AU2017345384B2 - Extract of Anigozanthos flavidus for cosmetic use - Google Patents

Abstract

The invention relates to the use of an extract of

Description

EXTRACT OF ANIGOZANTHOS FLAVIDUS FOR COSMETIC USE

Field of the invention The present invention relates to the cosmetic field, in particular to anti-ageing cosmetic agents.

Context of the invention The skin is the first barrier of protecting the organism against outside attacks. This organ is composed of several layers of tissue. The epidermis which is the outermost part of the skin, the dermis, a connective tissue consisting of fibroblasts and an extracellular matrix, which performs the skin cohesion and nutrition functions, and the hypodermis consisting of adipocytes, are singled out. The epidermis consists of several cell strata of keratinocytes. The germ layer of the epidermis, called the basal layer, containing in particular the skin stem cells, the spiny layer, Stratum spinosum, consisting of several layers of polygonal cells, the granular layer, Stratum granulosum, comprising one to three layers of flattened cells containing cytoplasmic inclusions, the keratohyalin granules, and, finally, the horny layer, Stratum corneum which is composed of anucleate cells rich in keratin, called corneocytes, which correspond to the terminal keratinocyte differentiation stage, are inter alia, singled out. The outermost cells of the horny layer are continually eliminated and replaced with the cells of a lower layer, according to a process known as desquamation. The cell regeneration of the horny layer is based on a process of cell maturation in which the cells of the basal layer of the epidermis differentiate and gradually migrate through the various strata of the epidermis until they reach the horny layer in the form of corneocytes. Skin ageing, whether it results from a normal phenomenon of senescence or is accentuated by an external factor such as exposure to UV radiation, involves dysfunctions of cell differentiation and/or renewal resulting in atrophy of all the layers of the skin. From a histological point of view, a decrease in the quality of the dermis, in particular a decrease in the thickness of the epidermis and a loss of consistency of the extracellular matrix, are, inter alia, observed. It is thus observed, in the extracellular matrix, that the collagen fibers become less numerous and less regular and that the ratio of collagen type III to collagen type I is increased. The number of elastic fibers of the papillary dermis is also decreased and the elastic fibers of the reticular dermis are altered. From an esthetic point of view, these alterations result in a modification of the appearance of the skin and of its mechanical properties: the skin is less smooth, rougher, its microrelief is more marked, and it can exhibit fine lines, which may with time result in the formation of deep wrinkles. The skin also exhibits a loss of elasticity, of tone and of firmness, that can result in tissue sagging or slackening and an accentuation of the wrinkles, in particular on the face or the neck. Skin ageing can have an impact on the uniformity of the complexion and can lead to the appearance of pigment spots. There are numerous anti-ageing cosmetic products on the market, intended to prevent or attenuate slackening of the skin. Nevertheless, there remains, at the current time, a need for new anti-ageing active agents for preventing or treating alterations resulting from skin ageing, in particular linked to skin slackening.

Summary of the invention The invention relates to the cosmetic use of an extract of Anigozanthosfavidus as an anti ageing agent. In particular, the extract of Anigozanthos flavidus can be used as an anti-wrinkle, smoothing, toning, restructuring or remodeling agent for the skin. Typically, the extract of Anigozanthos flavidus is present as an active agent in a cosmetic composition intended for preventing, delaying and/or treating one or more signs of skin ageing. The sign(s) of skin ageing can be selected from the group consisting of: the appearance of fine lines and/or wrinkles on the skin, in particular at the lips or on the eye lids, withering or sagging of the skin, in particular on the neck, a loss of density of the skin, a loss of firmness of the skin, a loss of tonicity of the skin, a loss of the elasticity of the skin, a loss of contractility of the skin, a loss of maintenance of the skin, a slacking of the skin, an alteration of the smooth appearance of the skin, an increase in the roughness of the skin, and an alteration of the outline of the face. The extract of Anigozanthos flavidus can be obtained from an aerial part of Anigozanthus flavidus, for example the flowers, the leaves, the stems, and combinations thereof. In certain embodiments, the extract of Anigozanthos flavidus is obtained by extraction with a polar solvent, preferably under hot conditions.

As mentioned above, the extract of Anigozanthosflavidus may be present as an active agent in a cosmetic composition. Said cosmetic composition may comprise at least one additional cosmetic agent, preferably selected from the group consisting of vitamins, sun filters and sunscreens, anti-ageing or antiwrinkle agents, antioxidants, lifting agents, firming agents, anti-spot agents, anti-redness agents, slimming agents, draining agents, moisturizing agents, soothing agents, scrubbing or exfoliating agents, matting agents, sebum regulators, lightening active agents, self-tanning active agents, tanning accelerators and combinations thereof. In certain embodiments, the cosmetic composition is selected from the group consisting of aqueous solutions, aqueous-alcoholic solutions, oil-in-water (O/W) or water-in-oil (W/O) or multiple (triple: W//W or O/W/O) emulsions, nanoemulsions, in particular O/W nanoemulsions, the drop size of which is less than 100 nm, aqueous gels or dispersions of a fatty phase in an aqueous phase by means of spherules, suspensions, preferably in aqueous or aqueous-alcoholic media, liposome suspensions, powders, lotions, milks, creams, salves, gels, foams and ointments. The cosmetic composition may be selected from a cosmetic product, a makeup product or a body hygiene product, for example a lotion, a milk, a serum, an aqueous or oily gel, an emulsion, a cream, in particular a day cream or a night cream, a cream-gel, a skincare water, an ointment, a balm, a foundation, a spray, an eyeshadow, a stick, a lipstick, a gloss, a lip balm, a foam, a deodorant, a nourishing mask, a shower gel, and an exfoliating or scrubbing product. The invention also relates to a cosmetic composition for use in the prevention or treatment of a sign of ageing, said composition comprising an extract of Anigozanthos flavidus, in particular as defined above, and one or more pharmaceutically acceptable excipients. According to an additional aspect, the invention relates to a "precursor" composition, that is to say a composition intended for the preparation of a cosmetic composition comprising: - from 10% to 50%, preferably from 20% to 40% of an extract of Anigozanthos flavidus, preferably a liquid extract of Anigozanthosflavidus, - from 50% to 90%, preferably from 60% to 80% of a carrier, preferably selected from an aqueous solvent, an organic solvent, preferably a lower alcohol such as ethanol, propanediol, butylene glycol, glycerol or isopropanol, a lipophilic agent and mixtures thereof, and - optionally, from 0.1 to 30% of a cosmetically acceptable additional excipient, preferably selected from a pH adjuster, an antioxidant, a preservative, a stabilizer, a vectorizing agent, a thickener, an emulsifier, a hydrophilic or lipophilic gelling agent, a fragrance, a mineral or organic oil, and combinations thereof, the percentages being expressed by weight relative to the total weight of said composition. By way of example, said composition for the preparation of a cosmetic composition comprises: - from 20% to 40 % of an aqueous extract of Anigozanthosflavidus, and - from 60% to 80% of glycerol, the percentages by weight being expressed relative to the total weight of said composition. An object of the invention is also a process for preparing a cosmetic composition according to the invention, comprising a step of mixing a "precursor" composition according to the invention with one or more acceptable excipients from a cosmetic point of view and/or with one or more additional active agents having a cosmetic effect.

Presentation of the figures Figure 1 shows the amount of procollagen I expressed as a percentage relative to the Control produced by fibroblasts incubated with TGF-P or with a 0.1% concentration of the composition referred to as "Extract" (Example 1). The "Extract" composition comprises 5% by weight of an aqueous extract of Anigozanthosflavidus flowers, less than 35% by weight of water, and more than 50% by weight of glycerol. Figure 2 shows the amount of Tenascin-X expressed as a percentage relative to the Control produced by fibroblasts incubated with fetal calf serum (FCS), with a 0.2% concentration of the "Extract" composition (Extract 0.2%) and in the presence of a 0.1% concentration of the "Extract" composition (Extract 0.1%) (Example 2). Figures 3A-C present the results relating to the effect of the "Extract" on the contractile forces of the human fibroblasts from the bottom of a wrinkle (BW) (Example 4). Figure 3A shows the change in the contractile forces applied by the BW fibroblasts in the presence of 0.1% (v/v) of the "Extract" composition, in the presence of 0.2% (v/v) of the "Extract" composition, and in the absence of active agent (Control (-)), for 24h. The histograms of Figure 3B correspond to the areas under the curve (AUC) of the curves of the Figure 3A. The histograms of Figure 3C correspond to the maximum contraction forces for the curves of Figure 3A.

