CN112569162B

CN112569162B - Synergistic effect of radix Ophiopogonis extract, amino acid derivative and calcium lactate hydrate

Info

Publication number: CN112569162B
Application number: CN201910922080.9A
Authority: CN
Inventors: 安德鲁·菲利普·萨拉扎; 乔尔格·冯·哈根; 鲁楠; 亚历山德拉里·兰
Original assignee: Shanghai Baiqueling Biotechnology Co ltd; Merck Patent GmbH
Current assignee: Shanghai Baiqueling Biotechnology Co ltd; Merck Patent GmbH
Priority date: 2019-09-27
Filing date: 2019-09-27
Publication date: 2024-04-12
Anticipated expiration: 2039-09-27
Also published as: CN112569162A

Abstract

The present invention relates to an active mixture consisting of component a), component b), component c) and component d), said component a) being an herbal extract obtained from ophiopogon root (Ophiopogon japonicus), said component b) being a mixture of serine, arginine and histidine, said component c) being pyrrolidone-5-carboxylic acid, and said component d) being calcium lactate hydrate, and its use in cosmetic and/or dermatological compositions. The invention also relates to a cosmetic and/or dermatological composition comprising: a) Herbal extract obtained from ophiopogon root (Ophiopogon japonicus); b) Amino acid: serine, arginine, and histidine; c) Amino acid derivative: pyrrolidone-5-carboxylic acid; and d) calcium lactate hydrate. The invention also relates to a non-therapeutic method by topical application of said active mixture or of said cosmetic or dermatological composition.

Description

Synergistic effect of radix Ophiopogonis extract, amino acid derivative and calcium lactate hydrate

Technical Field

The present invention relates to the field of cosmetics, and also to an active mixture consisting of component a), component b), component c) and component d), said component a) being an herbal extract obtained from ophiopogon root (Ophiopogon japonicus), said component b) being a mixture of serine, arginine and histidine, said component c) being pyrrolidone-5-carboxylic acid, said component d) being calcium lactate hydrate, and its use in cosmetic and/or dermatological compositions. The invention also relates to a cosmetic and/or dermatological composition comprising:

a) Herbal extract obtained from ophiopogon root (Ophiopogon japonicus);

b) Amino acid: serine, arginine, and histidine;

c) Amino acid derivative: pyrrolidone-5-carboxylic acid; and

d) Calcium lactate hydrate.

The invention also relates to a non-therapeutic method by topical application of said active mixture or of said cosmetic or dermatological composition.

Background

The epidermis is the outermost layer of the skin and is mainly composed of keratinocytes (keratinocytes). These cells differentiate upward to form the basement membrane (basal layer) where they contact the dermis to form layered layers (e.g., the stratum spinosum and stratum granulosum), ultimately as sheets of connective keratinocytes located at the outermost location, the Stratum Corneum (SC). Each stage of epidermal differentiation is characterized by the expression of a specific protein.

ChunSik Choe et al, nature Scientific Reports, volume 7, 15900 (2017), pages 1-13 describe that SCs are considered skin barriers that prevent penetration of exogenous substances into the skin and also prevent evaporation of water away from the body. The barrier function of SC is mainly maintained by the lateral packing order of intercellular lipids. Furthermore, a three-layer model of keratin-water interactions in SC is explained. At the uppermost layer (30-0% sc depth), the keratin chains are highly folded and remain in the most stable alpha-helical conformational form. Therefore, water cannot be embedded in the keratin chain and there are no vacancies to bind water molecules to the keratin. It was further found that natural moisturizing factor NMF is mainly responsible for binding water in these SC layers. At the middle layer (70-30% sc depth), the keratin chains spread more and therefore the interactions between keratin side chains are weak, and water molecules intercalate into the keratin filaments and bind there, causing the keratinocytes (corneocells) to swell. At the bottom layer (100-80% sc depth), the keratin chains have fewer vacancy binding sites than the middle layer, since keratin folding is larger than at 70% sc depth, with the lowest NMF concentration and maximum amount of water. Thus, in this SC layer, the incoming water is not able to effectively form hydrogen bonds with the surrounding keratin and NMF, which makes the SC layer the bottommost low expansion zone.

Aged and unhealthy skin is characterized by dryness and reduced barrier function. If not addressed in time, these physical attributes may cause further adverse effects on the skin, including age-related chronic inflammation and irritation, leading to further tissue damage. This results in a significant change in the composition and functional capacity of the epidermis.

In order to prevent diseases associated with dry skin, impaired barrier function or redness, traditional approaches have focused on "hydration" of the SC. However, the biochemical processes associated with silk fibroin also appear to play an important role.

Aileen Sandilands et al, "Filaggrin in the frontline: role in skin barrier function and disease ", journal of Cell Science122 (2009), 1285-1294 describe that the absence of profilaggrin or filaggrin results in the formation of a poor stratum corneum, which is prone to water loss. Further, recent human genetics studies strongly suggest that disturbance of skin barrier function leads to enhanced transdermal transfer of allergens due to reduced or complete loss of silk fibroin expression.

Natural Moisturizing Factors (NMF) are the products of silk fibroin degradation in SCs and SG. The modified silk polyprotein results in a reduction of NMF in the Stratum Corneum (SC) and the Stratum Granulosum (SG).

Nobutomo Ikarashi et al, "Relationship between Aging-related skin dryness and aquaporins", international Journal of Molecular Sciences,2017,18,1559 describe a study which analyzed the mechanism of aging-related skin dryness, focusing on aquaporin, a water channel. The research results show that the expression level of aquaporin in skin decreases with aging, which suggests that the aquaporin may become a new target for preventing and treating aging-related skin dryness.

