Cosmetic composition containing 13-hydroxy-9(z)-octadecenoic acid for stimulating hair growth

Abstract

The invention relates to a cosmetic composition comprising a modified vegetable oil containing 13-hydroxy-9(Z)-octadecenoic acid. The cosmetic composition is used for stimulating hair growth and for repairing hair fibres.

Description

DESCRIPTION COSMETIC COMPOSITION CONTAINING ACID 13-HYDROXY-9(Z)-OCTADECENOIC FOR HAIR GROWTH STIMULATION

[0001] The present invention relates to a cosmetic composition containing 13-hydroxy-9(Z)-octadecenoic acid (hereinafter abbreviated 13-HOD) for stimulating hair growth and repairing the hair fiber.

[0002] In the context of the present invention, "stimulation of hair growth" means stimulation of hair growth as well as that of beard, eyelashes or eyebrows.

[0003] The hair cycle consists of 3 phases: the anagen phase (growth phase), the catagen phase (regression phase) and the telogen phase (resting phase).

[0004] The anagen phase corresponds to the continuous proliferation of matrix cells in the hair bulb. Cellular activity is intense to ensure keratinization and thus hair growth.

[0005] The catagen phase corresponds to the cessation of mitotic activity in the hair bulb. The hair bulb regresses through apoptosis and detaches from the dermal papilla. Without the dermal papilla, the hair is no longer supplied with blood and grows back into the epidermis.

[0006] The telogen phase is characterized by the shedding of the hair.

[0007] The entry into a new growth cycle depends on signals from the dermal papilla that activate the bulge stem cells and allow the hair follicle to regenerate. Because hair follicles are sensitive to many factors, the duration of the anagen phase can be shortened (and the duration of the telogen phase lengthened) by exposure to external stressors, hormonal changes, or nutritional deficiencies.

[0008] The hair growth cycle is regulated by the interaction between dermal papilla cells and epithelial cells of the hair follicle. Dermal papilla cells can promote and maintain the growth and development of hair follicles by secreting various cytokines and growth factors. These cells play a role important in the regeneration of the hair follicle and its growth by maintaining the hair follicle in the anagen phase.

[0009] During the anagen phase, the hair follicle requires sufficient nutrients to allow cell division of the matrix cells. Vascular endothelial growth factor (VEGF) plays a crucial role in inducing angiogenesis in endothelial cells, the process of new blood vessel growth. Specifically, VEGF induces the proliferation of a capillary network during the anagen phase in the dermal papilla. Angiogenesis leads to improved blood flow to the hair follicle and, consequently, a greater supply of nutrients, thus promoting hair growth.

[0010] Furthermore, VEGF, through binding to the vascular endothelial growth factor receptor 2 (VEGFR-2), activates the extracellular signal-regulatory kinase (ERK) pathway. This pathway, following a series of intracellular phosphorylations, allows the transcription of genes responsible for the proliferation of dermal papilla cells.

[0011] Thus, the cells of the dermal papilla produce VEGF, which will stimulate their proliferation and migration.

[0012] Therefore, VEGF regulates hair growth through its angiogenic effect but also through the VEGFR-2 receptor expressed on dermal papilla cells.

[0013] Furthermore, prostaglandins are lipid-soluble molecules secreted into the extracellular environment that play an important role as paracrine and autocrine signaling agents.

[0014] Prostaglandin E2 (hereafter abbreviated as "PGE2") is linked to hair growth. It is produced from prostaglandin H2 by prostaglandin E synthase. PGE2 is associated with the induction of VEGF. It thus plays a role in the induction of hair growth.

[0015] Prostaglandin D2 (hereafter abbreviated as "PGD2") is a negative regulator of hair growth. PGD2 is produced through the enzymatic conversion of its precursor, prostaglandin H2, by prostaglandin D synthase. PGD2 is an important regulator of inflammation. One of the receptors for PGD2, the GRP44 receptor, is expressed in the outer sheath of the hair follicle. Studies have shown that high levels of PGD2 in human and animal models are associated with inhibition of hair follicle growth, and topical application of PGD2 inhibits hair growth. PGD2 halts hair growth by shortening the anagen phase through activation of the GRP44 receptor, leading to an increased proportion of follicles in the telogen phase and hair follicle miniaturization.

[0016] Hair growth is optimal when there is an increase in PGE2 and a decrease in PGD2 within the hair follicle.

[0017] It is known that the hair growth cycle can be disrupted by various factors, including stress, genetics, vitamin deficiencies, hormones, fatigue, medications, and insufficient blood supply to the scalp. These factors lead to changes in the duration of each phase of hair growth, notably a shortening of the anagen phase. Continuous exposure to pollution and seasonal changes can also cause the hair to enter the telogen phase prematurely.

[0018] Slow hair growth can also result from poor maintenance with inappropriate care and frequent use of heat styling. Finally, eyelashes can be prone to premature shedding during vigorous makeup removal or the use of waterproof mascara.

[0019] Furthermore, aging leads to a shortening of the anagen phase, hair weakening, and a decline in hair quality through the cessation of mitotic activity in keratinocytes and the death of stem cells. Hair loss occurs when cell signaling pathways in hair follicle cells are disrupted, resulting in the induction of apoptosis, changes in the hair cycle, and hair thinning.

[0020] A vegetable oil generally comprises, in mass percentages expressed in relation to the mass of said vegetable oil, between about 95% and about 99% triglycerides, the other constituents of the vegetable oil being minor.

[0021] Triglycerides are glycerides in which the three hydroxyl groups of glycerol are esterified by fatty acids, which may be identical or different. Triglycerides are thus esters derived from one molecule of glycerol and three fatty acid molecules, or in other words, glycerol-fatty acid triesters.

[0022] Depending on the plant species from which it is derived and its extraction method, a vegetable oil is more or less rich in saturated, polyunsaturated, and monounsaturated fatty acids. The distribution of these different fatty acids gives the vegetable oil its specific physical properties (for example, its texture and fluidity).

[0023] Thus, a vegetable oil comprises a mixture of fatty acids, predominantly in the form of triglycerides. Depending on the extraction method, free fatty acids may be present, but in very small quantities. A vegetable oil can be obtained by mechanical extraction, particularly by pressing. It may be refined or unrefined.