BW: BW fibroblast without active agent (negative control), BW fibroblasts incubated in the presence of 0.1% (v/v) of the "Extract" composition (Active agent 0.1%), and of 0.2% (v/v) of the "Extract" composition (Active agent 0.2%) Figure 4 shows the images of immunolabeling of a-SM actin fibers in fibroblasts from the bottom of a wrinkle: controls (A), after incubation with 0.2% of "Extract" (B) and after incubation with 0.1% of "Extract" (C). Figures 5A and 5B show the contractile forces exerted by the non-cellular equivalent dermis as a function of time in the presence of the "Extract" composition (Active agent at 0.2%) (Kangaroo Paw 0.2%) and in the absence of said composition (Control). Figure 6 shows the variation in the depth of the crow's feet wrinkles of the face, treated with a placebo cream (cream A) in comparison with a cream containing the "Extract" at 2% (cream B), lh after the application of the Extract and after 28 days of treatment.

Description of the invention Anigozanthos flavidus is an endemic plant species of the family Haemodoraceae that is encountered mainly in south-west Australia. This plant is characterized by a flower, which is generally yellow or red in color, and the shape of which is reminiscent of a kangaroo paw. To the knowledge of the applicant, the extracts of Anigozanthosfiavidus have been used in cosmetics, at the very most, as a conditioning agent, for example, in shampoo compositions. No cosmetic biological activity seems to have been described for the extracts of Anigozanthosflavidus. The applicant has prepared an aqueous extract of Anigozanthosfavidus and has studied the biological activities of this extract. Surprisingly, the applicant has shown that the extract of Anigozanthos flavidus is active on biological targets which play a key role in the organization and composition of the extracellular matrix of the skin. As is illustrated in the examples, the extract of Anigozanthos flavidus is capable of stimulating the synthesis of procollagen I by the dermal fibroblasts and of increasing the synthesis of collagen XVII by the keratinocytes. The applicant has also shown that the extract of Anigozanthos flavidus is capable of stimulating the synthesis of tenascin-X, a glycoprotein which plays a key role in the structure and stability of the elastic fibers and in the organization of the collagen fibers of the extracellular matrix of the skin. The extract of Anigozanthos flavidus also stimulates the synthesis of actin alpha 2 (also known as alpha smooth muscle actin (a-SMA)) in fibroblasts, which results also in an increase in the contractile forces developed by the fibroblasts from the bottom of wrinkles. It has also been shown that the extract of Anigozanthosfavidus is capable of increasing the contractile forces exerted by a non-cellular equivalent dermis, which suggests an immediate tensioning effect of the extract on the fibers of the dermis via a physical action. The applicant has also carried out clinical tests aimed at evaluating the anti-ageing effects of a day cream comprising 0.6% by weight of an aqueous extract of Anigozanthosflavidus. These trials have shown that the extract of Anigozanthosflavidus exerts a tensioning remodeling effect on the skin by significantly reducing the depth of the wrinkles in the long term. It has also been shown that the extract of Anigozanthosflavidus exerts an immediate tensioning effect, that can be observed from the first minutes after application thereof to the skin. With regard to these results, the extracts of Anigozanthosfavidus therefore have applications in the cosmetics field as anti-ageing agents for preventing or treating the signs of skin ageing. The extracts of Anigozanthosfavidus also have applications as immediate-action tensioning agents in order to re-tighten the skin and/or to improve the appearance or the maintenance of the skin, of the skin, in the short term. By virtue of the actions on the synthesis of several key proteins involved in the organization, cohesion and elasticity of the extracellular matrix of the skin, the extracts of Anigozanthos flavidus can be used to promote the restructuring and reorganization of the extracellular matrix, in particular of the dermis in mature or elderly skin. In general, the extracts of Anigozanthosfavidus can be used for treating or preventing skin slackening, improving skin elasticity, or else treating or preventing wrinkles and fine lines of the skin. Thus, the extracts of Anigozanthos flavidus can be used as tensioning or lifting agents or as agents for remodeling and restructuring of the skin. In particular, the extracts of Anigozanthosfavidus can be used as a cosmetic agent for treating or preventing a non-pathological alteration of the skin linked to a loss of elasticity of the skin and/or to an alteration of the extracellular matrix of the skin, for example a disorganization of the extracellular matrix resulting from a qualitative or quantitative alteration of the constituents of the extracellular matrix, such as the elastic fibers.

Uses of the extract of Anigozanthos flavidus accordingto the invention

According to a first aspect, the present invention relates to the non-therapeutic, cosmetic use of an extract of Anigozanthosfiavidus. The extract of Anigozanthosfiavidus can be used as a cosmetic agent, in particular as an anti-ageing agent. In the context of the present invention, an extract of Anigozanthosfavidus refers to an extract obtained from an aerial part of the Anigozanthos flavidus plant, such as the leaves, the flowers, the stems and combinations thereof. In one particular embodiment, the extract of Anigozanthos flavidus is an extract obtained from the flowers and/or the stems of Anigozanthos favidus. The extract of Anigozanthos favidus can be obtained by any known extraction technique. Preferably, the extract of Anigozanthos flavidus is prepared by extraction with a polar solvent. Typically, the polar solvent can be selected from water, an alcohol, an ester, a ketone and mixtures thereof. By way of example of esters, mention may be made of ethyl acetate, propyl acetate and butyl acetate. The alcohols encompass, without being limited thereto, methanol, ethanol, n-propanol, isopropanol, methanol, and polyols such as glycerol. By way of example of ketones, mention may be made of acetone, cyclohexanone or else methyl ethyl ketone. In one particular embodiment, the extraction solvent is selected from water, an alcohol and an aqueous-alcoholic mixture, for example a water/ethanol, water/methanol or water/glycerol mixture. In certain particular embodiments, the extraction solvent is water. The extraction with the polar solvent can be carried out by any method known to those skilled in the art, for example by decoction, infusion, maceration, percolation or leaching. For example the extract of Anigozanthosfavidus can be obtained by hot extraction in water or in an aqueous-alcoholic mixture. A suitable hot extraction technique is decoction. The term "decoction" is intended to mean boiling a plant material in water for a period of time suitable for obtaining an efficient extraction. The extraction step can be carried out on a plant material of Anigozanthosfiavidus that has undergone one or more treatments such as freezing, drying, freeze-drying, milling or combinations thereof. In certain embodiments, the extraction is carried out on a fresh or dried plant material having undergone a milling step. At the end of the extraction, the extract can be recovered by any method which makes it possible to separate the plant material from the liquid phase, for example by centrifugation and/or filtration. By way of example, the extract of Anigozanthos flavidus can be obtained by a process comprising:

- a step of preparing a plant material by drying and milling an aerial part, typically the flowers and the stems, of Anigozanthosfiavidus, - a step of extracting the plant material with a polar solvent, typically water or an aqueous-alcoholic mixture, preferably under hot conditions, for example by decoction, and - a step of eliminating the solid plant material, preferably by filtration. The process for preparing the extract of Anigozanthosflavidus may comprise one or more additional steps at the end of the extraction and/or filtration step. The process may comprise a step of formulating the extract, for example by adding a pH adjuster, a stabilizer, and/or a cosmetically acceptable carrier or support, a step of concentrating the extract, or else a step of drying the extract. In one additional or alternative embodiment, the extract of Anigozanthosfavidus comprises flavonoids, polyphenols and carbohydrates. The flavonoids can represent from 1% to 15%, for example from 5% to 15% or 1% to 8% in naringin equivalence relative to the dry matter of the extract; the polyphenols can represent from 5% to 30%, for example from 5% to 25%, from 12% to 20%, from 10% to 30%, or from 15% to 20%, in chlorogenic acid equivalence by weight relative to the dry matter of the extract; and the sugars can represent from 35% to 65%, for example from 35% to 55%, from 40% to 60% or from 40% to 50%, in glucose equivalence by weight relative to the dry matter of the extract. The polyphenol, flavonoid and sugar contents are determined by colorimetry using a standard range obtained from chlorogenic acid (method of Folin-Ciocalteu), naringenin (method of Zhishen et al., Food Chemistry, 1999, 64, 555-559) and glucose (method of Bachelier et al., Cah. OSTRUM.s6r. P6dol. Vo. IV,3-1966) respectively. The extract of Anigozanthos flavidus obtained at the end of the extraction process can comprise a dry matter content ranging from 0.1% to 40% by weight, preferably from 0.1% to 10% by weight, for example from 0.1% to 4% by weight. In certain embodiments, the extract of Anigozanthos flavidus comprises less than 1% by weight of ferulic acid relative to the dry matter of the extract. Preferably, the extract of Anigozanthosfavidus is devoid of ferulic acid. In one particular embodiment, the extract of Anigozanthosflavidus is characterized by one or more (e.g. 2, 3, 4, 5 or all) of the following features:

- The extract of Anigozanthosfiavidus is obtained from an aerial part of Anigozanthos flavidus by extraction with a polar solvent, preferably by decoction in water or in an aqueous-alcoholic mixture, - The extract of Anigozanthosfiavidus comprises a dry matter content of from 0.1% to 10% by weight, - The flavonoids represent from 5% to 15%, for example from 7% to 13%, in naringin equivalence by weight relative to the dry matter of the extract, - The sugars represent from de 35% to 65%, for example from 40% to 60%, in glucose equivalence by weight relative to the dry matter of the extract, - The polyphenols represent from 5% to 25%, for example from 10% to 20%, in chlorogenic acid equivalence by weight relative to the dry matter of the extract, and/or - The extract of Anigozanthos flavidus comprises less than 1% by weight of ferulic acid relative to the dry matter of the extract. In the uses according to the invention, the extract of Anigozanthosfiavidus may be in liquid form, for example in water or in an aqueous-alcoholic mixture such as a water/glycerol mixture. In other embodiments, the extract of Anigozanthos flavidus may be in dry solid form, typically in the form of a powder. A dry extract of Anigozanthosflavidus can be obtained by eliminating the extraction solvent by any known technique, for example by spray-drying, evaporation or freeze-drying. By way of additional example, the extract of Anigozanthos flavidus may be in a form absorbed onto a solid carrier such as maltodextrin. In certain embodiments, the extract of Anigozanthos flavidus is a liquid extract in water or in an aqueous-alcoholic mixture, preferably in a water/glycerol mixture. Said aqueous alcoholic mixture may represent a water/alcohol volume ratio ranging from 1/9 to 9/1, preferably from 1/9 to 2/1. For the purposes of the invention, the term "anti-ageing agent" is intended to mean a cosmetic active agent which has a biological activity making it possible to prevent or treat a sign of skin ageing. The expression "preventinga sign ofskin ageing" is intended to mean delaying or preventing the appearance of the sign of skin ageing. The expression "treating a sign of skin ageing" is intended to mean decreasing, reducing, softening, correcting or slowing down the development of the sign of skin ageing.

The expression "a sign of skin ageing" is intended to mean any non-pathological alteration or modification of the visual appearance or of the mechanical properties of the skin, resulting from ageing, whether it is chronological (chronoageing) and/or photo-induced (photo ageing). A sign of skin ageing thus refers to a non-pathological alteration or modification of the appearance or of the mechanical properties of the skin. Thus, the signs of ageing encompass, without being limited thereto, thinning of the skin, in particular of the epidermis, the appearance of a microrelief, the appearance of fine lines and/or wrinkles on the skin, including at the lips and on the eyelids, withering or sagging of the skin, a loss of radiance of the skin, a dull complexion, pigment spots on the skin, dark circles under the eyes, a loss of density of the skin, a loss of firmness of the skin, a loss of tonicity of the skin, a loss of contractility of the skin, a loss of elasticity of the skin, an alteration of the smooth appearance of the skin, an increase in the roughness of the skin and an alteration of the outline of the face. For the purpose of the invention, the terms "skin" and "cutaneous" refer to any part of the skin of the human body, in particular the skin of the face, including the lips and the eyelids, the neck, and the skin of the hands. As presented above, the extract of Anigozanthos flavidus is capable of promoting the synthesis of various key proteins involved in the structure of the extracellular matrix and in the elasticity of the skin. Thus, the extract of Anigozanthos flavidus can be used to improve, restore, or prevent the alteration of the mechanical properties of the skin, in particular the suppleness, the elasticity, the firmness or the tonicity of the skin. For example, the extract of Anigozanthosflavidus can be used to improve, restore, or prevent the alteration of the mechanical properties of mature skin or of elderly skin. The term "mature skin" is intended to mean the skin of a man or a woman over the age of 40 and exhibiting a loss of elasticity due to ageing. In one particular embodiment, the extract according to the invention is used to treat or prevent a sign of the skin ageing associated with an alteration of the mechanical properties of the skin, in particular a loss of elasticity of the skin. The extract of Anigozanthosflavidus can thus be used as an agent for smoothing, increasing, lifting or relifting, restructuring or remodeling the skin. The extract of Anigozanthosflavidus can also be used as an agent for immediate tensioning of the skin.

The expression "an agentfor smoothing" is intended to mean a cosmetic agent capable of reducing and/or correcting the microrelief of the skin, including of the lips, said microrelief being in the form of wrinkles or fine lines. The expression "agentforlifting or relifting" is intended to mean a cosmetic agent capable of exerting a tensioning action on the skin, by virtue of which the appearance of the skin is improved and the wrinkles are less visible. The expression "agent for remodeling the skin" is intended to mean an agent capable of reducing the irregular appearance of the skin subsequent to a loss of elasticity or a slackening of the skin, in particular at the level of the outline of the face and the neck. The term "lifting agent" is intended to mean a cosmetic agent capable of exerting a tensioning effect on the skin so as to soften the microrelief of the skin. The term "restructuringagent" is intended to mean a cosmetic agent capable of preventing or treating an alteration of the extracellular matrix of the skin resulting from ageing, such as a loss or an alteration of the elastic fibers of the extracellular matrix or else a loss or an alteration of the collagen of the extracellular matrix. The expression "immediate tensioning agent or immediate-effect tensioning agent" is intended to mean a cosmetic agent capable of exerting a tensioning effect on the skin so as to increase the elasticity and/or improve the appearance or the maintenance of the skin, it being possible for this effect to be visible typically between a few minutes (for example five minutes) and one hour after application to the skin. By way of example, the extract of Anigozanthosflavidus can be used in order to prevent or to treat the appearance of fine lines and/or wrinkles on the skin, in particular at the lips or on the eyelids, withering of the skin, for example on the neck, sagging or slackening of the skin, a loss of density of the skin, a loss of firmness of the skin, a loss of tonicity of the skin, a loss of elasticity of the skin, a slackening of the skin, an alteration of the smooth appearance of the skin, an increase in the roughness of the skin, and an alteration of the outline of the skin. The extract of Anigozanthosflavidus can also be used to give mature or elderly skin a more youthful appearance and/or to improve the complexion or the radiance of mature or elderly skin. The extract of Anigozanthosflavidus can be used for cosmetic purposes for restructuring or reorganizing the extracellular matrix of skin impaired because of ageing. In particular, the extract of Anigozanthos flavidus can be used for cosmetic purposes for stimulating the synthesis of a protein involved in the organization of the extracellular matrix of the skin. The expression "protein involved in the organization of the extracellular matrix of the skin" is intended to mean a constituent protein of the extracellular matrix, such as the collagens, elastin or fibrillin, or a protein involved in the expression or in the organization of the constituent proteins of the extracellular matrix, such as tenascin-X. In certain embodiments, the extract of Anigozanthosflavidus is used as a cosmetic agent for stimulating the synthesis of a protein selected from procollagen I, collagen XVII, and tenascin-X in the skin, preferably in mature skin. In another embodiment, the extract of Anigozanthos flavidus is used as a cosmetic agent for increasing the contractile forces of the dermal fibroblasts of mature or elderly skin. In another embodiment, the extract of Anigozanthosflavidus is used to stimulate dermal metabolism. In certain embodiments, the extract of Anigozanthosflavidus is used in combination with an active agent having a cosmetic effect. Such agents are described below and encompass, inter alia, antiwrinkle agents, anti-ageing agents, antioxidants, moisturizing agents, lifting agents, and firming agents.

In the uses according to the invention, the extract of Anigozanthosflavidus is applied to the skin. Said extract is preferably present as a cosmetic agent in a composition intended to be applied to the skin. Said composition is typically a cosmetic composition. The extract of Anigozanthosflavidus generally represents from0.0001% to 10% by weight, preferably from 0.001% to 5% by weight, more preferably from 0.005% to 3% by weight, relative to the total weight of said cosmetic composition. By way of example, the extract of Anigozanthos flavidus may represent from 0.01% to 3,0% by weight, for example 0.6% by weight, of the cosmetic composition. The extract of Anigozanthosflavidus may be added to the cosmetic composition in the form of a liquid extract, in the form of a dry extract or else in the form of a "precursor" composition as described below in the present description. Said cosmetic composition may comprise one or more cosmetically acceptable excipients and optionally one or more additional active agents having a cosmetic effect. Typically, said cosmetic composition comprises: - from 0.0001% to 10% of extract of Anigozanthosflavidus, - from 0% to 20% of one or more additional active agents, and - from 70% to 99.9999% of one or more cosmetically acceptable excipients, the percentages being expressed by weight relative to the total weight of the cosmetic composition.

