ORGANIC COMPOUNDS

Abstract

A cosmetic ingredient comprising crenatoside and acteoside when applied to the skin is useful to provide cosmetic and perfumery benefits.

Claims

1. A cosmetic ingredient comprising crenatoside and acteoside.

2. The cosmetic ingredient according to claim 1 in the form of a plant extract comprising crenatoside and acteoside.

3. The cosmetic ingredient according to claim 2, comprising the extract and a hydric organic solvent.

4. The cosmetic ingredient according to claim 3, wherein the hydric organic solvent includes-glycerol, a glycol or 1,3-propanediol.

5. A formulation adapted for topical application to the skin comprising the cosmetic ingredient according to claim 1.

6. The formulation according to claim 5, comprising a perfume composition containing at least one perfume ingredient.

7. The formulation according to claim 5, selected from the group consisting of: a cosmetic cream, cosmetic lotion, cosmetic body spray, cosmetic serum, a deodorant or antiperspirant, or a fine fragrance.

8. A method of modulating the evaporation of a perfume composition applied to the skin, the method comprising the step of: applying simultaneously, sequentially or separately the cosmetic ingredient according to claim 1 to the skin.

9. A method of improving the efficacy and/or long-lastingness of a deodorant composition, comprising the step of adding the cosmetic ingredient according to claim 1 to the deodorant composition.

10. A method of facilitating the transition from an antiperspirant to a deodorant composition by providing a deodorant composition comprising the cosmetic ingredient according to claim 1.

11. A deodorant composition or an antiperspirant composition which comprises the cosmetic ingredient of claim 1, and further wherein the deodorant composition or antiperspirant composition comprises no aluminium salts or zirconium salts.

12. The cosmetic ingredient according to claim 3, wherein the hydric organic solvent is 1,3-propanediol.

13. The cosmetic ingredient according to claim 12, wherein the plant extract contains at least 15 wt %, of a mixture of crenatoside and acteoside, based on the total weight of the dry matter of the plant extract.

14. The cosmetic ingredient according to claim 13, wherein the mixture comprises about 20 wt % to about 60 wt % of crenatoside and about 80 wt % to about 40 wt % of acteoside, based on the total weight of said mixture.

15. The cosmetic ingredient according to claim 12, wherein the cosmetic ingredient of the present invention contains at least 15 wt % of each of crenatoside and acteoside, based on the total weight of the dry matter of the plant extract.

16. A topical treatment formulation for application to the skin comprising the cosmetic ingredient of claim 12.

17. The topical treatment formulation according to claim 16, comprising a perfume composition containing at least one perfume ingredient.

18. The topical treatment formulation according to claim 16, which is selected from the group consisting of: a cosmetic cream, cosmetic lotion, cosmetic body spray, cosmetic serum, a deodorant or antiperspirant, and a fine fragrance.

19. A method of modulating the evaporation of a perfume composition applied to skin, the method comprising the step of applying simultaneously, sequentially or separately the cosmetic ingredient according to claim 12 to the skin.

20. A method of improving the efficacy and/or long-lastingness of a deodorant composition, comprising the step of adding the cosmetic ingredient according to claim 12 to the deodorant composition.

21. The method of claim 20, wherein the malodour reduction is improved and the fragrance intensity is increased.

22. A method of facilitating the transition from an antiperspirant to a deodorant composition by providing a deodorant composition comprising the cosmetic ingredient according to claim 12.

23. A deodorant or antiperspirant composition which comprises the cosmetic ingredient of claim 12, and further wherein the deodorant or antiperspirant composition excludes aluminum salts and zirconium salts.

24. A method of preparing the cosmetic ingredient according to claim 12, the method comprising the steps of: I) extracting a plant source containing the compounds crenatoside and acteoside in an extraction solvent, the extraction solvent being ethanol; II) filtering the extract to remove solids; III) carrying out a solvent-exchange step comprising mixing the extract with a hydric solvent, the hydric solvent being 1,3-propanediol, and removing the extraction solvent by evaporation; IV) optionally decolourizing the extract obtained in step III), using a suitable decolourizing agent; V) filtering the extract to remove solids; and VI) optionally adjusting the extract to the desired dry matter content by evaporation.

25. The method of claim 24, wherein the plant extract contains at least 15 wt %, of a mixture of crenatoside and acteoside, based on the total weight of the dry matter of the plant extract.

26. The method of claim 24, wherein the mixture comprises about 20 wt % to about 60 wt % of crenatoside and about 80 wt % to about 40 wt % of acteoside, based on the total weight of said mixture.

27. The method of claim 24, wherein the cosmetic ingredient contains at least 15 wt % of each of crenatoside and acteoside, based on the total weight of the dry matter of the plant extract.

Description

EXAMPLE 1

Preparation of Cosmetic Ingredient

[0087] Dried Orobanche rapum, collected manually in Lozére, France, was dried for 1 month in shady conditions before being ground into powder. 1 kg of Orobanche rapum powder was extracted in 6 kg of 96% ethanol in the absence of water. Thereafter, the solid material was separated from the liquid extract by filtration over a cellulosic filter (0.7 micron).

