WO2017221161A1

WO2017221161A1 - Method for obtaining a custom cosmetic treatment

Info

Publication number: WO2017221161A1
Application number: PCT/IB2017/053682
Authority: WO
Inventors: Adriano ALDEGHERI
Original assignee: Dermagenomic S.R.L.
Priority date: 2016-06-23
Filing date: 2017-06-21
Publication date: 2017-12-28
Also published as: EP3474813A1; ITUA20164627A1

Abstract

A method for evaluating the skin health status and for obtaining a custom composition for cosmetic use is described comprising the steps of obtaining a genomic DNA sample from a body tissue, isolating said genomic DNA sample, using said DNA sample to evaluate the presence of single nucleotide polymorphisms (SNPs) and preparation of a composition for cosmetic use based on the presence or absence of SNPs on the selected genes.

Description

"Method for obtaining a custom cosmetic treatment"

* * * *

The present invention relates to a method for obtaining a custom cosmetic treatment.

Several factors can affect the well-being and appearance of body tissues, including genetics, diet, physical activity, stress, hormone levels, personal hygiene and UV exposure. Experts have identified for years some molecules that can play a key role in maintaining the well-being of our tissues, and in particular, of the skin. Among these molecules there are antioxidants, fatty acids specifically mixed with other active agents, various vitamins, mineral co-factors and botanical elements, such as herbs and phyto-therapeutics.

The activity of these molecules and elements is related to their ability to play a specific role or action in one or more biological pathways. This role includes the ability to interact with elements of our body and to modify some biological processes to obtain a beneficial response for our skin.

The ability of an active ingredient to act, i.e., to give an efficient response, can be affected by changes in the genetic pattern of an individual. In this sense, in recent decades, small genetic differences have been analyzed, leading to a different response from individual to individual to an active ingredient. Single-Nucleotide Polymorphisms (SNPs) are the most important and basic form of genome variation. They are responsible for individual differences in drug response or for different susceptibility to pathologies. The analysis of the presence of SNPs is used for pharmacogenomic studies, i.e., studies designed to evaluate the different efficacy of an active ingredient in the presence or absence of such genetic variations. Other SNPs have been identified to evaluate the susceptibility of a patient to develop a particular pathology.

Even the administration of active substances for cosmetic use has undergone remarkable transformations in recent years, in fact the obtaining of a custom treatment that can be of maximum efficiency is aimed. Presently, the cosmetic preparations are administered on the basis of the symptoms and not on the basis of the causes that lead to a particular change in the state of the skin. Customers themselves ask for a custom and proven effectiveness treatment.

Over the years, several studies have highlighted the importance of genetics in preventing and treating aesthetic defects.

In particular, US 2016/0068904 describes a method for analyzing genetic variants that affect the aesthetic appearance of individuals, in particular of the skin. Said method provides for the analysis of genetic variants, such as the SNPs of some important genes for maintaining the well-being of the skin. Said method provides for the analysis of said SNPs and the use of specific products to be used in the presence of a specific SNP.

In addition, US 2015/0105279 describes a method for customizing cosmetic or nutri-cosmetic treatments. Said method provides for a preventative genetic analysis adapted to help the choice of the active ingredients for skin care. The selected active ingredients will then be used to prepare a cosmetic, cosmeceutical or nutri-cosmetic formulation.

US 2015/0086104 describes a method for evaluating the skin quality and for the assigning of a score. Said score is evaluated against a maximum score for human skin quality and then the best product or compound for the cosmetic treatment is evaluated. Among the various evaluated factors there is the presence of SNPs in particular genes.

WO 2014/176425 describes a method for obtaining a custom cosmetic preparation. Said method provides for the analysis of different SNPs including SNP rsl059261 of MMP-1.

Jordi Naval et al., Clinic. Cosm. and Invest. (2014), describes a study for the analysis of the incidence of some SNPs on the skin phenotype of some patients, among the various SNPs, rsl059261 of MMP-1 is evaluated. WO 2016/005793 A 1 describes a method for the preparation of a custom cosmetic composition, said method provides for the study of HAS-1 gene, inter alia, and the evaluation of all the SNPs thereof.

US 2009/036545 Al describes the use of bakuchiol in the treatment of the skin.

US 2012/114719 Al describes the use of essential and non-essential amino acids, N-acetylcysteine and madecassoside for the formulation of a composition for cosmetic use.

Eunsun Jung et al., Molecules, (2013) describes the use of madecassoside for the formulation of a composition for cosmetic use.

All of these documents describe an attempt to customize a cosmetic treatment as much as possible. The great disadvantage of these methods is the fact that the assessed factors are considered individually. The genetic variants are evaluated only on the basis of a possible cause, but without a wider picture with a real finding of the health status of the cutaneous tissue.

In addition, said methods aim at obtaining a remedy for each detected SNP by recommending lists of active ingredients, which are already known for decades, in the treatment of aesthetic defects, without any evaluation of possible synergistic effects of said genetic variants.

The object of the present invention is to provide a method for evaluating the health status of an individual's tissues, in particular of the cutaneous tissue.

Another object is to obtain useful information for the treatment of an individual's skin through targeted genetic testing.

An additional object is to obtain a method for preparing a custom cosmetic treatment according to an individual's skin health status.

Another object is a method for evaluating genetic variants on responses to certain active ingredients or substances.

According to the invention, this object is achieved by a method as described in claim 1.

These and other features of the present invention will be made more apparent from the following detailed description in a practical embodiment thereof.

Terminology

In the present invention, scientific terms and definitions of molecular biology are used which, unless explicitly described, will have the conventionally accepted meaning in the international scientific community.

The DNA (deoxyribonucleic acid) is a long chain polymer that contains the genetic material; most living organisms have their own DNA. The DNA consists of four units that are repeated within the polymer, the so- called nucleotides, whose order determines a particular signal. Depending on the signal contained in the DNA, several RNA chains (ribonucleic acid) are transcribed from which proteins are thus obtained. The nucleotides in the DNA are: Guanine (G), Cytosine (C), Adenine (A) and Thymine (T). The order in which the nucleotides are arranged identifies a particular amino acid that will thus form the proteins.

