HK1204930B - Cosmetic use of an albizia julibrissin extract and corresponding topical composition - Google Patents

Description

Cosmetic use of albizzia julibrissin extracts and corresponding topical compositions

Technical Field

The present invention discusses the cosmetic topical application of the extract of Albizzia Julibrissin and corresponding topical compositions.

The present invention relates generally to the cosmetic and dermopharmaceutical industries; these industries produce and/or use products for treating mammalian, animal or human skin (including scalp), mucous membranes and appendages (such as body hair, eyelashes, eyebrows, nails or hair) with the aim of improving their appearance and/or general condition.

The demand of these industries for new products is increasing, in particular the need for active ingredients extracted from plants; as they allow for a combination of efficiency, limit risk of irritation and allergy, reduce side effects, biodegradability, have a chance to mark/prove, and applicability to sustainable development and/or fair trade methods.

The present invention aims to meet this need.

Albizzia julibrissin is also known as "Silk Tree" or "Mimosa of Constantinople", a deciduous Tree belonging to the family Mimosaceae. Its origin is in east asia and south america, but has already been spread by humans to almost every continent.

Albizzia julibrissin extract has been proposed for use in cosmetics.

JP2009242296 discloses albizia julibrissin extracts capable of inhibiting the effect on melanin production and stimulating collagen production.

KR20100090530 discloses the use of an extract of albizia julibrissin dura to inhibit the activity of metalloproteinases.

KR20020080657 discloses the anti-radical activity of albizia julibrissin dura extract.

JP4342519 discloses the use of albizzia julibrissin extract to inhibit tyrosinase activity.

JP2000143488 discloses the wettability of albizzia julibrissin extract on skin and hair.

JP2048515 discloses the use of albizzia julibrissin extracts in hair products for preventing hair loss and promoting growth.

JP2010235548 discloses an agent used in cosmetics and foods/beverages for whitening skin, restoring hair, providing antioxidant effect and treating diseases (e.g. aging, obesity and inflammation); the agent comprises an extract of Albizzia julibrissin.

Disclosure of Invention

The subject of the present invention is the use of an extract of albizzia julibrissin for the topical cosmetic treatment of glycation.

The human body needs sugar, glucose, to produce energy for it. However, glucose can never be fully used by the human body for this advantageous purpose. The remainder of the absorbed sugar will react with the amine groups of the protein, nucleic acid or lipid in a non-enzymatic manner to form a highly glycosylated end product (referred to as AGE product for short).

AGE formation involves several successive stages. In the early stages, the aldehyde group of reducing sugars (such as glucose) forms an unstable bond with protein amines, known as the "schiff base"; it rearranges rapidly to produce amadori compounds that are not in the final state. This amadori product undergoes many subsequent reactions such as oxidation, condensation, dehydration, formation of intermediate reagents (e.g., aldehydes), leading to the aggregation of complex end products of Advanced Glycation (AGEs) that aggregate on tissues over time.

In the amadori product stage, sugars can also be deprotonated to form several very reactive molecules: glyoxal (GO) or its methylglyoxal derivative (MGO), 3-deoxyglucosone (3-DG) or fructosamine. Glyoxal and 3-DG can then bind to free protein amines to form different AGEs, such as carboxymethyl-lysine (CML), carboxymethyl-arginine (CMA), carboxymethyl-cysteine (CMC) and their ethyl equivalents or pyrrolin, methylglyoxal-lysine dimer, and the like.

The formation of these same reactive chemical forms can also be performed as a result of glucose autooxidation, resulting in dicarbonylated glycotoxins that can readily react with glyoxal, methylglyoxal, and 3 deoxyglucosone types; these molecules mentioned above can lead to the synthesis of AGEs.

In addition, other sugars (such as ribose or xylose), although present in smaller quantities, are much more reactive than glucose and fructose and also form AGEs; the in vitro toxicity of AGEs has actually been demonstrated.

These different side reactions lead to the formation of many improved proteins (glycation); their functional, enzymatic and structural properties are impaired, with serious consequences for the good functioning of a cell or organism. In addition, binding of nucleic acids to glycotoxins results in mutations that may be harmful to the cell.

In the case of the skin, AGEs are found not only in the dermis, but also in the epidermis, as far as the stratum corneum. The effects on organs are more severe if their proteins have a long life span. Thus, dermal collagen with a half-life of 15 years is 3 times more abundant in carboxymethyl-lysine (CML), N-carboxyethyl-lysine (CEL), and pentose sugars at 20 years of age compared to 80 years of age. The properties of collagen, as well as those of elastin, fibronectin and laminin, are altered, leading to changes in the mechanical and elastic properties of the dermal extracellular matrix; the dermis becomes less flexible and more rigid and also looser and less active. Indeed glycation does not only affect proteins with a long lifetime. Vimentin is an intracellular protein with a much shorter lifespan than collagen. It is an important synthetic part of the intermediate filaments of the cytoskeleton. Vimentin is involved in many functions of skin fibroblasts, such as healing, fluidity, and contraction. AGEs formed from MGO accumulated in vimentin cause it to aggregate, reduce the contractile capacity of cells, and disorganize collagen fibers.

Inside the cell, mitochondrial proteins are directly and indirectly attacked by glucose, which is not used for energy production. These proteins are glycated, resulting in a decrease in ATP synthesis yield and defective energy production. This process occurs in all body cavities, including the skin. Cells also have metabolic sensors or probes; they allow to regulate their energy production. AMPK (AMP-activated protein kinase) is one of them. This responds to reduced ATP by increasing ATP biosynthesis. The active site of the enzyme is known to be very sensitive to saccharification; thus, it may lose its regulatory function.

Furthermore, AGEs alter vascular permeability through glycation/alteration of their matrix proteins. The ensuing micro-inflammatory reaction causes the blood vessel flow to decrease and blood components to leak out of the blood vessel, causing the accumulation of fluids and waste products around them. This results in the formation of dark circles and some forms of bags under the eyes.

AGEs are therefore a significant source of cellular dysfunction; moreover, their gradual accumulation tends to reduce the properties and performance of tissues and cells: less flexibility, less mobility, less responsiveness, less energy production. All functions are impaired.

In addition, the cells have an active defense mechanism against glycation based on an enzyme system, called glyoxalase-1 and-2; through intervention of GSH, it detoxifies methylglyoxal and other reactive aldehyde species. Saccharification products tend to reduce glyoxalase activity. As mentioned above, saccharification is a rapid effect; its involvement in various organs can be quantified after only a few days of hyperglycemia. Glycotoxins result in a decrease in the ability of cells to produce energy and macromolecular changes. The effects of glycotoxins may also accumulate over time. They result in a decrease in the responsiveness of cells to additional stress and result in a "cell fatigue" that can be understood by simulation using physical fatigue. When the cell (or body) is young, its recovery capacity is maximal and it recovers fastest. The accumulation of daily fatigue reduces this ability and amplifies the deleterious effects of glycotoxins.

Transient fatigue becomes commonplace and then becomes chronic. These signs can be seen on the face: dark under-eye circles and bags, dull and elongated facial features, less lively skin, tired skin. In cells and tissues, metabolism is impaired.

Therefore, fighting cell fatigue by reducing the daily impact of glycotoxins is one of our goals to be achieved, in particular by protecting melatonin; melatonin is a molecule produced in different parts of the body, especially in the skin. It has an effect on the compensatory regulation and protection of sleep, especially by protecting the energy production capacity of the cells. The glycotoxin can reduce melatonin level; while maintaining these levels has a direct benefit in protecting the restoring capacity of the cells.

The effects of these biochemical processes on the skin are so important; we have seen their effect at the cellular level. Glycation results in dull skin, loose skin, lack of radiance, a tired appearance, e.g. dark under eye circles and bags, lack of tension and flexibility.

The applicant has shown that albizia julibrissin extract according to the present invention exhibits two decisive and complementary effects against glycation:

first, to combat the preventive effect of glycation, to prevent its implementation;

second, a repairing effect, referred to according to the invention as "desulphatation" effect or "desulphation", or detoxification effect; in case saccharification has already started, it allows to limit its negative consequences. The saccharified product is neutralized or detoxified by the albizia julibrissin extract, resulting in a healing effect on tired skin, especially with loss of gloss and softness.

Thus, in accordance with the present invention, albizzia flower extract can be used, inter alia:

treatment of skin fatigue (exhausted skin); and/or

-detoxification of the skin (including the scalp) and its appendages; and/or

-improving the brightness or luminosity of the complexion of the skin and its appendages and treating the loss of skin suppleness; and/or

For treating eye contours, in particular dark circles and bags.

In vivo and in vitro assays, described in detail below, demonstrate the glycation of skin molecules, particularly proteins, by Albizzia julibrissin extractsThe activity of the substance. In particular, AGE-Reader^TM(DiagnOptics Technologies Inc.) is used as a medical device for the non-invasive measurement of AGEs accumulation.

According to the invention, an "extract" is an extract which can be obtained by means of conventional extraction techniques or by means of in vitro plant cell culture. For example, common techniques that can be used include soaking, simple decoction, leaching, reflux extraction, supercritical fluid extraction, ultrasonic or microwave extraction, or extraction using counter-current techniques. The extraction solvent can be selected from the following: water, propylene glycol, butylene glycol, glycerol, PEG-6 caprylic/capric glycerides, polyethylene glycol, methyl and/or ethyl diglycol ethers, cyclic polyols, ethoxylated or propoxylated diglycols, alcohols (methanol, ethanol, propanol, butanol); or any mixture of these solvents.

The invention also covers an extract obtained by in vitro plant culture. Various techniques exist, including culture of undifferentiated or dedifferentiated cells, tissue or organ culture, or in vitro micropropagation by somatic embryogenesis or by vegetative propagation.

The extracts obtained by in vitro plant culture exhibit advantages compared to the agro-industrial route (plant culture in open field, subsequent extraction in the factory): the obtained material is free from toxic substances (herbicides, etc.), reproducibility is improved, biodiversity is retained, and the like.

According to the invention, all parts of the plant, including aerial (such as leaves, flowers or seeds or roots), can be used.

According to the present invention we prefer albizzia extract obtained from the flowers and/or seeds of the plant; the specific description is as follows.

According to the present invention, the albizzia julibrissin extract can be used neat or diluted in a physiologically acceptable excipient or matrix.

The extract is preferably applied by dilution in a physiologically acceptable excipient or matrix, especially a hydroalcoholic matrix.

The present invention also encompasses topical compositions, i.e., an effective amount of albizzia julibrissin extract in a physiologically acceptable medium, included as an active ingredient for saccharification. As explained above, the albizzia julibrissin extract has a significant effect on the desugarization of proteins and skin detoxification.

The topical application may be a cosmetic and/or a dermopharmaceutical.

According to other advantageous features, the albizzia julibrissin extract can be used in combination with one or more additional active ingredients, in particular to provide a broader range of cosmetic properties. For example, the additional active ingredients may be selected within the following ranges: whitening, anti-redness, anti-spotting active ingredients, calming active ingredients, active ingredients for treating allergy, reactive skin, UV sunscreen, moisturizing, moisturizers, exfoliating, smoothing, toning, anti-aging, anti-wrinkle and fine lines, active ingredients for improving mechanical properties and elasticity, complexion luminosity, detoxification, anti-hair regrowth active ingredients, active ingredients acting on the skin barrier, anti-acne active ingredients, active ingredients acting on sebum secretion, matting, homogenization, anti-inflammatory, anti-oxidant, anti-radical, anti-glycation active ingredients, active ingredients for eye contour (dark circles and under-eye bags), active ingredients for enhancing activity, active ingredients for promoting blood circulation, peptides, vitamins, etc. These active ingredients can be obtained from plant materials, such as plant extracts, or products obtained by plant culture or fermentation.

Surprisingly, the inventors have found that the albizzia julibrissin extract of the invention, when combined with siegesbeckia orientalis glycosides, is particularly suitable for treating the face and more particularly the eye contour. According to the invention regarding the overall eye contour treatment (preventive and therapeutic), siegesbeckia orientalis glycosides potentiate and complement the anti-glycation activity of albizzia julibrissin.

The siegesbeckia glycosides are in the form of a sugar of siegesbeckia alcohol and have the following formula:

siegesbeckia glycosides can be obtained from any source; in particular by means of semi-chemical synthesis, enzymes, by one of the many methods of biotechnology, by plant extraction, or by any other means used to obtain it at a reasonable cost in the end product used industrially.