[0088] The filtrate was combined with an equal mass of 1,3-propanediol before the ethanol was removed by vacuum evaporation. The resultant extract in 1,3-propanediol was filtered over a charcoal filter. The decolourized extract was adjusted to 1 wt % dry matter and filtered on a 0.3 micron filter.

[0089] Analysis by 2D NMR identified two major peaks that representing 18.9 wt % (based on total weight of dry matter), which peaks correspond to crenatoside and acteoside.

EXAMPLE 2

Metagenomic Analysis

[0090] Clinical assessments of the cosmetic ingredient formed according to Example 1 were carried out on human volunteers.

[0091] A double blind and placebo-controlled clinical evaluation was carried out with 19 women (Age: between 18 and 50 years old, mean age: 39.5 years) having dry skin (corneometer value below 40 AU). All of the subjects participating in the study gave their informed consent signed at the beginning of the study. The measurements were done after 14 and 28 days of use with a test formula (set forth below) containing 0.5% of the extract obtained in accordance with Example 1. A placebo consisted of the same formulation (below) without the extract.

[0092] In this study, the evolution of skin metagenome was followed. The body area studied was the forearm.

[0093] The test formulation, in the form of a cream, contained the following ingredients:

[0094] WATER, CAPRIC/CAPRYLIC TRIGLYCERIDE, CETEARYL WHEAT STRAW GLYCOSIDES, CETEARYL ALCOHOL, EXTRACT OF EXAMPLE 1, PHENOXYETHANOL, DIMETHICONE, METHYL PARABEN, PROPYL PARABEN, ETHYL PARABEN, FRAGRANCE, HEXYL CINNAMAL, BUTYLPHENYL METHYLPROPIONAL, CITRONELLOL, ALPHA ISOMETHYL IONONE, HYDROXYISOHEXYL 3-CYCLOHEXENE CARBOXALDEHYDE, SODIUM HYDROXIDE.

[0095] A placebo formulation was identical to the test formulation (above) except that the extract of Example 1 was omitted.

[0096] Skin samples of cutaneous microflora were collected from the forearms of healthy volunteers (50 cm.sup.2) by a non-invasive swabbing method using sterile swabs moistened with a sterile solution of 0.15 M NaCl. Swabs were transferred at −20° C. and kept frozen until DNA extraction.

[0097] DNA extraction was performed using the PowerLyzer® PowerSoil® DNA Isolation Kit (MO BIO Labora-tories, Inc., Carlsbad, USA), with the following modifications. The tip of each swab was detached with a sterile surgical blade and transferred to a 1.5 mL tube to which 750 μL of Bead Solution had been added. The sample biomass was re-suspended by stirring and pipetting and the biological suspension was transferred to a bead beating tube. The remaining steps were performed according to the manufacturer instructions.

[0098] Sequencing and DNA analysis was carried out as described below.

[0099] 16S rRNA gene sequencing:

[0100] Sequencing was performed with the MiSeq device (Illumina, Inc., San Diego, Calif., USA) through a 500 cycles paired-end run, targeting the V3V4 16S variable regions using the following primers: 16S-Mi341F forward primer 5′-CCTACGGGNGGCWGCAG-3′ and 16S-Mi805R reverse primer 5′-GACTACHVGGGTATCTAATCC-3′, producing about 460 bp amplicons.

[0101] PCR1s were performed as follows: 8 μL of template DNA (0.2 ng) were mixed with 5 μL of each reverse and forward primer (1 μM), 5 μL of KAPA HiFi Fidelity Buffer (5×), 0.8 μL of KAPA dNTP Mix (10 mM each), 0.7 μL of RT-PCR grade water (Ambion), and 0.6 μL of KAPA HiFi hotstart Taq (1 U/μL), for a total volume of 25 μL. Each amplification was duplicated, and duplicates were pooled after amplification. PCR1 cycles consisted of 95° C. for 3 min and then 32 cycles of 95° C. for 30 s, 59° C. for 30 s, and 72° C. for 30 s, followed by a final extension at 72° C. for 3 min, with a BioRad CFX1000 thermocycler. Negative and positive controls were included in all steps to check for contamination. All duplicate pools were controlled by gel electrophoresis, and amplicons were quantified using fluorometry.

[0102] Libraries ready for analysis were then produced following the Illumina guidelines for 16S metagenomics libraries preparation. Briefly, the PCR1 amplicons were purified and controlled using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, USA). To enable the simultaneous analysis of multiple samples (multiplexing), Nextera® XT indexes (Illumina) were added during PCR2 using between 15 to 30 ng of PCR1 amplicons. PCR2 cycles consisted of 94° C. for 1 min and then 12 cycles of 94° C. for 60 s, 65° C. for 60 s, and 72° C. for 60 s, followed by a final extension at 72° C. for 10 min. Indexed libraries were purified, quantified and controlled using an Agilent 2100 Bioanalyzer. Validated indexed libraries were pooled in order to obtain an equimolar mixture.