[0024] Castor oil is derived from the seeds of the castor bean plant. Ricinoleic acid (also known as 12-hydroxy-9-cis-octadecenoic acid) constitutes approximately 90% of its fatty acids, which are in the form of triglycerides. It has the following chemical structure (1):

[0025] Castor oil, as well as ricinoleic acid, are known to promote hair growth.

[0026] Furthermore, patent applications WO 2021 / 209437 Al and WO 2021 / 209442 Al describe natural cosmetic compositions for stimulating hair growth that are free of any preservatives. These compositions include, among other things active ingredients, ricinoleic acid in combination with arachidonate and possibly diosgenin.

[0027] However, given consumer needs in the field of cosmetics, there is still a real interest in developing new cosmetic compositions that stimulate hair growth and / or repair the hair fiber, with very effective results in order to effectively address the hair cycle disorders mentioned above.

[0028] The inventors made a truly surprising discovery: a cosmetic composition containing 13-HOD as its active ingredient fully met all these objectives. As will be detailed in the experimental section below, the inventors demonstrated all the beneficial effects of 13-HOD on stimulating hair growth and repairing the hair fiber.

[0029] 13-HOD has the following chemical structure (2):

[0030] 13-HOD is a hydroxylated derivative of linoleic acid, which is a fatty acid. Therefore, 13-HOD is a hydroxylated fatty acid.

[0031] Linoleic acid has the following chemical structure (3):

[0032] Linoleic acid is present, in varying quantities, in many vegetable oils (for example grapeseed or sunflower oil), mainly in the form of triglycerides and therefore very little in the form of free fatty acids.

[0033] Unlike linoleic acid, vegetable oils do not naturally contain 13-HOD. In other words, there is no naturally occurring 13-HOD.

[0034] 13-HOD is known for its use as an intermediate in the preparation of lactones, which are chemical compounds used in various fields. very diverse applications such as food, polymers, agriculture and especially in cosmetics and perfumery.

[0035] This is why the synthetic routes of 13-HOD are perfectly known and mastered by those skilled in the art, given that they are notably implemented during lactone preparation processes.

[0036] For example, 13-HOD can be obtained from pure linoleic acid. In this regard, US patent 11,499,171 B2 describes a biocatalytic synthesis route for 13-HOD from pure linoleic acid in the presence of a modified microorganism producing a protein with a defined amino acid sequence and having 13-linoleate hydratase activity.

[0037] As explained above, vegetable oils do not naturally contain 13-HOD. However, processing a natural vegetable oil can yield a modified vegetable oil containing 13-HOD.

[0038] In this regard, patent application IT 2019 / 00015713 Al describes the following steps in the transformation of a vegetable oil that result in a modified vegetable oil containing 13-HOD: - the biocatalytic hydrolysis of triglycerides present in vegetable oil in order to obtain fatty acids (in particular oleic, linoleic and linolenic acid) in free form, by means of an enzyme (lipase); - hydration by bioconversion of the double bonds of the fatty acids thus released in the previous step by means of bacterial fermentation with the use of probiotic microorganisms expressing hydratase enzymes in order to transform linoleic acid into 13-HOD.

[0039] The invention thus relates to a cosmetic composition characterized in that it comprises at least one modified vegetable oil containing 13-HOD.

[0040] For the purposes of this invention, "modified vegetable oil" means a vegetable oil that does not exist in a natural form. Indeed, as explained above, vegetable oils do not naturally contain 13-HOD.

[0041] Modified vegetable oil may comprise, in mass percentages expressed relative to the total mass of free fatty acids contained in said modified vegetable oil, at least 10%, preferably at least 20% and even more preferably at least 40%, of 13-HOD. The modified vegetable oil may comprise, in mass percentages expressed in relation to the total mass of free fatty acids contained in said modified vegetable oil, between 10% and 80%, preferably between 20% and 70%, more preferably between 30% and 60%, of 13-HOD.

[0042] Indeed, unlike so-called "natural" vegetable oils, which, as explained above, are characterized by almost all of their fatty acids being in triglyceride form, modified vegetable oil can be a vegetable oil specifically formulated to contain fatty acids (including 13-HOD) primarily in free form. For example, modified vegetable oil can contain between 80% and 100%, preferably between 90% and 99%, of free fatty acids, relative to its total fatty acid mass.

[0043] Modified vegetable oil may contain small amounts of triglycerides. For example, modified vegetable oil may contain, as a mass percentage relative to the total mass of said modified vegetable oil, less than 20%, preferably less than 10%, and more preferably less than 5%, of triglycerides. Modified vegetable oil may also contain, as a mass percentage relative to the total mass of said modified vegetable oil, between 0.5% and 20%, preferably between 1% and 15%, and more preferably between 3% and 10%, of triglycerides.

[0044] Preferably, the modified vegetable oil may contain no glycerol or only traces of glycerol. The mass percentage of glycerol, expressed as a percentage of the total mass of said vegetable oil, may be at most 5%, preferably 2%. In one embodiment of the invention, said mass percentage of glycerol may be between 0.05% and 5%, preferably between 0.1% and 3%, more preferably between 0.2% and 2%.

[0045] Vegetable oil can be chosen from the group consisting of grapeseed, safflower, borage, peanut, wheat germ, corn, sunflower, blackberry, black cumin, evening primrose, pumpkin seed, hemp, cottonseed, walnut, soybean, argan, and rapeseed oils, used alone or in blends. These vegetable oils naturally have high linoleic acid content. Their processing, using methods perfectly within the capabilities of those skilled in the art, allows to obtain modified vegetable oils containing 13-HOD. Preferably, the vegetable oil is grapeseed oil.

[0046] The cosmetic composition according to the invention may comprise, in mass percentages expressed in relation to the total mass of said cosmetic composition, between 0.0001% and 1%, preferably between 0.001% and 0.1%, of 13-HOD.

[0047] When the cosmetic composition includes a modified vegetable oil containing 13-HOD, the mass percentage of said modified vegetable oil, expressed in relation to the total mass of said cosmetic composition, may be between 0.0005% and 5%, preferably between 0.005% and 0.5%.