In certain embodiments, said cosmetic composition comprises - from 0.001% to 5% by weight of extract ofAnigozanthosflavidus, - from 0.001% to 10% of one or more additional active agents, and - from 85% to 99.998% of one or more cosmetically acceptable excipients. In other embodiments, said cosmetic composition comprises: - from 0.01% to 5% by weight of extract of Anigozanthosflavidus, - from 0.01% to 10% of one or more additional active agents, and - from 85% to 99.98 % of one or more cosmetically acceptable excipients. The extract of Anigozanthosflavidus is preferably present in liquid form. In one additional or alternative embodiment, the cosmetic composition comprises from 0.001% to 10%, for example from 0.005% to 1% or from 0.005% to 0.5% by weight of dry matter from the extract of Anigozanthosflavidus. The expression"active ingredienthaving a cosmetic effect", "active agent having a cosmetic effect" or "cosmetic or active agent having a cosmetic effect" is intended to mean a compound capable of exerting at least one cosmetic effect on the skin or skin appendages. The term "cosmetic effect" is intended to mean any non-therapeutic effect aimed at modifying and/or improving the appearance of the skin or of the mucous membranes, such as the lips, at protecting them against outside attacks (sun, wind, moisture, dryness, chemical products), or else at preventing and/or at correcting phenomena associated with ageing thereof. Thus, in certain embodiments, the extract of Anigozanthos flavidus may be present in a cosmetic composition which also comprises an additional active ingredient having a cosmetic effect. This active ingredient having a cosmetic effect may be selected from the group consisting of vitamins, sun filters and sunscreens, anti-ageing agents, anti-redness agents, antioxidants, firming agents, moisturizing agents, soothing agents, scrubbing or exfoliating agents, matting agents, sebum regulators, lightening active agents, anti-spot active agents, slimming agents, draining agents, self-tanning active agents, tanning accelerators and combinations thereof. In particular the cosmetic composition may comprise tocopherols, and/or extracts of plants, such as extracts of linseeds, extracts of Vibrio exopolysaccharides, or peptides such as trifluoroacetyl tripeptide-2. Preferably, the cosmetic composition may comprise an active agent selected from an antiwrinkle agent, an anti-redness agent, an antioxidant, a moisturizing active agent, a soothing agent, a sebum regulator, an anti-spot agent, a sun filter or sunscreen, and combinations thereof. More preferably, the additional active agent having a cosmetic effect is selected from an antiwrinkle agent, an antioxidant, a moisturizing agent, a firming agent and combinations thereof. By way of example of moisturizing agents, mention may be made of urea, pidolic acid (PCA) and derivatives thereof, in particular salts thereof such as arginin PCA, chitosan PCA, copper salts (copper PCA), magnesium salts (magnesium PCA), sodium salts (sodium PCA) or zinc salts thereof, ethylhexyl PCA, calcium gluconate, hyaluronic acid and salts thereof and other glycosaminoglycans, fructose, glucose, isomaltose, lactose, trehalose, polydextrose, saccharose (sucrose), maltitol, mannitol, sorbitol, xylitol and other carbohydrates and derivatives, polyethylene glycols such as PEG-7, PEG-8, PEG-10, PEG-12 or PEG-14, glycerol, propylene glycol, butylene glycol, betaine, citrulline, collagen and derivatives thereof, histidine, silk hydrolysates, keratin hydrolysates or soya hydrolysates, plant extracts rich in polysaccharides and/or polyphenols, for example extracts of Aloes, cornflower (Centaurea cyanus), and combinations thereof. By way of examples of anti-ageing agents, mention may be made of ascorbic acid and derivatives thereof, such as magnesium ascorbyl phosphate, glycosaminoglycans and derivatives thereof, Cyathea polysaccharides, collagen, linseed (Linum usitatissimum) extracts, peptides such as Caprooyl Tetrapeptide-3 and trifluoroacetyl tripeptide-2, extracts of Polygonum aviculare, extracts of brown algae, in particular of Ascophyllum nodosum, extracts of ferns, in particular of Cyathea cumingii. By way of example of soothing agents, mention may be made of allantoin, extracts of aloes, of birch (for example Betula alba), of willowherb (Epilobium angustifolium), of chestnut (for example Castenea sativa), of cornflower (for example Centaurea cyanus), of centella (for example Centella asiatica),of field horsetail (for example Equisetum arvense), of fennel (for example Foeniculum vulgare), of common witch hazel (for example Hamamelis virginiana),of ivy (for example Hedera helix), of Habiscus sabdariffa, of lily (for example Lilium candidum), of common mallow (for example Malva sylvestris), of lemon balm (for example Melissa officinalis), of skullcap (for example Scutellaria baicalensis), of mimosa (for example Mimosa tenuiflora),of cinquefoil (for example Potentilla erecta), an extract of oligosaccharides or an oligosaccharide, for example of flax, peptides such as palmitoyl tripeptide-8, and combinations thereof. By way of example of antioxidants, mention may be made of HMR (hydroxymethyl resorcinol), ascorbic acid and derivatives thereof, vitamin B9, histidine hydrochloride, or an extract of willowherb (Epilobium augustifolium). The active ingredients having an antioxidant effect and of vitamin type are generally used in a weight percentage of at least 1% relative to the total weight of the cosmetic composition. By way of example of sebum regulators, mention may be made of flax lignans, rice powder, zinc gluconate, sarcosine, an extract of Cinnamomum zeylanicum bark, an extract of avocado, and combinations thereof. By way of anti-redness agents, mention may be made of saponins, flavonoids, ruscogenins, esculosides, and extracts containing them, for example extracts of Ruscus, and also certain essential oils, for example of lavender or of rosemary. By way of example of anti-spot agents, mention may be made of extracts such as licorice (Glycyrrhiza glabra), extract of jackfruit (Artocarpus heterophyllus), extract of western dock (R.occidentalis),extracts of a plant belonging to the Citrus genus, resveratrol, peptides such as oligopeptide-68, nonapeptide-1, kojic acid, magnesium ascorbyl phosphate and combinations thereof. The cosmetically acceptable excipient(s) present in the cosmetic composition may be selected from diluents, dispersants, gelling agents, emollients, vectorizing agents such as polycationic polymers or phospholipids, gums, resins, solvents, in particular lower alcohols, especially ethanol, isopropanol, dipropylene glycol, butylene glycol, propanediol, glycerol, sorbitol, and propylene glycol, fillers such as modified and polymerized starches, titanium dioxide, or a metal stearate, preservatives, essential oils, pearlescent agents, dyes, odor absorbers, pH regulators or neutralizing agents, lubricants, thickeners, surfactants, including anionic, cationic, amphoteric or nonionic surfactants, humectants, wetting agents, fragrances, organic or else mineral pigments, such as iron oxides, oily agents such as oils or fats of plant origin, fats of animal origin, synthetic oils such as petroleum jelly, silicone oils (cyclomethicone), fatty alcohol esters, fluoro oils, waxes, modified clays, bentones, metal salts of fatty acids, silica, polyethylenes, mica, preservatives, antimicrobial agents, carriers such as a mineral, spring or floral water, and/or other substances commonly used in formulation in the cosmetics or pharmaceutical field. The extract of Anigozanthos flavidus can be incorporated into any type of cosmetic composition. Preferably, it is a cosmetic composition having a form suitable for topical administration, in particular suitable for application to the skin. Said cosmetic composition may be in the form of aqueous or aqueous-alcoholic solutions, of oil-in-water (O/W) or water-in-oil (W/O) or multiple (triple: W/O/W or O/W/O) emulsions, nanoemulsions, in particular O/W nanoemulsions, the drop size of which is less than 100 nm, of aqueous gels, of dispersions or else of a powder. The cosmetic composition according to the invention may be in the form of a lotion, a milk, a cream, a salve, a gel, a foam, a solution or an ointment. More generally, the composition according to the invention may also be in the form of a cosmetic product of any type. It may be a cosmetic care product or a makeup or body hygiene product, for example a lotion, a milk, a serum, an aqueous or oily gel, an emulsion, a cream, a cream-gel, a skincare water, an ointment, a balm, a foundation, a spray, an eyeshadow, a stick, a lipstick, a gloss, a foam, a deodorant, a nourishing mask, a shower gel, or an exfoliating or scrubbing product. By way of nonlimiting illustration, the extract of Anigozanthos flavidus may be present as an anti-ageing, relifting or remodeling agent in a day cream intended for mature or elderly skin. The extract of Anigozanthosflavidus may be present as an anti-ageing agent in a regenerating serum intended for the face or the neck. By way of additional example, the extract of Anigozanthos flavidus may be present as a smoothing or remodeling active agent in a care cream for the hands, the neck or the chest.

In certain embodiments, the extract of Anigozanthos flavidus is present in the cosmetic composition in combination with a vectorizing agent. In certain embodiments, the extract of Anigozanthos flavidus is formulated by means of a vectorizing system. The vectorizing system may be selected from the group consisting of micelles, liposomes, including unilamellar or multilamellar liposomes, niosomes, ethosomes, lamellar systems, nanosomes, lipid or polymeric vesicules, nanospheres, micro- or nanoparticles of polymers which may or may not be natural, hydrogels. Preferably, the extract of Anigozanthos flavidus is encapsulated in a vectorizing system selected from liposomes and lamellar systems. Particular examples of liposomes and lamellar systems for implementing the present invention are described, inter alia, in French patent applications No. 1358589 and No. 1262303.

