COSMETIC USE OF AN AROMATIC RAW MATERIAL EXTRACT FOR PROTECTING SKIN FROM BLUE LIGHT

Abstract

Methods for protecting the skin from blue light comprising administering to a subject in need thereof at least one extract of aromatic raw material selected from the group consisting of a plant extract of rosemary containing at least 1% of rosmarinic acid, a black pepper absolute, a gentian absolute, a myrrh resinoid, an essential oil of mandarin petitgrain, a ginger absolute, and a tea resinoid.

Claims

1. A method for protecting the skin from blue light comprising administering to a subject in need thereof at least one extract of aromatic raw material selected from the group consisting of a plant extract of rosemary containing at least 1% of rosmarinic acid, a black pepper absolute, a gentian absolute, a myrrh resinoid, an essential oil of mandarin petitgrain, a ginger absolute, and a tea resinoid.

2. The method according to claim 1, wherein the plant extract of rosemary contains at least 5% rosmarinic acid.

3. The method according to claim 1, wherein the plant extract of rosemary is obtained from spent grains of Rosmarinus officinalis needles.

4. The method according to claim 1, wherein the black pepper absolute is obtained from Piper nigra berries.

5. The method according to claim 1, wherein the gentian absolute is obtained from roots of Gentiana lutea.

6. The method according to claim 1, wherein the resinoid myrrh is obtained from Commiphora myrrha resin.

7. The method according to claim 1, wherein the essential oil of mandarin petitgrain is obtained by steam distillation of leaves of Citrus reticulata.

8. The method according to claim 1, wherein the ginger absolute is obtained from rhizomes of Zingiber officinale.

9. The method according to claim 1, wherein the ginger absolute contains more than 40% dry matter.

10. The method according to claim 1, wherein the tea resinoid is obtained from leaves of Camelia sinensis.

11. The method according to claim 1, wherein the at least one extract of aromatic raw material comprises the plant extract of rosemary, the black pepper absolute, and the gentian absolute.

12. The method according to claim 1, wherein the at least one extract of aromatic raw material comprises the plant extract of rosemary, the black pepper absolute, the gentian absolute, and the myrrh resinoid.

13. The method according to claim 1, wherein the at least one extract of aromatic raw material comprises the plant extract of rosemary, the black pepper absolute, the gentian absolute, the myrrh resinoid, the essential oil of mandarin petitgrain, and the ginger absolute.

14. The method according to claim 1, wherein the at least one extract of aromatic raw material comprises the plant extract of rosemary, the black pepper absolute, the gentian absolute, the myrrh resinoid, the essential oil of mandarin petitgrain, the ginger absolute, and the tea resinoid.

15. A cosmetic composition comprising at least three extracts selected from the group consisting of a plant extract of rosemary containing at least 1% of rosmarinic acid, a black pepper absolute, a gentian absolute, a myrrh resinoid, an essential oil of mandarin petitgrain, a ginger absolute, and a tea resinoid.

16. The cosmetic composition according to claim 15, wherein the at least three extracts comprise the plant extract of rosemary, the black pepper absolute, and the gentian absolute.

17. The cosmetic composition according to claim 16, wherein the at least three extracts further comprise the myrrh resinoid, the essential oil of mandarin petitgrain, the ginger absolute, and the tea resinoid.

18. The cosmetic composition according to claim 15, wherein the plant extract of rosemary is obtained from spent grains of Rosmarinus officinalis needles.

19. The cosmetic composition according to claim 15, wherein the black pepper absolute is obtained from Piper nigra berries.

20. The cosmetic composition according to claim 15, wherein the gentian absolute is obtained from roots of Gentiana lutea.

21. The cosmetic composition according to claim 15, wherein the resinoid myrrh is obtained from Commiphora myrrha resin.

22. The cosmetic composition according to claim 15, wherein the essential oil of mandarin petitgrain is obtained by steam distillation of leaves of Citrus reticulata.

23. The cosmetic composition according to claim 15, wherein the ginger absolute is obtained from rhizomes of Zingiber officinale.

24. The cosmetic composition according to claim 15, wherein the tea resinoid is obtained from leaves of Camelia sinensis.

