JP2006316050A - Anti-wrinkle agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new anti-wrinkle agent, a stabilizer, etc., for basement membranes. <P>SOLUTION: The new anti-wrinkle agent comprising any one or more of extracts from Nasturtium officinalis R. Brown, Juniperus oxycedrus Linne, Atractylodes macrocephale and Tussilago farfara Linne as an active ingredient is provided. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a novel anti-wrinkle agent. More specifically, the present invention relates to an anti-wrinkle agent comprising, as an active ingredient, an extract derived from Dutch pepper, pepper, peanut and / or dandelion. In addition, the present invention provides a basement membrane stabilizer, an extracellular matrix production promoter and skin sagging, a firmness improving agent, a method for producing them, and a production method for artificial skin using the extract. The present invention relates to a method, a method for reducing wrinkles, sagging skin, and a method for improving elasticity.

  Skin covering the entire body of various animals including human beings is exposed to external factors such as sunlight, dryness, oxidation, environmental stress, mental stress, and wrinkle formation due to aging, hardening, spots, dullness, elasticity It is exposed to changes such as decline. Here, the skin is roughly divided into two layers, the epidermis and the dermis. Between the epidermis and the dermis there is a thin and delicate membrane called the basement membrane. Epidermal metabolism depends on factors and blood supply produced by dermal cells through this basement membrane. Epidermal growth and differentiation in the skin is regulated by the basement membrane and dermis. Therefore, the communication between the epidermis and dermis via the basement membrane plays an important role in regulating the function of the skin epidermis.

  The skin basement membrane has a special structure called an anchoring complex, which plays a role in stabilizing the adhesion and communication between the two tissues, the epidermis and dermis. The anchoring complex proteins are linked to both keratin cytoskeleton of keratinocytes and connective tissue proteins of the dermal papillary layer. One important component of the anchoring complex is laminin 5 (Rousselle et al., J Cell Biol. 1991 Aug; 114 (3): 567-76). Previous studies have revealed that genetic variation in the gene encoding the three strands of laminin 5 is responsible for the congenital genetic disease that forms severe blistering, Herritz junctional epidermolysis bullosa. (Aberdam et al., Nat Genet. 1994 Mar; 6 (3): 299-304). This indicates that laminin 5 is an essential substance for adhesion of the epidermis. In addition, laminin 5 forms an anchoring filament and has been elucidated to bind to integrin α6β4 (a transmembrane integrin present in hemidesmosome) known as a receptor for laminin 5 (Niessen). et al., Exp Cell Res. 1994 Apr; 211 (2): 360-7).

  Laminin 5 not only binds directly to type VII collagen, which forms anchoring fibers connected to the dermal papillary layer, but also forms a complex with other laminin 6 and 7, which is responsible for naidogen. It is reported that it binds to type IV collagen, which is the skeleton of the basement membrane.

  The expression level of type IV collagen constituting the complex has been observed to decrease with aging (Vazquez F et al., Maturitas 1996, 25: 209-215), and type VII collagen to which laminin 5 binds. As for dermal fibroblasts derived from elderly people, the production ability is reduced at the protein level and mRNA level compared to dermal fibroblasts derived from young people (Chen et al., J. Invest. Dermatol., 102: 205-209, 1994). In addition, anchoring fibers composed of type VII collagen are reported to decrease in normal skin with physiological aging and photoaging (Takuo Tsuji, Nisshinkai 105: 963-975, 1995, Tidman et al., J Invest Dermatol., 83: 448-453, 1984). In addition to the characteristics of these individual components, the basement membrane itself is known to exhibit more than a structure such as multiplexing and rupture with skin aging (Lavker et al., J. Invest. Dermatol. 1979 73: 59-66), as a result of this structural change, it seems that the appearance of senile features such as wrinkles and sagging, and a decrease in skin function associated with aging. Therefore, promoting the production of laminin 5, type IV and type VII collagen constituting the basement membrane skeleton is considered to be extremely important in maintaining the basement membrane structure in a good state and preventing skin aging. In fact, when the skin basement membrane structure is observed in detail, basement membrane damage is frequently observed in the late 20s and early 30s, indicating that the structural change of the basement membrane plays an important role in the induction of skin aging. (IFSCC Magazine, 2000 4 (4): 15-23).

  Anti-wrinkle is one of the major problems in preventing skin aging. Here, wrinkles are roughly classified into shallow epidermal wrinkles (small wrinkles) and deep dermal wrinkles (large wrinkles). In particular, dermal wrinkles are caused by changes in the basement membrane and the dermal layer. It is thought that the maintenance of the basement membrane is important for this reason.

  Furthermore, in addition to the above-mentioned basement membrane components, type I and type III collagen (about 80% and about 15%, respectively), which are present in large amounts in the dermis, decrease in absolute amount in the dermis with aging, It is believed that these changes are likely related to reduced dermal elasticity and firmness, wrinkles, or dermal atrophy. Focusing on type III collagen, the ratio of type III collagen is higher in fetal to infant tissues than in adult tissues, and in Erasdanros syndrome type IV, where wrinkles are conspicuous, there is no type III collagen in the tissue. It is considered that it is likely to be an important ingredient in building soft skin.

  Here, type I and type III collagen in the dermis is synthesized by fibroblasts. When this fibroblast is cultured in a collagen gel, the collagen gel contracts. The fiber density of the contracted collagen gel is close to the fiber density in the dermis. This is a phenomenon that occurs because fibroblasts attract collagen fibers. Even in actual skin, it is considered that the elasticity of the skin is maintained by fibroblasts sufficiently attracting collagen fibers. Thus, in addition to the basement membrane, maintaining the dermis is also important for maintaining skin homeostasis, such as anti-wrinkle, skin sagging, and improvement of skin.

  So far, preparations derived from soybeans, lysophosphatidylcholine, lysophosphatidic acid for the purpose of enhancing the ability to produce laminin 5, the main component of the basement membrane, from the viewpoint of maintaining skin homeostasis Furthermore, extracts of plants belonging to the genera Bacconia, Psophocarpus, Cassia, and Erythraea have been proposed (Japanese Patent Laid-Open Nos. 11-343226 and 2000-226308). And JP-A-2003-313135). Similarly, with regard to type III, type IV, and type VII collagen production promoters, various plant-derived extracts such as soybeans, beech buds, cuckoos, western ivy, and beech family beech extracts are known. (JP 2001-39849, JP 2004-18471, JP 2004-75661, etc.). However, no other substance is known to promote the production of laminin 5, type IV, type VII collagen, which is an important component of the basement membrane, and type III collagen, an important component of the dermis. (JP 2001-39849, JP 2003-137767, and JP 2004-18471).

