阅读说明：本技术 麦冬提取物、氨基酸、氨基酸衍生物和乳酸钙水合物的协同作用 (Synergistic effect of ophiopogon root extract, amino acid derivative and calcium lactate hydrate ) 是由 安德鲁·菲利普·萨拉扎 乔尔格·冯·哈根 鲁楠 亚历山德拉里·兰 于 2019-09-27 设计创作，主要内容包括：本发明涉及由组分a)、组分b)、组分c)和组分d)组成的活性混合物,所述组分a)是从麦冬(Ophiopogon japonicus)获得的草药提取物,所述组分b)是丝氨酸、精氨酸和组氨酸的混合物,所述组分c)是吡咯烷酮-5-羧酸,并且所述组分d)是乳酸钙水合物,以及其在化妆品和/或皮肤病学组合物中的用途。本发明还涉及化妆品和/或皮肤病学组合物,其包含：a)从麦冬(Ophiopogon japonicus)获得的草药提取物；b)氨基酸：丝氨酸、精氨酸和组氨酸；c)氨基酸衍生物：吡咯烷酮-5-羧酸；和d)乳酸钙水合物。本发明还涉及通过局部施用所述活性混合物或所述化妆品或皮肤病学组合物的非治疗方法。(The present invention relates to an active mixture consisting of a component a) an herbal extract obtained from Ophiopogon japonicus (Ophiopogon japonica), a component b) a mixture of serine, arginine and histidine, a component c) pyrrolidone-5-carboxylic acid and a component d) calcium lactate hydrate, and its use in cosmetic and/or dermatological compositions. The present invention also relates to a cosmetic and/or dermatological composition comprising: a) herbal extracts obtained from radix Ophiopogonis (Ophiopogon japonicus); b) amino acids: serine, arginine, and histidine; c) amino acid derivatives: 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 said cosmetic or dermatological composition.)

1. an active mixture consisting of:

a) herbal extracts obtained from radix Ophiopogonis (Ophiopogon japonicus);

b) amino acids: serine, arginine, and histidine;

c) amino acid derivatives: pyrrolidone-5-carboxylic acid; and

d) calcium lactate hydrate.

2. The mixture according to claim 1, wherein component a) is an herbal extract obtained from the root of Opiophogon japonicus.

3. The mixture according to claim 1 or 2, wherein component a) comprises an enzymatic hydrolysate of the roots of ophiopogon japonicus.

4. Mixture according to one or more of claims 1 to 3, wherein component a) is an extract of the root of Opiophogon japonicus comprising water, 1, 2-hexanediol and octanediol.

5. The mixture according to one or more of claims 1 to 4, wherein component (a) and the sum of components (b), (c) and (d) are present in a ratio of 5: 1 to 1: 5 is present in a weight ratio.

6. Use of the active mixture according to claim 1 for the preparation of a cosmetic and/or dermatological composition.

7. Cosmetic and/or dermatological composition comprising:

a) herbal extracts obtained from ophiopogon japonicus;

b) amino acids: serine, arginine, and histidine;

c) amino acid derivatives: pyrrolidone-5-carboxylic acid; and

d) calcium lactate hydrate.

8. The cosmetic and/or dermatological composition according to claim 7, wherein component a) is an herbal extract obtained from the roots of Ophiopogon japonicus.

9. The cosmetic and/or dermatological composition according to claim 7 or 8, wherein component a) comprises an enzymatic hydrolysate of the root of Opiophogon japonicus.

10. The cosmetic and/or dermatological composition according to one or more of claims 7 to 9, wherein component a) is an extract of the roots of ophiopogon japonicus comprising water, 1, 2-hexanediol and caprylyl glycol.

11. Cosmetic and/or dermatological composition according to one or more of claims 7 to 10, wherein the sum of component (a) and components (b), (c) and (d) is such that the ratio of 5: 1 to 1: 5 is present in a weight ratio.