An additional object according to the invention is a cosmetic process for treating or preventing a sign of skin ageing in a subject, comprising the administration of a cosmetically effective amount of the extract of Anigozanthosfiavidus, preferably by topical route, to said subject. The extract of Anigozanthos flavidus is typically applied to the skin to be treated, for example on the face, the neck or the hands. Preferably, the extract of Anigozanthos flavidus is present in a cosmetic composition as described above, and is therefore applied in a form of a cosmetic composition, for example in the form of a cream. As mentioned above, the sign of skin ageing can result from an alteration of the mechanical properties of the skin, in particular a loss of elasticity of the skin. Preferably, the cosmetic process according to the invention has the objective of treating or preventing a sign of skin ageing selected from the appearance of wrinkles and/or fine lines, of pigment spots on the skin, withering of the skin, sagging or slackening of the skin, a loss of density of the skin, a loss of firmness of the skin, a loss of tonicity of the skin, a loss of elasticity of the skin, slackening of the skin, an alteration of the smooth appearance of the skin, an increase in the roughness of the skin, an alteration of the outline of the face, and combinations thereof. An object of the invention is also a cosmetic process for exerting an immediate tensioning effect on the skin, preferably on mature skin, comprising the administration of a cosmetically effective amount of the extract of Anigozanthosfavidus, preferably topically, to said subject. The extract of Anigozanthosfavidus is typically applied to the skin to be treated, for example on the face, the neck or the hands. Another object according to the invention is a cosmetic process for reorganizing or restructuring the extracellular matrix of the skin and/or for stimulating the synthesis of a protein selected from procollagen I, collagen XVII, and tenascin-X in the skin, preferably in mature skin, and/or for increasing the contractile forces of the dermal fibroblasts of mature skin, said process comprising the administration of a cosmetically effective amount of the extract of Anigozanthos flavidus, preferably topically, to said subject. In another embodiment, the extract of Anigozanthosfiavidusis used to stimulate dermal metabolism or to reorganize or restructure the dermis. In the cosmetic methods and uses according to the invention, the dose to be administered and the frequency of administration of the extract of Anigozanthosfavidus vary as a function of the desired cosmetic effect, of the characteristics of the subject, in particular of their sex, their age and their skin type. Typically, the extract of Anigozanthosfiavidus can be applied, to the area to be treated, once or twice a day, typically in the morning and/or the evening, for several consecutive weeks or even several months, for example at least for three months. The area to be treated is typically selected from the group consisting of the hands, the neck, the face or a part of the face such as the area around the eyes, the area around the lips and the region of the nasal grooves. The extract of Anigozanthos flavidus can be applied in the form of a cosmetic composition as described below. Said cosmetic composition typically comprises from 0.001% to 10%, for example from 0.01% to 5% by weight of extract.

By way of example, in order to obtain an anti-ageing effect on the face, the patient may apply a dose of 1 g to 2 g of cosmetic composition at 0.1% by weight of extract of Anigozanthos flavidus on their face, in the morning and evening. The subject may be a woman or a man, typically over the age of 30, preferably over the age of 40 or over the age of 45. For example, the subject may be a woman over the age of 45.

- Additional objects according to the invention According to an additional aspect, an object of the invention is also a cosmetic composition comprising an extract of Anigozanthos flavidus. Typically, said cosmetic composition comprises: - from 0.0001% to 10% of extract of Anigozanthosflavidus, - from 0% to 20% of one or more additional active agents, and - from 70% to 99.9999% of one or more cosmetically acceptable excipients, the percentages being expressed by weight relative to the total weight of the cosmetic composition. In certain embodiments, said cosmetic composition comprises - from 0.001% to 5% by weight of extract of Anigozanthosflavidus, - from 0.001% to 10% of one or more additional active agents, and - from 85% to 99.998% of one or more cosmetically acceptable excipients. In other embodiments, said cosmetic composition comprises: - from 0.01% to 5% by weight of extract of Anigozanthosflavidus, - from 0.01% to 10% of one or more additional active agents, and - from 85% to 99.98% of one or more cosmetically acceptable excipients. It goes without saying that the extract, the additional active agents and the pharmaceutically acceptable excipients are as described above. Likewise, the cosmetic composition may be of any type, as explained above. In particular, the extract of Anigozanthosfavidus may be in the form of a liquid extract. In one additional or alternative embodiment, the cosmetic composition comprises from 0.001% to 10%, for example from 0.005% to 1% by weight of dry matter from the extract of Anigozanthosflavidus.

According to an additional aspect, an object of the invention is a composition intended to be incorporated into a cosmetic composition. This composition thus corresponds to a "precursor"composition. Preferably, this composition for the preparation of a cosmetic composition comprises: - from 10% to 50%, preferably from 20% to 40% of an extract of Anigozanthos flavidus, - from 50% to 90%, preferably from 60% to 80% of a carrier, preferably selected from an aqueous solvent, an organic solvent, preferably a lower alcohol such as ethanol, propanediol, butylene glycol, glycerol or isopropanol, a lipophilic agent and mixtures thereof, and - optionally, from 0.01 to 30% of a cosmetically acceptable additional excipient, preferably selected from a pH adjuster, a buffer, a vectorizing agent, an antioxidant, a preservative, a stabilizer, a thickener, an emulsifier, a hydrophilic or lipophilic gelling agent, a fragrance, a mineral or organic oil, and combinations thereof, the percentages being expressed by weight relative to the total weight of the composition. It goes without saying that the extract of Anigozanthosfavidus may be as described above. By way of example, the extract of Anigozanthos flavidus may be an extract that can be obtained from an aerial part of Anigozanthosfiavidus by extraction with a polar solvent. In certain embodiments, the extract of Anigozanthosfavidus is a liquid extract, for example in water or in an aqueous-alcoholic mixture such as a water/glycerol mixture. In one particular embodiment, the composition for the preparation of a cosmetic composition comprises: - from 20% to 40 % of an aqueous extract of Anigozanthosfiavidus, and - from 60% to 80% of glycerol, the percentages being expressed by weight relative to the total weight of the composition. The "precursor" composition may also comprise a buffering agent or a pH adjuster such as citric acid. Typically, the "precursor" composition according to the invention has a pH of between 3.0 and 4.5, typically of approximately 3.9. In one additional or alternative embodiment, the "precursor" composition comprises from 0.01% to 10%, for example from 0.05% to 5% by weight or else from 0.1% to 1% by weight of dry matter from the extract of Anigozanthosfiavidus.

An object of the present invention is also the use of said "precursor" composition for the preparation of a cosmetic composition. It goes without saying that said final cosmetic or pharmaceutical composition is particularly suitable for the implementation of any one of the cosmetic uses according to the invention, and may have any one of the features described above. The "precursor" composition may be present in the cosmetic composition in an amount of from 0.001% to 50%, preferably from 0.01% to 10% by weight, for example from 0.5% to 5% or else from 1.5% to 2.5% by weight relative to the total weight of the final cosmetic composition or pharmaceutical composition. Said cosmetic composition may be obtained by mixing a "precursor" composition according to the invention with one or more excipients from a cosmetic point of view and/or with one or more additional active agents having a cosmetic effect.

An additional object according to the invention is a method for preparing a cosmetic composition according to the invention, comprising a step wherein an extract of Anigozanthos flavidus or the "precursor" composition is mixed with one or more cosmetically acceptable excipients and/or with one or more additional active agents having a cosmetic effect. An object of the invention is also a method for preparing a "precursor" composition according to the invention, comprising: a) The provision of a plant matrix of Anigozanthos flavidus, preferably from an aerial part of Anigozanthosflavidus, by drying and milling, b) The extraction of the plant matrix with a polar solvent, followed by a step of filtration so as to obtain a liquid extract, c) The addition of a cosmetically acceptable carrier to the liquid extract from the filtration, d) Optionally, the addition of a cosmetically acceptable excipient to the filtrate, e) Optionally, a pH adjustment step, It being possible for steps c), d) and e) to be carried out in any order. In certain embodiments, the cosmetically acceptable carrier comprises glycerol and/or water. The extraction step can be carried out as described above, for example using water or an aqueous-alcoholic mixture as polar solvent.

Other aspects and advantages of the present invention will emerge on reading the examples which follow, which should be considered to be illustraive and in no way limiting.

EXAMPLES The example which follow were carried out with a composition hereinafter referred to as "Extract", said "Extract" composition comprising: - 30% by weight of an aqueous extract of Anigozanthos flavidus flowers or stems containing approximately 1.6% by weight of solids, and - 70% by weight of glycerol. For the purposes of the invention, this is a "precursor" composition.