JP 2001-181120 A JP 2003-313113 A JP 2001-288066 A JP 2003-194809 A JP 2004-161639 A JP 2002-12548 A

  The present invention relates to the provision of a novel anti-wrinkle agent.

  As a result of intensive studies on various plant extract materials with the aim of obtaining an anti-wrinkle action especially against dermal wrinkles, the present inventor is a key component of the basement membrane. It significantly promotes the production of laminin 5, type IV collagen, type VII collagen, and type III collagen, an important component of the dermis, and when applied to the skin, it also has an anti-aging effect on skin, particularly an anti-wrinkle effect and skin It became clear that sagging and firmness were improved. At the same time, it was also found out that, among the above extracts, pepper, peanut and dandelion have an anti-inflammatory action and a wound healing action. So far, no anti-wrinkle action has been reported in skin aging, including the promotion of production of extracellular matrix including laminin 5, with respect to the plant extract.

  Dutch Rape (scientific name: Nasturtium officinalis R. Brown): An extract of Dutch Rape, a kind of Brassicaceae, has a blood circulation promoting action (Japanese Patent Laid-Open No. 2001-181120) and is used as a hair growth agent by promoting the production of vascular endothelial growth factor (specialized) Open 2003-313113) is known. However, the effect on skin aging is not known.

  Tomato (Juniperus oxycedrus Linne): An extract of tomato which is a kind of cypress family is known to have an effect of improving skin barrier function such as rough skin (Japanese Patent Laid-Open No. 2001-288066), but skin aging, anti-wrinkle, skin There is no known effect on sagging or elasticity. The skin barrier function improving effect referred to here is for the stratum corneum and does not mention any effect on the dermis below the stratum corneum.

  Sandalwood (scientific name: Atractylodes macrocephale): An extract of sandalwood, which is a member of the Asteraceae family, is known to have anti-skin roughening action (JP 2003-194809) and anti-white hair action (JP 2004-161639), There are no known anti-wrinkle, skin sagging or elasticity improvements.

  Fusdanpopo (scientific name Tussilago farfara Linne): An extract of Fusdanpopo, a member of the family Asteraceae, has a phototoxicity-inhibiting effect, for example, an action that prevents aging due to products generated by UVA irradiation of air pollutants (Japanese Patent Laid-Open No. 2002-12548) However, the prevention / amelioration action against wrinkles mainly caused by UVB irradiation and the skin sagging and firmness improvement actions are not known.

  Accordingly, the present invention provides an anti-wrinkle agent, a basement membrane stabilizer, an extracellular matrix production promoter, and a skin, which comprise any one or more of the extracts of Dutch pepper, pepper, peanut and dandelion as an active ingredient. The present invention provides a sagging agent, a firmness improving agent, and a production method thereof, a method for producing an artificial skin using the above extract, a wrinkle reduction and skin sagging, and a cosmetic method for improving the firmness.

  The anti-wrinkle agent according to the present invention contains extracts of Dutch pepper, pepper, sand jelly, and dandelion, so that laminin 5, type IV collagen, type VII collagen, and important dermis are important components of the basement membrane. Promotes the production of extracellular matrix such as type III collagen as a constituent. In addition, by promoting the production of extracellular matrix, the basement membrane and dermis are repaired, wound healing is achieved, the skin condition is normalized, such as reduced transdermal water transpiration, increased skin viscoelasticity, or Anti-wrinkle action is achieved, such as a reduction in the number and length of wrinkles. Although the exact mechanism is not clear, all of these actions are thought to be due to the restoration and stabilization of the basement membrane and dermis as a result of the accelerated production of laminin and the like. Therefore, the above extract also contributes to the improvement of symptom of epidermolysis bubbling caused by instability of basement membrane such as thawing of basement membrane cells, detachment of basement membrane from epidermis, type VII collagen abnormality, etc. Yes. In addition, it should be particularly noted regarding the effect of the present invention that all of the above-mentioned extracts promote the production of all of laminin 5, type III, type IV collagen and type VII collagen, not alone. .

Anti-wrinkle agent The anti-wrinkle agent of the present invention comprises an extract of Dutch pepper, pepper, sand bud and / or dandelion as an active ingredient. The term “anti-wrinkle” used in the present specification means an action to prevent and / or ameliorate skin wrinkles among the “skin aging preventive actions” described below, particularly to reduce the number and length of skin wrinkles. Is sometimes used synonymously with "skin aging prevention action". In addition, “wrinkles” that are improved / prevented by the present invention particularly mean deep dermal wrinkles caused by the depression of the dermis layer due to changes in the basement membrane. These effects are determined in consideration of factors such as improvement of keratin by visual sense determination, increase in skin moisture, and increase in skin elasticity. The above-mentioned effect achieved in the present invention is achieved by increasing the production amount of extracellular matrix such as laminin 5, type III, type IV or type VII collagen from the extract of Dutch pepper, pepper, sand squirrel and squirrel poppo. It is thought that this is due to stabilization of the dermis and improvement of skin homeostasis, but the exact mechanism is not clear. Therefore, it is clear that the anti-wrinkle action obtained by the anti-wrinkle agent of the present invention is not limited to that caused by an increase in the amount of extracellular matrix produced. In addition, although the prevention / improving action of skin aging or wrinkle production caused by UVA irradiation is known for the dandelion extract (Japanese Patent Application Laid-Open No. 2002-12548), the prevention / improving action against wrinkles mainly caused by UVB is unknown.

  As used herein, “skin aging prevention” refers to prevention of skin function deterioration due to the accumulation of structural changes in the basement membrane due to aging and photoaging, specifically wrinkles, sagging and hardening of the skin. Means to improve, maintain a youthful and healthy skin condition with elasticity.

Basement Membrane Stabilizer The present invention provides a basement membrane stabilizer comprising as an active ingredient any one or more of extracts derived from Dutch pepper, pepper, peanut and dandelion. As used herein, “basement membrane stabilization” refers to the basement membrane that exists between the epidermis and the dermis and is deeply involved in maintaining the homeostasis of the skin, and its skeleton is stabilized and further repaired. Is meant to be done. More specifically, it is intended to mean an overall effect on the basement membrane caused by the promotion of production of important components constituting the basement membrane, particularly the extracellular matrix.