12. A non-therapeutic method of ameliorating age-related skin dryness by topically applying to human skin the mixture of claim 1 or the cosmetic or dermatological composition of claim 7.

13. A non-therapeutic method for reducing or preventing chronic inflammation and irritation associated with aging by topically applying to human skin a mixture according to claim 1 or a cosmetic or dermatological composition according to claim 7.

14. A non-therapeutic method for stimulating the synthesis of hyaluronic acid, aquaporins and filaggrin in living cells of the epidermis and dermis by topically applying to human skin a mixture according to claim 1 or a cosmetic or dermatological composition according to claim 7.

Technical Field

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

a) herbal extracts obtained from radix Ophiopogonis (Ophiopogon japonicus);

b) amino acids: serine, arginine, and histidine;

c) amino acid derivatives: 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 said cosmetic or dermatological composition.

Background

The epidermis is the outermost layer of the skin and is composed primarily of keratinocytes (keratinocytes). These cells differentiate upward to form a basement membrane (basal layer) where they contact the dermis, forming stratified layers (e.g., spinous and granular layers) that ultimately serve as a sheet of connected keratinocytes at the outermost position, the Stratum Corneum (SC). Each stage of epidermal differentiation is characterized by the expression of a specific protein.

Article number 15900(2017), page 1-13, by Chunsik Choe et al, Nature Scientific Reports, Vol.7, describes that SCs are considered skin barriers that prevent foreign substances from penetrating the skin and also prevent moisture from evaporating 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 α -helical conformational form. Thus, water cannot be inserted into the keratin chain and there are no vacancies for water molecules to bind to the keratin. It was further found that the natural moisturizing factor NMF is primarily responsible for binding water in these SC layers. In the intermediate layer (70-30% SC depth), the keratin chains are more spread and, therefore, the interactions between the keratin side chains are weak and water molecules are embedded in the keratin filaments and bind there, causing the keratinocytes (corneocytes) to swell. At the bottom layer (100-80% SC depth), the keratin chain has fewer vacancy binding sites than the middle layer because the keratin folds are greater than at 70% SC depth, with the lowest NMF concentration and the greatest amount of water. Thus, in this SC layer, the incoming water cannot effectively form hydrogen bonds with the surrounding keratin and NMF, which makes this SC layer the bottommost low swelling 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 significant changes in the composition and functional capacity of the epidermis.

To prevent diseases associated with dry skin, impaired barrier function or redness, the traditional approach has been to focus on "hydration" of the SC. However, the silk fibroin-related biochemical processes also appear to play an important role.

Aileen Sandilans et al, "Filaggrin in the frontline: roll in skin barrier function and disease ", Journal of Cell Science122(2009), 1285-1294, describes that the deletion of profilaggrin or filaggrin results in the formation of a poor stratum corneum, which is prone to water loss. It is further pointed out that recent human genetic studies strongly suggest that perturbation of skin barrier function leads to enhanced transdermal transfer of allergens due to reduced or complete loss of filaggrin expression.

Natural Moisturizing Factor (NMF) is a product of degradation of filaggrin in SC and SG. The modified filaggrin results in a reduction of NMF in the Stratum Corneum (SC) and Stratum Granulosum (SG) layers.

Nobutomo Ikarashi et al, "Relationship between Aging and related skin dryness and aquaporins", International Journal of Molecular Sciences, 2017,18,1559, describe a study that analyzes the mechanisms of Aging-related skin dryness, focusing on aquaporins (aquaporins), which are water channels. The research result shows that the expression level of the aquaporin in the skin is reduced along with the aging, which indicates that the aquaporin can be a new target point for preventing and treating the aging-related dry skin.

Thus, a method of skin dryness and improvement of barrier function can be addressed by stimulating skin cells to synthesize macromolecules involved in these processes, such as filaggrin or aquaporin.

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.

