FR3060568A1 - OLIGOPEPTIDE FRACTION OBTAINED FROM A BIOMASS OF BACTERIUM (S) BELONGING TO THE GENUS VITREOSCILLA SP. AS A COSMETIC ACTIVE - Google Patents

Abstract

The invention relates to an oligopeptide fraction obtained from a biomass of bacterium (s) belonging to the genus Vitreoscilla sp., As a cosmetic active agent, with particular cosmetic and / or dermatological compositions which contain it and its use. cosmetic and / or dermatological, preferably as a moisturizing, anti-aging and / or strengthening of the hair fiber.

Description

060 568

62668 ® FRENCH REPUBLIC

NATIONAL INSTITUTE OF INDUSTRIAL PROPERTY © Publication number:

(to be used only for reproduction orders)

©) National registration number

COURBEVOIE © Int Cl ⁸ : C07K4 / 00 (2017.01), A 61 K8 / 99, A 61 Q 19/08, C 12 N 9/24, 9/88

A1 PATENT APPLICATION

©) Date of filing: 16.12.16. (© Applicant (s): L'OREAL Société anonyme— FR. (30) Priority: @ Inventor (s): TOURNIER-COUTURIER LUCIE, YU PEIYAN and ROMAN BOY. (43 / Date of public availability of the request: 22.06.18 Bulletin 18/25. ©) List of documents cited in the report preliminary research: Refer to end of present booklet (© References to other national documents ©) Holder (s): L'OREAL Société anonyme. related: ©) Extension request (s): (© Agent (s): L'OREAL Société anonyme.

OLIGOPEPTIDE FRACTION OBTAINED FROM A BACTERIA BIOMASS (S) BELONGING TO THE GENUS VITREOSCILLA SP., AS A COSMETIC ACTIVE.

The invention relates to an oligopeptide fraction obtained from a biomass of bacterium (s) belonging to the genus Vitreoscilia sp., as a cosmetic active agent, to compositions in particular cosmetic and / or dermatological which contain it as well as to its cosmetic and / or dermatological use, preferably as a moisturizing, anti-aging and / or strengthening agent for the hair fiber.

FR 3 060 568 - A1

i

Oligopeptide fraction obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp., As a cosmetic active ingredient

Field of the invention

The present invention relates to active agents intended for use in the field of cosmetics and / or dermatology, preferably cosmetics.

More particularly, the invention relates to an oligopeptide fraction obtained from a biomass of bacterium (s) belonging to the genus Vitreoscilla sp., As a cosmetic active agent, to compositions in particular cosmetic and / or dermatological which contain it thus than for its cosmetic and / or dermatological use, preferably as a moisturizing, anti-aging and / or strengthening agent for the hair fiber.

Technological background

Over time, it appears that keratinous materials, such as the skin, the eyelashes, the eyebrows, the nails, the hair are subject to aging, resulting in particular in a modification of their structures and of their functions.

Endogenously, keratin materials are under attack by superoxide ions, naturally produced during physiological cellular metabolic processes.

Exogenously, keratin materials are exposed, on the front line, to environmental factors which participate in their aging, such as for example sunlight, in particular via ultraviolet radiation, or even pollution. These can disrupt the filaggrin cycle and therefore the production of natural hydration factors (NMFs). Consequently, the skin loses its natural humectants and becomes sensitive to desiccation. The barrier function is then weakened and altered, the skin is more exposed to external aggressions.

Extrinsic aging of the skin influenced by the environment, for example, adds to the intrinsic aging linked to age, giving rise to changes and characteristic skin damage, well described in the literature.

From a cellular and molecular point of view, aging of the skin results in an imbalance between the production of the constituents of the dermis, such as, for example, collagen and fibrillin, a decrease in markers of differentiation, such as, for example, transglutaminases ( TGM) and their destruction by the different metalloproteases (MMP) or genes responsible for the synthesis of hyaluronic acid such as the HAS 3 gene (Hyaluronan synthase), tipping the balance in favor of the degradation of the extracellular matrix and the destruction tissue.

From a macroscopic point of view, aging of the skin manifests itself in addition to the appearance of wrinkles and laxity. Aged skin is thinner and more fragile than young skin. Finally, aged skin tends to lose its elasticity, its barrier function against external aggressions, and its repair capacity.

Several compositions or methods are known in the state of the art for preventing and / or treating the signs of skin aging. Indeed, it is known from the state of the art the use of a total lysate of bacteria belonging to the genus Vitreoscilla sp., In a complete fermentation medium, as an active ingredient for preventing and / or treating hyperseborrhoeic states not associated with a dandruff condition of the scalp (FR 2 988 607) or also, as an active agent for preventing and / or treating dandruff conditions of the scalp in particular associated with a proliferation of pathogenic microorganisms on the scalp and / or an imbalance of the scalp ecoflora (FR 2 973 700).

The lysates described in these documents thus comprise a mixture of the protein, lipid and polysaccharide fractions in their culture medium and are not intended for hydration of the skin, for treatment and / or prevention of aging of the skin and / or strengthening the hair fiber.

Furthermore, the oligopeptide fraction according to the invention differs from the lipopolysaccharide extract described by document FR 2 838 056, in particular by its composition of oligopeptides.

For example, the use of an extract of non-photosynthetic non-fruiting filamentous bacteria has been proposed (FR 2 719 768 and FR 2 838 056). In this case, the prevention and / or treatment of the signs of skin aging is / are notably obtained with an extract of Vitreoscilla filiformis containing lipopolysaccharides, by inducing an increase in the endogenous synthesis of superoxide dismutase ( FR 2 838 056).