[0048] The cosmetic composition according to the invention may further comprise at least one adjuvant chosen from among those commonly used in the field of cosmetics. These may include, for example, emulsifiers, surfactants, hydrophilic or lipophilic gelling agents, preservatives, antioxidants, solvents, perfumes, fillers, pigments or coloring matter, ultraviolet filters, or pH stabilizers.

[0049] The mass percentages of these various adjuvants are those typically used in the cosmetics industry. For example, these mass percentages range from 0.01% to 20% of the total mass of the cosmetic composition.

[0050] These adjuvants, depending on their nature, can be introduced into the oily phase, the aqueous phase or the lipid vesicles of the cosmetic composition according to the invention.

[0051] In any case, these adjuvants, as well as their quantities, are chosen so as not to impair the desired properties of 13-HOD.

[0052] Examples of hydrophilic gelling agents include carboxyvinyl polymers, acrylic copolymers such as acrylate / alkylacrylate copolymers, polysaccharides, natural gums and clays, and examples of lipophilic gelling agents include modified clays such as bentones and hydrophobic silica.

[0053] Examples of fillers include polymethyl methacrylate microspheres, acrylate-acrylate copolymer powders, expanded powders such as hollow microspheres, and in particular, powders of natural organic materials such as starch powders, especially corn, wheat or rice starches, crosslinked or not, such as starch powders crosslinked by octenylsuccinate anhydride, silica, metal oxides such as titanium dioxide or zinc oxide, mica, and mixtures thereof.

[0054] Examples of antioxidants include tocopherol (in other words, the aforementioned vitamin E) and its esters, particularly tocopherol acetate, ascorbic acid (in other words, the aforementioned vitamin C) and its derivatives, particularly ascorbyl magnesium phosphate, ascorbyl glucoside and ascorbyl tetraisopalmitate, ferulic acid, serine, ellagic acid, phloretin and mixtures thereof.

[0055] The cosmetic composition according to the invention may be more or less fluid and have the appearance of a white or colored cream, an ointment, a milk, a lotion, a solution, a shampoo, a serum, a paste, a mask, or even a mousse. The cosmetic composition according to the invention may also be in solid form, for example, as a stick.

[0056] The cosmetic composition according to the invention can be presented in all the galenic forms normally used in the field of cosmetics.

[0057] It can be, for example, in the form of an aqueous or oily solution possibly gelled, a lotion-type dispersion possibly biphasic, an emulsion obtained by dispersing an oily phase in an aqueous phase (O / W) or conversely an aqueous phase in an oily phase (W / O), or even a triple emulsion (W / O / W or W / O / O) or a vesicular dispersion of ionic and / or non-ionic type.

[0058] The cosmetic composition according to the invention can be prepared using conventional methods. For example, when the cosmetic composition according to the invention is an emulsion, the mass percentage of the oil phase can be between 0.5% and 80%, and preferably between 5% and 50%, relative to the total mass of said cosmetic composition.

[0059] The oils, emulsifiers and co-emulsifiers used in the cosmetic composition according to the invention in the form of an emulsion are chosen from those classically used in the field concerned.

[0060] The mass percentage of emulsifier and co-emulsifier may be between 0.1% and 30%, preferably between 0.5% and 20%, relative to the total mass of said cosmetic composition.

[0061] Emulsifiers and co-emulsifiers can be chosen from fatty acid and glycerin esters such as glyceryl stearate, sucrose esters and phospholipids.

[0062] The cosmetic composition according to the invention may further comprise at least one oil selected from vegetable oils (e.g. almond oil, apricot oil, liquid fraction of shea butter, avocado and soybean oil) and synthetic oils (e.g. isononyl isononanoate, pentaerythrityl tetraoctanoate).

[0063] The cosmetic composition according to the invention may further comprise fats selected from fatty alcohols (for example cetyl or stearyl alcohol), fatty acids (for example stearic acid) and waxes (for example carnauba wax, ozokerite or beeswax).

[0064] The invention also relates to a non-therapeutic cosmetic process for stimulating hair growth, characterized in that it includes a step of applying a cosmetic composition comprising 13-HOD as an active ingredient to a specific hairy area.

[0065] The invention also relates to a non-therapeutic cosmetic process for stimulating hair growth, characterized in that it includes a step of applying the cosmetic composition according to the invention, as described above, to a specific hairy area.

[0066] The defined hair-bearing area may be all or part of the scalp, chin, eyebrows, or the free edges of the eyelids where the eyelashes are implanted. In other words, the cosmetic composition comprising 13-HOD as its active ingredient, in particular the cosmetic composition according to the invention, may be applied to all or part of the scalp, chin, eyebrows, or eyelashes.

[0067] Preferably, the hair-bearing area is the scalp.

[0068] In one embodiment of the invention, a cosmetic composition comprising 13-HOD as its active ingredient, in particular the cosmetic composition according to the invention, is used to stimulate eyelash growth. Therefore, the invention also relates to a makeup composition, preferably a mascara composition, characterized in that it comprises a cosmetic composition comprising 13-HOD as its active ingredient, in particular the cosmetic composition according to the invention as described above.

[0069] The invention also relates to a non-therapeutic cosmetic use of 13-HOD to stimulate hair growth.

[0070] The invention also relates to a non-therapeutic cosmetic process for repairing hair fibers, characterized in that it includes a step of applying to all or part of the scalp a cosmetic composition comprising 13-HOD as an active ingredient.

[0071] The invention also relates to a non-therapeutic cosmetic process for repairing hair fibers, characterized in that it includes a step of applying the cosmetic composition according to the invention, as described above, to all or part of the scalp.

[0072] The invention also relates to a non-therapeutic cosmetic use of 13-HOD to repair the hair fiber.

[0073] The invention and its advantages are illustrated in the examples below.

[0074] Examples:

[0075] Presentation of compound A and oils A to C:

[0076] Compound A:

[0077] Compound A contained, in mass percentages expressed relative to the total mass of said compound A: - 71.8% of 13-HOD; - 28.2% linoleic acid.

[0078] This compound A was obtained from pure linoleic acid which was subjected to enzymatic biocatalysis such that 71.8% of the linoleic acid was transformed into 13-HOD.

[0079] In the modified grapeseed oils A through C described below, the fatty acids were essentially in free form. This is because all three of these modified grapeseed oils A through C result from the complete conversion of the triglycerides contained in the grapeseed oil from which they are derived.

