MXPA00010407A - Assay for identification of compoundsthat promote melanin production and retinoid-like compounds identified by said assay - Google Patents

Abstract

An in vitro assay for selecting compounds that alter pigmentation of skin is provided. Also, a novel class of pro-pigmentatory compounds is provided which comprise substituted aromatic or heterocyclic carboxylic acids, or derivatives thereof, or pharmaceutically acceptable salts, which do not contain a pheno, naphthol, thiophenol, or a thionaphthol function in free or protected form. In a preferred embodiment, thess compounds will display activity for RXRs. These compounds may be used for altering pigmentation of human skin and/or hair in cosmetic or dermatological compositions, and for the treatment of disorders and disease conditions that affect skin or hair pigmentation.

Description

TEST FOR THE IDENTIFICATION OF COMPOUNDS THAT PROMOTE THE PRODUCTION OF MELANINE AND SIMILAR COMPOUNDS TO THE RETINOIDS IDENTIFIED BY THE SESSION Background of the Invention The normal color of the skin is the result of the combined expression of a mixture of different pigments of which melanin, a brown pigment, is the major component. Melanin is synthesized by specialized cells, melanocytes, which are found in the epidermis. Melanocytes synthesize melanin in organelles, which are called melanosomes. As the melanosomes become saturated with melanin) they are transported to the dentritic branches of the melanosomes and are transferred to the surrounding keratinocytes. Then, the keratinocytes migrate to the surface of the skin causing the skin to exhibit a dark pigmentation. The amount of melanin in the keratinocytes determines the extent of pigmentation in the skin and hair.

Because of the effect of melanin on the pigmentation of the skin and hair, investigations have been carried out that aim to identify compounds that affect the production of melanin and / or the Ref. 123425 transport of melanin to the surrounding keratinocytes. Such compounds have potential application for the alteration of the pigmentation of the skin and hair. For example, it is known that the elanocyte stimulating hormone (MSH), and compounds such as theophylline, induce pigmentation. In addition, MSH analogs have been examined for clinical efficiency to promote skin pigmentation. Also, it has been reported that alpha-lipoproteins can stimulate the production of melanin in the skin. (Jpn. Kokai Tokio, Koho, JP 82-153348, GB 2124900, by the Cuban Import and Export Company of Medical Products, Cuba.) Reciprocally, the use of some compound to inhibit pigmentation, for example, is also known. melasma treatment, for example, hydroquinone (HQ), and monobenzene ether (Pathak et al., J. American Academy Dermotol 15: 894-899 (1986); Smith et al., Pigment Cell Research, 1: 386-389 (1988) ), and arbutin a conjugated gluco of HQ (Proceeding Japan Soc. Invest. Dermotol., 12: 138: 139 (1988)).

Furthermore, it has been reported that a kojic acid and derivatives thereof and / or valeric acids inhibit the formation of melanin (Japan Kokai Tokyo Koho, JP 03011010, Oyama, Yasuaki, Sansei Pharmaceutical Co., Ltd, Japan).

Additionally, it has been reported that propionic, butyric, and valeric acids and their salts in combination with or without unsaturated fatty acids suppress the formation of tyrosinase, which catalyzes the production of melanin (Mishima et al., EPA 345081, Hayashibara Biochemical Laboratories , Inc. Japan).

EP 0 509 241 and WO 93/19729 disclose that the hyper-pigmentation, or tanning of the skin, can be carried out by topically applying thereto an efficient amount of trans-retinoic acid for a sufficient time. However, the compounds used in these methods induce irritation.

An in vi tro method that is proposed as adequate to identify compounds that de novo inhibit the synthesis of melanin and potential anti-melanoma, cytotoxic compounds was reported by Dooley et al., Pharmacol. , 7: 188-200 (1994). These compounds use two immortalized murine cell lines of melanocyte origin, Mel-Ab (Dooley et al., Oncogene, 3: 531-535 (1988)), and the other of fibroblast origin. Mel-Ab cells are derived from a spontaneously immortalized murine C57BL \ 6 melanocyte line that reproduces rapidly in culture and produces copious amounts of melanin. This assay, according to the reports received, was used to select compounds that have structure similar to intermediaries in the biosynthetic pathway of melanin that are unique for melanocytes. In particular, para-substituted phenols related to tyrosine and 1, 2-dihydrobenzene related to DOPA were selected, as well as 1, 3-dihydrobenzenes, 1,4-dihydroxybenzenes, 1,4-dimethoxybenzenes, and 1,4-benzoquinones. Several compounds that reportedly alter pigmentation were identified using this selection. However, unlike the present invention this in vi tro selection preferably uses immortalized murine cell lines and not normal human melanocytes.

However, it would be beneficial if other compounds that carry out the pigmentation and production and / or transport of melanin were identified. It would also be beneficial if compounds that are well tolerated and not irritating to the skin were identified. In addition, it would be useful if an in vi tro selection test were developed to identify such compounds.

Brief Description and Objectives of the Invention For this purpose, it is an object of this invention to provide a new assay for the identification of compounds that can alter (improve or inhibit) the expression of melanin by melanocytes, preferably human melanocytes.

It is another object of the invention to provide pharmaceutical / dermatological / cosmetic compositions comprising inhibitory or promoter amounts of melanin, of at least one compound identified by such assay.

