COSMETIC COMPOSITIONS IN PARTICULAR WITH ANTI-AGING EFFECT, COMPRISING A GREEN EXTRACT OF AFRAMOMUM ANGUSTIFOLIUM OR LONGOZA PLANT

Abstract

The present invention relates to a cosmetic composition.

This composition is characterized in that it comprises, as active ingredient, a green extract of seeds of the Aframomum angustifolium or longoza plant, obtained by extraction with a polar solvent of polyol, in particular a glycol, optionally in a cosmetically acceptable excipient.

This cosmetic composition has a good anti-aging effect.

Claims

1-10. (canceled)

11. A cosmetic care process for performing an anti-aging effect to combat the effects of aging resulting from mitochondrial dysfunction, comprising at least one topical application to the skin of a person in need thereof of a cosmetically effective amount for performing an anti-aging effect to combat the effects of aging resulting from mitochondrial dysfunction, of an extract, called green extract, of seeds of the Aframomum angustifolium or longoza plant made with an ecologically friendly and cosmetically acceptable polar solvent consisting essentially of a polyol, in a cosmetically acceptable excipient.

12. The process as claimed in claim 11, wherein the at least one topical application of the seed extract of Aframomum angustifolium or longoza plant is carried out on the areas of the skin in need of a cosmetic care selected from the group consisting of an anti-aging care, of an anti-wrinkle care to reduce, eliminate or slow down the appearance of wrinkles, of an action preserving or restoring the structure of the skin, of a protective, repairing or restoring effect on the elasticity or firmness of the skin.

13. The process as claimed in claim 11, wherein the polyol consists essentially of a glycol.

14. The process as claimed in claim 11, wherein the extract from the seeds of the plant is obtained by an extraction process comprising at least one step of extracting the ground seeds with the polar solvent of polyol.

15. The process as claimed in claim 11, wherein the extract from the seeds of the plant is dissolved again in an environmentally friendly and cosmetically acceptable solvent comprising, or consisting essentially of a polyol, or an alcohol or a hydroalcoholic mixture.

16. The process as claimed in claim 11, wherein the polyol has the general chemical formula HO—R—OH, wherein R represents a hydrocarbon radical —(CH.sub.2).sub.n with n having 2 to 16 carbon atoms.

17. The process as claimed in claim 16, wherein the polyol has the general chemical formula HO—R—OH, wherein R represents a hydrocarbon radical —(CH.sub.2).sub.n with n having 3 to 8 carbon atoms.

18. The process as claimed in claim 11, wherein the polyol comprises a glycol selected from the group consisting of propylene glycol, butylene glycol or 1,3-butylene glycol, pentylene glycol, and mixtures thereof.

19. The process as claimed in claim 11, wherein said extract of longoza seeds is present at a concentration, expressed as dry extract, of between 0.001% and 5%, by weight based on the total weight of the composition.

20. The process as claimed in claim 11, wherein said extract of longoza seeds is present at a concentration, expressed as dry extract, of between 0.01% and 1%, by weight based on the total weight of the composition.

21. A cosmetic care process for performing an anti-aging effect to combat the effects of aging resulting from mitochondrial dysfunction, comprising at least one topical application to the skin of a person in need thereof of a cosmetically effective amount for performing an anti-aging effect to combat the effects of aging resulting from mitochondrial dysfunction, of an extract, called green extract, of seeds of the Aframomum angustifolium or longoza plant made with an ecologically friendly and cosmetically acceptable polar solvent consisting essentially of butylene glycol, in a cosmetically acceptable excipient.

22. The process as claimed in claim 21, wherein said extract of longoza seeds is present at a concentration, expressed as dry extract, of between 0.001% and 5%, by weight based on the total weight of the composition.

23. The process as claimed in claim 21, wherein said extract of longoza seeds is present at a concentration, expressed as dry extract, of between 0.01% and 1%, by weight based on the total weight of the composition.

24. The process as claimed in claim 21, wherein the extract from the seeds of the plant is dissolved again in an environmentally friendly and cosmetically acceptable solvent comprising, or consisting essentially of a polyol, or an alcohol or a hydroalcoholic mixture.

