Method for predicting response to a cosmetic treatment comprising retinol

Abstract

Method for predicting response to a cosmetic treatment comprising retinol The present invention relates to a method for predicting the response of a subject's skin to a cosmetic treatment comprising retinol and / or a retinol derivative. It also relates to a method for preventing and / or treating skin signs of aging in a subject. It also relates to a kit and its use for predicting the response of a subject's skin to a cosmetic treatment comprising retinol and / or a retinol derivative.

Description

[0001] DESCRIPTION

[0002] TITLE: Method for predicting response to a cosmetic treatment comprising retinol

[0003] The present invention relates to a method for predicting the response of a subject's skin to a cosmetic treatment comprising retinol and / or a retinol derivative. It also relates to a method for preventing and / or treating skin signs of aging in a subject. It also relates to a kit and its use for predicting the response of a subject's skin to a cosmetic treatment comprising retinol and / or a retinol derivative.

[0004] Vitamin A is known as an anti-wrinkle agent modifying the appearance of the skin and having an anti-aging effect. Retinoids are a class of compounds derived from vitamin A, of which retinol is the best-known cosmetic and biological active ingredient. Retinol does not exert a direct biological effect on tissues such as skin but becomes active after conversion into retinoic acid.

[0005] A recent meta-analysis of 9 vehicle-controlled, double-blind, randomized clinical trials of over-the-counter cosmetic products based on vitamin A demonstrated their efficacy (in particular a significant improvement) on fine lines in only 5 of the 9 studies (Spierings NMK (2021) The Journal of Clinical and Aesthetic Dermatology, 14(9), 33-40). This suggests that certain intrinsic skin parameters may influence the result of applying a retinol-based product on the skin.

[0006] There is therefore a need to find biomarkers to predict the response to a cosmetic treatment comprising retinol. These biomarkers may thus help guide customers identified as responders toward the application of a formula containing retinol in their routine in order to observe an improvement in their wrinkles, i.e. a reduction in their number and / or an attenuation of their appearance.

[0007] This invention responds to this need.

[0008] Surprisingly, the Applicant has now discovered that individuals responding to a cosmetic treatment comprising retinol (also referred to here as "responders"), in particular by an improvement in their wrinkles and / or fine lines, such as crow's feet, under-eye wrinkles,exhibit an overexpression of three proteins (ALDH3A2, NQO2, and RARRES1) involved in the retinol metabolism.

[0009] The overexpression of these proteins prior to any retinol application in these individuals suggests that their skin has a more active retinol transport and metabolism. Their skin would thus be predisposed to respond to retinol thanks to a high expression level of an enzymatic machinery and specific transporters associated with retinol.

[0010] The present invention therefore relates to a method for predicting the response of a subject's skin to a cosmetic treatment comprising retinol and / or a retinol derivative, said method comprising:

[0011] a) measuring the expression level of at least one gene involved in the retinol metabolism chosen from ALDH3A2, NQO2, and RARRES1, in a skin sample from said subject.

[0012] It also relates to a method for preventing and / or treating skin signs of aging in a subject, said method comprising the following steps:

[0013] a) measuring the expression level of at least one gene involved in the retinol metabolism chosen from ALDH3A2, NQO2, and RARRES1, in a skin sample from said subject;

[0014] b) inferring from step a) whether said subject's skin is likely to respond to a cosmetic treatment comprising retinol and / or a retinol derivative when the measured gene(s) is(are) overexpressed, particularly significantly overexpressed, relative to a control; and

[0015] c) if the skin is identified as likely to respond to a treatment comprising retinol and / or a retinol derivative in step b), treating said skin with a cosmetic formulation comprising at least retinol and / or a retinol derivative.

[0016] The present invention also relates to a kit comprising:

[0017] a) means for measuring, in a skin sample, the expression level of at least one gene involved in the retinol metabolism chosen from ALDH3A2, NQO2, and RARRES1; and b) an instruction leaflet.

[0018] It also relates to the use of a kit according to the invention for predicting the response of a subject's skin to a cosmetic treatment comprising retinol and / or a retinol derivative.

