EP2032119A1

EP2032119A1 - Cosmetic preparation with aquaporin stimulators and the use thereof

Info

Publication number: EP2032119A1
Application number: EP07727450A
Authority: EP
Inventors: Wilfried Siefken; Ute Breitenbach; Tanja Delekat; Lara Terstegen; Helga Biergiesser; Franz STÄB; Gunja Springmann
Original assignee: Beiersdorf AG
Current assignee: BASF Beauty Care Solutions France SAS
Priority date: 2006-04-27
Filing date: 2007-03-28
Publication date: 2009-03-11
Also published as: CN101453979B; US20090130223A1; CN101453979A; WO2007124991A1

Abstract

The development relates to cosmetic and dermatological preparations comprising aquaporin stimulators, especially glyceryl glycosides, and the use thereof for improving moisturization of the skin.

Description

Beiersdorf AG

Cosmetic preparation with aquaporin stimulators and their use

The development relates to cosmetic and dermatological preparations containing aquaporin stimulators, in particular glyceryl glycosides, and their use for improving skin moisturization.

The skin is the largest organ in humans. Among its many functions (for example, for heat regulation and as a sense organ) is the barrier function, the one that prevents the drying of the skin (and thus ultimately the entire organism), probably the most important. At the same time, the skin acts as a protective device against the penetration and absorption of substances coming from outside. This barrier function is caused by the epidermis, which, as the outermost layer, forms the actual protective cover against the environment. At about one-tenth of the total thickness, it is also the thinnest layer of the skin.

The epidermis is a stratified tissue in which the outer layer, the stratum corneum, represents the part of the barrier function. It is worn out in contact with the environment and is therefore in a constant process of renewal, whereby outwardly continuously fine scales are released and from the inside keratinized cell and lipid material is reproduced.

Elias Skin Model (PM Elias, Structure and Function of the Stratum Corneum Permeability Barrier, Drug Dev. Res. 13, 1988, 97-105), which is now recognized in the art, describes the horny layer as a two-component system, similar to a brick wall (US Pat. Brick mortar model). In this model, the horny cells (corneocytes) correspond to the bricks, the complex lipid membrane in the intercellular spaces corresponds to the mortar. This system is essentially a physical barrier against hydrophilic substances, but due to its narrow and multi-layered structure, it is also difficult to pass by lipophilic substances. The special structure of the horny layer protects the skin and stabilizes its own flexibility by binding a defined amount of water.

Even mechanical loads, such as pressure, shock or shear forces, can to a surprising extent by the horny layer alone or in conjunction with the deeper Hautschich- be intercepted. Greater pressure, turning or shearing forces are passed on to the deeper skin layers via the interlocking of the epidermis with the dermis (papilla structure).

The regulation of water and moisture content is one of the most important functions of the epidermal lipid membrane. However, it not only has a barrier effect against external chemical and physical influences, but also contributes to the cohesion of the horny layer.

The lipids of the horny layer consist essentially of ceramides, free fatty acids, cholesterol and cholesterol sulfate and are distributed over the entire horny layer. The composition of these lipids is crucial for the intact functioning of the epidermal barrier and thus for the impermeability of the skin.

Even when the skin is cleansed with the aid of a simple water bath - without the addition of surfactants - the horny layer of the skin initially swells. The degree of this swelling depends, among other things, on the duration of the bath and its temperature. At the same time water-soluble substances are washed off or washed out, such as. As water-soluble soil constituents, but also the skin's own substances that are responsible for the water-binding capacity of the horny layer. In addition, dermal fat is also loosened and washed out to a certain extent by the skin's own surfactants. This causes after initial swelling a subsequent dehydration of the skin, which can be significantly enhanced by detergent additives.

In healthy skin, these processes are generally irrelevant, as the protective mechanisms of the skin can readily compensate for such minor disorders of the upper layers of the skin. But even in the case of non-pathological deviations from the normal status, eg. B. by environmental wear damage or irritation, photodamage, aging skin, etc., the protective mechanism is disturbed on the skin surface.