[0080] Oil A:

[0081] Oil A was a modified grape seed oil which contained in mass percentages expressed relative to the total mass of free fatty acids of said oil A: - 45.6% of 13-HOD; - 19.5% linoleic acid; - 19.1% oleic acid; - 10.6% palmitic acid; - 5.2% stearic acid.

[0082] Oil B:

[0083] Oil B was a modified grapeseed oil which contained, in mass percentages expressed relative to the total mass of free fatty acids in said oil B: - 46.9% of 13-HOD; - 12.8% linoleic acid; - 22.5% oleic acid; - 11.6% palmitic acid; - 6.2% stearic acid.

[0084] Oil C:

[0085] Oil C was a modified grape seed oil which contained, in mass percentages expressed relative to the total mass of free fatty acids of said oil C: - 53.3% of 13-HOD; - 21.2% linoleic acid; - 15.3% oleic acid; - 6.9% palmitic acid; - 3.3% stearic acid.

[0086] Thus, in modified grape seed oils A to C, 13-HOD was the predominant fatty acid among the free fatty acids contained in said modified grape seed oils A to C.

[0087] L ère Series of experiments: effect of compound A on the secretion of PGE2 and PGD2

[0088] The following IA to IC and Control 1 trials were performed.

[0089] In each of the AI ​​to IC and Control 1 trials, dermal papilla cells from hair follicles were cultured in a commercially available culture medium from Promocell under the trade name "Follicle Dermal Papilla Cell Growth Medium" and placed in a humid atmosphere containing 5% CO2 at 37°C.

[0090] AI Test:

[0091] Compound A was diluted in dimethyl sulfoxide (hereinafter abbreviated as "DMSO") to obtain a stock solution of said compound A having a concentration of said compound A of 10 mg / mL.

[0092] The stock solution of compound A was then appropriately diluted in DMSO such that: - that a dilute solution of said compound A was obtained and then added to the aforementioned culture medium, - the mass percentage of compound A expressed in relation to the mass of said culture medium was 0.0005% (i.e. 0.000359% of 13-HOD taking into account that compound A comprised 71.8% of 13-HOD).

[0093] After 48 hours of treatment of dermal papilla cells with compound A at a mass concentration of 0.0005%, the supernatants were recovered. The amounts of secreted PGE2 and PGD2 were measured by enzyme-linked immunosorbent assays (ELISAs) and normalized by cell viability.

[0094] Test IB:

[0095] Ricinoleic acid was diluted in DMSO to obtain a ricinoleic acid stock solution with a ricinoleic acid concentration of 10 mg / mL.

[0096] The ricinoleic acid stock solution was then appropriately diluted in DMSO such that: - that a dilute solution of ricinoleic acid was obtained and then added to the aforementioned culture medium, - the mass percentage of ricinoleic acid expressed in relation to the mass of said culture medium was 0.001%.

[0097] After 48 hours of treatment of dermal papilla cells with ricinoleic acid at a mass concentration of 0.001%, the supernatants were collected. The amounts of secreted PGE2 and PGD2 were measured by ELISA assays and normalized by cell viability.

[0098] IC Test:

[0099] Ricinoleic acid was diluted in DMSO to obtain a ricinoleic acid stock solution with a ricinoleic acid concentration of 10 mg / mL.

[0100] The ricinoleic acid stock solution was then appropriately diluted in DMSO such that: - that a dilute solution of ricinoleic acid was obtained and then added to said culture medium, - the mass percentage of ricinoleic acid expressed in relation to the mass of said culture medium was 0.002%.

[0101] After 48 hours of treatment of dermal papilla cells with ricinoleic acid at a mass concentration of 0.002%, the supernatants were collected. The amounts of secreted PGE2 and PGD2 were measured by ELISA assays and normalized by cell viability.

[0102] Test Control 1:

[0103] Furthermore, a Control Trial 1 was performed by adding no active ingredient to the culture medium in which the dermal papilla cells were cultured. After 48 hours of culture of said dermal papilla cells, the supernatants were collected. The quantities of secreted PGE2 and PGD2 were measured using ELISA assays and normalized by cell viability.

[0104] Figure 1 is a graph representing the amounts of PGE2 and PGD2 secreted by dermal papilla cells during IA to IC trials, said amounts being expressed as percentages relative to the amounts of PGE2 and PGD2 secreted during Control Trial 1 and normalized to 100%.

[0105] As shown in Figure 1, a very significant amount of PGE2 was secreted during trial IA, unlike in trials IB and IC. This means that, unlike ricinoleic acid, 13-HOD induced a significant increase in PGE2 secretion, even at a much lower concentration than ricinoleic acid (0.000359% versus 0.001% and 0.002%).

[0106] Furthermore, in the IA trial, very little PGD2 was secreted. 13-HOD did not induce significant PGD2 secretion.

[0107] This l ère A series of experiments demonstrates that 13-HOD is more effective than ricinoleic acid in stimulating hair follicle growth. Indeed, treatment with 13-HOD resulted in a highly significant increase in PGE2 secretion and a low secretion of PGD2.

[0108] 2 èmeSeries of experiments: Effect of compound A on VEGF secretion

[0109] The following tests 2A to 2C and Control 2 have been carried out.

[0110] In each of the trials 2A to 2C and Control 2, dermal papilla cells from hair follicles were cultured in a culture medium identical to that of the l ère series of experiments which were placed in a humid atmosphere containing 5% CO2 at 37°C. [YES] Test 2A:

[0112] The preparation for test 2A was identical to that of test IA detailed above.

[0113] After 48 hours of treatment of dermal papilla cells with compound A at a mass concentration of 0.0005%, the supernatants were recovered. The amount of VEGF secreted was measured by assays and normalized by cell viability.

[0114] Test 2B:

[0115] The preparation for test 2B was identical to that of test IB detailed above.

[0116] After 48 hours of treatment of dermal papilla cells with ricinoleic acid at a mass concentration of 0.001%, the supernatants were recovered. The amount of VEGF secreted was measured using ELISA assays and normalized by cell viability.

[0117] Test 2C:

[0118] The preparation for test 2C was identical to that for test IC detailed above.

[0119] After 48 hours of treatment of dermal papilla cells with ricinoleic acid at a mass concentration of 0.002%, the supernatants were collected. The amount of VEGF secreted was measured using ELISA assays and normalized by cell viability.