"Gene" refers to a DNA segment that contains protein-encoding sequences, wherein the segment may include exons, introns, promoters or other non-transcribed regions.

In genetics, "alleles" are defined as the two or more alternative forms of the same gene which are located in the same position on each homologous chromosome (genetic locus). The alleles control the same character but can lead to quantitatively or qualitatively different products.

The term haplotype (haploos = single or simple) defines the combination of allelic variants along a chromosome or a chromosomic segment containing loci in linkage disequilibrium, i.e., closely associated with each other, and which are usually inherited together.

A genetic marker is a DNA portion with a known location on a chromosome and that can be used to identify individuals or species.

The "genetic predisposition" is the susceptibility of a subject to a particular condition or pathology. Detecting a genetic predisposition can mean detecting a susceptibility or a pathology. In addition, by detecting a predisposition, it is possible to have predictive indications about a possible development of an interesting pathology or medical condition.

A "polymorphism" is a variation of nucleotides within DNA that causes a change in the amino acid sequence of the protein.

A "single nucleotide polymorphism" (SNP) is a variation of a single nucleotide in the DNA. Usually in literature SNPs are found in gene coding regions, or in non-coding regions or in intergenic regions between genes. There may be SNPs that give rise to the same amino acid, therefore to the so-called silent mutations and they are called "synonymous", while if a different amino acid is obtained from the nucleotide variation, we will have a "non-synonymous" SNP.

The present invention discloses a method for skin analysis to identify the skin health status. This method provides for the identification of genetic variants that may affect the skin health status and lead to blemishes and dysfunctions. The method uses individual human genetic information to identify the skin condition and to administer the best cosmetic product.

In a particular aspect, the invention provides for the analysis of the possible relationship between the presence of nucleotides in sites that give rise to polymorphisms on genes associated with skin maintenance and alterations.

The complex aetiology associated with skin imperfections is influenced by both genetic and environmental factors unique to each situation. In addition, these imperfections are caused by innumerable environmental, physiological and behavioural dynamics, such as the natural aging process, diet, sex, smoking, physical activity, allergens, UV exposure, sleep, hormones, the use of alcohol and tobacco.

Assessing all these factors simultaneously would be very difficult. The method object of the present invention provides the evaluation of the most active or inactive metabolic pathways in an individual that regulate the skin maintenance.

Said method firstly involves the analysis of said metabolic pathways through the biological material isolated from the individual. Said biological sample can be obtained by bioptic or surgical removal of the tissue, said tissue can be not-fixed, frozen, fixed in formalin and/or incorporated in paraformaldehyde; it may be saliva, tears, skin scrapes or other body fluids.

The method therefore aims to obtain a custom genetic profile of each individual's skin. Said genetic profile can be obtained by the following process:

- Obtaining the DNA genomic sample, for example from a skin, saliva or other tissue sample;

Isolating the DNA from the tissue;

Verifying the DNA quality and concentration;

Amplifying DNA by PCR (polymerase chain reaction);

- Using DNA as a template to detect/amplify the SNP regions for sequence determination;

Sequencing the SNP regions to assess the presence of alleles/variants;

Generating a custom genetic profile;

- Customizing the preparation of cosmetic, nutri-cosmetic, cosmeceutical products.

The applicant evaluated a panel comprising many genes involved in the turnover and maintenance of the skin, by selecting genes and statistically more significant variants thereof. The analysis of said genes has allowed to specifically evaluate four groups of genes and divide them according to the process to which they are linked.

In particular, the applicant identified four situations involved in skin aging:

The loss of skin elasticity;

- The Photo-aging; The presence of skin spots;

The loss of skin tone with wrinkle formation.

Loss of skin elasticity

The extracellular matrix is crucial for the consistency and elasticity of the skin and the wrinkle formation depends on it. The oxygen radical species (henceforth ROS) are chemically reactive molecules that make numerous chemical compounds unstable. The main ROS targets are DNA, proteins and membrane phospholipids. The ROS in the skin cause damage to the production of collagen and elastin, the main components of the extracellular matrix that functions to support and nourish the skin. This ROS action is one of the main causes of skin aging. A contrast action against ROS is carried out by glutathione, an antioxidant molecule. One of the components of glutathione (or gamma-glutamyl-cysteinyl-glycine) is cysteine, whose bioavailable form is N-acetylcysteine (NAC).

The NAC is commonly used as mucolytic by oral or inhalation route, but other applications have also been described, such as intra-dermal administration thereof in combination with a solution of essential and nonessential amino acids (commonly used for parenteral nutrition). The effects of such intra-dermal administration have been studied on cultured fibroblasts, obtaining a complete picture of the gene response of the major extracellular matrix components to that biochemical stimulus.

In particular, the genes most involved in maintaining the extracellular matrix have been studied. It was found that several genes have been induced by the combined administration of NAC and amino acids, thus achieving an effective extracellular matrix remodelling effect.

The selected genes are:

- HAS 1 (Locus: 19ql3.41; MEVI (Mendelian Inherited in Man) 601463): hyaluronic acid synthetase. The HAS 1 up regulation leads to a higher cellular production of hyaluronic acid, causing a moisturizing, invigorating and volumetric restoration of the dermis.

- HYAL1 (Locus: 3p21.31; MIM 607071): hyaluronidase 1. The HYAL1 up regulation leads to an increase of hyaluronic acid catabolism, allowing a replacement of the extracellular matrix.

- ELANE (Locus: 19pl3.3; MIM 130130) elastase. The ELANE up regulation leads to an increase in elastin catabolism, allowing a turnover of the extracellular matrix.

- MMP2 (Locus: 16ql2.2; MIM120360) type A gelatinase. The MMP2 up regulation leads to an increase in the non-fibrillar collagen IV catabolism present in the basal membrane, allowing a replacement of the dermal composition.