EXAMPLE 1: Effect of an extract of Anigozanthos flavidus on the synthesis of procollagen I by human dermal fibroblasts - Context

Collagens constitute a family of proteins highly involved in the mechanical properties of the skin. Procollagen I is the precursor of the collagen type I, which is a major constituent of connective tissue. With age, the collagen content in the skin decreases, which results in the formation of wrinkles or fine lines on the skin. The aim of this study was to determine the effects of an extract of Anigozanthosflavidus on the synthesis of procollagen by human dermal fibroblasts.

- Protocol

The "Extract" composition was tested at the following concentrations: 0.1% and 0.05% (v/v). TGF-j was chosen as positive control and tested at 10 ng/ml. The negative control

corresponds to cells not treated with the "Extract" composition or TGF. The fibroblasts were isolated from human dermis and maintained in a specific DMEM culture medium supplemented with 10% of fetal calf serum, 1% of antibiotics (penicillin/streptomycin) and 1% of L-glutamine, at 37C, under 5% of C02 and 95% humidity. 2.5 x 104 cells were seeded per microplate well, in a complete DMEM medium. The cells were placed in an incubator for 24h. The culture medium was then replaced with serum-free DMEM. The microplates were left for a further 24 hours in the incubator in order to reach cell quiescence. The "Extract" composition or the TGF-j was added at the desired concentration. The microplates were incubated for a further 24 hours. At the end of the incubation, the supernatants were removed and analyzed by ELISA (MK101Z, Ozyme) in order to quantify the procollagen I.

- Result

The results obtained are illustrated in figure 1. Figure 1 shows the percentage activation of procollagen I synthesis in the treated cells compared with the non-treated cells. The fibroblasts treated with the "Extract" composition or the TGFP produced a larger amount of procollagen I than the non-treated cells. The effect of the "Extract" composition on collagen I synthesis is dose-dependent. The extract of Anigozanthus flavidus therefore stimulates procollagen I in the dermal fibroblasts.

EXAMPLE 2: Effect of an extract of Anigozanthosflavidus on the synthesis of Tenascin X by human dermal fibroblasts Tenascin-X is a protein which regulates both the structure and the stability of the elastic fibers of the extracellular matrix (ECM). It also contributes to the organization of collagen fibers. Tenascin-X plays a key role in the rigidity and elasticity of connective tissues. A hereditary deficiency in tenascin-X can result in a genetic disease known as Ehlers-Danlos syndrome. The aim of this study was to determine the effect of the "Extract" composition on tenascin X synthesis by human dermal fibroblasts.

- Protocol

The "Extract" composition was tested at 0.2% and 0.1% (v/v). Fetal calf serum at 10% was used as positive control. The negative control corresponds to cells not treated with the Extract or with the fetal calf serum. The fibroblasts were isolated from human abdominal dermis and maintained in a specific DMEM culture medium supplemented with 10% of fetal calf serum, 1% of antibiotics (penicillin/streptomycin) and 1% of L-glutamine, at 37C, under 5% CO2 and 95% humidity. 2.5 x 104 cells were seeded per microplate well, in a complete DMEM medium. The cells were placed in an incubator for 24h. The culture medium was then replaced with serum-free DMEM. The microplates were left for a further 24 hours in the incubator in order to reach cell quiescence.

The "Extract" composition or the fetal calf serum was added at the desired concentration. After the addition, the cells were incubated for a further 24h. At the end of the incubation, the supernatants were removed and analyzed by ELISA in order to quantify the tenascin-X. In parallel, at the end of the incubation, the proteins contained in the cell lysates were quantified by the Bradford method.

- Results

The results obtained were subjected to a Student's t-test for a p-value < 0.05. The results are presented in figure 2. Figure 2 shows, for each experimental condition, the amount of tenascin-X present in the medium, expressed as a percentage relative to the total amount of proteins. It is noted that the "Extract" composition induces a dose-dependent increase in the amount of tenascin-X, compared with the negative control (non-treated cell). In other words, the extract of Anigozanthosflavidus stimulates tenascin-X in the human fibroblasts.

EXAMPLE 3: Effect of an extract of Anigozanthosflavidus on collagen XVII synthesis by human keratinocytes Collagen XVII is a transmembrane protein which plays an essential role in maintaining the bonding between intracellular and extracellular elements involved in epidermal adhesion. The aim of this study was to determine the effect of the "Extract" composition on collagen XVII synthesis by human keratinocytes.

- Protocol

The "Extract" composition was tested at a concentration of 0.4% (v/v). TGF-P was chosen as positive control and tested at 10 ng/ml. The negative control corresponds to cells not treated with the Extract or the TGF-P. The keratinocytes were isolated from a human epithelial cell line and cultured in DMEM supplemented with 10% of fetal calf serum, 1% of antibiotics (penicillin/ streptomycin) and 1% of L-glutamine, at 37C, under 5% C02 and 95% humidity. 5 x 104 cells were seeded per microplate well, in complete DMEM medium, and left in the incubator for 24h. The culture medium was then replaced with DMEM comprising 1% of fetal calf serum, 1% of antibiotics and 1% of L-glutamine. The microplates were left in the incubator for a further 24 hours.

At the end of the incubation, the "Extract" composition or the TGF- was added at the desired concentration. The microplates were incubated for a further 24 hours. The collagen XVII was detected and quantified by immunolabeling using an anti-collagen XVII antibody diluted to 1/100, and revealed with a secondary antibody coupled to fluorescein. The fluorescence intensity corresponds to the level of collagen XVII expression. The images were processed using the Image J software.

- Results

The fluorescence measurements were normalized relative to the measurement of the intensity of the fluorescence measured for the negative control. An increase of +27% in the intensity of the fluorescence for the cells incubated with the "Extract" composition was observed, thereby illustrating an increase in collagen XVII expression compared with the non-treated cells. The incubation with TGF-P (positive control) led to an increase of 257% of the fluorescence signal compared with the negative control. In summary, the extract of Anigozanthosflavidus stimulates collagen XVII synthesis by the human keratinocytes.

EXAMPLE 4: Evaluation of the tensioning effect of the Extract: quantification of the contraction forces generated by fibroblasts from the bottom of a wrinkle and quantification of the a-SM actin By means of the GlasBoxPlusTM system, it ispossible to quantify the tensioning effect of a cosmetic active agent on a dermal "equivalent" obtained from fibroblasts from the bottom of a wrinkle. The fibroblasts from the bottom of a wrinkle develop less contractile forces than those of the neighboring non-wrinkled skin. It is also possible to carry out immunolabeling of the a-SM actin in the dermal equivalents after the measurements of the contractile forces in the GlasBoxPlusTM system. The aim of this study was to show the effect of the Extract on fibroblast contractility and on the synthesis of a-SM actin, which is involved in cell plasticity, migration and motility.

• Protocol - Fibroblastculture Biopsies 2 mm in diameter were taken from a lift (surgical waste; 63-year-old woman) at the level of the non-wrinkled skin and within a wrinkle.

The conventional explantation technique is used for extracting the fibroblasts from the healthy skin (HF) and from the bottom of a wrinkle (BW). The explants are cultured in "Dulbecco's modified Eagle's medium" supplemented with 10% of fetal calf serum (FCS), 40 mg/l of gentamicin and 2 mg/l of fungizone (DMEMc) at 37C, in a humid atmosphere and in the presence of 5% C02. Gradually (over the course of about ten days), the fibroblasts migrate to the exterior of the explants. The culture medium is renewed twice a week. When the fibroblasts are in sufficient amount, they are detached under the action of trypsin-EDTA, subcultured and amplified under the same culture conditions. - GlasBoxPlusTM system and measurement of contractile forces The dermal equivalents develop in a culture dish that consists of eight rectangular cells. Placed into each one are two flexible slides, the lower parts of which consist of grids to which the dermal equivalent attaches when it polymerizes. The dermal equivalent develops between two slides so as to give a rectangular shape that is slightly shrunken at the center. This shape in conventional mechanics is referred to as a "diabolo" shape. Under the influence of the contraction forces developed by the fibroblasts, the slides distort. Their distortion is measured by means of optical fibers. This distortion is proportional to the force developed within the dermal equivalent. The measurements are carried out in real time using a PC acquisition card and suitable software. At confluence, the monolayer-cultured fibroblasts are trypsinized and then counted using a Mallassez slide in order to bring the cell suspension to 8 x 10 5 cells/ml. A dermal equivalent production medium is prepared by mixing six volumes of 1.76X medium (DMEMc, NaHCO3, NaOH, antibiotics, FCS), three volumes of rat tail collagen type I (2 mg/ml) and 1 volume of cell suspension (8 x 105 cells/ml). The mixture is poured into the rectangular cells of the GlasBoxPlusTM. A gel is formed in a few minutes at 37C. The various culture media containing the "Extract" composition (final concentration in the medium: 0.1% and 0.2% (V/V)), or the TGF 1 (2.5 ng/ml) (positive control) or devoid thereof (negative control) are added and the isometric forces are measured for 24 hours. - Labeling ofa-SMActin The immunolabeling of the a-SM actin fibers is carried out through the use of an anti-a-SM actin mouse monoclonal antibody. An anti-mouse second antibody coupled to FITC makes it possible to carry out and localize the immunolabeling. The nuclei were stained with

