DE102011119953A1 - Individualized skin treatment comprises e.g. applying and removing adhesive tape on skin surface, obtaining adherent biological material, determining expression and/or activity of differentiation marker, and treating with cosmetic products - Google Patents

Abstract

Individualized skin treatment comprises (a) applying an adhesive tape on a selected skin surface, (b) removing the adhesive tape from the skin, (c) obtaining adherent biological material, (d) determining the presence, amount, expression and/or activity of at least one differentiation marker comprising filaggrin, transglutaminase and involucrin, (e) comparing the actual value with predetermined reference values, and (f) treating the skin of the individual with cosmetic products, which affect the presence, amount, expression and/or activity of at least one differentiation marker. Independent claims are also included for: (1) effectiveness evaluation of skin cosmetics, comprising (i) measuring the presence, amount, expression and/or activity of at least one differentiation marker comprising filaggrin, transglutaminase and involucrin of untreated skin area in an in vivo analysis, (ii) treating the skin area cosmetically with a topically applied skin cosmetic, (iii) measuring the presence, amount, expression and/or activity of at least one differentiation marker comprising filaggrin, transglutaminase and involucrin of the treated skin area in an in vivo analysis, (iv) comparing the value from step (i) with the value of step (iii), and (v) evaluating the effectiveness of skin cosmetic based on the influence of the presence, amount, expression and/or activity of the differentiation marker comprising filaggrin, transglutaminase and involucrin; and (2) use of tape stripping method for supporting advertising messages for cosmetics.

Description

The invention relates to a method for individualized skin treatment, which allows a targeted cosmetic treatment.

Well-groomed, smooth and firm skin is a consumer need across all cultures and age ranges. An almost infinite variety of skin cosmetic products wants to satisfy this consumer need, but this diversity also leads to uncertainty among consumers, which product is best suited for his skin and his desired appearance.

There is therefore a need to enable the consumer, beyond marketing promises, to select a product specially adapted to his skin. So far, research methods on the individual are reserved for medical research because they are either too time-consuming and therefore too expensive or require too much sample substance. An individual, reliable and also beyond medicine applicable method to judge the skin of a consumer and to offer targeted products based on the assessment does not exist in the cosmetic mass market.

In addition, consumers often want a "younger" look. However, corresponding statements in product advertising are perceived with skepticism.

The present invention was therefore based on the object to provide a method that allows a cost-effective and individual assessment of the skin condition and can make proposals for products depending on the assessment. The procedure should be feasible for the customer quickly and without significant impairments (pain, great expenditure of time).

• a) In Kontakt bringen eines Lebewesens, bevorzugt eines Menschen, mit einem zu testenden Wirkstoffs und/oder einer zu testenden Formulierung
• b) Aufbringen eines Klebebandes auf eine ausgewählte Hautoberfläche
• c) Abreißen des Klebebandes von der Haut
• d) Gewinnung von anhaftendem biologischem Material
• e) Detektion der Anwesenheit, Menge, Expression und/oder Aktivität mindestens eines Markers für Hautalterung ausgewählt aus der Liste gemäß Anhang 1 oder Anhang 2 des Dokuments.

Non-invasive methods for the screening of active substances and / or formulations against skin aging in vivo are known, for example, from US Pat DE 10 2009 046 128 A1 known. The method disclosed therein comprises the steps

• a) bringing into contact a living being, preferably a human, with an active ingredient to be tested and / or a formulation to be tested
• b) applying an adhesive tape to a selected skin surface
• c) Tearing off the adhesive tape from the skin
• d) Obtaining adherent biological material
• e) detection of the presence, amount, expression and / or activity of at least one skin aging marker selected from the list in accordance with Annex 1 or Annex 2 of the document.

The focus of the tape stripping method disclosed herein is to provide screening methods for obtaining statistical evidence for the development of cosmetic products. Neither the individual aspects mentioned above nor certain markers correlating particularly well in correlation with the cosmetic success are mentioned there.

It has now been found that a quick and painless in vivo analysis and communication of results with product recommendations not only solves the above-mentioned objects, but is also capable of significantly improving customer loyalty.

• a) ein Klebeband auf eine ausgewählte Hautoberfläche aufbringt,
• b) das Klebeband von der Haut abreißt,
• c) anhaftendes biologisches Material gewinnt,
• d) die Anwesenheit, Menge, Expression und/oder Aktivität mindestens eines Differenzierungsmarkers aus der Gruppe Filaggrin, Transglutaminase, Involucrin bestimmt,
• e) den Ist-Wert der Größe mit vorgegebenen Sollwerten vergleicht und
• f) die Haut des Individuums mit kosmetischen Mitteln behandelt, die die Anwesenheit, Menge, Expression und/oder Aktivität mindestens eines Differenzierungsmarkers aus der Gruppe Filaggrin, Transglutaminase, Involucrin beeinflussen.