[0103] The run (500 cycles) was achieved on MiSeq sequencer (Illumina) using the MiSeq Reagent Kit v3 600 cycles (Illumina). The sequencing run produced an output of 25 million of paired-end reads of 250 bases, i.e. up to 6 Gigabases. The libraries and the MiSeq run were performed by Libragen, at the GeT-PlaGe platform (INRA, Auzeville, France).

[0104] After MiSeq run, raw data sequences were de-multiplexed and quality-checked to remove all reads with ambiguous bases. Indexes and primer sequences were then trimmed, and the forward and reverse sequences were paired. The paired-sequences were then treated using in-house pipeline to remove chimeras and reads with PCR errors and to split sequences into Operational Taxonomic Unit (OTU) at a 1% dissimilarity level. Good quality binned paired-sequences were mapped to the SILVA SSU Ref database (Release 123; https://www.arb-silva.de/) for taxonomic assignation. Data were then normalized, and compared using White's non-parametric test (White et al., 2009).

[0105] The results are shown in the following two tables.

TABLE-US-00001 TABLE 1 Forearm microbiota phyla composition after 14 days of treatment with Vehicle and Vehicle + Active Vehicle Vehicle + Active D0-Active: D14-Active: D0-Active: D14-Active: mean rel. mean rel. mean rel. mean rel. Phylum frequ. (%) frequ. (%) p-value frequ. (%) frequ. (%) p-value Actinobacteria 38.71 34.42 0.387 38.05 33.88 0.443 Proteobacteria 26.78 22.27 0.372 27.25 25.73 0.775 Firmicutes 26.19 37.00 0.025 26.10 29.47 0.486 Bacteroidetes 6.18 3.61 0.144 6.62 3.94 0.176 Cyanobacteria 1.15 1.91 0.496 1.36 6.04 0.272 Soccharibacteria 0.14 0.09 0.092 0.09 0.08 0.852 Chloroflexi 0.11 0.09 0.794 0.07 0.14 0.061

[0106] Table 1 lists the main phyla (in terms of relative abundance) found in the study. Table 1 also demonstrates that the microbiota evolves differently depending on whether skin was treated with the test formulation (“Vehicle+Active”) or the placebo (“Vehicle”) formulation. At the phylum level, skin treated with the placebo formulation exhibited a significant increase of Firmicutes between D0 and D14, whereas the levels of other phyla remained substantially stable. On the other hand, for skin treated with the test formulation, levels of all phyla, including Firmicutes, remained substantially stable.

[0107] It can be concluded that the cosmetic ingredient stabilises the microbiota and maintains the essential balance of the skin microflora.

TABLE-US-00002 TABLE 2 Comparison of the relative abundances of Finegoldia magna at 0 and D14 Vehicle + Active 0.5% Vehicle D0-Active: D14-Active: D0-Active: D14-Active: mean rel. mean rel. mean rel. mean rel. Genus frequ. (%) frequ. (%) p-value frequ. (%) frequ. (%) p-value Finegoldia 3.1 1.3 0.043 1.9 2.1 0.741

[0108] Table 2 shows the levels of Finegoldia magna on skin treated with test formulation and placebo (Vehicle) over a 14-day period.

[0109] Finegoldia magna is a species that is a normal inhabitant of human skin and is most frequently Gram positive cocci isolated from infected lesions. F. magna is implicated in mono- and poly-microbial infections of skin, bone, heart and meninges. A case of toxic shock syndrome caused by F. magna has also been reported in the literature.

[0110] Results of the study show that skin treated with the test formulation (“Active 0.5%”) exhibits a significant reduction in the relative abundance of Finegoldia over the 14 day test period (−58%). On the other hand, when skin is treated with the placebo formulation (“Vehicle”), the level of Finegoldia remains stable. These finding suggest that the cosmetic ingredient inhibits the growth of opportunistic pathogens.

EXAMPLE 3

Deodorant Composition

[0111] In order to assess the effect of the cosmetic ingredient of the present invention on deodorant efficacy, a double blind study on 10 volunteers was conducted. All volunteers were antiperspirant users.

[0112] The volunteers had to stop using their usual antiperspirant. Instead, they applied a first test deodorant composition on their left underarm and a second test deodorant composition on their right underarm each morning.

[0113] At the end of a five-day test period, the volunteers made a self-assessment of the malodour and the fragrance for each side.

[0114] Both test deodorant compositions were provided in the form of a spray and contained the following standard deodorant base:

TABLE-US-00003 1 wt % Octyl Dodecanol 36 wt %  Ethyl alcohol 96% 2 wt % Propylene Glycol 1 wt % Perfume 60 wt %  Propellant (Isobutane, Propane)

[0115] The above ingredients were mixed without the propellant and supplemented with 1.75 wt % of a fragrance composition. For one of the two test deodorant compositions, 0.5 wt % of the cosmetic ingredient of the present invention was added. The mixtures were filled into a spray container and charged with the propellant.

[0116] The self-assessment showed that 9 out of the 10 volunteers found that the test deodorant composition containing the cosmetic ingredient of the present invention provided a better malodour reduction and an increased fragrance intensity.

[0117] Thus, the cosmetic ingredient of the present invention is able to improve the long-lastingness of a deodorant composition.