USE OF AN EXTRACT OF LYTHRUM SALICARIA

Abstract

The present invention relates to the cosmetic and/or nutraceutical use of an extract of the Lythrum salicaria plant and/or of gallic acid and/or one of the gallic acid derivatives, in particular vescalagin and/or castalagin, more preferentially castalagin, for maintaining and/or increasing collagen expression in the skin and/or the mucous membranes for maintaining and/or increasing the firmness of the skin and/or the mucous membranes and/or for maintaining and/or increasing lipolysis in skin adipocytes for inducing a slimming effect onto the skin and/or for decreasing, preventing and/or treating the unaesthetic manifestations of cellulite, for example the orange peel appearance of the skin and/or the jodhpur thigh appearance.

Another subject of the invention relates to a cosmetic care process comprising the application, preferably topically, of the Lythrum salicaria extract according to the invention and/or of gallic acid and/or of one of the gallic acid derivatives, in particular vescalagin and/or castalagin, more preferentially castalagin, or of a cosmetic composition comprising it, for maintaining and/or increasing collagen expression in the skin and/or the mucous membranes for increasing the firmness of the skin and/or of the mucous membranes and/or maintaining and/or increasing lipolysis in skin adipocytes for inducing a slimming effect onto the skin and/or decreasing, preventing and/or treating the unaesthetic manifestations of cellulite, for example the orange peel appearance of the skin and/or the jodhpur thigh appearance.

Claims

1-26. (canceled)

27. A cosmetic method for maintaining and/or increasing firmness of skin and/or mucous membranes, comprising administering to a human being whose skin and/or mucous membranes is in need thereof an effective amount of a Lythrum salicaria extract and/or gallic acid and/or one of the gallic acid derivatives.

28. The method of claim 27, wherein the Lythrum salicaria extract and/or gallic acid and/or one of the gallic acid derivatives maintains and/or increases collagen expression in the skin and/or the mucous membranes.

29. The method of claim 27, for inducing a slimming effect onto the skin and/or decreasing, preventing and/or treating unaesthetic manifestations of cellulite.

30. The method of claim 29, wherein the Lythrum salicaria extract and/or gallic acid and/or one of the gallic acid derivatives maintains and/or increases lipolysis in skin adipocytes.

31. The method of claim 27, wherein the Lythrum salicaria extract is an extract obtained by aqueous extraction.

32. The method of claim 27, wherein the Lythrum salicaria extract is an extract of aerial parts of the plant.

33. The method of claim 28, wherein the collagen is type I collagen.

34. The method of claim 27, wherein the Lythrum salicaria extract is obtained by extraction of an amount of from 0.1% to 10% by weight of dry matter of at least one part of the Lythrum salicaria plant relative to the total weight of the solvent/plant mixture (w/w).

35. The method of claim 27, wherein the administration consists in the topical application to at least one area the skin and/or the mucous membranes in need thereof of the Lythrum salicaria extract and/or gallic acid and/or one of the gallic acid derivatives.

36. The method of claim 27, wherein the Lythrum salicaria extract and/or gallic acid and/or one of the gallic acid derivatives is solubilized in an aqueous solution comprising hexylene glycol and caprylyl glycol or a mixture thereof.

37. The method of claim 27, wherein the Lythrum salicaria extract comprises at least 0.25% by weight of vescalagin and/or castalagin relative to the total weight of the extract.

38. The method of claim 27, wherein the Lythrum salicaria extract comprises at least 20% by weight of maltodextrin relative to the total weight of the extract and the maltodextrin.

39. The method of claim 27, wherein the Lythrum salicaria extract and/or gallic acid and/or one of the gallic acid derivatives is present in a cosmetic composition comprising at least one cosmetically acceptable excipient.

40. The method of claim 39, wherein the Lythrum salicaria extract is present in the composition at a concentration of from 1×10.sup.4% to 10% by weight relative to the total weight of the composition.

41. The method of claim 39, wherein the gallic acid and/or one of the gallic acid derivatives is present in the composition at a total final concentration of molecules of from 2.5×10.sup.−7 to 0.1% relative to the total weight of the composition.