According to the specific equipment, the extract is vegetable Siegesbeckia Orientalis glycoside, which is extracted from Siegesbeckia Orientalis; this plant is also known as "holy herb" or "st. paul's worth". Although native to India, Siegesbeckiae herba has been spread to many tropical countries, especially to Monga. It is known as a medicinal plant with calming and cicatrisation effects. FR2285142 discloses a method for obtaining siegesbeckia glycosides from Siegesbeckiae herba.

The siegesbeckia glycosides can also be present in the form of an extract of siegesbeckia orientalis, including titrated amounts of siegesbeckia glycosides.

Siegesbeckiae herba glycosides are added into different cosmetic materials. For example:

siegesbeckiae herba glycosides sold by Sederma corporation^TM: molecular Siegesbeckiae herba extract of Siegesbeckiae herba and extract combination of Centella Asiatica (Centella Asiatica) rich in asiaticoside; centella asiatica has anti-inflammatory and remodelling properties on the dermal extracellular matrix, and this component has been claimed to be useful in the treatment of striae during pregnancy.

Chromocare sold by Sederma corporation^TM: a combination of siegesbeckia orientalis extract enriched in siegesbeckia orientalis glycoside and Rabdosia rubescens extract enriched in rubescensine; it can combat oxidative stress and beneficially affect chromophore aging (collagen degradation, inflammation and melanin stimulation).

According to the present invention, a composition comprising albizzia julibrissin and siegesbeckia orientalis extract in a physiologically acceptable medium is provided.

In summary, the albizzia julibrissin extract is an extract obtained according to the present invention.

In vitro tests detailed in the specifications show the activity and the best synergy obtained with the composition of albizzia julibrissin extract and siegesbeckia orientalis glycoside according to the invention. In particular, it has been shown that:

-an increase in an anti-aging gene;

-a synergistic effect on the non-enzymatic glycation of elastin;

-inhibitory activity of metalloproteinases MMP1 post-stress;

-an increase in collagen I and elastin.

In vivo tests, also detailed below, show the cosmetic advantages of the albizzia julibrissin extract and siegesbeckia orientalis glycoside composition in terms of visible global fatigue of the eye contour: fine lines and wrinkles, eyelids, dark circles and bags.

The other additional active ingredients can be chosen in particular from the following ranges: vitamin B3 compounds, nicotinamide or tocopheryl compounds, retinoid compounds, such as rosin oil, hexamidine, alpha-lipoic acid, resveratrol or DHEA, peptides, in particular N-acetyl-tyrosine-arginine-O-hexadecyl ester, Pal-VGVAPG (SEQ ID NO: 1), Pal-KTTKS (SEQ ID NO: 2), Pal-GHK, Pal-KMO2K and Pal-GQPR (SEQ ID NO: 3); they are conventional active ingredients used in topical cosmetic or dermal pharmaceutical compositions.

The present invention will be better understood from the following description.

DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION

Preparation of the composition

According to the invention, the expression "physiologically acceptable medium" means, without limitation, an aqueous or hydroalcoholic solution, a water-in-oil emulsion, an oil-in-water emulsion, a micro-emulsion, an aqueous gel, an anhydrous gel, a slurry, vesicles or a dispersion of a powder.

By "physiologically acceptable" is meant that the composition is suitable for epidermal or transdermal use, and may contact the mucosa, appendages (nails, hair, body hair), scalp and skin of mammals, especially humans, without further risk of toxicity, incompatibility, instability, allergic response, and the like.

This "physiologically acceptable medium" constitutes what is commonly referred to as a synthetic excipient.

For use according to the present invention, the effective amount of albizzia julibrissin extract or siegesbeckia orientalis glycoside, if present, that is, its dosage, depends on various factors such as age, the skin condition of the patient, the severity of the condition or illness, and the mode of administration, among others. By effective amount is meant a non-toxic amount sufficient to achieve the desired effect.

For anti-glycation and eye contour applications according to the present invention, the albizzia julibrissin extract or siegesbeckia orientalis glycoside, if present, is generally present in an effective amount ranging from 0.00001% to 90%, more particularly from 0.0001% to 25%, and most particularly from 0.001% to 10%; based on the total weight of the composition. The person skilled in the art is able to adjust the amount of extract according to the desired effect. All percentages and ratios used herein refer to the weight of the total composition and all measurements are made at 25 ℃; unless otherwise specified.

The excipients for the composition are selected according to the constraints of the materials of albizzia and siegesbeckia orientalis glycosides, if present (stability, solubility, etc.), and, if necessary, according to the dosage form that is further intended for use in the composition.

When combined into a composition, the albizzia julibrissin extract and siegesbeckia orientalis glycoside (if present) can be dissolved in the composition with the aid of an aqueous solution, or by means of a common physiologically acceptable co-solvent, such as (but not limited to) this list: ethanol, propanol, isopropanol, propylene glycol, glycerol, butylene glycol, or polyethylene glycol, or any combination thereof. It may also be of interest to use emulsifiers to solution extract. Powder media can also be used.

The compositions useful in the present invention are generally prepared by conventional methods well known to those skilled in the art of making topical and oral compositions and injection compositions. Such methods may involve compounding a mixture in one or more steps to obtain a homogeneous state, with or without heating, cooling, etc.

The different galenical forms which may contain the albizzia julibrissin extract and the siegesbeckia orientalis glycosides, if present, may be all those listed below: for example, creams, lotions, emulsions or creams, gels, lotions, dispersions, solutions, suspensions, cleansers, foundations, anhydrous preparations (in stick form, in particular lip balms, body and bath oils), shower and bath gels, shampoos and hair lotions, creams or lotions for skin care or hair, cleansing emulsions or emulsions, sun-screening emulsions, lotions or emulsions, artificial tanning emulsions, emulsions or emulsions, pre-, mid-or after-shave emulsions, foams, gels or lotions, cosmetics, lipsticks, mascara or nail varnish, skin balms, serums, adhesives or adsorbing materials, transdermal patches, or powders, lotions, emulsions or emulsions, sprays, body and bath oils, foundations, ointments, emulsions, colloids, pressed suspensions or solids, pencils, applied formulations, brushable, blush, red, eye-depicting creams, lip pencils, lip balms, facial or body powders, styling gels or mousses, nail conditioners, lipsticks, skin conditioners, moisturizers, glossers, soaps, exfoliants, astringents, depilatories, permanent wave solutions, antidandruff formulations, antiperspirant or antiperspirant compositions, including fine sticks, "roll-on" deodorants, air fresheners, nasal sprays, and the like. These compositions may also take the form of lipsticks, the purpose of which may be to colour the lips or to prevent them from chapping, or eye make-up, eye shadow and face primer. The compositions of this patent may include cosmetic, personal care products and pharmaceutical formulations. It is also contemplated to include a pressurized propellant, a composition in the form of a foam, or a composition in the form of an aerosol. The composition according to the invention can also be used for oral applications, such as toothpaste. In this case, the composition may contain adjuvants and additives commonly used in oral applications, including surfactants, thickeners, humectants, polishing agents (such as silica), various active ingredients such as fluoride, especially sodium fluoride, and optionally sweeteners such as sodium saccharin.

The form of the albizzia julibrissin extract and siegesbeckia orientalis glycoside, if present, can be a solution, suspension, emulsion, paste, or powder, either alone or as a premix, or in a separate carrier or vehicle, such as a macrocapsule, a micro-or nanocapsule, a macrosphere, a micro-or nanosphere, a liposome, an oleosome or chylomicron, a macro-, micro-, or nanoparticle, or a macrosponge, a micro-or nanoemulsion, or an adsorbed organic polymer powder, talc, bentonite, spores or adventitia, as well as other inorganic or organic carriers, according to the present invention.

The albizzia julibrissin extract and siegesbeckia orientalis glycoside (if present) can be used in any form according to the present invention; in the form of a macro-, micro-, and nano-particle, or macro-, micro-, and nano-coating, bound to or incorporated into, or adsorbed onto, a textile, natural or synthetic fiber, wool; and any material that can be used in daily or night use cloths or underwear, handkerchiefs or clothes intended to be in direct contact with the skin, so as to exert their cosmetic effect by such skin fabric contact and allow continuous local release.

Cosmetic treatment method

The present invention also provides a method of cosmetic topical treatment for treating glycation of skin proteins, comprising topically applying to the skin of a subject in need of such treatment an effective amount of a composition comprising an extract of albizia julibrissin as described above.

By "topical treatment" is meant a coating intended to be used where it is applied: skin, mucosa, skin appendages.

The method according to the invention is particularly suitable for:

-treating exhausted skin (skin fatigue) which has lost their luminosity and softness; and/or

-detoxification of the skin (including the scalp) and its appendages; and/or

-improving the brightness of the complexion of the skin and its appendages and managing the loss of skin softness; and/or

For processing eye contours, in particular black eye circles and bags.

According to other features of the method, the albizzia julibrissin extract can be conveniently combined with siegesbeckia orientalis glycosides, as described above; this method is therefore particularly suitable for treating exhausted skin, especially for eye contours caused by the synergistic effect of dark circles and bags, wrinkles and fine lines, and eyelid movement.

By periodic topical application, such as daily, improvements in the appearance and general condition of the skin, mucous membranes and appendages can be obtained.

A practitioner treats a topical cosmetic comprising a composition comprising an extract of albizzia julibrissin; such treatment may be achieved, for example, by topically applying the compositions described in this patent, according to the methods commonly used for applying such compositions. The topical composition is preferably applied once daily for a period of at least one week; however, it can also be applied over a period of 2, 4, 8 or 12 weeks. The topical composition is preferably applied to the face and neck, but can be applied to any area of the skin where aesthetic improvements are desired; here, the composition remains on the area of skin that needs to be treated and preferably is not removed or washed away from the skin. For reference, the European standard dosage of skin cream for cosmetic facial treatment is 2.72mg/cm²Day/person.

It is also to be understood that as used herein, the terms treatment and treatment include and encompass the reduction, amelioration, advancement, release, and/or elimination of a dermatological effect, including aging. The compositions and methods of the present invention are suitable for treating skin conditions in many areas on the skin, including (but not limited to) the face, forehead, lips, neck, neckline, arms, hands, body, legs, knees, feet, chest, back, buttocks, and the like.

One of the important advantages of the present invention is that by this local, non-invasive method of application, a locally selective "gentle" treatment can be applied at any time necessary or desired. For anti-wrinkle applications, for example, a very topical application may be performed using a syringe or micro-sleeve.

However, it is also conceivable to inject the composition according to the invention subcutaneously.

According to other features, the cosmetic treatment method of the invention can be combined with one or more other treatments targeted to the skin, such as photo-therapy, thermal or aromatherapy treatments.

According to the invention, a device with several compartments or kits can be proposed to apply the above method; for example, they may include, but are not limited to, a composition comprising an extract of Albizzia julibrissin of the present invention in a first compartment and a composition comprising other active ingredients, such as siegesbeckia orientalis glycosides, and/or excipients in a second compartment. In this case, the compositions contained in the first and second compartments are considered to be combined compositions for simultaneous, separate or stepwise use in due course, in particular in one of the above-cited treatment methods.

