USE OF MAGNOLIA, JUNIPER OR VIOLET ABSOLUTE IN COSMETICS FOR DEPIGMENTING THE SKIN

Abstract

The cosmetic use of one or more absolute(s) obtained from magnolia, juniper or violet for depigmenting the skin.

Claims

1. A cosmetic method comprising applying one or several absolute(s) obtained from magnolia, juniper or violet to depigment the skin.

2. The method according to claim 1, in that wherein the absolute obtained from magnolia is obtained from the magnolia bark.

3. The method according to claim 2, where the absolute obtained from magnolia is obtained from the Magnolia officinalis bark.

4. The method according to claim 1, wherein the absolute obtained from juniper is obtained from the juniper berry.

5. The method according to claim 4, wherein the absolute obtained from juniper is obtained from the Juniperus communis berries.

6. The method according to claim 1, wherein the absolute obtained from violet is obtained from the violet leaves.

7. The method according to claim 6, wherein the absolute obtained from violet is obtained from the Viola odorata leaves.

8. A cosmetic method comprising applying an absolute obtained from magnolia as defined in claim 1 combined with an absolute obtained from juniper to depigment the skin.

9. A cosmetic method comprising applying an absolute obtained from magnolia as defined in claim 1 combined with an absolute obtained from violet to depigment the skin.

10. A cosmetic method comprising applying an absolute obtained from juniper as defined in claim 1 combined with an absolute obtained from violet to depigment the skin.

11. A cosmetic method comprising applying an absolute obtained from magnolia as defined in claim 1 combined with an absolute obtained from juniper and with an absolute obtained from violet to depigment the skin.

Description

EXAMPLE 1EFFECTIVENESS IN VIVO OF THE COMPOSITIONS OF THE INVENTION

[0058] Preparation of the Magnolia Absolutes

[0059] In a 20 liter reactor, 3 kg of crushed Magnolia barks are extracted with 5 volumes of hexane at ambient temperature and under mechanical stirring for 1 h30. The whole is left to settle for 30 minutes and then filtered on a pleated filter. The exhausted barks are extracted again with 5 volumes of hexane under the same conditions as previously described.

[0060] The two extraction filtrates are combined together and vacuum concentrated in the rotary evaporator (40 C. under 30 mbar).

[0061] 186.3 g of a green-brown viscous liquid (concrete) are obtained, that is to say a 6.2% yield.

[0062] In a 4 liter reactor provided with a mechanical stirring, the concrete thus obtained (186.3 g) is dissolved in 6 volumes of denatured alcohol and stirred for 1 h at 45 C.

[0063] Then, the whole is left to ice over in the freezer for 12 hours.

[0064] The solution is filtered on a sinter provided with a Celite bed. Thus, a first filtrate (S1) is collected.

[0065] The residual waxes retained on the sinter are washed again with 2 volumes of cold denatured alcohol. Thus, a second filtrate (S2) is collected.

[0066] The two filtrates are vacuum concentrated separately in the rotary evaporator (50 C. under 30 mbar). A solubility test is performed on the two concentrated filtrates individually to check up that they have no insoluble matter when they are put in solution at 10% in the alcohol.

[0067] The two concentrated filtrates are then mixed and 114.3 g of an orange-brown liquid is obtained, that is to say a 61.3% yield (Magnolia absolute).

[0068] Preparation of the Juniper and Violet Absolutes

[0069] The method described hereinbefore has also allowed obtaining the following absolutes: [0070] Juniper absolute corresponding to the absolute obtained from Juniperus communis berries; and [0071] Violet absolute obtained from Viola odorata leaves.

EXPERIMENTAL PROTOCOL

[0072] The following protocol has allowed determining the half-maximal inhibitory concentration (IC50) of the different absolutes, alone or in combination. The IC.sub.50 is a measurement of the effectiveness of a given compound to inhibit a specific biological or biochemical function. In this instance, IC50 corresponds to the concentration at which the ratio for the maximum rate of transformation of the substrate into product for the condition on the Vmax of the transformation for the control is 50%.

[0073] In vivo model tests on melanin-producing cells, the murine melanocytes B16-F10, have been necessary. The decrease in the cellular tyrosinase activity, associated with a reduction of the intracellular melanin content, while preserving the survival of the cells by non-cytotoxic concentrations, have then been investigated. For this purpose, the B16-F10 cells have been seeded into 24-well plates, and incubated for 24 h at 37 C. with several concentrations of different selected raw materials. The kojic acid has also been used as reference product. Afterwards, the cells are lysed, and centrifuged.

[0074] The supernatant contains tyrosinase that is deposited in a 96-well plate and to which L-DOPA substrate is added, and then the absorbance at 490 nm is read every 10 min for 1 hour. The maximum rate of the condition/maximum rate of the negative control ratio, gives the inhibition rate of the condition and allows estimating the IC50 of each raw material.

[0075] In turn, the cell pellet is dissolved by a NaOH 1N solution and incubated for 1 hour at 70 C. before being deposited in a 96-well plate put in the spectrophotometer at 405 nm. Afterwards, a comparison with the absorbance of a standard melanin calibrated range allows determining the intracellular melanin content of each condition and checking on the inhibitor effect of the melanogenesis by the different raw materials.

[0076] A cell viability test in tetrazolium MTT salt (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) is performed. For this purpose, the cells are incubated at 37 C. for 3 h in a medium containing 1 mg/ml of MTT, before solubilizing the formed formazan crystals by the DMSO and reading the absorbance at 570 nm. The cell survival rate is expressed in percentage with respect to the negative controls.

[0077] Results

[0078] The obtained results are reported in the following Tables 1 and 2, as well as in FIG. 1 representing photographs of control murine cells B16-F10 (A) and incubated with the mixture Magnolia Absolute/Juniper Absolute/Violet Absolute (1/1/1) (B).

TABLE-US-00001 TABLE 1 Tests on B16 melanocytes, with the assessment of the activity of the intracellular tyrosinase Effective Theoretical Ingredient IC50 (ppm) IC50 Magnolia Absolute 18 Juniper Absolute 9 Violet Absolute 17 Magnolia Absolute/Juniper Absolute (1/1) 7 12 Magnolia Absolute/Violet Absolute (1/1) 10 17 Violet Absolute/Juniper Absolute (1/1) 4 13 Magnolia Absolute/Juniper Absolute/ 4.2 13 Violet Absolute (1/1/1) Kojic acid 225

TABLE-US-00002 TABLE 2 Tests on B16 melanocytes, with dosing of the intracellular melanin. Effective Theoretical Ingredient IC50 (ppm) IC50 Magnolia Absolute 18.5 Juniper Absolute 111 Violet Absolute 54 Magnolia Absolute/Juniper Absolute (1/1) 20 32 Magnolia Absolute/Violet Absolute (1/1) 18 28 Violet Absolute/Juniper Absolute (1/1) 52 73 Magnolia Absolute/Juniper Absolute/ 21 37 Violet Absolute (1/1/1) Kojic acid 214

CONCLUSION

[0079] The obtained results confirm the inhibitory activity of tyrosinase and of the synthesis of the intracellular melanin of each of the tested absolutes, and therefore their cosmetic effectiveness in depigmenting the skin.

[0080] Furthermore, these results highlight the existence of an unexpected synergy of action between the different absolutes. This synergy is observed for the combinations of two or three absolutes.