25. The process as claimed in claim 24, wherein the polyol has the general chemical formula HO—R—OH, wherein R represents a hydrocarbon radical —(CH.sub.2).sub.n with n having 2 to 16 carbon atoms.

26. The process as claimed in claim 24, wherein the polyol has the general chemical formula HO—R—OH, wherein R represents a hydrocarbon radical —(CH.sub.2).sub.n with n having 3 to 8 carbon atoms.

27. The process as claimed in claim 24, wherein the polyol comprises a glycol selected from the group consisting of propylene glycol, butylene glycol or 1,3-butylene glycol, pentylene glycol, and mixtures thereof.

Description

EXAMPLE 1 ACCORDING TO THE INVENTION

[0048] Example of Laboratory Preparation of a Green Extract of Seeds of the Aframomum angustifolium or Longoza Plant, with a Glycol, for Example Butylene Glycol, According to the Invention.

[0049] From 200 g of longoza plant seeds from various commercial lots, the following extraction procedure is carried out:

[0050] a) first, the commercially available dried seeds are ground to an average powder size of, for example, 0.01 mm to 1 mm.

[0051] b) then, the powder is macerated overnight for about 12 hours in 350 mL of butylene glycol, or a seed/solvent ratio (w/v) of 1/1.75 at static room temperature.

[0052] c) after maceration, a first filtration is carried out, for example on a filter with a porosity of 20-25 micrometers, to obtain a first filtrate by obtaining an opaque orange-red solution with a dry residue around 20 g/L or maceration juice, and a cake of ground seeds, then

[0053] d) an extraction is carried out on the cake with a further 350 mL of butylene glycol, at a mild temperature, for example around 55° C. for a period of several hours, for example around 3 hours, and then

[0054] e) a second filtration is carried out, for example on a filter with a porosity of 20-25 micrometers, to obtain a second filtrate, obtaining an opaque orange solution with a dry residue of around 5 g/L.

[0055] f) According to a particular alternative, the two filtrates or extracts obtained are combined and may be re-filtered through a sterilizing filter, for example of 0.5 to 0.1 micrometers. The filtrate is a clear orangish-orange solution on which the active ingredient assay and thin-layer chromatography and microbiology can be carried out.

[0056] g) The filtrate or extract solution obtained may be adjusted with butylene glycol or other biocompatible and cosmetically acceptable solvent to give a dry matter content of 10 g/L or about 1% by weight of dry matter.

[0057] This butylene glycol extract constitutes in itself, according to a particular embodiment, the green extract of longoza or Aframomum angustifolium seeds according to the invention which can be used as a cosmetic agent or as an active ingredient for the manufacture of a cosmetic composition as shown in the following examples (1 g extract). This butylene glycol solution contains 1% by weight of dry extract.

[0058] h) for the incorporation of this dry extract into a cosmetic composition according to the invention, it is also possible to prepare a fresh solution thereof at the time of manufacturing said composition, in a cosmetically acceptable solvent, in particular in water, alcohol, 1,3-butylene glycol or a mixture of these solvents. This embodiment thus makes it possible to prepare solutions of dry green extract of the Aframomum angustifolium or longoza plant at the desired concentration.

[0059] i) finally, the various solutions of the extract described above may be packaged, pending their use in the manufacture of the desired cosmetic compositions. In this context, it is recommended to first carry out a sterilizing filtration on a filter with a pore diameter of 0.22 μm before packaging in bottles of appropriate volumes, then storage in a cold room, for example at 4° C.

[0060] On average, from 1 kg of ground longoza or Aframomum angustifolium seeds, about 3 to 3.5 kg of extract is obtained at 1% by weight of dry matter in butylene glycol, which corresponds to 30 to 35 grams of dry extract. The mass extraction yield obtained is on average 35% calculated as follows: mass yield=mass of dry extract contained in the final extract×100/mass of ground seeds introduced at the start.