Thus, methods of skin dryness and improved barrier function can be addressed by stimulating skin cells to synthesize macromolecules involved in these processes, such as silk fibroin or aquaporins.

Hyaluronic acid is the main water-binding molecule in the dermis. This function is also performed in the epidermis together with keratin, silk fibroin and natural moisturizing factors.

Radix Ophiopogonis (also called Dwarf liytiturf, radix Ophiopogonis, fountaInplant, monkeygrass, leptoradix Serpentis) is a species of Ophiopogon native to China, india, japan and Vietnam. It is a perennial plant that forms turf. The leaves are linear and 20-40 cm long. The flowers were pale white pale purple. In the ground, this species has large stolons and tubers.

In traditional Chinese medicine, radix Ophiopogonis tuberous root is called radix Ophiopogonis, which is the main herb for yin deficiency. According to the Chinese herbal medicine (Chinese Herbal Medicine Materia Medica), the Chinese herbal medicine is sweet, slightly bitter and slightly cold, enters heart, lung and stomach channels, nourishes yin and stomach, spleen, heart and lung, clears heat, calms and removes agitation, moistens intestinal tracts, strengthens the functions of stomach and nourishes lung. Some of the major components of this herb are reported to be stigmasterols, mucilages, saponins, carotenoids, steroid saponins and polysaccharides.

Polysaccharide is described in Pengjiao Zeng et al Progress in Molecular Biology and Translational Science,2019,3.6"Ophiopogon japonicus" as the main component of ophiopogon root, with an extraction yield of up to 35%. The molecular weight of ophiopogon polysaccharides is not described consistently, ranging from 2.74 to 325kDa, consisting mainly of beta-fructose and small amounts of alpha-glucose. Levan is a polymer of fructose molecules.

US20100303872 describes a plant extract enriched in polyphenols, wherein the plant extract is obtained from rose in combination with at least one hydrating agent or humectant.

US20110045105 describes the use of ophiopogon japonicus extract as active ingredient, which preferably acts by increasing the level of NMF of the stratum corneum, in particular by increasing the level of lactate and serine. It further describes the cosmetic use of ophiopogon japonicus extract to improve and/or enhance the barrier function of the skin by increasing the formation of tight junctions, in particular by stimulating the synthesis of tight junction protein-1 and zonula occludens ZO-1.

US20190060390 describes a method of treating atopic dermatitis in a human subject or animal, the method comprising topically administering to the human subject or animal an active ingredient obtained from dwarf lilyturf tuber.

Amino acid chelated minerals are known from WO2007/084932 as nutritional supplements. The reported mixed amino acid/mineral compounds include minerals that bind to a first amino acid that is different from a second amino acid and are characterized by increased solubility and/or increased absorption through the Gastrointestinal (GI) tract as compared to compounds having only the first amino acid or the second amino acid.

US5,516,925 describes amino acid chelated mineral compositions containing amino acid ligands with improved palatability.

US2008/0260672 describes an aqueous preservative solution having a high amino acid content, which has a special preservative effect against fungi, which has excellent storage stability, and which is capable of imparting a high moisturizing effect without causing a sticky feeling when incorporated into cosmetics, and has an effect of preventing dyed hair from fading. Such compositions comprise pyrrolidone carboxylic acid and/or salts thereof, basic amino acids and/or salts thereof, acidic amino acids and/or salts thereof, neutral amino acids and/or salts thereof and lactic acid and/or salts thereof, incorporated in specific proportions and having specific pH.

WO2008/142147 describes a composition comprising pyrrolidone-5-carboxylic acid and at least one compound from the racemic or isomeric forms of citrulline, arginine and asparagine, and salts thereof, in a physiologically acceptable carrier for the preparation of a medicament for use in the treatment and/or prevention of atopic dermatitis.

KR1020090054777 describes a composition comprising lactic acid or a salt thereof, pyrrolidone carboxylic acid or a salt thereof and an amino acid for controlling the pH of skin, and a cosmetic composition comprising the same. Such compositions have also been reported to protect skin barrier function, and the components of the composition are part of the Natural Moisturizing Factor (NMF). As a result, the composition or cosmetic containing the composition is reported to be capable of moisturizing skin.

EP2404502 describes compositions containing pyrrolidone carboxylic acid and metal salts, such as copper, zinc or manganese salts, which synergistically enhance the antiviral and antibacterial effects of PCA.

WO2017/042748 describes a moisturizing cosmetic composition comprising at least one cosmetically acceptable lactate salt, at least one amino acid selected from the group consisting of L-serine, glycine, leucine, lysine, proline, valine, tyrosine, glutamic acid, alanine, aspartic acid, arginine, tryptophan, histidine, phenylalanine, ornithine, threonine and mixtures thereof, trehalose and at least one cosmetically acceptable zinc salt.

However, there remains a need to find a powerful solution to improve skin dryness associated with aging and to reduce or prevent chronic inflammation (inflammation) and irritation associated with aging.

It is therefore an object of the present application to provide such a solution.

Summary of The Invention

The inventors have now found that the above object can be achieved by the methods described herein or by specific mixtures of the present application.

The invention relates to an active mixture consisting of:

a) Herbal extract obtained from ophiopogon root (Ophiopogon japonicus);

b) Amino acid: serine, arginine, and histidine;

c) Amino acid derivative: pyrrolidone-5-carboxylic acid; and

d) Calcium lactate hydrate.

The invention also relates to the use of the active mixture for producing a cosmetic and/or dermatological composition.