It is a more specific objective of the invention to provide pharmaceutical / cosmetic / dermatologically applicable compositions comprising melanin inhibiting or enhancing amounts of at least one compound that alters (improves or inhibits) the production of melanin by human melanocytes.

It is also a more specific object of the invention to provide dermatological / cosmetic / pharmaceutical compositions comprising an enhancing or inhibiting amount of melanin, of at least one substituted heterocyclic or aromatic carboxylic acid or a derivative thereof with the proviso that said carboxylic acid does not comprises a functional group free phenol, naphthol, thiophenol, or thionephthol, or one which is protected by a protecting group.

It is another specific object of the invention to provide a method of altering the pigmentation of the skin and / or hair in a subject in need of such treatment, comprising topically applying an effective amount of a composition comprising at least one retinoid compound that affects to melanin (inhibitor or that increases), for a sufficient time to induce pigmentation of the skin and / or hair.

It is a more specific objective of the invention to provide a method of alteration of either increasing or decreasing the pigmentation of the skin and / or of the hair, in a subject in need of such treatment, comprising topically applying an effective amount of a composition. comprising at least one heterocyclic or substituted aromatic carboxylic acid or a derivative thereof for a time sufficient to affect the pigmentation of the skin and / or hair, provided that said carboxylic acid does not comprise a phenol, naphthol, thiophenol group , or tionaphthol, free or protected.

Brief description of the figures Figure 1 contains the structures of the compounds identified according to the invention that affect the production of melanin; Y Figures 2 through 4 list pigmentation diseases that can be treated using the compounds of the invention.

Detailed description of the invention In a first embodiment, the present invention provides a new assay for identifying compounds that alter (increase or inhibit) the expression of melanin by human melanocytes, preferably compounds that increase the production of melanin by human melanocytes.

In general, the test content will test the effect of a particular compound or combination of compounds on the growth or proliferation of human melanocytes in culture, and also the effect of said compound on the production of melanin by said cultured human melanocytes, normalizing the Melanin content based on the number (proliferation) of said cultured human melanocytes, and will identify a compound or compounds that affect the production of melanin based on an increase or decrease in production per cell, of melanin.

More specifically, the development of the trial will include: (i) contacting at least two cultures of human melanocytes with at least one compound to be selected for this effect on melanin production; (ii) measuring the proliferation of melanocytes contained in one of said human melanocyte cultures; (iii) measuring the amount of melanin contained in a second culture of human melanocytes; (iv) compare the data obtained in steps (ii) and (iii) in order to determine the effect of such compound or compounds on the production of melanin (increase or decrease), and normalize the data based on the number of melanocytes contained in said crops; (v) further comparing the data obtained in steps (ii) and (iii) with the proliferation and production of melanin in control cultures of human melanocytes that are cultured under conditions identical to those of the above cultures except in the absence of said compound; Y (vi) identifying a compound selected as one that alters (increases or decreases) the expression of melanin by human melanocytes based on an increase or decrease in the production of melanin by melanocytes cultured in the presence of said compound in relation to control cultures not contacted with said compound.

This method will preferably be effected by obtaining human melanocytes, for example, neonatal skin melanocytes, from a suitable source. These melanocytes are then seeded simultaneously in two cell culture chambers, for example, a 96-well plate and a 24-well plate. After such cells were allowed to adhere to the surface of the wells, the respective wells are then incubated with a fixed amount of a particular compound or compounds, whose effect on the expression of melanin is to be determined. Such incubation will be effected for a sufficient period of time to evaluate the effect, if any, of such compound or compounds on the production of melanin by such cultured melanocytes.

The incubation will then vary from about one minute to about one month, more preferably from about one hour to about two weeks, and more preferably from about 12 hours to about a week. After incubation, the proliferation of the cells in one of said cultures is then evaluated. This can be effected by suitable methods, for example, by absorbance.

In a preferred embodiment, the cells in the 96 wells are treated with 3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium (a tincture of tetrazolium conventionally used to determine colorimetrically the proliferation of the cells) (MTS) and the absorbance of the plate read at 490 nm to determine the proliferation of the cells in said 9-well plate. These results are then compared with those of a melanocyte control culture that was cultured under the same conditions, and subsequently treated with MTS solution, but not treated with the test compound.

The second culture, for example, a 24-well plate, containing a culture of human melanocytes which was incubated with the test compound under the same conditions, is then examined to determine the amount of melanin expressed by the cells contained in the same. This can be effected by known methods. In the present invention, this will preferably be effected by washing such cells, for example, with regulatory saline (PBS), using said washed cells with an alkaline solution, for example, with IN NaOH, and measuring the amount of soluble melanin in alkali contained therein, for example, by measurement of the absorbance of alkali-soluble melanin at 45 nm. This data is preferably compared to a control culture of melanocytes, developed under the same conditions but in the absence of said test compound.

The data obtained from both cultures of melanocyte cells is then normalized based on the proliferation of melanocytes (number of cells) in order to determine the effect, if any, of the particular compound or compounds on the production and accumulation of melanin by the melanocytes.