Extracellular matrix production promoter The present invention further provides an extracellular matrix production promoter comprising any one or more of the above extracts as an active ingredient. As used herein, “extracellular matrix” refers to those generally recognized in the art, particularly in basement membranes including laminin 5, type IV or type VII collagen. It means important constituents and important constituents of the dermis such as type III collagen.

Skin sagging and firmness improving agent The present invention further provides an agent for improving skin sagging and firmness comprising any one or more of the above extracts as an active ingredient. As used herein, the term “skin sagging and firmness improvement” means that skin irritation and aging are improved by improving the elasticity of the skin and tightening. For example, it can be evaluated by the collagen gel contraction promoting action in vitro, and can be evaluated by skin viscoelasticity in vivo.

  In addition to anti-wrinkle action, basement membrane stabilization action, extracellular matrix production promotion action, skin sagging, firmness improvement action and collagen gel contraction promotion action, the above extract also has anti-inflammatory action and wound healing promotion action . However, as described above, the exact mechanism of these actions is not clear. As used herein, “anti-inflammatory effect” and “wound healing promoting effect” are relatively determined when the plant extract of the present invention is applied to an inflamed site and a wound site, respectively, compared to a non-applied site. It is what is done. The wound healing promoting effect obtained by the present invention is considered to be due to the stabilization and repair of the basement membrane as a result of the promotion of extracellular matrix production. However, it is clear that this action is not necessarily limited to that resulting from increased extracellular matrix production. Among the extracts used in the present invention, wound healing effects have already been reported for Dutch pepper extract (Japanese Patent Laid-Open No. 2003-313134).

  In addition to the above-mentioned anti-inflammatory action and wound healing promotion action, the plant extract used in the present invention also has an action to improve epidermolysis bullosa. As used herein, the “amelioration of epidermolysis bullosa” refers to alleviation of general epidermolysis bullosa classified into simple epidermolysis bullosa, junctional epidermolysis bullosa and dystrophic epidermolysis bullosa Intended not only to cure, but also to prevent the symptoms. This effect is brought about by stabilizing the structure of the basement membrane by promoting the production of collagen, which is an important component of the basement membrane.

  The plant extracts used in the present invention are derived from Dutch peppers, peppers, peanuts and dandelions. Extracted from the roots, leaves, stems, flowers, etc. of herbaceous Dutch peppers, peppers, peanuts and dandelions, and extracted from the roots, buds, bark, fruits, leaves, flowers, etc. Is done.

  The extraction method may be performed by solvent extraction. In the case of solvent extraction, the plant material is dried as necessary, and further shredded or pulverized as necessary, and then an aqueous extractant such as cold water, hot water, or It is extracted by using hot water having a boiling point or lower temperature, or an organic solvent such as methanol, ethanol, 1,3-butanediol, ether or the like at normal temperature or by heating. However, the extraction method is not limited to solvent extraction, and may be a conventional method known in the art. The form of the extract may include not only the extract itself, but also those that are appropriately diluted or concentrated or dried by conventional techniques.

  The anti-wrinkle agent, basement membrane stabilizer, extracellular matrix production promoter, skin sagging, and firmness improving agent of the present invention may contain each extract alone or in combination with a plurality of extracts. In yet another embodiment, the anti-wrinkle agent, basement membrane stabilizer, extracellular matrix production promoter or skin sagging / harness improving agent of the present invention may further comprise a serine protease inhibitor (serine protease inhibitor). Are you sure you want to completely delete the mode of adding the inhibitor (you can leave this sentence alone, aside from the description below)? As described in the examples below, the anti-wrinkle agent, basement membrane stabilizer, extracellular matrix production promoter or skin sagging and firmness improving agent of the present invention contain a serine protease inhibitor, thereby degrading collagen. Inhibits to keep the basement membrane and dermis in better condition. As a result, the anti-wrinkle action, skin sagging, firmness improving action, wound healing promoting action, epidermolysis bullosa improving action and the like obtained by the present invention are expected to be further improved. Examples of the serine protease inhibitor include, but are not limited to, extracts of mint mint, yew, hypericum, jujube, arnica, karin, hawthorn, sunflower, rose rose, wild rose, grape, button, hop, raspberry, roman chamomile These can be mentioned, and these can be extracted by solvent extraction or the like in the same manner as the above-mentioned extraction method such as Dutch pepper. Further, the serine protease inhibitors include aprotinin, tranexamic acid, ε-aminocaproic acid and the like.

  The above extract used in the present invention is an amount necessary for achieving an anti-skin aging effect, particularly an anti-wrinkle effect and skin sagging, and an effect of improving elasticity, or stabilization of the basement membrane and dermis or laminin 5, type III The amount necessary to achieve an increase in the production amount of extracellular matrix such as type IV or type VII collagen is added. Furthermore, the necessary amount is blended according to the purpose such as increasing the amount of water in the stratum corneum or transdermal (improving rough skin), increasing skin elasticity, improving skin texture, etc. It varies depending on various factors. Although not limited, the extract is a concentration of 0.0001 to 10% by mass with respect to the total mass of the anti-wrinkle agent, basement membrane stabilization, extracellular matrix production promoter or skin sagging, and elasticity improving agent, Preferably, it may be blended at a concentration of 0.001 to 1% by mass.