Additional ingredients

CTFA international Cosmetic ingredient dictionary and handbook (revised 13, 2010) (publishers are Cosmetic, Toiletry, and Fragrance Association, inc. located in washington, d.c. usa) describes unlimited, very wide range of Cosmetic and pharmaceutical ingredients that are routinely used in the skin care industry; they can be used as additional ingredients/compounds in the compositions of the present invention. Examples of these ingredient classes include, but are not limited to: healants, anti-aging agents for the skin, anti-wrinkle agents, anti-atrophy agents, skin moisturizers, skin smoothing agents, antibacterial agents, antiparasitic agents, antifungal agents, bacteriostatic agents, antimicrobial agents, anti-inflammatory agents, anti-itch agents, anesthetic agents, antiviral agents, keratolytic agents, radical scavengers, seborrhea agents, antidandruff agents, skin differentiation, proliferation or pigmentation modulators, permeation accelerators, desquamatories, melanin synthesis stimulators or inhibitors, whitening, bleaching or lightening agents, protein colorants, self-tanning agents, NO-synthase inhibitors, antioxidants, radical scavengers and/or anti-atmospheric agents, active carbonyl species scavengers, anti-glycation agents, tightening agents, agents that stimulate the synthesis of epidermal or epidermal macromolecules, and/or agents that inhibit or prevent their degradation, such as collagen synthesis stimulators, an elastin synthesis stimulator, a decorin synthesis stimulator, a laminin synthesis stimulator, a defensin synthesis stimulator, a chaperonin synthesis stimulator, an aquaporin synthesis stimulator, a hyaluronic acid synthesis stimulator, an fibronectin synthesis stimulator, a longevity gene protein synthesis stimulator, a synthesis stimulator of lipids and stratum corneum components (ceramides, fatty acids, etc.), a collagen degradation inhibitor, an elastin degradation inhibitor, a fibroblast proliferation stimulator, a keratinocyte proliferation stimulator, a adipocyte proliferation stimulator, a melanocyte proliferation stimulator, a keratinocyte differentiation stimulator, an adipocyte differentiation stimulator, an acetylcholinesterase inhibitor, a glycosaminoglycan synthesis stimulator, a DNA repair agent, a DNA protectant, an antipruritic, a sensitive skin treatment and/or care agent, a solidifying agent, anti-striae gravidarum, astringents, sebum secretion regulators, dermal relaxants, adjunctive therapeutic agents, re-epithelialization stimulants, re-epithelialization coadjuvants, cytokine growth factors, sedatives, anti-inflammatory agents, agents acting on capillary circulation and/or microcirculation, angiogenesis stimulants, vascular permeability inhibitors, agents for cell metabolism, agents for improving dermal-epidermal junction, agents for inducing hair growth, agents for inhibiting or retarding hair growth, muscle relaxants, anti-pollution and/or anti-radical agents, agents for stimulating lipolysis, agents for reducing body weight, anti-cellulite agents, agents for promoting microcirculation, agents for cell energy metabolism, cleansing agents, hair regulators, hair styling agents, hair growth promoters, sunscreens, complete sunscreens, make-up agents, detergents, drugs, emulsifiers, emollients, organic solvents, preservatives, deodorant active ingredients, physiologically acceptable carriers, surfactants, abrasives, adsorbents, aesthetic components such as perfumes, pigments, dyes, colorants, natural colorants, essential oils, touch agents, cosmetic astringents, anti-acne agents, anticoagulants, antifoams, antioxidants, binders, biological additives, enzymes, enzyme inhibitors, enzyme inducers, coenzymes, chelating agents, plant extracts, plant derivatives, essential oils, marine extracts, agents obtained from biofermentations or biotechnological processes, mineral salts, cell extracts, sunscreens (organic or mineral photoprotective agents effective against uv a and/or B rays), ceramides, peptides, buffers, volume enhancers, chelating agents, chemical additives, colorants, cosmetic biocides, denaturants, medical astringents, external analgesics, film formers, such as polymers, for enhancing the film forming and substantivity of the composition, quaternary ammonium derivatives, substantivity enhancing agents, sunscreens, pH adjusting and regulating agents (e.g., triethanolamine), propellants, reducing agents, sequestering agents, depigmenting and/or lightening agents, skin conditioning agents (e.g., moisturizers, including miscellaneous and occlusive), moisture retention agents, alpha-oxo acids, beta-oxo acids, moisturizers, epidermal hydrolases, healing and/or soothing agents, skin treatment agents, anti-wrinkle agents, pouch reduction or treatment agents, exfoliants, thickening agents, emollients, gelling polymers, vitamins and their derivatives, humectants, exfoliants, tranquilizers, skin treatment agents, lignans, preservatives (e.g., phenoxyethanol and parabens), anti-uv agents, cytotoxic agents, anti-neoplastic agents, viscosity modifiers, non-volatile solvents, pearlescent agents, antiperspirants, depilatories, vaccinia, perfume, skin restructuring agents (e.g., siegesbeckia orientalis extract), excipients, fillers, minerals, antimycobacterial agents, antiallergic agents, H1 or H2 antihistamines, anti-irritants, immune system stimulants, immune system suppressants, insect repellents, lubricants, pigments or dyes, discolorants, preservatives, light stabilizers, and mixtures thereof; provided that they are physically and chemically compatible with the other ingredients of the composition, in particular with the active ingredients of the invention. Also, the nature of these additional ingredients must not cause unacceptable changes to the active ingredients of the present invention. These additional ingredients may be synthetic or natural, such as plant extracts, or from biological fermentation processes. Further examples can be found in the CTFA handbook of cosmetic ingredients.

Such additional active ingredients/compounds may be selected from the whole group including: sugar amines, glucosamine, D-glucosamine, N-acetylglucosamine, N-acetyl-D-glucosamine, mannosamine, N-acetylmannosamine, galactosamine, N-acetylgalactosamine, vitamin B3 and derivatives thereof, niacinamide, sodium dehydroacetate, dehydroacetic acid and salts thereof, phytosterols, salicylic acid compounds, hexamidine, dialkanoyl hydroxyproline compounds, soybean extracts and derivatives, equol, isoflavone, flavone, phytantriol, farnesol, geraniol, bisabolol, peptides and derivatives thereof, di-, tri-, tetra-, penta-and hexapeptides and derivatives thereof, KTTKS (SEQ ID No. 4), Pal-KTTKS (SEQ ID No. 2), carnosine, N-acylamino acid compounds, retinoids, retinylpropionic acid, rosin oil, retinol palmitate, retinyl acetate, retinal, retinoic acid, water-soluble vitamins, ascorbic acid, vitamin C, ascorbyl glucoside, ascorbyl palmitate, magnesium ascorbyl phosphate, sodium ascorbyl phosphate, vitamin B and salts and derivatives thereof, vitamin B1, vitamin B2, vitamin B6, vitamin B12, provitamin and salts and derivatives thereof, vitamin K and derivatives thereof, pantothenic acid and derivatives thereof, panthenyl ethyl ether, panthenol and derivatives thereof, panthenol, biotin, amino acids and salts and derivatives thereof, water-soluble amino acids, asparagine, alanine, indole, glutamic acid, water-insoluble vitamins, vitamin A, vitamin E, vitamin D and mono-, di-and tri-terpenoids, beta-ionol, cedrol and derivatives thereof, water-insoluble amino acids, tyrosine, tryptamine, particulate materials, butylhydroxytoluene, butylhydroxyanisole, allantoin, tocopherol nicotinate, tocopherol esters, palmitoyl-Gly-His-Lys (Pal-GHK), phytosterols, hydroxy acids, lactic acid, lactobionic acid, keto acids, pyruvic acid, phytic acid, lysophosphatidic acid, stilbene, cinnamate, resveratrol, kinetin, zeatin, dimethylaminoethanol, natural peptides, soy peptides, salts of sugar acids, manganese gluconate, zinc gluconate, piroctone amine, 3,4,4' -trichlorocarbanilide, triclocarban, zinc pyrithione, hydroquinone, kojic acid, ascorbic acid, magnesium ascorbyl phosphate, ascorbyl glucoside, pyridoxine, aloe vera, terpene alcohol, allantoin, bisabolol, dipotassium glycyrrhizinate, glyceric acid, sorbitic acid, pentaerythritolic acid, pyrrolidone carboxylic acid and its salts, dihydroxyacetone, erythrulose, glyceraldehyde, clove oil, menthol, camphor, eucalyptus oil, eugenol, menthyl lactate, witch hazel distillate, eicosene and vinylpyrrolidone copolymers, iodopropyl carbamate, polysaccharides, essential fatty acids, salicylates, glycyrrhetinic acid, carotenoids, ceramides and pseudoceramides, lipid complexes, oils generally of natural origin, such as shea butter, kernel oil, evening primrose oil, plum oil, palm oil, gardenia oil, hydroquinone, HEPES, propylcysteine, O-octanoyl-6-D-maltose, the disodium salt of methylglycine diacetic acid, steroids, such as diosgenin and derivatives of DHEA, DHEA or dehydroepinone and/or their precursors or chemical or biological derivatives, n-ethoxycarbonyl-4-p-aminophenol, extracts of blueberries, phytohormones, extracts of saccharomyces cerevisiae, extracts of seaweed, extracts of soya beans, lupins, maize and/or peas, alverine and its salts, in particular alverine citric acid, extracts of butcher's broom and horse chestnut, octadecadienoic acid, their analogues and derivatives, and mixtures thereof, metalloproteinase inhibitors.

More skin care and hair care actives that are particularly useful in combination with the composition can be found in the "SEDERMA Business literature" and the Web sitewww.sederma.frFind in.

For example, the following commercially active ingredients may also be mentioned: betaine, Glycerol, Actimoist Bio 2^TM(Activeorganics) (comments: trade name and company name), AquaCaptec^TM(Mibelle AG Cosmetics)，Aquaphyline^TM(Silab)，AquaregulK^TM(Solabia)，Carciline^TM(Greentech)，Codiavelane^TM(Biotech Marine)，Dermaflux^TM(Arch Chemicals，Inc)，Hydra'Flow^TM(Sochibo)，Hydromoist L^TM(Symrise)，RenovHyal^TM(Soliance)，Seamoss^TM(Biotech Marine)，Essenskin^TM(Sederma)，Moist 24^TM(Sederma)，Argireline^TM(trade name of acetyl hexapeptide-3, Lipotec Co., Ltd.) known under the name Gatuline Expression^TMExtracts of spilanthol or Acmella oleracea known under the name Boswellin^TMThe extract of the lower mastic tree, Deepaline PVB^TM(Seppic)，Syn-AKE^TM(Pentapharm)，Ameliox^TM，Bioxilift^TM(Silab)，Juvinity^TM(Sederma)，Revidrate^TM(Sederma)，Chronodyn^TM(Sederma)，O.D.A.white^TM(Sederma)，Idealift^TM(Sederma)，Resistem^TM(Sederma)，Widelash^TM(Sederma) or mixtures thereof.

Among other plant extracts that can be combined with the composition of the invention, there may be extracts that specifically mention the following: ivy, in particular ivy (Hedera Helix; trade mark: an alternative name with the same meaning in brackets for Chinese), Bupleurum root, Stellaria dichotoma, Arnica Montana L, Rosmarinus officinalis N, marigold Calendipa L, Salvia officinalis L, Panax ginseng P.G., Ginkgo biloba L, Hypericum Perforatum L, Ruscus aculeatus L, spiraea europaea Lndiula ulmaria L), java tea (orthophon standardization Benth), seaweed (fucus vesiculosus), birch (Betula alba), green tea, kola nut (Cola), horse chestnut, tobacco, centella asiatica, photinia, fucus, willow, gnaphalium, aescine, atractylodes, chamomile, apricot, atractylodes, platycodon grandiflorum, simenum, petunia, caraway, plant of the genus euphorbia, humifusa purpurea, such as c.forskohlii, c.blumei, c.esquiroliii, c.scutulareoides, c.xanthanthus and c.bautus, such as the root extract of carnation; ballote, Guioa, Davallia, Terminalia, Barringtonia, Trema, antiarobia, bugle, argania, extracts of yam, such as dioscorea opposita or dioscorea mexicana; ammi Pumila, extract of Siegesbeckiae herba, especially Siegesbeckiae herba, vegetable extract of Ericaceae, especially cowberry fruit extract (Vaccinium angustifolium) or Arctostaphylos uva ursi, aloe vera, sterol-containing plant (such as phytosterol), Manjisha (extracted from Rubia plants, especially Rubia cordifolia), and Mucurel (extracted from Commiphora plants, especially Commiphora myrrha), Cola fruit extract, flos Matricariae Chamomillae, Trifolium Pratentis L.extract, Piper methysticum extract (Kava of SEDERMA corporation)^TM) Bacopa monnieri extract (Bacocalmine from SEDERMA)^TM) And sea penis extract; licorice, mulberry, cajeput (tea tree), acanthus mollis, rabdosia rubescens, euglena, hirudo of aristolochia, charantia sorghum, sunflower extract, eucalypt, mitracepe of the genus stermococea, buchurosma, henna, adiium capitatum-Veneris l., greater celandine, luffa, japanese citrus (citrus variety of satsuma mandarin), chongqing loquat tea, imperata, poppy of the yellow sea, cedar, polygonum odoratum, hemsleya macrosperma, sambucus nigra, gossypol, all manis, kelp, terna, rhizoma anemarrhenae, purslane, hops, arabica coffee tree, yerba mate, zingiber zerumbet Smith, globus cordifolia, or ulva extract.

The compositions of the present invention may include peptides, including but not limited to di-, tri-, tetra-, penta-, and hexapeptides, and derivatives thereof. The concentration of the additional peptide in the composition ranges from1x10^-7% to 20%, preferably from 1X10^-6% to 10%, preferably 1x10^-5% and 5%; all are in weight ratio.