Description

EXAMPLE 1—COMPOSITIONS OF THE INVENTION

Preparation

[0307] A composition comprising: [0308] a resinoid obtained from leaves of Camelia sinensis—commercial reference R30632 (from Robertet); [0309] a plant extract obtained from spent grains of Rosmarinus officinalis needles—commercial reference R30565 (from Robertet); [0310] an absolute obtained from Piper nigra berries—commercial reference R21289 (from Robertet); [0311] an absolute obtained from the roots of Gentiana lutea—commercial reference R20363 (from Robertet); [0312] an absolute obtained from rhizomes of Zingiber officinale—commercial reference R20407 (from Robertet); [0313] a resinoid obtained from Commiphora myrrha resin—commercial reference R10896 (from Robertet); and [0314] an essential oil obtained by steam distillation of the leaves, branches and green fruits of Citrus reticulata—commercial reference R21500 (from Robertet).

[0315] To do this, the different ingredients are weighed and then mixed together. A neutral solvent (i.e. 96.2% ethyl alcohol) is added to facilitate the dissolution of the ingredients, some of which are more or less pasty and/or solid.

[0316] The composition reported in the following Table 1 is obtained.

TABLE-US-00001 TABLE 1 Composition of the invention Ingredient % (p/p) R30632 10% R30565 19% R21289 10% R20363  1% R20407 10% R10896 10% R21500 10% Ethyl alcohol 96.2% 30%

EXAMPLE 2—IN VIVO EFFICACY OF THE COMPOSITIONS OF THE INVENTION

2.1—Experimental Protocol

[0317] Primary human dermal fibroblasts, from a 57-year-old female donor, are thawed and amplified in flasks for a few days.

[0318] 24 hours before the start of the test, each flask is treated as follows: [0319] Flask n° 1: culture medium; [0320] Flask n° 2: composition of example 1 at 30 ppm; [0321] Flask n° 3: composition of example 1 at 20 ppm; [0322] Flask n° 4: composition of example 1 at 10 ppm; [0323] Flask n° 6: composition of example 1 at 2.5 ppm.

[0324] Day 1

[0325] The cells are incubated with the «Mitotracker Green» dye for 15 minutes.

[0326] Cells are washed with PBS, detached and seeded in a CYTOOplate with extra large Y microstructures at 3,000 cells/well in 10% serum.

[0327] 1 h30 later, once the cells are attached and diffused on the microstructure, the medium is changed to medium containing less serum plus the active ingredient at 5 concentrations, incubated for 2 h at 37 degrees with 5% CO.sub.2.

[0328] After 2 hours of treatment, the cells are irradiated with LEDs emitting at 447 nm for 1 hour corresponding to 20 mJ/cm2.

[0329] Hoechst is added for 15 min in each well to stain the nucleus

[0330] The medium is changed to wash off the nuclear stain and the cells are incubated in medium plus active ingredient.

[0331] Live imaging was performed on the LEICA microscope under a controlled environment at objective 40, with 49 fields/wells.

[0332] At the end of the imaging, the cells are fixed and the actin filaments are labeled with Phalloidin 555. The images were acquired on the Operetta HCS platform from Perkin Elmer.

Blue Light Stress

[0333] A device developed by Genel is used to irradiate cells with 447 nm light. The device is made of LEDs and delivers a dose of 20 J/cm2.

[0334] Radiant light is the power of light received on a surface per unit area [W/cm2]=Dose [J/cm.sup.2]/time in seconds.

[0335] The radiant light emitted by a screen is about 18 to 40 μW/cm2 at a distance of 10 centimeters. The average screen usage time is 5 hours per day, i.e. 18,000 seconds per day, namely a daily dose emitted by a screen at 10 cm=light emitted*time=40 μW/cm2*18000 s/day=0.72 J/cm2/day.

[0336] By using the device at 20 J/cm.sup.2 for 1 hour, it is simulated that the dose of 1 month (28 days) of screen at a distance of 10 cm.

Mitochondrial Characterization by Image Analysis

[0337] A very large reading panel was developed with the aim of collecting as much information as possible about mitochondrial filament size and density to detect fragmentation and elongation phenotypes.