EXAMPLE 2 ACCORDING TO THE INVENTION

[0061] Example of Preparation on an Industrial Pilot Scale of a Green Extract of Seeds of the Aframomum angustifolium or Longoza Plant, with a Glycol, for Example Butylene Glycol, According to the Invention

1) Pilot-Scale Tests

[0062] The pilot-scale tests are designed to process 10 kg per pilot-scale test of Aframomum angustifolium seeds from Madagascar according to 3 lots of seeds identified G1, G2 and G3 for Pilot-scale tests 1, 2 and 3, and adequate filtration systems described below.

2) Extraction Procedure According to the Invention in Plot-Scale Tests

[0063] The extract according to the invention is made with a polar solvent consisting of 100% butylene glycol from dry seeds of Aframomum angustifolium previously ground conventionally with a plant/solvent ratio (m/v) of 1/1.75 by first impregnating the plant by gentle maceration of 12 to 15 h at room temperature in butylene glycol to make the molecules of interest more accessible and available. A first filtration step is carried out to obtain a filtrate 1.

[0064] A second extraction is carried out at 55° C. on the cake for 3 h, then filtration is carried out on a filter with a porosity of 20-25 μm to obtain a filtrate 2. The two filtrates 1 and 2 obtained are combined and filtered again on a 0.5 to 0.1 μm filter.

[0065] The resulting solution is adjusted with butylene glycol to be 1% dry extract.

[0066] The result is a clear orangish glycolic solution.

[0067] The 3 extracts of the 3 lots G1, G2 and G3, obtained with the 3 pilot-scale tests, are named E1, E2 and E3.

[0068] On average, 10 kg of ground seeds produces about 30 to 35 kg of extract at 1% DM in butylene glycol (which corresponds to 300 to 350 g of dry extract).

[0069] The mass extraction yield obtained is on average 35%. It is calculated as follows:

mass yield=mass of dry extract contained in the final extract×100/mass of ground seeds introduced at the start.

3) Physicochemical Analyses of Extracts

[0070] 3-1) Comparable Dry Weight (DM): 0.96 to 1% (9.6 g/L to 10.0 g/L) pH In the range of 6.6; Comparable color: 7.8 to 7.9 (Gardner) Lab values 87.8-88.9; −3.3 to −2.5; 67.7 to 69.3.

[0071] 3-2) Total polyphenols: European Pharmacopoeia method 2.8.14 The test dose Is 0.05 g per sample. The reading is made at 750 nm in the visible.

TABLE-US-00001 Concentration (g/L) in gallic acid equivalent E1 0.012 E2 0.017 E3 0.016

[0072] Contents between 0.012 and 0.016 g/L are obtained. It can be observed that the contents are slightly higher for the extracts used with the E2 and E3 seed extracts.

4) Determinations of Total Sugars

[0073] Anthrone-sulfuric acid assay method. The reading is taken at 620 nm.

TABLE-US-00002 Content (%) in glucose equivalent E1 5.9 E2 7.0 E3 8.1

[0074] The contents are slightly higher for the extracts used with the E2 and E3 seed extracts.

Comparative Example 3 Between the Process According to the Invention of Example 2 Obtaining a Green Extract with Butylene Glycol from Seeds of the Aframomum angustifolium or Longoza Plant, According to the Invention and the Process According to Example 1 of U.S. Pat. No. 7,381,436 Obtaining an Alcohol/Water Extract 70/30 by Volume, from Seeds of the Aframomum angustifolium or Longoza Plant

[0075] 3-1) To carry out this comparative test, the following 2 samples were used: [0076] a) a comparative lot of seeds of the Madagascar Aframomum angustifolium or longoza plant called lot SX000HH which is subjected to the extraction process described in Example 1 of U.S. Pat. No. 7,381,436 which uses as extraction solvent a hydroalcoholic ethanol/water mixture 70/30 by volume and the extract is called extract lot SX000HH. [0077] b) the green extract E2 according to Example 2 of the above invention
3-2) Determination of the Anti-Aging Effect of the Green Extract E2 of Example 2 of Longoza (Aframomum angustifolium) on Normal Human Keratinocyte (NHK) Cells by Action on the Synthesis of ATP Synthase Subunit 6 by Mitochondria.