The compounds that alter the expression of melanin according to the invention are those compounds that affect the production and accumulation of melanin in cultured human melanocytes. Preferably, such a compound will alter the expression of melanin by at least 10% in relation to the control cultures of melanocytes. More preferably such a compound will affect the production of melanin by at least 20%. More preferably, the compound will affect the production and accumulation of melanin in the order of 150 to 250%, or greater.

Accordingly, the content of the invention is directed towards the identification of compounds or combinations thereof that increase or decrease the production and accumulation of melanin of human melanocytes. Compounds that increase the production and accumulation of melanin can be used, for example, to promote tanning or darkening of the skin and to treat or prevent shrinking of the hair. Also, such compounds can be used to treat diseases or conditions associated with hypo-pigmentation, for example, vitiligo.

Compounds that decrease the production of melanin can be used for the treatment of subjects having diseases or conditions associated with hyper-pigmentation. Such diseases and conditions include melasma or hyper-pigmentation related to age, and chloasma.

Also, compounds that inhibit the production and / or accumulation of melanin can be used to treat and / or prevent hyper-pigmentation associated with age, for example, "liver spots" often observed on the hands and face of elderly people. A list of diseases and conditions associated with pigmentation disorders can be found in Dermatology in General Medicine (Fitzpatric, T. B., et al.) Which is incorporated herein by reference in its entirety.

The compounds of the present invention can be useful in the treatment of hypo-pigmentation, applications such as vitiligo, hypo-pigmentations or post-inflammatory depigmentation, for the treatment of hypo-pigmentations or depigmentation after skin grafts, for the treatment of hypo-pigmentations or post-cicatrization depigmentation, or for the treatment of hypo-pigmentations or depigmentation due to old age or lentigo.

Compounds that decrease the production of melanin may be useful in the treatment of melasma or hyper-pigmentation associated with age.

Additionally, the content of the assays can be used to identify combinations of compounds that affect (increase or decrease) the production or accumulation of melanin by human melanocytes, in particular combinations that have synergistic effects on the production or accumulation of melanin.

As discussed above, in a second embodiment, the present invention is directed toward pharmaceutical / dermatological / cosmetic compositions containing at least one compound that affects the melanin according to the invention. In this regard, a new class of compounds has been discovered using the content of assay methods that promote the production and / or accumulation of melanin by normal human melanocytes.

These compounds are also advantageous in that they are well tolerated and do not irritate the skin. This is surprising because many other retinoid compounds selected using the assay content do not affect pigmentation under similar conditions. These compounds hypothetically affect the production and / or accumulation of melanin via the retinoid indicator method.

In a preferred embodiment, the present invention provides pharmaceutical / cosmetic / dermatological compounds comprising an amount of at least one substituted heterocyclic or aromatic carboxylic acid, or a derivative thereof, with the proviso that such carboxylic acid does not contain a phenol function, naphthol, thiophenol, or tionephthol, in free or protected form.

Such a carboxylic acid includes, by way of example, benzoic acid, propiolic acid, nicotinic acid, acrylic acid, butenoic acid and naphthoic acid.

In a preferred embodiment, the substituted heterocyclic or aromatic carboxylic acids of the present invention will exhibit activity by RXRs. According to the present invention, a compound exhibiting RXR activity is a compound that exhibits an RXR bond of less than 5000 nM, and preferably less than 1000 nM. The determination of the linkage by RXRs is well known to those skilled in the art and is reported in, for example: (1) "Selective Synthetic Ligands for Nuclear Retinoic Acid Receptor Subtypes" in RETINOIDS, Progress in Research and Clinical Applications, Chapter 10 ( pp 261-267), Marcel Dekker Inc., published by Maria A. Livrea and Lester Packer; (2) Synthetic Retinoids: Receptor Selectivity and Biological Activity "in Pharmacol Skin, Basel, Karger, 1993, Vol. 5 pp 117-127; (3)" Selective Synthetic Ligands for Human Nuclear Retinoic Acid Receptors "in Skin Pharmacology, 1992, Vol. 5, pp 57-65; (4) "Identification of Synthetic Retinoids with Selectivity for Human Nuclear Retinoic Acid Receptor-?" In Biochemical and Biophysical Research Communications, Vol. 186, No. 2, July 1992, pp. 977- 983; and (5) "Selective High Affinity RAR-a or RAR-ß Retinoic Acid Receptor Ligands" in Mol.Pharmacol., Vol. 40, pp. 556-562.

Also, in a more preferred embodiment, the compound exhibiting activity by RXRs, exhibits an agonist activity by RXRs. This agonist activity by RXRs can be determined for example by the method reported in US Patent No. 5,696,104, the total content of which is incorporated herein by reference.