- MMP3 (Locus: l lq22.2; MIM 185250) stromelysin. The MMP3 up regulation leads to an increase in the catabolism of extracellular matrix components such as proteoglycans, fibronectin, laminin and type IV collagen, allowing a replacement of the dermal composition.

- MMP13 (Locus: l lq22.2; MIM 600108) type III collagenase. The MMP13 up regulation leads to collagen I, II and III increased catabolism, allowing a replacement of the dermal composition.

The induction of these genes, following the injective stimulus, occurs in the case of absolute integrity of the nucleotide sequence (not mutations altering the translation of the protein) and in the presence of SNP rs61736495 [C>T (Fwd); exon 3; synonymous replacement S238S; MAF (T): 0.0088] of the HAS l gene that allows to target the use of NAC, as a booster, only in those subjects that have a cytosine (C) in that position.

Photo-aging

Photo-aging, i.e., the UV rays mediated skin aging, is caused by the damages caused by over-exposure to the ultraviolet rays of cellular structures. The photo-aged skin is characterized by a decrease in thickness and elasticity, dryness, alteration of the epidermal barrier and hyper- pigmentation. Many of the UV damages are mediated by ROS, that make many chemical compounds unstable and alter the connective tissue. The ROS action can be counteracted by antioxidant compounds.

A group of genes involved in the maintenance of the connective tissue has been evaluated, particularly those genes that adjust the turnover of the major components of said connective tissue, such as collagen.

From this group of genes, those genes that were hyper-expressed following a stimulation with an antioxidant agent have been selected. Following numerous experiments, the applicant selected a potent antioxidant that can counteract the action mediated by ROS, i.e. astaxanthin (henceforth

AX).

AX is a carotenoid present mainly in algae and crustaceans (such as salmon, lobster, shrimp, crab, etc.) and has a well-known antioxidant, antiinflammatory and immunomodulatory activity. The AX main mode of action is to prevent and block the action of ROS by inhibiting the UV induced oxidative damage in cells, such as the peroxidation of lipids constituting the cell membrane. AX is poorly water-soluble, therefore liposomal formulations of AX (AX-lipo) have been prepared that allow the transcutaneous administration thereof.

To enhance the effect of AX, it was administered in combination with hydrolysed collagen (henceforth IdroCo), involved in the extracellular matrix synthesis and used with benefit to slow down skin aging, mainly caused by photo-aging.

The effects of transdermal administration of AX-IdroCo have been studied both in vitro and in vivo obtaining a picture of the gene response of the major target genes.

In particular, it has been found that the following genes are hyper- expressed or suppressed by the AX-IdroCo combined administration, thus producing a powerful anti-photo-aging effect.

- COL1A1 (Locus: 17q21.33; MIM 120150): type 1 -alpha 1 collagen. Essential component of the extracellular skin matrix. The administration of AX-IdroCo induces the transcription of this gene by increasing the collagen production.

- FBN1 (locus: 15q21.1; MIM 134797): fibrillin. Essential component of the extracellular skin matrix. The AX-IdroCo administration induces the transcription of this gene by increasing the fibrillin production.

- HAS 1 (Locus: 8q24.13; MIM 601636): hyaluronic synthetase 2.

The AX-IdroCo administration induces the transcription of this gene by increasing its production and consequently the amount of the produced hyaluronic acid.

- MMP1 (Locus: l lq22.2; MIM 120353): collagenase. The AX- IdroCo administration suppresses the transcription of this gene by actually reducing the collagen degradation action.

- MMP12 (Locus l lq22.2; MIM 601046): elastase. Administration of AX-IdroCo suppresses the transcription of this gene by effectively reducing the elastin degradation action.

The induction or the repression of these genes following the AX-IdroCo administration occurs in the case of absolute integrity of the nucleotide sequence (not mutations altering the translation of the protein); moreover, the presence of SNP rs 1799750 [-/G (Fwd); promoter at 5' UTR; insertion of a guanine (G); MAF (-): 0.43] of the MMP1 gene allows to adjust the AX dosage as a booster only in those subjects that present nucleotide insertion in that position, since it, in itself, results in a doubling of gene transcription.

Skin spots

The melanogenesis is the biochemical process, carried out by melanocytes, leading to the formation of melanin. The melanogenesis is induced by UV rays and is a physiological process that protects epidermal cells from the attack of free radicals that cause DNA damage. However, this process accelerates skin aging and can give rise to unsightly, dark spots. The UVs indirectly influence the melanogenesis of melanocytes by a regulatory paracrine process involving keratinocytes.

In the epidermis, PEG2 and PGF2A are the major prostaglandins that constitute the mediators produced by keratinocytes in response to UVs. Several studies indicate that prostaglandins mediate post-inflammatory pigmentation changes by modulation of melanin and melanocytic dendrites synthesis.

The receptor activated by type 2 protease (PAR2) is expressed in keratinocytes but not in melanocytes, and plays a key role in the regulation of the cyclooxygenase 2 enzyme (COX-2), which together with the PEG2 enzyme (prostaglandin E2 synthase), produces the prostaglandins, the inflammation mediators. PAR2 increases the transcription of COX-2 and the release of arachidonic acid, a precursor of prostaglandins, causing increased secretion of PGE2 and PGF2. Finally, PAR2 regulates the uptake of melanosomes in keratinocytes.

The madecassoside (henceforth MA) is one of the major triterpene glycosides isolated from Centella Asiatica and has various skin effects, including an anti-inflammatory, anti-aging and protective effect against oxidative stress and UVB rays. The anti-inflammatory activity is due to the inhibition of COX-2 and PAR2 following exposure to UVB, thus reducing the production of prostaglandins. MA inhibits the phagocytosis of melanosomes, induces melanin synthesis, melanosome increase and melanocytic dendritogenesis. Finally, MA has been shown to be effective, in vitro, in increasing the production of type 1 and 3 collagens in fibroblasts. MA is poorly water-soluble, therefore MA liposomal formulations

(MALipo) that allow the trans-cutaneous administration thereof have been created.

The MA effects have been studied in vitro obtaining a picture of the gene response of the main target genes of this compound.