Ophiopogon japonicus (also known as Dwarf lilyturf, Farmula, Foundatioplant, Monkeygrass, Snake whiskers) is a species of Ophiopogon, native to China, India, Japan and Vietnam. It is an evergreen, perennial plant that forms turf. The leaves are linear and 20-40 cm long. The flower is a pale white, light purple. Underground, this species has large stolons and tuberous roots.

In traditional Chinese medicine, the tuber root of dwarf lilyturf is called dwarf lilyturf tuber and is the main herb for yin deficiency. According to Chinese Herbal Medicine Materia Medica, the herb is sweet in taste, slightly bitter and cold, enters heart, lung and stomach channels, nourishes yin and stomach, and has the functions of nourishing spleen, heart and lung, clearing heat, calming, removing irritability, moistening intestinal tract, strengthening stomach function and nourishing lung. Some of the main components of this herb are reportedly stigmasterol, mucus, saponins, carotenoids, steroid saponins and polysaccharides.

In Pengjiao Zeng et al, Progress in Molecular Biology and Molecular Science, 2019,3.6 "Ophiogon japonica" describes that polysaccharides are the main components of Ophiopogon japonicus, with an extraction rate of 35%. Molecular weights of ophiopogonpolysaccharides are not consistently described, ranging from 2.74 to 325kDa, consisting mainly of β -fructose and small amounts of α -glucose. Fructans are polymers of fructose molecules.

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

US20110045105 describes the use of an extract of ophiopogon japonicus as active ingredient, which preferably acts by increasing the NMF level of the stratum corneum, in particular by increasing the levels of lactate and serine. It further describes the cosmetic use of an extract of Ophiopogon japonicus 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 claudin-1 and zonula occludens ZO-1.

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

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

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

US2008/0260672 describes an aqueous preservative solution having a high amino acid content, which has a particular preservative effect against fungi, which has excellent storage stability, and when incorporated into cosmetics, is capable of imparting a high moisturizing effect without causing a sticky feeling, and has an effect of preventing the discoloration of dyed hair. 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 at a specific pH.

WO2008/142147 describes a composition comprising pyrrolidone-5-carboxylic acid and at least one compound from citrulline, arginine and asparagine in racemic or isomeric form 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 its salts, pyrrolidone carboxylic acid or its salts and amino acids for controlling the pH of the skin, and cosmetic compositions comprising the same. Such compositions are also reported to protect skin barrier function, and the ingredients of the compositions are part of Natural Moisturizing Factor (NMF). As a result, such compositions or cosmetics comprising such compositions are reported to be capable of moisturizing the skin.

EP2404502 describes compositions containing pyrrolidone carboxylic acid and a metal salt such as a copper, zinc or manganese salt, 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 is still a need to find a powerful solution to improve the skin dryness associated with aging and to reduce or prevent the 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 present inventors have now found that the above objects can be achieved by the process described herein or by the specific mixtures of the present application.

The invention relates to an active mixture consisting of:

a) herbal extracts obtained from radix Ophiopogonis (Ophiopogon japonicus);

b) amino acids: serine, arginine, and histidine;

c) amino acid derivatives: pyrrolidone-5-carboxylic acid; and

d) calcium lactate hydrate.

The invention also relates to the use of the active mixture for preparing cosmetic and/or dermatological compositions.

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

a) herbal extracts obtained from radix Ophiopogonis (Ophiopogon japonicus);

b) amino acids: serine, arginine, and histidine;

c) amino acid derivatives: pyrrolidone-5-carboxylic acid; and

d) calcium lactate hydrate.

The invention also relates to a non-therapeutic method for ameliorating the dryness of the skin associated with ageing or for reducing or preventing the chronic inflammation and irritation associated with ageing by topically applying the 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 a cosmetic composition comprising said components of said mixture.

Drawings

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

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

Figure 3 shows immunofluorescence staining of epidermal barrier protein as described in example 3.

