COSMETIC COMPOSITION COMPRISING COLEUS FORSKOHLII AND CASSIA OCCIDENTALIS AND/OR CASSIA ALATA, AND COMPOSITIONS FOR USE IN TREATING VITILIGO

Abstract

A topical composition comprising: a) a cosmeceutically acceptable vehicle; b) an extract of Coleus forskohlii; and c) an extract of Cassia occidentalis and/or Cassia alata.

Claims

1. A topical composition comprising: a) a cosmeceutically acceptable vehicle; b) an extract of Coleus forskohlii; and c) an extract of Cassia occidentalis and/or Cassia alata.

2. A topical composition according to claim 1, characterized in that it further comprises a penetration enhancer.

3. A topical composition according to claim 1, wherein the extract of Coleus forskohlii is a root extract.

4. A topical composition according to claim 1, wherein the extract of Cassia occidentalis is a pod extract.

5. A topical composition according to claim 1, wherein the extract of Cassia alata is a leave extract.

6. A topical composition according to claim 1, wherein the weight ratio of Coleus forskohlii extract:Cassia occidentalis and/or Cassia alata represents 3:1.

7. A topical composition according to claim 1, wherein: the Coleus forskohlii extract represents between 5.0 and 8.0% in weight of the composition; and the Cassia occidentalis and/or Cassia alata extract represents between 2.0 and 5.0% in weight of the composition.

8. A topical composition according to claim 1, wherein the penetration enhancer is diethylene glycol monoethyl ether and represents between 0.5 and 5.0% in weight of the composition, advantageously between 1.0 to 2.0%.

9. A topical composition according to claim 1, wherein the pH is ranging between 6.5 to 7.1.

10. A topical composition according to claim 1, wherein said composition is formulated as a leave-on product or a rinse-off product.

11. A topical composition according to claim 1, for use to promote the proliferation of melanocytes in the skin and/or in the hair follicle unit.

12. A topical composition according to claim 1, for use to promote the melanogenesis in the skin and/or in the hair follicle unit.

13. A topical composition according to claim 1, for use to treat canitie.

14. A topical composition according to claim 1, for use as a tanning agent.

15. A topical composition according to claim 1, for use to treat skin disease related to depigmentation in a human subject.

16. A topical composition for use according to claim 15, wherein skin disease related to depigmentation is selected from the group consisting of pytiriasis alba, pytiriasis versicolor, idiopathic guttate hypomelanosis, progressive macular hypomelanosis, post-inflammatory hypopigmentation and vitiligo.

17. A topical composition for use according to claim 15, wherein skin disease related to depigmentation is vitiligo.

Description

FIGURES

[0096] FIG. 1 depicts human melanocytes cultured during 96 hours with A/Control (NaCI 0.9% solution) B/Cassia occidentalis extract (5 g/ml) C/Coleus forskohlii extract (5 g/ml) and D/Coleus forskohlii extract (5 g/ml)+Cassia occidentalis extract (5 g/ml).

[0097] FIG. 2 depicts melanin contents of BC16 melanoma murine cells treated during 96 hours in presence of control (NaCI 0.9% solution), Coleus forskohlii extract (C. forskohlii; 5 g/ml); Cassia occidentalis extract (C. occidentalis; 5 g/ml); Cassia alata extract (C. alata; 5 g/ml); the combination of Coleus forskohlii extract (5 g/ml)+Cassia occidentalis extract (5 g/ml) or Cassia alata extract (5 g/ml)diethylene glycol monoethyl ether (DGEE; 1.5%).

[0098] FIG. 3 depicts the values of Microphthalmia-associated transcription factor (Mitf), Tyrosinase-related protein-1 (TRP-1), Tyrosinase-related protein-2 (TRP-2) and Tyrosinase (TYR) in A375 melanocytes after 96 hours in culture with control (NaCI 0.9%), Cassia occidentalis extract (5 g/ml), Coleus forskohlii extract (5 g/ml) and combination of Cassia occidentalis extract (5 g/ml) and Coleus forskohlii (5 g/ml).

[0099] FIG. 4 depicts the levels of Transforming Growth Factor- in skin biopsies of normal skin and lesional skin of vitiligo patients, and after treatment of the lesional melanocytes with Cassia occidentalis extract (5 g/ml), Coleus forskohlii extract (5 g/ml), and the combination of both Cassia occidentalis extract (5 g/ml) and Coleus forskohlii extract (5 g/ml).

