a day, morning and evening for 3 consecutive months.
Immediately thereafter, the following items were evaluated.

* Evaluation items Skin wettability: Number of people who answered that the skin was moist Sticky: Number of people who answered that the sample felt sticky Skin smoothness: Number of people who answered that the skin became smooth Elasticity: Skin tension Number of people who answered that irritation occurred Irritation student: Number of people who felt irritation at the time of application Irritation: Number of people who answered that the skin was rash during use period The results are shown in Table 4 below.

Example 5 Cream Liquid paraffin 23.0 parts Isopropyl myristate 5.0 parts Vaseline 6.0 parts Beeswax 5.0 parts Stearic acid 2.0 parts Behenyl alcohol 1.0 parts Sorbitan monostearate 3.0 parts Polyoxyethylene (20) sorbitan monostearate 3.0 parts 1,3-butylene glycol 3.0 parts Paraben 0.3 parts Anti-keratin 0.5 parts Purified water Residue Example 6 pack Polyvinyl alcohol 15.0 parts (Saponification degree 88% polymerization degree 2000) Polyethylene glycol 3.0 parts Propylene glycol 7.0 parts Ethanol 10.0 parts Methyl paraben 0.1 parts Fucoidan-modified anti-keratin antibody 1.0 Part Perfume Appropriate amount Purified water Residue Example 7 Hair tonic Propylene glycol 3.0 parts Pantothenol ethyl ether 1.0 parts Diphenhydramine hydrochloride 0.1 parts Salicylic acid 0.1 parts 1-menthol 0.1 parts Fucoidan-modified anti-keratin antibody 0 .5 parts Ethanol 40.0 parts Perfume Appropriate amount Purified water residue Example 8 Lip Treatment Candela wax 9.0 parts Solid paraffin 8.0 parts Beeswax 5.0 parts Carnauba wax 5.0 parts Lanolin 11.0 parts Castor oil Residue isopropyl myristate 10.0 parts Perfume Appropriate amount Mucopolysaccharide-modified keratin antibody 0.1 part Example 9 Hair rinse Stearyl trimethylammonium chloride 4.5 parts Stearic acid 0.5 parts Ethanol 1.0 parts Monostearic acid 1.0 parts Polyoxyethylene oleyl ether 3.5 parts 1,3-butylene glycol 3.0 parts Fragrance 0.1 part Anti-keratin antibody 0.2 parts Purified water Residual Example 10 Bath agent [Effects of the Invention] According to the present invention, there is provided a novel cosmetic that effectively transports a functional substance to the skin, has excellent functional persistence, and is excellent in mass productivity and safety.

──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Ikuyuki Kato 3-4-1, Seta, Otsu City, Shiga Prefecture Inside Takara Shuzo Co., Ltd. (56) References JP-A-1-313439 (JP, A JP-A-2-121908 (JP, A) JP-A-2-121909 (JP, A) JP-A-2-121910 (JP, A) JP-A-60-8217 (JP, A) US Pat. , A) British Patent No. 1585661 (GB, B) British Patent No. 1247886 (GB, B) WO 89/11866 (WO, A1) Encyclopedia of Biotechnology (1986, published by CMC Co., Ltd.) 1058-1060 Iwanami Biology Encyclopedia (1969, published by Iwanami Shoten Co., Ltd.), 289-290, Cell, Vol 30, 361-372 Nisshinkai; 95-11 (1985), 1153-1157 (58) Fields investigated (Int. Cl. ⁶ , DB name) A61K 7/00 A61K 39/395

Claims (4)

(57) [Claims] 1. A cosmetic comprising an anti-keratin antibody and / or a modified antibody thereof. 2. The cosmetic according to claim 1, wherein the antibody is a chicken egg antibody. 3. The cosmetic according to claim 1, wherein the keratin is keratin of epidermal cells. 4. The cosmetic according to claim 1, which is a lotion, an emulsion, a cream, a pack, a bath agent, a face wash, a bath detergent, a hair agent or a hair wash.