Hoechst dye. The observation was carried out by confocal microscopy at 488 nm and two photon excitation microscopy at 705 nm. Fragments of dermal equivalents are obtained using a biopsy punch 5 mm in diameter. The dermal equivalents are placed in cold acetone in order to permeabilize them. After washing with PBS, a solution of aqueous hydrogen peroxide is added in order to eliminate the endogenous peroxidases. After rinsing with PBS, the dermal equivalents are placed in a glycine solution in order to eliminate the background noise due to the aldehyde groups of the fixative. The dermal equivalents are then placed in a blocking solution which makes it possible to block the nonspecific binding that would be due to the secondary antibody. The tissues are placed in the primary antibody overnight. After washing with PBS, the secondary antibody is added. After one hour of incubation and washing with PBS, the nuclei are stained with Hoechst dye for 10 min. After rinsing in PBS, the dermal equivalents are mounted between slide and cover slip by means of Eukitt. The slides are observed under a confocal microscope (Zeiss LSM510 NLO) at x40 magnification and photographs are taken. - Statisticalanalyses For the study of the contractile forces, a two-factor analysis of variance is carried out, followed if necessary by a Fisher test. The significances are retained only when p<0.05. The areas under the curve (AUC) are calculated as is the slope (rate of contraction) and the maximum contraction, all of this by means of the GraphPad Prism@ 5 software. For these data, a one factor analysis of variance is carried out, followed if necessary by a Fisher test. The significances are retained only when p<0.05.

Results The contractile forces developed by the fibroblasts at the bottom of a wrinkle (BW) are significantly lower than those developed by the healthy fibroblasts (HF). It is observed that the AUCs (areas under the curve of the curve representing the contractile force as a function of time) and the maximum contraction are significantly reduced for the BW fibroblasts compared with the HF fibroblasts (data not shown). The incubation of the fibroblasts with TGF- 1induces a significant increase in the AUC and in the maximum contractile force, both in the BW fibroblasts and in the HF fibroblasts. The incubation with the "Extract" composition brings about a significant increase in the contractile forces in the BW fibroblast group: the maximum contractile force and the AUC are significantly increased. These results are illustrated by figures 3A, 3B and 3C. The "Extract" composition also increases overall the contractile forces in the HF fibroblasts, but the increase observed is smaller than that obtained for the BW fibroblasts. The immunolabeling experiments show that the "Extract" composition stimulates a-SM actin expression by the BW fibroblasts (cf. figure 4).

The extract of Anigozanthosflavidus significantly and specifically stimulates the contractile forces of the fibroblasts from wrinkled skin. It therefore exerts a tensioning and lifting effect on mature or elderly skin.

EXAMPLE 5: Evaluation of the immediate physical tensioning effect of the Extract. By means of the GlasBoxPlusTM system, it was also possible to quantify the immediate tensioning effect of the Extract on a non-cellular dermal equivalent, by measuring the contractile forces generated.

• Protocol - GlasBoxPlusTM system and measurement of contractileforces The non-cellular dermal equivalents polymerize in a culture dish which consists of eight rectangular cells. Placed into each one are two flexible slides, the lower parts of which consist of grids to which the non-cellular dermal equivalent attaches when it polymerizes. The polymerization of the collagen gel in the presence or absence of the Extract generates forces which cause distortion of the slides. This distortion is measured by means of optical fibers. It is proportionate of the force developed within the non-cellular dermal equivalent.

A dermal equivalent production medium was prepared by mixing 6 volumes of 1.76X medium (DMEMc, NaHCO3, NaOH, antibiotics, FCS), 3 volumes of rat tail collagen type I (2 mg/ml) and 1 volume of DMEMc. The mixture was poured into the rectangular cells of the GlasBoxPlusTM. A gel is formed in a few minutes at 37C. The various culture media containing the "Extract" composition (final concentration in the medium: 0.2% (V/V)) or devoid thereof (negative control) were added and the isometric forces were measured for 24 hours.

- Statisticalanalyses

For the study of the contractile forces, a two-way analysis of variance was carried out, followed if necessary by a Fisher test, p<0.05. The areas under the curve (AUC), the slope (contraction rate) and the maximum contraction were calculated using the GraphPad Prism@ 5 software. For these data, a one-way analysis of the variance was carried out, followed if necessary by a Fisher test (p<0.05).

• Results The contractile forces are significantly increased in the presence of the "Extract" at 0.2% compared with the Control group. This increase is significant 1h30 after the beginning of the measurement and extends at least up to the 11th hour in the presence of 0.2% of Extract (figures 5A and 5B). It is also observed that the AUC (area under the curve representing the contractile force as a function of time) and also the maximum contraction are significantly increased for the group treated with the Extract according to the invention compared with the control group, not treated with the Extract.

The extract of Anigozanthosflavidus significantly and specifically stimulates the contractile forces of the non-cellular dermal equivalents. The extract of Anigozanthos flavidus according to the invention therefore exerts an immediate, astringent, tensioning and lifting physical effect and can thus be used to exert an immediate tensioning effect on mature skin.

EXAMPLE 6: Examples of cosmetic compositions comprising the Extract - Anti-ageing cream (Cream B) comprising 2% of the "Extract"composition Phase INCI name % by weight A Water 89.40% Tetrasodium EDTA 0.10% Chlorophenesin 0.30% Phenoxyethanol 0.80% B Copolymer of sodium acrylates and lecithin 2.00% (Lecigel@) C Dicaprylyl carbonate 3.00% Sweet almond oil 2.00% Tocopherol and extract of sunflower 0.20% (Vitapherole@ E1000 D Fragrance 0.20% Glycerol and extract of Anigozanthos flavidus 2.00% ("Extract" composition)

The anti-ageing properties of cream B were evaluated in the clinical trial presented in example 7.

- Anti-ageing serum comprising 2% of the "Extract"composition

Phase INCI name % by weight A Water 83.67% Sodium phytate and water and alcohol (Dermofeel@PA-3) 0.15% Water and Cl 14700 0.28% Lysolecithin and Sclerotium gum and Xanthan gum and Pullulan (Ecogel TM) 2.00% B Isoamyl Laurate (Dermofeel@ Sensolv) 1.00% Shea butter (Lipex@ 102) 3.00% Sweet almond oil 3.00% Tocopherol and extract of sunflower 0.20% (Vitapherole@ E1000

C Glycerol and water and sodium levulinate and sodium anisate 4.00% Glycerol and extract of Anigozanthos flavidus 2.50% ("Extract" composition) D Fragrance 0.20%

- Anti-ageing balm comprising 2% of the "Extract"composition Phase INCI name % by weight A Water 76.90% Sodium phytate and water and alcohol (Dermofeel@PA-3) 0.10% Glycerol 5.00% Chlorphenesion 0.30% Phenoxyethanol 0.80% B Copolymer of sodium acrylates and lecithin (Lecigel@) 2.00% C Shea butter (Lipex@ 102) 5.00% Behenyl alcohol 2.00% Isononyl isononanoate 5.00% Glyceryl stearate citrate and polyglyceryl-3 stearate and hydrogenated lecithin (Heliofeel TM) 1.00%

Tocopherol and extract of sunflower 0.20% (Vitapherole@ E1000 D Glycerol and extract of Anigozanthos flavidus 1.50% ("Extract" composition) Fragrance 0.20%

EXAMPLE 7: Clinical evaluation of the antiwrinkle effect of the Extract. Creams tested: - Cream B (see example 5) comprising 2% of the "Extract" composition. - Cream A: Placebo cream having the same composition as cream B apart from the fact that the 2% of the "Extract" composition had been replaced with 2% of deionized water.