The invention also relates to a cosmetic and/or dermatological composition comprising:

a) Herbal extract obtained from ophiopogon root (Ophiopogon japonicus);

b) Amino acid: serine, arginine, and histidine;

c) Amino acid derivative: pyrrolidone-5-carboxylic acid; and

d) Calcium lactate hydrate.

The invention also relates to non-therapeutic methods for improving age-related skin dryness or for reducing or preventing age-related chronic inflammation and irritation by topically applying an active mixture or a cosmetic composition comprising said components of said mixture.

The invention also relates to a non-therapeutic method for stimulating the synthesis of hyaluronic acid, aquaporins and filaggrin in living cells of the epidermis and dermis by topical application of an active mixture or of a cosmetic composition comprising said components of said mixture.

Drawings

FIG. 1 shows the analysis of gene expression profile of HAS1 as described in example 1.

FIG. 2 shows the analysis of the gene expression profile of aquaporin 1 as described in example 2.

FIG. 3 shows immunofluorescent staining of epidermal barrier protein as described in example 3.

FIG. 4 shows quantitative measurements of silk fibroin as described in example 4.

Detailed Description

"active mixture" in the sense of the present invention means a mixture of all components a), b), c) and d) which have a synergistic effect on skin cells, preferably on human skin cells.

As previously mentioned, preferably the said member

a) Herbal extract obtained from ophiopogon root (Ophiopogon japonicus);

b) Amino acid: serine, arginine, and histidine;

c) Amino acid derivative: pyrrolidone-5-carboxylic acid; and

d) The active mixture of calcium lactate hydrate is characterized in that component a) is an herbal extract obtained from the root of dwarf lilyturf tuber (Ophiopogon japonicus).

Particularly preferably, the active mixtures according to the invention are characterized in that component a) contains an enzymatic hydrolysate of radix Ophiopogonis roots.

Preferably, the ophiopogon root extract can be obtained by a process comprising the following stages:

dissolving radix Ophiopogonis root tuber powder in water,

at least one enzymatic hydrolysis step carried out,

-separating the soluble and insoluble phases, and

enzymatic inactivation by heat treatment.

These stages are common in the field of plant extraction and the person skilled in the art is able to adjust their reaction parameters based on their general knowledge.

The enzymatic inactivation may be followed by one or more filtration and/or concentration stages. Preferably, the enzymatic inactivation is followed by at least one filtration stage and one sterile filtration stage. The enzyme used is a carbohydrate enzyme. They may be purified enzymes, single component enzymes or mixtures of different enzymes.

Preferably, the ophiopogon japonicus extract obtained by said method comprises at least 60% by weight of levan, preferably at least 80%, relative to the total weight of dry matter. Preferably, the fructose content of the material is 28 to 245g/l, preferably 85 to 127g/l. Fructose was metered spectrophotometrically according to the method for determining ketose described in boratenski, j., analytical Biochemistry,1984,137,2,528-532.

Preferred aqueous extracts of radix Ophiopogonis have 95 to 99% carbohydrates (including polysaccharides) and 1 to 5% polyphenols.

Particularly preferably, the active mixture according to the invention is characterized in that component a) is an extract of radix Ophiopogonis root, which comprises water, 1, 2-hexanediol and octanediol.

Component a) is particularly preferably an aqueous solution composed of 85.90% to 90.10% of water, 9.00% to 13.00% of the extract of ophiopogon root as described above, 0.45% to 0.55% of 1, 2-hexanediol and 0.45% to 0.55% of octanediol, under the trade nameAre marketed by the company Silab, france.

The invention also relates to the use of said active mixtures as described above or of active mixtures having the preferred description of components a), b), c) and d) as described hereinbefore or hereinafter for the preparation of cosmetic and/or dermatological compositions.

The cosmetic and/or dermatological composition may comprise an active mixture as described previously or preferably as described previously in an amount of from 0.1 to 20% by weight, preferably from 0.5 to 10% by weight, more preferably from 1 to 6% by weight, based on the total weight of the cosmetic and/or dermatological composition.

The invention further relates to a cosmetic and/or dermatological composition comprising components a), b), c) and d) as described before or preferably before or after it.

In the cosmetic and/or dermatological composition, the total weight content of components a), b), c) and d) is from 0.1 to 20% by weight, preferably from 0.5 to 10% by weight, more preferably from 1 to 6% by weight. Based on the total weight of the cosmetic and/or dermatological composition.

The amino acids, amino acid derivatives and calcium lactate hydrate in the active mixture and/or composition include all stereoisomers or racemic mixtures.

L-serine is preferably used. L-arginine is preferably used. L-histidine is preferably used. L-pyrrolidone-5-carboxylic acid is preferably used.

Preferably, L-calcium lactate hydrate is used. The preferred L-calcium lactate hydrate is L-calcium lactate pentahydrate.

In the mixture or cosmetic or dermatological composition as described above, 0.2 to 0.5 equivalent of calcium lactate hydrate should preferably be used together with 1 equivalent of serine. When used according to the invention, it is particularly preferred that 0.3 to 0.4 equivalent of calcium lactate hydrate is used together with 1 equivalent of serine. When used according to the invention, it is particularly preferred to use 1:0.33 of serine and calcium lactate hydrate.

Preferably the amount of serine is the highest amount in the combination of components b), c) and d) in the active mixture or the cosmetic or dermatological composition according to the invention.

Preferably, the amount of pyrrolidone-5-carboxylic acid, in particular L-pyrrolidone-5-carboxylic acid, is the second highest amount in the combination of components b), c) and d) in the active mixture or the cosmetic or dermatological composition according to the invention.