Furthermore, it has been described in document FR 2 762 782 that a clarified and stabilized culture medium of at least one non-photosynthetic filamentous bacterium has advantageous properties from the point of view of reduction and / or retardation skin aging.

In addition, in the hair sector, users are looking for products that improve the quality of hair. In particular, there is a growing need with regard to the lasting strengthening of the hair, in particular of fine or altered hair.

There is therefore a need to have new active agents or new treatments to prevent and / or repair the damage caused to keratin materials and more particularly to dehydrated, and / or aged skin, or at the level of the hair fiber.

There is a need for new cosmetic treatments capable of strengthening keratin materials and more particularly of anti-aging active agents capable of maintaining, or even strengthening the barrier properties of the skin, by repairing skin lesions as well as active agents capable of strengthening keratin fiber.

Surprisingly, the Applicant has demonstrated that the oligopetidic fraction according to the invention promotes hydration of the stratum comeum and therefore hydration of the skin. Likewise, the oligopeptide fraction makes it possible to increase the synthesis of pro-collagen 1, which demonstrates its activity in anti-aging. Furthermore, the oligopeptide fraction according to the invention increases the bending force of the hair fiber, reflecting a strengthening of the latter.

Summary of the invention

Consequently according to a first aspect, the invention relates to an oligopeptide fraction isolated from a lysate of a biomass of bacteria (s) belonging to the genus Vitreoscilla sp., According to another aspect, the invention relates to a process for the preparation of the oligopeptide fraction isolated from a lysate of a biomass of bacterium (s) belonging to the genus Vitreoscilla sp., comprising the steps consisting in:

(i) provide a biomass of bacteria belonging to the genus

Vitreoscilla sp. -, (ii) carry out a double enzymatic digestion of said biomass;

- (iii) inactivate enzymatic digestion;

- optionally clarify the reaction medium by one or more filtration steps; and

- optionally atomize the complete reaction medium, optionally clarified.

According to another aspect, the invention relates to an oligopeptide fraction isolated from a lysate of a biomass of bacterium (s) belonging to the genus Vitreoscilla sp., As cosmetic active, preferably as active for the reinforcement of keratinous matters and in particular as hydrating active, anti-aging and of reinforcement of the hair fiber.

Within the meaning of the invention, the term “keratin materials” means the skin and its appendages such as keratin fibers, in particular human keratin fibers such as the hair.

For the purposes of the present invention, the term "hair fiber" means the hair.

For the purposes of the present invention, the term “skin” means the skin of the body such as the skin of the hands, legs, arms, the skin of the face, the neck, and the scalp.

According to another aspect, the invention relates to a composition, in particular a cosmetic and / or dermatological composition comprising said oligopeptide fraction, according to the invention, obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp.

According to another aspect, the invention relates to a use of said composition, according to the invention obtained from a biomass of bacterium (s) belonging to the genus Vitreoscilla sp, as cosmetic active (and / or dermatological) ), preferably as an active ingredient for strengthening keratin materials and in particular as a hydrating, anti-aging active ingredient and for strengthening the hair fiber.

According to another aspect, the invention relates to a cosmetic process for treating keratinous material, comprising a step of topical application of the oligopeptide fraction isolated from a lysate of a biomass of bacteria belonging to the genus Vitreoscilla sp.

According to another aspect, the invention relates to a cosmetic method for moisturizing the skin, in particular dry skin, comprising a step of topical application of the oligopeptide fraction isolated from a lysate of a biomass of bacteria belonging to the genus Vitreoscilla sp.

According to another aspect, the invention relates to a cosmetic process for the treatment and / or prevention of the signs of aging of the skin, comprising a step of topical application of the oligopeptide fraction isolated from a lysate of a biomass of bacteria belonging to the genus Vitreoscilla sp.

According to another aspect, the invention relates to a process for strengthening the keratin fiber, comprising a step of applying the oligopeptide fraction isolated from a lysate of a biomass of bacteria belonging to the genus Vitreoscilla sp.

Detailed description of the invention

Oligopeptide fraction isolated from a lysate of a biomass of bacteria belonging to the genus Vitreoscilla sp.

By "oligopeptide fraction" is meant a fraction consisting of:

i) from 30 to 50% of oligopeptides, ii) between 1% and 10% of amino acids, iii) 10 to 20% of a mixture of oligosaccharides and monosaccharides.

Preferably the oligopeptides of the active have a molecular weight of between 200 and 3 kDa, with a peptide distribution centered on the zone 200-800Da; in particular, oligopeptides are peptides between 2 and 7 amino acids on average.

Glutamic acid, glutamine, histidine and arginine make up at least 15% of these 1-10% amino acids.

Preferably, the oligosaccharides are chosen from stachyose, sucrose, glucose, fructose and thehalose.

• Bacteria belonging to the genus Vitreoscilla sp. (including species:

Vitreoscilla filiformis)

As specified above, the microorganism considered according to the invention is a non-synthetic filamentous bacterium as defined according to the classification of Bergey's Manual of Systematic Bacteriology (Vol. 3, sections 22 and 23, 9 ^th edition, 1989), and belonging to the genus Vitreoscilla sp. (in particular species: Vitreoscilla filiformis).

More particularly, it is a bacterium belonging to the genus Vitreoscilla.

According to a preferred variant of the invention, it is the bacterium Vitreoscilla filiformis.