[0019] Detailed description of the inventionMethods

[0020] The present invention therefore relates to a method for predicting the response of a subject's skin to a cosmetic treatment comprising retinol and / or a retinol derivative, said method comprising:

[0021] a) measuring the expression level of at least one gene involved in the retinol metabolism chosen from ALDH3A2, NQO2, and RARRES1, in a skin sample from said subject.

[0022] Preferably, the method according to the invention further comprises the following step: b) inferring whether said subject's skin is likely to respond to the cosmetic treatment comprising retinol and / or a retinol derivative when the measured gene(s) is(are) overexpressed, particularly significantly overexpressed, relative to a control.

[0023] The prediction method of the invention is preferably implemented ex vivo.

[0024] According to one preferred embodiment, the cosmetic treatment comprising retinol and / or a retinol derivative comprises the application of a cosmetic composition comprising from 0.05% to 3%, preferably from 0.05% to 2%, preferably from 0.1% to 1% of retinol and / or retinol derivative by weight relative to the total weight of the composition.

[0025] The present invention also relates to a method for preventing and / or treating skin signs of aging in a subject, said method comprising the following steps:

[0026] a) measuring the expression level of at least one gene involved in the retinol metabolism chosen from ALDH3A2, NQO2, and RARRES1, in a skin sample from said subject; b) inferring from step a) whether said subject's skin is likely to respond to a cosmetic treatment comprising retinol and / or a retinol derivative when the measured gene(s) is(are) overexpressed, particularly significantly overexpressed, relative to a control; and c) if the skin is identified as likely to respond to a treatment comprising retinol and / or a retinol derivative in step b), treating said skin with a cosmetic formulation comprising at least retinol and / or a retinol derivative.

[0027] Preferably, the prevention and / or treatment method according to the invention is a cosmetic, in particular non-therapeutic, method.According to one preferred embodiment, the cosmetic composition comprises from 0.05% to 3%, preferably from 0.05% to 2%, preferably from 0.1% to 1% of retinol and / or retinol derivative by weight relative to the total weight of the composition.

[0028] Retinol "derivative" means a natural or synthetic derivative. In particular, such a natural derivative is a compound chosen from retinol isomers, retinoic acid, vitamin A, 9-cis-retinoic acid, retinal, retinyl palmitate, retinyl acetate, and retinyl linoleate. Such a synthetic derivative is a compound chosen from 13-cis-retinoic acid, acitretin, tazarotene, adapalene, and bexarotene.

[0029] The genes according to the invention are typically involved in the metabolism and / or transport of retinol and / or one of its derivatives.

[0030] ALDH3A2 is a gene encoding an enzyme of the aldehyde dehydrogenase (ADH) family having a crucial role in the aldehyde metabolism in different tissues, including skin. This enzyme is necessary for the conversion of retinol into retinoic acid. The human gene has the accession number ID 224 in the NCBI bank (as updated on April 7, 2024).

[0031] NQO2 is a gene encoding the NAD(P)H quinone oxydoreductase enzyme involved in quinone reduction. Some studies suggest that NQO2 is involved in the oxidation of retinol to retinaldehyde and then to retinoic acid. The human gene has the accession number ID 4835 in the NCBI bank (as updated on April 11, 2024).

[0032] RARRES1, also called retinoic acid receptor responder element 1, is a transcription activator through its interaction with retinoic acid receptors. The human gene has the accession number ID 5918 in the NCBI bank (as updated on March 5, 2024).

[0033] The subject is preferably a human subject, preferably a woman with wrinkles and / or fine lines, such as crow's feet, under-eye wrinkles, and aged at least 30 years, preferably at least 40 years, more preferably at least 50 years. Preferably, the subject is a woman aged 30 to 80 years, preferably 40 to 70 years, preferably 50 to 60 years.

[0034] The skin signs of aging are preferably the presence of wrinkles, in particular wrinkles and / or fine lines such as crow's feet, under-eye wrinkles.According to a preferred embodiment of the methods according to the invention, the response to the cosmetic treatment comprising retinol and / or a retinol derivative is the improvement in wrinkles and / or fine lines such as crow's feet, under-eye wrinkles.

[0035] "Improvement in wrinkles and / or fine lines" means a reduction in the number of wrinkles, and / or fine lines, and / or the attenuation of the appearance of wrinkles, and / or fine lines respectively.