According to the present invention, the term "peptide" refers to various peptides containing 10 amino acids or less, their derivatives, isomers, and complexes with other species, such as metal ions (e.g., copper, zinc, manganese, magnesium, etc.). The term "peptide" refers to both natural and synthetic peptides. It also refers to compositions containing naturally-found, and/or commercially-available peptides.

Suitable dipeptides for use herein include, but are not limited to, carnosine (β -AH), YR, VW, NF, DF, KT, KC, CK, KP, KK, or TT. suitable tripeptides for use herein include, but are not limited to, RKR, HGG, GHK, GKH, GGH, GHG, KFK, KPK, KMOK, KMO₂K or KAvaK. Suitable tetrapeptides for use herein include, but are not limited to, RSRK (SEQ ID NO: 5), GQPR (SEQ ID NO: 6) or KTFK (SEQ ID NO: 7). Suitable pentapeptides for use herein include, but are not limited to, KTTKS (SEQ ID No.: 4). Suitable hexapeptides include, but are not limited to, GKTTKS (SEQ ID NO: 8), VGVAPG (SEQ ID NO: 9).

Other suitable peptides for use herein include, but are not limited to, lipophilic derivatives of the peptide, most preferably palmitoyl derivatives, and the metal complexes mentioned above (e.g., copper complex of the tripeptide HGG.) most preferred dipeptide derivatives include N-palmitoyl- β -alanine-Ala-His, N-acetyl-tyrosine-arginine-hexadecyl ester (Calmosense)^TM，Idealift^TMFrom Sederma corporation). Preferred tripeptide derivatives include N-palmitoyl-Gly-Lys-His, and Pal-Gly-His-Ly, (Pal-GKH and Pal-GHK from Sederma), the copper derivative of HGG (Lamin)^TMFrom Sigma), Lipospondin (N-Elaidoyl-KFK) and its conservatively substituted analogues, N-acetyl-RKR-NH 2 (peptide CK +), N-Biot-GHK (from Sederma), Pal-KMO2K (Sederma) and derivatives thereof. Suitable tetrapeptide derivatives for use in the present invention include, but are not limited to: N-palmitoyl-GQPR (SEQ ID No.: 3) (from Sederma). Suitable pentapeptide derivatives for use hereinIncluding, but not limited to: N-palmitoyl-KTTKS (SEQ ID NO: 2) (Matrixyl available from Sederma corporation)^TMObtained), N-palmitoyl-Tyr-Gly-Phe-X with X Met or Leu (SEQ ID no: 10) or mixtures thereof. Suitable hexapeptide derivatives for use herein include, but are not limited to: N-palmitoyl-VGVAPG (SEQ ID NO: 1) and derivatives thereof. Pal-GHK and Pal-GQPR (SEQ ID NO: 3) (matrix yl)^TM3000, Sederma) mixtures can also be mentioned.

The most preferred commercially available composition containing tripeptides or derivatives includes the Sederma company's bio-peptide-CL^TM，Maxilip^TM，Biobustyl^TM，Procapil^TMAnd Matrixyl^TMAnd synthe' 6^TM. The preferred tetrapeptide-derived compositions available on the market include Rigin offered by Sederma corporation^TM，Eyeliss^TM，Matrixyl^TMLoaded and Matrixyl3000^TM(ii) a They contain between 50 and 500ppm palmitoyl-GQPR (SEQ ID No.: 3) and a carrier.

As additional active ingredients, the following commercially available peptides can also be mentioned: vialox^TM，Syn-ake^TMOr Syn-Coll^TM(Pentapharm, translation: trade name and company name), Hydroxyprolisea CN^TM(Exsymol)，Argireline^TM，Leuphasyl^TM，Aldenine^TM，Trylgen^TM，Eyeseryl^TM，Serilesine^TMOr Decorinyl^TM(Lipotec)，Collaxyl^TMOr quintesine^TM(Vincience)，BONT-L-Peptide^TM(lnfinitec Activos)，Cytokinol^TMLS(Laboratoires Serobiologiques/Cognis)，Kollaren^TM，IP2000^TMOr Meliprene^TM(lnstitut Européen de Biologie Cellulaire)，Neutrazen^TM(Innovations)，ECM-Protect^TM(Atrium Innovations)，Timp-Peptide^TMOr ECMModuline^TM(lnfinitec Activos)。

A)Examples of obtaining albizzia julibrissin extract that can be used according to the present invention

Example 1 of the protocol: thermal extraction of albizzia seeds under reflux is carried out using an alcohol or a mixture of alcohols, preferably ethanol. After removal of the alcohol, the residue was taken up with a glycerol/water mixture.

Alternatively, the extraction can be performed using a mixture of alcohol and water.

Example 2 of the procedure: hot aqueous extraction under reflux of albizzia was performed. The obtained extract is mixed with glycerol to form a water-glycerol mixture.

The water-glycerol albizzia extract thus obtained was titrated in albizzia acid identified by HPLC chromatography. It can be directly used in herbal formulation; or further diluted in excipients to form "active ingredients" for future galenic formulations as explained below.

B)Formula of active ingredients

The active ingredient is a composition comprising the aqueous-glycerolic albizzia extract obtained according to A) above, dissolved in a matrix to form a physiologically acceptable medium. This active ingredient is particularly suitable for the cosmetic industry for the preparation of cosmetic formulations, such as emulsions, gels, etc. (see examples of galenical formulations given below).

For the albizzia extract obtained by titration in 2% albizzia acid according to a) above, an effective amount thereof (i.e. 1% to 15%) can be mixed into all hydrophilic matrices such as gels, aqueous buffer solutions, glycerol or any other polyol with physiologically acceptable short chains.

For example, and to describe in vivo testing and herbal formulations, it is preferred to dissolve the 7.5% water-glycerol extract in glycerol. It is this active ingredient which is self-formulated in cosmetic compositions suitable for application to the skin, the optimum range being 1 to 5%. According to the present invention, the active ingredient may be a composition containing an extract of albizzia julibrissin in combination with siegesbeckia orientalis glycosides.

For the albizzia extract obtained by titration in 2% albizzia acid according to a) above, its effective amount (i.e. 1% to 15%) can be mixed with an effective amount (i.e. 0.01% to 1%) of siegesbeckia glycosides in all hydrophilic matrices; such as a gel, an aqueous buffer solution, glycerol or any other polyol having physiologically acceptable short chains.

As an example for in vivo testing and formulation of herbal preparations, for example, given below, an active ingredient was prepared. It contains 7.5% water-glycerin albizzia julibrissin extract, and is titrated in 2% + 0.05% siegesbeckia orientalis glycosides in butylene glycol matrix (extracted from siegesbeckia orientalis; in powder form). This active ingredient will be self-formulated in an optimal ratio of 1 to 5% in a cosmetic composition suitable for application to the skin.

C)In vitro testing; anti-glycation activity of albizzia julibrissin extract

According to the present invention, the anti-glycation activity of the albizzia julibrissin extract has been shown in the in vitro tests given below.

For the experiments carried out on cell culture, the 2% albizine-containing aqueous-glycerol extract obtained according to example a) above was used, diluted in a concentration of 0.006% in an aqueous vehicle of the cell culture medium type.

1. Improvement of quality of dermal fibers

a. Measurement of carboxymethyl-lysine (CML) in fibroblasts

Principles and protocols: CML (specific types of AGEs) accumulates on dermal proteins synthesized by fibroblasts over a specific time frame. In vitro, on fibroblasts in culture, this phenomenon can be accelerated by glycation stress.

Methylglyoxal (MGO) at 500 μ M was applied to Human Dermal Fibroblasts (HDFs) to stimulate AGE formation. After this saccharification contact at 37 ℃ for 4 days, MGO was removed from the culture by rinsing and albizzia extract was applied for 5 days. Then, the supernatant was removed and CML released from the fibroblasts was measured by the Elisa (enzyme-linked immunosorbent assay). From the cell layer, the cells were counted in parallel. Two independent experiments were performed.

Results: albizzia julibrissin extract did not produce any change in cell number. Treatment with MGO reduced survival by 44% and increased CML by 124% (p)<0.01, compared to the reference).

At the same time, albizzia julibrissin extract reduced CML by 43% (p < 0.05; compared to the reference with MGO). This test highlights the deglycation feature of the extract of the invention, since it reduces CML after the action of the saccharifying agent.

TABLE 1: changes in CML in cultured fibroblasts

Anodic control aminoguanidine 0.03%: -41% CML; (p < 0.01).

b. Reduction of AGEs in explants

Principles and protocols: human abdominal skin explants taken from 34 year old women were placed in contact with the saccharifying agent MGO dissolved in survival medium. After the glycation phase, the skin (n ═ 5/case) received the application of a cream containing albizzia julibrissin extract or a placebo cream (cream according to herbal formulation paragraph example 1). Topical application was performed once daily for 4.5 days. The skin portion was wrapped and cut with a microtome before being labeled with a fluorescent anti-AGE antibody. 10 photographs were taken for each case and analyzed on appropriate software.

Results: the results show that AGE markers are predominantly located in the dermis, rich in one region of stable matrix proteins (especially collagen). Therefore, quantization is performed in this region.

TABLE 2: changes in AGEs in skin explants

AFU ═ arbitrary fluorescence unit

Treatment of explants with MGO followed by placebo cream resulted in a 23% increase in AGE labeling compared to explants that had received the albizia julibrissin extract cream (p < 0.01). This demonstrates the deglycation phenomenon; the presence of albizzia julibrissin extract helps the explants to break free of a part of the formed AGEs (by reference).

c. Reduction of vimentin damage

Principles and protocols: like proteins with a long half-life (such as collagen), proteins with a shorter half-life can also be altered by glycotoxins. Vimentin was studied in this experiment; its role in tissue for cell contraction and dermal adhesion is mentioned above. We have also known that this protein is sensitive, especially to MGO forming CML. Therefore, it is very important to protect it.

Labeling of vimentin was performed with fluorescent anti-vimentin antibody using the same explants as above (point 1 b); the same method is used for quantization.

Results: the photograph shows the vimentin markers in the upper dermis. Therefore, quantization is performed in this region.

TABLE 3: change in vimentin markers on skin explants

UFA (arbitrary fluorescence units)

The beneficial effect of albizzia julibrissin extract against glycation stress clearly appears as if the extract seems to have "deglycated" a significant fraction of the vimentin that has been glycated in the initial stage (by reference).

a. Improvement of shrinkage ability

Principles and protocols: the glycotoxins alter the mechanical properties of dermal collagen and elastin, resulting in dermal disruption and hardening. A gel containing collagen I and III and dermal human fibroblasts was prepared (n ═ 4/case). After polymerization, the gel was treated with MGO ± albizzia extract for 5 days. The shrinkage was monitored using a CCD camera. The difference between the gel surface at days T0 and T5 is a good indicator of gel shrinkage.

Results: MGO treatment of placebo gel reduced its contractility by 78%; this can be attributed to the reduced survival of fibroblasts and the destruction of the gel by the AGEs formed. Albizzia julibrissin extract improved contractility by 68% compared to the reference, if compared to the stress case.

TABLE 4: variation of dermal lattice shrinkage

2. Improvement of respiratory capacity

Improvement of contractile capacity in ATP-deficient situations

Principles and protocols: a gel containing collagen I and III and dermal human fibroblasts was prepared (n ═ 4/case). After polymerization, the gel receives 2DG (2-deoxyglucose), a non-energetic analog of glucose (non-glycation), a reversible inhibitor of ATP formation. These cultures also received albizzia extract, with or without, for a period of 5 days. The shrinkage was monitored using a CCD camera. The difference between the gel surface at T0 and T5 days on this same surface is a good indicator of gel shrinkage.

As a result:the reference gel contracted as expected. For the reference case, treatment with 2DG, which reduced ATP supply, reduced contraction by 75% without changing survival. Albizzia julibrissin extract improved contraction by 98% (p) despite ATP deficiency (using 2DG)<0.05; compared to the reference case). This suggests that the extract allows cells to better maintain their contractility despite the lack of ATP.

TABLE 5: changes in dermal lattice shrinkage following 2DG contact

Improvement of respiratory capacity under "fatigue" stress

Principles and protocols: the following culture protocol is intended to simulate the fatigue and resting phase of cells. HDF was continuously cultured for 16 days with or without albizzia julibrissin extract. On days 4 and 13, cells were subjected to fatigue stress for 3 days using 500 μ M of MGO. ATP was measured at the end of all cases (n ═ 5/case; 2 independent experiments).