In a particular embodiment, the bacterium is the Vitreoscilla filiformis strain, preferably the Vitreoscilla filiformis ATCC 15551 strain.

• Complete fermentation medium

Within the meaning of the present invention, the expression “complete fermentation medium” is intended to denote a medium resulting from the culture process having been used for the growth and cell lysis of the microorganism, said medium having not otherwise undergone any additional manipulation aimed to separate and / or eliminate all or part of its non-aqueous constituents.

• Composition of fermentation medium

By definition, a fermentation or culture medium is a support which allows culture and therefore, as the case may be, the development of cells, bacteria, yeasts. In principle, the cells find in this medium the essential components for their multiplication in large numbers quickly, but also sometimes elements which will allow to favor the development of a specific bacterial genus or a particular family, in this case a bacterium. belonging to the genus Vitreoscilla sp. (in particular, species: Vitreoscillafiliformis).

Its composition must therefore meet the nutritional requirements of the microorganism considered and necessary for the proliferation thereof.

More specifically, the composition of this culture medium must:

- cover the needs in mineral ions, growth factors, provide the source of carbon and energy;

- present a pH close to the optimal pH

- present an optimal ionic strength (the medium can be isotonic but it is not compulsory).

Thus, the composition of a fermentation medium suitable for the invention comprises at least:

- a source of carbon and energy, generally represented by a sugar and advantageously glucose;

- a source of potassium and phosphorus such as K2HPO4, for example;

- A source of nitrogen and sulfur which can be represented by the compound (NH ₄ ) ₂ SO ₄ ;

- a source of magnesium such as for example MgCl ₂ ;

- a source of calcium such as for example CaCl ₂ ;

a source of iron and more particularly of iron citrate, the role of citrate being to maintain the iron in solution;

- a source of trace elements chosen in particular from Cu, Zn, Co salts,

Ni, B, Ti;

- a source of water, generally sterile, essential for all forms of life; and

- a pH buffer which can be represented by the KH ₂ PO ₄ .

In the absence of one of these components, bacteria do not grow, because they cannot by themselves make up for its absence.

As an illustration of a fermentation medium suitable for the development of a microorganism in accordance with the invention, mention may in particular be made of the medium appearing in Example 1 below.

An effective amount of the microorganism considered according to the invention is introduced into it and the whole is placed in conditions conducive to its proliferation.

• Digestion of bacteria to obtain an active ingredient according to the invention

Consequently, the oligopeptide fraction considered according to the invention can be obtained via a process consisting of:

- the culture of at least one bacterium belonging to the genus Vitreoscilla sp. (in particular species: Vitreoscilla filiformis) on a fermentation medium under conditions conducive to the proliferation of said bacteria, and

- a centrifugation step, and

- possibly pH adjustment, and

- the enzymatic digestion of the pellet resulting from the centrifugation of said bacteria in said fermentation medium.

The oligopeptide fraction within the meaning of the invention designates the active ingredient resulting from the digestion of the pellet resulting from centrifugation which is optionally adjusted to a given pH and subsequently treated with a carbohydrase enzyme and a protease enzyme.

According to a first embodiment, the digestion of the pellet resulting from centrifugation is, after possible adjustment of the pH to a given value, carried out sequentially by the action of a carbohydrase enzyme then by the action of a protease enzyme.

According to a second embodiment, the digestion of the pellet resulting from centrifugation is, after possible adjustment of the pH to a given value, carried out sequentially by the action of a protease enzyme then by the action of a carbohydrase enzyme.

According to another embodiment, an inactivation step, in particular thermal inactivation, takes place between the two steps of enzymatic digestion. According to a first variant of this embodiment, the pellet resulting from centrifugation is optionally adjusted to a given pH then first treated with a protease enzyme, then is subjected to an inactivation step, then is treated with a carbohydrase enzyme . According to a second variant of this embodiment, the pellet resulting from centrifugation is optionally adjusted to a given pH, then first treated with a carbohydrase enzyme, then is subjected to an inactivation step, then is treated with an enzyme protease.

The pH adjustment which can precede the enzymatic digestion step of the pellet is optional. When carried out, the pH is modified by adding an aqueous solution of acid or base, organic or mineral, the objective of this adjustment being to adjust if necessary the pH of the pellet to a value corresponding to the pH optimal use of the enzymes used in the digestion stage.

Preferably, the pH is adjusted. Preferably, the adjustment is carried out by adding an organic or mineral acid solution, preferably up to a pH of 3.5 to 5, more preferably from 3.8 to 4.2, in particular at a pH of 4.

By organic or mineral acid is meant, for example, carboxylic acids or sulfonic acids such as sulfonic acid, methanesulfonic acid, para-toluene sulfonic acid, camphorsulfonic acid, tartaric acid, citric acid, acetic acid, acid aspartic, glutamic acid, or mineral acids such as hydrochloric acid, sulfuric acid, phosphoric acid, phosphonic acid.

By enzyme carbohydrase is meant for example the enzymes of classification EC3.2.1. According to a preferred form of the invention, by enzyme carbohydrase means the enzyme Viscozyme® sold by the company Novozyme.

By protease enzyme is meant for example the enzymes of classification EC3.4.

According to a preferred form of the invention, the term “protease enzyme” means the enzyme Sumizyme AP sold by the company Novozyme.

According to a preferred embodiment, the digestion of the pellet resulting from centrifugation is carried out sequentially by the action of Viscozyme® marketed by the company Novozyme then by the action of Sumizyme AP marketed by the company Novozyme.