  The anti-wrinkle agent, basement membrane stabilizer, extracellular matrix production promoter or skin sagging and firmness-improving agent of the present invention are not limited to the above-mentioned extract, and are generally used in cosmetics and the like within the range not impairing the anti-wrinkle action. Ingredients that can be incorporated into pharmaceuticals, such as liquid fats, solid fats, waxes, hydrocarbons, higher fatty acids, higher alcohols, esters, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic Surfactant, moisturizer, water-soluble polymer compound, thickener, coating agent, UV absorber, sequestering agent, lower alcohol, polyhydric alcohol, saccharides, amino acids, organic amines, pH adjuster, skin Nutrients, vitamins, antioxidants, fragrances, powders, coloring materials, water, and the like can be appropriately blended as necessary. Further, L-ascorbic acid and its salts, L-ascorbic acid phosphate ester, ester derivatives of L-ascorbic acid such as L-ascorbic acid sulfate, and salts thereof, and L-ascorbic acid such as L-ascorbic acid glucoside Glycosides and salts thereof, alkoxysalicylic acids such as 4-methoxysalicylic acid and salts thereof, hydroquinone glycosides such as hydroquinone β-D-glucose and hydroquinone α-D-glucose and salts thereof, tranexamic acid and tranexam Tranexamic acid derivatives such as acid methylamide hydrochloride, resorcin derivatives such as 4-n-butylresorcin, kojic acid, ellagic acid, linoleic acid, chamomile extract, retinoic acid, retinol, retinol acetic acid, retinol palmitic acid, glycyrrhizic acid and derivatives thereof Etc., Rhizophosphatiji Lucolin, lysophosphatidic acid, laminin 5 production promoters such as soybean preparations, sugars such as glucose, fructose, mannose, sucrose, trehalose, nicotinic acid benzyl ester, nicotinic acid β-butoxyethyl ester, capsaicin, gingerone, Cantharis tincture, ictamol, caffeine, tannic acid, α-borneol, tocopherol nicotinate, inositol hexanicotinate, cyclandrate, cinnarizine, trazoline, acetylcholine, verapamil, cephalanthin, γ-oryzanol and other blood circulation promoters, sulfur, Antiseborrheic agents such as thianthol, for various purposes, hinokitiol, iris extract, asenyaku extract, allantoin, aloe extract, yakiyaku extract, ibujakaku extract, turmeric extract, o Extract, Auren extract, Hypericum extract, Ononis extract, Hydrolyzed casein, Hydrolyzed yeast extract, Cuckoo extract, Xylitol, Clara extract, Rice extract hydrolyzed solution, Psycho extract, Saffron extract , Zinc oxide, winged bean extract, lion extract, peony extract, ginger extract, silica-coated zinc oxide, sage extract, mallow extract, sensu extract, assembly extract, chimpi extract, chili extract, Toki extract, Tonin extract, thiotaurine, carrot extract, garlic extract, Hiragi extract, Hipotaurine, birch extract, loquat extract, grape extract, beech bud extract, loofah extract, marjoram extract, Lily extract, Yokuinin extract, Rosemary extract, Arginine and its hydrochloride, Amino acids and derivatives thereof, such as calcium and its hydrochloride, pantothenic acids such as calcium pantothenate, D-pantothenyl alcohol, pantothenyl ethyl ether, acetyl pantothenyl ethyl ether, vitamin Ds such as ergocalciferol and cholecalciferol, nicotine Nicotinic acids such as acid, nicotinic acid amide and benzyl nicotinate, vitamin E such as α-tocopherol, tocopherol acetate, DL-α-tocopherol nicotinate, DL-α-tocopherol succinate, vitamin K, vitamin P, biotin, etc. Vitamins, coenzymes such as ubiquinone, and the like can be appropriately blended.

  The dosage form of the anti-wrinkle agent, basement membrane stabilizer, extracellular matrix production promoter or skin sagging agent, and firmness improving agent of the present invention is not particularly limited, and examples thereof include solution systems, solubilization systems, and emulsion systems. , Powder dispersions, water-oil two-layer systems, water-oil-powder three-layer systems, ointments, gels, aerosols and the like can be used. Further, the usage form is not particularly limited, and can take any form such as lotion, emulsion, cream, essence, jelly, gel, ointment, pack, foundation, and the like.

Skin external preparation Furthermore, this invention provides the skin external preparation containing the said extract. Here, the external preparation for skin of the present invention is used for the purpose of anti-wrinkle action and skin sagging, anti-skin aging action excluding firmness improvement action, anti-inflammatory action, wound healing promotion action, epidermolysis bullosa amelioration action, etc. . However, the external preparation for skin of the present invention is not limited to these purposes, and use intended to improve various symptoms of the skin caused by the action of promoting the production of extracellular matrix by the above extract can be considered.

  Substances to be blended in the external preparation for skin of the present invention are the same as those described above for the anti-wrinkle agent and the like, but the type, amount, concentration, etc. are appropriately determined by those skilled in the art according to the desired purpose. sell. For example, when aiming at an anti-inflammatory action and a wound healing promotion action, the extract is at a concentration of 0.0001 to 10% by mass, preferably 0.001 to 1% by mass, with respect to the mass of the whole external preparation for skin. May be blended at a concentration of

In another embodiment of the method for producing an anti-wrinkle agent, a basement membrane stabilizer, an extracellular matrix production promoter or skin sagging, a firmness-improving agent , the present invention relates to an extract derived from Dutch pepper, pepper, peanut and dandelion in an aqueous phase or Provided is a method for producing an anti-wrinkle agent, a basement membrane stabilizer, an extracellular matrix production promoter or skin sagging, and a firmness improving agent, comprising a step of adding to an oil phase. This method can also be applied to the above-mentioned external preparation for skin.

Method for producing artificial skin Further, the present invention provides a method for producing artificial skin, characterized in that any one or more of the extracts of Dutch pepper, pepper, peanut and dandelion are added to the artificial skin formation medium. ,I will provide a.

  As the basal medium used for the production of artificial skin in the present invention, any medium conventionally used for the production of artificial skin can be used, and these media include Dulbecco's modified eagle containing 10% fetal calf serum. Medium (DMEM): DMEM-Ham'sF12 (3: containing 10% fetal bovine serum, transferrin 5 μg / ml, insulin 5 μg / ml, tri-iodothyronine 2 nM, cholera toxin 0.1 nM, hydrocortisone 0.4 μg / ml 1); a medium in which keratinocyte growth medium (KGM) and DMEM containing 10% fetal bovine serum are mixed at 1: 1, and the like. Although the said extract added to these basal media changes with the kinds, it is about 0.001 mass%-about 1 mass%. In addition to the extract, various substances used in the embodiment of the anti-wrinkle agent may be added together. In particular, the above-mentioned serine protease inhibitor inhibits collagen degradation and stabilizes the basement membrane. It is considered preferable.

In the production of the artificial skin of the present invention, first, a contracted type I collagen gel containing human fibroblasts is placed on a wire mesh. A contracted type I collagen gel containing human fibroblasts can be prepared, for example, as follows. After preparing a fibroblast-suspended collagen solution on ice, collagen is gelled in a Petri dish. Thereafter, the gel is peeled from the wall surface of the Petri dish, and the collagen gel is contracted in a CO ₂ incubator.