Preferably, L-serine, L-arginine, L-histidine, L-pyrrolidone-5-carboxylic acid and L-calcium lactate pentahydrate are used in the active mixtures or cosmetic or dermatological compositions of the invention.

In components b), c) and d) of the active mixture, the serine content is preferably 25 to 35% by weight, the arginine content is preferably 5 to 10% by weight, and the histidine content is preferably 5 to 10% by weight. Preferably from 10 to 30% by weight of pyrrolidone-5-carboxylic acid, preferably from 25 to 35% by weight of calcium lactate hydrate, wherein the combined total of components b), c) and d) amounts to 100% by weight.

The content of L-serine in components b), c) and d) in the active mixture is preferably 25 to 35% by weight, the content of L-arginine is preferably 5 to 10% by weight, the content of L-histidine is preferably 5 to 10% by weight, the content of L-pyrrolidone-5-carboxylic acid is preferably 10 to 30% by weight, and the content of L-calcium lactate pentahydrate is preferably 25 to 35% by weight, where the combined total of components b), c) and d) is 100% by weight.

The following amounts of components are particularly preferred: 29 to 31% by weight of L-serine, 7 to 8% by weight of L-arginine, 7 to 8% by weight of L-histidine, 20 to 25% by weight of L-pyrrolidone-5-carboxylic acid and 29 to 31% by weight of L-calcium lactate pentahydrate, wherein the combined total of components b), c) and d) is 100% by weight.

Preferably, a mixture consisting of 30.6% by weight of L-serine, 7.5% by weight of L-arginine, 7.3% by weight of L-histidine, 24.2% by weight of L-pyrrolidone-5-carboxylic acid and 30.4% by weight of L-calcium lactate pentahydrate is combined with component a) as described or preferably as described above to construct a preferred active mixture according to the invention.

The active mixtures as described previously or preferably as described above preferably contain the sum of component a) and components b), c) and d) in a ratio of 5:1 to 1:5, preferably in a ratio of 3:1 to 1:3, particularly preferably in a weight ratio of 1:1,3:1 and 1:3.

cosmetic and/or dermatological compositions as described previously or preferably as described previously preferably contain the sum of component a) and component b), c) and d), in a ratio of 5:1 to 1:5, preferably in a ratio of 3:1 to 1:3, particularly preferably in a weight ratio of 1:1,3:1 and 1:3.

for the purposes of the present invention, the term "composition" is also used synonymously with the term "formulation".

The formulations herein are typically cosmetic or dermatological formulations, which may be applied topically. By "topically applicable" it is meant that the formulation is applied externally and topically, i.e. the formulation must be suitable, e.g. capable of being applied to the skin. In this case, the formulation comprises a cosmetically, pharmaceutically or dermatologically suitable carrier, and optionally further comprises other suitable ingredients, depending on the desired property profile. Topical formulations are preferably used as cosmetic compositions. Suitable carriers and adjuvants or fillers are known in the art.

The formulations according to the invention may also contain abrasives, anti-acne agents, anti-skin ageing agents, anti-cellulite agents, anti-dandruff agents, anti-inflammatory agents, irritation preventative agents, irritation inhibitors, antioxidants, astringents, sweat inhibitors, preservatives, antistatic agents, adhesives, buffers, carrier materials, chelating agents, cell stimulating agents, cleaning agents, care agents, depilatories, surfactants, deodorants, antiperspirants, softening agents, emulsifiers, enzymes, essential oils, fibers, film formers, fixatives, foam formers, foam stabilizers, substances to prevent foaming, foam promoters, gelling agents, gel formers, hair care agents, hair styling agents, hair straightening agents, moisturizers, humectants, moisturizing substances, bleaching agents, reinforcing agents, stain removers, optical brighteners, impregnating agents, anti-fouling agents, friction reducers, lubricants, sunscreens, plasticizers, coating agents, polishing agents, gloss agents, polymers, proteins, re-lubricants, abrasives, silicones, skin soothing agents, skin cleansers, skin care agents, skin restoration agents, skin lightening agents, skin care agents, skin softeners, hair promoters, cooling agents, skin cooling agents, warming agents, skin warming agents, stabilizers, ultraviolet light absorbers, organic ultraviolet filters, inorganic ultraviolet filters, detergents, fabric conditioners, suspending agents, skin tanning agents, thickeners, vitamin oils, waxes, fats, phospholipids, saturated fatty acids, mono-or polyunsaturated fatty acids, alpha-hydroxy acids, polyhydroxy fatty acids, liquefying agents, dyes, color protectants, pigments, preservatives, fragrances, flavoring substances, odoriferous substances, polyols, surfactants or electrolytes.

Particularly suitable active ingredients for the combination are escitalopram @Ectoin), hydroxyescin, trehalose, glycerol, glycosylglycerol, beta-mannosylglycerin (firoin), beta-mannosylglyceramide (firoin A), inositol di-phosphate (DIP), cyclic 2, 3-diphosphoglycerate (cDPG), 1-diglycerol phosphate (DGP), inositol di-mannosyl Diphosphate (DMIP), betaine, glycine betaine, proline betaine, glutamic acid betaine, alanine, proline, glutamine, N-acetyllysine, glutamine 1-amide, taurine, choline O-sulfate, carnitine, arsenic betaine, crotonobetaine, dimethyl sulfoacetate, dimethyl sulfonate, homobetaine, trimethylamine N-oxide, panthenol, sorbitol, meglumine, hyaluronic acid or hyaluronic acid derivatives, urea>Urea) and nicotinamideNicotinamide), 5, 7-dihydroxy-2-methylchromone under the trade name +.>Marketing or commercial products-> RenouMer，/> VTA， Poppy SE，/>Isoque Ricin or +.> Cyclopeptide 5，/>Cyclopeptide 5 is alcohol-free, < - > and-> SereneShield，Emblica，Or->A liquid.