1. Introduction

[0078] A study on the transcriptional effects of butylene glycol green extract of longoza (Aframomum angustifolium) from Example 1 compared with the previous hydroalcoholic ethanol/water 70/30 volume extract according to Example 1 of the U.S. Pat. No. 7,381,436B2 was evaluated using the TaqMan low-density array (TLDA) technology. Using this technology, the modulation of the expression of 96 genes encoding specific proteins is studied in response to treatment of normal human keratinocytes.

[0079] After 24 hours of treatment, the cell monolayers were lysed to recover the RNAs. These RNAs were quantified and normalized to perform reverse transcription. The cDNAs obtained were then prepared for analysis on a NHF, NHK and NHM TLDA microfluidic card, 96 “chronobiology profile” genes.

2. Materials & Methods

2.1. Preparation of Tested Active Agents and Active Agent Controls

[0080] Active agent 1 according to the invention: 100% butylene glycol extract GREEN E2 from Madagascar longoza seeds according to Example 2 above.

[0081] Comparative active agent 2: extract with an EtOH/H.sub.2O 70/30 v/v mixture of Madagascar longoza seeds according to Example 1 of U.S. Pat. No. 7,381,436B2 and Example 3-1 a) above.

[0082] The final concentrations tested are 0.0125% by weight for Active agent 1 and Active agent 2.

2.2. Cell Treatments

[0083] The study was performed on normal human keratinocytes from abdominoplasty from a 31-year-old female donor (code 1477). Cells were first seeded in complete Epilife medium (HKGS supplement) at passage 4 (P4), at a density of 125 000 cells per well, in 12-well plates. The cells were then treated with 0.0125% Perle de Madagascar Green and 0.0125% Perle de Madagascar as well as their respective excipient control (BG 100% and EtOH/H.sub.2O 70/30 v/v).

[0084] After 4 hours of treatment, the cells are recovered in order to extract the total RNA.

2.3. TaqMan Low-Density Array (TLDA) Technique

2.3.1. Total RNA Extraction Using Microlab STAR (Hamilton)

[0085] The cell culture medium is removed, and 250 μL of RLT lysis buffer (provided in the Nucleospin RNA trace kit, (product code 740709, lot number 1412/001, Macherey-Nagel) is added. The cells are scraped with a cell scraper and then the cell lysate, recovered in a 1.2 mL deep-well (provided in the Nucleospin RNA kit). Total RNA is extracted according to previously developed protocols (NS_96RNA_KHN or NS_96RNA_FHN.dws). The total RNA solutions obtained are assayed, and their quality checked, using a microplate reader, the SPECTROstar Nano (BMG Labtech) coupled to the Microlab STAR. This device is connected to the computer controlling the robotic platform and has specific software for analyzing the results (MARS software). The technique requires a 384-well microplate (384 micro-clear plate black LoBase, product code: 788876 lot: 1305055 Greiner), a positive control (RNA 250, product code: AM7155, lot: 1301028 (NHF), lot: 1208025 (NHK), lot: 1208026 (NHM), Thermofisher) to validate the pipetting performed by the robot as well as the values generated by the SPECTROstar Nano reader.

2.3.2. Synthesis of Complementary DNA

[0086] The reverse transcription (RT) kit used is the High-Capacity Reverse Transcription Kit (product code: 4368813, lot: 00288647; Thermo Fisher) and was used according to the protocol provided: 500 ng total RNA is diluted in water to a final volume of 25 μL. It is then incubated for 10 minutes at 25° C. then 2 hours at 37° C. in the presence of 25 μL of High-Capacity Reverse Transcription Kit 2× reaction mixture previously prepared as shown below. The different incubations are carried out in the TRobot (Biométra).