In particular, it has been found that the following genes are inhibited by MA administration thus achieving an effective skin anti-pigmentation effect.

- PAR2 (o F2RL1) (Locus 5Q13.3; MIM 600933): receptor activated by type 2 proteases. It regulates the COX-2 enzyme. The MA administration, in the presence of UVBs, inhibits the transcription of this gene, by reducing the production of PEG2 and

PGF2A.

- COX-2 (or PTGS2) (Locus lq31.1; MIM 600262): type 2 cyclooxygenase or prostaglandin synthase 2. It catalyses the first step of conversion of arachidonic acid into prostaglandins. The MA administration, in the presence of UVBs, inhibits the transcription of this gene, by inhibition of PAR2.

- COL1A1 (Locus 17q21.33; MIM 120150): 1-alpha type 1 collagen. Essential component of the extracellular skin cell matrix. The MA administration induces the transcription of this gene by increasing the collagen production.

- COL3A1 (Locus 2q32.2; MIM 120180): 3-alpha type 1 collagen.

Fibrillar essential component of the extracellular matrix of the skin. The MA administration induces the transcription of this gene by increasing the collagen production.

The induction or inhibition of the transcription of these genes following the MA administration occurs in the case of absolute integrity of the nucleotide sequence (not mutations that alter the translation of the protein); moreover, the presence of SNP rs689466 [A7G (Rew); promoter at 5' UTR; A>G; MAF (G); 0.21] and of SNP rs20417 [G/C (Rew); promoter at the 5' UTR; G>C; MAF (G): 0.19] of the COX-2 gene, which allows to combine a higher sunscreen at MA dosage, used as booster, only in those patients who exhibit an A-G haplotype at the two SNPs; in fact, such combination maintains the expression of COX-2 at normal levels and therefore these subjects have a lower basal prostaglandin production due to exposure to UVBs and tend to have greater skin hyperpigmentation. Loss of the skin tone with wrinkle formation

Retinoids are natural compounds that include vitamin A (retinol) and derivatives thereof, such as retinal and retinoic acid. Retinoic acid is considered the active form of vitamin A and is involved in different gene regulation pathways with effects on both proliferation and cell differentiation.

The retinol is therefore a fundamental substance for skin growth and differentiation, and is metabolised in retinoic acid in skin fibroblasts (wrinkles, skin hyperpigmentation, acne, psoriasis) and improves the appearance of the skin (increased consistency and skin tone, increased elasticity). Unfortunately, retinols have several side effects, including skin irritation, dryness, burning sensation, which limit the use thereof, for this reason retinol-like substances have been identified that, while retaining the specific effect of retinol, do not possess the retinol side effects.

For example, Bakuchiol (henceforth referred to as Bak) is a meroterpene phenol present in the Psoralea Corylifolio plant, and it is one of those retinol-like compounds that has shown retinol-like effects and is used as anti-oxidant, anti-wrinkle and anti-acne.

The effects of the Bak administration have been studied in vitro by obtaining a picture of the gene response of the main target genes of this retinol-like compound.

In particular, it has been found that the following genes are induced or suppressed by the Bak administration, thus achieving an effective anti- wrinkle effect with skin tone rebalancing.

- N6AMT1 (Locus 21q21.3; MIM 614553): N6 DNA adenine- specific methyltransferase 1. Bak down-regulates the expression of N6AMT1, reducing the retinol-like toxicity by decreasing the epigenetic modifications to DNA mediated by this enzyme.

- DHRS9 (Locus 2q31.3; MIM 612131): Retinol dehydrogenase.

This enzyme is involved in the conversion of retinol to retinal and then into retinoic acid. Bak induces this enzyme by increasing the conversion of retinols.

- LRAT (Locus 4q32.1; MIM 604863): Lecithin-retinol acetyltransferase. This enzyme mediates the retinol esterification reaction with long chain fatty acids, key step to retinol uptake and storage.

- CRBP1 (Locus 3q23; MIM 180260); cellular retinol binding- protein 1.

- CRBP2 (Locus 3q23; MIM 180280); cellular retinol binding- protein 2.

- CRBP4 (o RBP7) (Locus lp36.22; MIM 608604); cellular retinol binding-protein 4.

CRBPs are proteins involved in cellular internalization and retinol cellular transfer, retinol esterification, retinol esters hydrolysis, and oxidation from retinol to retinaldehyde (key passage in retinoic acid biosynthesis). The expression of these three CRBPs is induced by Bak, which thus increases the bioavailability of retinol at cellular level.

- HAS 3 (Locus 16q22.1; MIM 602428): hyaluronic synthase 3. The Bak administration induces the transcription of this gene, by increasing its production and consequently the amount of the produced hyaluronic acid.

The variation in the expression of these genes following the administration of Bak, occurs in the case of integrity of the nucleotide sequence (not mutations that alter the translation of the protein) and in the presence of SNP rs 1059261 [C>T (Fwd); synonymous replacement L306L;

MAF (C): 0.274] of the DHRS9 gene that allows the dosage of Bak, as a booster, to be adjusted in those subjects presenting the base C in that position, since it causes a low gene transcription as such.

Depending on the SNPs that will be detected during screening, several active ingredients will also be administered in combination. The entire cascade of genes that compose the studied pathways will be also evaluated, analyzing the involvement of the different genes in the signal activation cascade, and accurately selecting, and according to the above- mentioned information, the booster to be included in the composition for cosmetic use.

There is, therefore, not only the evaluation of the presence/absence of one or more SNPs but also the general evaluation of the activation of the various pathways and, therefore, the general metabolic state of that particular studied signal.

The genes are analyzed by whole gene sequencing.

The composition for cosmetic use may be a cream, an ointment, a water in oil emulsion, an oil in water emulsion or any other type of emulsion, gel, paste, cataplasm, medicated poultice, medicated foam or medicated plaster. Said cosmetic composition may contain in addition to the active ingredients at least one excipient.