[0036] Preferably, the responder exhibits an improvement in their wrinkles and / or fine lines following a treatment comprising retinol and / or a retinol derivative after 4 weeks, after 12 weeks, preferably after 24 weeks of treatment. "Responder" means a subject responding to a cosmetic treatment comprising retinol or a retinol derivative, particularly by the improvement in wrinkles and / or fine lines, such as crow's feet, under-eye wrinkles.

[0037] According to one preferred embodiment of the methods according to the invention, the skin sample from said subject is a sample of the stratum corneum previously obtained by a non-invasive method, in particular by rubbing the surface of the skin or using an adhesive surface on the skin, in particular by D-squame® or Corneofix®.

[0038] “Gene expression level” means the quantity of mRNA produced and / or the quantity of proteins produced by gene expression. This quantity may be expressed as it is, or in concentration or ratio form.

[0039] The expression level of the gene in question can be measured by any suitable technique known from the prior art. To measure the quantity of mRNA, the RNA can be extracted by any RNA extraction method from the skin, then can be processed for a specific mRNA quantification by any mRNA quantification method, for example by quantitative PCR. To measure the quantity of protein, the proteins can be extracted using a buffer suitable for protein extraction from the skin, for example RIPA Buffer or urea buffer, then be processed by any specific protein evaluation method, for example Western blot or ELISA.

[0040] "Overexpressed relative to a control" means that the expression of a gene is significantly greater than a control.

[0041] Preferably, the gene is considered overexpressed when its expression level of said gene is at least 1.2 times, at least 1.3 times, at least 1.4 times, at least 1.5 times, at least 1.6 times,at least 1.7 times, at least 1.8 times, at least 1.9 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 10 times, or at least 13 times greater, preferably significantly greater, than a control.

[0042] According to one preferred embodiment, the expression level of the ALDH3A2 gene is at least 1.2 times, at least 1.3 times, at least 1.4 times, at least 1.5 times greater, preferably significantly greater, than a control.

[0043] According to one preferred embodiment, the expression level of the NQO2 gene is at least 2 times greater, at least 5 times greater, at least 10 times greater, at least 13 times greater, preferably significantly greater, than a control.

[0044] According to a preferred embodiment, the expression level of the RARRES1 gene is at least 1.5 times greater, at least 3 times greater, at least 4 times greater, preferably significantly greater, than a control.

[0045] The control according to the invention is a reference value determined by the mean of the expression level of said gene(s) measured in a defined non-responder population before application of a cosmetic treatment comprising retinol and / or a retinol derivative. Preferably, the defined population is a female population having wrinkles and / or fine lines, such as crow's feet, under-eye wrinkles.

[0046] "Non-responder" means an individual not responding to a cosmetic treatment using a composition, particularly applied topically, comprising retinol and / or a retinol derivative, in particular a subject exhibiting no improvement in wrinkles and / or fine lines, such as crow's feet, under-eye wrinkles.

[0047] According to one preferred embodiment, the subject's skin is treated with a cosmetic composition comprising at least retinol and / or a retinol derivative for at least 4 weeks, at least 12 weeks, preferably at least 24 weeks.

[0048] Preferably, the cosmetic formulation comprising at least retinol and / or a retinol derivative is an anti-aging formulation. It can be applied via any appropriate route of administration, in particular topically. It may, for example, be in cream, lotion, gel, or suspension form.Preferably, the cosmetic composition according to the invention further comprises a physiologically acceptable excipient, i.e. an excipient compatible with the skin and / or mucous membranes.

[0049] The compositions according to the invention may be in any galenic forms conventionally used for a topical application and particularly in the form of anhydrous composition, aqueous, hydroalcoholic solutions, oil-in-water (O / W) or water-in-oil (W / O) or multiple (triple: W / O / W or O / W / O) emulsions, of aqueous gels, or of dispersions of a fatty phase in an aqueous phase using spherules, these spherules may consist of ionic- and / or nonionic-type lipid vesicles (liposomes, niosomes, oleosomes). These compositions are prepared using routine methods.

[0050] The compositions according to the invention may also be in anhydrous form, such as for example in the form of an oil. "Anhydrous composition" denotes a composition containing less than 1% by weight of water, or less than 0.5% of water, and particularly free from water, water not being added during the preparation of the composition but corresponding to the residual water provided by the mixed ingredients.