TABLE-US-00003 TABLE 2 Preparation of the High-Capacity Reverse Transcription Kit 2X reaction mixture for 1 reaction Random RNase RT primer out Reagents buffer dNTP probe enzyme RT H.sub.2O Volume 5 μL 2 μL 5 μL 0.5 μL 2.5 μL 10 μL

2.3.3. PCR-TaqMan Low-Density Array

[0087] 15 μL of each RT is mixed with 60 μL water then 75 μL TaqMan Gene Expression Master Mix (product code: 4369016, lot: 1507235; Thermo Fisher) is added. After homogenization, 100 μL is deposited on the 96-gene chronobiology profile microfluidic cards (lot B1885 (NHF), lot B1884 (NHK) and lot B1942 (NHM)), which are centrifuged and then sealed.

[0088] The CD, corresponding to the profile of the genes deposited on the plates, is loaded into the SDS 2.4 software, which thus makes it possible to specify the location of each gene on the card. The control genes (called “endogenous” genes in the software) to be used for normalization of results are indicated before the PCR is started. PCR is performed according to the protocol provided by Applied Biosystems in the ABI Prism 7900HT Sequence Detection System. The qPCR steps are 2 min at 50° C., 10 min at 94.5° C. then 30 min at 97° C. and 1 min at 59.7° C. for 40 cycles.

2.3.4. Statistical Analysis of Results

[0089] Real-time quantitative PCR can be exploited if its efficiency is between 90% and 110%.

[0090] In the RT-PCR/TLDA method, quantitation is performed using the comparative method of ΔCt, i.e., relative quantitation (RQ). This method determines the Ct of each gene on the card using the RQ Manager software which takes into account the background noise for each gene. This Ct is normalized to the Ct of a housekeeping gene (in our study beta-2-microglobulin) which is studied in parallel on the card. This gene being invariant in the cells, its variations reflect the variability of the experiment (total RNA assay, pipetting, reverse transcription step, deposits in the microfluidic card, quality of the card, PCR in the apparatus). With this RQ Manager software, we obtain the RQ (Relative Quantitation) values which correspond to the amplitude level (x times greater or smaller than the control) of the expression compared with our control. The latter can be the untreated control or the excipient. The RQ is obtained by the following calculation (the value of the untreated being equal to 1):

RQ=2−ΔΔCt=2−(ΔCt treated−ΔCt untreated)

ΔCt treated=Ct treated target gene−Ct treated housekeeping gene
ΔCt untreated=Ct untreated target gene−Ct untreated housekeeping gene

[0091] For this method to be reliable, it is imperative that the amplification efficiency of the gene of interest and the housekeeping gene be the same. This was verified during the development process prior to routine use of the plates.

[0092] Student's t-test will be used to evaluate statistically significant variations in transcriptional activity. Each condition is performed in triplicate (3 untreated and 3 treated under the same conditions). Fisher's F-test is first applied by comparing the two data matrices. When the value is greater than α=0.05 then the variance of Student's t-test is 2, when Fischer's F-test is less than α=0.05 then the variance will be equal to 3. The transcriptional variations selected will be those corresponding to a Student's t-test lower than α=0.05.

TABLE-US-00004 TABLE 3 Coding table RQ < 0.5 (F) 0.5 < RQ < 0.75 (E) 0.75 < RQ < 1 (D) 1 < RQ < 1.5 (C) 1.5 < RQ < 2 (B) RQ > 2 (A)

[0093] A criterion taking into account the expected effect (+ for an Increase, − for an inhibition) allows a first interpretation of results to be made.

[0094] All these data allow us to note the extracts in relation to each other since the Excel macro indicates the number of genes modulated in the expected direction/number of modulated genes in total. In the context of a screening, this score can be used as a selection criterion.

[0095] Results relating to the effect of longoza green butylene glycol extract E2 according to the invention of Example 2, compared with longoza prior to the ethanol/water 70/30 vol/vol hydroalcoholic mixture of U.S. Pat. No. 7,381,436B2 example 1, on normal human keratinocytes (NHK) after 4 h.