42. The method of claim 39, wherein the cosmetic composition is in form of an aqueous or oily solution, a cream or a gel which is aqueous or an oily gel, a shampoo, a milk, an emulsion, a microemulsion or a nanoemulsion, a mask, a serum, a lotion, a liquid soap, a dermatological bar, an ointment, a foam, or a patch.

43. The method of claim 27, wherein the Lythrum salicaria extract and/or gallic acid and/or one of the gallic acid derivatives is present in a nutraceutical composition, and wherein the administration is an oral administration of the nutraceutical composition being in form of gel capsules, capsule, oral powder, gel and/or oral liquid.

44. The method of claim 35, wherein the area of skin and/or mucous membrane is chosen from the stomach, the arms, the forearms, the knees, the bust, the thighs, the hips, the buttocks, the neck, the waist and/or the face.

45. A method for the treatment and/or prevention of telangiectasia, comprising administering to a human being in need thereof of an effective amount of a Lythrum salicaria extract and/or gallic acid and/or gallic acid derivative or a dermatological composition comprising it with a dermatologically acceptable excipient.

46. A method for the prevention and/or treatment of cellulite, comprising administering to a human being in need thereof of an effective amount of a Lythrum salicaria extract and/or gallic acid and/or gallic acid derivative or a dermatological composition comprising it with a dermatologically acceptable excipient.

47. The method of claim 46, wherein the extract is an extract of Lythrum salicaria obtained by aqueous extraction.

Description

EXAMPLE 1: VARIOUS EMBODIMENTS OF THE INVENTION FOR OBTAINING A LYTHRUM SALICARIA EXTRACT ACCORDING TO THE INVENTION

[0119] a) Obtaining an extract of the aerial parts of Lythrum salicaria according to the Invention

[0120] Five percent of aerial parts, that is to say a mixture of leaves, flowers, flower peduncles and stem, by weight of dry matter relative to the total weight of the mixture consisting of water and of the aerial parts of the Lythrum salicaria plant (w/w), were macerated in water, the water being in this case the sole solvent, for a period of 2 hours at ambient temperature, that is to say at 20° C. After maceration, the crude extract obtained was centrifuged for 10 min (8000 rpm) and then the supernatant was recovered and filtered (cut-off threshold of 0.45 μm). The extract of aerial parts at 5% (w/w) was then tested with regard to the increase in type I collagen protein expression under the conditions presented in the example 2 hereinafter.

b) Obtaining an Extract of the Aerial Parts of Lythrum salicaria According to the Invention

[0121] Five percent of aerial parts, that is to say of a mixture of leaves, flowers, flower peduncles and stem, by weight of dry matter relative to the total weight of the mixture consisting of water and of the aerial parts of the Lythrum salicaria plant (w/w), were macerated in water at a temperature of 80° C. for a period of 1 hour. The crude extract was then centrifuged for 10 min (8000 rpm) and then the supernatant was filtered (0.45 μm).

c) Obtaining an Extract of the Aerial Parts of Lythrum salicaria According to the Invention

[0122] Five percent of aerial parts, that is to say of a mixture of leaves, flowers, flower peduncles and stem, by weight of dry matter relative to the total weight of the mixture consisting of water and of the aerial parts of the Lythrum salicaria plant (w/w), were macerated at a temperature of 4° C., for a period of 16 hours, in water. The crude extract was then centrifuged for 10 min (8000 rpm) and then the supernatant was filtered (0.45 μm).

d) Obtaining an Extract of the Aerial Parts of Lythrum salicaria According to the Invention

[0123] One percent of aerial parts by weight of dry matter relative to the total weight of the mixture consisting of water and of the aerial parts of the Lythrum salicaria plant (w/w) were macerated at a temperature of 4° C. for a period of 16 hours in water. The crude extract was then centrifuged for 10 min (8000 rpm) and then the supernatant was filtered (0.45 μm).