[0051] When the composition used according to the invention comprises an oily phase, this oily phase preferably contains at least one oil, preferably an oil of plant origin. Oils of plant origin may particularly be liquid fatty acid triglycerides having from 4 to 10 carbon atoms such as heptanoic or octanoic acid triglycerides or, for example, sunflower, corn, soybean, pumpkin, grape seed, sesame, hazelnut, apricot, macadamia, arara, sunflower, castor, avocado oils, caprylic / capric acid triglycerides, jojoba oil, shea butter oil. Preferably, the oil of plant origin is a soybean oil.

[0052] The composition may further comprise at least one adjuvant, particularly those commonly used in the cosmetic field, and particularly chosen from oils; waxes, pigments, fillers, colorants, surfactants, emulsifiers; cosmetic active ingredients, UV filters, polymers, thickeners, film-forming polymers, preservatives, perfumes, bactericides, odor absorbers, antioxidants. The quantities of these various adjuvants are those conventionally used in the field in question, for example from 0.01 to 20% of the total weight of the composition.

[0053] Kit and its use for predicting response to a cosmetic treatment comprising retinol and / or a retinol derivativeThe present invention also relates to a kit comprising:

[0054] a) means for measuring, in a skin sample, the expression level of at least one gene involved in the retinol metabolism chosen from ALDH3A2, NQO2, and RARRES1; and b) an instruction leaflet.

[0055] "Means for measuring the expression level of a gene" means a specific ligand of the protein encoded by said gene, such as for example an antibody, preferably monoclonal, a Fab fragment, an scFv, or a nanobody, specific for this protein. Alternatively, this means a complementary mRNA nucleotide sequence of said gene and specifically hybridizing with the mRNA of said gene, or a fragment thereof hybridizing specifically with the mRNA of said gene, this sequence or this fragment comprising 5 to 50 nucleotides, preferentially 10 to 20 nucleotides, or a pair of primers or a probe of 10 to 60 nucleotides, preferably 15 to 30 nucleotides comprising said sequence or said fragment.

[0056] Preferably, the means for measuring the expression of said gene is at least one specific ligand of the protein encoded by said gene, in particular a pair of capture and detection antibodies specific for the protein encoded by said gene.

[0057] According to one embodiment, the kit according to the invention further comprises a means for collecting the skin sample, such as a cotton swab or an adhesive surface, in particular a D-squame® or Corneofix® disk.

[0058] According to one embodiment, the kit according to the invention further comprises a control, i.e. a sample comprising a known mRNA or protein concentration

[0059] The present invention also relates to the use of a kit according to the invention for predicting the response of a subject's skin to a cosmetic treatment comprising retinol and / or a retinol derivative.

[0060] [Fig. 1] Figure 1 is a box plot showing the pre-treatment ALDH3A2, NQO2, and RARRES1 protein concentration distribution in retinol responders and non-responders.

[0061] The present invention will be described in more detail by the examples hereinbelow.Example 1: clinical study of response to retinol

[0062] Materials & Methods

[0063] 1) Clinical study

[0064] A clinical study was designed wherein 12 people received an emulsion-type formula containing retinol (0.1% or 0.3% retinol). Adhesive strips (D-squame®) were collected at the start of the study, prior to any application of the product, for proteomic analysis. The clinical evaluation was carried out after 16 weeks of daily application.

[0065] 2) Proteomic analysis

[0066] The proteomic data on the adhesive strips collected from the volunteers' faces enabled the differential analysis of more than 1000 proteins by liquid chromatography-tandem mass spectrometry (LC-MS / MS).

[0067] 3) Mass spectrometry data processing.

[0068] The protein identification was carried out by comparative research with Uniprot Homo sapiens reference proteomic data. Finally, a T-test was carried out to compare the relative quantities, before treatment, of the 1013 proteins identified among the 8 responders and the 4 non-responders in order to find potential predictors of response to retinol.

[0069] Results

[0070] A responder is defined as an individual showing a statistically significant improvement in a clinical sign after 16 weeks of product application, in particular an improvement in fine lines.

[0071] Using this criterion, 8 responders and 4 non-responders were identified.