The present invention is in a first embodiment, a method for individualized skin treatment, wherein

• a) applying an adhesive tape to a selected skin surface,
• b) the tape breaks off the skin,
• c) adherent biological material wins,
• d) the presence, quantity, expression and / or activity of at least one differentiation marker from the group of filaggrin, transglutaminase, involucrin,
• e) compares the actual value of the size with predetermined target values and
• f) the skin of the individual is treated with cosmetic agents which influence the presence, quantity, expression and / or activity of at least one differentiation marker from the group of filaggrin, transglutaminase, involucrin.

• i) im Rahmen einer in-vivo-Analyse die Anwesenheit, Menge, Expression und/oder Aktivität mindestens eines Differenzierungsmarkers aus der Gruppe Filaggrin, Transglutaminase, Involucrin eines unbehandelten Hautareals mißt,
• ii) das Hautareal mit einem topisch applizierten Hautkosmetikum kosmetisch behandelt,
• iii) im Rahmen einer in-vivo-Analyse die Anwesenheit, Menge, Expression und/oder Aktivität mindestens eines Differenzierungsmarkers aus der Gruppe Filaggrin, Transglutaminase, Involucrin des nunmehr behandelten Hautareals mißt,
• iv) den Wert aus Schritt i) mit dem Werte aus Schritt iii) vergleicht und
• v) die Wirksamkeit des Hautkosmetikums anhand des Einflusses auf die Anwesenheit, Menge, Expression und/oder Aktivität mindestens eines Differenzierungsmarkers aus der Gruppe Filaggrin, Transglutaminase, Involucrin bewertet.

In a second embodiment, the present invention relates to a method for evaluating the efficacy of skin cosmetics in which:

• i) measures the presence, amount, expression and / or activity of at least one differentiation marker from the group of filaggrin, transglutaminase, involucrin of an untreated skin area as part of an in vivo analysis,
• ii) cosmetically treating the skin area with a topically applied skin cosmetic,
• iii) in the context of an in vivo analysis, the presence, amount, expression and / or activity of at least one differentiation marker from the group of filaggrin, transglutaminase, involucrin of the now treated skin area is measured,
• iv) comparing the value from step i) with the value from step iii) and
• v) evaluates the efficacy of the skin cosmetic on the basis of the influence on the presence, amount, expression and / or activity of at least one differentiation marker from the group of filaggrin, transglutaminase, involucrin.

According to the first step of the method according to the invention, an adhesive tape is applied to a applied selected skin surface and removed in the subsequent step of her (demolished).

In particular, areas of skin with low hairiness are preferably selected or the hair is removed before the beginning of step a) in order to avoid pain or "sneezing" when taking the sample. It is particularly preferred in step a) to select the skin area from the skin surface of the arms, the legs, in particular the thighs, preferably the lower leg, the back or the facial skin. Preferably, the adhesive tape is applied in step a) on the skin of the forearms, in particular on the inside, or on the facial skin.

In order to avoid measurement errors and to obtain more sound statements, it is preferable to perform the sampling by means of adhesive tape (hereinafter also referred to as "tape stripping") on the relevant skin area multiple times. One and the same adhesive tape can be pressed onto the skin several times and pulled off, but it is preferable to apply fresh adhesive strips several times in succession to the same skin area and tear it off. Methods of the present invention wherein steps a) and b) involve sequentially applying and tearing off multiple adhesive tapes onto the same selected skin surface, preferably making at least 20 breaks, the first 10 of which are discarded, are therefore preferred.

The discarding of the first ten breaks serves to remove dirt, skin fat, dead skin cells, etc. as well as the removal of at least the uppermost layers of the stratum corneum. The following steps then represent "cleaner" samples, from which one can measure the presence, amount, expression and / or activity of at least one differentiation marker from the group filaggrin, transglutaminase, involucrin particularly well.

Filaggrin is a group of epidermal proteins that are formed in the process of keratinization of the skin. Filaggrins are also responsible for the cross-linking of keratin filaments via disulfide bridges in the stratum granulosum and in the stratum corneum and thus play an important role in the formation of an intact barrier. Mutations in the coding gene can lead to abnormally dry skin, hereditary forms of ichthyosis vulgaris, as well as susceptibility to atopic dermatitis type 2 with risk of allergies. In the course of progressive differentiation in the stratum granulosum, each of 10-12 filaggrin monomers forms the high molecular weight profilaggrin and deposits it in keratohyalin granules. At the border between stratum granulosum and stratum corneum, the proteolytic degradation of the profilaggrin then takes place to filagin molecules, which in turn form aggregates with the Keratinintermediärfilamenten. These aggregates can be further degraded to free amino acids, which then contribute to the Natural Moisturizing Factor of the skin. The Natural Moisturizing Factor may become a. Made from free amino acids, lactic acid, urea and various salts, it is also responsible for the ability of the skin to retain and store water. Filaggrin thus plays an essential role not only for the formation of an intact barrier but also for the maintenance of skin moisture.