[0338] The mitochondrial network can be divided into several groups of filaments which are linked together: the basal unit is called «Tree». The number of Trees per array, as well as their total length (Average Total Tree Length) is an average across all single cells detected in each well.

[0339] Each Tree is divided into branches that define each other by either a junction or an endpoint. These branches are characterized by measuring their maximum size in the total network of a single cell.

2.2—Results

Observations

[0340] Mitochondrial fluorescence images were acquired at the objective 40.

[0341] The mitochondrial network of untreated unstressed cells is nicely elongated/with nice extensions.

[0342] Cells treated with the composition of example 1 but not stressed by blue light show a well extended mitochondrial network similar to the control. (Negative control)

[0343] Blue light stress induces dramatic mitochondrial fragmentation. The number of «subunits» is increased by 3 and their length is reduced almost 4 times.

[0344] The same trend is detected on the measurement of ramifications with an average length decreased by 1.7 times.

[0345] Cells were treated in the flask with the composition of Example 1 for 24 h prior to plating as well as during blue light stress.

[0346] Treatment with the composition of Example 1 partially protects the cellular mitochondrial network from fragmentation by blue light.

Statistics

[0347] Throughout the study, statistical tests were carried out according to:

[0348] 1) An unpaired T-test followed by a Welch correction to compare:

[0349] The «untreated unstressed» condition with the «untreated stressful light» condition

[0350] 2) An Anova to compare: [0351] the «untreated, unstressed» condition with the «composition of example 1» condition; [0352] the «untreated light stressed» condition with the «composition of example 1 in the ppm light stress» conditions;

[0353] with *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

[0354] The results obtained are reported in Tables 2 to 9 below.

TABLE-US-00002 TABLE 2 NB Tree/Total (without stress) Doses of composition Untreated sample according to example 1 (in ppm) (without stress) 2.5 5 10 20 30 0.135 0.140 0.088 0.093 0.108 0.114 0.099 0.129 0.107 0.092 0.107 0.113 0.119 0.095 0.108 0.103 0.141 0.134 0.112 0.108 0.093 0.102 0.146 0.194 0.142 0.132 0.131 0.108 0.162 0.158 Average 0.121 0.121 0.105 0.099 0.133 0.143

TABLE-US-00003 TABLE 3 NB Tree/Total (with stress) Doses of composition Untreated sample Untreated sample according to example 1 (in ppm) (without stress) (with stress) 2.5 5 10 20 30 0.135 0.436 0.326 0.290 0.335 0.407 0.217 0.099 0.392 0.334 0.297 0.301 0.349 0.235 0.119 0.311 0.274 0.225 0.227 0.227 0.195 0.112 0.476 0.297 0.253 0.322 0.305 0.249 0.142 0.409 0.310 0.287 0.309 0.342 0.368 Average 0.121 0.405 0.308 0.270 0.299 0.326 0.253 % — — 34.1 47.4 37.3 27.8 53.6 protected

TABLE-US-00004 TABLE 4 Average Tree Length (without stress) Doses of composition Untreated sample according to example 1 (in ppm) (without stress) 2.5 5 10 20 30 7.469 9.739 12.42 11.187 10.506 9.868 10.905 10.234 9.808 10.653 9.594 9.138 9.022 11.688 10.61 10.396 7.749 8.416 9.347 10.204 11.62 11.911 6.918 5.972 7.565 8.252 8.399 10.195 6.475 6.739 Average 8.862 10.023 10.571 10.868 8.248 8.027

TABLE-US-00005 TABLE 5 Average length of Tree (with stress) Doses of composition according to Untreated sample Untreated sample example 1 (in ppm) (without stress) (with stress) 2.5 5 10 20 30 7.469 2.166 3.14 3.437 2.996 2.556 4.977 10.905 2.442 2.952 3.431 3.304 2.833 4.27 9.022 3.313 3.496 4.486 4.087 4.325 5.255 9.347 2.141 3.281 3.861 3.067 3.202 4.116 7.565 2.222 3.32 3.36 3.255 2.834 2.8 Average 8.862 2.457 3.238 3.715 3.342 3.150 4.284 % — — 12.2 19.6 13.8 10.8 28.5 protected