Next, epidermal cells such as human normal epidermal keratinocytes are cultured on the collagen gel to form the epidermis. Formation of the epidermal layer by culturing skin cells can be performed as follows. A contracted collagen gel is placed on a wire mesh, and a glass ring is placed on the gel. An epidermis keratinocyte suspension derived from human foreskin is placed in the glass ring so as not to leak. Adhere keratinocytes in a CO ₂ incubator and remove the ring. The medium is filled up to the boundary of the epidermal layer, and the culture is continued while the epidermal layer is exposed to air to form a stratum corneum.

  According to this method, artificial skin close to natural skin in which a basement membrane component is sufficiently deposited between the dermis layer composed of contracted type I collagen gel containing fibroblasts and the epidermis layer can be obtained. The extract used in the present invention is not only contained in a new medium at the time of medium change on the first few days after seeding of epidermal cells, but is not limited to this stage, and may be added as needed. Can do.

In another aspect of the method for reducing wrinkles and sagging skin , the present invention provides a method for reducing wrinkles by applying to the skin any one or more of the extract of Dutch pepper, pepper, sand bud and dandelion. And a cosmetic method characterized by improving skin sagging and firmness. The method not only reduces wrinkles, sagging skin, and improves skin tension, but also promotes the production of extracellular matrix as described above, and various skin symptoms that require stabilization of the basement membrane and dermis, For example, it is intended for use to improve epidermolysis bullosa. In this method, the subject to which the above extract is applied is generally a mammal, and can be applied to humans in particular.

  Next, the present invention will be described more specifically with reference to the following examples.

(Experiment Method-1)
Test method for laminin 5 production promoting effect (1) Culture of epidermal keratinocytes Epidermal keratinocytes were isolated from human foreskin and cultured in epidermal cell growth medium (KGM) with low calcium concentration. The bovine pituitary extract and EGF were added to this medium. The cells were cultured in KGM until the fourth generation, then the adherent cells were suspended by trypsin-EDTA treatment, the cell aggregates were removed by filtration, and a uniform cell suspension was obtained. Cells were collected by centrifugation and resuspended in DMEM-F12 (2: 1) -0.1% BSA to 8 × 10 ⁴ / ml. This cell suspension was added to 0.5 ml of 0.5 ml of the same medium containing a double concentration of the drug. The culture was performed at 37 ° C. for 24 hours using a 24-well plate. At the end of the culture, the culture supernatant was transferred to an Eppendorf tube, centrifuged at 10,000 rpm for 5 minutes, the supernatant transferred to a new tube, and stored at −20 ° C. until the day of laminin 5 measurement. In addition, in order to solubilize laminin 5 bound to the inside of the cell and the culture plastic, Tris-HCl buffer (pH 7.4) containing various surfactants was added to each well and stored at −20 ° C. overnight. The next day, it was sonicated and frozen again. The next day, after dissolving again, the mixture was centrifuged at 10,000 rpm for 5 minutes, and the supernatant was transferred to a tube and stored at −20 ° C. until the day of measurement of laminin 5.

(2) Measurement of laminin 5 by sandwich ELISA method Laminin 5 present in the culture supernatant and cell layer was measured by sandwich ELISA method. A BM165 monoclonal antibody against laminin α3 chain of laminin 5 was bound to a solid layer of a 96-well ELISA plate. In order to sandwich and measure laminin 5, 6F12, which is a monoclonal antibody against laminin β3 chain, was biotinylated (b-6F12) in advance as another type of antibody. In this method, only the heterotrimeric body (α3β3γ2) capable of exhibiting the function is measured, and the heterodimer (β3γ2) is not detected. Samples are added to each well previously filled with 3% gelatin / phosphate buffer solution containing b-6F12. The final dilution ratio in the sample hole was 1/4 for the culture solution and 1/10 for the cell layer. The antigen-antibody reaction was carried out at 37 ° C. for 2 hours. After washing the plate, an avidin HRP (horseradish peroxidase) solution was added, and the reaction was further continued for 30 minutes to 1 hour. After washing, an ABTS solution which is a substrate for HRP was added, and the absorbance at 405 nm was measured with an ELISA plate reader. A calibration curve was prepared in the range of 0 to 40 ng / ml.

  The amount of laminin 5 produced was calculated as the sum of the amount released in the medium and the amount remaining in the cell layer, and indicated as a relative value to the sample (control) to which no plant extract was added. Table 1 below summarizes the rate of increase in laminin 5 production for each extract.

(Experiment Method-2)
Test Method for Promoting Production Effect of Type IV Collagen and Type VII Collagen (1) Culture of human fibroblasts Human fibroblasts cultured in DMEM medium containing 10% FBS were seeded in a 24-well plate, and the cells were adhered. The medium was replaced with DMEM medium containing 25% FBS and 250 μM ascorbic acid glucoside, and the drug was added. One day later, the supernatant of the medium was collected and centrifuged, and the type IV and type VII collagens in the obtained supernatant were measured, and the amount of DNA was measured for the cells, which was used as an indicator of the number of cells.

(2) DNA quantification The amount of DNA was measured by a fluorescence measurement method using Hoechst H33342.

(3) Measurement of type IV and type VII collagen by sandwich ELISA
Type IV and type VII collagen were measured by sandwich ELISA. The antibodies used in this example are as follows.
Type IV collagen specific antibody; monoclonal antibody JK-199 and polyclonal antibody MO-S-CLIV
Type VII collagen specific antibody; monoclonal antibody NP-185 and monoclonal antibody NP-32

  When the amount of type IV and type VII collagen per DNA of the sample to which the drug is not added (control) is 100, the amount of type IV and type VII collagen per DNA of the drug added sample is expressed as type IV and type VII collagen. Production promotion rate. Table 2 below summarizes the collagen production promotion rates.

(Experiment Method-3)
Test Method for Promoting Type III Collagen Production (1) Culture of Human Skin Fibroblasts Human skin fibroblasts cultured in 10% FBS-containing DMEM medium were seeded in a 24-well plate, and the cells adhered, then 0.25 The medium was replaced with DMEM medium containing% FBS and 250 μM magnesium ascorbate phosphate, and the drug was added. After 3 days, the culture supernatant was collected and centrifuged. The amount of type III collagen and cellular DNA in the resulting supernatant was measured and used as an indicator of the number of cells.

(2) DNA quantification The amount of DNA was measured by a fluorescence measurement method using Hoechst H33342.