Involucrin is formed by keratinocytes in the stratum spinosum keratinocytes and acts as a soluble precursor of the cornified envelope (CE) formed in the outermost layers. Invulcrin is cross-linked during terminal differentiation in the stratum granulosum by the enzyme transglutaminase and thus contributes to the formation of a stable Skin barrier at.

The skin forms a stable physical and chemical barrier against various environmental influences. The so-called "cornified envelope" (CE) thereby represents a critical structure for the barrier function of the outermost layer of the skin, the epidermis. The "cornified envelope" consists of an insoluble mix of various structural proteins such. B. involucrin or loricrin, which are covalently cross-linked during the terminal differentiation of the epidermal keratinocytes to a high molecular weight complex and so are essential to the formation of the barrier. Responsible for this cross-linking are transglutaminases, calcium-dependent enzymes, which catalyze the formation of an intermolecular isopeptide bond within or between the proteins polypeptide of participating proteins. Up to now, 8 transglutaminases have been identified in humans, with the transglutaminases 1, 3 and 5 in particular playing a role in the formation of the cornified envelope. A disruption of the transglutaminase activity can lead to a weakening of the skin barrier, so mice in which the coding gene for transglutaminase 1 has been knocked out show marked defects in the stratum corneum.

Further preferred methods according to the invention are characterized in that the non-discarded tears are pooled, wherein preferably ≥ 10, more preferably ≥ 12, even more preferably ≥ 15 and in particular ≥ 20 tears are evaluated together.

Preferably, several samples are pooled transferred to a sample tube and provided with buffer solution.

The recovery of the samples by means of the "tape-stripping method" enables the subsequent extracorporeal analysis of skin particles or cells. Since only one or a few skin particles or cells adhere to the adhesive tape, this process step is painless for the customer, can be carried out quickly and does not lead to an impairment of the external appearance. This step can be done by a beautician hairdresser, a salesman or even by the subject himself. The adhesive tape / tapes with the skin cells can then be subjected to the examination. This can be done in a simple handover, but it is also possible to transfer the samples in a suitable packaging and then bring to the examination site or to ship. In this way, sampling and examination can be decoupled spatially and temporally, even a transmission of samples by the customer itself can be done in this way.

• i. Ein- oder zweidimensionaler Gelelektrophorese
• ii. Affinitätschromatographie
• iii. Protein-Protein-Komplexierung in Lösung
• iv. ELISA (Enzyme-linked Immunosorbent Assay)
• v. enzymgekoppelte Stoffwechselassays
• vi. Massenspektrometrie, insbesondere Matrix Assistierter Laser Desorptions Ionisation (MALDI) und insbesondere
• vii. Einsatz von Proteinchips, Northern Blots,
• viii. Reverse Transkriptase Polymerasekettenreaktion (RT-PCR),
• ix. RNase-Schutzexperimente,
• x. Dot-Blots,
• xi. CDNA-Sequenzierung,
• xii. Klon-Hybridisierung,
• xiii. Differential Display,
• xiv. Subtraktive Hybridisierung,
• xv. cDNA-Fragment-Fingerprinting,
• xvi. Durchflusszytometrieanalysen,
• xvii. Serielle Analyse der Genexpression (SAGE) und insbesondere
• xviii. Einsatz von Nukleinsäurechips, oder mittels geeigneter Kombinationen dieser Methoden.

In a further step of the method according to the invention, the sample is examined for the presence, amount, expression and / or activity of at least one differentiation marker from the group filaggrin, transglutaminase, involucrin. In accordance with the invention preferred method, the investigation is carried out by a method which is selected from

• i. One- or two-dimensional gel electrophoresis
• ii. affinity
• iii. Protein-protein complexation in solution
• iv. ELISA (enzyme-linked immunosorbent assay)
• v. enzyme-linked metabolic assays
• vi. Mass spectrometry, in particular Matrix Assisted Laser Desorption Ionization (MALDI) and in particular
• vii. Use of protein chips, Northern blots,
• viii. Reverse transcriptase polymerase chain reaction (RT-PCR),
• ix. RNase protection experiments
• x. Dot blots,
• xi. CDNA sequencing,
• xii. Clone hybridization,
• xiii. Differential display,
• xiv. Subtractive hybridization,
• xv. cDNA fragment fingerprinting,
• xvi. flow cytometry analysis,
• xvii. Serial analysis of gene expression (SAGE) and in particular
• xviii. Use of nucleic acid chips, or by means of suitable combinations of these methods.