The formulation may comprise or consist essentially of or consist of the necessary or optional components mentioned above and/or below. All compounds or components useful in the formulation are known and commercially available or can be synthesized by known methods.

These further active ingredients are preferably present in the topical formulation in an amount of from 0.01 to 20% by weight, particularly preferably from 0.1 to 15% by weight, very particularly preferably from 0.2 to 8% by weight, based on the total amount of the formulation.

Organic uv filters, so-called hydrophilic or lipophilic sunscreen filters, are effective (absorbers) in the UVA region and/or UVB region and/or IR and/or VIS region. These substances may be chosen in particular from dibenzoylmethane derivatives, cinnamic acid derivatives, salicylic acid derivatives, camphor derivatives, triazine derivatives, beta-diphenylacrylate derivatives, para-aminobenzoic acid derivatives and polymeric filters and silicone filters. The UV filters are generally named according to INCI nomenclature as follows.

Particularly suitable for combination with the active mixtures of the invention are ethylhexyl salicylate, phenylbenzimidazole sulfonic acid, benzophenone-3, benzophenone-4, benzophenone-5, 2- (4-diethylamino-2-hydroxybenzoyl) n-hexyl benzoate, 4-methylbenzylidene camphor, terephthaloyl dicarbamate sulfonic acid, disodium phenyldibenzimidazole tetrasulfonate, methylenebis (benzotriazolyl) tetramethylbutylphenol, butylmethoxydibenzoylmethane, ethylhexyl triazone, diethylhexyl butyramide triazone, triazolo cresol trisiloxane, phenylene bis-diphenyltriazine, polysiloxane-15,1,1-dicarboxy (2, 2' -dimethylpropyl) -4, 4-diphenylbutadiene, 2, 4-bis [5-1 (dimethylpropyl) benzoxazol-2-yl (4-phenyl) imino ] -6- (2-ethylhexyl) imino-1, 3, 5-triazine, and mixtures thereof. These organic UV filters are generally incorporated into the formulation in an amount of from 0.01 to 20% by weight, preferably from 1 to 10% by weight.

The formulations may preferably comprise adjuvants such as cosmetic oils (e.g. caprylic/capric triglyceride, C12-15 alkyl benzoate, isopropyl myristate, arylalkyl benzoate, e.g. phenethyl benzoate (X-Tend 226) or cosmol brand oil components, such as dimyristol tartrate, tri-C14-C15 alkyl citrate, C12-C13 alkyl lactate, tridecyl salicylate, C12-C13 alkyl caprylate, C12-C13 alkyl malate, C12-C13 alkyl citrate, C12-C13 alkyl tartrate, or polar-protic adjuvants (e.g. propylene glycol, glycerol, isopropanol, ethanol) or so-called solubilisers (e.g. butylphthalimide, isopropylphthalimide, dimethyl isosorbide) very particularly preferred cosmetic oils are C12-C13 alkyl lactate (commercially available under the trade name cosmol ELI) and phenylacetate (commercially available under the trade name X-Tend 226).

Formulations as described above can be synthesized by mixing the active mixtures or components a), b), c) and d) according to the invention with carriers suitable for such formulations and optionally with auxiliaries and/or fillers. Suitable carriers and adjuvants or fillers are described in detail in the following sections.

The components of the formulation may be incorporated in a conventional manner by means of techniques well known to those skilled in the art.

Formulations suitable for external use may be applied or sprayed onto the skin, for example, as creams or milks (O/W, W/O, O/W/O/W, W/Si), as lotions or emulsions, in the form of oil-alcohol, oil-water or water-alcohol gels or solutions. They may be in the form of solid sticks or formulated as aerosols. They may be shampoos, body washes, facial cleansers or essences.

Hereinafter, for example, application forms as formulations to be used may be mentioned: solutions, suspensions, emulsions, PIT emulsions, pastes, ointments, gels, creams, lotions, powders, oils, aerosols, plasters, pressed cloths, bandages and sprays.

Preferred adjuvants are from the group of preservatives, stabilizers, solubilizers, colorants, odor improvers, thickeners, plasticizers, humectants, surfactants, emulsifiers, preservatives, defoamers, fragrances, waxes, lanolin, propellants and other ingredients customary in cosmetics.

Ointments, pastes, creams and gels may contain conventional carriers suitable for topical application, for example animal and vegetable fats, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silica, talc and zinc oxide, or mixtures of these substances.

Powders and sprays can contain customary carriers, for example lactose, talc, silica, aluminum hydroxide, calcium silicate and polyamide powders or mixtures of these substances. The spray may additionally comprise conventional volatile liquefied propellants such as chlorofluorohydrocarbons, propane/butane or dimethyl ether. Compressed air may also be used advantageously.

The solutions and emulsions may contain conventional carriers such as solvents, solubilisers and emulsifiers such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylene glycol, oils, in particular cottonseed oil, groundnut oil, wheat germ oil, olive oil, castor oil and sesame oil, XTend 226, glycerol fatty acid esters, polyethylene glycol and fatty acid esters of sorbitan, or mixtures of these substances.

Suspensions may include conventional carriers, such as liquid diluents, for example water, ethanol or propylene glycol, suspending agents, such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and polyoxyethylene sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances.