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To obtain an oligopeptide fraction obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp. in a complete fermentation medium, according to a process comprising a step of culturing said bacteria, the skilled person can refer in particular to Example 1.

The invention also relates to a process for the preparation of an oligopeptic fraction obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp., Comprising the steps consisting in:

(i) provide a biomass of bacteria belonging to the genus

Vitreoscilla sp.

(ii) carrying out a double enzymatic digestion of said biomass;

(iii) inactivate enzymatic digestion;

Preferably, step (iii) is followed by a step of atomizing the complete reaction medium.

The invention also relates to a process for the preparation of an oligopeptic fraction obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp., Comprising the steps consisting in:

- provide a biomass of bacteria belonging to the genus

Vitreoscilla sp. then

- possibly perform centrifugation

- possibly adjust the pH

- carrying out a double enzymatic digestion of said biomass;

- inactivate enzymatic digestion;

- possibly adjust the pH

- possibly clarify by successive filtration

- possibly atomize the complete reaction medium.

By "biomass" is meant all or part of the mass of living or dead organisms contained in a culture carried to completion, that is to say after disappearance of all or part of the nutrients constituting the initial complete fermentation medium.

In a preferred embodiment, the biomass is subjected to a centrifugation step prior to the double digestion step and the double digestion step is carried out on the pellet from the centrifugation.

The adjustment of the pH which can precede the step of double enzymatic digestion is optional. When carried out, the pH is modified by adding an aqueous solution of acid or base, organic or mineral, the objective of this adjustment being to adjust if necessary the pH of the pellet to a value corresponding to the pH optimal use of the enzymes used in the digestion stage.

Preferably, the pH is adjusted. Preferably, the adjustment is carried out by adding an organic or mineral acid solution, preferably up to a pH of 3.5 to 5, more preferably from 3.8 to 4.2, in particular at a pH of 4.

Preferably the pH is adjusted with an orgnaic acid such as citric acid.

In a particular embodiment, the double enzymatic digestion of the biomass of bacterium (s) belonging to the genus Vitreoscilla sp. is carried out using the enzymes carbohydrase and protease.

According to a first variant, the digestion is, after possible adjustment of the pH to a given value, carried out sequentially by the action of a carbohydrase enzyme then by the action of a protease enzyme.

According to a second variant, the digestion is, after possible adjustment of the pH to a given value, carried out sequentially by the action of a protease enzyme then by the action of a carbohydrase enzyme.

According to one embodiment, an inactivation step, in particular thermal inactivation, takes place between the two steps of enzymatic digestion. According to a first variant of this embodiment, the biomass which may be centrifuged, is optionally adjusted to a given pH and then first treated with a protease enzyme, then is subjected to an inactivation step, then is treated with a carbohydrase enzyme.

According to a second variant of this embodiment, the optionally centrifuged biomass is optionally adjusted to a given pH, then first treated with a carbohydrase enzyme, then is subjected to an inactivation step, then is treated with a protease enzyme.

According to a preferred form of the invention, by enzyme carbohydrase means the enzyme Viscozyme® sold by the company Novozyme. Viscozyme is a broad-spectrum endoprotease, purified and isolated from Bacillus liceniformis. The amount of carbohydrase used is 0.05 to 1.5% by weight, more preferably 0.2%. The working temperature is 50 ° C to 60 ° C, preferably 55 ° C; the implementation pH is between 4 and 5 (limits included) and the treatment time is 2 hours to 24 hours, preferably 3 hours.

By protease enzyme is meant for example the enzymes of classification EC3.4.

According to a preferred form of the invention, the term “protease enzyme” means the enzyme Sumizyme AP sold by the company Novozyme. Sumizyme is a broad spectrum endoprotease, purified and isolated from Aspergillu niger. The amount of protease used is 0.01 to 1% by weight, more preferably 0.04%. The processing temperature is from 50 ° C to 60 ° C, preferably 55 ° C; the implementation pH is between 3 to 5 (limits included), preferably from 3 to 4.5, and the treatment time is from 2 hours to 24 hours, preferably 3 hours.

According to a preferred embodiment, the digestion of the biomass is carried out sequentially by the action of Viscozyme® marketed by the company Novozyme then by the action of Sumizyme such as Sumizyne AP marketed by the company Novozyme.

According to a particular embodiment, the inactivation of the enzymatic digestion is a thermal inactivation which can be carried out for a period of between 10 minutes to 30 minutes, preferably a duration of 15 minutes.

In a particular embodiment, the inactivation of the enzymatic digestion is carried out at a constant temperature between 80 ° C and 90 ° C, preferably a temperature of 90 ° C.

By about 15 min, we mean a duration of 15 min ± 5 min.

In a particular embodiment, at the end of the inactivation step, the pH of the reaction medium is adjusted to 7 with an organic or inorganic base such as sodium hydroxide (NaOH).

In a particular embodiment, the reaction medium after double digestion is clarified by filtration. Preferably, this clarification step consists of double filtration, preferably by implementing filtration with a 10 pm bag filter and then with a K2 filter.

According to a particular embodiment, the atomization technique is used on the complete reaction medium which may be clarified and preferably on the clarified medium. In a particular embodiment, the atomization technique is carried out with a BUCHI mini spray dryer B-290 atomizer with the following parameters Pump 65%; INLET temperature 110 ° c to 230 ° C and preferably 210 ° C; OUTLET temperature from 80 ° C to 90 ° C and in particular 87 ° C; Aspirator 95; Q flow 45.

The conditions of the step of inactivating the enzymatic digestion can be adapted by a person skilled in the art.