In the case of old skin, for example, the regenerative renewal slows down, whereby in particular the water-binding capacity of the horny layer subsides. It therefore becomes inflexible, dry and cracked ("physiologically" dry skin). A barrier damage is the result. The skin becomes susceptible to negative environmental influences such as the invasion of microorganisms, toxins and allergens. As a result, even toxic or allergic skin reactions may occur. In the case of pathologically dry and sensitive skin, barrier damage is a priori. Epidermal intercellular lipids are formed incorrectly or in insufficient quantity or composition. The consequence is an increased permeability of the horny layer and insufficient protection of the skin from loss of hygroscopic substances and water.

The barrier effect of the skin can be quantified by determining transepidermal water loss (TEWL). This is the evaporation of water from inside the body without the loss of water during sweating. The determination of the TEWL value has proven to be extraordinarily informative and can be used for the diagnosis of chapped or chapped skin, for the determination of the compatibility of chemically different surfactants and the like.

For the beauty, neatness and natural function of the skin, the water content in the uppermost skin layer is of the utmost importance. It can be favorably influenced to a limited extent by introducing moisture regulators (moisturizers), for example glycerol in cosmetic formulations.

Anionic surfactants, which are generally components of cleansing formulations, can increase the pH in the horny layer for a long time, severely hampering regenerative processes that serve to restore and renew the barrier function of the skin. In this case, in the horny layer between regeneration and the loss of essential substances by regular extraction, a new, often very unfavorable equilibrium state, which significantly affects the appearance of the skin and the physiological function of the horny layer.

Products for the care, treatment and cleaning of dry and stressed skin are known per se. However, their contribution to the regeneration of a physiologically intact, hydrated and smooth horny layer is extensive and limited in time.

The effect of ointments and creams on the barrier function and the hydration of the horny layer is based essentially on the coverage (occlusion) of the treated skin areas. The ointment or cream is, so to speak, a (second) artificial barrier that is supposed to prevent the loss of water from the skin. Accordingly, this physical barrier can be removed again, for example with cleaning agents, whereby the original, impaired state is again achieved. In addition, the skin care effect can be relax moderately. Furthermore, cosmetic formulations are usually added to a moisturizer. Moisturizers are skin-friendly, hygroscopic substances (eg glycerine, urea or amino acids) that are supposed to retain the water that evaporates from the skin. After discontinuation of the product application, the skin returns to its pre-treatment state very quickly. For certain products, the condition of the skin may even temporarily deteriorate. A sustainable product effect is usually not achieved or only to a limited extent.

Water transport across cell membranes is a fundamental process of life that has received strong attention during the last century. Awareness of its physiological and clinical significance has intensified following the discovery of a specific water channel in red blood cells by Peter Agre. Peter Agre was awarded the Nobel Prize for Chemistry in 2003.

Water is central to the function of the skin. In addition to the maintenance of all transport tasks and physiological functions in the living layers of the epidermis (eg, stratum basale, S. spinosum, S. granulosum), water is also of great importance in the stratum corneum. The enzymes which are active there can also only fulfill their functions to a sufficient extent if the degree of hydration of the S. corneum is sufficient. In particular, the correct pH is a prerequisite for enzyme activities.

Applied exogenously, applied externally, glycerin is a cheap moisturizer. Moisturizers (moisturizers) are not moisturizers (humectants) in the true sense, but substances or mixtures that give cosmetic agents the property to increase after slight massaging into the skin, the moisture content of the horny layer (stratum corneum). As moisturizers are recommended: arginine pyroglutamate, chondroitin sulfate, hvaluronic acid, inositol, lactic acid (sodium lactate), sodium acrylate-vinyl alcohol copolymers, sodium isostearyl-2-lactate, oligopeptides, polysiloxanes, pyroglutamic acid, 2-pyrrolidone and uronic acids. Moisturizers have no occlusive effect in contrast to the likewise moisture-increasing Vaseline. The effectiveness of a moisturizer can be determined by determining the TransEpidermal Water Loss (TEWL)

As a moisturizer, glycerin also provides enhanced hydration of the stratum corneum through its water-binding properties. Endogenously, glycerol is not only a moisturizer, but also a metabolite important for triglyceride synthesis. Glycerol also provides an energy source in the metabolism of cells.

Aquaporins represent a group of structurally related proteins occurring in plant and animal cell membranes which form channels (pores) for polar substances of low molecular weight, especially water.