Figure 4 shows quantitative measurements of filaggrin as described in example 4.

Detailed Description

"active mixtures" in the sense of the present invention means mixtures 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 feed is composed of

a) Herbal extracts obtained from radix Ophiopogonis (Ophiopogon japonicus);

b) amino acids: serine, arginine, and histidine;

c) amino acid derivatives: pyrrolidone-5-carboxylic acid; and

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

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

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

-dissolving the tuber of dwarf lilyturf powder in water,

-at least one enzymatic hydrolysis,

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 its reaction parameters based on his general knowledge.

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 carbohydrase. 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 fructan, relative to the total weight of dry matter, preferably at least 80%. Preferably, the fructose content of the material is from 28 to 245g/l, preferably from 85 to 127 g/l. Fructose was metered spectrophotometrically according to the method for determining ketose described in Boratynski, J., Analytical Biochemistry, 1984,137,2, 528-532.

The preferred aqueous extract of ophiopogon root has 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 the root of Opiophogon japonicus, which comprises water, 1, 2-hexanediol and caprylyl glycol.

Component a) is particularly preferably an aqueous solution consisting of 85.90% to 90.10% water, 9.00% to 13.00% of the extract of Ophiopogon japonicus root as described above, 0.45% to 0.55% 1, 2-hexanediol and 0.45% to 0.55% caprylyl glycol, under the trade name "Hemophilus" (R.TM.)Marketed by Silab company, France.

The invention also relates to the use of said active mixtures as described above or of active mixtures having components a), b), c) and d) as described above or below, preferably in the preparation of cosmetic and/or dermatological compositions.

The cosmetic and/or dermatological composition may comprise the 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 previously or preferably before or after it.

In the cosmetic and/or dermatological composition, the total weight of components a), b), c) and d) is comprised between 0.1 and 20% by weight, preferably between 0.5 and 10% by weight, more preferably between 1 and 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. Preferred calcium L-lactate hydrate is calcium L-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 to use 0.3 to 0.4 equivalents of calcium lactate hydrate together with 1 equivalent of serine. When used according to the invention, it is particularly preferred to use 1: 0.33 molecular ratio 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 cosmetic or dermatological composition according to the invention.

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

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

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

The following amounts of the components are particularly preferred: l-serine 29 to 31% by weight, L-arginine 7 to 8% by weight, L-histidine 7 to 8% by weight, 20 to 25% by weight, L-pyrrolidone-5-carboxylic acid and 29 to 31% by weight of L-calcium lactate pentahydrate, wherein the combined total amount 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 previously or preferably as described previously to construct a preferred active mixture according to the invention.

The active mixtures as described or preferably as described above preferably contain components a) and the sum of 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.

the cosmetic and/or dermatological compositions as described or preferably as described above preferably contain a sum of components a) and 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 generally cosmetic or dermatological formulations, which may be applied topically. By "topically applicable" 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, depending on the desired property profile, optionally also other suitable ingredients. The topical formulations are preferably used as cosmetic compositions. Suitable carriers and adjuvants or fillers are known in the art.

The preparations 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-preventing agents, irritation-inhibiting agents, antioxidants, astringents, perspiration-inhibiting agents, preservatives, antistatic agents, binders, buffers, carrier materials, chelating agents, cell-stimulating agents, cleaning agents, care agents, depilatory agents, surfactants, deodorants, antiperspirants, emollients, emulsifiers, enzymes, essential oils, fibers, film-forming agents, fixatives, foam-forming agents, foam stabilizers, foam-preventing substances, foam boosters, gelling agents, gel-forming agents, hair care agents, hair-setting agents, hair-straightening agents, moisturizers, moisturizing substances, bleaching agents, reinforcing agents, dye-removing agents, optical brighteners, impregnating agents, anti-soiling agents, friction-reducing agents, lubricants, opacifiers, plasticizers, coating agents, polishing agents, shine agents, polymers, proteins, re-lubricants, abrasives, silicones, skin soothing agents, skin cleansers, skin protectants, skin rejuvenating agents, skin lightening agents, skin care agents, skin softeners, hair boosters, cooling agents, skin cooling agents, warming agents, skin warming agents, stabilizers, uv absorbers, organic uv filters, inorganic uv 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 protection agents, pigments, preservatives, fragrances, flavoring substances, odorous substances, polyols, surfactants or electrolytes.