In particularly preferred methods according to the invention, the presence, amount, expression and / or activity of at least two differentiation markers from the group filaggrin, transglutaminase, involucrin are measured in step a) or i) and iii).

Such preferred methods measure the presence, amount, expression and / or activity of filaggrin and transglutaminase or of filaggrin and involucrin or of transglutaminase and involucrin.

In still further preferred methods of the invention, the presence, amount, expression and / or activity of all three differentiation markers filaggrin, transglutaminase, involucrin is measured in step a) or i) and iii).

In all cases mentioned above and regardless of which and how many differentiation markers are determined, the actual value of at least one differentiation marker is compared with predetermined desired values in step e) of the method according to the invention.

The accuracy can be further increased by measuring different skin areas and averages are formed from the measurement results thus determined.

These target values can be determined, for example, by the examination of healthy skin, wherein preferably mean values are formed from a collective. Particularly preferably, the collectives are formed in a particularly uniform and particularly large way, ie. H. the "Underlying" is determined from the skin of as many individuals as possible, sharing as many characteristics as possible, with age and gender being of the greatest importance. In preferred methods according to the invention, the elevation of the setpoint values in step e) is carried out with a sufficiently large (preferably at least 10, more preferably at least 50 and especially at least 100) control group with respect to age and gender.

Most particularly, further specifications / groupings are introduced and the limits set as narrow as possible. Thus, a control group "female, third decade of life" could be further divided into "female, 25 years" or "female, year of birth xxxx" and possibly even further in "female, 25 years, dry skin" etc.

In this way one obtains set values of certain collectives and can set the actual values in relation to it.

The comparison of the two measurements in steps i) and iii) of the second variant of the method can also be formulated if an effect on the presence, amount, expression and / or activity of the differentiation markers filaggrin, transglutaminase, involucrin occurred after the cosmetic treatment is.

On this data basis, the cosmetic agent applied in step ii) of the method according to the invention can then be evaluated in step v). The customer can thus be given an individual product recommendation that specifically addresses problematic areas without the customer having to rely on general advertising claims such as "tighter" skin. In addition, the success of the respective cosmetic can be shown and documented by further analysis after a certain treatment time. The individual satisfaction of the consumer is thus provided with an objective analysis that is suitable for binding the customer to a manufacturing company and its products.

Last but not least, the inventive method provides valuable services in the development of new cosmetics, as it allows a fast and objectified performance assessment of skin cosmetics.

Another object of the present invention is the use of the tape stripping method to support advertising claims for cosmetics.

QUOTES INCLUDE IN THE DESCRIPTION

This list of the documents listed by the applicant has been generated automatically and is included solely for the better information of the reader. The list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions.

Cited patent literature

• DE 102009046128 A1 [0006]

Claims (7)

A method for individualized skin treatment, characterized in that a) applying an adhesive tape to a selected skin surface, b) tearing the adhesive tape from the skin, c) adhering biological material, d) the presence, amount, expression and / or activity of at least one E) compares the actual value of the large with predetermined target values; and f) treats the skin of the individual with cosmetic agents which determine the presence, amount, expression and / or activity of at least one differentiation marker of the group filaggrin, transglutaminase, involucrin. Process for evaluating the efficacy of skin cosmetics, characterized in that i) measures the presence, amount, expression and / or activity of at least one differentiation marker from the group of filaggrin, transglutaminase, involucrin of an untreated skin area as part of an in vivo analysis, ii) cosmetically treating the skin area with a topically applied skin cosmetic, iii) in the context of an in vivo analysis, the presence, amount, expression and / or activity of at least one differentiation marker from the group of filaggrin, transglutaminase, involucrin of the now treated skin area is measured, iv) comparing the value from step i) with the value from step iii) and v) evaluates the efficacy of the skin cosmetic on the basis of the influence on the presence, amount, expression and / or activity of at least one differentiation marker from the group of filaggrin, transglutaminase, involucrin. Method according to one of claims 1 or 2, characterized in that steps a) and b) comprise successively applying and tearing off several adhesive tapes onto the same selected skin surface, wherein preferably at least 20 tears are made, of which the first 10 are discarded. A method according to claim 3, characterized in that the non-discarded tears are pooled, wherein preferably ≥ 10, more preferably ≥ 12, even more preferably ≥ 15 and in particular ≥ 20 tears are evaluated together. Method according to one of claims 1 to 4, characterized in that in step a) or i) and iii) the presence, amount, expression and / or activity of at least two differentiation markers from the group filaggrin, transglutaminase, involucrin is measured. Method according to one of claims 1 to 5, characterized in that in step a) or i) and iii) the presence, amount, expression and / or activity of all three differentiation markers filaggrin, transglutaminase, involucrin is measured. Use of the tape stripping method to support promotional claims for cosmetics.