FR2871691A1 - Cosmetic composition, useful to e.g. prevent/treat skin inflammations and cellulite, comprises an extract of leaves of Albizia adanthifolia - Google Patents

Abstract

Cosmetic composition (I) comprises an extract (A) of leaves of Albizia adianthifolia. ACTIVITY : Dermatological; Antiinflammatory. MECHANISM OF ACTION : Matrix metalloprotease inhibitor; Melanin synthesis inhibitor; Prostaglandin E2 inhibitor; Cyclooxygenase inhibitor; Phospholipase A2 inhibitor; Elastase inhibitor. The ability of (A) to inhibit matrix metalloprotease-3 (MMP-3) was tested using modified method of Nagase and fields (J Biol Chem Vol. 269, 33, 20952-20957, 1994). The results showed that (A) allowed inhibition until 29% of the degradation activity of MMP-3.

Description

2871691 1

  The present invention relates to a composition based on an extract of Albizia adianthifolia leaves, useful in particular for protecting the skin from external aggressions and to fight against skin aging, reduce inflammatory processes, ensure the whitening of the skin and generate an action. slimming.

  Albizia adianthifolia (synonym Albizzia adianthifolia) is a plant species native to East Africa of the family Leguminosae, represented by large trees measuring between 10 and 25 meters and whose leaves are deciduous and bipinnate.

  Several species of Albizia have been described to contain alkaloids, triterpenes, triterpenic saponosides, especially in roots and bark, and flavonols.

  Particular saponins have been identified in the roots of Albizia adianthifolia (Haddad et al., J. Nat Prod, 2003, 66, 372-377).

  In addition, the traditional use of certain extracts of Albizia adianthifolia has been cited in the literature, including the use of extracts obtained from roots, useful for the treatment of skin diseases, or as a remedy against intestinal parasites or snake bites (Mazzanti et al., Fitoterapia, 1983, 65, 275).

  Korean patent application KR2003064059 for its part describes a cosmetic composition comprising the synergistic combination of an extract of Albizia julibrissin and an extract of Paeonia suffruticosa, which has anti-aging and antioxidant properties. Application KR2002080657 describes a cosmetic composition comprising an extract of Albizia julibrissin, useful for its antioxidant properties.

  The inventors have now demonstrated that an extract of Albizia adianthifolia leaves has particularly interesting cosmetic properties.

  The present invention therefore relates to a cosmetic or pharmaceutical composition comprising, in addition to a cosmetically or pharmaceutically acceptable vehicle, an extract of Albizia adianthifolia leaves.

  By cosmetically or pharmaceutically acceptable vehicle is meant a suitable vehicle for use in contact with human and animal cells, in particular the cells of the epidermis, without toxicity, without irritation, without undue allergic response and the like, and proportioned at a reasonable risk / benefit ratio.

  The Albizia adianthifolia leaf extract can be obtained by extraction with a solvent selected from methanol, ethanol, water, ethyl acetate and a mixture of these solvents or from propylene glycol, butylene glycol mixed or not with water. The solvent mixtures can be made in any possible proportion.

  According to one embodiment of the invention, the extract of Albizia adianthifolia leaves is obtained after grinding the sheets in a solvent and hot extraction, between 40 and 60 C, in the same solvent for one to two hours. The extract is then filtered and dried, for example by spraying or lyophilization.

  It should be noted that the extract may be concentrated by evaporation under vacuum or that a particular fraction of the extract may be purified or concentrated, in particular by liquid / liquid extraction or by preparative chromatography.

  According to a preferred embodiment of the invention, the extract of Albizia adianthifolia leaves can be obtained by hydroalcoholic extraction, preferably in an ethanol / water mixture in proportions of 70/30 by volume.

  Preferably, the compositions according to the invention may comprise from 0.0001 to 0.1% of said plant extract expressed by weight relative to the total weight of the composition.

  The administration of a composition according to the invention is preferably carried out topically.

  The composition is intended more particularly for the treatment of skin and mucous membranes and may be in the form of cream, oil, milk, solution, gel, spray, lotion or suspension.

  Even more preferably, the compositions of the invention are cosmetic compositions in the form of oil-in-water or water-in-oil emulsion, or multiple emulsion, microemulsion, aqueous gel, or aqueous-alcoholic, cream, oil, aqueous or hydroalcoholic lotion.

  These compositions are particularly advantageous for combating skin aging, especially photoaging, to whiten the skin, to reduce skin inflammation, to produce a thinning and / or fight against cellulite.