As a result:in the absence of saccharification, there was no significant difference between the reference case and the case using albizzia julibrissin extract. In the reference case, treatment with MGO (glycation stress) greatly reduced the amount of ATP (-74%) compared to the non-glycated case. In the case of saccharification in the presence of the albizzia julibrissin extract, ATP synthesis was 182% compared to the control case after saccharification.

Therefore, the albizzia julibrissin extract can maintain a good ATP synthesis level in cells in spite of glycation stress as compared with the reference substance.

TABLE 6: change in ATP concentration

Protection of AMPK energy sensors

Glycotoxins reduce the functional capacity of mitochondria, especially by altering their respiratory enzymes. The glycotoxin also attacks certain intracellular sensors responsible for detecting fluctuations in AMP and ATP levels, such as AMP-dependent kinase (pAMPK); thereby adjusting the strength of cellular respiration as required. Fast and responsive energy production requires that the enzyme cannot be saccharified, keeping its properties intact.

Principles and protocols: human abdominal skin explants taken from 34 year old women were placed in contact with MGO dissolved in survival medium. After the glycation phase, the skin (n-5/case) received application of either albizzia extract or placebo cream. Topical application was performed once daily for 4.5 days. The skin sections were cut with a microtome before labeling with fluorescent anti-pAMPK antibody. 10 photographs were taken for each case and analyzed on appropriate software.

As a result:the results show that the pAMPK marker is significantly located on the dermis where quantification is performed. The topical application of the cream containing the albizzia julibrissin extract allows the cells to protect the energy sensor AMPK better than the reference case. Thus, the extract appears to have been "deglycated" by a portion of the enzymes that had been saccharified at the initial stage.

TABLE 7: changes in pAMPK marker

Concentration of	pAMPK(AFU*)	% change; significance of
			Reference object with MGO	0.87±0.30	Datum
Extracts of MGO and Albizzia julibrissin	1.09±0.40	+25％；p<0.01

AFU ═ arbitrary fluorescence unit

3. Sugar toxin detoxification system

a. Maintenance of glyoxalase-1

Principles and protocols: as can be seen earlier, once they are formed by glyoxalase-1, MGO and GO can be neutralized; since glyoxalase-1 prevents the formation of the final glycotoxin and the amplification of the synthesis of these protein-glycation agents.

500 μ M Methylglyoxal (MGO) was applied to the healed human dermal fibroblasts with the aim of stimulating the formation of glycotoxins in their extracellular matrix. After the saccharification contact period at 37 ℃, MGO was removed from the culture by rinsing and albizzia extract was applied for 5 days. Subsequently, the cell layer was extracted, and the remaining glyoxalase-1 was quantified by Western Blot (Western blotting). The comparison was performed using equivalent amounts of protein (protein assay by BCA).

As a result:there was no significant difference between the reference case and the case using the albizzia julibrissin extract in the absence of the saccharification treatment. In the reference case, the glycation treatment resulted in a 56% decrease in glyoxalase-1 synthesis (p)<0.01, compared to the non-saccharified reference). After further saccharification in the presence of the albizzia julibrissin extract, there was still 41% more glyoxal than the saccharification case referenceAn enzyme. The extract of the invention appears to have "deglycated" an important part of the enzymes that have been saccharified during the initial incubation period.

b. Maintaining proteasome activity

Principles and protocols: proteasome is an intracellular protein structure; cell content decontamination, specifically to regulate damaged proteins, especially by glycation stress, can become toxic. Proteasomes are sensitive to sugar toxins; the process can be direct (via saccharification) or indirect (if ATP synthesis is low, as it is ATP dependent).

Serum Albumin (BSA) was used as a protein model. The BSA was saccharified for one week under exposure to MGO. This glycotoxin (glycated BSA) was then incubated for 6 days with or without albizzia extract.

Each of the two solutions was allowed to contact human dermal fibroblasts at the healed site for 3 days to induce proteasome changes by glycated proteins. After this contact, and after rinsing, the cells were crushed and the proteasome activity assay was performed.

Results: in the absence of saccharification, the albizzia julibrissin extract had no effect on proteasome activity. These results show that model glycotoxin (BSA-AGE) significantly reduced the activity of the cellular purification system (i.e. proteasome) by 16% by its contact with the cells (two independent experiments). This experiment shows that extracellular glycotoxins can regulate effects in cells through different metabolic pathways. The same BSA-AGE complex lost its toxicity after 6 days of contact with albizzia julibrissin extract and became unable to reduce proteasome activity in human dermal fibroblasts. It can be hypothesized that the glycotoxin model (BSA-AGE) was detoxified (deglycated) by 6 days of exposure to albizia julibrissin extract. .

TABLE 8: changes in proteasome Activity following contact with BSA-AGE

Proteasome activity in units of Δ UA/min/10⁶Individual cell

Reduction of AGE-pigmentation

Principles and protocols: MGO is indirectly involved in the production of another group of toxins called lipofuscin. This pigment is seen as yellow on the skin site. This is a poor feature because it is formed from a collection of different damaged cellular components. A reduction in the function of the proteasome will allow it to accumulate. The accumulation of this toxic pigment is promoted by the proteasome saccharification (and thus alteration). The same "fatigue" cell stress as in bar 2a above was used, alternating two rest periods (+/-albizzia extract) and 2 stress periods (+/-albizzia extract, using 500 μ M MGO). At the end of the second glycation stress, a dose of lipofuscin marker was performed on the cells and reduced to the number of cells.

As a result:the results show that MGO produced a large increase in dityrosine accumulation (+ 129%, p)<0.01). At the same time, albizzia julibrissin extract significantly limits its formation.

Albizzia julibrissin extract has the ability to substantially reduce dityrosine formation (for this case, undetectable in our experiments); in the presence of the extract, the amount of dityrosine observed with MGO was lower than that of the negative reference (no MGO).

TABLE 9: changes in lipofuscin AGE-pigment accumulation

AFU: an arbitrary fluorescent unit; positive MGO reference: tocopherol 500 μ M: -61.1% (p <0.01)

Reduction of effects of ages on angiogenesis

Principles and protocols: AGEs are formed in the direct environment of blood vessels, or in the blood vessels themselves; this can result in the accumulation of waste and fluids, resulting in a dark color under the eye. Het-CAM uses eggs that are very early in their development and the nervous system has not yet fully developed. The inventors developed an original method for assessing changes in AGE-related angiogenesis.

BSA was saccharified with MGO. At the end of this period, the glycotoxin (glycated BSA) was left in contact with the albizzia julibrissin extract or not for the period of one day.

Then, the two types of saccharification solutions are applied to the egg membranes. As a result of AGEs-induced vasodilation, blood vessels that have previously become invisible at this point now become clearly visible. The results were scored to assess the reduction in negative effects on the blood vessels.

Results

Watch 10: reduction of negative AGE-related effects of Albizzia julibrissin extract in Het-CAM assay

After applying BSA-AGE to the egg vascular network, its toxic effects manifest rapidly and severe damage can be seen, with concomitant leakage and development of vascular networks.

After application of BSA detoxified (deglycated) with albizia julibrissin extract, a large, dose-dependent reduction in damage resulted: 0.075% of the extract (p <0.01) is-55%; 0.225% of the extract (p <0.01) is-70%. Thus, albizzia julibrissin extract helps protect the vascular network, prevent deleterious damage by the glycotoxins, reduce blood leakage and excessive blood vessel formation, thereby reducing the color of the dark circles of the eye, as compared to the reference.

e. Protection of fibroblast melatonin synthesis

Principles and protocols: as previously mentioned, melatonin often emphasizes its cytoprotective benefits. Changes in Human Dermal Fibroblast (HDF) melatonin synthesis were monitored in the presence or absence of Methylglyoxal (MGO). Accordingly, a dose dependent decrease in melatonin due to this stress of up to-20% after 2 hours of exposure is demonstrated.

In a second series of experiments, HDF had been in contact with the extract of the invention for 24 hours; next, after rinsing, the MGO has been contacted as before. After 24 hours, melatonin synthesis was monitored.

Results

TABLE 11: changes in melatonin synthesis caused by glycation stress; effect of Albizzia julibrissin extract of the present invention

MGO resulted in a 32% reduction in melatonin (p <0.05) compared to the reference; the extract of the invention used in the protection maintains melatonin at a steady state concentration, preventing its loss.

D)In vitro testing: albizzia julibrissin extract and siegesbeckia orientalis glycoside combined activity

1.3 expression of senescence genes

The term "senescence genes" relates to all genes representing their changes in activity during the aging process, or all genes that play a role in maintaining homeostasis in the interest of their function.

a) FOXOs (forkhead transcription factor O subfamily)

The O subfamily of the forkhead transcription factor means the regulation of oxidative damage. When deacetylated, FOXOs proteins are more active. Cellular stress results in their acetylation and reduced activity. Sirtuins (SIRT1) are one of the major activators of FOXOs pathway. The FOXO3a gene disappearance in human dermal fibroblasts has similarities to the senescence phenotype: the greater generation of ROS (reactive oxygen species) and the doubling time of the cell population are important.

b) Stress-inducing protein (Sesns)

Stress-inducing proteins are stress-responsive proteins. They are able to inhibit oxidative damage by allowing the elimination of defective mitochondria (ROS-producing bodies) by direct action similar to antioxidants, or similar to autophagy inducers. Therefore, they are considered as physiological brakes in the aging process.

The effect of the composition of the invention was tested using the SIPS (stress induced premature aging) protocol.

Human dermal fibroblasts were cultured for 3 days. They then received or did not receive the product of the invention (test article) and were tested at different concentrations for an additional 3 days. Cells were then subjected to oxidative stress stimulation (H2O2) within 2 hours. . The cell monolayer was again contacted or not contacted with the test product for 6 hours. Next, RNA was provided and the expression of 3 senescence genes was quantified by real-time RT-PCR.

Cells treated with the composition of the invention were compared to untreated cells.

TABLE 12：

For the reference, the ratio is 1

The above results show that significant induction of this senescence gene occurs with the composition of the invention compared to untreated cells.

2. Non-enzymatic saccharification of elastin

The non-enzymatic saccharification that occurs between proteins and reducing sugars (e.g., fructose or glucose) is a spontaneous, slow reaction that can be accelerated by heat. In this test, the protein was soluble bovine elastin; the reducing sugar is fructose. They were incubated for 9 days at a temperature of 50 ℃ with or without the test product. The end product resulting from saccharification can be quantified by fluorescence (λ ex-360 nm, λ em-460 nm).

Watch 13：

Aminoguanidine 0.03%: -99%; p <0.01

The above results show that Albizzia julibrissin extract resulted in a decrease in nonenzymatic glycation of elastin (-23%; p <0.05) compared to the reference. Although siegesbeckia glycosides alone had no effect, the combination of albizia julibrissin extract with siegesbeckia glycosides showed a synergistic effect (-31%; p < 0.01).

3. Inhibitory Activity of metalloprotease MMP1 and MMP2

Human dermal fibroblasts were cultured until confluence. The cells were then left with or without contact with the test product for 24 hours. Extracts of cigarette smoke are then used to stimulate cells with or without the test product. After the last incubation with or without test product, the metalloproteinases MMP1 and MMP2 present in the culture medium were measured by ELISA and reduced to cell number (assessed using Hoescht reagent).

TABLE 14：

These results show an increase in MMPs after stress. The combination of the invention results in a reduction of stress-induced MMPs in treated cells compared to reference cells.

4. Increase of collagen I in human dermal fibroblasts

Human dermal fibroblasts were cultured for 24 hours. The cells were then left with or without test product for 6 days. Finally, the collagen I produced by the cells is quantified by fixing the immune markers of the cell layer.

Watch 15：

The combination of the invention stimulates collagen I synthesis in treated cells compared to the reference cells.

E)Examples of galenic formulations

The active ingredients of the invention: a composition comprising an extract of albizzia julibrissin prepared according to paragraph B.

Various formulations, with or without additional cosmetic active ingredients, will now be described; for each case, the latter acts to support and/or supplement the activity of the active ingredients of the invention. These ingredients may be of any kind, depending on their function, application site (body, face, neck, chest, hands, etc.), desired end effect and target consumer.

Example 1: cream form

Protocol: weighing phase A; it was allowed to soak for 30 minutes without stirring. Heating phase A in 75 deg.C water bath. Weigh and mix phase B. Weigh phase C, heat to 75 ℃ in a water bath and mix thoroughly. Add phase B to phase a. And (4) uniformly mixing. Pouring the phase C into the phase A + the phase B under stirring. Mix thoroughly. Add phase D without preparation. Add phase E and mix thoroughly. Add phase F and mix thoroughly.