[0072] The proteomic analysis of skin samples collected prior to retinol treatment made it possible to identify 1013 proteins which were quantified. Of these proteins, those which are known to be involved in the retinol metabolism were selected.

[0073] The expression level of these proteins in responders was compared to that in non-responders (Table 1). Table 2 lists the names and Uniprot access numbers of the genes encoding the proteins listed in Table 1 and for which the expression levels were measured[Table 1]

[0074]

[0075] [Table 2]

[0076]

[0077]

[0078] Surprisingly, only three proteins exhibit significant variation (p-value less than 0.05): ALDH3A2, NQO2, and RARRES1 (Fig.1 and indicated in bold in Table 1).

[0079] Thus, the ALDH3A2 expression level is approximately 1.6 times greater in responders than in non-responders. The NQO2 expression level is approximately 13 times greater in responders than in non-responders. Finally, the RARRES1 expression level is approximately 4.2 times greater in responders than in non-responders.

[0080] Conclusion

[0081] Surprisingly, only three proteins (ALDH3A2, NQO2, and RARRES1) are significantly overexpressed in responders among all those involved in the retinol metabolism which were detected.

[0082] These proteins thus constitute biomarkers for which the base concentration in the stratum corneum correlates with the response to retinol.These biomarkers being up-regulated in responders prior to any retinol treatment, it would appear that responders' skin is predisposed to respond more effectively to retinol application by means of an increased ability to transport and metabolize retinol.

[0083] Measuring the expression of these biomarkers in subjects would thus make it possible to guide them or not toward a treatment comprising retinol and / or a retinol derivative based on whether they are identified as responders or non-responders.

Claims

CLAIMS1. A method for predicting the response of a subject's skin to a cosmetic treatment comprising retinol and / or a retinol derivative, said method comprising:a) measuring the expression level of at least one gene involved in the retinol metabolism chosen from ALDH3A2, NQO2, and RARRES1, in a skin sample from said subject.

2. The method according to claim 1, further comprising the following step: b) inferring whether said subject's skin is likely to respond to the cosmetic treatment comprising retinol and / or a retinol derivative when the measured gene(s) is(are) overexpressed, particularly significantly overexpressed, relative to a control.

3. A method for preventing and / or treating skin signs of aging in a subject, said method comprising the following steps:a) measuring the expression level of at least one gene involved in the retinol metabolism chosen from ALDH3A2, NQO2, and RARRES1, in a skin sample from said subject;b) inferring from step a) whether said subject's skin is likely to respond to a cosmetic treatment comprising retinol and / or a retinol derivative when the measured gene(s) is(are) overexpressed, particularly significantly overexpressed, relative to a control; andc) if the skin is identified as likely to respond to a treatment comprising retinol and / or a retinol derivative in step b), treating said skin with a cosmetic formulation comprising at least retinol and / or a retinol derivative.

4. The method according to one of the preceding claims, wherein the response to the cosmetic treatment comprising retinol and / or a retinol derivative is the improvement in wrinkles and / or fine lines, such as crow's feet, under-eye wrinkles.

5. The method according to any one of claims 2 to 4, wherein the control is a reference value determined by the mean of the expression level of said gene(s) measured in a defined non-responder population, in particular a female population exhibiting wrinkles and / or fine lines, such as crow's feet, under-eye wrinkles.

6. The method according to one of the preceding claims, wherein the skin sample from said subject is a sample of the stratum corneum previously obtained by a non-invasive method, in particular by rubbing the skin surface or using an adhesive surface on the skin.

7. The method according to one of the preceding claims, wherein the gene expression level is the quantity of mRNA produced and / or the quantity of proteins produced by the expression of said gene.

8. A kit comprising:a) means for measuring, in a skin sample, the expression level of at least one gene involved in the retinol metabolism chosen from ALDH3A2, NQO2, and RARRES1; and b) an instruction leaflet.

9. The kit according to claim 8, wherein the means for measuring the expression level of at least one gene involved in the retinol metabolism chosen from ALDH3A2, NQO2, and RARRES1 is at least one specific ligand for the protein encoded by said at least one gene, preferably a pair of capture and detection antibodies specific for the protein expressed by said at least one gene.

10. A use of a kit according to claims 8 or 9 for predicting the response of a subject's skin to a cosmetic treatment comprising retinol and / or a retinol derivative.

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