Aquaporins allow the rapid exchange of larger amounts of water and glycerol. through the plasma membrane and intracellular membranes, e.g. In erythrocytes, epithelial cells or growing plant cells. In contrast to uncatalyzed, purely physical diffusion through the lipid layer, in aqueducts the aquaporin-mediated transport of water through the plasma membrane is characterized by lower sensitivity to low temperatures and inhibitivity by inhibitors (eg HgC ^). From a functional point of view, the group of aquaporins includes the TIP proteins (TIP = tonoplast intrinsic protein) and PIP proteins (PIP = plasma membrane intrinsic protein) from plant cells and the CHIP proteins (CHIP = channel-forming integral protein) from the plasma membrane animal cells. The expression of cDNAs of the TIP, PIP or CHIP genes in Xenopus oocytes (amphibious oocytes, Xenopus ovoc expression systems) considerably increases the water exchange through the plasma membrane of these cells - a strong support for the water transport function of these proteins , From a genetic point of view, the TIP, PIP and CHIP proteins in turn belong to an evolutionary family of channel-forming membrane proteins, MIP proteins (MIP = major intrinsic protein) and possess 6 membrane-spanning domains. They are tetrameric in the membrane.

In many organs aquaporins play an important role in the regulation of the water balance. They prevent the cells from bursting (osmotic regulation), for example when the salt concentration in the environment changes. Thus, the primary urine secretion and secondary urine formation in the kidney with the help of aquaporins. The secretion of some exocrine glands (salivary gland, lacrimal gland) also involves a significant amount of aquaporins.

DE 199 44 625 describes antiperspirant preparations containing modulators of aquaporins. However, the task and effect of the aquaporin modulators is not explained. However, since they are antiperspirant preparations, ie preparations which reduce the escape of fluid from the skin pores (sweating) or It should be assumed that the modulation is the control of the water transport of the cells among each other and not the stimulation of aquaporin expression, ie an increase in the number of aquaporins.

Taken together with aquaporins from plants, bacteria, amphibians, etc., there are more than 150 isoforms. The functional classification of aquaporins has so far provided two groups:

a) pure water pores (aquaporins: AQP-O, 1, -2, -4, -5, -6, and -8) and b) pores, which in addition to water also allow small uncharged molecules such as glycerol and urea to pass through (aquaglyceroporins: AQP -3, -7, -9 and -10).

In AQP-3-less mice, it has been shown that the glycerol content of the skin is reduced (Hara, Ma and Verkmann in J Biol. Chem. 277, 46616-46621) and leads to a deficient hydration of the stratum corneum. In addition, in these mice, the skin elasticity is lowered and the barrier repair after stratum corneum damage slows down. The water content in the stratum corneum of the AQP-3-less mice is reduced by a factor of three, which correlates with the decreased glycerol content (also factor three). This is a clear indication that the water-binding capacity of glycerol is essential for stratum corneum humidification.

The skin is able to slowly adapt to dry ambient conditions (eg winter climate, air conditioning systems) through increased ceramide synthesis and thus counteract dehydration. However, modern living conditions (eg artificial indoor climate, extensive body cleansing) can dramatically reduce the functionality of this natural mechanism.

It is known that the use of marine minerals (bathing in the "dead sea") leads to an improvement in the condition of the skin or that the application of marine mineral-containing cosmetic formulations increases the lipid barrier of the skin Cell culture model shows that the osmotic stress caused by increased salinity of the culture medium and thus increased osmolarity, causes an increase in AQP-3 expression.This increase in aquaglycero-porins suggests a protective / re-balancing reaction of the skin as a "countermeasure" This "in vitro" simulated dryness will close and ultimately result in better moisturization of the skin Inside and an improved absorption capacity of offered in a cosmetic or dermatological preparation glycerin and water.

In particular, dry skin suffers from low levels of water and glycerol in the upper epidermis layers, including the stratum corneum. Dry skin is often caused by exogenous factors, such as. As stress conditions (UV radiation, winter climate, dry indoor climate, for example, by air conditioning) or by endogenous factors such. As skin aging and atopy, caused.