(3) Measurement of type III collagen The ability of cells to biosynthesize type III collagen is abbreviated as terminal peptide of type III collagen (Procollagen type III-peptide: PIIIP) secreted into the culture supernatant. ) It was evaluated by measuring the amount. Specifically, the measurement was performed using “Riagnost PIIIP (PIIIP) measurement kit (Chiide Diagnostics Co., Ltd.)”. The amount of type III collagen produced was divided by the amount of DNA and expressed as a value relative to a sample to which no plant extract was added (control) (see Table 3).

  From the above results, Dutch pepper extract was found to have a significant type III collagen production promoting effect.

(Experiment Method-4)
Method for evaluating the effect of human skin fibroblasts on type I collagen gel contractility After preparing a fibroblast (5 × 10 ⁴ cell / ml) suspension collagen solution (using type I collagen manufactured by Koken Co., Ltd.) on ice, Collagen was gelled at 37 ° C. Thereafter, a test substance (using DMSO as a control) and a DMEM medium containing 0.25% FBS were added, the gel was peeled from the petri dish wall surface, and collagen contraction was performed (each group n = 3). One or two days later, the diameter of the collagen gel was measured from three directions, and the average value was determined. The diameter before shrinkage was taken as the shrinkage rate of 0%, and the shrinkage rate after addition of the test substance was determined (see Table 4).

  From these results, the Dutch pepper extract was found to have a significant collagen gel contraction promoting action.

(Experiment Method-5)
Artificial skin production method Collagen gel is a gel solution of human dermis-derived fibroblasts (1 × 10 ⁵ cells / ml) suspended on ice and then gelled at 37 ° C. in a 60 mm Petri dish. did. Thereafter, the gel was peeled off from the petri dish wall surface, the collagen gel was placed on the metal, and a glass ring (inner diameter 12 mm) was placed on the gel. A KGM-DMEM (1: 1) mixed medium containing a human foreskin-derived epidermal keratinocyte suspension was added so as not to leak into the glass ring. Incubate overnight to allow epidermal cells to adhere and remove the ring the next day. A skin model having a multi-layered epidermis showing formation of the stratum corneum was prepared while filling the medium up to the boundary of the epidermal layer and exposing the epidermal layer to air.

  From day 4 after seeding of epidermal cells, (1) no drug added, (2) Dutch pepper extract 0.3%, (3) Dutch pepper extract 0.3% + mint extract 0.5%, (4) Pepper oil 0.3%, (5) Pepper oil 0.3% + Prunus extract 0.5%, (6) Pepper extract 0.3%, (7) Pepper extract 0.3% + Hypericum extract 0.5% (8) A medium containing 0.3% of dandelion extract, (9) 0.3% of dandelion extract + 0.5% of cypress extract, and then a medium containing the same kind and the same concentration every 2-3 days And further cultured for 2 weeks.

The formed artificial skin was stained with hematoxylin-eosin staining and immunostaining (anti-IV collagen antibody and anti-VII collagen antibody). The degree of staining of type IV and type VII collagen was scored from 5 to 1-5 in order from the lowest to the highest. Table 5 below summarizes the collagen staining scores.

  As described above, in the control (1), type IV collagen was weakly stained, but almost no type VII collagen was observed. On the other hand, it was confirmed in artificial skin that Dutch pepper, pepper, peanut, and fukidan popo, which have the action of promoting laminin-5 / IV / VII collagen production, both enhance the staining of IV / VII collagen. . Furthermore, it has been clarified that the serine protease inhibitors, mint, yew, hypericum and jujuak, further improve their dyeability.

(Experimental method-6)
The water content of the stratum corneum was measured when each of the extracts of Dutch pepper, pepper, sandalwood and dandelion was applied to the skin of three healthy men aged 30 to 45 years, and compared with the control site. Ion exchange water was used as a control. Each specimen was applied to a 2.5 cm square inside the forearm, and the water content of the stratum corneum immediately after application was measured using Corneometer CM825C (Courage + Khazaka) immediately after application to determine the water content increase rate. Table 6 below summarizes the rate of increase in the stratum corneum moisture when each plant extract is applied.

  From the above, it was shown that the amount of water in the stratum corneum was increased by applying the test substance, and there was a skin improvement effect.

(Experiment Method-7)
A continuous test of creams with the following prescription was conducted by a panel of healthy women aged 40-55 years. The 48 panelists were divided into 4 groups, and cream (invention product 1-4) was applied to half of the face of each group, and comparative product 1 was applied to the other half twice daily for 4 weeks. The stratum corneum moisture was measured with Corneometer CM825C, transdermal moisture transpiration was measured with Teweter TM210, and skin viscoelasticity was measured with a cutometer (both made by Courage + Khazaka). It was an index of degree. For the cute meter, the suction for 2 seconds and the release for 1 second are repeated three times in the usual way, and the first and third skin elongation (Uf value, maximum amplitude) and viscosity / elasticity ratio (Uv / Ue value) are calculated. Compared to before use. In addition, replicas of the corners of the eyes were collected with a silicone replica (Silflo, FLEXICO DEVELOPMENTS), the number and length of wrinkles were measured, and the values before and after continuous use were compared. Furthermore, from the degree of removal of the stratum corneum collected with the tape, the expert panel judged the visual feel of the skin texture and the uniform degree of removal of the stratum corneum.

  The results (compared values before continuous use) are shown in Tables 7 to 11 below. The numerical value in parentheses indicates the test value by Paired Student t-test.

  From the above results, the anti-wrinkle action of the product of the present invention was statistically significant.

  From the above results, the moisturizing effect of the product of the present invention was statistically significant.

  From the above results, the rough skin improvement effect of the product of the present invention was statistically significant.

以上の結果より、本発明品の皮膚弾力性増加が有意に確認された。

From the above results, it was confirmed that the skin elasticity of the product of the present invention was significantly increased.

  From the above results, the texture improvement effect of the product of the present invention was significantly recognized.

The compositions of the invention products 1 to 4 and the comparative product are shown in Table 12 below. In addition, a numerical value is represented by weight% with respect to the whole.