More specifically, compounds that have been surprisingly discovered to promote the production and / or accumulation of melanin by the human melanocytes according to the invention include: 4- [1 - (5, 6, 7, 8-Tetrahydro-5, 5, 8, 8-tetramethyl-2-naphthyl) ethenyl] benzoic acid (CD No. 2771); 3- [3- (5,5,8,8-Tetramethyl-5,6,7,8-tetrahydro-naphthalen-2-yl) -phenyl] -acrylic acid (CD No. 2908); 3- [3- (3, 5, 5, 8, 8-pentamethyl-5, 6,7,8-tetrahydro-naphthalen-2-yl) -phenyl] -but-2-enoic acid (CD No. 3206); 6- [I- (3, 5, 5, 8, 8-pentamethyl-5, 6, 7, 8-tetrahydra-naphthalen-2-yl) -cyclopropyl] -nicotinic acid (CD No. 3127); 3- (3-, 3, 5, 5, 8, 8-pentamethyl-5,6,7,8-tetrahydro-naphthalen-2-yl) -phenyl] -acrylic acid (CD No. 2915); 4- (3, 5, 5, 8, 8-pentamethyl-5, 6, 7, 8-tetrahydro-naphthalene-2-carbonyl) -benzoic acid (CD No. 2608); 4- (3-Bromo-5, 5, 8, 8-tetramethyl-5, 6, 7, 8-tetrahydro-naphthalen-2-yloxy) -benzoic acid (CD No. 2661); - [(E) -2- (3, 5, 5, 8, 8-pentamethyl-5,6,7,8-tetrahydro-naphthalen-2-yl) -propenyl] -thiophen-3-carboxylic acid (CD No. 2425); 3- [3- (5, 5, 8, 8-tetramethyl-5, 6,7,8-tetrahydro-naphthalen-2-ylsulfanyl) -phenyl] -acrylic acid (CD No. 3132); 6- (3-Butyl-5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-naphthalen-2-ylsulfanyl) -nicotinic acid (CD No. 3292); 4- (3, 5, 5, 8, 8-pentamethyl-5, 6, 7, 8-tetrahydro-2-naphthylthio) benzoic acid (CD No. 2624); 3- (3, 5, 5, 8, 8-pentamethyl-5,6,7,8-tetarhydro-2-naphthyl) phenylpropiol acid (CD No. 2906); Y 6- (3, 5, 5, 8, 8-pentamethyl-5,6,7,8-tetrahydro-2-naphthylthio) nicotinic acid (CD No. 2809).

As noted above, the pharmaceutical / dermatological / cosmetic compositions will be in a form suitable for topical application to the skin and / or hair. In a preferred embodiment, these compositions will be in a form suitable for artificial tanning and / or darkening of the skin.

These compositions may comprise other pharmaceutically, cosmetically or dermatologically acceptable constituents typically formulated in these types of compositions, such as thickening agents, softeners, antioxidants, opacifiers, stabilizers, emollients, organic sunscreens which are active in the UV-A regions and / or UV-B, inorganic photoprotective pigments, and non-pigments, humidifiers, vitamins, fragrances, preservatives, filters, sequestrants, anilines, and dyes.

Naturally, one skilled in the art would be careful in selecting this or these optimal complementary compounds and / or the amounts thereof which advantageous properties intrinsically associated with the compositions of the invention are not adversely affected by the addition or additions considered.

The inventive compositions may comprise one or more compounds that affect the production and / or accumulation of melanin by the melanocytes, for example, synergistic combinations. Also, such compositions may comprise other compounds that promote or inhibit pigmentation, for example dihydroxyacetone (DHA), or derivatives thereof. Suitable compounds are described in Serial No. USA 08 / 819,084, registered on March 18, 1997 by Tuloup et al., And assigned to L'OREAL. Also, M? H and analogs thereof that promote pigmentation may be included. Reciprocally, compounds that inhibit pigmentation such as HQ, monobenzene ether and arbutin can be included with compounds identified using the content of the assay which inhibit pigmentation.

The compositions contained herein will be in the form suitable for topical applications on skin and / or human hair. Suitable forms include a cream, milk, a gel-cream, a fluid lotion, an oil-in-water emulsion or water-in-oil emulsion, a vesicle dispersion, or any other form typically employed in the art.

The amount of at least one compound that affects the melanin contained in the compositions herein will typically range from about 0.0001% to about 30% by weight relative to the total weight of the composition, and preferably will vary in concentration from about 0.5% to about 15% by weight relative to the total weight of the composition, and more preferably will vary in concentration from about 0.001% to about 5% by weight relative to the total weight of the composition.

The present invention also encompasses a treatment, cosmetic, dermatological, or therapeutic regimen, comprising the topical application of an amount of a compound that affects the melanin according to the invention.

An effective amount is an amount sufficient to affect (increase or decrease) the pigmentation of the skin and / or hair. Such a regimen may be carried out in conjunction with other compounds that affect the pigmentation of the skin and / or hair. Such a regimen can be effected repeatedly until the desired effect on pigmentation is achieved. Chronic administration would be suitable when the compounds of the present invention are not irritating to the skin.

As discussed, the present invention encompasses, in particular, compositions containing at least one compound according to the invention which are suitable for promoting hair coloring. Such compositions may include other constituents typically included in hair formulations, eg, conditioning agents, pigments and color ingredients, spreaders, perms, surfactants, perfumes, alcohols, etc.

In order to facilitate a better understanding of the invention, the following examples are provided.

EXAMPLE 1 Normal human neonatal skin melanocytes were used for this assay. For primary selection, the cells were seeded simultaneously into 96-well plates and approximately 24-well plates according to the surface area of the wells, and were treated with the test compound for 5 days. The addition of the compound to the plates was done using a Robot, to ensure accuracy (same amount in each well). At the end of the five days of treatment, the cells of the 96-well plate were treated with MTS solution and the plate was read at 490 nm to determine the effect of the compounds on the growth of the cells. The cells in the 24-well plates were rinsed with PBS, and then used with IN NaOH, and the absorbance of the alkali-soluble melanin was measured at 405 nm.