Said at least one excipient may be a lipophilic excipient, such as an oleaginous base (petralatum, fats, silicones, isopropyl myristate, etc.), an absorbent base (lanolin, lanolin alcohols, lano-vaseline, etc.) or hydrophilic excipient such as for example ethylene oxide derivatives, polyethylene glycols, water, hydrocolloids such as starch, methylcellulose, carboxymethylcellulose, gelatin, carbopol, or any other excipient for cosmetic use.

Said composition for cosmetic use may also contain surfactants, preservatives, antioxidants, dyestuffs or fragrances, which may be selected according to the national regulations of the countries in which it will be marketed.

In a first embodiment, said method for obtaining a custom composition for cosmetic use provides for the analysis of the four panels of genetic variants that will be analyzed together. At the end of the analysis, substances for cosmetic use will be administered which can also be prepared in the form of preparation for cosmetic use. The metabolic status of the pathways involved in the four panels chosen for the study will be evaluated.

The presence of at least one of the following SNPs will be therefore investigated: SNP rs61736495 of HAS 1 gene for the loss of skin elasticity; the presence of SNP rs 1799750 on the MMP1 gene to evaluate the photo- aging; the presence of SNPs rs689466 and rs20417 on COX2 gene to evaluate the presence of skin spots and the presence of SNP rs 1059261 on DHRS9 gene to evaluate the loss of skin tone and the wrinkle formation.

The presence/absence of said SNPs is closely related to the metabolic status of the pathway of interest and may affect its proper activation, therefore all the genes involved in the specific metabolic pathway will also be studied.

The cosmetic composition will be prepared according to the indications detected from the analysis of the at least one SNP. The composition for cosmetic use comprises at least one of the following active ingredients:

a solution of essential and non-essential amino acids (L-isoleucine at a concentration by weight between 0.4% and 0.65%, L-leucine at a concentration by weight between 0.65% and 0.85%, L-lysine monochlorohydrate at a concentration by weight between 0.65% and 0.85%, L-methionine at a concentration by weight between 0.3% and 0.5%, L-phenylalanine at a concentration by weight between 0.3% and 0.5%, L-threonine at a concentration by weight between 0.25% and 0.40%, L-tryptophan at a concentration by weight between 0.05% and 0.2%, L-valine at a concentration by weight between 0.4% and 0.65%, L-alanine at a concentration by weight between 0.4% and 0.65%, L-arginine at a concentration by weight between 0.7% and 0.9%, L-histidine hydrochloride at a concentration by weight between 0.15% and 0.35%, L-proline at a concentration by weight between 0.8% and 1%, L-serine at a concentration by weight between 0.4% and 0.65%, L-cysteine hydrochloride monohydrate at a concentration by weight between 0.15% and 0.3%, glycine at a concentration by weight between 0.9% and 1.3%) dissolved in a physiological solution added with electrolytes. The amino acids are dissolved in the physiological solution at a concentration between 3 and 12% by weight, preferably between 5 and 10%, still more preferably between 7 and 9%.

N-acetylcysteine (NAC) to a final concentration between 5 and 50 mg/ml, preferably between 10 and 35 mg/ml, still more preferably between 15 and 25 mg/ml.

Astaxanthine (AX) at a concentration between 0.5% and 2.5% by weight at 8 or 16 micronM in combination with Hydrolyzed Collagen. Preferably, the AX will be contained in a liposomal preparation.

Madecassoside (MA) at a final concentration between 0.5% and 1.5%, more preferably between 0.8% and 1.25% by weight. Preferably the MA will be contained in a liposomal preparation. A sunscreen selected from a group comprising: Ν,Ν,Ν-Trimethyl- 4-(2-oxoborn-3-ylidenemethyl) anilinium methyl sulphate, Benzoic acid, 2-hydroxy,3,3,5-trimethylcyclohexyl ester/Homosalate, 2-Hydroxy-4-methoxybenzophenone, 2- Phenylbenzimidazole-5-sulphonic acid and its potassium, sodium and triethanolamine salts, 3,3'-(l,4-Phenylenedimethylene) bis (7,7-dimethyl-2-oxobicyclo-[2.2.1] hept- 1-ylmethanesulfonic acid) and its salts, l-(4-tert-Butylphenyl)-3-(4-methoxyphenyl) propane- 1,3-dione, alpha-(2-Oxoborn-3-ylidene)toluene-4-sulphonic acid and its salts, 2-Cyano-3,3-diphenyl acrylic acid, 2-ethylhexyl ester, Polymer of N-{(2 and 4)-[(2-oxoborn-3- ylidene)methyl]benzyl}acrylamide, 2-Ethylhexyl 4- methoxycinnamate/Octinoxate, Ethoxylated Ethyl-4-

Aminobenzoate, Isopentyl-4-methoxycinnamate/Amiloxate, 2,4,6- Trianilino-(p-carbo-2'-ethylhexyl-l '-oxy)-l,3,5-triazine, Phenol, 2- (2H-Benzotriazol-2-yl)-4-Methyl-6-(2-Methyl-3-(l,3,3,3- Tetramethyl-l-(Trimethylsilyl)Oxy)-Disiloxanyl)Propyl, Benzoic acid, 4,4-{ [6-[[[(l,l- dimethylethyl)arnino]carbonyl]phenyl]amino]-l,3-5-triazine-2,4- diyl]diimino}bis-, bis(2-ethylhexyl)ester/Iscotrizinol, 3-(4'- Methylbenzylidene)-dl-camphor/Enzacamene, 3-Benzylidene camphor, 2-Ethylhexyl salicylate/Octisalate), 2-Ethylhexyl 4- (dimethylamino)benzoate/Padimate O, 2-Hydroxy-4- methoxybenzophenone- 5 -sulfonic acid (Benzophenone-5) and its sodium salt/Sulisobenzone, 2-Hydroxy-4-methoxybenzophenone- 5-sulfonic acid (Benzophenone-5) and its sodium salt/Sulisobenzone, 2,2 '-Methylene bis(6-(2H-benzotriazol-2-yl)-4- (l,l,3,3-tetramethylbutyl)phenol)/Bisoctrizole, Sodium salt of 2,2'- bis( 1 ,4-phenylene)- lH-benzimidazole-4,6-disulfonic

acid/Bisdisulizole disodium, 2,2'-(6-(4-Methoxyphenyl)-l,3,5- triazine-2,4-diyl)bis(5-((2-ethylhexyl)oxy)phenol)/Bemotrizinol, Titanium dioxide, Benzoic acid, 2-[4-(diethylamino)-2- hydroxybenzoyl]-, hexylester, 1,3,5-Triazine, 2,4,6-tris(l,l '- biphenyl)-4-yl-, including as nanomaterial and other sunscreens approved for cosmetic use by the legislation of the country where the cosmetic composition will be marketed.