[0120] Test Control 2:

[0121] In addition, a Control Trial 2 was carried out by adding no active ingredient to the culture medium in which the dermal papilla cells were grown.

[0122] After 48 hours of culture of the dermal papilla cells, the supernatants were collected. The amount of VEGF secreted was measured using ELISA assays and normalized by cell viability.

[0123] Figure 2 is a graph representing the amount of VEGF secreted into dermal papilla cells during trials 2A to 2C, with the amount of VEGF secreted expressed as a percentage relative to the amount of VEGF secreted during Control Trial 2 and normalized to 100%.

[0124] As shown in Figure 2, in trial 2A, 13-HOD significantly stimulated VEGF secretion. This significant stimulation of VEGF secretion was also observed in trial 2C, with ricinoleic acid, but at a much higher concentration (0.002% versus 0.000359%).

[0125] 13-HOD thus has an effect on the secretion of VEGF which, as explained above, has a beneficial effect on hair growth.

[0126] 3 ème Series of experiments: effect of oil A on PGE2 secretion

[0127] The following tests 3A to 3C and Control 3 have been carried out.

[0128] In each of the trials 3A to 3C and Control 3, dermal papilla cells from hair follicles were cultured in a culture medium identical to that of the l ère series of experiments which were placed in a humid atmosphere containing 5% CO2 at 37°C.

[0129] Test 3A:

[0130] Oil A was diluted in DMSO to obtain a stock solution of said oil A with a concentration of said oil A of 10 mg / mL.

[0131] The stock solution of oil A was appropriately diluted in DMSO such that: - that a diluted solution of said oil A was obtained and then added to the aforementioned culture medium, - the mass percentage of oil A expressed in relation to the mass of said culture medium was 0.001% (i.e. 0.000456% of 13-HOD, if we consider the very small mass of unsaponifiables present in said oil A as negligible and taking into account that oil A contained essentially free fatty acids, of which 13-HOD had a mass content of 45.6%).

[0132] After 48 hours of treatment of dermal papilla cells with oil A at a mass concentration of 0.001%, the supernatants were collected. The amounts of secreted PGE2 were measured by ELISA assays and normalized by cell viability.

[0133] Test 3B:

[0134] Oil A was diluted in DMSO to obtain a stock solution of said oil A with a concentration of said oil A of 10 mg / mL.

[0135] The stock solution of oil A was appropriately diluted in DMSO such that: - that a diluted solution of said oil A was obtained and then added to the aforementioned culture medium, - the mass percentage of oil A expressed relative to the mass of said medium culture was 0.002% (i.e. 0.000912% of 13-HOD, if we consider the very small mass of unsaponifiables present in said oil A as negligible and taking into account that oil A contained essentially free fatty acids, of which 13-HOD had a mass content of 45.6%).

[0136] After 48 hours of treatment of dermal papilla cells with oil A at a mass concentration of 0.002%, the supernatants were collected. The amounts of PGE2 secreted were measured by ELISA assays and normalized by cell viability.

[0137] Test 3C:

[0138] Castor oil was diluted in DMSO to obtain a stock solution of said castor oil with a concentration of said castor oil of 10 mg / mL.

[0139] The castor oil stock solution was appropriately diluted in DMSO as follows: - that a diluted solution of said castor oil was obtained and then added to the aforementioned culture medium, - the mass percentage of castor oil expressed in relation to the mass of said culture medium was 0.001% (i.e. 0.0009% of ricinoleic acid taking into account that castor oil comprised 90% ricinoleic acid and if we neglect the unsaponifiables present in said castor oil).

[0140] After 48 hours of treatment of dermal papilla cells with castor oil at a mass concentration of 0.001%, the supernatants were collected. The amounts of secreted PGE2 were measured by ELISA assays and normalized by cell viability.

[0141] Test Control 3:

[0142] Furthermore, a Control Trial 3 was performed by adding no active ingredient to the culture medium in which the dermal papilla cells were grown. After 48 hours of culture of these dermal papilla cells, the supernatants were collected. The amounts of PGE2 secreted were measured using ELISA assays.

[0143] Figure 3 is a graph representing the amounts of PGE2 secreted by dermal papilla cells during tests 3A to 3C, said amounts expressed as percentages relative to the amount of PGE2 secreted during Control Test 3 and normalized to 100%.

[0144] As shown in the graph in Figure 3, a significant amount of PGE2 was secreted during trials 3A and 3B, unlike in trial 3C. This means that, unlike castor oil which contains ricinoleic acid, oil A, which contained 13-HOD, induced a significant increase in PGE2 secretion, which has a beneficial effect on hair growth.

[0145] 4 ème Series of experiments: Effect of oil C on VEGF expression:

[0146] Tests 4A to 4C and the following Control 4 have been carried out.

[0147] In each of the trials 4A to 4C and Control 4, dermal papilla cells from hair follicles were cultured in a culture medium identical to that of the 1st series of experiments which was placed in a humid atmosphere containing 5% CO2 at 37°C.

[0148] Test 4A:

[0149] Oil C was diluted in DMSO to obtain a stock solution of said oil C with a concentration of said oil C of 10 mg / mL.

[0150] The stock solution of oil C was appropriately diluted in DMSO such that: - that a diluted solution of said oil C was obtained and then added to the aforementioned culture medium, - the mass percentage of oil C expressed in relation to the mass of said culture medium was 0.001% (i.e. 0.000533% of 13-HOD, if we consider the very small mass of unsaponifiables present in said oil C as negligible and taking into account that oil C contained essentially free fatty acids, of which 13-HOD had a mass content of 53.3%).

[0151] After 24 hours of treatment of dermal papilla cells with oil C at a mass concentration of 0.001%, the cells were washed and collected. Then, RNA extraction was performed using Trizol® and chloroform. After quantification, the RNA was reverse-transcribed into cDNA to assess VEGF gene expression. Ct values ​​were normalized to those of the reference gene GAPDH.

[0152] Test 4B:

[0153] Oil C was diluted in DMSO to obtain a stock solution of said oil C with a concentration of said oil A of 10 mg / mL.