Data from the 96-well plate were analyzed to determine the effect of each compound on cell proliferation, and the 24-well plate data was used to determine the effect of the compound on the melanin content of the cells. The melanin content was then normalized to the proliferation of the cells.

Using this method, the inventors identified the following compounds that increase the melanin content of normal human melanocytes. It is anticipated based on these results, that other compounds can be identified, for example, retinoids, which alter (increase or decrease) the production of melanin by human melanocytes.

CD No, Maximum Increase in Observations Melanin Content (Normalized) 2771 250% at 2 μM 2908 200% at 2 μM 3206 198% at 0.5 μM 2624 181% at 2 μM 3127 179% at 0.1 μM 2906 171% at 0.1 μM 2915 165% to 0.1 μM CD No. Maximum increase in Observations Melanin Content (Normalized) 2608 157% at 2 μM 2661 155% at 1 μM 2425 149% at 1 μm 2809 145% at 0.5 μM 3132 142% at 2 μM 3292 121% of 1 μM Thus, using the assay of the present invention, the inventors have identified a new class of pro-pigment compounds for normal human melanocytes (thus increasing the production of melanin). It is hypothetical that these molecules can work through the retinoid indicator pathway. The discovery of a class of molecules similar to retinoids that have pro-pigment activity is surprising especially because other retinoid indicator molecules do not induce pigmentation under similar conditions. This discovery is additionally advantageous because these compounds are very well tolerated, and are not skin irritants. Consequently, they have potential application in the alteration of pigmentation in the skin and / or hair of human subjects.

EXAMPLE 2 EXAMPLES OF FORMULATIONS 1. ORALLY (a) The following composition is prepared in the form of a 0.8 g tablet: Compound of formula (2) 0.005 g 0.265 g pre-gelatinized starch Microcrystalline cellulose 0.300 g Lactose 0.200 g Magnesium Stearate 0.030 g For the treatment of post-inflammatory hypo-pigmentation, 1 to 3 tablets are administered to an adult individual per day for three to six months depending on the severity of the case treated. (b) An ingestible suspension is prepared which was planned for packaging in 5 ml vials, Compound of formula (4) 0.050 g Glicerol 0.500 g Sorbitol 70% 0.500 g Sodium saccharinate 0.010 g Methyl para-hydroxybenzoate 0.040 g Taste enough quantity Purified water c.b.p. 5,000 ml For the treatment of a hypo-pigmentation after a skin graft, a vial is administered to an adult individual per day for three months depending on the severity of the case treated. (c) The following formulation that was planned for packaging in gelatin capsules was prepared: Compound of formula (5) 0.025 g Cornstarch 0.060 g Lactose c.b.p. 0.300 g The gelatin capsules used consist of gelatin, titanium dioxide, and a preservative agent.