Bakuchiol at a concentration by weight between 2% and 15%, preferably between 4% and 12% by weight.

In a second embodiment, the genetic panel relating to the loss of skin elasticity will be evaluated and in particular the HAS 1 gene will be analyzed to investigate the presence of SNP rs61736495. Also, the genes HYALl, ELANE, MMP2, MMP3 and MMP13 will be studied. For subjects with cytosine (C) in rs61736495, a cosmetic composition will be prepared comprising a solution of essential and non-essential amino acids (L- isoleucine at a concentration by weight between 0.4% and 0.65%, L-leucine at a concentration by weight between 0.65% and 0.85 %, L-lysine monochlorohydrate at a concentration by weight between 0.65% and 0.85%, L-methionine at a concentration by weight between 0.3% and 0.5%, L- phenylalanine at a concentration by weight between 0.3% and 0.5%, L- threonine at a concentration by weight between 0.25% and 0.40%, L- tryptophan at a concentration by weight between 0.05 % and 0.2%, L-valine at a concentration by weight between 0.4% and 0.65%, L-alanine at a concentration by weight between 0.4% and 0.65%, L-arginine at a concentration by weight between 0.7% and 0.9%, L-histidine hydrochloride at a concentration by weight between 0.15% and 0.35%, L-proline at a concentration by weight between 0.8% and 1%, L-serine at a concentration by weight between 0.4% and 0.65%, chloro -hydrochloride monohydrate L- cysteine at a concentration by weight between 0.15% and 0.3%, glycine at a concentration by weight between 0.9% and 1.3%) dissolved in a physiological solution added with electrolytes and N-acetylcysteine (NAC).

The amino acids are dissolved in the physiological solution at a concentration between 3 and 12% by weight, preferably between 5 and 10%, still more preferably between 7 and 9%.

NAC will have a final concentration between 5 and 50 mg/ml, preferably between 10 and 35 mg/ml, still more preferably between 15 and 25 mg/ml.

In a third embodiment, the genetic panel regarding the loss of skin elasticity will be evaluated and in particular the HAS 1 gene will be analyzed to investigate the presence of SNP rs61736495. Also, the genes HYAL1, ELANE, MMP2, MMP3 and MMP13 will be studied. For subjects with thymine (T) in rs61736495, a cosmetic composition will be prepared comprising a solution of essential and non-essential amino acids (L- isoleucine at a concentration by weight between 0.4% and 0.65%, L-leucine at a concentration by weight between 0.65% and 0.85%, L-lysine monochlorohydrate at a concentration by weight between 0.65% and 0.85%, L-methionine at a concentration by weight between 0.3% and 0.5%, L- phenylalanine at a concentration by weight between 0.3% and 0.5%, L- threonine at a concentration by weight between 0.25% and 0.40%, L- tryptophan at a concentration by weight between 0.05% and 0.2%, L-valine at a concentration by weight between 0.4% and 0.65%, L-alanine at a concentration by weight between 0.4% and 0.65%, L-arginine at a concentration by weight between 0.7% and 0.9%, L-histidine hydrochloride at a concentration by weight between 0.15% and 0.35%, L-proline at a concentration by weight between 0.8% and 1%, L-serine at a concentration by weight between 0.4% and 0.65%, L-cysteine hydrochloride monohydrate at a concentration by weight between 0.15% and 0.3%, glycine at a concentration by weight between 0.9% and 1.3%) dissolved in a physiological solution added with electrolytes.

In a fourth embodiment, the genetic panel relating to the photo-aging will be evaluated and in particular the MMP1 gene will be examined to investigate the presence of SNP rsl799750. COL1A1, FBN1, HAS2, MMP1 and MMP12 genes will also be studied. For subjects with only one guanine in rs 1799750 a cosmetic composition will be prepared comprising 8 microM liposomal AX at a concentration between 0.5 and 1.2% by weight. Preferably, said AX is associated with hydrolysed collagen (IdroCo).

In a fifth embodiment, the genetic panel relating to the photo-aging will be evaluated and in particular the MMP1 gene will be examined to investigate the presence of SNP rsl799750. COL1A1, FBN1, HAS2, MMP1 and MMP12 genes will also be studied. For subjects with two Guanines in rs 1799750, a cosmetic composition will be prepared comprising a 16 microM liposomal AX at a concentration between 1.4% and 2.5% by weight, preferably between 0.1 and 0.25% by weight. Preferably, said AX is associated with hydrolysed collagen (IdroCo).

In a sixth embodiment, the genetic panel relating to the presence of skin spots will be evaluated and in particular the COX2 gene will be examined to investigate the presence of SNPs rs689466 and rs20417. The PAR2, COX-2, COL1A1 and COL3A1 genes will also be studied.

For the subjects with an A-G haplotype in rs689466-rs20417, a cosmetic composition comprising liposomal MA will be prepared at a concentration between 0.5 and 1.5% by weight, preferably between 0.8 and 1.25% by weight. Preferably said MA is associated with a sunscreen with Solar Protection Factor (SPF) between 30 and 50.

For the subjects with a G-C haplotype in rs689466-rs20417, a cosmetic composition comprising liposomal MA will be prepared at a concentration between 0.5 and 1.5% by weight, preferably between 0.8 and 1.25% by weight. Preferably said MA is associated with a sunscreen with SPF between 6 and 25.