e) Obtaining an Extract of Lythrum salicaria Leaves According to the Invention

[0124] Five percent of leaves by weight of dry matter relative to the total weight of the mixture consisting of the solvent and of the leaves (w/w) of the Lythrum salicaria plant were macerated in water as sole solvent for a period of 1 hour at a temperature of 80° C. The crude extract was then centrifuged for 10 min (8000 rpm) and then the supernatant was filtered (0.45 μm).

f) Obtaining an Extract of Lythrum salicaria Leaves According to the Invention

[0125] An amount of 5% of Lythrum salicaria leaves by weight of dry matter relative to the total weight of the mixture consisting of water and of the Lythrum salicaria plant was macerated at ambient temperature, in this case 20° C., for a period of 2 hours, in a water/butylene glycol mixture (75/25; w/w). The crude extract was then centrifuged for 10 min (8000 rpm) in order to remove the insoluble fraction and then the supernatant was filtered (0.45 μm).

q) Obtaining a Lythrum salicaria Extract Comprising Vescalagin and/or Castalagin According to the Invention

[0126] An amount of 10% of the aerial parts, that is to say of the mixture of leaves, flowers, flower peduncles and stem of Lythrum salicaria, by weight of dry matter relative to the total weight of the mixture consisting of the solvent and of the aerial parts of the plant, was macerated with stirring at ambient temperature, in this case 20° C., for a period of 24 hours, in a water/ethanol mixture (75/25; w/w).

[0127] The extract was then purified in a first step by solid phase extraction (SPE) on a C18-grafted silica solid phase. The extract was deposited in aqueous-alcoholic solution containing 20% of methanol and 0.5% of formic acid. The elution was carried out with this same solvent. The extract obtained was thus concentrated by evaporation and then lyophilized. The extract obtained was then purified in a second step by semi-preparative high performance liquid chromatography (HPLC) on a Synergi Fusion (Phenomenex) column having the dimensions 250×10 mm. The elution was carried out with the gradient of 0.5% formic acid and methanol.

[0128] The purified extract thus obtained comprises vescalagin and/or castalagin, in a total amount of molecules of 1% by weight relative to the total weight of the purified extract.

[0129] Maltodextrin was added to the extract and then the extract was concentrated by evaporation and then lyophilized.

EXAMPLE 2: DEMONSTRATION OF THE EFFECT OF A LYTHRUM SALICARIA EXTRACT ACCORDING TO THE INVENTION ON TYPE I COLLAGEN PROTEIN EXPRESSION

Protocol:

[0130] The experimental conditions for demonstrating the increase in type I collagen protein expression are those set out in example 1 of the international application published under number WO 2012/175454.

[0131] Normal, that is to say non-pathological, human fibroblasts obtained from abdominal biopsies from a healthy donor were seeded into 96-well plates and cultured in a defined medium (FGM) up to 100% confluence, which is obtained after 3 days of culture.

[0132] The Lythrum salicaria extract according to the invention, prepared according to example 1 a of the present invention, was added at a final concentration of 1% (w/w) to the culture medium. The same culture medium without the addition of extract according to the invention was used as a control (non-treated control).

[0133] After 48 hours of culture post-confluence at 37° C., the culture medium was removed and eliminated. A cell lysis step was carried out and the lysate was then removed and analysed. A DNA assay was carried out on the lysates so as to express the collagen protein expression per cell.

[0134] The double-stranded DNA was assayed by the bisbenzimide method (Invitrogen, Quant-iT™ PicoGreen® dsDNA). The DNA concentration is proportional to the number of viable cells and makes it possible to rationalize the fluorescence read with respect to a number of cells.

[0135] The type I collagen was assayed by means of the immunochemical technique as follows:

[0136] An anti-type I collagen antibody was incubated for 30 minutes with the cell lysate. After rinsing with PBS, the europium-coupled secondary antibody (Perkin Elmer) was added. A revealing solution was added and the fluorescence was measured using an EnVision multiplate reader (Perkin Elmer).