According to a particular embodiment, the method for preparing an oligopeptic fraction obtained from a biomass of bacteria (s) belonging to the genus Vitreoscilla sp., Comprises the steps consisting in:

- provide a biomass of bacteria belonging to the genus

Vitreoscilla sp. \ then

- possibly perform centrifugation

- possibly adjust the pH

- carrying out a double enzymatic digestion of said biomass;

- inactivate enzymatic digestion

- adjust the pH to a physiological pH, that is to say a pH compatible with the application on keratin materials.

According to this embodiment, the oligopeptide fraction according to the invention can be packaged in sterile packaging, and / or be added with one or more preservatives such as phenoxyethanol.

According to another particular embodiment, the process for preparing an oligopeptic fraction obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp., Comprises the steps consisting in:

- provide a biomass of bacteria belonging to the genus

Vitreoscilla sp. - then

- possibly perform centrifugation

- possibly adjust the pH

- carrying out a double enzymatic digestion of said biomass;

- inactivate enzymatic digestion;

- possibly clarify by successive filtration

- atomize the complete reaction medium.

According to a preferred embodiment, the process for preparing an oligopeptic fraction obtained from a biomass of bacteria (s) belonging to the genus Vitreoscilla sp., Comprises the steps consisting in:

- provide a biomass of bacteria belonging to the genus

Vitreoscilla sp. - then

- perform a centrifugation, then

- carry out a pH adjustment by adding an organic or mineral acid such as hydrochloric acid or citric acid, preferably citric acid, to bring the pH to an acidic pH, preferably between 3.8 and 5, then

- carrying out a double enzymatic digestion of said biomass, preferably by sequentially implementing first a carbohydrase then a protease and more particularly by first implementing the Viscozyme® marketed by Novozyme then the Sumizyme AP marketed by Novozyme, in the conditions mentioned above; then

- inactivate enzymatic digestion, preferably thermally;

then

- clarify by successive filtration; then

- atomize the reaction medium.

Use of the oligopeptide fraction

The inventors have surprisingly observed that the oligopeptide fraction obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp. has interesting properties from the point of view of the care of keratin materials and in particular of the skin and / or the hair fiber.

As is apparent from the examples presented below, the inventors have unexpectedly discovered that the oligopeptide fraction obtained from a biomass of such a bacterium in a complete fermentation medium, exhibits unexpected cosmetic properties.

The invention relates to a use of the oligopeptide fraction from a lysate of a biomass of bacterium (s) belonging to the genus Vitreoscilla sp., According to the invention, as cosmetic and / or dermatological active agent, preferably as a moisturizing active, anti-aging active and / or strengthening of the hair fiber.

In the context of the invention, the term "oligopeptide fraction obtained from a biomass of bacteria (s) belonging to the genus Vitreoscilla sp." , the result of enzymatic digestion, of the pellet resulting from the centrifugation of the fermentation medium having served for the culture of the microorganism belonging to the genus Vitreoscilla sp. , by a carbohydrase enzyme and by a protease enzyme such as the carbohydrase enzyme

Viscozyme sold by the company Novozyme and the protease enzyme Sumizyme AP sold by the company Novozyme.

The term “cosmetic and / or dermatological active agent” means an active agent having beneficial properties after application to keratin materials, in particular the skin and / or the hair. In particular, it is a topical application.

• Composition

The invention also relates to a cosmetic and / or dermatological composition comprising the oligopeptide fraction obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp.

Preferably, the composition according to the invention also comprises a physiologically acceptable medium.

A "physiologically acceptable medium" is, according to the invention, a medium compatible with keratin materials of human beings, in particular the skin of human beings.

The physiologically acceptable medium is adapted to the nature of the support on which the composition is to be applied as well as to the form in which the composition is intended to be packaged, in particular solid or fluid at room temperature and atmospheric pressure.

The physiologically acceptable medium can in particular be a cosmetically acceptable medium.

By “cosmetically acceptable medium” is meant an odorless, colorless, or unpleasant appearance medium which does not generate discomfort for the user.

A composition containing the active ingredient according to the invention can be administered by or topically.

By "administer" is meant the fact of ingesting or applying externally (topically), a composition according to the invention for aesthetic purposes, that is to say in order to improve the appearance, texture of keratin materials and more particularly of the skin, a mucous membrane, the scalp of the hair and keratin fibers. In other words, within the meaning of the invention, the term "administer" does not include treatment for medical purposes.

Preferably, the composition according to the invention is a composition intended to be applied topically to the skin, or a hair composition.

A composition according to the invention advantageously comprises an amount of oligopeptide fraction obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp.

In certain embodiments, a composition according to the invention advantageously comprises an amount of oligopeptide fraction according to the invention ranging from 0.01 to 10% by weight, relative to the total weight of said composition, more preferably 0.05% to 10% by weight and more particularly from 0.06 to 5% relative to the total weight of the composition.

In other embodiments, a composition according to the invention advantageously comprises an amount of oilgopeptide fraction ranging from 1% to 5% by weight, relative to the total weight of said composition.

A composition according to the invention can therefore be presented in all the galenical forms normally available for the specific mode of administration selected.

The support can be of various nature depending on the type of composition considered.

As regards more particularly the compositions intended for administration by external topical route, they may be aqueous, hydroalcoholic or oily solutions, solutions or dispersions of the lotion or serum type, emulsions of liquid or semi-liquid consistency. of the milk type, obtained by dispersion of a fatty phase in an aqueous phase (O / W) or vice versa (W / O), or of suspensions or emulsions, of soft consistency, semi-solid or solid, of the cream type, aqueous or anhydrous gel, of microemulsions, microcapsules, microparticles, or vesicular dispersions of ionic and / or nonionic type.