In a seventh embodiment, the genetic panel relating to the loss of skin tone and wrinkle formation will be evaluated and in particular the DHRS9 gene will be analyzed to investigate the presence of SNP rs 1059261. The genes N6AMT1, DHRS9, LRAT, CRBPl, CRBP2 and CRBP4 will also be studied.

For the subjects presenting the T in rs 1059261 a cosmetic composition comprising Bak at a concentration between 2% and 10% by weight, preferably between 4% and 8% by weight, will be prepared.

In an eighth embodiment, the genetic panel relating to the loss of the skin tone and wrinkle formation will be evaluated and in particular the DHRS9 gene will be analyzed to investigate the presence of SNP rs 1059261. The genes N6AMT1, DHRS9, LRAT, CRBPl, CRBP2 and CRBP4 will also be studied.

For the subjects presenting the C in rs 1059261 a cosmetic composition comprising Bak at a concentration from 4 to 13% by weight, preferably from 6 to 12% by weight, will be prepared.

Advantageously, said method allows a custom preparation of the cosmetic composition, dosing in a specific manner the amounts of active ingredients to be added to each different type of cosmetic composition.

Another advantage is that this method allows to analyze several aspects of a tissue and to evaluate the overall health status.

Another advantage is that this method provides for the analysis of selected ad hoc SNPs allowing a targeted tissue health analysis avoiding time and money waste.

Claims

1. A method for obtaining a custom cosmetic treatment comprising the steps of

obtaining a sample of genomic DNA from a body tissue,

isolating said genomic DNA sample,

using said DNA sample to evaluate the activation state of metabolic pathways by the study of selected genes and the presence or absence of single nucleotide polymorphisms (SNPs) and

administering one or more substances for cosmetic use according to the presence or absence of SNPs in the analyzed biological sample,

characterized in that

the genes are selected from a group comprising: N6AMT1, DHRS9, LRAT, CRBP1, CRBP2, CRBP4, COL1A1, FBN1, HAS2, MMP1, MMP12, HAS 1, HYAL1, ELANE, MMP2, MMP3, MMP13, PAR2, COX-2 and COL3Al,

the presence or absence of SNPs is investigated on a least one gene selected from a group comprising HAS 1, MMP1, DHRS9 and COX2,

the substances administered for cosmetic use are chosen from a group comprising: N-acetylcysteine, astaxanthin, madecassoside, a sunscreen, bakuchiol and a solution of essential and non-essential amino acids dissolved in a physiological solution added with electrolytes.

2. A method according to claim 1, characterized in that the SNPs are rs61736495, rsl799750, rsl059261, rs689466 and rs20417.

3. A method according to claim 1 or 2, characterized in that the genes analyzed are HAS 1, HYAL1, ELANE, MMP2, MMP3 and MMP13, the

SNP is rs61736495 on HAS 1 gene and the substances administered for cosmetic use comprise essential and non-essential amino acids dissolved in a physiological solution added with electrolytes and N-acetylcysteine, said essential and non-essential amino acids at a concentration by weight between 5% and 10% and said N-acetylcysteine at a final concentration between 10 and 35 mg/ml, said essential and non-essential amino acids comprising L-isoleucine at a concentration by weight between 0.4% and 0.65%, L-leucine at a concentration by weight between 0.65% and 0.85%, L-lysine monochlorohydrate at a concentration by weight between 0.65% and 0.85%, L-methionine at a concentration by weight between 0.3% and

0.5%, L-phenylalanine at a concentration by weight between 0.3% and 0.5%, L-threonine at a concentration by weight between 0.25% and 0.40%, L-tryptophan at a concentration by weight between 0.05 % and 0.2%, L- valine at a concentration by weight between 0.4% and 0.65%, L-alanine at a concentration by weight between 0.4% and 0.65%, L-arginine at a concentration by weight between 0.7% and 0.9%, L-histidine hydrochloride at a concentration by weight between 0.15% and 0.35%, L-proline at a concentration by weight between 0.8% and 1%, L-serine at a concentration by weight between 0.4% and 0.65%, L-cysteine hydrochloride monohydrate at a concentration by weight between 0.15% and 0.3%, glycine at a concentration by weight between 0.9% and 1.3%.

4. A method according to claim 1 or 2 characterized in that the genes analyzed are HAS 1, HYAL1, ELANE, MMP2, MMP3 and MMP13, the SNP is rs61736495 on HAS 1 gene and the substances administered for cosmetic use include essential and non-essential amino acids dissolved in a physiological solution added with electrolytes, said essential and nonessential amino acids at a concentration by weight between 5% and 10%, said solution of essential and non-essential amino acids comprising L- isoleucine at a concentration by weight between 0.4% and 0.65%, L-leucine at a concentration by weight between 0.65% and 0.85%, L-lysine monochlorohydrate at a concentration by weight between 0.65% and 0.85%, L-methionine at a concentration by weight between 0.3% and 0.5%, L- phenylalanine at a concentration by weight between 0.3% and 0.5%, L- threonine at a concentration by weight between 0.25% and 0.40%, L- tryptophan at a concentration by weight between 0.05% and 0.2%, L-valine at a concentration by weight between 0.4% and 0.65%, L-alanine at a concentration by weight between 0.4% and 0.65%, L-arginine at a concentration by weight between 0.7% and 0.9%, L-histidine hydrochloride at a concentration by weight between 0.15% and 0.35%, L-proline at a concentration by weight between 0.8% and 1%, L-serine at a concentration by weight between 0.4% and 0.65%, L-cysteine hydrochloride monohydrate at a concentration by weight between 0.15% and 0.3%, glycine at a concentration by weight between 0.9% and 1.3%.

5. A method according to claim 1 or 2, characterized in that the genes analyzed are COL1A1, FBN1, HAS 1, MMPl and MMPl 2, the SNP is rs 1799750 on MMPl gene and the substances administered for cosmetic use comprise astaxanthin 8 μιη at a concentration between 0.05% and 2.5% by weight in combination with hydrolyzed collagen.