[0154] The stock solution of oil C was appropriately diluted in DMSO such that: - that a diluted solution of said oil C was obtained and then added to the aforementioned culture medium, - the mass percentage of oil C expressed in relation to the mass of said culture medium was 0.002% (i.e. 0.001066% of 13-HOD if we consider the very small mass of unsaponifiables present in said oil C as negligible and taking into account that oil C contained essentially free fatty acids, of which 13-HOD had a mass content of 53.3%; which corresponded to a concentration of 36 pmol / L of 13-HOD).

[0155] After 24 hours of treatment of dermal papilla cells with oil C at a mass concentration of 0.002%, the cells were washed and collected. RNA extraction was then performed using Trizol® and chloroform. After quantification, the RNA was reverse-transcribed to cDNA to assess VEGF gene expression. Ct values ​​were normalized to those of the reference gene GAPDH.

[0156] Test 4C:

[0157] Minoxidil is a vasodilator and antihypertensive medication initially used to treat high blood pressure. One of the side effects of this medication has been observed to be increased hair growth. Therefore, it is known to apply the Minoxidil applied to the scalp to slow hair loss and may even promote regrowth. Therefore, due to its recognized application in stimulating hair growth, minoxidil constitutes a comparable compound to the present invention.

[0158] Minoxidil was appropriately diluted in the aforementioned culture medium such that the concentration of minoxidil in said culture medium was 100 pmol / L.

[0159] After 24 hours of treatment of dermal papilla cells with minoxidil at a concentration of 100 pmol / L, the cells were washed and collected. RNA extraction was then performed using Trizol® and chloroform. After quantification, the RNA was reverse-transcribed into cDNA to assess VEGF gene expression. Ct values ​​were normalized to those of the reference gene GAPDH.

[0160] Test Control 4:

[0161] Furthermore, a Control 4 trial was performed by adding no active ingredient to the culture medium in which the dermal papilla cells were cultured. After 24 hours of culture, the dermal papilla cells were washed and collected. RNA extraction was then performed using Trizol® and chloroform. After quantification, the RNA was reverse-transcribed into cDNA to assess VEGF gene expression. Ct values ​​were normalized to those of the reference gene GAPDH.

[0162] Figure 4 is a graph representing VEGF gene expression in dermal papilla cells during assays 4A to 4C, with VEGF gene expression expressed as a percentage relative to VEGF gene expression obtained during Control Assay 4 and normalized to 100%.

[0163] Based on the graph in Figure 4, it can be seen that in trial 4B, oil C containing 13-HOD significantly stimulated VEGF gene expression (+20%). Minoxidil at a concentration of 100 pmol / L significantly stimulated VEGF expression in dermal papilla cells by 30%. The difference observed in trials 4B and 4C between the effect of minoxidil and the effect of oil C containing 13-HOD was not significant. The effect of these two compounds is therefore similar at a concentration of 13-HOD three times lower (i.e., 36 pmol / L versus 100 pmol / L).

[0164] 5 ème Series of experiments: Effect of oil B on the growth of cultured hair follicles:

[0165] The following 5A tests and Control 5 were carried out.

[0166] For these 5A and Control 5 trials, the culture medium was the SFM of keratinocytes marketed by the company THERMO FISHER SCIENTIFIC and which was placed in a humid atmosphere containing 5% CO2 at 37°C.

[0167] Test 5A:

[0168] Oil B was diluted in DMSO to obtain a stock solution of said oil B with a concentration of said oil B of 10 mg / mL.

[0169] The stock solution of oil B was appropriately diluted in DMSO such that: - that a diluted solution of said oil B was obtained and then added to the culture medium as detailed in previous experiments, - the mass percentage of oil B expressed in relation to the mass of said culture medium was 0.0005% (i.e. 0.0002345% of 13-HOD, if we consider the very small mass of unsaponifiables present in said oil B as negligible and taking into account that oil B contained essentially free fatty acids, of which 13-HOD had a mass content of 46.9%).

[0170] Hair follicles were extracted from surgical residues and cultured in the culture medium which included 0.0005% of oil B.

[0171] At 3 and 7 days after the start of the experiment, half of the culture medium containing 0.0005% oil B was replaced. Hair follicles were cultured for a total of 10 days. Their lengths were measured after 0, 1, 3, 7, and 10 days of the experiment by microscopy image analysis.

[0172] Test 5 Control:

[0173] In addition, a Control Trial 5 was conducted by adding no active ingredient to the culture medium in which the hair follicles were grown. As with Trial 5A, half of the culture medium was replaced at 3 and 7 days after the start of the experiment. The follicles were cultured for a total of 10 Their lengths were measured after 0, 1, 3, 7 and 10 days of experimentation, by microscopy image analysis.

[0174] Figure 5 is a graph of the change in hair follicle length in trial 5A and trial Control 5 over time. More specifically, this length is expressed as a percentage relative to the hair follicle length of trial Control 5 at the beginning of the experiment (i.e., Day 0) and normalized to 100%.

[0175] Based on the shape of the curves in trial 5A and trial Control 5, it can be seen that oil B containing 13-HOD significantly stimulates the growth of hair follicles in culture from 7 days up to 10 days of culture (an increase of 14% and 16% respectively after 7 days and 10 days of culture compared to trial Control 5).

[0176] 6 ème Series of experiments: Effect of oil C on hair growth on skin excipients:

[0177] The following tests 6A, 6B and Control 6 were carried out.

[0178] For these trials 6A, 6B and Control 6, the culture medium was a culture medium suitable for hair growth on skin expiants and was placed in a humid atmosphere containing 5% CO2 at 37°C.

[0179] Tests 6A and 6B:

[0180] During tests 6A and 6B, the following cosmetic composition was used. The list of its constituents and their mass percentages, expressed relative to the total mass of said composition, are detailed in Table 1 below. Table 1

[0181] For test 6A, the active ingredient tested was oil C and for test 6B, the active ingredient tested was castor oil.

[0182] Skin expiants (77-year-old female donor) with at least 3 hair follicles were cultured in the culture medium.

[0183] For trials 6A and 6B, the cosmetic composition detailed above was applied daily for 5 days to these cultured skin excipients. Specifically, 2 mg of said composition were applied per cm² 2 skin explant.

[0184] Hair length was measured after 2 and 5 days of such treatment.