These compositions are prepared according to the usual methods.

These compositions can in particular constitute creams for cleaning, protecting, treating or caring for lotions, gels or foams for caring for keratin materials such as the skin, the mucous membranes, the scalp.

They can be used for the cosmetic treatment of keratin materials, in particular of the skin, scalp and / or keratin fibers, in the form of solutions, creams, gels, emulsions, foams or also in the form of compositions suitable for the implementation of an aerosol or a spray, for example also containing a propellant under pressure.

In certain embodiments, a topical composition according to the invention can advantageously be formulated in any galenical form suitable for hair care, in particular in the form of a hair lotion, a shampoo, a conditioner, a detangler, a hair cream or gel, a styling hairspray, a styling lotion, a treating lotion, a dye composition (in particular of oxidation) optionally in the form of coloring shampoo, a restructuring hair lotion, a perm composition, a fall prevention lotion or gel, an antiparasitic shampoo, or a shampoo treating, in particular antiseborrheic, an anti-dandruff product , a scalp care product including anti-irritant, anti-aging, restructuring.

When a composition of the invention is an emulsion, the proportion of the fatty phase can range from 5 to 80% by weight, and preferably from 10 to 50% by weight relative to the total weight of the composition. The oils, emulsifiers and coemulsifiers used in the composition in emulsion form are chosen from those conventionally used in the cosmetic and / or dermatological field. The emulsifier and the coemulsifier may be present in the composition in a proportion ranging from 0.3 to 30% by weight, and preferably from 0.5 to 20% by weight relative to the total weight of the composition.

When the composition of the invention is an oily solution or gel, the fatty phase can represent more than 90% of the total weight of the composition.

In known manner, the dosage forms dedicated to topical administration can also contain conventional adjuvants in the cosmetic field, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents other than the oligopeptide fraction according to the invention, preservatives, antioxidants, solvents, fragrances, fillers, filters, odor absorbers and colorants.

The amounts of these various adjuvants are those conventionally used in the field under consideration, and for example from 0.01 to 20% of the total weight of the composition. These adjuvants, depending on their nature, can be introduced into the fatty phase and / or into the aqueous phase.

As emulsifiers which can be used in the invention, there may be mentioned for example glycerol stearate, polysorbate 60, cetylstearyl alcohol / cetylstearyl alcohol mixture oxyethylenated with 33 moles of ethylene oxide sold under the name Sinnowax AO® by the company HENKEL , the mixture of PEG-6 / PEG-32 / Glycol Stearate sold under the name Tefose® 63 by GATTEFOSSE, PPG-3 myristyl ether, silicone emulsifiers such as cetyldimethicone copolyol and sorbitan mono- or tristearate, PEG-40 stearate, oxyethylenated sorbitan monostearate (20OE).

As solvents which can be used in the invention, mention may be made of lower alcohols, in particular ethanol and isopropanol, propylene glycol.

The composition of the invention may also advantageously contain thermal and / or mineral water, in particular chosen from Vittel water, the waters of the Vichy basin and the water of La Roche Posay.

As hydrophilic gelling agents, mention may be made of carboxylic polymers such as carbomer, acrylic copolymers such as acrylate / alkylacrylate copolymers, polyacrylamides and in particular the mixture of polyacrylamide, C13-14-Isoparaffin and Laureth-7 sold under the name of Sepigel 305® by the company SEPPIC, polysaccharides such as cellulose derivatives such as hydroxyalkylcelluloses and in particular hydroxypropylcellulose and hydroxyethylcellulose, natural gums such as guar, locust bean and xanthan and clays.

As lipophilic gelling agents, mention may be made of modified clays such as bentones, metal salts of fatty acids such as aluminum stearates and hydrophobic silica, or alternatively ethylcellulose and polyethylene.

• Processes

The invention also relates to a cosmetic process for treating keratinous material, comprising a step of topical application of an oligopeptic fraction obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp.

In the context of the invention, by "treatment of a keratinous material" means the topical application of an oligopeptide fraction according to the invention for the care of the skin, mucous membranes, lips, nails, scalp and hair.

The invention also relates to a cosmetic process for hydration of the skin, comprising a step of topical application of an oligopeptide fraction obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp.

The invention also relates to a cosmetic process for the treatment and / or prevention of the signs of aging of the skin, comprising a step of topical application of an oligopeptide fraction obtained from a biomass of bacteria belonging to genus Vitreoscilla sp.

In the context of the present invention, by "treatment of the signs of aging of the skin" is meant the reduction or attenuation of wrinkles, fine lines, wilting of the skin, loss of elasticity of the skin, loss of skin tone, thinning of the dermis, breakdown of collagen fibers, loss of skin firmness.

In the context of the present invention, by “prevention of the signs of aging of the skin” is meant the absence of appearance or the delay in the appearance of wrinkles, fine lines, wilting of the skin, loss elasticity of the skin, loss of skin tone, thinning of the dermis, degradation of collagen fibers, loss of skin firmness ,.

The invention also relates to a cosmetic process for the treatment of the hair fiber and in particular for the strengthening of the hair fiber, comprising a step of applying to the hair fiber an oligopeptide fraction obtained from a biomass of bacteria ( s) belonging to the genus Vitreoscilla sp.