[0100] FIG. 5 depicts the levels of Tumour Necrosis Factor-a in skin biopsies of normal skin and lesional skin of vitiligo patients, and after treatment of the lesional melanocytes with Cassia occidentalis extract (5 g/ml), Coleus forskohlii extract (5 g/ml), and the combination of both Cassia occidentalis extract (5 g/ml) and Coleus forskohlii extract (5 g/ml).

[0101] FIG. 6 depicts the levels of Interferon- in skin biopsies of normal skin and lesional skin of vitiligo patients, and after treatment of the lesional melanocytes with Cassia occidentalis extract (5 g/ml), Coleus forskohlii extract (5 g/ml), and the combination of both Cassia occidentalis extract (5 g/ml) and Coleus forskohlii extract (5 g/ml).

[0102] FIGS. 7 to 10 depict a subject with vitiligo before (FIGS. 7 and 9) and after (FIGS. 8 and 10) topical treatment with a composition of the present invention during 30 days.

DETAILED DESCRIPTION OF EXAMPLES

[0103] The application of the composition on the skin of patients afflicted by skin disease related to depigmentation, such as vitiligo, (for example once a day, or two daily applications) has shown response featuring the reduction of depigmentation and reduction in the progression of disease, and even partial repigmentation of white patches. In another aspect of the present invention, the use of the combination of these two vegetal active extracts under a cosmeceutically acceptable form stimulates the survival, proliferation and differentiation of melanocytes.

Example 1Effect of the Composition According to the Invention Comprising Coleus forskohlii Extract and Cassia occidentalis Extract or Cassia alata Extract on Melanocytes Proliferation

[0104] FIG. 1 depicts culture of human melanocytes treated with control, with each of the extracts, and with the combination of both extracts for 96 hours. Compared to control, Coleus forskohlii extract and Cassia occidentalis extract show increased melanocyte proliferation, and combination of Coleus forskohlii extract and Cassia occidentalis extract show increased melanocyte proliferation, more important than with each active ingredient taken separately.

Example 2Effect of the Composition According to the Invention Comprising Coleus forskohlii Extract and Cassia occidentalis Extract on Melanogenesis

[0105] FIG. 2 depicts melanogenesis in B16 melanoma murine melanocytes treated with control, with Coleus forskohlii extract, with Cassia occidentalis extract, with Cassia alata extract and with the combination of Coleus forskohlii extract+Cassia occidentalis extract or Coleus forskohlii extract+Cassia alata extract, diethylene glycol monoethyl ether. It is obvious that the combination of Coleus forskholii with Cassia occidentalis or Cassia alata stimulates melanogenesis in a more important manner than each extract taken separately, compared with control. Moreover, it is illustrated that both plant extracts (Coleus forskholii+Cassia occidentalis or Coleus forskholii+Cassia alata) act synergetically to increase the melanin synthesis. Besides, FIG. 2 demonstrates that the addition of diethylene glycol monoethyl ether (DGEE) improves the melanogenesis induced by the combination of Coleus forskholii+Cassia occidentalis or Coleus forskholii+Cassia alata.

Example 3Effect of the Composition According to the Invention Comprising Coleus forskohlii Extract and Cassia occidentalis Extract on the Expression of Molecular Factors Melanogenesis

[0106] Melanogenesis is a complex biological process where eumelanin and phaeomelanin derive from a common tyrosinase-dependent pathway with the same precursor, tyrosine. From dopaquinone, the eumelanin and phaeomelanin pathways diverge. Two enzymes crucial to eumelanogenesis are the tyrosinase-related proteins TRP1 (also known as GP75 or b-locus) and TRP2 (also known as dopachrome tautomerase, DCT).

[0107] Microphthalmia-associated transcription factor (Mitf) is involved in lineage-specific pathway regulation of melanocytes. All these factors are of utmost importance for the eumelanogenesis. As depicted in FIG. 3, the Applicant have discovered that Cassia occidentalis extract, as well as Coleus forskohlii extract, are able to increase the production of all four factors in A375 melanocytes. Unexpectedly, the combination of Cassia occidentalis extract with Coleus forskohlii extract is able to increase the production of all four aforementioned factors in a more important manner than each extract taken separately.