The facial and body oils may comprise conventional carriers such as synthetic oils, for example fatty acid esters, fatty alcohols, silicone oils, natural oils, for example vegetable oils and oily plant extracts, paraffinic oils, lanolin oils or mixtures of these substances.

Further typical cosmetic applications are lipsticks, lip care sticks, powder cosmetics, emulsion cosmetics and wax cosmetics, as well as sunscreens, pre-sun and post-sun preparations.

Preferred formulations also include, inter alia, emulsions.

Emulsions are advantageous and include, for example, the fats, oils, waxes and other fatty substances, as well as water and emulsifiers, as are commonly used in such formulations. Emulsifiers which can be used are, for example, the known W/O and O/W emulsifiers. It is advantageous to use other conventional co-emulsifiers in the preferred O/W emulsions.

Preferred oils and/or lipids for the topical formulation include: paraffin, isoparaffin, dioctyl ether, PPG-15, stearyl ether, beeswax, candelilla wax, carnauba wax, ethylhexyl stearate, caprylic/capric triglyceride, cetyl lactate, stearic stearate, isononyl isononanoate, octyldodecanol, hexyldecanol, squalene, natural triglycerides such as cherry seed oil (trunk Cerasus), avocado oil, safflower seed oil, macadamia seed oil, cocoa butter (theoberma Cacao), butter oil and mixtures thereof.

Preferred absorption and/or thickening agents for the formulation include modified corn starch, silica, talc, zinc stearate, magnesium sulfate, zinc oxide, calcium and aluminum borosilicate, starch and derivatives, polyurethane, and mixtures thereof.

As previously described or preferably as previously described, the active mixture synergistically stimulates the synthesis of hyaluronic acid by HAS1, synergistically stimulates the synthesis of the water transporter AQP1 and synergistically increases the filaggrin content in living cells of the epidermis and dermis.

By this unique combination of properties, the active mixture is able to prevent dryness associated with skin aging and synergistically enhance the barrier function of the epidermis.

The invention also relates to a non-therapeutic method for improving skin dryness associated with aging by topically applying to human skin an active mixture as described above or by topically applying a cosmetic or dermatological composition comprising an active mixture as described above or a cosmetic or dermatological composition comprising components a), b), c) and d) as described above. Preferably, the non-therapeutic method is a cosmetic method of applying a cosmetic composition.

The invention also relates to a non-therapeutic method for reducing or preventing chronic inflammation and irritation associated with aging by topically applying to human skin an active mixture as described above or by topically applying a cosmetic or dermatological composition comprising an active mixture as described above or a cosmetic or dermatological composition comprising components a), b), c) and d) as described above. Preferably, the non-therapeutic method is a cosmetic method of applying a cosmetic composition.

In a further aspect, the invention also relates to a non-therapeutic method for stimulating the synthesis of hyaluronic acid, aquaporins and filaggrin in living cells of the epidermis and dermis by topical application to human skin of an active mixture as described above or by topical application of a cosmetic or dermatological composition comprising an active mixture as described above or of a cosmetic or dermatological composition comprising components a), b), c) and d) as described above. Preferably, the non-therapeutic method is a cosmetic method of applying a cosmetic composition.

The application is carried out using standard techniques, for example by applying shampoos, body washes, creams, face washes, pastes, gels, lotions, essences to the skin to be treated, or dissolving a predetermined amount of a formulation comprising an active mixture as described above or an active mixture as described above in water and subsequently using the foam mixed or formed from said water for cleansing or skin treatment.

It should be noted that variations of the embodiments described in the present invention are included in the scope of the present invention. Any feature disclosed in this application may be replaced by alternative features serving the same, equivalent or similar purpose, unless expressly excluded. Thus, unless otherwise indicated, any feature disclosed in this specification should be considered as an example of a generic series or equivalent or similar feature.

All of the features of the invention may be combined with each other in any way unless the specific features and/or steps are mutually exclusive. This is especially true for the preferred features of the invention. Also, features that are not necessarily combined may be used alone (rather than in combination).

The technical teachings disclosed in the present invention may be abstracted and combined with other examples.

The complete disclosure of all applications and publications mentioned above and below are incorporated by reference into the present application.

The following examples are intended to illustrate the invention. However, they should not be considered limiting in any way.

Materials:

the components b), c) and d) used in this example consist of the following ingredients disclosed in Table 1. The mixture is hereinafter referred to as mixture 1.

Table 1:

pca=pyrrolidone-5-carboxylic acid

Component a) is a productSP, as described in table 2, will be named OJ hereinafter. It is used at a concentration of 1%.

Table 2:

the active mixture used in the examples is OJ to mixture 1 in a weight ratio of 2: 1. The mixture is hereinafter referred to as oj+nmf.

Example 1:

human epidermal keratinocyte cell line HaCAaT cells suitable for growth in DMEM (Thermo Fisher, art, germany) supplemented with 10% fcs (Biochrom, merck, germany) were seeded onto coverslips in 6-well plates at 37 ℃,5% co ₂ At 0.5×10 ⁵ Individual cells/cm ² Is cultured at a density of 24 hours. Thereafter, the cells were treated with OJ (1% v/v) or OJ+NMF (1% v/v+0.5% w/v) for 48 hours and incubated under the same conditions.

In this experiment, it was analyzed whether OJ and oj+nmf as active mixtures could increase proteins that play a role in the synthesis of macromolecules involved in skin moisture retention. The key molecule in skin hydration is hyaluronic acid. The molecule is synthesized by hyaluronate synthase 1 (HAS-1) and hyaluronate synthase 2 (HAS-2).