[0137] For each condition, the time-resolved fluorescence (TRF) was measured in each well and rationalized with respect to the amount of DNA assayed in the well. The (fluorescence/DNA concentration) ratio was calculated.

[0138] For each condition, the results are expressed by the percentage of the mean of the protein expressions relative to the protein expressions measured in the control (non-treated control) (n=6).

Results:

[0139]

TABLE-US-00001 Standard Mean deviation Control 100 1 1% (w/w) Lythrum salicaria extract 1a) 196.6 14.5

Conclusion:

[0140] The Lythrum salicaria extract according to the invention induced an increase in the type I collagen protein expression in human fibroblasts cultured in vitro. The extract according to the invention can thus be used for increasing the firmness of the skin and/or the mucous membranes.

EXAMPLE 3: EFFECT OF VARIOUS LYTHRUM SALICARIA EXTRACTS ACCORDING TO THE INVENTION ON THE INCREASE IN LIPOLYSIS OF SKIN ADIPOCYTES

Example 3a) Lythrum Salicaria Extract According to Example 1a)

Protocol:

[0141] Adipocytes were obtained by differentiation of human preadipocytes as follows: normal, that is to say non-pathological, non-cancerous, human preadipocytes were inoculated into a growth medium enriched with growth factors (PAGS 7252 Cliniscience) and foetal calf serum (FBS). After 3 days of culture, the culture medium was replaced with a differentiation medium (PADM 7221 Cliniscience) containing the growth factors and the FBS. After 2 days of culture, the differentiation medium was replaced with a standard medium containing 1% of FBS. The adipocytes formed were then incubated for a period of 16 hours in the presence of the Lythrum salicaria extract according to example 1a) at a final concentration of 1% by weight relative to the total weight of the final culture medium and of the extract.

[0142] The lipolysis was evaluated by quantification of the glycerol released into the culture medium. The quantification of the glycerol was obtained by means of an enzymatic method (Libios K-GCROL kit) and measurement of the optical density at 340 nm. The results are expressed as percentage relative to the control (culture medium without L. salicaria extract) (Base 100). The results presented are the mean of 3 assays (n=3).

Results:

[0143]

TABLE-US-00002 Standard Mean deviation Control 100 20.2 1% (w/w) Lythrum salicaria extract 1a) 210.3 31.8

Conclusion:

[0144] The results show an increase in lipolysis in the adipocytes cultured in the presence of the Lythrum salicaria extract, showing the capacity of the extract to induce a slimming effect and to decrease the unaesthetic manifestations of cellulite, for example the orange peel appearance and/or the jodhpur thigh appearance.

Example 3b) Lythrum salicaria Extract Comprising Vescalagin and Castalagin According to Example 1g)

Protocol:

[0145] Adipocytes were obtained by differentiation of 3T3-L1 murine preadipocytes as follows: the murine preadipocytes were inoculated into a growth medium enriched with neonatal calf serum. After 3 days of culture, the culture medium was replaced with a differentiation medium containing the differentiation factors Iso-Butyl-Methyl-Xanthine (IBMX) and Dexamethasone, and foetal calf serum (FCS). After 2 days of culture, the differentiation medium was replaced with a standard medium containing 10% of FCS and incubated for 8 days. The adipocytes formed were then incubated for a period of 16 hours in the presence of the Lythrum salicaria extract according to example 1g) at a final concentration of 2.5×10.sup.−3% by weight relative to the total weight of the final culture medium and of the extract.

[0146] The lipolysis was evaluated by quantification of the glycerol released into the culture medium. The quantification of the glycerol was obtained by means of an enzymatic method (Libios K-GCROL kit) and measurement of the optical density at 340 nm.

[0147] The results are expressed as percentage relative to the control (culture medium without L. salicaria extract) (Base 100). The results presented are the mean of an assay carried out in “quadruplet” (n=4).

Results:

[0148]

TABLE-US-00003 Standard Mean deviation Control 100 12 2.5 × 10.sup.−3% (w/w) Lythrum salicaria extract 1g) 151 4

Conclusion:

[0149] The results showed an increase of at least 35% in lipolysis in the murine adipocytes cultured in the presence of the Lythrum salicaria extract comprising vescalagin and castalagin, showing the capacity of the extract to induce a slimming effect and to decrease the unaesthetic manifestations of cellulite, for example the orange peel appearance and/or the jodhpur thigh appearance.