(Experimental method -8)
Anti-inflammation, wound healing promotion test Inflammation or wounds were artificially formed in 8-week-old HR-1 mice, (1) no drug added to each group (n = 5), (2) pepper oil 5%, ( 3) 0.5 g of 1,3-butylene glycol containing 5% peanut extract and (4) 5% poppy extract was applied twice a day for 5 days, and the condition of the inflamed or wounded site on the 5th day Was observed. The anti-inflammatory action and the wound healing promoting action were scored in 5 stages of 1-5 in order from the lowest to the highest. The results are summarized in Table 13 .

  As shown in Table 13, in the control (1), the degree of improvement of inflammation and wound was low. On the other hand, it has been confirmed in vivo that pepper, peanut and dandelion extract have an anti-inflammatory action and a wound healing promoting action.

(Example 1-Cream)
(1) Prescription (Phase A)
Stearic acid 10.0% by weight
Stearyl alcohol 4.0
Butyl stearate 8.0
Stearic acid monoglycerol ester 2.0
Vitamin E acetate 0.5
Vitamin A palmitate 0.1
Macadamia nut oil 1.0
Preservative Appropriate amount Perfume Appropriate amount (Phase B)
Glycerin 4.0
1,2-pentanediol 3.0
Potassium hydroxide 0.4
Magnesium ascorbate phosphate 0.1
L-arginine hydrochloride 0.01
Edetate trisodium 0.05
Dutch pepper 1,3-butylene glycol extract 0.5
Ion exchange water

(2) Manufacturing method The oil phase part of A and the aqueous phase part of B are each heated to 70 ° C. and completely dissolved. Add Phase A to Phase B and emulsify with an emulsifier. The emulsion is cooled using a heat exchanger.

(Example 2-Cream)
(1) Prescription stearic acid 5.0% by weight
Stearyl alcohol 4.0
Isopropyl myristate 18.0
Glycerol monostearate 3.0
Propylene glycol 10.0
Pepper methanol extract 0.01
Caustic potash 0.2
Sodium bisulfite 0.01
Preservative Appropriate amount of perfume Appropriate amount of ion-exchanged water

(2) Production method Propylene glycol, pepper methanol extract and caustic potash are added to ion-exchanged water, dissolved, heated and maintained at 70 ° C (aqueous phase). The other ingredients are mixed, dissolved by heating and kept at 70 ° C. (oil phase). The oil phase is gradually added to the aqueous phase, and after the addition is complete, the temperature is maintained for a while to cause the reaction. Thereafter, the mixture is uniformly emulsified with a homomixer and cooled to 30 ° C. while stirring well.

(Example 3-Wrinkle prevention cream)
(1) Prescription stearic acid 2.0% by weight
Stearyl alcohol 7.0
Hydrogenated lanolin 2.0
Squalane 5.0
2-Octyldodecyl alcohol 6.0
Polyoxyethylene (25 mol)
Cetyl alcohol ether 3.0
Glycerin monostearate ester 2.0
Propylene glycol 5.0
Peanut ethanol extract 0.05
Turmeric ethanol extract 0.05
Sodium bisulfite 0.03
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchanged water

(2) Manufacturing method Propylene glycol is added to ion-exchanged water and heated to maintain at 70 ° C. (aqueous phase). The other ingredients are mixed, dissolved by heating and kept at 70 ° C. (oil phase). The oil phase is added to the aqueous phase, pre-emulsified, and uniformly emulsified with a homomixer, and then cooled to 30 ° C. while stirring well.

Example 4 Anti-Aging Cream
(1) Prescription solid paraffin 5.0 wt%
Beeswax 10.0
Vaseline 15.0
Liquid paraffin 41.0
Glycerin monostearate ester 2.0
Polyoxyethylene (20 mol)
Sorbitan monolaurate 2.0
Soap powder 0.1
Borax 0.2
Fukitane popoacetone extract 0.05
Psychoethanol extract 0.05
Sodium bisulfite 0.03
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchanged water

(2) Manufacturing method Soap powder and borax are added to ion-exchanged water, heated and dissolved, and kept at 70 ° C. (aqueous phase). The other ingredients are mixed, heated and melted, and kept at 70 ° C. (oil phase). The reaction is gradually added while stirring the oil phase in the aqueous phase. After completion of the reaction, the mixture is uniformly emulsified with a homomixer and cooled to 30 ° C. while stirring well after emulsification.

Example 5-Emulsion
(1) Prescription stearic acid 1.0 wt%
Vaseline 5.0
Liquid paraffin 10.0
Polyoxyethylene (10 mol)
Monooleate 2.0
Polyethylene glycol 1500 3.0
Triethanolamine 1.0
Carboxyvinyl polymer 0.2
(Product name: Carbopol 941, BFGoodrich Chemical company)
Dutch mustard ethyl acetate extract 0.01
Sodium bisulfite 0.01
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchanged water

(2) Production method A carboxyvinyl polymer is dissolved in a small amount of ion-exchanged water (phase A). Polyethylene glycol 1500 and triethanolamine are added to the remaining ion-exchanged water, dissolved by heating and maintained at 70 ° C. (aqueous phase). The other ingredients are mixed, heated and melted, and kept at 70 ° C. (oil phase). Add the oil phase to the water phase, preliminarily emulsify, add the A phase and uniformly emulsify with a homomixer.

(Example 6-Emulsion)
(1) Prescription microcrystalline wax 1.0 wt%
Beeswax 2.0
Lanolin 2.0
Liquid paraffin 10.0
Squalane 5.0
Sorbitan sesquioleate 4.0
Polyoxyethylene (20 mol)
Sorbitan monooleate 1.0
Propylene glycol 7.0
Tomato 1,3-butylene glycol extract 10.0
Sodium bisulfite 0.01
Ethylparaben 0.3
Perfume Appropriate amount of ion-exchanged water

(2) Manufacturing method Propylene glycol is added to ion-exchanged water and heated to maintain at 70 ° C. (aqueous phase). The other ingredients are mixed, heated and melted and kept at 70 ° C. (oil phase). While stirring the oil phase, the aqueous phase is gradually added thereto and emulsified uniformly with a homomixer. Cool to 30 ° C. while stirring well after emulsification.