Gene expression analysis was performed using primers from a universal probe pool in the form of a custom assay supplied by the RealTime Ready System (Roche, manheim, germany). For this purpose, RNA was isolated with the aid of the RNeasy Mini Kit (QIAGEN, hilden, germany). cDNA was synthesized from the isolated RNA using the High-Capacity cDNA Reverse Transcription kit with RNase inhibitor kit (Thermo, darmstadt, germany). Finally, LC480 (Roche, manheim, germany) was used to analyze gene expression according to the cycling conditions recommended by the manufacturer. Use 2 ^-ΔΔCt The method calculates fold change from control.

Results 1:

as shown in FIG. 1, untreated HaCaT cells did produce basal levels of HAS-1, which was set to 1 as a reference. HAS-1RNA was up-regulated by 58% with OJ treatment, whereas the active mix of OJ and mix 1 (OJ+NMF) up-regulated HAS-1RNA by 258%. UT is an untreated control.

Example 2:

human epidermal keratinocyte cell line HaCAaT cells suitable for growth in DMEM (Thermo Fisher, germany Art.11995123) supplemented with 10% FCS (Biochrom, merck, germany) were seeded onto coverslips in 6-well plates, in37℃，5％CO ₂ At 0.5×10 ⁵ Individual cells/cm ² Is cultured at a density of 24 hours. Thereafter, the cells were treated with mixture 1 (0, 5% w/v), OJ (1% v/v) or OJ+NMF (1% v/v+0.5% w/v) for 48 hours and incubated under the same conditions.

Here we analyzed the effect on the storage of water in the cells and the fluidity of water in the different layers of the cells (regulated by aquaporins) and quantified the expression profile of the aquaporin family.

As shown in fig. 2, an upregulation of 161% of aquaporin 1 was detected by treatment with OJ, and the active mix of oj+nmf increased expression to 233%. UT is untreated control. No upregulation by mixture 1 was detected.

Example 3:

human epidermal keratinocyte cell line HaCAaT cells suitable for growth in DMEM (Thermo Fisher, art, germany) supplemented with 10% fcs (Biochrom, merck, germany) were seeded onto coverslips in 6-well plates at 37 ℃,5% co ₂ At 0.5×10 ⁵ Individual cells/cm ² Is cultured at a density of 24 hours. Thereafter, the cells were treated with mixture 1 (0.5%), OJ (1%) or OJ+NMF (1% v/v+0.5% w/v) for 48 hours and incubated under the same conditions.

This experiment was performed to analyze the effect on the expression of skin barrier proteins tight junction proteins, closed bands and filaggrin after 48 hours of treatment with the active mixture oj+nmf. The increase in tight junctions and closed bands indicates improved binding of keratinocytes to each other. Whereas, since degradation of silk fibroin produces natural moisturizing factors, an increase in silk fibroin implies a potential improvement in the barrier function and moisturizing ability of the skin.

Immunofluorescent staining and microscopy:

standard immunofluorescence protocols (based on j.e. gautrot et al, biomaterials33 (2012) 5221-5229) were then used with commercially available primary antibodies against claudin 1, zo-1 and filaggrin (R&D systems, germany) to fix cells and stain. Use of donkey anti-rabbit IgG NorthernLights ^TM NL 637-conjugated antibodies as detected secondary antibodies (R&D systems, germany). Imaging (confocal microscopy) was performed using an SP8 confocal laser scanning microscope (Leica, germany).

Results 3:

as shown in FIG. 3, incubation of HaCAaT cells with 1% OJ showed no up-regulation of tight junction proteins, ZO-1 and filaggrin (untreated vs OJ).

Incubation mixture 1 showed up-regulation of the tight junction protein (untreated vs mixture 1), while the remaining two were not. The oj+nmf combination showed a synergistic upregulation of filaggrin relative to all other test conditions.

Example 4:quantitative measurement of silk fibroin

HACAT cells were cultured in DMEM (Thermo Fisher, art.11995123, germany) supplemented with 10% FCS (Biochrom, merck, germany) at 37℃with 5% CO ₂ Culturing under reduced pressure for 48 hr at inoculation density of 0.5X10 ⁵ Cells/cm ² . Compared to the standard, a sandwich ELISA was performed to quantify the amount of silk fibroin produced in the cells. The concentration of OJ was 1% v/v, and mixture 1 was used at 0.5% w/v (p<0.01). Treatment with 0.5% (w/v) mixture 1 had no effect on the ratio of filaggrin to total protein concentration (0.07). Treatment with OJ did increase this ratio to 0.086. The combination of mixture 1 and OJ (oj+nmf, 1% v/v+0.5% w/v) did increase the filaggrin ratio to 0.096, thus increasing the aggregate synergy by 14%.

Fig. 4 summarizes these results of quantitative measurement of silk fibroin.

Formulation examples

Example 1: O/W emulsion

The procedure is as follows:

xanthan gum is dispersed in the aqueous phase. Phase a and phase B were heated to 75 ℃. Phase a was stirred into phase B. Homogenizing. Cooled to room temperature and phases C and D, buffer and dispersed mixture 1 were added. Stirring until all dissolved.

Suppliers (suppliers)

(1)Seppic；(2)BASF AG；(3)Evonik Nutrition&Care GmbH；(4)Biesterfeld；(5)Merck KGaA,Darmstadt,Germany/EMD Performance Materials；(6)Azelis Germany GmbH；(7)Schülke&May GmbH；(8)Silab

Example 2: face cream (O/W)

The procedure is as follows:

phase a and phase B were combined separately and heated to 80 ℃. Phase a was added to phase B while stirring. Homogenizing. Adding phase C at-60 deg.C. The dissolved phase D was added (warmed to 40-50 ℃). Phase E was added at 40℃and the pH was adjusted to 5.0-5.5 with sodium hydroxide.