6. A method according to claim 1 or 2, characterized in that the genes analyzed are COL1A1, FBN1, HAS 1, MMPl and MMPl 2, the SNP is rs 1799750 on MMPl gene and the substances administrated for cosmetic use comprise astaxanthin 16 μιη at a concentration between 1.4% e 2.5% by weight in combination with hydrolyzed collagen.

7. A method according to claim 1 or 2, characterized in that the genes analyzed are PAR2, COX-2, COL1A1 and COL3A1, the SNPs are rs689466 and rs20417 on COX2 gene and the substances administered for cosmetic use comprise madecassoside at a final concentration between 0,5% and 1,5% by weight associated with a sunscreen with SPF between 30 and 50.

8. A method according to claim 1 or 2, characterized in that the genes analyzed are PAR2, COX-2, COL1A1 and COL3A1, the SNPs are rs689466 and rs20417 on COX2 gene and the substances administered for cosmetic use comprise madecassoside at a final concentration between 0.5% and 1.5% by weight associated with a sunscreen with SPF between 6 and 25.

9. A method according to claim 1 or 2, characterized in that the genes analyzed are N6AMT1, DHRS9, LRAT, CRBP1, CRBP2 and CRBP4, the SNP is rs 1059261 on DHRS9 gene and the substances administered for cosmetic use comprise bakuchiol at a concentration between 2% and 10% by weight.

10. A method according to claim 1 or 2, characterized in that the genes analyzed are N6AMT1, DHRS9, LRAT, CRBP1, CRBP2 and CRBP4, the SNP is rs 1059261 on DHRS9 gene and the substances administered for cosmetic use comprise bakuchiol at a concentration between 4% and 13% by weight.

11. A method according to any one of the preceding claims, characterized in that the sunscreen is selected from a group comprising:

N,N,N-Trimethyl-4-(2-oxoborn-3-ylidenemethyl) anilinium methyl sulphate, Benzoic acid, 2-hydroxy,3,3,5-trimethylcyclohexyl ester/Homosalate, 2-Hydroxy-4-methoxybenzophenone, 2-

Phenylbenzimidazole-5-sulphonic acid and its potassium, sodium and triethanolamine salts, 3,3 '-(l,4-Phenylenedimethylene) bis (7,7-dimethyl-2- oxobicyclo-[2.2.1] hept-l-ylmethanesulfonic acid) and its salts, l-(4-tert- Butylphenyl)-3-(4-methoxyphenyl) propane- 1, 3 -dione, alpha-(2-Oxoborn-3- ylidene)toluene-4-sulphonic acid and its salts, 2-Cyano-3,3-diphenyl acrylic acid, 2-ethylhexyl ester, Polymer of N-{ (2 and 4)-[(2-oxoborn-3- ylidene)methyl]benzyl}acrylamide, 2-Ethylhexyl 4- methoxycinnamate/Octinoxate, Ethoxylated Ethyl-4-Aminobenzoate, Isopentyl-4-methoxycinnamate/Amiloxate, 2,4,6-Trianilino-(p-carbo-2'- ethylhexyl- 1 '-oxy)- 1 ,3,5-triazine, Phenol, 2-(2H-Benzotriazol-2-yl)-4- Methyl-6-(2-Methyl-3-(l,3,3,3-Tetramethyl-l-(Trimethylsilyl)Oxy)- Disiloxanyl)Propyl, Benzoic acid, 4,4-{ [6-[[[(l,l- dimethylethyl)amino]carbonyl]phenyl]amino]-l,3-5-triazine-2,4- diyl]diimino}bis-, bis(2-ethylhexyl)ester/Iscotrizinol, 3-(4'-

Methylbenzylidene)-dl-camphor/Enzacamene, 3-Benzylidene camphor, 2- Ethylhexyl salicylate/Octisalate), 2-Ethylhexyl 4-

(dimethylamino)benzoate/Padimate O, 2-Hydroxy-4- methoxybenzophenone- 5 -sulfonic acid (Benzophenone-5) and its sodium salt/Sulisobenzone, 2-Hydroxy-4-methoxybenzophenone-5-sulfonic acid (Benzophenone-5) and its sodium salt/Sulisobenzone, 2,2 '-Methylene bis(6- (2H-benzotriazol-2-yl)-4-( 1 , 1 ,3 ,3-tetramethylbutyl)phenol)/Bisoctrizole, Sodium salt of 2,2'-bis(l,4-phenylene)-lH-benzimidazole-4,6-disulfonic acid/Bisdisulizole disodium, 2,2'-(6-(4-Methoxyphenyl)-l,3,5-triazine-2,4- diyl)bis(5-((2-ethylhexyl)oxy)phenol)/Bemotrizinol, Titanium dioxide, Benzoic acid, 2-[4-(diethylamino)-2-hydroxybenzoyl]-, hexylester, 1,3,5- Triazine, 2,4,6-tris(l,l '-biphenyl)-4-yl-, including as nanomaterial and other sunscreens approved for cosmetic use by the legislation of the country where the cosmetic composition will be marketed.

12. A method according to any one of the preceding claims, characterized in that the substances administered for cosmetic use are prepared in a solution selected from a group comprising a cream, an ointment, a water in oil emulsion, an oil in water emulsion or any other type of emulsion, a gel, a paste, a cataplasm, a medicated poultice, a medicated foam or a medicated plaster.

13. A method according to any one of the preceding claims, characterized in that the substances administered for cosmetic use comprise at least one excipient selected from a group comprising petrolatum, fats, silicones, isopropyl myristate, lanolin, lanolin alcohols, lano-vaseline, ethylene oxide derivatives, polyethylene glycols, water, hydrocolloids, starch, methylcellulose, carboxymethylcellulose, gelatin, carbopol, or any other excipient for cosmetic use.

14. A method according to any one of the preceding claims, characterized in that the substances administered for cosmetic use comprise surfactants, preservatives, antioxidants, dyestuffs or fragrances, chosen according to the content of the national regulations of the countries in which will be marketed.