Example 7-Emulsion
(1) Prescription (Phase A)
Squalane 5.0 wt%
Oleolate 3.0
Vaseline 2.0
Sorbitan sesquioleate ester 0.8
Polyoxyethylene oleyl ether
(2OEO) 1.2
Evening primrose oil 0.5
Preservative Appropriate amount Perfume Appropriate amount (Phase B)
1,3-butylene glycol 4.5
Peanut ethanol extract 1.5
Ethanol 3.0
Carboxyvinyl polymer 0.2
Potassium hydroxide 0.1
L-Arginine L-Aspartate 0.01
Cascon ethanol extract 1.5
Erythritol 0.5
Sodium hexametaphosphate 0.05
Ion exchange water

(2) Manufacturing method The oil phase part of A and the aqueous phase part of B are each heated to 70 ° C. and completely dissolved. Add Phase A to Phase B and emulsify with an emulsifier. The emulsion is cooled using a heat exchanger.

Example 8-Jelly
(1) Formulation 95% ethyl alcohol 10.0% by weight
Dipropylene glycol 15.0
Polyoxyethylene (50 mol)
Oleyl alcohol ether 2.0
Carboxyvinyl polymer 1.0
(Product name: Carbopol 940, BFGoodrich Chemical company)
Caustic soda 0.15
L-Arginine 0.1
Fukitane popoethanol extract 7.0
2-hydroxy-4-methoxy sodium benzophenone sulfonate 0.05
Ethylenediamine tetraacetate
Trisodium 2 water 0.05
Methylparaben 0.2
Perfume Appropriate amount of ion-exchanged water

(2) Manufacturing method Carbopol 940 is uniformly dissolved in ion-exchanged water. On the other hand, Fukutan popoethanol extract and polyoxyethylene (50 mol) oleyl alcohol ether are dissolved in 95% ethanol and added to the aqueous phase. Next, after adding other components, the mixture is neutralized and thickened with caustic soda and L-arginine.

(Example 9-Cosmetic liquid)
(1) Prescription (Phase A)
Ethyl alcohol (95%) 10.0% by weight
Polyoxyethylene (20 mol)
Octyldodecanol 1.0
Pantotenyl ethyl ether 0.1
Dutch Pepper Methanol Extract 1.5
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
(Product name: Carbopol 940, BFGoodrich Chemical company)
Ion exchange water

(2) Manufacturing method A phase and C phase are each melt | dissolved uniformly, A phase is added to C phase, and it solubilizes. Next, filling is performed after adding phase B.

Example 10-Pack
(1) Prescription (Phase A)
Dipropylene glycol 5.0% by weight
Polyoxyethylene (60 mol)
Hardened castor oil 5.0
(Phase B)
Tomato extract 0.01
Olive oil 5.0
Tocopherol acetate 0.2
Ethylparaben 0.2
Fragrance 0.2
(Phase C)
Sodium bisulfite 0.03
Polyvinyl alcohol (degree of saponification 90, degree of polymerization 2,000) 13.0
Ethanol 7.0
Ion exchange water

(2) Manufacturing method A phase, B phase, and C phase are each uniformly dissolved, and B phase is added to A phase to solubilize. Next, this is added to phase C and then filled.

(Example 11-lotion)
(1) Prescription (Phase A)
Ethanol 5.0% by weight
POE oleyl alcohol ether 2.0
Oleyl alcohol 0.1
2-Ethylhexyl P-dimethylaminobenzoate 0.18
Perfume appropriate amount (phase B)
1,3-butylene glycol 9.5
Glycerin 2.0
Sodium pyrrolidonecarboxylate 0.5
Nicotinamide 0.3
Sandalwood 1,3-butylene glycol extract
0.1
β-cyclodextrin 1.0
Erythritol 0.05
Ion exchange water

(2) Production method The alcohol phase of A is added to the aqueous phase of B and solubilized to obtain a lotion.

Example 12-Solid Foundation
(1) Prescription talc 43.1% by weight
Kaolin 15.0
Sericite 10.0
Zinc flower 7.0
Titanium dioxide 3.8
Yellow iron oxide 2.9
Black iron oxide 0.2
Squalane 8.0
Isostearic acid 4.0
Monooleic acid POE sorbitan 3.0
Isocetyl octoate 2.0
Fukitan popo ethanol extract 0.5
Preservative Appropriate amount Fragrance Appropriate amount

(2) Manufacturing method The powder component of talc to black iron oxide is sufficiently mixed with a blender, and the oily component of squalane to isocetyl octoate, squirrhen popoethanol extract, preservative and fragrance are added and kneaded well, and then filled into a container. Mold.

(Example 13-emulsified foundation (cream type))
(1) Formula (powder part)
Titanium dioxide 10.3% by weight
Sericite 5.4
Kaolin 3.0
Yellow iron oxide 0.8
Bengala 0.3
Black iron oxide 0.2
(Oil phase)
Decamethylcyclopentasiloxane 11.5
Liquid paraffin 4.5
Polyoxyethylene-modified dimethylpolysiloxane 4.0
(Water phase)
Ion exchange water 50.0
1,3-butylene glycol 4.5
Dutch Pepper Ethanol Extract 1.5
Sorbitan sesquioleate 3.0
Preservative Appropriate amount Fragrance Appropriate amount

(2) Manufacturing method After heating and stirring the aqueous phase, a sufficiently mixed and pulverized powder part is added and homomixed. Furthermore, after adding the heat-mixed oil phase and carrying out a homomixer process, a fragrance | flavor is added with stirring and it cools to room temperature.

Claims (8)

  An anti-wrinkle agent comprising, as an active ingredient, any one or more of an extract derived from Dutch pepper, pepper, peanut and dandelion.   A basement membrane stabilizer comprising, as an active ingredient, any one or more of an extract derived from Dutch pepper, pepper, peanut and dandelion.   An extracellular matrix production promoter comprising as an active ingredient any one or more of an extract derived from Dutch pepper, pepper, peanut, and dandelion.   The extracellular matrix production according to claim 3, wherein the component of the extracellular matrix to be promoted is one or more selected from the group consisting of laminin 5, type III collagen, type IV collagen and type VII collagen. Accelerator.   The extracellular matrix production promoter according to claim 3 or 4, which promotes production of all of laminin 5, type III collagen, type IV collagen and type VII collagen.   An agent for improving skin sagging and firmness comprising, as an active ingredient, any one or more of an extract derived from Dutch pepper, pepper, peanut and dandelion.   A method for producing artificial skin, comprising a step of adding any one or more of extracts derived from Dutch pepper, pepper, sand bud and dandelion to an artificial skin formation medium.   A cosmetic method characterized by reducing wrinkles and improving sagging and firmness of skin by applying any one or more of an extract derived from Dutch pepper, pepper, peanut and dandelion to the skin.