Example 4Effect of the Composition According to the Invention Comprising Coleus forskohlii Extract and Cassia occidentalis Extract on the Expression of TGF-

[0108] It is known that the levels of Transforming Growth Factor- are usually increased in the skin of patients with vitiligo. In non-segmental vitiligo, the body surface area of lesions is positively correlated with elevated TGF-1. Significant increase in the expression of TGF- was observed in active patients with vitiligo, whereas no change was observed in stable patients, suggesting that high levels of TGF- promote the progression of the disease. As shown in FIG. 4, the Applicant have been able to confirm these findings in skin biopsies of normal skin and lesional skin of patients with vitiligo. They have also observed that treatment of these skin explants with Cassia occidentalis extract was reducing the levels of TGF-, but at a lesser extent than Coleus forskohlii, and unexpectedly they discovered that the combination of Cassia occidentalis extract with Coleus forskohlii extract was able to reduce the skin levels of TGF- at a much higher extent that each of these two extracts taken separately.

Example 5Effect of the Composition According to the Invention Comprising Coleus forskohlii Extract and Cassia occidentalis Extract on the Expression of TNF-

[0109] Levels of TNF- were also shown to be increased in skin biopsies of vitiligo lesions compared to normal skin. Cases of refractory generalized vitiligo showed high tissue levels of TNF-. Considering these cases, patients with a strong TNF- staining were characterized by a higher vitiligo disease activity score than patients with a weak staining, which suggests a probable role of TNF- in the pathogenesis of vitiligo.

[0110] A study revealed significant increase in TNF- transcript and protein levels in vitiligo patients compared to controls. It is commonly thought that TNF- can inhibit melanogenesis by decreasing the intracellular levels of tyrosinase and tyrosinase-related protein 1, involved in both melanogenesis and prevention of melanocyte death. TNF--treated melanocytes show marked cellular shrinking and reduced melanin production in vitro, as well as downregulation of Mitf, a transcription factor essential in the regulation of melanocyte development, proliferation, death, and melanogenesis.

[0111] As depicted in FIG. 5, the Applicant have been able to confirm higher levels of TNF- in biopsies taken from lesional skin of vitiligo patients, compared to normal skin of the same patients. Further, treating these explants with Cassia occidentalis extract, these levels of TNF- were slightly decreased. The reduction was much more important when treated with Coleus forskohlii extract, and unexpectedly the Applicant discovered that treatment with the combination of Cassia occidentalis extract and Coleus forskohlii extract was leading to a decrease in TNF- much more substantial than the decrease observed with each of these extracts taken separately.

Example 6Effect of the Composition According to the Invention Comprising Coleus forskohlii Extract and Cassia occidentalis Extract on the Expression of IFN-

[0112] It is well recognized that increased expression of the cytokine interferon- (IFN-) plays a pivotal role in vitiligo-induced depigmentation. It was demonstrated that IFN- inhibits melanogenesis in primary cultured human melanocytes by altering melanogenic enzyme mRNA expression and, more importantly, that IFN- directly induces melanocyte apoptosis. It is suggested that IFN- inhibits both basal and -MSH-induced melanogenesis by inhibiting MITF expression and that IFN- plays a role in controlling inflammation- or UV-induced pigmentary changes. The Applicant have also demonstrated in skin explants from lesional skin of vitiligo patients increased levels of IFN- compared with normal skin of the same patients. As depicted in FIG. 6, treating these explants with Cassia occidentalis extract leaded to a decrease in the level of IFN- more or less similar to that produced by treatment with Coleus forskohlii extract. Unexpectedly the Applicant found that the association of both extracts leaded to a higher reduction of IFN- levels.

Example 7Effect of the Composition According to the Invention Comprising Coleus forskohlii Extract and Cassia occidentalis Extract to Treat Vitiligo

[0113] FIGS. 7 to 10 depict the results of increased melanogenesis and melanocyte proliferation four weeks after topical application once a day of the composition of the invention to two subjects with vitiligo.

The treatment of vitiligo with a composition comprising a combination of Coleus forskohlii extract, Cassia occidentalis extract and/or Cassia alata extract in a cosmeceutically acceptable vehicle containing also a penetration enhancer, and whose pH is maintained as close as possible to 6.8 was found to be successful.