In the treatment of vitiligo, a gelatin capsule was administered to an adult individual per day for 30 days. 2. VIA TÓPICA (a) The following non-ionic-in-oil water cream was prepared: Compound of formula (8) 0.100 g Mixture of lanolin alcohols emulsifiers, waxes and refined oils, sold by the company BDF under the name "Eucerin anhydrous" 39,900 g Methyl para-hydroxybenzoate 0.075 g Propyl para-hydroxybenzoate 0.075 g Sterile demineralized water c.b.p. 100,000 g This cream is applied to a hypo-pigmented skin once or twice a day for thirty days. (b) A gel was prepared by preparing the following formulations: Compound of formula (11) 0.050 g Erythromycin base 4000 g Butylhydroxytoluene 0.050 g Hydroxypropyl cellulose sold by the company Hercules under the name of "Klucel HF" 2,000 g Ethanol (95%) c.b.p. 100,000 g This gel is applied to a hypo-pigmented grafted skin one to three times a day for six to twenty weeks depending on the severity of the treated case. (c) A lotion was prepared for correction of post-inflammatory hypo-pigmentation, by mixing together the following ingredients: Compound of formula (2) 0.030 g Propylene glycol 5,000 g Butylhydroxy toluene 0.100 g Ethanol (95 °) c.b.p. 100,000 g This lotion is applied twice a day and a significant improvement is observed in a period of two to six weeks. (d) A cosmetic composition is prepared to combat the damaging effect of the sun's rays by mixing together the following ingredients: Compounds of formula (4) 1,000 g Benzylidene camphorated 4000 g Triglycerides of fatty acids 31,000 g Glyceryl monostearate 6,000 g Stearic acid 2,000 g Cetilic alcohol 1,200 g Lanolin 4000 g Conservative agents 0.300 g Propylene glycol 2,000 g Triethanolamine 0.500 g Fragrance 0.400 g Demineralized water c.b.p. 100,000 g This composition is applied daily and makes it possible to combat the aging induced by light. (e) The following cream was prepared with non-ionic-in-water oil: Compound of formula (3) 0.500 g Vitamin D3 0.020 g Cetilic alcohol 4,000 g Glyceryl monostearate 2,500 g Stearate PEG-50 2,500 g Butter Karite 9.200 g Propylene glycol 2,000 g Methyl para-hydroxybenzoate 0.075 g Propyl para-hydroxybenzoate 0.075 g Sterile demineralized water c.b.p. 100,000 g This cream is applied to a skin affected with vitiligo once or twice a day for thirty days. (f) A topical gel was prepared by mixing together the following ingredients: Compound of formula (17) 0.050 g Ethanol 43,000 g a-tocopherol 0.050 g Carboxyvinyl polymer sold under the name Carbopol 941® by "Goodrich" 0.500 g Triethanolamine 20% by weight in aqueous solution 3,800 g Water 9,300 g Propylene glycol c.b.p. 100.00 g This gel is applied to a skin affected with vitiligo one to three times a day for six to twelve weeks depending on the severity of the case treated. (g) A hair lotion was prepared to re-pigment the hair by mixing together the following ingredients: Compound of formula (9) 0.05 g Compound sold under the name of Minoxidil® 1.00 g Propylene glycol 20.00 g Ethanol 34.92 g Polyethylene glycol (molecular mass = 400) 40.00 g Butylhydroxyanisole 0.01 g Butylhydroxytoluene 0.02 g Water c.b.p. 100,000 g This lotion is applied twice a day for three months to a scalp that has undergone considerable depigmentation. (h) A post-healing cream was prepared by mixing together the following ingredients: Compound of formula (13) 0.050 g Retinoic acid 0.010 g Mixture of glyceryl stearate and polyethylene glycol stearate (70 moles) sold under the name of Gelot 64® by "Gattefosse" 15,000 g Polyoxyethylenated corn grain oil with 6 moles of ethylene oxide, sold under the name Labrafil M2130 CS®, for "Gattefosse" 8,000 g Perhydrosqualene 10,000 g Polyethylene glycol (molecular mass = 400) 8,000 g Ethylenedia inotetraacetic acid disodium salt 0.050 g Purified water c.b.p. 100,000 g This cream is applied one to three times a day for six to twelve weeks. (i) An oil-in-water cream was prepared by making the following formulation: Compound of formula (4) 0.020 g Betamethasone 17-valerate 0.050 g S-carboxymethylcysteine 3,000 g Polyoxyethylene stearate (40 moles of ethylene oxide) sold under the name of Myrj 52® by "Atlas" 4,000 g Sorbitan monolaurate, polyoxyethylenated with 20 ml of ethylene oxide, sold under the name of Tween 20® by "Atlas" 1,800 g Mixture of glyceryl mono- and di-stearate sold under the name of Galeol®, by "Gattefosse" 4.200 g Propylene glycol 10,000 g Butylhydroxyanisole 0.010 g Butylhydroxytoluene 0.020 g Cetostearyl alcohol 6,200 g Sufficient amount of preservatives) perhydrosqualene 18,000 g Caprylic / capric triglyceride mixture sold under the name Miglyol 812® by "Dynamit Nobel" 4,000 g Triethanolamine (99% by weight) 2,500 g Water c.b.p. 100,000 g This cream is applied twice a day to an affected skin with pigmentation problems due to aging. (j) The following oil-in-water type cream was prepared: Lactic acid 5,000 g Compound of formula (13) 0.020 g Polyoxyethylene stearate (40 moles of ethylene oxide) sold under the name Myrj 52® by "Atlas" 4,000 g Sorbitan monolaurate, polyoxyethylenated with 20 moles of ethylene oxide sold under the name of Tween 20®, by "Atlas" 1,800 g Mixture of glyceryl mono- and di-stearate sold under the name of Celeol®, by "Gattefosse" 4.200 g Propylene glycol 10,000 g Butylhydroxyanisole 0.010 g Butylhydroxytoluene 0.020 g Cetostearyl alcohol 6,200 g Conservative agents (sufficient amount) Perhydrosqualene 18,000 g Mixture of triglycerides aprilico / caprico sold under the name Miglyol 812® by "Dynamit Nobel" 4.000 g Water c.b.p. 100,000 g (k) Dermal lotion for aerosol Compound of formula (15) 5,000 g Ethanol 30,000 g Demineralized water c.b.p. 100,000 g (1) Hair lotion: Compound of formula (18) 3,000 g Propylene glycol 30,000 g Ethyl alcohol 40,500 g Water c.b.p. 100,000 g This lotion is applied on the scalp one or eces a day in a proportion of 1 ml per application. (m) Thick lotion: Compound of formula (1) 5,000 g Kawaine 2,000 g Hydroxypropylcellulose (Klucel G® from Hercules) 3,500 g Ethyl alcohol c.b.p. 100,000 g This thick lotion is applied to the scalp once or twice a day in a proportion of 1 ml per application. (n) Niosomic lotion Chimexane ML® 0.475 g Cholesterol 0.475 g Monosodium stearoyl glutamate 0.050 g Compound of formula (3) 0.100 g Conservative agents sufficient amount Enough quantity Enough quantity Demineralized water c.b.p. 100,000 g This lotion is applied to the scalp once or twice a day in a proportion of 1 ml per application. (o) Lotion: Compound of formula (17) 5,000 g Propylene glycol monomethyl ether (Dowanol PM® from Dow Chemical) 20,000 g Hydroxypropylcellulose (Klucel G® from Hercules) 3,000 g Ethyl alcohol 40,000 g Minoxidil 2,000 g Water c.b.p. 100,000 g This thick lotion is applied to the scalp once or twice a day in a proportion of 1 ml per application.