Important enzymes, such as. B. necessary for controlled desquamation of the cornea, work to the extent necessary only with sufficient degree of hydration and specific pH of the ambient environment. The result of insufficient enzyme activity in this case could be a flaky, prone to itching, even visually disturbed skin.

The water transport from the deeper skin layers upwards is limited. The water and glycerol transport must be through the cell membranes, this is the responsibility of aquaporins. The number of aquaporins present in the cell membranes of the skin is limited and differs depending on the skin type and skin region.

It is therefore necessary to treat certain skin areas, in particular the horny layer, with moisturizing cosmetic and dermatological preparations. Conventional cosmetics only affect the causative loss of water, by occlusion and supply of lipids to improve the barrier of the cornea, as well as the administration of moisturizers such. As glycerol or urea, contrary. The effect achieved thereby is usually only for a short time, since in general no depth effect, d. H. no moistening deeper skin layers, is achieved.

An increase in aquaporin expression is possible in the prior art only through the use of steroids. Are known steroids, such as. For example, the Ecdvsteroid from Ajupa Turkestanica, which causes the formation of aquaporins in the cell membranes via hormonal stimulation of cell metabolism.

However, steroids are not suitable for cosmetic products due to their multitude of side effects. According to the invention, therefore, aquaporin stimulators are used which are not too the steroids count, so no cyclopentanoperhydrophenanthrene skeleton (gonan scaffold) have.

The prior art therefore lacks preparations which promote or positively influence the body's own improvement in the hydration of the horny layer without having harmful side effects.

Based on this known prior art, it is an object of the invention to positively influence the moisture balance of the skin.

It was unforeseeable for the skilled worker that a promotion and stimulation of aquaporin expression and thus an increase of the body's own or endogenous and exogenous supply of the skin with water and moisturizers, such as glycerol, by cosmetic and dermatological preparations containing glyceryl glycosides, is possible ,

This increased amount of absorbed and better bioavailable "moisture" is released from the cells over time and leads to an improved hydration or physiological function of the upper epidermis layers.These improvements are characterized, among others, by:

improved homeostasis (enzyme activities, nutrient delivery-waste removal), improved elasticity (wrinkle reduction), improved protection against infections, improved skin feel (reduced tension, cracking, itching) and - improved energy supply

It was also an object of the present invention to provide skin-care preparations which retain or restore the barrier properties of the skin, especially when the natural hydration, in particular of the dry skin, is insufficient.

They should also be suitable for the prophylaxis of consequential damage of skin dehydration, such as fissures or inflammatory or allergic processes or atopic dermatitis. It is also an object of the present invention to provide stable skin-care cosmetic and / or dermatological compositions which protect the skin from environmental influences how to protect sun and wind. In particular, the effect of the preparations should be fast and sustainable.

AQP stimulators can act in different ways. According to preferred A-quaporin stimulators enhance the expression of aquaporin AQP3, AQP5, AQP7 and AQP9, especially in AQP3 large increases are possible with the preparations according to the invention.

By quantitating the mRNA for AQP-3 and Western blotting, it can be demonstrated that the number of aquaporins in the epidermis increases significantly by application of aquaporin stimulants containing preparations according to the invention.

In Western blotting, the proteins from lysates of the skin in gels are electrophoretically separated by molecular weight and then transferred to a nitrocellulose membrane and thereby immobilized. By incubating the proteins on the membrane in an antibody solution specific for AQP, the AQP is selectively labeled and can be qualitatively and quantitatively detected by means of downstream detection and staining steps.

When quantifying the mRNA level of a protein, the copy number of the genetic material portion (DNA) for the protein in a cell is determined. The mRNA copies serve as a template for the synthesis of the protein at the cellular level and are directly preceded as a quantifiable size of the finished protein.

According to the invention Aquaporinstimulatoren from the group of

Glyceryl glycosides, in particular hexosylglycerides and / or (hexosyl) hexosylglycerides

CAMP analogs,

• PKA (adenylyl cyclase) activators and

• phosphodiesterase inhibitors, especially caffeine, theophylline

selected.

According to the use according to the invention, the cosmetic and dermatological preparations are characterized in that the cosmetically or pharmaceutically harmless aquaporin modulators are present in concentrations of 0.0001 to 20.00% by weight, preferably 0.0005-10.00% by weight, particularly preferably 0.001-5.00% by weight, in each case based on the total weight of the composition, is present or present.