Figure 1: Induction of procollagen I

Examples

1- Preparation of an oligopeptide fraction within the meaning of the invention

Table 1: composition of the complete nutritive medium for the culture of V. fdiformis.

Chemical name [concentration] Autolytic yeast extract 4 g / 1 Papain soybean peptone F 3 g / 1 Glucose - Roférose 3 g / 1 KH ₂ PO ₄ 0.088 g / 1 CaCl ₂ 0.050 g / 1 CuSO ₄ , 5 H ₂ O 60 pg / l MnSO ₄ , 1H ₂ O 152 pg / l Kl 20 pg / l ZnSO ₄ , 7H ₂ O 200 pg / l A1C1 ₃ , 6H ₂ O 100 pg / l Osmotic water qs 1 1

To obtain a cosmetic active agent according to the invention, the procedure is as described below.

The initial strain of V. fdiformis is obtained from the ATCC (strain

15551).

The biomass is obtained by fermentation in an industrial bioreactor of

3000 liters effective, the composition of the culture medium is given above. During cultivation, the pH is kept constant (7.00), as well as the temperature (26 ° C) and the dissolved oxygen (0.5%). The culture is stopped when the dry matter content reaches 0.6% and the glucose content reaches 0.035%. Fermentation of strain V.

fdiformis is carried out in its complete culture medium.

The whole is centrifuged and the pellet is treated by enzymatic digestion.

Carbohydrase is first introduced for enzymatic digestion

Viscozyme® marketed by Novozyme in an amount of 0.2% w / w for a period of 3 hours at a temperature of 55 ° C the pH is adjusted and maintained at 4.5 with citric acid, then a Sumizyme AP protease marketed by Novozyme is introduced in an amount of 0.04% w / w for a period of 3 hours at a temperature of 55 ° C. the pH is maintained at 4.5 after adjustment by adding citric acid if necessary and.

Following this second enzymatic digestion, the medium is neutralized to pH 7 using a sodium hydroxide solution (NaOH). The activation of the enzyme is carried out by heating the medium to a temperature of 90 ° C for 15 minutes.

The neutralized and inactivated medium is clarified by successive filtrations first with a 10pm bag filter then a K2 filter.

A step of atomizing the complete clarified reaction medium is carried out with a BUCHI mini spray dryer B-290 atomizer with the following parameters Pump 30%; INLET temperature 160 ° C; OUTLET temperature 85 ° C; Aspirator 95; Q flow 35. The active ingredient is then recovered in the form of a powder and is used as it is.

2) preparation of a V. filiformis biomass extract

The Vitreoscilla filiformis strain (ATCC 15551) is cultured according to the method described in patent application WO-A-94-02158.

It is a continuous cultivation process. The culture is carried out at 26 ° C for at least 48 hours until a suitable cell concentration corresponding to an optical density at 600 nm greater than or equal to 1.5 is obtained. The strain is subcultured at 2% V / V in new medium for approximately 48 hours until a stable culture is obtained. A 1 liter Erlenmeyer flask containing 200 ml of new medium is then inoculated with 4 ml of the previous culture.

The Erlenmeyer culture is carried out at 26 ° C. on a culture table stirred at 100 rpm. The base stock thus obtained serves as an inoculum for a 50 liter fermenter. Growth takes place at 26 ° C, pH 7, 100 revolutions / minute and pO ₂ > 15%.

After 30 hours of growth, the biomass is transferred to a 3000 liter useful fermenter, to be cultivated under the same conditions. After 48 hours of growth, the cells are harvested continuously. The biomass is then concentrated approximately 50 times by centrifugation. The cells obtained are then frozen as the culture continues. These cells can be used as they are after thawing (non-stabilized cell extract) or else be stabilized by autoclaving at 121 ° C for 20 to 40 minutes (stabilized cell extract). The cells then burst during sterilization by releasing the cytosol and agglomerating the proteins as well as the walls. The product obtained is then biphasic.

The supernatant liquid phase can be filtered at 0.22 μm to remove the particles ("supernatant").

The bacteria extract, in the form of a cell extract (stabilized or not) or of a supernatant, can be used as it is (aqueous form) or can be lyophilized according to conventional techniques (lyophilized form).

3 / Effect of an oligopeptide fraction according to the invention ‘example 1) on the hydration of the Stratum Corneum and comparison with extract Vf outside the invention described in example 2.

A test was carried out to evaluate the hydration potential of the compounds of the invention which are formulated in an aqueous solution in an amount of 4% by weight relative to the total weight of the composition.

The technique allows measurement of the dielectric capacitance of the stratum corneum (SC) which depends on the average value of dielectric permittivity of the tissue. Dielectric permittivity varies greatly with the amount of water in the Stratum Corneum (SC).

The CS samples are conditioned at 75% relative humidity and at 25 ° C before / during the measurements and processing. The capacitance measurement is carried out using Coméométrie ™ (Courage & Khazaka, Germany).

The compound tested according to the invention and the comparative active ingredient (Vf) is dissolved in a water / n-propanol mixture (80/20) and the solution is deposited on the SC at the rate of 10 μL / cm ² before drying with l air lasting a total of 1.5 hours.

A measurement is carried out at T0, before the treatment, and a Tirait measurement is carried out after the treatment has completely dried.

Each treatment is systematically compared with its control (vehicle) and its TO. Using at least two different lots of SC. 4 to 5 SC samples are measured per treatment.