TABLE-US-00005 TABLE 4 Effect of longoza green E2 and longoza on normal human keratinocytes L Green E2 Longoza 0.0125% 0.0125% 4 h 4 h ATP synthase 6 encoded MT + 2.697 (A) — in mitochondria

[0096] The longoza green E2 according to Example 2 of the invention increases in an unexpected manner by a coefficient of +2.697 the synthesis of ATP synthase subunit 6 by NHK mitochondria.

[0097] In conclusion of the above, thanks to the above test on NHK, it was possible to demonstrate the significantly repairing effect of the mitochondrial dysfunction of the green butylene glycol extract of longoza according to the invention, which combats the senescence of mitochondrial cells and therefore combats skin aging.

[0098] Thus, the invention can be used as a cosmetic agent for the preparation of a cosmetic composition for topical use to preserve the mitochondria and to combat skin aging.

[0099] Various examples of formulations of cosmetic compositions are given below by way of illustration and therefore in no way limit the scope of the invention.

EXAMPLE 4 ACCORDING TO THE INVENTION

Cosmetic Gel for Improving the Firmness of the Face

[0100]

TABLE-US-00006 butylene glycol 100% green extract of longoza 2% obtained in Example 1, g) glycol 3% commercially available AMPS polymer, 3% (trade name Sepigel 305 ®) hydrogenated castor oil (Cremophor CO-60 ®) 2% polyethylene glycol 1.5%.sup.  preservative 0.5%.sup.  perfume concentrate 0.3%.sup.  UV filter (benzophenone-4) 1% water, q.s, 100%

[0101] This gel can be applied to the face once a day for several weeks in order to improve the firmness of the facial skin, especially on areas with wrinkles, thus observing at the end of this treatment, a restoration of suppleness with a clear effect of skin rejuvenation and attenuation or disappearance of these wrinkles.

EXAMPLE 5 ACCORDING TO THE INVENTION

Anti-Wrinkle Day Cream in the Form of an Emulsion

[0102]

TABLE-US-00007 butylene glycol 100% green extract E1 of longoza .sup. 2% obtained in Example 2 steareth-21 (Brij 721) 2.5% glyceryl stearate (Tegrin) 1.1% stearyl alcohol .sup. 5% glycerol tricaprylate/caprylate 12.5%  butylene glycol .sup. 3% glycerin .sup. 2% preservative 0.5% perfume concentrate 0.5% UV filter (octyl methoxycinnamate) 7.5% water, q.s. 100%

EXAMPLE 6 ACCORDING TO THE INVENTION

Anti-Wrinkle Tonic Lotion

[0103]

TABLE-US-00008 butylene glycol 100% green extract E2 of longoza 2% obtained in Example 2 butylene glycol 3% EDTA 0.1%.sup.  solubilizer 1% perfume concentrate 0.3%.sup.  ethanol 5% UV filter (benzophenone-4) 0.13%   water, q.s. 100%

EXAMPLE 7 ACCORDING TO THE INVENTION

Ultraviolet Protective Make-Up Powder for the Face

[0104]

TABLE-US-00009 butylene glycol 100% green extract E3 of longoza 0.25%.sup.  obtained in Example 2 talcum 17% mica 20% sericite 20% pigments  8% organic powder (nylon) 20% silica 8.75%.sup.  mineral oil or silicone  3% UV filter (octyl methoxycinnamate)  3%

EXAMPLE 8 ACCORDING TO THE INVENTION

Anti-Wrinkle Treatment Foundation

[0105]

TABLE-US-00010 butylene glycol 100% green extract E2 of longoza .sup. 2% obtained in Example 2) 4-polyglyceryl isostearate and cetyl dimethicone 5.1% copolyol and hexyl laurate cyclopentasiloxane and cyclohexasiloxane 5.0% cetyl dimethicone 1.0% caprylic/capric triglycerides 2.2% octyl stearate 1.4% mineral oil 3.5% hydrogenated castor oil 1.2% beeswax 0.8% polymethyl methacrylate 1.1% iron oxides 0.45%  titanium dioxides 5.2% NaCl 0.6% perfume concentrate 0.1% UV filter (octyl cinnamate) .sup. 3% water, q.s, 100%