Particularly suitable active ingredients for the combination are ectoin (A), (B), (C), (Ectoin), hydroxyectoin, trehalose, glycerol, glycosylglycerol, β -mannosylglycerol (firoin), β -mannosylglycamide (firoin A), di-phosphoinositide (DIP), cyclic 2, 3-diphosphoglyceric acid (cDPG), 1, 1-diglycerol phosphate (DGP), di-mannosylinositol Diphosphate (DMIP), betaine, glycine betaine, proline betaine, glutamic betaine, alanine, proline, glutamine, N-acetyl lysine, glutamine 1-amide, taurine, choline O-sulfate, carnitine, arsenic betaine, crotonobetaine, dimethyl sulfoacetate, dimethyl sulfonate, high betaine, trimethylamine N-oxide, panthenol, sorbitol, meglumine, hyaluronic acid or hyaluronic acid derivatives, ureaUrea) and nicotinamide (Niacinamide), 5, 7-dihydroxy-2-methylchromone, tradenameMarketed or commercial productsRenouMer，VTA，Poppy SE，Isoquercetin orCyclopeptide 5，The Cyclopeptide 5 is alcohol-free,SereneShield，Emblica，orA liquid.

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

These further active ingredients are preferably present in the topical formulations 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 in the UVA region and/or the UVB region and/or the IR and/or VIS region (absorbers). These substances can be chosen in particular from dibenzoylmethane derivatives, cinnamic acid derivatives, salicylic acid derivatives, camphor derivatives, triazine derivatives, beta-diphenylacrylate derivatives, p-aminobenzoic acid derivatives and polymeric and silicone filters. The UV filters are generally named according to INCI nomenclature as follows.

Particularly suitable for combination with the active mixtures according to 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, terephthaloyldicamphor sulfonic acid, disodium phenyldibenzoimidazole tetrasulfonic acid, methylenebis (benzotriazolyl) tetramethylbutylphenol, butylmethoxydibenzoylmethane, ethylhexyltriazone, diethylhexylbutamidotriazone, triazolocyclotrisiloxane, phenylenebiphenyltriazine, polysiloxane-15, 1, 1-dicarboxyl (2,2' -dimethylpropyl) -4, 4-diphenylbutadiene, 2, 4-bis [5-1 (dimethylpropyl) benzoxazol-2-yl (4-phenylbistriazine) ) Imino ] -6- (2-ethylhexyl) imino-1, 3, 5-triazine, and mixtures thereof. These organic UV filters are generally incorporated in the formulations in amounts 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 alkylbenzoate, isopropyl myristate, arylalkyl benzoate, e.g. phenethylbenzoate (X-Tend 226) or cosmecol brand oil components such as dimyristol tartrate, tri C14-C15 alkyl citrate, C12-C13 alkyl lactate, tridecyl salicylate, C12-C13 alkyl octanoate, 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 solubilizers (e.g. butylphthalimide, isopropylphthalimide, dimethyl isosorbide.) very particularly preferred cosmetic oils are C12-C13 alkyl lactate (available under the trade name cosmecol ELI), and phenethyl phenylacetate (available under the trade name X-Tend 226).

The 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 can be applied or sprayed onto the skin, for example as creams or milks (O/W, W/O, O/W/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, face cleansers or essences.