Example 2: late paste form

Protocol: phase A was weighed and heated in a water bath at 85 ℃. Weighing phase B, and heating in water bath at 85 deg.C. And weighing the phase C. Before forming the emulsion, add phase C to phase A and mix thoroughly. The A + C phase was slowly poured into the B phase with vigorous stirring. Thoroughly mixed with stirring for cooling. Add phase D to the emulsion at about 35 ℃ and mix thoroughly.

Can be added to the formulaAdditional ingredient examples：

Covi-Ox T90^TM: antioxidant actives (tocopherols) sold by Cognis corporation. For example, 0.1% of it can be added to the B phase.

Crodarom Goji^TM: antioxidant, detoxifying and humectant actives sold by Crodarom corporation. It may be added to phase a at the same time as C, e.g. up to 2.50%.

Example 3: post-emulsion form (gel-cream)

Protocol: phase A was weighed and placed under propeller stirring. Weigh and mix phase B. Phase B was added to phase a with stirring and homogenized thoroughly for 30 minutes. The A + B phases were heated in a water bath at 75 ℃. Phase C was weighed and heated to 75 ℃ in a water bath. Phase C was poured into phase A + B with Staro stirring and mixed thoroughly. Weigh and homogenize phase D. At around 40 ℃, add phase D to the previous phase and mix thoroughly. The pH was adjusted to 5.00-5.50 using phase E below 35 ℃ and mixed thoroughly. Add phase F and mix well. Add phase G and mix well. Checking the pH value to be 5.50-5.00.

Can be added to the formulaAdditional ingredient examples：

Kelisoft^TM: anti-hair-regrowth actives marketed by Sederma corporation. It can be added at the end of the formulation process, up to 3.00% before the G phase.

Calmosensine^TM: sensitive skin soothing actives sold by Sederma corporation. It can be added at the end of the formulation process, up to 3.00% before the G phase.

NG Seve de Bouleau^TM: tinting and moisturizing agents sold by Sederma corporation. It can be added at the end of the formulation process, up to 3.00% before the G phase.

Example 4: gel form for eye contouring

Protocol: phase a was weighed and homogenized. And weighing the phase B, and mixing uniformly. Phase B was added to phase a with propeller agitation. It was allowed to soak. Weighing phase C, and homogenizing. Phase C was added to phase a + B under vigorous helical stirring. It was homogenized for 1 hour under ordinary propeller agitation. The pH was adjusted to 5.50 using phase D. Homogenisation was carried out with propeller agitation. Add phase E to the previous phase and mix thoroughly. Add phase F and mix thoroughly.

Can be added to the formulaAdditional ingredient examples：

EYELISS^TM: sederma corporation sells eye contour actives (particularly for dark circles and bags). 3.00% may be added to phase E.

Example 5: mask form

Protocol: phase A was weighed and placed under propeller stirring. Weigh and mix phase B. Phase B was added slowly to phase a with propeller agitation and mixed thoroughly. The A + B phases were heated in a water bath at 80 ℃ for 2 hours without stirring. Stirred rapidly and then allowed to cool under stirring. Phase C was added to the previous phase at around 40 ℃. The pH was adjusted to 5.50 with phase D below 35 ℃ and mixed thoroughly. Phase E was added with slow stirring. Add phase F and mix thoroughly. Add phase G and mix thoroughly. Checking the pH value to be 5.0-5.5.

Can be added to the formulaAdditional ingredient examples：

KOMBUCHKA^TM: sederma corporation sells actives that act on the luminosity of a complexion. 3.00% can be added at the end of the formulation, before phase F.

Novaplant Ginger Special^TM: crodarom company sells active substances that increase blood circulation. 2.00% diluted in Crosupply CAB 30 (Cocamidopyl Betaine) can be added after phase D.

Example 6: gel form for face

Protocol: phase a was dispersed with stirring. Ultrez 10 was sprinkled into water and allowed to soak for 30 minutes. Heat phase C until completely dissolved. Mixing the phase A and the phase B. Add phase C to phase (B + a). Phase D was added to phase (a + B + C) with stirring. Add phase E. Neutralizing with F phase. Add phase G and mix.

Can be added to the formulaAdditional ingredient examples：

RIGIN^TM: the actives sold by Sederma corporation improve the elasticity and firmness of the skin and increase the hydration and smoothness of the skin. For example, 3% by weight may be added to the G phase.

MATRIXYL 3000^TM: peptide-based anti-wrinkle formulations sold by Sederma corporation; it helps to repair skin damage caused by aging. For example, 3% by weight may be added to the G phase.

MATRIXYL synthe’6^TM: peptide-based anti-wrinkle formulations sold by Sederma corporation; it helps to repair skin damage caused by aging.For example, 2% by weight may be added to the G phase.

Example 7: other cream forms

Protocol: phase A was weighed and allowed to soak for 30 minutes. Then, phase A was heated in a water bath at 75 ℃. Heat phase B until dissolved. Add phase B to phase a. Phase C was heated in a water bath at 75 ℃. Phase C was added to phase (a + B) with stirring. The phases were thoroughly stirred. Neutralization was carried out at about 55 ℃ by means of the E phase. Phase F, then phase G, was added and homogenized thoroughly.

Can be added to the formulaAdditional ingredient examples：

DERMAXYL^TM: anti-aging actives sold by Sederma corporation; it can smooth wrinkles and repair skin barrier. For example, 2% by weight may be added to the (A + B + C) phase.

Retinol, Resveratro et Niacinamide: anti-aging ingredients, especially anti-wrinkle agents. For example, 0.1% rosin oil or 0.5% resveratrol may be added to the (a + B + C) phase. For example, a 10% aqueous solution of nicotinamide may be added to phase F.

CHRONODYN^TM: actives sold by Sederma corporation; it is beneficial to toning and firming the skin and removing fatigue signs. For example, 3% may be added to the F phase.

VENUCEANE^TM: actives sold by Sederma corporation; it can prevent visible signs of photoaging (spots, wrinkles, dryness), protect cellular structures, prevent damage from ultraviolet light, and enhance skin strength. For example, 3% may be added toAnd F phase.

Example 8: cream form for body

Protocol: phase A was weighed and allowed to soak for 30 minutes. Phase A was heated in a water bath at 75 ℃. Heat phase B until dissolved. Add phase B to phase a. Phase C was heated in a water bath at 75 ℃. Phase C was added to phase (a + B) with stirring. Add phase D and mix thoroughly. Neutralizing with E phase at about 55 deg.C, and mixing. Add phase F and phase G and mix thoroughly.

Can be added to the formulaAdditional ingredient examples：

JUVINITY^TM: actives sold by Sederma corporation; it reduces the signs of aging on the face and "cleavage", smoothes wrinkles, restructures and densifies the dermis. For example, 2% may be added to the (a + B + C) phase.

Tocopherol (vitamin E) or uAcid (ALA): active substance with antioxidant and anti-free radical properties. For example, 0.5% by weight of the (A + B + C) phase may be added.

O.D.A.white^TM: actives sold by Sederma corporation; by reducing melanin synthesis, it can lighten the skin. For example, 1% may be added to the (a + B + C) phase.

Bio-Bustyl^TM: actives sold by Sederma corporation; containing peptides and bacterial filtrate, has a robust and toning global use for female breasts. For example, 3% can be added to the F phase.

Example 9: in the form of a slurry

Protocol: phase A: the carbomer was sprinkled in water and allowed to stand for 15 minutes. Mixing the phase B. Pouring phase B into phase A, and homogenizing. Phase C was weighed, mixed and added to phase a + B with stirring. It was allowed to soak for 1 hour. Without preparation, phase D is added to the previous phase under stirring. Neutralizing with E phase. Stirring was started. Then phase F is added. Allow to mix for at least 1 hour with stirring, then add phase G. And (4) uniformly mixing.

Can be added to the formulaAdditional ingredient examples：

LUMISPHERE^TM: actives sold by Sederma corporation; it is diacetyl boldine (DAB) and modified manganese titanium dioxide (TiO) which are arranged in polymethyl methacrylate micro-envelope₂Mn). TiO 2₂Mn can give uniform and bright luster effect to skin; whereas DAB provides a physiological effect of highlighting. For example, 4% may be added to the F phase.

REVIDRATE^TM: actives sold by Sederma corporation; it is particularly effective in improving the cohesion and hydration of the epidermis. For example, 2% may be added to phase C.

EVERMAT^TM: actives sold by Sederma corporation; it can reduce sebum secretion, and thus is involved in the treatment of oily skin. For example, 4% may be added to the F phase.

HALOXYL^TM: actives sold by Sederma corporation; by resorbing the eye circles, it improves the eye contour. For example, 3% may be added to the F phase.

Example 10: lotion form

Protocol: weighing phase A. Weighing phase B, and mixing. Phase B was added to phase A over a period of 30 minutes with stirring. Weigh phase C and mix until a homogeneous blend is obtained. Phase C was added to phase a + B with stirring. Add phase D to the previous phase. Add phase E under stirring and mix well. Phase F was weighed, mixed and added to the previous phase and homogenized thoroughly.

Can be added to the formulaAdditional ingredient examples：

EYELISS^TM: as described above. For example, 3.00% can be added to phase E.

Ac-Net^TM: actives sold by Sederma corporation; has the ability to complete the treatment of oily and acne-prone skin. For example, 3% may be added to the E phase.

EVERMAT^TM: as described above. For example, 4% may be added to the E phase.

HYDRERGY^TM: actives sold by Sederma corporation; it is a long-term humectant and stimulates the synthesis of ATP. For example, 3% may be added to the E phase.

Example 11: capillary washing liquid

Ingredients	INCI name	By weight%
			Phase A
Incroquat CTC 30	Cetroronium chloride	1.00
			Citric acid	Citric acid	0.22
Trisodium citrate	Trisodium citrate	1.20
			Sorbic acid ester	Potassium sorbate	0.10
H2O		Qsp
			Phase B
Nipagine	Nipagin methyl ester	0.20
			Procetyl AWS	PPG 5Ceteth 20	2.00
Phase C
			Active ingredients of the invention		3.00
Phase D
			Crillet 1	Polybehenyl ester 20	1.00
Perfume	Perfume	0.10

Example 12: lip balm

Protocol: phase A was heated at 85 ℃. Mix phase B and heat at 85 ℃. And (4) mixing. Slowly pour phase a into phase B with stirring (Staro s 3000, then s 1200). Add phase C preheated at 80 ℃ and homogenize. Phase D is added at about 35 ℃. And (5) casting.

F)Special examples of galenical formulations of the combination of albizzia julibrissin extract and siegesbeckia orientalis glycoside according to the invention

The active ingredients of the invention: a composition comprising albizzia julibrissin extract and siegesbeckia orientalis glycoside prepared according to paragraph B.

Various formulations, with or without additional cosmetic active ingredients, will now be described; for each case, the latter acts to support and/or supplement the activity of the active ingredients of the invention. These ingredients may be of any kind, depending on their function, application site (body, face, neck, chest, hands, etc.), desired end effect and target consumer.

Example 13: cream form, especially for the contour of the eye

Protocol：

Without stirring, disperse phase A in water, and stand for 1 hour. Homogenize phase B and add it to phase A over 30 minutes under propeller agitation. Phase C was added to phase A + B with stirring of taro. Without preparation, phase D was added to the previous phase under stirring of taro. Neutralization was carried out with phase E under stirring of taro. Phase F is mixed and added to the previous phase with stirring at taro. Adding the G phase into the former phase, and mixing uniformly.

Can be added to the formulaAdditional ingredient examples：

MATRIXYL 3000^TM: peptide-based anti-wrinkle formulations sold by Sederma corporation; it can help repairSkin damage due to aging. For example, 3% by weight may be added at the end of the formulation.

RESISTEM^TM: anti-aging ingredients sold by Sederma corporation; it is a plant extract obtained by stem cell culture of heart leaf corms. It helps to reduce the levels of pro-ageing agents and topical micro-inflammation, reduce skin redness, and enhance natural skin glow. For example, 3% by weight may be added at the end of the formulation.

Example 14: gel form, especially for eye contours

Procedure:without stirring, disperse phase A in water, and stand for 1 hour. Phase B was dispersed in water under rapid propeller stirring and allowed to mix for 30 minutes. Phase C was weighed and homogenized. Phase C was added to phase B with propeller agitation. And standing for 30 minutes. Add phase a to phase B + C with stirring by the blade. Phase D was added to the previous phase with propeller agitation followed by blade agitation. The previous phase was neutralized with phase E under agitation by a blade. Adding the G phase into the previous phase, and uniformly mixing.