TABLE-US-00006 TABLE 6 NB branches/Total (without stress) Doses of composition Untreated sample according to example 1 (in ppm) (without stress) 2.5 5 10 20 30 0.448 0.473 0.467 0.467 0.495 0.485 0.428 0.44 0.433 0.48 0.458 0.469 0.453 0.45 0.448 0.45 0.488 0.478 0.439 0.441 0.461 0.461 0.482 0.555 0.453 0.443 0.46 0.47 0.486 0.491 Average 0.444 0.449 0.454 0.466 0.482 0.496

TABLE-US-00007 TABLE 7 NB branches/Total (with stress) Doses of composition according to Untreated sample Untreated sample example 1 (in ppm) (without stress) (with stress) 2.5 5 10 20 30 0.448 0.764 0.642 0.585 0.673 0.742 0.59 0.428 0.747 0.664 0.609 0.631 0.701 0.616 0.453 0.677 0.59 0.531 0.566 0.609 0.568 0.439 0.752 0.614 0.551 0.627 0.672 0.617 0.453 0.749 0.578 0.58 0.586 0.672 0.673 Average 0.444 0.738 0.618 0.571 0.617 0.679 0.613 % — — 40.9 56.7 41.3 20.0 42.6 protected

TABLE-US-00008 TABLE 8 Average branch length (without stress) Doses of composition Untreated sample according to example 1 (in ppm) (without stress) 2.5 5 10 20 30 2.22 2.288 1.965 1.961 1.836 1.961 2.135 2.232 2.192 1.848 2.002 1.949 2.061 2.06 2.061 2.034 1.846 1.998 2.087 2.214 2.028 1.933 1.972 1.688 2.137 2.147 2.198 1.9 1.941 1.883 Average 2.128 2.188 2.089 1.935 1.919 1.896

TABLE-US-00009 TABLE 9 Average branch length (with stress) Doses of composition according to Untreated sample Untreated sample example 1 (in ppm) (without stress) (with stress) 2.5 5 10 20 30 2.22 1.157 1.44 1.65 1.333 1.151 1.575 2.135 1.182 1.333 1.576 1.41 1.217 1.432 2.061 1.348 1.586 1.767 1.639 1.487 1.526 2.087 1.18 1.455 1.706 1.483 1.294 1.391 2.137 1.156 1.642 1.639 1.545 1.258 1.412 Average 2.128 1.205 1.491 1.668 1.482 1.281 1.467 % 31.0 50.1 30.0 8.3 28.4 protected

Conclusion

[0355] Blue light stress induces strong fragmentation of the mitochondrial network in fibroblasts, increasing the number of mitochondrial subunits. Treatment with the composition of example 1 protects the cells from stress by blue light with a maximum effect at 5 ppm.

EXAMPLE 3—IN VIVO EFFICACY OF THE COMPOSITIONS OF THE INVENTION

3.1—Experimental Protocol

[0356] At the end of the experiment detailed in Example 2, the plates are fixed in a formaldehyde solution and the cells are stained with Phalloidin. Images are acquired at *20 with the Operetta HCS instrument (Perkin Elmer) in three channels: microstructure, nucleus and F-actin.

[0357] A special analysis of the images was carried out with the aim of detecting single cells in the microstructure and measuring their area. The correctly plated cells counted have an area about 1800 μm.sup.2.

3.2—Results

[0358] The obtained results are reported in Tables 10 to 13 below.

TABLE-US-00010 TABLE 10 Cell surface (without stress) Doses of composition Untreated sample according to example 1 (in ppm) (without stress) 2.5 5 10 20 30 2062 2120 2204 2371 2242 2177 2071 2071 2255 2417 2226 2292 2019 2120 2244 2338 2429 2341 2161 2155 2155 2256 2286 2119 2007 2066 2140 2119 2147 2324 Average 2064 2106 2200 2300 2266 2250

TABLE-US-00011 TABLE 11 Cell surface (with stress) Doses of composition according to Untreated sample Untreated sample example 1 (in ppm) (without stress) (with stress) 2.5 5 10 20 30 2062 1779 2022 2083 2113 1978 2062 2071 1906 2069 2130 2026 2016 2071 2019 2033 2078 2226 2283 2231 2019 2161 1913 2107 2122 2084 2009 2161 2007 1759 1936 2076 1918 1821 2007 Average 2064 1878 2042 2127 2085 2011 2064 % — — 88.5 134.2 111.2 71.5 3.5 protected