EXAMPLE 4: EXAMPLE OF COSMETIC AND/OR DERMATOLOGICAL INGREDIENTS CONTAINING THE LYTHRUM SALICARIA EXTRACT ACCORDING THE INVENTION

[0150] The methods known to those skilled in the art were carried out in order to mix together the various ingredients according to the present invention. The percentages expressed are percentages by weight relative to the total weight of the composition.

a) The Lythrum salicaria Extract According to the Invention is the One Obtained According to Example 1a

TABLE-US-00004 Lythrum salicaria extract (Ex. 1a))   5% Hexylene glycol 0.5% Caprylyl glycol   1% Xanthan gum 0.5% Water qs 100
b) The Lythrum salicaria Extract According to the Invention is the One Obtained According to Example 1b

TABLE-US-00005 Lythrum salicaria extract (Ex. 1b))   5% Hexylene glycol 0.5% Caprylyl glycol   1% Xanthan gum 0.5% Water qs 100
c) The Lythrum salicaria Extract According to the Invention is the One Obtained According to Example 1c

TABLE-US-00006 Lythrum salicaria extract (Ex. 1 g) ) 1-5% Maltodextrin  80% Water qs 100

EXAMPLE 5: EXAMPLE OF COSMETIC COMPOSITIONS CONTAINING THE EXTRACT ACCORDING TO THE INVENTION

[0151] The compositions hereinafter are prepared according to methods known to those skilled in the art, in particular as regards the various phases to be mixed together.

5a) Cosmetic Composition Containing a Lythrum salicaria Extract According to the Invention

TABLE-US-00007 Cosmetic ingredient* 3.00 EDTA 0.05 Steareth-2 2.00 Steareth-21 2.50 Cetearyl alcohol 1.00 Propylheptyl caprylate 15.00 Sodium hydroxide (30% in solution) 0.10 Mixture of phenoxyethanol, chlorphenesin, benzoic acid, 1.25 butylene glycol, sorbic acid (Germazide ™ PBS) Mixture of polyacrylate-X, isohexadecane and 4.00 polysorbate 60 (Sepigel ™ SMS 60) Water qs 100 *The cosmetic ingredient is prepared according to example 4a or 4c above. The amounts indicated are in percentage by weight relative to the total weight of the composition.
5b) Cosmetic Composition Comprising a Lythrum salicaria Extract According to the Invention

TABLE-US-00008 Cosmetic ingredient* 3.00 EDTA 0.05 Steareth-2 2.00 Steareth-21 2.50 Cetearyl alcohol 1.00 Propylheptyl caprylate 15.00 Sodium hydroxide (30% in solution) 0.10 Mixture of phenoxyethanol, chlorphenesin, benzoic acid, 1.25 butylene glycol, sorbic acid (Germazide ™ PBS) Mixture of polyacrylate-X, isohexadecane and 4.00 polysorbate 60 (Sepigel ™ SMS 60) Water qs 100 *The cosmetic ingredient is in this case prepared according to example 4b or 4c above. The amounts indicated are in percentage by weight relative to the total weight of the composition.

EXAMPLE 6: EXAMPLE OF A DERMATOLOGICAL COMPOSITION IN OINTMENT FORM CONTAINING THE EXTRACT ACCORDING TO THE INVENTION

[0152] The composition hereinafter is prepared according to methods known to those skilled in the art, in particular as regards the various phases to be mixed together.

[0153] The amounts indicated are in percentage by weight relative to the total weight of the composition.

TABLE-US-00009 Ingredient* 3.00 Excipient: Low-density polyethylene 5.50 Liquid paraffin qs 100 *The ingredient is prepared according to example 4b or 4c above. The ingredient according to the invention is in this case sterilized and then dried before being incorporated into the composition in ointment form.