Can be added to the formulaAdditional ingredient examples：

IDEALIFT^TM: lipid-based formulations sold by Sederma corporation; it can resist skin sagging and improve resistance to gravity. 4% can be added at the end of the formulation.

Example 15: in the form of a slurry

Procedure:phase A was weighed and placed under propeller stirring. Weigh and homogenize phase B. Add phase B to phase a and mix well. Weighing phase C, and mixing. Phase C was added to phase A + B with propeller agitation. Without preparation, phase D was added with propeller agitation. Phase E was added and held for 30 minutes with stirring by the blade to homogenize.

Can be added to the formulaAdditional ingredient examples：

RIGIN^TM: the actives sold by Sederma corporation improve the elasticity and firmness of the skin and increase the hydration and smoothness of the skin. 3% by weight may be added at the end of the formulation.

Example 16: makeup removing emulsion

Ingredients	INCI name	By weight%
			Phase A
H2O	Water (W)	Qsp100
			Potassium sorbate	Potassium sorbate	0.10
Phase B
			Butanediol	Butanediol	35.00
Crovol A70-LQ-(RB)	PEG-60 Almond glyceride	1.00
			Crodateric CAB 30-LQ-(RB)	Cocoamidopropyl betaine	1.00
Tween 20-LQ-(RB)	Polybehenyl ester-20	2.00
			Phenoxyethanol	Phenoxyethanol	Qs
Perfume	Perfume	0.10
			Phase C
The ingredients of the invention		3.00

Procedure:phase A was weighed and placed under gentle propeller stirring. Weigh and homogenize phase B. Phase B was added to phase A with rapid propeller stirring. Add phase C to phase a + B with gentle propeller agitation. Phase D was added with gentle propeller stirring.

Can be added to the formulaAdditional ingredient examples：

Birch SAP^TM: skin toning and moisturizing formulations sold by Sederma corporation are based on stock solutions from birch sapwood. 3% may be added at the end of the formulation.

Example 17: eye contour Patch (for processing Black eye and under-eye bags)

Ingredients	INCI name	By weight%
			Phase A
H2O	Water (W)	Qsp100
			Potassium sorbate	Potassium sorbate	0.10
Phase B
			Glycerol	Glycerol	3.00
Phenoxyethanol	Phenoxyethanol	qs
			Satiagel VPC 614	Carrageen gum extract	1.00
Viscogum BCR 13/250	Locust bean gum and sucrose	1.00
			Phase C
H2O	Water (W)	5.00
			NaOH 30％	Potassium hydroxide	0.50
Phase D
			The ingredients according to the invention		3.00
Phase E
			Perfume	Perfume	0.10

Procedure:phase A was weighed and placed under propeller stirring in a water bath at 85 ℃. Weigh and homogenize phase B. Phase B was added to phase A with propeller agitation. It was left to homogenize for 1 hour with stirring by the blade. It was removed from the water bath and allowed to cool with agitation by a blade. At below 70 deg.C, phase C is added. Adding phase D. Add phase E. The patch was poured into a mold and allowed to cool at room temperature for 2 hours.

Can be added to the formulaAdditional ingredient examples：

Wonderlight^TM: brightener sold by Sederma corporation. Before cooling, 3% can be added to the formulation process before phase C.

Example 18: fluid gel form

Ingredients	INCI name	By weight%
			Phase A
H2O	Water (W)	Qsp100
			Potassium sorbate	Potassium sorbate	0.10
Phase B
			Renex G26	Glycerol polyether-26	5.00
Phenoxyethanol	Phenoxyethanol	Qs
			Crovol A70	PEG-60 Almond glyceride	0.50
N-Hance HP40	Hydroxypropyl guar gum	0.40
			Keltrol CG-SFT	Xanthan gum	0.50
Phase C
			Tween 20	Polybehenyl ester-20	1.00
Perfume	Perfume	0.10
			Phase D
The ingredients according to the invention		3.00

Procedure:phase A was weighed and placed under propeller stirring. Weigh and homogenize phase B. Phase B was added to phase A with propeller agitation. And (4) uniformly mixing. Weigh phase C and homogenize. Phase C was added to phase A + B with propeller agitation. And (4) uniformly mixing. Adding the phase D into the previous phase, and uniformly mixing.

Examples of additional ingredients that can be added to the present formulation:

Skin Tightener^TM: the ingredients sold by Sederma corporation, the union of prolamin and polymannuronic acid; it can brighten and smooth. 10% can be added at the end of the formulation.

G)In vivo evaluation of saccharification activity of Albizzia julibrissin extract

In the in vivo tests given below, the saccharification activity of albizzia julibrissin extract according to the present invention has been shown.

A cream prepared according to example 1 was used for these tests.

Principle of

Pharmacodynamic tests were performed on a group of 24 volunteers; these people present with visible signs of stress (signs of fatigue), or net advanced glycation skin levels, on their faces.

In addition, the amount of advanced glycation end products (AGEs) was assessed for a group of 14 volunteers by measurement using the gel strip method. This situation has been sought to be improved for a second group of 20 volunteers with the same characteristics.

In this study, several complementary methods were used:

use of AGE-Reader as described below^TMDevice byTheir in vivo measurement of autofluorescence, and analysis of the content of advanced glycation end products.

AGE content in the gel strip obtained from the forearm (in vitro method) was measured.

Use ofSkin fatigue and relaxation analysis of (1).

Self-completed assessments of facial fatigue recorded by volunteers while awake-such as-once daily and once weekly.

Protocol

Inclusion criteria for specific study

In selecting women in the "stimulated" skin study, AGE-based is the primary-they must be between 50 and 70 years-and, according to AGE-Reader^TMMeasured, showed sufficiently high levels of glycated product.

For signs of fatigue and gel strip samples, the volunteers must have signs of facial fatigue (dark eye circles, facial features of facial distortion, dull complexion) and show excessively relaxed forearm skin. The volunteer must be an active person; they were reported not to have enough time to rest. They must follow a 15 day washout period (placebo only).

In addition, all 3 trials required that the volunteers be kept in a constant hormonal state (no change in contraception or replacement or curative treatment) for 3 months prior to and during the trial.

During the study they had to use only the cosmetics provided.

Study type and duration

The study was performed in a single blind fashion with non-invasive measurements for:

24 volunteers with irritated skin (average age 60 years [ range 53 to 69 years ]); they applied day and night creams made according to the present invention on their main forearms. The placebo cream was then applied to the contralateral forearm.

14 volunteers with signs of fatigue (mean age 43.5 years [ range 38 to 50 years ]); they applied daily and night creams according to the invention on one of their forearms, respectively. The placebo cream was then applied to the contralateral arm.

20 volunteers with signs of fatigue (mean age 43 years [ range 31 to 50 years ]); they applied daily and night creams according to the invention on their face and one forearm, respectively.

The day cream and the night cream according to the invention are applied once a day for 2 months in a massaging manner; the placebo cream was applied twice a day.

The studies are summarized in the following figures.

Statistical studies were performed using student's t-test or Wilcoxon non-parametric test (if necessary). In each case, a two-tailed test on the paired series was used.

Tolerance to stress

Clinical studies are performed under medical surveillance; the results show that the volunteers were very well tolerated by the product.

1. ^TMAnalysis of the content of the end product of the late stage saccharification Using AGE-Reader

Principle of

AGE-Reader^TM(DiagnOptics Technologies Inc.) is the first medical device to allow non-invasive measurement of AGEs accumulation. The skin of the volunteer was exposed to light having a wavelength of 370 ± 50 nm; it activates functional fluorophores, especially AGEs. The reflected light selectively propagates through a 50 μm diameter fiber, measured by a spectrophotometer. With the aid of this device, it is possible to determine the correlation between the autofluorescence of the volunteer skin and their different AGE contents.

In practice, the volunteer places his/her forearm on the instrument, passing 1cm²The window of (2) is measured.

Results

TABLE 16: after application of the cream or placebo cream according to the invention, by equipping with an AGE-Reader^TMAF of (2) measured change in content of glycated product

AF (arbitrary autofluorescence unit), 24 volunteers, and n (3) measurements

The results show that a significant difference in autofluorescence was found after 2 months of daily cream followed by night cream application once daily compared to placebo.

This reduction was-8.6%, reflecting the reduction in the content of the saccharified end product. It is very clear that a large proportion of the volunteers responded (71%).

2.AGEs in research gel strip (in vitro research)

Principle of

Use of AGE-Reader was confirmed using a test on volunteers (N ═ 14) with visible signs of skin fatigue^TMAs seen, there was a decrease in AGEs on cultured cells and in explants treated with albizzia julibrissin extract according to the invention (see in vitro section).

The AGE content on the gel strips obtained two months before and two months after the day cream and the night cream of the present invention were applied to the front arms, respectively, was measured.

The gel strip was removed and stored at-80 ℃. Proteins on the gel strip were extracted into buffer solution, and then the homogenate was loaded onto polyacrylamide gel to run the proteins and transfer them onto nitrocellulose membrane for development by Western Blot. Again, the anti-AGE antibodies already described in the in vitro part are used. Spots were visualized by chemiluminescence over a major band of approximately 60kDa representing keratin. Total protein was measured in parallel with the normalized results.

Results

TABLE 17: change in AGE protein content (keratin) after application of the cream according to the invention (N ═ 14 volunteers).

The results show that AGEs were increased between T0 and T2; moreover, the amount of this increase was reduced by 27% due to the use of the cream containing albizzia julibrissin extract according to the invention.

3.By using Principle analysis of skin fatigue and relaxation

RV 600(Courage&Khazaka corporation), the travel time of a sound wave in the skin after the wave has been emitted from its surface: this indicator is called the resonance run time (RRT for short). The skin probe of the instrument included an acoustic transmitter at a distance of 2mm from the receiver. Since the amount of sound waves propagating in the skin cannot exceed 0.5mm,it allows to detect the quality of the dermis. The speed of sound propagation in a material depends on the density and tension of the material. The acoustic wave first traces the "line" formed by the dermal fibers. Thus, the conditions of these fibers directly affect the propagation of the wave.

By rotating the probe stepwise about its axis, different travel times can be mapped. Fatigued skin has a lower density, less taut fibers, and thus increased travel time. In addition, a skin having fatigue with a poor skin color also has creases, accompanied by preferential directions (skin anisotropy). This changes the propagation time along certain measurement axes.

By usingSeveral parameters obtained by RV 600 can characterize the skin condition. From these parameters, the following parameters were used:

maximum resonance run time (abbreviated RRTmax); it reflects the "slowness" of sound in the skin, and thus skin fatigue.

TRRTmax/RRTmin ratio; it exhibits the anisotropic nature of the skin, with the magnitude of the ratio reflecting the "relaxation" of the skin which is not uniform in all directions.

Results

Watch 18: changes in the retardation (skin fatigue) and relaxation after application of the cream according to the invention

20 volunteers s, n-3 replicates

The results show that with the cream according to the invention, the retardation of sound transmission in volunteers is reduced; the reduction was 26.3% from month 1 and 30.7% from month 2.

At the same time, the relaxation, measured according to the ratio RRTmax/RRTmin, decreased by 40.2% from month T1, compared to T0; an improvement (-44%) was found after 2 months.

After application of the cream according to the invention, the skin thus regains its vitality, which can be shown by a reduction in retardation and relaxation. By usingThese results show that the cream containing the albizzia julibrissin extract of the present invention significantly improves the density and uniformity of dermal fibers.

4.Self-assessment of facial fatigue

Principle of

To obtain information about the "original" state of facial fatigue, each volunteer, after waking up, before the mirror, evaluated different fatigue criteria in real life situations. Various fatigue criteria and study designs are shown in the following figures.

Results

a.General fatigue

Watch 19: reduction of facial fatigue during the application of the cream according to the invention (20 volunteers)

Self-assessment of volunteers' total facial fatigue every two days revealed a constant regular decrease in total fatigue by applying the cream of the invention.

The average score for 20 volunteers ranged from 2.05 (representing moderate facial fatigue) to 1.15 (representing mild facial fatigue).