The change in the cometometer signal (HCM) after treatment is first calculated for each sample of SC: DHCMi = HCMi (Ttrait) -HCMi (TO). The average of the variations

DHCMi (vehicle) is then calculated for the control samples (treated with the vehicle): this average value is subtracted from all the variations DHCMi (active) to correct the systematic bias.

For each sample i, we measure:

- For the vehicle (witness): DHCMi (veh) = HCMi _ve h (Ttrait) - HCMi _ve h (T0)

- For the active: DHCMi _active = HCMi _active (Ttrait) - HCMi _active (T0)

To correct the systematic bias linked to the vehicle, the corrected value DHCMi _act if _CO rr according to: DHCMi _act if _CO rr = DHCMi _act if - M, where M corresponds to the average of the DHCMi _ve h variations observed on the n control samples:

The DHCMi _act if _C orr mean values and the associated standard deviations which have been obtained are shown in the table below.

Product concentration Change in value of comometer Average (n = 4) Standard deviation vehicle (milliQ / n water- propanol, 8: 2 flight) 2.9 6.7 fraction oligopeptide according to the invention (example 1 solubilized in water at 4% active ingredient) 4% MA 14.9 3.3 extract of Vitreoscilla filiformis out invention (example 2) 4% MA 3.1 2.8

It emerges from this test shows that the dielectric capacitance of the SC is much better with the oligopeptide fraction of the invention in comparison with that obtained with the Vitreoscilla filiformis extract of the prior art described in Example 2 at the same concentration in active ingredient.

The oligopeptide fraction according to the invention allows good hydration of the SC and therefore of the skin.

4 / Effect of an oligopeptide fraction according to the invention (example 1), on skin aging.

Stimulation of pro-collagen synthesis I

Biological model

Normal human fibroblasts are cultured and seeded in a 96 microtiter plate with 3500 cells per well. The cells are cultured for 6 hours in complemented MEM culture medium.

Treatment

The compound of the invention (Example 1) is added to the cells during the renewal of the culture medium at several concentrations. The cells are incubated for 5 days. After incubation, the culture medium is separated from the cells.

ELISA assay

- The culture medium is used for the quantification of procollagen I by ELISA.

The antibody used for the capture will be an anti ProCollagen Type I antibody from the supplier is US Biological (reference = P9005-74D)

- The antibody used for the detection will be an anti ProCollagen Type I Propeptide antibody from the supplier is US Biological (reference = P9005-74E)

Cell counting

The number of cells is counted by a spectrofluorimeter after a pretreatment of the cells with the reagent CyQuant.

Result

At 0.08g / L, the compound of the invention stimulates the production of pro-collagen by a factor of 2: EC2.0 = 0.08g / L (Figure 1)

Claims (18)

1. Oligopetidic fraction isolated from a lysate of a biomass of bacteria belonging to the genus Vitreoscilla sp. 2. The oligopeptide fraction according to claim 1, which comprises i) from 30 to 50% of oligopeptides, ii) between 1% and 10% of amino acids, iii) 10 to 20% of a mixture of oligosaccharides and monosaccharides 3. Oligopeptide fraction according to any one of the preceding claims wherein said bacterium belonging to the genus Vitreoscilla sp. is the V.filiformis strain, preferably the V.filiformis ATCC 15551 strain. 4. Oligopeptide fraction according to any one of the preceding claims, characterized in that it is obtained by a step of enzymatic digestion of said biomass. 5. Composition comprising, in a physiologically acceptable medium, an oligopeptide fraction according to any one of claims 1 to 4. 6. Use of an oligopeptide fraction obtained from a biomass of bacteria belonging to the genus Vitreoscilla sp., As a cosmetic active. 7. Use of an oligopeptide fraction according to claim 6 wherein said cosmetic active is a moisturizing active. 8. Use of an oligopeptide fraction according to claim 6 wherein said cosmetic active is an anti-aging active. 9. Use of an oligopeptide fraction according to claim 6 wherein said cosmetic active is an active for the strengthening of the hair fiber. 10. Use according to any one of claims 6 to 9, in which said oligopeptide fraction is obtained by a step of enzymatic digestion of said biomass. 11. Cosmetic method for treating a keratinous material, comprising a step of applying an oligopeptide fraction according to any one of claims 1 to 4, or a composition containing it according to claim 5. 12. Cosmetic method for hydration of the skin, comprising a step of topical application of an oligopeptide fraction according to any one of claims 1 to 4, or a composition containing it according to claim 5. 13. Cosmetic process for the treatment and / or prevention of the signs of aging of the skin, comprising a step of topical application of an oligopeptide fraction according to any one of claims 1 to 4, or a composition containing it according to the claim 5. 14. Cosmetic method for strengthening the hair fiber, comprising a step of topically applying an oligopeptide fraction according to any one of claims 1 to 4, or a composition containing it according to claim 5. 15. Process for the preparation of an olgopeptide fraction obtained from a lysate of a biomass of bacteria belonging to the genus Vitreoscilla sp., Comprising the steps consisting in: (i) provide a biomass of bacteria belonging to the genus Vitreoscilla sp. ; (ii) carrying out a double enzymatic digestion of said biomass; (iii) inactivate enzymatic digestion. 16. The preparation method according to claim 15 characterized in that step (iii) is followed by a step of atomizing the complete reaction medium. 17. The preparation method according to claim 16, wherein said enzymatic digestion is carried out using carbohydrase and protease enzymes. 18. A preparation method according to claims 16 and 17, wherein said enzymatic digestion is carried out from the enzymes carbohydrase in a first step then protease in a second step. Nltrton Beu 'Induction Piocoliaoene i