[0185] Test Control 6:

[0186] For control trial 6, skin expiants (77-year-old female donor) with at least 3 hair follicles were cultured in the culture medium.

[0187] No composition was applied to these skin expansives.

[0188] Figure 6 is a graph that represents, for trials 6A, 6B and Control 6, the relative hair growth between days 2 and 5 of culture, expressed as a percentage relative to the hair length on day 2.

[0189] As shown in Figure 6, the relative hair growth measured on the untreated subjects is 102% (i.e., Control 6 trial). This rate is 125% for the subjects in Test 6A, i.e., the subjects treated with oil C containing 13-HOD, and is significantly higher than that of the subjects in Control 6 trial.

[0190] The hair growth rate in trial 6A was higher than in trial 6B, which used castor oil (125% versus 113%). Oil C had a greater beneficial effect on hair growth than castor oil.

[0191] Thus, at equivalent concentrations, oil C containing 13-HOD has a greater stimulatory effect on hair growth than castor oil.

[0192] 7 èmeSeries of experiments: Effect of oil C, used in cosmetic compositions that were rinsed off, on the porosity of the hair cuticle:

[0193] The cuticle is a waterproof, scale-like protective layer that covers the hair shaft.

[0194] Tests 7a to 7D and Control 7 were carried out.

[0195] For each of these trials, 3 strands of hair from a Caucasian donor were used.

[0196] The hair strands in tests 7A to 7D were exposed to heat stress at 95°C for one hour. The strands in Control test 7 were not exposed to such heat stress.

[0197] Tests 7A and 7B:

[0198] During tests 7A and 7B, the following cosmetic compositions 1 and 2 were used respectively. The list of their constituents and their mass percentages, expressed relative to the total mass of the composition in question, are detailed in Table 2 below. Table 2

[0199] For tests 7A and 7B, compositions 1 and 2 were applied to damp hair strands for 10 minutes, respectively. These strands were then washed three times with ultrapure water. The application of compositions 1 and 2 and the three washes were repeated twice more before a final air-drying step.

[0200] The hair strands were then soaked in a fluorophore solution (specifically, a fluorescein solution) and cryopreserved. Cross-sections were prepared. The fluorescent signal was visualized by microscopy and quantified by image analysis.

[0201] Test 7C:

[0202] No composition was applied to the hair strands in test 7C. The hair strands were washed three times with ultrapure water and then air-dried. They were then soaked in a fluorescein solution and cryopreserved. Cross-sections were prepared. The fluorescent signal was visualized by microscopy and quantified by image analysis.

[0203] 7D Test:

[0204] During the 7D trial, a cosmetic composition known as a "placebo" was used. The list of its constituents and their mass percentages, expressed relative to the total mass of the cosmetic composition known as a "placebo," are detailed in Table 3 below. Table 3

[0205] For the 7D trial, the so-called "placebo" composition was applied to damp hair strands for 10 minutes. Then, these hair strands were washed three times with ultrapure water. These steps of applying the "placebo" composition and washing three times were repeated twice more before a final air-drying step.

[0206] The hair strands were then soaked in a fluorescein solution and cryopreserved. Cross-sections were prepared. The fluorescent signal was visualized by microscopy and quantified by image analysis.

[0207] Test Control 7:

[0208] The hair strands in Control 7 were not subjected to heat treatment. Furthermore, no cosmetic composition was applied to them. The hair strands were washed three times with ultrapure water and then air-dried. Next, they were soaked in a fluorescein solution and then cryopreserved. Cross-sections were prepared. The fluorescent signal was visualized by microscopy and quantified by image analysis.

[0209] Figure 7 is a graph which represents, for tests 7A to 7D and Control 7, the diffusion of the fluorophore within the fibers of the hair strands, said diffusion of the fluorophore being the penetration distance of the fluorophore within said fibers expressed as a percentage relative to the diameter of said fibers.

[0210] In the absence of heat stress, the hair strands in Test Control 7 have fibers whose structural integrity prevents fluorophore diffusion. The fluorophore diffusion rate is 10.78%.

[0211] After heat stress, the fibers of the hair strands became porous (in other words, they were no longer intact) and the diffusion of the fluorophore increased by 88%.

[0212] The higher the diffusion of the fluorophore, the more porous the hair strand fibers become.

[0213] That is why for test 7C, the percentage of diffusion of the fluorophore is 20.25%.

[0214] Treating hair strands with a cosmetic composition containing 13-HOD-containing oil C (specifically, tests 7A and 7B) restored the integrity of the hair fibers. Indeed, the fluorophore diffusion percentages were 11.88% and 10.31% for tests 7A and 7B, respectively.

[0215] Treating the hair strands with a so-called "placebo" composition did not restore the integrity of the hair fibers to the same extent. Indeed, the fluorophore diffusion rate in the 7D trial was 14.15%.

[0216] Thus, treating hair strands with a cosmetic composition containing 13-HOD-based oil C restored the integrity of the hair fibers with greater efficacy than the placebo composition. Indeed, fluorophore diffusion decreased by 88% and 105% in trials 7A and 7B, respectively, and by only 64% in trial 7D (with the placebo composition).

[0217] This 7th series of experiments demonstrates that 13-HOD is effective in repairing hair fibers.

[0218] 8 ème Series of experiments: Effect of oil C, used in leave-on cosmetic compositions, on the porosity of the hair cuticle:

[0219] Tests 8a to 8C and the following Control 8 were carried out.

[0220] For each of these trials, 3 strands of hair from a Caucasian donor were used.

[0221] The hair strands in tests 8A to 8C were exposed to heat stress at 95°C for one hour. The strands in Control test 8 were not exposed to such heat stress.

[0222] Test 8A:

[0223] During test 8A, the following cosmetic composition was used. The list of their constituents and their mass percentages, expressed relative to the total mass of the composition, are detailed in Table 4 below. Table 4

[0224] The product was applied to dry hair strands. The hair was then left to air dry. In other words, the hair was not rinsed.

[0225] The hair strands were then soaked in a fluorophore solution (specifically, a fluorescein solution) and cryopreserved. Cross-sections were prepared. The fluorescent signal was visualized by microscopy and quantified by image analysis.