Particularly preferred are modulators for the formation of aquaporins AQP3 and AQP5 to choose.

Particularly advantageous according to the invention are the D- and / or L-hexosylglycerides which induce the formation of new aquaporin-3 proteins. They activate the protein kinases contained in the cells, in particular protein kinase A, which stimulates aquaporin expression. Experiments on cell cultures have shown that addition of aquaporin stimulators according to the invention to the culture medium can lead to a tripling of the AQP-3 number (see Example 1).

Mitogen-activated kinases (Galcheva-Gorgova et al., Science 1994 (catalysed by glycosylglycerides) are then able to phosphorylate and thus in turn activate certain serine and threonine sites on many other intracellular proteins It also includes some of the transcription factors needed to make mRNA copies of the genetic material strand, and these activated transcription factors can then penetrate into the nucleus, causing mRNA copies of the gene segment - here: AQP-3 - and more aquaporin-3 produced in protein form in the cell.

Glyceryl glycoside (glucosylglyceride) is preferred for the stimulation of the aquaporin expression.

The hexoses underlying the hexosylglycerides used according to the invention are preferably selected from the group of aldohexoses, usually in their pyranoid form, ie allo (pyrano) s, altro (pyrano) s, gluco (pyrano) s, manno (pyrano) s, gulo (pyrano ), ldo (pyrano) se, galacto (pyrano) se and talo (pyrano) se.

The (hexosyl) hexoses on which the (hexosyl) hexosylglycerides according to the invention are based can be selected from the group of pyranosylpyranoses and furanosylpyranoses having 1,4-glycosidic or 1,6-glycosidic bond. They are preferably selected from the group maltose, leucrose, lactose.

Accordingly, the hexosylglycerides according to the invention can be characterized by the general structural formulas

respectively.

and the (hexosyl) hexosylglycerides of the invention by the general structural formulas

respectively.

mark.

It is advantageous to use D-hexosylglycosides, although L-hexosylglycosides can also be used with high activity in the context of the present invention.

Also hexosylglycerides, which are based on D- or L-ketohexoses, so psicose, fructose, sorbose or tagatose, usually present in their furanoid form, may optionally be used advantageously in the context of the present invention.

Glucosylglycerides of the general formula

and / or the general formula

are preferred according to the invention.

Particularly preferred hexosylglyceride is D-glucosylglycerol.

In particular, it is advantageous if hexosylglycerides of natural origin are used.

It was unforeseeable for a person skilled in the art that the glycosylglycerides according to the invention or cosmetic or dermatological preparations containing them

act better as a moisturizing agent and better against aging skin

as the active ingredients, drug combinations and preparations of the prior art.

According to the invention, the preparations contain from 0.001 to 15% by weight of glycosylglycerides, in particular from 0.01 to 9.5% by weight, very particularly preferably from 0.1 to 5% by weight.

According to the invention, the cosmetic preparations, in addition to the Aquaporinstimulatoren also substances which cause an osmotic stress on the treated skin areas and thus achieve a further increase in moisture content, included.

According to the invention, advantageous substances for triggering osmotic stress are:

Inorganic salts (especially alkaline earth and alkali metal salts which have a chloride, Sufat, hydrogen sulfate, phosphate, hydrogen phosphate, linear and / or cyclic ON gophosphate, carbonate or bicarbonate anion., Especially NaCl, NaBr , NaI, Na ₂ B ₄ O ₇ , Na ₂ SiO ₃ , Na ₂ CO ₃ , NaHCO ₃ , Na ₃ PO ₄ , Na ₂ HPO ₄ , NaH ₂ PO ₄ , KCl, KI, LiCl, NH ₄ Cl, ZnCl ₂ , Al ₂ SO ₃ , MgCl and MgSO ₄ ) salts of naturally occurring skin acids (eg energy metabolism such as sodium liponate, sodium citrate, ammonium lactate, sodium lactate, sodium bicarbonate, sodium citrate) or weak carboxylic acids (e.g. Sodium propionate) Natural mixtures of salts, in particular sea minerals, sugars with a molecular weight of up to 600 g / mol, in particular sorbitol, mannitol, sucrose, glucoses Amino acids, in particular glycine, alanine and / or asparagine.