  The skin aging is caused on the one hand by a decrease in collagen synthesis and on the other hand by a degradation of the dermal matrix due to the increase in the induction of MMPs or metalloproteases of the matrix. These enzymes are indeed increasingly expressed during aging and during exposure to certain stresses, including UV radiation or pollutants.

  There are several distinct matrix metalloproteases. MMP 1 or collagenase-1 acts at the level of the deep dermis and degrades collagens I and III. MMP 2 or gelatinase A is present at the dermal-epidermal junction and degrades collagens I, III and VII, as well as elastin. Mention may also be made of MMP 3 and elastase, which are enzymes for degrading components of the dermal extracellular matrix.

  The extract of leaves of Albizia adianthifolia is thus useful to fight against skin aging. In particular, this plant extract is capable of stimulating the proliferation of dermal cells and protecting them with respect to the various matrix metalloproteases by reducing their induction under UV stress conditions and by inhibiting their activity.

  An object of the invention is therefore the use of a cosmetic composition comprising an extract of Albizia adianthifolia leaves to fight against skin aging, especially photoaging, and protect the skin from external aggression.

  Albizia adianthifolia leaf extract is also useful as a whitening and depigmenting agent. This extract was effective both on skins, called high phototype skins and on light skin with pigmentation tasks, including age spots. It has thus been shown that this extract inhibits the synthesis of melanin by melanocytes by decreasing the activity of an enzyme, tyrosinase, which is essential for the synthesis of this pigment.

  An object of the invention is therefore the use of a cosmetic composition comprising an extract of Albizia adianthifolia leaves for bleaching and / or depigmentation of the skin.

  The extract of Albizia adianthifolia leaves is also useful as a slimming agent. The inventors have demonstrated the lipolytic action of the extract of Albizia adianthifolia leaves on an adipocyte conversion model in which the triglycerides were labeled and assayed. By adipocyte conversion, it is meant the transformation of the immature adipocyte into a differentiated mature adipocyte. Adipocytes are cells located in the hypodermis and responsible for storing energy in the form of triglycerides. In their cytoplasm, lipogenesis is the esterification of fatty acids and glycerol into triglycerides. The reverse hydrolysis reaction corresponds to lipolysis. The extract significantly reduced the level of triglycerides present in the differentiated adipocytes compared to untreated controls and compared to caffeine which is known for its strong lipolytic action.

  An object of the invention is therefore the use of a cosmetic composition comprising an extract of Albizia adianthifolia leaves as a slimming cosmetic composition.

  The extract of Albizia adianthifolia leaves is also useful as an anti-inflammatory agent. It has thus been shown that this plant extract inhibits the release of various pro-inflammatory mediators, in particular prostaglandins of the E2 type, by the skin cells, in particular by human keratinocytes. The inventors have furthermore demonstrated that Albizia adianthifolia leaf extract inhibited the activity of other inflammatory mediators such as phospholipase A2 and cycloxygenase. Arachidonic acid is formed from the phospholipids of cell membranes under the action of phospholipases such as phospholipase A2. An enzyme, cyclooxygenase, transforms arachidonic acid into prostaglandins, particularly of type E2 (PGE2). These prostaglandins are lipid mediators, which play an important role in the constitution of edema, causing vasodilation and increasing vascular permeability.

  An object of the invention is therefore the use of an extract of Albizia adianthifolia leaves for the preparation of a cosmetic or pharmaceutical composition useful for the prevention or treatment of inflammations, in particular cutaneous inflammations.

  The authors of the present invention propose to take advantage of this anti-inflammatory property associated with the lipolytic action mentioned above, to combat cellulite. An object of the invention is therefore the use of an extract of Albizia adianthifolia leaves for the preparation of a pharmaceutical or cosmetic composition for the prevention or treatment of cellulite.

  The following examples illustrate the invention without limiting its scope.

EXAMPLE 1

  Preparation of Albizia Adianthifolia Leaf Extract In the examples below, an extract prepared as follows was extracted from Albizia adianthifolia leaves. The leaves are crushed and mixed with an ethanol / water solvent mixture, in proportions 70/30 (by volume). The extraction is carried out hot between 40 and 60 C for 1 to 2 hours.

  The extract obtained is then filtered and dried.

  This extraction makes it possible to obtain an extract whose effectiveness has been tested with respect to various cosmetic properties.

EXAMPLE 2

  Protection! Dermal prevention and anti-aging A) Enzymatic tests 1. Inhibition of MMP3 The Albizia adianthifolia leaf extract was tested according to the modified method of Nagase and Fields (J Biol Chem Vol 269, 33, 20952- 20957, 1994).