Example 8Comparative Study of the Composition According to the Invention with Other Compositions

[0114] 8.1Composition According to the Invention

[0115] At ambient temperature, and under light stirring, in 300 g (30%) of distilled water, are added respectively: [0116] 3.5 g (0.35%) of a gelling agent featuring a polyvinyl carboxy polymer crosslinked with ethers of pentaerythritol, known under the commercial brand Carbomer 940. [0117] 80 g (8%) of an extract of Coleus forskohlii obtained by the extraction of dried roots of the plant by a blend of ethanol and propylene glycol. [0118] 50 g (5%) of an extract of Cassia occidentalis and/or Cassia alata obtained by the extraction of dried pods or leaves of these plants by a methanolic solution. [0119] 10 g (1%) of a liquid preservative agent named phenoxyethanol. [0120] 1 g (0.1%) of an ion chelator known as disodium EDTA. [0121] 15 g (1.5%) of a penetration enhancer known under the name of diethylene glycol monoethyl ether.

[0122] One stirs slowly at ambient temperature, until a homogenous mixture is obtained.

[0123] Separately, under rapid stirring (1000 rpm) and at 80 C. are mixed in this order: [0124] 10 g (1%) of an excipient made of cetyl alcohol. [0125] 90 g (9%) of another excipient made of propylene glycol dipelargonate. [0126] 80 g (8%) of another excipient made of decyl oleate. [0127] 80 g (8%) of another excipient made of isostearyl isosterate.

[0128] Under rapid stirring, a perfect homogenisation can be obtained.

[0129] Then, distilled water heated to 82 C. is added in the above mixture in a quantity of 280.5 g (28.05%) under rapid stirring.

[0130] When the emulsion is constituted, and temperature back to 30-35 C. is added the first phase containing the vegetal active ingredients and then are added 6 grams (0.6%) of a 50% solution of triethanolamine in order to adjust the pH.

[0131] Slow stirring is continued still reaching the temperature of 25 C.

[0132] This composition is an oil-in-water emulsion, the colour is beige, it is smooth, unctuous, semi-liquid and its pH is ranging from 6.5 to 7.1 Its odour is sui generi.

[0133] This preparation is stable over time and may be applied on the skin once or twice daily during an unlimited period of time. One can observe, sometimes as soon as ten days, more generally in the course of the month following its application, a limitation and even a stop in depigmentation and usually a visible repigmentation as seen in FIGS. 7 to 10.

[0134] Further applying this emulsion prevents further depigmentation.

[0135] 8.2Composition Comprising Solely Coleus forskohlii Extract

[0136] Example 1 is repeated incorporating in the emulsion solely Coleus forskohlii extract. The effect is more limited and appears after a longer time.

[0137] 8.3Composition Comprising Solely Cassia occidentalis Extract

[0138] Example 1 is repeated incorporating in the emulsion solely Cassia occidentalis extract. The effect is very limited and appears after a longer time.

[0139] 8.4Composition Comprising Coleus forskohlii Extract and Cassia occidentalis Extract

[0140] Example 1 is repeated without incorporation of diethylene glycol monoethyl ether. The emulsion obtained has a pretty similar appearance and the same physical properties than in example 1. However, a limitation of depigmentation is observed in a slower and more sporadic manner than in example 1.

[0141] 8.5Composition According to the Invention with an Acidic pH

[0142] Example 1 is repeated without adjusting the pH between 6.5 to 7.1, but with a pH value more acidic, i.e. between 5.5 and 6.5. The emulsion obtained has a pretty similar appearance and the same physical properties than in example 1 (except for its pH).

[0143] However, a limitation of depigmentation is observed in a slower and more sporadic manner than in example 1.

[0144] 8.6Composition without any Plant Extracts According to the Invention

[0145] Example 1 is repeated replacing the vegetal extracts of Coleus forskohlii and Cassia occidentalis by other vegetal extracts. No result is observed.

[0146] These examples perfectly demonstrate the activity related to the synergistic effect resulting from the combination in the composition of Coleus forskohlii extract with Cassia occidentalis and/or Cassia alata extract with diethylene glycol monoethyl ether to enhance the penetration, and with a pH as close as possible to 6.8 on the limitation of depigmentation and the reduction of the inflammatory front in the course of the process of development of vitiligo and its further repigmentation.