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

Preferred adjuvants are from the group of preservatives, stabilizers, solubilizers, colorants, odor improvers, thickeners, plasticizers, humectants, surfactants, emulsifiers, preservatives, antifoams, 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 conventional carriers such as lactose, talc, silica, aluminum hydroxide, calcium silicate and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary volatile liquefied propellants, for example chlorofluorohydrocarbons, propane/butane or dimethyl ether. Compressed air may also be advantageously used.

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

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

The facial and body oils may contain 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, paraffin oils, lanolin oils or mixtures of these substances.

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

Preferred formulation forms also include in particular 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 coemulsifiers in the preferred O/W emulsions.

Preferred oils and/or lipids for use in the topical formulation include: paraffin wax, 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 kernel oil (Prunus Cerasus), avocado oil, safflower seed oil, macadamia nut oil, cocoa butter (Theobroma Cacao), shea butter and mixtures thereof.

Preferred absorbing and/or thickening agents for use in the formulations include modified corn starch, silica, talc, zinc stearate, magnesium sulfate, zinc oxide, calcium and aluminum borosilicates, starch and derivatives, polyurethanes, and mixtures thereof.

The active mixture as described previously or preferably as described previously synergistically stimulates the synthesis of hyaluronic acid by HAS1, synergistically stimulates the synthesis of the aquatransporter 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 the dryness associated with skin aging and synergistically enhance the barrier function of the epidermis.

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

In another 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 topically applying to the human skin an active mixture as described previously or by topically applying a cosmetic or dermatological composition comprising an active mixture as described previously or a cosmetic or dermatological composition comprising components a), b), c) and d) as described previously. Preferably, the non-therapeutic method is a cosmetic method of applying a cosmetic composition.

This application is carried out using standard techniques, for example by applying shampoos, shower gels, creams, face washes, pastes, gels, lotions, essences to the skin to be treated, or dissolving a predetermined amount of a preparation comprising an active mixture as described previously or an active mixture as described previously 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 specification may be replaced by an alternative feature serving the same purpose, or an equivalent or similar purpose, unless expressly excluded. Thus, unless expressly stated otherwise, any feature disclosed in this specification is to be considered as an example of a generic series of equivalent or similar features.

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

The technical teachings disclosed herein may be abstracted away and combined with other examples.

The complete disclosures of all applications and publications mentioned above and below are incorporated by reference into this application.

The following examples are intended to illustrate the invention. However, they should in no way be considered limiting.

Materials:

the components b), c) and d) used in this example consisted of the following ingredients disclosed in table 1. Said 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 was used at a concentration of 1%.

Table 2:

the active mixture used in the examples was 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.111995123, Germany) supplemented with 10% FCS (Biochrom, Merck, Germany) were seeded onto coverslips in 6-well plates at 37 ℃ with 5% CO₂In the middle by 0.5 × 10⁵Individual cell/cm²Cultured at the density of (1) for 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 compounds could increase proteins that play a role in the synthesis of macromolecules involved in skin moisture retention. The key molecule in skin retention is hyaluronic acid. This molecule is synthesized by hyaluronic acid synthase 1(HAS-1) and hyaluronic acid synthase 2 (HAS-2).

Gene expression analysis was performed using primers from a universal probe library in the form of a custom assay provided by the realme Ready system (Roche, Manheim, germany). For this purpose, with the aid of the RNeasy Mini Kit (QIAGEN, Hilden, Germany)RNA was isolated. cDNA was synthesized from the isolated RNA using a High-Capacity cDNA Reverse Transcription kit with RNase inhibitor kit (Thermo, Darmstadt, Germany). Finally, gene expression was analyzed using LC480(Roche, Manheim, germany) according to the manufacturer's recommended cycling conditions. Use 2^-ΔΔCtThe method calculates fold change relative to control.