The suppliers:

(1)Seppic；(2)IOI Oleo GmbH；(3)BASF SE；(4)Gattefossé(Deutschland)GmbH；(5)Merck KGaA,Darmstadt,Germany/EMD Performance Materials；(6)Silab

example 3: sun protection emulsion with SPF 15

The procedure is as follows:

phase a and phase B are combined separately. Phase B1 was added to B while stirring.

Stirring until a homogeneous phase is obtained. Phase B2 was added and phases a and B were heated to 70-75 ℃.

Phase a was added to phase B without stirring and homogenized.

Cool with stirring, add phase C and adjust pH to 5.5-6.

Homogenizing.

The suppliers:

Merck KGaA,Darmstadt,Germany/EMD Performance Materials；(2)Evonik Nutrition&Care GmbH；(3)BASF SE；(4)R.T.Vanderbilt Company Inc；(5)RAHN GmbH；(6)Symrise；(7)Silab

Claims

1. an active mixture consisting of:

a) A herbal extract obtained from the root of ophiopogon japonicus (Ophiopogon japonicus), said herbal extract being an aqueous solution consisting of 85.90% to 90.10% water, 9.00% to 13.00% ophiopogon root extract, 0.45% to 0.55% 1, 2-hexanediol and 0.45% to 0.55% octanediol, wherein said ophiopogon root extract has 95 to 99% carbohydrates including polysaccharides and 1 to 5% polyphenols;

b) Amino acid: serine, arginine, and histidine;

c) Amino acid derivative: pyrrolidone-5-carboxylic acid; and

d) Calcium lactate hydrate;

wherein serine is present in an amount of 25 to 35% by weight, arginine is present in an amount of 5 to 10% by weight, histidine is present in an amount of 5 to 10% by weight, pyrrolidone-5-carboxylic acid is present in an amount of 10 to 30% by weight, and calcium lactate hydrate is present in an amount of 25 to 35% by weight, wherein the combined total of components b), c) and d) is 100% by weight, and wherein the sum of components a) and b), c) and d) is calculated as 5:1 to 1: 5.

2. The mixture according to claim 1, wherein L-serine, L-arginine, L-histidine, L-pyrrolidone-5-carboxylic acid and L-calcium lactate pentahydrate are used.

3. The mixture according to claim 2, comprising 29 to 31% by weight of L-serine, 7 to 8% by weight of L-arginine, 7 to 8% by weight of L-histidine, 20 to 25% by weight of L-pyrrolidone-5-carboxylic acid and 29 to 31% by weight of L-calcium lactate pentahydrate, wherein the combined total of components b), c) and d) is 100% by weight.

4. The mixture according to claim 2, wherein components b), c) and d) consist of 30.6% by weight of L-serine, 7.5% by weight of L-arginine, 7.3% by weight of L-histidine, 24.2% by weight of L-pyrrolidone-5-carboxylic acid and 30.4% by weight of L-calcium lactate pentahydrate.

5. The mixture according to claim 1, wherein the sum of component (a) and components (b), (c) and (d) is calculated as 3:1 to 1:3 are present in a weight ratio.

6. The reactive mixture of claim 1, wherein the sum of component (a) and components (b), (c) and (d) is calculated as 1:1,2:1,3:1 or 1:3 are present in a weight ratio.

7. Cosmetic and/or dermatological composition comprising:

a) An herbal extract obtained from ophiopogon root, said herbal extract being an aqueous solution consisting of 85.90% to 90.10% water, 9.00% to 13.00% ophiopogon root extract, 0.45% to 0.55% 1, 2-hexanediol and 0.45% to 0.55% octanediol, wherein said ophiopogon root extract has 95 to 99% carbohydrates including polysaccharides and 1 to 5% polyphenols;

b) Amino acid: serine, arginine, and histidine;

c) Amino acid derivative: pyrrolidone-5-carboxylic acid; and

d) Calcium lactate hydrate;

8. Cosmetic and/or dermatological composition according to claim 7, wherein L-serine, L-arginine, L-histidine, L-pyrrolidone-5-carboxylic acid and L-calcium lactate pentahydrate are used.

9. Cosmetic and/or dermatological composition according to claim 8, comprising 29% to 31% by weight of L-serine, 7% to 8% by weight of L-arginine, 7% to 8% by weight of L-histidine, 20% to 25% by weight of L-pyrrolidone-5-carboxylic acid and 29% to 31% by weight of L-calcium lactate pentahydrate, wherein the combined total of components b), c) and d) is 100% by weight.

10. Cosmetic and/or dermatological composition according to claim 8, wherein b), c) and d) consist of 30.6% by weight of L-serine, 7.5% by weight of L-arginine, 7.3% by weight of L-histidine, 24.2% by weight of L-pyrrolidone-5-carboxylic acid and 30.4% by weight of L-calcium lactate pentahydrate.

11. A non-therapeutic method for improving skin dryness associated with aging by topically applying the mixture of claim 1 or the cosmetic or dermatological composition of claim 7 to human skin.

12. A non-therapeutic method for improving the storage of water in keratinocytes and improving the fluidity of water regulated by aquaporin in different layers of keratinocytes by topical application of the mixture according to claim 1 or the cosmetic or dermatological composition according to claim 7 to human skin.