TABLE-US-00012 TABLE 12 % Cell Diffusion > 1800 (without stress) Doses of composition Untreated sample according to example 1 (in ppm) (without stress) 2.5 5 10 20 30 63.64 72.90 81.69 90.14 81.10 80.86 71.74 70.67 77.12 93.13 81.25 82.93 64.03 71.43 80.81 84.91 91.94 87.13 72.22 72.55 77.27 81.33 86.71 76.87 66.46 71.35 70.39 77.54 75.28 76.70 Average 67.62 71.78 77.46 85.41 83.25 80.90

TABLE-US-00013 TABLE 13 % Cell Diffusion > 1800 (with stress) Doses of composition according to Untreated sample Untreated sample example 1 (in ppm) (without stress) (with stress) 2.5 5 10 20 30 63.64 41.73 66.43 71.43 71.59 67.38 57.75 71.74 50.00 63.97 73.10 66.85 66.99 54.93 64.03 58.23 70.70 83.12 88.36 82.04 68.06 72.22 50.00 69.14 72.97 75.31 65.73 64.61 66.46 39.41 58.38 69.94 66.44 55.21 43.52 Average 67.62 47.87 65.73 74.11 73.71 67.47 57.78 % — — 90.4 132.9 130.9 99.3 50.2 protected

[0359] In the untreated condition, the cell surface is around 2063 μm.sup.2 and 67% of the cells alone are nicely dispersed on the microstructure, with actin filaments on the corners.

[0360] The addition of the composition according to Example 1 without stress induces an increase in the cell surface as well as cell diffusion in a dose/response pathway of up to 10 ppm. The slight toxicity detected on the mitochondrial network has no impact on cell diffusion.

[0361] The stressful blue light negatively impacts cell diffusion capacity, reducing the number of diffused beautiful cells by 48%. The treatment with the composition according to Example 1 completely cancels the effects of blue light on cell diffusion at 5 and 10 ppm, with a viability percentage of 132% at 5 ppm.

3.3—Conclusion

[0362] The F-actin cytoskeleton is also impacted by stressful blue light, reducing the ability of cells to diffuse. The composition according to Example 1 also protects the fibroblasts from the stress induced by blue light on their diffusion capacities. Maximum effect was again observed at 5 ppm.

EXAMPLE 4—COMPARATIVE EXAMPLE

[0363] We are trying to determine the absorption spectra of the following extracts: [0364] a plant extract obtained from spent grains of Rosmarinus officinalis needles—commercial reference R30565 (from Robertet)—plant extract of rosemary; [0365] an essential oil obtained from the branches and leaves of Rosmarinus officinalis—commercial reference R21614 (from Robertet)—rosemary essential oil; [0366] an absolute obtained from Zingiber officinale rhizomes—commercial reference R20407 (from Robertet)—ginger absolute; and [0367] an essential oil obtained from the roots of Zingiber officinale—commercial reference R20779 (at Robertet)—essential oil of ginger.

[0368] To do this, each sample is diluted to 1000 ppm in ethanol. Each dilution is then transferred to a well of a 96-well microplate. An absorption spectrum is then obtained using a TECAN Infinite 2000 pro spectrophotometer. The OD (Optical Densities) are measured in the wavelengths of the visible spectrum (400 nm-800 nm).

[0369] The absorption spectra obtained for plant extract of rosemary, rosemary essential oil, ginger absolute and ginger essential oil are respectively reported in FIGS. 1 to 4.

[0370] It appears that the plant extract of rosemary has an absorption between 400 and 500 nm, which confirms that this extract protects the skin against blue light. On the contrary, rosemary essential oil shows no absorption between 400 and 500 nm, and therefore cannot be used for protecting the skin from blue light.

[0371] Similarly, it appears that ginger absolute has an absorption between 400 and 500 nm, which confirms that this extract protects the skin against blue light. On the contrary, ginger essential oil shows no absorption between 400 and 500 nm, and therefore cannot be used for protecting the skin from blue light.