[0226] Test 8B:

[0227] During trial 8B, a cosmetic composition referred to as a "placebo" was used. The list of its constituents and their mass percentages, expressed relative to the total mass of the cosmetic composition referred to as a "placebo," are detailed in Table 5 below. Table 5

[0228] The so-called "placebo" composition was applied to dry hair strands. The hair strands were then left to air dry. In other words, the hair strands were not rinsed.

[0229] The hair strands were then soaked in a fluorescein solution and cryopreserved. Cross-sections were prepared. The fluorescent signal was visualized by microscopy and quantified by image analysis.

[0230] Test 8C:

[0231] No composition was applied to the hair strands in test 8C. The hair strands were washed three times with ultrapure water and then air-dried. They were then soaked in a fluorescein solution and cryopreserved. Cross-sections were prepared. The fluorescent signal was visualized by microscopy and quantified by image analysis.

[0232] Test Control 8:

[0233] The hair strands in Control 8 were not subjected to heat treatment. Furthermore, no cosmetic products were applied to them. The hair strands were washed three times with ultrapure water and then air-dried. Next, they were soaked in a fluorescein solution and then cryopreserved. Cross-sections were prepared. The fluorescent signal was visualized by microscopy and quantified by image analysis.

[0234] Figure 8 is a graph which represents, for tests 8A to 8C and Control 8, the diffusion of the fluorophore within the fibers of the hair strands, said diffusion of the fluorophore being the penetration distance of the fluorophore within said fibers expressed as a percentage relative to the diameter of said fibers.

[0235] In the absence of heat stress, the hair strands in Test Control 8 have fibers whose structural integrity prevents fluorophore diffusion. The fluorophore diffusion rate is 10.78%.

[0236] After heat stress, the fibers of the hair strands became porous (in other words, they were no longer intact) and the diffusion of the fluorophore increased by 88%.

[0237] The higher the diffusion of the fluorophore, the more porous the hair strand fibers become.

[0238] That is why for test 8C, the percentage of diffusion of the fluorophore is 20.25%.

[0239] Treating hair strands with a cosmetic composition containing 13-HOD oil (also known as test 8A) restored the integrity of the hair fibers. The fluorophore diffusion rate was 11.53%.

[0240] Treating the hair strands with a so-called "placebo" composition did not restore the integrity of the hair fibers to the same extent. Indeed, the fluorophore diffusion rate in trial 8B was 13.97%.

[0241] Thus, treating hair strands with a cosmetic composition containing 13-HOD-based oil C restored the integrity of the hair fibers with greater efficacy than the placebo composition. Indeed, fluorophore diffusion decreased by 92% in trial 8A and by only 66% in trial 8B (with the placebo composition).

[0242] This 8th series of experiments demonstrates that 13-HOD is effective in repairing hair fibers.

[0243] Tables 6 to 9 below detail examples of cosmetic compositions according to the invention by listing their constituents and the associated mass percentages, expressed relative to the mass of the cosmetic composition in question. The constituents are listed with their INCI names (INCI being the English abbreviation for "International Nomenclature of Cosmetic Ingredients").

[0244] In all the tables below, the abbreviation "QSP 100" means "quantity sufficient for 100%". This refers to the mass percentage of water required to make the sum of the mass percentages of all the constituents of the cosmetic composition reach 100%.

[0245] All the cosmetic compositions detailed below included C oil.

[0246] Composition of anti-hair loss serum according to the invention: Table 6

[0247] Composition of anti-hair loss shampoo according to the invention: Table 7

[0248] The abbreviation "QSP pH 5 to 6" stands for quantity sufficient for pH from 5 to 6. This is the mass percentage of citric acid required to ensure that the pH of the cosmetic composition is between 5 and 6.

[0249] Composition of hair mask according to the invention: Table 8

[0250] Composition of a nourishing and repairing shampoo according to the invention: Table 9

Claims

DEMANDS 1. Cosmetic composition, characterized in that it comprises at least one modified vegetable oil containing 13-hydroxy-9(Z)-octadecenoic acid (hereinafter abbreviated 13-HOD).

2. Cosmetic composition according to claim 1, characterized in that said modified vegetable oil comprises, in mass percentages expressed in relation to the total mass of free fatty acids contained in said modified vegetable oil, at least 10%, preferably at least 20% and even more preferably at least 40%, of 13-HOD.

3. Cosmetic composition according to claim 1 or 2, characterized in that said vegetable oil is selected from the group consisting of grape seed, safflower, borage, peanut, wheat germ, corn, sunflower, blackberry, nigella, evening primrose, pumpkin seed, hemp, cotton, walnut, soybean, argan and rapeseed oil, taken alone or in mixtures thereof.

4. Cosmetic composition according to claim 3, characterized in that said vegetable oil is a grape seed oil.

5. Cosmetic composition according to any one of claims 1 to 4, characterized in that it comprises, in mass percentages expressed in relation to the total mass of said cosmetic composition, between 0.0001% and 1%, preferably between 0.001% and 0.1%, of 13-HOD.

6. Cosmetic composition according to any one of claims 1 to 5, characterized in that the mass percentage of said modified vegetable oil, expressed in relation to the total mass of said cosmetic composition, is between 0.0005% and 5%, preferably between 0.005% and 0.5%.

7. Non-therapeutic cosmetic treatment method for stimulating hair growth, characterized in that it comprises a step of applying to a determined hairy area a cosmetic composition comprising as an active ingredient 13-HOD.

8. Non-therapeutic cosmetic treatment method for stimulating hair growth according to claim 7, characterized in that said cosmetic composition is a cosmetic composition according to any one of claims 1 to 6.

9. Non-therapeutic cosmetic treatment method according to claim 7 or 8, characterized in that the determined hair area is all or part of the scalp, chin, eyebrows or free edges of the eyelids on which the eyelashes are implanted.

10. Non-therapeutic cosmetic treatment process for hair fiber repair, characterized in that it comprises a step of applying to all or part of the scalp a cosmetic composition comprising 13-HOD as an active ingredient.

11. Non-therapeutic cosmetic treatment process for hair fiber repair according to claim 10, characterized that the cosmetic composition is a cosmetic composition according to any one of claims 1 to 6.

12. Non-therapeutic cosmetic use of 13-HOD to stimulate hair growth.

13. Non-therapeutic cosmetic use of 13-HOD to repair the hair fiber.

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