While the invention has been described with respect to certain specific embodiments, it will be appreciated that those skilled in the art make many modifications and changes thereto without departing from the spirit of the invention. It is therefore intended, with the appended claims, to cover all modifications and changes that fall within the true spirit and scope of the invention.

It is noted that in relation to this date the best method known by the applicant to carry out the aforementioned invention is that which is clear from the present description of the invention.

Having described the invention as above, it is claimed as property in the following:

Claims (28)

1. A cosmetic, dermatological, and / or therapeutic method for altering (promoting or inhibiting) the pigmentation of human skin and / or hair of a human subject, which is characterized in that it comprises applying topically an effective amount of at least one heterocyclic carboxylic acid or substituted aromatic, derivative or pharmaceutically acceptable salt thereof for a period of time sufficient to induce pigmentation in the skin and / or hair, with the proviso that said carboxylic acid does not comprise a phenol, naphthol, thiophenol, or tionephthol function , either free or protected by a protective group.

2. The method of claim 1, characterized in that said compound is a retinoid indicator molecule that promotes pigmentation of the skin and / or human hair.

3. The method of claim 1, which is characterized in that said compound is selected from the group consisting of: 4- [1 - (5, 6, 7, 8-tetrahydro-5, 5, 8, 8-tetramethyl-2-naphthyl) ethenyl] benzoic acid; 3- [3- (5, 5, 8, 8-tetramethyl-5, 6, 7, 8-tetrahydro-naphthalen-2-yl) -phenyl] -acrylic acid; 3- [3- (3, 5, 5, 8, 8, -pentamethyl-5, 6, 7, 8-tetrahydro-naphthalen-2-yl) -phenyl] -but-2-enoic acid; 6- [1- (3,5,5,8,8-pentamethyl-5,6,7,8-tetrahydro-naphthalen-2-yl) -cyclopropyl] -nicotinic acid; 3- [3- (3, 5, 5, 8, 8-pentamethyl-5,6,7,8-tetrahydro-naphthalen-2-yl) -phenyl] -acrylic acid; 4- (3, 5, 5, 8, 8-pentamethyl-5,6,7,8-tetrahydro-naphthalene-2-carbonyl) -benzoic acid; 4- (3-Bromo-5, 5, 8, 8-tetramethyl-5, 6, 7, 8-tetrahydro-naphthalen-2-yloxy) -benzoic acid; 5 - [(E) -2 (3,5, 5, 8, 8-pentamethyl-5,6,7,8-tetrahydro-naphthalen-2-yl) -propeny1] -thiophene-3-carboxylic acid; 3- [3- (5, 5, 8, 8-tetramethyl-5, 6, 7, 8-tetrahydro-naphthalen-2-ylsulfanyl) -phenyl] -acrylic acid; 6- (3-Butyl-5, 5, 8, 8-tetramethyl-5, 6, 7, 8-tetrahydro-naphthalen-2-ylsulfañyl) -nicotinic acid; 4- (3,5, 5, 8, 8-pentamethyl-5,6,7,8-tetrahydro-2-naphthylthio) benzoic acid; 3- (3, 5, 5, 8, 8-pentamethyl-5,6,7,8-tetrahydro-2-naphthyl) phenyl propionic acid; Y Acid 6- (3, 5, 5, 8, 8-pentamethyl-5, 6, 7, 8-tetrahydro-2-naphthylthio) nicotinic acid.

4. The method of claim 1, characterized in that it is used to promote darkening or tanning of human skin;

5. The method of claim 1, which is characterized in that it is used to promote the coloring of human hair.

6. The method of claim 1, characterized in that said compound is contained in a topically applicable composition comprising from 0.001 to 30% by weight of said compound in relation to the weight of the composition.

7. The method of claim 1, characterized in that the amount of said compounds varies from 0.5 to 15% by weight of the composition.

8. A pharmaceutical / cosmetic / dermatological composition which is characterized in that it alters the pigmentation of the skin and / or human hair by topical application, comprising (i) an amount of at least one substituted heterocyclic or aromatic carboxylic compound, derivative, or pharmaceutically acceptable salt thereof, with the proviso that said compound does not contain a free or protected phenol, naphthol, thiophenol or tionaphtol group that is effective for alter pigmentation or skin and / or human hair; Y (ii) a pharmaceutical, cosmetic, or dermatologically acceptable vehicle.