Protocol: A test for use of the cream A product (placebo cream) was carried out in comparison with the cream B product containing 2% of the "Extract" composition, according to the following protocol: 42 volunteers were included blind in the study and divided up into two groups according to a randomization list established by a dedicated program: - Group A receiving cream A: 21 women from 45 to 69 years old (average age: 56.4 years old), having all types of skin, of which 57% with sensitive skin, of phototypes II to IV. - Group B receiving cream B: 21 women from 46 to 77 years old (average age: 60.8 years old), having all types of skin, of which 48% with sensitive skin, of phototypes II to IV. The product was applied first time by the volunteers in the laboratory. The subsequent applications were carried out at the volunteers' homes, by self-application, at a rate of two applications a day. T+lh after the first application: evaluation of the tolerance of the product being trialed by assessment of the skin parameters by the dermatologist on an area of interest after removal of the product. T+7 days: Evaluation of the tolerance of the product being trailed by assessment of the skin parameters on the area of interest after 7 days of use (by telephone contact). T+28 days: Evaluation of the tolerance of the product being trialed, by the dermatologist according to the assessment of the skin parameters after 28 days of test. Self-evaluation, by the volunteers, of the qualities of the product being trailed, by means of a dedicated questionnaire. Moreover, an evaluation of the efficacy of the product with respect to the average depth of the crows' feet wrinkles was carried out. The efficacy measurements were carried out at TO, Tlh (i.e. one hour after the application) and T28 (i.e. after 28 days of application of the product tested). The measurement atlh made it possible to evaluate the immediate tensioning effect of the Extract. The measurement at T28 made it possible to evaluate the anti-ageing effect, in particular the antiwrinkle effect, in the long term, of the Extract. The evaluation of the wrinkle depth was carried out by quantifying the relief of the surface of the skin using the LifeViz MicroTM system (Quantificare). LifeViz MicroTM is a contactless method of measurement based on structured light projection on the skin areas of interest.

Results Cream B was well tolerated by the patients during the trial. The skins treated with cream B containing the extract of Anigozanthos flavidus showed a decrease in wrinkle depth of 10% which was significant, after 28 days of application, in comparison with the skins treated with cream A (placebo cream) (figure 6). A decrease in wrinkle depth of 5% is visible one hour after the application of cream B, underlining the immediate tensioning effect of the Extract according to the invention (figure 6).

The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates.

Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

Claims (25)

The claims defining the invention are as follows:

1. Use of an extract obtained from an aerial part of Anigozanthos flavidus in the manufacture of a cosmetic composition for treating skin ageing.

2. The use of the extract of Anigozanthos flavidus of claim 1, for treating wrinkles, smoothing, toning, lifting or relifting, restructuring or remodeling of the skin.

3. The use according to any one of the preceding claims, wherein the extract of Anigozanthosfavidus is present as an active agent in the cosmetic composition intended for preventing, delaying and/or treating one or more signs of skin ageing.

4. The use according to claim 3, wherein the sign(s) of skin ageing are selected from the group consisting of: the appearance of fine lines and/or wrinkles on the skin, in particular at the lips and on the eyelids, withering or sagging of the skin, in particular on the neck, a loss of radiance of the skin, a dull complexion, pigment spots, dark circles under the eyes, a loss of density of the skin, a loss of firmness of the skin, a loss of tonicity of the skin, a loss of elasticity of the skin, a loss of contractility of the skin, a loss of maintenance of the skin, slackening of the skin, an alteration of the smooth appearance of the skin, an increase in the roughness of the skin, and an alteration of the outline of the face.

5. The use according to any one of claims 1 to 4, wherein the extract of Anigozanthos flavidus is used for improving, restoring or preventing the alteration of mechanical properties of the skin in a mature skin.

6. The use according to any one of claims 1 to 4, wherein the extract of Anigozanthos flavidus is used for improving the complexion or the radiance of a mature skin.

7. The use according to any one of the preceding claims, wherein the extract of Anigozanthosfavidus is obtained from an aerial part of Anigozanthusfavidus selected from the group consisting of the flowers, the leaves, the stems and combinations thereof.

8. The use according to any one of the preceding claims, wherein the extract of Anigozanthosflavidus is obtained by extraction with a polar solvent.

9. The use according to any one of claims 1 to 8, wherein the cosmetic composition also comprises at least one additional cosmetic agent, preferably selected from the group consisting of vitamins, sun filters and sunscreens, anti-ageing or antiwrinkle agents, antioxidants, lifting agents, firming agents, anti-spot agents, anti-redness agents, slimming agents, draining agents, moisturizing agents, soothing agents, scrubbing or exfoliating agents, matting agents, sebum regulators, lightening active agents, self tanning active agents, bronzing accelerators and combinations thereof.

10. The use according to any one of claims 1 to 9, wherein the cosmetic composition is selected from the group consisting of aqueous solutions, aqueous-alcoholic solutions, oil-in-water (O/W) or water-in-oil (W/O) or multiple (triple: W/O/W or O/W/O) emulsions, nanoemulsions, in particular O/W nanoemulsions, with a drop size of less than 100 nm, aqueous gels, or dispersions of a fatty phase in an aqueous phase by means of spherules, suspensions, preferably in aqueous or aqueous-alcoholic media, liposome suspensions, powders, lotions, milks, creams, salves, gels, foams and ointments.

11. The use according to any one of claims 1 to 10, wherein the cosmetic composition is selected from a cosmetic product, a makeup product or a body hygiene product, for example a lotion, a milk, a serum, an aqueous or oily gel, an emulsion, a cream, in particular a day cream or a night cream, a cream-gel, a skincare water, an ointment, a balm, a foundation, a spray, an eyeshadow, a stick, a lipstick, a gloss, a lip balm, a foam, a deodorant, a nourishing mask, a shower gel, and an exfoliating or scrubbing product.

12. A method of treating skin ageing in a subject including the step of applying to the skin of said subject an effective amount of an extract obtained from an aerial part of Anigozanthosflavidus.

13. The method of claim 12, for treating wrinkles, smoothing, toning, lifting or relifting, restructuring or remodeling of the skin.

14. The method according to any one of claims 12 or 13, wherein the extract of Anigozanthos flavidus is present as an active agent in a cosmetic composition intended for preventing, delaying and/or treating one or more signs of skin ageing.

15. The method according to claim 14, wherein the sign(s) of skin ageing are selected from the group consisting of: the appearance of fine lines and/or wrinkles on the skin, in particular at the lips and on the eyelids, withering or sagging of the skin, in particular on the neck, a loss of radiance of the skin, a dull complexion, pigment spots, dark circles under the eyes, a loss of density of the skin, a loss of firmness of the skin, a loss of tonicity of the skin, a loss of elasticity of the skin, a loss of contractility of the skin, a loss of maintenance of the skin, slackening of the skin, an alteration of the smooth appearance of the skin, an increase in the roughness of the skin, and an alteration of the outline of the face.

16. The method according to any one of claims 12 to 15, wherein the extract of Anigozanthos flavidus is used for improving, restoring or preventing the alteration of mechanical properties of the skin in a mature skin.

17. The method according to any one of claims 12 to 16, wherein the extract of Anigozanthos flavidus is used for improving the complexion or the radiance of a mature skin.

18. The method according to any one of claims 12 to 17, wherein the extract of Anigozanthos flavidus is obtained from an aerial part of Anigozanthusflavidus selected from the group consisting of the flowers, the leaves, the stems and combinations thereof.

19. The method according to any one of claims 12 to 18, wherein the extract of Anigozanthos flavidus is obtained by extraction with a polar solvent.

20. The method according to any one of claims 14 to 19, wherein the cosmetic composition also comprises at least one additional cosmetic agent, preferably selected from the group consisting of vitamins, sun filters and sunscreens, anti-ageing or antiwrinkle agents, antioxidants, lifting agents, firming agents, anti-spot agents, anti-redness agents, slimming agents, draining agents, moisturizing agents, soothing agents, scrubbing or exfoliating agents, matting agents, sebum regulators, lightening active agents, self tanning active agents, bronzing accelerators and combinations thereof.

21. The method according to any one of claims 14 to 20, wherein the cosmetic composition is selected from the group consisting of aqueous solutions, aqueous-alcoholic solutions, oil-in-water (O/W) or water-in-oil (W/O) or multiple (triple: W/O/W or O/W/O) emulsions, nanoemulsions, in particular O/W nanoemulsions, with a drop size of less than 100 nm, aqueous gels, or dispersions of a fatty phase in an aqueous phase by means of spherules, suspensions, preferably in aqueous or aqueous-alcoholic media, liposome suspensions, powders, lotions, milks, creams, salves, gels, foams and ointments.

22. The method according to any one of claims 14 to 21, wherein the cosmetic composition is selected from a cosmetic product, a makeup product or a body hygiene product, for example a lotion, a milk, a serum, an aqueous or oily gel, an emulsion, a cream, in particular a day cream or a night cream, a cream-gel, a skincare water, an ointment, a balm, a foundation, a spray, an eyeshadow, a stick, a lipstick, a gloss, a lip balm, a foam, a deodorant, a nourishing mask, a shower gel, and an exfoliating or scrubbing product.

23. A composition comprising: - from 20% to 40% by weight of a liquid extract obtained from an aerial part of Anigozanthosflavidus as an anti-agening agent; and - from 60% to 80% by weight of glycerol.

24. Use of the composition of claim 23 in the manufacture of a cosmetic composition for preventing or treating skin ageing.

25. A method of treating skin ageing in a subject including the step of applying to the skin of said subject an effective amount of a composition according to claim 23.