This indicates that the overall fatigue impression decreased by 36% at month T1 and by 50% after 2 months. This drop became very significant starting on day 10 (p < 0.01).

b.Specific index

In the questionnaire, the volunteers also filled out some details about dark circles, bags under the eyes, facial features of facial distortion and facial darkness. The results of weekly self-assessments performed by volunteers fully confirmed the significant reduction in overall facial fatigue impression. The overall intensity of the four indicators tested dropped from month T1, with a significant additional drop seen at month T2.

Watch 20: by using the cream of the invention, there was a reduction in the early morning signs of facial fatigue (20 volunteers).

The "bags under the eyes" index dropped from 30 to 24 (T1 months, -20%) and then to 19 (T2 months, -37%).

The "black eye" index dropped from 33 to 29 (T1 months, -12%) and then to 20 (T2 months, -40%).

The "facial characteristics of facial distortion" index dropped from 37 to 29 points (T1 months, -22%) and then to 18 points (T2 months, -51%).

The "dull complexion" index decreased from 22 to 16 points (T2 months, -27%).

After 2 months of treatment, volunteers reported that their target signs of fatigue became mild, or absent.

H)In vivo evaluation of saccharification activity of albizzia julibrissin extract and siegesbeckia orientalis glycoside composition

Three separate clinical trials were performed to demonstrate the effect of the product on:

1. wrinkles and eyelids

2. Black eye

Eye look (Eye appearance)

For these tests, the cream of example 13 of paragraph F) was used.

1.Test on wrinkles and eyelids

Principle of

The purpose of this study was to demonstrate the reduction of wrinkles and eyelid folds after two days of application for 56 days.

The study group consisted of 22 white women, with an average age of 60 years (40-79), with crow's feet having crow's feet with crow's feet. The study was conducted in a single blind fashion compared to placebo. Volunteers applied the cream according to the invention on the contour of the eyes twice a day for 56 days. The placebo cream was applied contralaterally.

Different approaches were associated with each study (T0, T56 j).

·Silflo^TMFingerprint for quantification of crow's feet.

FOITS technique for quantification of eyelid folds.

Photo bench for expert panel quantification of crow's feet and eyelid folds.

Results

a. ^TMSilflo fingerprints for quantification of crow's feet

At each measurement time, Silflo was used^TMSilicone Polymer, at selected sites (crow's feet), making a negative impression of the skin. Then, through Quantirides Station^TM(Monaderm corporation), each mold was scanned using the shadow technique. For simplicity, the negative of the fishtail line is illuminated with tangential light (LED lamp; 35). It creates a shadow behind each wrinkle. Image acquisition was performed using a high resolution digital camera (1920x1440 pixels). The image was recorded using 256 gray levels and then passed through a special Mount maps^TMThe software analyzes to quantify its mitigation.

TABLE 21：

After 2 months of application of the cream according to the invention, a very significant improvement (p <0.01) in the main wrinkle volume (-14%), the mean depth (-9.5%), and the wrinkle opening (+ 11.2%) was noted. These improvements are also significantly different (p <0.05) from the minimal changes that occur with the placebo cream.

b.FOITS technique for quantification of eyelid folds

Principle of

FOITS (fast optical in vivo imaging system) is based on the analysis of fringe projections of the area under investigation, here the eyelid. The device used is a Dermatop consisting of a projector and a camera^TM(EOTech Co.). The two instruments are integrated to form a specific angle for triangulation. The investigation region mitigates the resulting deformed fringes, allowing a 3-dimensional reconstruction of the mitigation.

Then, 3D Moutains Map of collection analysis is used^TMSoftware, extracting the contour of the eyelid fold. The height of the fold is calculated, as well as its surface area.

Results

TABLE 22：

After 2 months of application of the cream according to the invention, a significant reduction of 20% of the wrinkle height was noted (p < 0.01). This reduction is also significantly different (p <0.05) with a slight change of almost-1% observed on the sites receiving placebo.

With respect to the descending surface, there was a significant reduction of 13% (p <0.05) after 2 months of application of the cream according to the invention. On the placebo side, there was no change. The difference between the product and placebo was not significant, but the trend was in the range of p-0.16.

c.Photo bench for quantification of crow's feet and eyelid folds by expert panels

Principle of

Through a photographic workbench Headscan including a high definition Nikon D70 digital camera, special lighting and volunteer restraint system^TM(Orion concept corporation), which realizes the standardization of photographs. The volunteer's posture, photograph, and lighting settings are standardized and controlled to be reproducible at a later time. For this evaluation, the photographs were made in diffuse light to see the color and mitigation correctly, with as close to realistic results as possible.

The areas around the eyes (eye contours) taken at T0 and T56j were presented to six experts together; they were asked to say if they agreed to the following statement:

1. eyelid wrinkles are less pronounced.

2. The skin appeared smooth (fine lines and wrinkles reduced).

Results

The problem "wrinkles are less noticeable, skin is smoother? The results of "show that a response of 47% gives a beneficial effect of the product (p <01) relative to a detrimental effect of 10.5%. In addition, comparison with placebo shows that these 47% favorable responses are significantly better than the 28% favorable response obtained with placebo.

Problem "does the skin appear to be smoother (wrinkles and fine lines are reduced)? "show that the 49% response shows the beneficial effect of the product (p <01) versus just 4% adverse effect. In addition, comparison with placebo shows that these 49% favorable responses are significantly better than the 30% favorable responses obtained with placebo.

·

2.Test on dark circles

Principle of

The purpose of this study was to demonstrate the reduction of dark circles twice a day, after 28 days and 56 days of application.

The study group was 24 white women, the average age 18 years, with dark circles under the eyes. The study was conducted in a single blind fashion compared to placebo. Volunteers applied the cream according to the invention on the contour of the eyes twice a day for 28 days and 56 days. The placebo cream was applied contralaterally.

For the different time measurements of the study (T0, T28d and T56d), a photo workstation was used to obtain standardized photos. Then, the color of the black eye was analyzed.

The camera system used to take the standardized photographs included the flash system (R1C1Nikon TM) of a professional camera (D300Nikon TM) equipped with a macro lens (AF-S Micro NikkorTM60 mm).

In measuring dark circles, it is believed that the accumulation of hemoglobin in the eye contour region produces a color change. This change is related to the change in the red and blue components of the skin color (purple is a characteristic of the ring).

Then, image analysis software cielab (tm) that can be used in the reference space is used. The CIELab chromaticity space defines color by 3 coordinates:

l: from 0 (black) to 100 (white)

a is as follows: from 100 (red) to-100 (green)

b: from 100 (yellow) to-100 (blue).

In the present case, a decrease of + a and-b is expected.

The evaluation of the eye color is done by defining the area of the circle and comparing it to the nearby area (template in our case). In this way, Δ a for the red component and Δ -b for the blue component can be determined.

Results

TABLE 23：

After applying the cream of the invention for 30 days, the red component is reduced by 4.5 percent; compared to this, the side treated with placebo was increased by + 0.8%. The difference in action between the cream of the invention and placebo was evident, p < 0.05.

After 60 days, the effect of the cream of the invention is improved, with a significant reduction (p <0.05) of up to 7.5%; while the side treated with the placebo cream showed 3% degradation. The difference in action between the cream of the invention and the placebo cream is very clear, p < 0.01.

Watch 24：

30 days after application of the cream according to the invention, a reduction of the blue component of 3.2% was found; in contrast, the side treated with placebo was increased by 1.9%.

After 60 days, the effect of the product enables the blue component to be obviously reduced by 8.2 percent (p is less than 0.05); while the side treated with the placebo cream showed 4.1% degradation. The difference in action between the cream of the invention and placebo was evident, p < 0.05.

3.Testing of the Effect on the appearance of the eyes

The principle is as follows:the efficacy study of the cream of the invention was to demonstrate the perceived improvement in the appearance of their eyes (global fatigue, dark circles, bags, fine lines) in volunteers. For two-daily applications lasting 28 days, evaluations were performed every 2-3 days.

The study group was 105 white women, with a mean age of 51 years (between 40 and 60 years), alleged to have a fatigued appearance, and exhibited one of the following 2 to 3 characteristics: fine lines, bags, dark circles around the eyes. Volunteers applied the cream according to the invention on the contour of the eyes twice a day for 28 days.

The assessment is performed by self-assessment of the questionnaire in the morning, prior to the first application during the day.

Volunteers were asked to answer 4 questions:

1. how do you evaluate your eye fatigue when getting up? (Global)

2. How do you rate the importance of the presence of thin lines?

3. How do you evaluate the importance of the pouch?

4. How do you evaluate the importance of dark circles?

The answer (score) is: none (0), slight (1), moderate (2), important (3).

Question 1 was presented every monday, wednesday and friday (13 assessments over 28 days); questions 2, 3 and 4 were asked only once a week (5 assessments over 28 days).

Table 25:average assessment of eye appearance characteristics by the panelists.

The results are presented as average scores. At days 7, 14, 21, 28, the decrease in the mean value of the entries is expressed as% change. The statistical test is a t student test.

From an analysis of the above table of results, it can be said that:

the cream of the invention improves the appearance of the eyes when getting up, fatigue and dark circles, bags and fine lines. At the end of 28 days of use, the participants' assessment results were an average 42% to 46% reduction for the 4 listed entries. This reduction is statistically significant, with p < 0.01.

Fatigue of the eye appearance shows a clear reduction from day 2 of application (-9%, p <0.01, 2.7 phase vs 2.28) compared to the eye appearance at D0; at D7, it reached-16%.

The appearance of the dark circles, bags and strings showed a clear reduction after 7 days of application compared to that of D0 (16% for strings and bags, p < 0.01; 22% for dark circles, p < 0.01).

Claims (13)

1. Use of an extract of albizzia julibrissin dura as a topical cosmetic treatment for improving the phenomenon of skin fatigue selected from improving under-eye puffiness, under-eye dark circles and skin radiance, wherein the alcohol is methanol, ethanol, propanol, butanol or mixtures thereof, of an extract of flowers and/or seeds of albizzia julibrissin dura and/or alcohol.

2. Use according to claim 1, characterized in that the extract is diluted in a physiologically acceptable excipient.

3. Use according to claim 1, characterized in that the extract is diluted in a water-glycerol matrix.

4. Use according to any one of claims 1 to 3, characterized in that the extract combines one or more additional active ingredients selected among the following ranges: lightening, anti-redness, anti-pigmentation, calming, for treating allergic or reactive skin, UV sunscreens, moisturizing, moisturizers, exfoliates, smoothening, toning, anti-aging, detoxifying, anti-hair regrowth, acting on the skin barrier, anti-acne, acting on sebum secretion, matting, anti-inflammatory, anti-oxidant, anti-glycation, promoting blood circulation, peptides, vitamins active ingredients.

5. Use according to claim 4, characterized in that the additional active ingredient is siegesbeckia orientalis glycoside.

6. Use according to claim 5, wherein the darutoside is extracted from siegesbeckia orientalis.

7. Topical cosmetic treatment composition for improving the phenomenon of skin fatigue, comprising an extract of albizzia julibrissin and siegesbeckia orientalis glycosides in a physiologically acceptable medium, said extract being an extract of the flowers and/or seeds of albizzia julibrissin with water and/or an alcohol, wherein said alcohol is methanol, ethanol, propanol, butanol or a mixture thereof; the albizzia julibrissin extract or siegesbeckia orientalis glycoside is present in an amount ranging from 0.001% to 10% based on the total weight of the composition.

8. The composition of claim 7, wherein the darutoside is extracted from siegesbeckia orientalis.

9. Composition according to claim 7, characterized by comprising one or more additional active ingredients selected among the following ranges: lightening, anti-redness, anti-pigmentation, calming, for treating allergic or reactive skin, UV sunscreens, moisturizing, moisturizers, exfoliates, smoothening, toning, anti-aging, detoxifying, anti-hair regrowth, acting on the skin barrier, anti-acne, acting on sebum secretion, matting, anti-inflammatory, anti-oxidant, anti-glycation, promoting blood circulation, peptides, vitamins active ingredients.

10. A composition according to any one of claims 7 to 9 for the cosmetic treatment of skin fatigue.

11. A composition according to claim 10 for the cosmetic treatment of the contour of the eye.

12. A composition according to claim 11 for the cosmetic treatment of dark circles and bags under the eyes.

13. Use of an extract of albizzia julibrissin in association with siegesbeckia orientalis glycosides for the preparation of a topical cosmetic treatment composition for improving the phenomenon of skin fatigue, wherein the extract of albizzia julibrissin is an extract of the flowers and/or seeds of albizzia julibrissin with water and/or an alcohol, wherein the alcohol is methanol, ethanol, propanol, butanol or a mixture thereof.