9. The composition of claim 8, which is characterized in that said compound is selected from the group consisting of: 4- [1 - (5, 6, 7, 8-tetrahydro-5, 5, 8, 8-tetramethyl-2-naphthyl) ethenyl] benzoic acid; 3- [3- (5, 55, 8, 8-tetramethyl-5,6,7,8-tetrahydro-naphthalen-2-yl) -phenyl] -acrylic acid; 3- [3- (3, 5, 5, 8, 8, -pentamethyl-5, 6, 7, 8-tetrahydro-naphthalen-2-yl) -phenyl] -but-2-enoic acid; 6- [1- (3, 5, 5, 8, 8-pentamethyl-5,6,7,8-tetrahydro-naphthalen-2-yl) -cyclopropyl] -nicotinic acid; 3- (3- (3, 5, 5, 8, 8-pentamethyl-5,6,7,8-tetrahydro-naphthalen-2-yl) -phenyl] -acrylic acid; 4- (3, 5, 5, 8, 8-pe.tamstil-5, 6, 7, 8-tetrahydro-naphthalene-2-carbonyl) -benzoic acid; 4- (3-Bromo-5, 5, 8, 8-tetramethyl-5,6,7,8-tetrahydro-naphthalen-2-yloxy) -benzoic acid; 5- [(E) -2- (3, 5, 5, 8, 8-pentamethyl-5,6, 1,8-tetrahydro-naphthalen-2-yl) propenyl] -thiophene-3-carboxylic acid; 3- [3- (5,5,8,8-Tetramethyl-5,6,7,8-tetrahydro-naphthalen-2-ylsulfanyl) -phenyl] -acrylic acid; 6- (3-Butyl-5, 5, 8, 8-tetramethyl-5, 6, 7, 8-tetrahydro-naphthalen-2-ylsulfanyl) -nicotinic acid; 4- (3,5,5,8,8-pentamethyl-5,6,7,8-tetrahydro-2-naphthylthio) benzoic acid; 3- (3,5,5,8,8-pentamethyl-5,6,7,8-tetrahydro-2-naphthyl) phenyl propionic acid; Y Acid 6 (3,5,5,8,8-pentamethyl-5,6,7,8-tetrahydro-2-naphthylthio) nicotinic acid.

10. The composition of claim 8, characterized in that the amount of said compound varies from 0.001 to 30% by weight in relation to the weight of the composition.

11. The composition of claim 8, characterized in that the amount of said compound varies from 0.5 to 15% by weight in relation to the weight of the composition.

12. The composition of claim 8, which is characterized in that it is in a form suitable for topical application selected from the group consisting of a cream, a milk, a cream-gel, a fluid lotion, an oil-in-water emulsion, an emulsion water-in-oil, and a dispersion in a vesicle.

13. The composition of claim 8, characterized in that it comprises another compound that affects the pigmentation of the skin and / or the hair.

14. The composition of claim 8, characterized in that it additionally comprises at least one substituent selected from the group consisting of ionic or non-ionic thickeners, softeners, antioxidants, opacifiers, stabilizers, emollients, organic sunblock that are active in the UV region -A and / or UV-B, inorganic photoprotectors and nano-pigments, humidifiers, vitamins, fragrances, preservatives, fillers, sequestering agents, anilines, and dyes.

15. An in vi tro method for selecting a compound or combination of compounds that alters (increases or decreases) the pigmentation of the skin and / or human hair that is characterized in that it comprises the following steps: (i) incubating at least two melanocyte cultures with a compound or combination of compounds, whose effect on the pigmentation of the skin and / or human hair is to be evaluated; (ii) measuring the effect of the compound or combination of compounds mentioned, on the proliferation of melanocytes contained in one of said cultures in relation to a melanocyte control culture that has been cultivated under the same conditions as the mentioned culture except for the absence of the mentioned compound or combination of compounds; (iii) simultaneously or substantially simultaneously with step (ii), measuring the melanin content of cells contained in the second of said cultures that have been incubated with the said compound or combination of compounds and comparing said melanin content to a culture control developed under the same conditions, but in the absence of the mentioned compound or combination of compounds; (iv) compare the data obtained in stages (ii) and (iii) and normalize said data based on the proliferation (number of cells) in said melanocyte cultures in order to determine the effect, if any, of said compound or combination of compounds, on the production of melanin by the melanocytes.

16. The method of claim 15, which is characterized in that said melanocyte cultures comprise human melanocytes.

17. The method of claim 16, which is characterized in that said human melanocytes are obtained from neonatal skin.

18. The method of claim 15, characterized in that step (ii) comprises determining the proliferation of melanocytes in said first culture by absorbance.

19. The method of claim 18, which is characterized in that step (ii) comprises treating said cells with a solution of (MTS) 3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2 - (4-sulfonyl) -2H-tetrazolium and determine the absorbance of said cells treated with MTS at 490 nm.

20. The method of claim 15, wherein in step (iii) the melanin content is determined by lysate of the cells, treatment of the cells with an alkaline solution, and measurement of the absorbance at 405 nm to determine the amount of melanin soluble in alkali contained in it.

21. The method of claim 15, characterized in that said compound or combination of compounds is selected on the basis that it alters the melanin content by at least 20% relative to a melanocyte control culture that is not treated with said compound or combination of compounds.

22. The method of claim 21, characterized in that said compound or combination of compounds alters the melanin content relative to a control melanocyte culture by at least 50%.

23. The method of claim 22, which is characterized in that said compound or combination of compounds alters the melanin content in relation to a control culture of melanocytes by at least 150%.

24. The method of claim 22, which is characterized in that said compound or combination of compounds alters the melanin content in at least 250

25. The method of claim 1, which is characterized in that said compound exhibits activity for RXRs.

26. The composition of claim 8, which is characterized in that said compound exhibits activity for RXRs.

27. The method of claim 25, which is characterized in that said compound exhibits a binding to RXR of less than 5000 nM.

28. The composition of claim 26, which is characterized in that said compound exhibits a linkage for RXR of less than 5000 n.