Results 1:

as shown in fig. 1, untreated HaCaT cells did produce basal levels of HAS-1, which was set at 1 for reference. Treatment with OJ up-regulated HAS-1RNA by 58%, while the active cocktail of OJ and cocktail 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, Art 11995123, Germany) supplemented with 10% FCS (Biochrom, Merck, Germany) were seeded onto coverslips in 6-well plates at 37 ℃ with 5% CO₂In the middle by 0.5 × 10⁵Individual cell/cm²Cultured at the density of (1) for 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 effects on the storage of water in the cells and the mobility of water in different layers of the cells (regulated by aquaporins) and quantified the expression profile of the aquaporin family.

As shown in figure 2, up-regulation of aquaporin 1 by 161% was detected by treatment with OJ, and the active mixture of OJ + NMF increased expression to 233%. UT is an 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.111995123, Germany) supplemented with 10% FCS (Biochrom, Merck, Germany) were seeded onto coverslips in 6-well plates at 37 ℃ with 5% CO₂In the middle by 0.5 × 10⁵Individual cell/cm²Cultured at the density of (1) for 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 the skin barrier proteins claudin, zonula occludens and filaggrin 48 hours after treatment of the active mixture OJ + NMF. The increase in claudin and zonules closed indicates improved binding of keratinocytes to each other. Whereas, since degradation of filaggrin produces natural moisturizing factors, the increase of filaggrin means a potential improvement in the barrier function and moisturizing ability of the skin.

Immunofluorescence staining and microscopy:

standard immunofluorescence protocols (based on J.E. Gautrot et al, Biomaterials33(2012)5221-&D systems, germany) to fix the cells and stain. Use of donkey anti-rabbit IgG northern lights^TMNL 637-conjugated antibodies as Secondary antibodies (R) for detection&D systems, germany). Imaging (confocal microscopy) was performed using SP8 confocal laser scanning microscopy (Leica, germany).

Results 3:

as shown in FIG. 3, incubation of HaCAaT cells with 1% OJ showed no upregulation of claudin, ZO-1 and filaggrin (untreated vs OJ).

Incubation mixture 1 showed upregulation of claudin (untreated vs mixture 1), the other two did not. The combination of OJ + NMF showed a synergistic up-regulation of filaggrin relative to all other tested conditions.

Example 4:quantitative measurement of filaggrin

HACAT cells were cultured in DMEM (Thermo Fisher, Art 11995123, Germany) supplemented with 10% FCS (Biochrom, Merck, Germany) at 37 ℃ with 5% CO₂Culturing for 48 hr at a seeding density of 0.5 × 10⁵Cells/cm². Sandwich ELISA for quantification of production in cells compared to standardsAmount of raw silk fibroin. OJ concentration 1% v/v, mixture 1 was used at 0.5% w/v (p)<0.01). Treatment with 0.5% (w/v) of mixture 1 had no effect on the ratio of filaggrin to total protein concentration (0.07). Treatment with OJ did increase the 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 adding up to a 14% synergistic increase.

Fig. 4 summarizes these results for quantitative measurements of filaggrin.

Formulation examples

Example 1: O/W emulsion

The procedure is as follows:

dispersing xanthan gum in the aqueous phase. Phase A and phase B were heated to 75 ℃. Stir phase a into phase B. And (6) homogenizing. Cool to room temperature and add phases C and D, buffer and dispersed mixture 1. Stirring until all dissolved.

Suppliers of goods

(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:

combine phase A and phase B separately and heat to 80 ℃. Phase A was added to phase B while stirring. And (4) homogenizing. Phase C was added at-60 ℃. Add dissolved phase D (warm to 40-50 ℃). Phase E was added at 40 ℃ and the pH was adjusted to 5.0-5.5 with sodium hydroxide.

The supplier:

(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: sunscreen emulsion with SPF 15

The procedure is as follows:

combine phase A and phase B separately. Phase B1 was added to B while stirring.

Stirring until a homogeneous phase is obtained. Add phase B2 and heat phase a and B to 70-75 ℃.

Add phase a to phase B without stirring and homogenize.

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

And (4) homogenizing.

The supplier:

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。