  In this technique, the NFF2 substrate (Mca-Arg-Pro-Lys-Pro-Tyr-Ala-NvaTrp-Met-Lys (Dnp) -NH2) is contacted with the type 3 matrix metalloprotease and converted into a peptide motif. recognizable in fluorometry. This formation, which is dependent on the activity of the enzyme, is monitored according to the extract to be tested.

  (Mca = 7-amino-methyl-coumarin, Nva = norvaline, Dnp = 2,4 dinitrophenol) The results of these experiments show that Albizia adianthifolia leaf extract inhibits up to 29% of degradation activity of the MMP 3 and therefore has a protective activity vis-à-vis the damage of the dermis due to the MMP 3.

  2. Inhibition of elastase The Albizia adianthifolia leaf extract was tested according to the method adapted and provided by SANOFI PHARMA (J Pharmacol Exp Ther., 260 (2): 809-16, 1992).

  In this technique, the substrate, N-succinyl-ala-ala-pro-leu-p. Nitroanilide is brought into contact with elastase and forms an acyl-enzyme by releasing p. nitroaniline which absorbs at 386 nm. The enzymatic activity is defined with respect to the slope of the release kinetics of p. nitroaniline.

  Leaf extract of Albizia adianthifolia is considered very active in the inhibition of elastase. The IC50 of the extract is 0.0314 mg / mL. The extract therefore has a dermal protection activity against degradation that may be caused by elastase.

  B) Cellular Assays: Stimulation of Fibroblast Proliferation and Inhibition of MMP1 and 2 Induction a) Materials and Methods The assay is performed from culture of normal human dermal fibroblasts. The test extract is brought into contact with fibroblasts under different conditions, namely before, during and after UV stress or during the entire manipulation. Thirty hours of incubation after UV stress, cell supernatants are recovered and centrifuged. MMPs 1 and 2 are assayed via an ELISA technique (Tables 1 and 2) and cell viability is estimated by the thiozolyl blue technique, MTT (Table 3).

  b) Results Table 1: Determination of MMP 1 (ng / cell unit / mL) Treatment Stress from 0% inhibition to% inhibition stress at 20 J / cm 2 OJ / cm 2 20 J / cm 2 J / cm 2 Before stress UV 28 -14 20 31 During UV stress 15 40 19 36 After UV stress 0 100 4 86 Set 0 100 0 100 Without treatment 25 (= 100%) 29 (= 100%) 2871691 7 Table 2: Determination of MMP 2 (ng cell unit / mL) Treatment Stress% inhibition at% inhibition stress at 0 J / cm2 OJ / cm2 20 J / cm2 20J / cm2 Before UV stress 25 27 72 0 During UV stress 43 -23 77 -7 After UV stress 30 15 34 52 Together 27 23 48 33 Without treatment 35 (= 100%) _72 (= 100%) The addition of Albizia adianthifolia leaf extract has a significant influence on inhibition MMP 1 and MMP 2 released during a UV stress.

  Table 3: Effect on cell viability Treatment Stress of 0 J / cm2 Stress of 20 J / cm2 Before stress UV -32% -5% During stress UV -11% - 9% After stress UV + 7% + 53% Together + 29% + 44% Without treatment 100% 100% The addition of the extract in the culture medium of human fibroblasts, after the UV stress, makes it possible to increase by 53% the viability of the cells.

  The extract of Albizia adianthifolia leaves on the one hand to stimulate the proliferation of dermal fibroblasts and to limit their production of MMP 1 and 2 under stress conditions.

EXAMPLE 3

  Inhibition of Cycloxygenase The Albizia adianthifolia leaf extract was tested according to the modified method of Mitchell and Akarasereenont (Natl Acad Sci USA 90 (24): 11693-11697, 1993).

  In this technique, arachidonic acid is contacted with cycloxygenase and converted to prostaglandin E2. This formation depending on the activity of the enzyme is monitored as a function of the concentration of the extract.

  The results of these experiments show that the extract of Albizia adianthifolia can inhibit up to 41% of the activity of cycloxygenase 2 (inducible form) and therefore has an anti-inflammatory activity.

  2. Inhibition of phospholipase A2 Albizia adianthifolia leaf extract was tested according to the modified method of Katsumata and Gupta (Anal Biochem 154: 676-681, 1986).

  In this technique, tritiated phosphatidylethanolamine is contacted with phospholipase A2 and converted to arachidonic acid. The formation of arachidonic acid dependent on the disappearance of the labeled substrate and dependent on the activity of the enzyme is monitored as a function of the concentration of the extract.