It is advantageous to formulate the cosmetic preparations in such a way that, in addition to a water and / or oil phase, further cosmetically advantageous constituents are contained. In particular, antioxidants, moisturizers, sunscreen filters, anti-inflammatory agents and pigments have additional synergistic effects.

Example 1: Effect of glyceryl glycosides on AQP-3 expression

Figure 1 shows the level of expression of aquaporin-3 mRNA in human keratinocytes relative to a constitutive (non-modulatable) expressed gene, the 18S rRNA.

Glyceryl glucoside has a better stimulating effect than the classic glycerol or glucose alone or a 1: 1 mixture of glycerol and glyceryl glucoside.

For this purpose, human keratinocytes in cell culture (37 ° C., Medium Cambrex No. CC-3158, including Supplement Kit No. CC-4152, + 0.1 mM CaCl ₂ ) were treated for 24 h in triplicates as follows:

- untreated control (corresponds to 330mOsm, isosmolar culture medium)

- Addition of 1% w / v glycerol to the culture medium (corresponds to 450mOsm, osmotic stress) - Addition of 1, 5% w / v glycerol glucoside and 0.5% w / v glycerol to the culture medium

(corresponding to 450 mOsm osmotic stress, mixing ratio with respect to particle number 1: 1) addition of 3% w / v of glycerol glucoside to the culture medium (corresponds to 450 mOsm, osmotic stress)

Addition of 2.25% w / v glucose to the culture medium (corresponds to 450 mOsm, osmotic stress)

The different amounts used are due to the different molecular weight of the substances. Because the contribution to Osmolaritätserhöhung in the culture medium is only dependent on the added number of particles, and this is the same in all experiments.

After harvesting and lysis, the total RNA was isolated from the cells and the relatively contained aquaporin-3 mRNA was determined by quantitative RT-PCR.

The concentration w / v means mass per volume, where 1, 0% w / v corresponds to one gram of substance in 100 ml of solution.

Claims

claims

1. Use of glyceryl glycosides and / or derivatives thereof for stimulating the aquaporin expression.

2. Use according to claim 1 for stimulating the expression of AQP-3.

3. Use according to claim 1 for stimulating the expression of AQP-5 and / or AQP-7.

4. Use of cosmetic preparations containing one or more grylceryl glycoside and / or dreren derivatives to improve the transport of water and / or Moisturi- transport into the skin.

5. Use according to claim 5 for improving the Glycerintransportes in the skin.

6. Use according to at least one of claims 4 to 5, characterized in that the cosmetic preparation contains at least one substance which triggers the osmotic stress, wherein this is selected from the group:

Inorganic salts (in particular alkaline earth and alkali metal salts which have a chloride, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, linear and / or cyclic oligophosphate, carbonate or bicarbonate anion, especially NaCl, NaBr, NaI, Na ₂ B ₄ O ₇ , Na ₂ SiO ₃ , Na ₂ CO ₃ , NaHCO ₃ , Na ₃ PO ₄ , Na ₂ HPO ₄ , NaH ₂ PO ₄ , KCl, KI, LiCl, NH ₄ Cl, ZnCl ₂ , Al ₂ SO ₃ , MgCl and MgSO ₄ ),

Salts of acids naturally occurring in the skin, in particular salts of energy metabolism, such as sodium liponate, sodium citrate, ammonium lactate, sodium lactate, sodium bicarbonate, sodium citrate, weak carboxylic acids, sodium propionate,

Natural mixtures of salts, especially marine minerals, Sugar with a molecular weight of up to 600 g / mol, in particular glyceryl glucose, sorbitol, mannitol, sucrose, glucose and / or

Amino acids, in particular glycine, alanine and / or asparagine.

7. Use according to at least one of claims 4 to 6 for strengthening the barrier function of the skin.

8. Use according to at least one of claims 4 to 7 for improving the water and / or Moisturizertransport from the deeper layers of the skin (Stratum

Basal, the stratum spinosum or the stratum granulosum) to the skin surface and / or into the stratum corneum.