  The results of these experiments show that Albizia adianthifolia leaf extract inhibits up to 36% of the activity of phospholipase A2 and therefore has anti-inflammatory activity.

  B) Tests on cells: inhibition of the release of PGE2 a) materials and methods The anti-inflammatory activity was evaluated on human keratinocytes subjected to a phorbol ester, here phorbol myristate acetate (PMA) or known ultraviolet rays to induce an inflammatory reaction, and in particular to stimulate the production of prostaglandins type E2 or PGE2.

  The keratinocyte cells (HaCat) are placed in the presence of an extract of Albizia adianthifolia leaves treated with PMA for 24h at 37 ° C. or subjected to UV irradiation under the following conditions UVA at 25J / cm 2 and UVB at 250 mJ / cm 2 , during 1:30.

  After 24 h, the inflammatory state is demonstrated by an ELISA type assay of PGE2 in the supernatant of the cell cultures.

  b) Results Table 4: Inhibition of PGE2 following inflammation induced by PMA Stress PMA% inhibition of PGE2 Positive control (PMA alone) 0 Negative control (indomethacin + PMA) 86 Albizia adianthifolia leaf extract at 95 2.10-3% (w / v) Table 5: Inhibition of PGE2 following UV induced inflammation UV stress% inhibition of PGE2 Positive control (PMA alone) 0 Negative control (indomethacin + PMA) 86 Albizia adianthifolia leaf extract at 55 3% (w / v) Note: 100% PGE2 release is set for the positive control which corresponds to 0% inhibition.

  Inflammation induced by the chemical agent (PMA) or UV stress is very strongly inhibited by the extract of Albizia adianthifolia leaves.

  The extract of Albizia adianthifolia leaves therefore has anti-inflammatory activity.

EXAMPLE 4

  Inhibition of Melanogenesis - Bleaching A) Enzymatic Tests: Anti-Tyrosinase Activity This extract was tested according to the modified method of Hamada and Mishima (British Journal of Dermatology, 86, 385, 1972).

  In this technique, dopamine is contacted with tyrosinase in buffered medium at pH 7 and converted to dopachrome. Dopachrome is a colored molecule that absorbs radiation at 475 nm. The formation of the dopachrome can thus be followed in time at 475 nm. The activity of tyrosinase is translated by the speed of formation of dopachrome. The concentration of extract sufficient to decrease the rate of 50% (IC 50) is determined.

  The Cl 50 of Albizia adianthifolia leaf extract is 0.212 mg / ml. This extract therefore has significant anti-tyrosinase activity compared to kojic acid (1 mg / ml) considered as the reference.

  B) Cell Tests: Inhibition of Melanin Synthesis a) Materials and Methods The test is carried out on transformed melanocytes (S91) known to synthesize a great deal of melanin. The cells are treated for 72 hours with Albizia adianthifolia leaf extract at concentrations 0.001 and 0.002% after seeding, the treatment being renewed daily for 4 days. In parallel, a negative control with untreated cells and a positive control using kojic acid are made.

  After establishing a correlation of the mitochondrial activity as a function of the number of cells, the melanins produced by the cells are assayed by a spectrophotometric method.

  b) Results Table 6: inhibition of melanin synthesis% production melanins% inhibition melanins Control 100.00 0 Ac. kojic 1mM 65.41 34.59 Test extract at 69.22 30 '78 0.002% Test extract at 77.36 22 '63 0.001% 2871691 11 The negative control is fixed at 100% synthesis of melanins and therefore at 0 % inhibition. The 1 mM kojic acid makes it possible to inhibit melanin synthesis by 34.95%.

  An inhibition of melanin synthesis is obtained for the two concentrations tested for Albizia adianthifolia leaf extract. This inhibition is dose-dependent since the synthesis of melanins is more inhibited when the cells are treated with 0.002% of extract (30.78% inhibition), than with 0.001% of extract (22.63% inhibition). ) of melanin synthesis.

  The extract of Albizia adianthifolia leaves therefore has an inhibitory action on melanin synthesis.

EXAMPLE 5: Slimming

  A) Enzymatic Tests: Inhibition of Phosphodiesterase Albizia adianthifolia leaf extract was tested according to the modified method of Nicholson and Jackman (Brit J. Pharmacol., 97 (3), 889-897, 1989).

  In this technique, tritiated cyclic AMP is contacted with phosphodiesterase 1 and a conformational change can be observed, which is detectable by quantification of radioactivity. This transformation depends on the activity of the enzyme is monitored according to the concentration of the extract.

  The results of these experiments show that Albizia adianthifolia leaf extract inhibits up to 79% of the activity of phosphodiesterase 1 and therefore has an inhibitory activity of this enzyme, thus a lipolytic activity.

  B) Cell Tests: Determination of Triglycerides a) Materials and Methods From cells isolated from human adipose tissue, cell differentiation is induced by insulin, dexamethasone, indomethacin and 3-Isobutyl-1-methylxanthine . This differentiation causes the formation of lipid droplets and the accumulation of intracellular triglycerides visible in optical microscopy. To characterize this transformation, a quantification of the lipid droplets is made using the Nile Red dye. This test is performed on human preadipocytes in culture. After three days of culture, differentiation is induced in the presence of Albizia adianthifolia leaf extract solubilized in DMSO (dimethyl sulfoxide).

  On D9, D14 and D16, a triglyceride assay is performed using the AdipoRedTM reagent (Cambrex, PT-7009). The reading is done by fluorescence reader and the triglyceride level is calculated by the following ratio (RFU: fluorescent unit ratio): RFU at 4851590nm (test extract) Triglyceride level = RFU at 4851590nm (negative control) Then the rate of Adipocyte conversion is calculated (versus inhibition rate) knowing that the positive control is set at 100% conversion.

  b) Results Table 7: Inhibition of adipocyte conversion Inhibition rate of adipocyte conversion (in%) at day 9 to day 14 to day 16 Negative control 93 99 100 proliferation medium Positive control 0 0 0 differentiation medium Inhibitory reference 95 99 98 Naiquebaine at 20NM Test extract at 2.10-3% (w / v) 40 39 (action of deduced DMSO) 36 The leaf extract of Albizia adianthifolia thus has an inhibitory activity of adipocyte conversion and consequently a lipolytic activity.

  Example 6: Formulations Example of composition in emulsion form (% by weight of the composition). Water

  Albizia dry extract: Blend of preservatives: Propylene glycol: Xanthan gum: Acrylic / acrylate copolymer: Stearic acid 100 EO: Sorbitan stearate: Sorbitan laurate 20 EO: Cetylstearic alcohol: Beeswax: Wheat germ oil: Dimethicone : Cyclomethicone: Polyacrylamide gel: Fragrance:

QSP

  0.01% to 0.5% 0/0 1.50% 5.00% 0.30% 0.50% 3.00% 2.00% 3.00% 1.50% 1.00% 5.00 % 2.00% 5.00% 2.00% 0.10% Example of composition in the form of cream (% by weight of the composition). Water

  Albizia dry extract: Xanthan gum: Sequestering agent: Preservatives: C18 acid: C 16 acid: Trilaurine: Shea butter: Tocopherol acetate: Beta bisabolol Wheat germ oil Dimethicone: Polyacrylic acid: TEA (triethanolamine): Fragrance:

QSP

  0.01% to 0.5% 0.30% 0.05% 1.50% 2.50% 2.50% 1.00% 0.05% 0.05 0.05% 5.00% 3, 00% 0.30% 1.50% 0.10

Claims (12)

CLAIMS:   1. Cosmetic composition comprising an extract of Albizia adanthifolia leaves.   2. Cosmetic composition according to claim 1, wherein said extract is obtained by hydroalcoholic extraction.   3. Cosmetic composition according to any one of the preceding claims, wherein the extract is obtained by extraction into an ethanol / water mixture in proportions 70/30 by volume.   4. Cosmetic composition according to any one of the preceding claims, intended for topical use.   5. Cosmetic composition according to claim 4, which is in the form of a single emulsion oil in water or water in oil, multiple emulsion, microemulsion, aqueous or aqueous-alcoholic gel, cream, oil, aqueous lotion or hydroalcoholic.   6. Use of a cosmetic composition as defined in one of claims 1 to 5, for combating skin aging.   7. Use of a cosmetic composition as defined in one of claims 1 to 5, for protecting the skin from external aggression.   8. Use of a cosmetic composition according to claims 6 or 7, to fight against photoaging.   9. Use of a cosmetic composition as defined in one of claims 1 to 4 for the bleaching and / or depigmentation of the skin.   10. Use of a cosmetic composition as defined in one of claims 1 to 4 as a slimming cosmetic composition.   11. Use of a leaf extract of Albizia adanthifolia for the preparation of a pharmaceutical or cosmetic composition useful for the prevention or treatment of cutaneous inflammations.   12. Use of a leaf extract of Albizia adanthifolia for the preparation of a pharmaceutical or cosmetic composition for the prevention or treatment of cellulite.