WO2002015871A1

WO2002015871A1 - Use of tyramine in cosmetic compositions to lighten the skin

Info

Publication number: WO2002015871A1
Application number: PCT/FR2001/002597
Authority: WO
Inventors: Karl Lintner
Original assignee: Sederma S.A.
Priority date: 2000-08-25
Filing date: 2001-08-10
Publication date: 2002-02-28
Also published as: FR2813188B1; JP2004506677A; JP4393064B2; FR2813188A1; AU2001284135A1

Abstract

Tyramine can be advantageously used in any cosmetic or dermopharmaceutical composition for skin care, particularly for lightening it and for lessening its colouring when exposed to natural or artificial UV. The tyramine used can be derived from plant extract or from a chemical synthesis process. It is used as such or purified, vectorized or not in all types of carriers, including liposomes. It can be used on its own or combined with other active molecules, grafted or not by any alkyl chain, linear or branched, saturated or unsaturated, hydroxylated or sulphured or not, containing 1 to 24 carbon atoms.

Description

Use of tyramine in cosmetic compositions intended to lighten the skin. Melanin, synthesized in specialized cells of the epidermis, is responsible for pigmentation and therefore, the color of the skin. In normal skin, this synthesis is regulated by various factors, the best known of which is the sun, which increases it in proportion to the amount of UV radiation received during exposure (normal life or natural sunbathing or exposure sessions in a beauty salon) and whose manifestation is tanning. More or less benign or physiological disturbances of melanogenesis manifest themselves as freckles, moles or as diffuse spots (chloasma, pregnancy spots).

Among many natural physiological disturbances induced by aging, that of melanogenesis results in the appearance of heterogeneous zones which form spots of senescence.

The Cosmetic Industry is therefore completely in its role when it offers solutions capable of regulating these various dysfunctions which have no vital prognosis. Many molecules or substances have been proposed to treat hyperpigmentation of the skin, but few of them can be retained for a topical cosmetic application (problems of irritation, legislation, toxicity).

Those that remain after this selection are generally not very effective. The object of this patent application lies in the fact that we have discovered that it was possible to respond effectively to this problem by using tyramine in cosmetic or dermo-pharmaceutical compositions. To date, tyramine, which is mainly found in food (fermented cheeses, cabbage, potatoes, herring, gruyère, chianti, Camembert ...), is only very little used in pharmacies or in Cosmetic. It is used in dosage and diagnostic methods (EP0795610), to treat depressive states (US4868218) and / or Alzeimer or Parkinson's diseases (EP0406488), or to reduce the synthesis of cholesterol (US4857522).

From a biochemical point of view, tyramine is a direct precursor of dopamine. We hypothesized that, due to its chemical structure close to dopa, tyramine could present an anti-tyrosinase effect, which would make it a product capable of decreasing melanogenesis in all the situations described above.

In fact, melanocytes have a tyrosinase and an o-diphenol oxidase, enzymes which catalyze the conversion of tyrosine to melanin according to the following scheme:

Tyrosine = Dopa => Dopa quinone => Dopachrome = Indole quinone => Melanin

An albino individual has a congenital deficit of o-diphenol oxidase, which results in the absence of melanogenesis and therefore of skin pigmentation.

The invention which is the subject of this application resides in the fact that we have discovered and demonstrated that tyramine reduces the production of melanin because it is not transformed into dopa, which blocks the system. Tyramine is therefore a competitive inhibitor without being an alternative substrate.

Tyramine can be obtained from any source.

It may be advantageous to use a plant origin from any plant containing it, including Citrus reticulata, a plant particularly rich in tyramine (Wheaton

TA & Stewart I (1970) Lloydia 33: 244-54) since the whole plant or some isolated part can be used to extract it.

The plant or part of plant (15 grams) is dispersed in 85 grams of distilled water under blades for 48 hours at room temperature. The quantity of plant occupies as much volume as water. After filtration (800 μm) the extract can be used as it is.

It is also possible, for the purposes of subsequent formulations, to obtain a dry extract by any conventional method in the matter.

The analyzes carried out by HPLC show that the extract thus obtained contains tyramine. Depending on the origin of the plants and the harvest season, the skilled person adjusts the proportions given above to obtain extracts whose tyramine content is constant from one extraction to another.

This example of obtaining tyramine in a Citrus reticulata extract is not limiting.

It is indeed possible to produce this extract by other processes such as, for example, maceration, simple decoction, leaching, extraction under reflux, extraction by means of ultrasound or microwaves or finally counter-current techniques, without this list being exhaustive.

Furthermore, the extraction solvents mentioned above are not limiting and can be chosen from water, propylene glycol, butylene glycol, glycerin, polyethylene glycol, methyl or ethyl ethers of diglycols, cyclic polyols, ethoxylated or propoxylated diglycols, alcohols (methanol, ethanol, propanol, butanol), or any mixture of these solvents.

In summary, in the context of this patent, to respond to the expected activities, tyramine can:

• come from plant extracts, preferably Citrus reticulata,

• come from any synthesis process,

• be used alone or in combination with other active molecules,

• be purified and vectorized in all supports including liposomes, • be grafted, on OH or on NH ₂ , by any alkyl chain, linear or branched, saturated or unsaturated, hydroxylated or sulfurized or not, containing 1 to 24 atoms of carbon. Tyramine can be present in all dosage forms used in Cosmetics and dermopharmacy. The following two compositions are given by way of example only. Example No. 1 - Day cream

Volpo S20 2.4

Volpo S2 2,6

Prostearyl 15 8.0

Beeswax 0.5

Abil ^® ZP2434 3.0

Propylene glycol 3.0

Carbopol ^® 941 0.25

0.25 triethanolamine

Tyramine 5.0

Water & preservatives QSP 100g

Example N ° 2 - Deodorant and lightening body

Crillet3 2.5

Novol 0.9

Fluilan 2.5 Carbopol 940 0.3

Beeswax 2.0

Triethanolamine 0.1 Glycerin 5.0

Tyramine 3.0

Water & preservatives QSP 100g

The following examples will demonstrate the lightening effect of tyramine. Example No. 3 - Inhibition of human tyrosinase (in vitro)

This example highlights dopamine oxidase activity.

Normal human melanocytes (MNH) in culture are lysed by an Nepal buffer associated with ultrasound and the cell extract is brought into contact with a buffer containing tyramine, at concentrations of 0.1, 0.3 and 1 % (w / w) or containing none (controls) The activity of tyrosinase is then quantified through its L-Dopa oxidase activity by measuring the OD at 490 nm of the amount of Dopachrome formed from from L-

Dopa. The reaction is therefore initiated by the addition of L-DOPA.

The kinetics are monitored at a wavelength of 490 nm every 10 minutes between

T0 and 1H then every 30 minutes up to 2 Hours. The following table summarizes the average results of the inhibition percentages, observed with respect to the control at the same time, obtained on 8 different tests.

+ 10 '+ 20' +30 '+40' +50 '+60' +90 '+120

Tyramine 0.1% -13.2 -13.2 -16.1 -14.1 -13.4 -14.2 -14.0 -13.2

Tyramine 0.3% -19.2 -21.4 -28.3 -31.1 -32.1 -33.7 -33.5 -31.7

Tyramine 1.0% -27.0 -35.8 -48.4 -50.9 -52.8 -55.3 -55.7 -53.8

These results demonstrate a significant decrease in the activity of tyrosinase in the presence of tyramine. As the presence or absence of this tyramine is the only variable in the experiment described, it is clear that the effect observed can only come from tyramine. Finally, this effect is specific to the system observed with tyramine since the effect observed is clearly concentration-dependent.

It is understood that we have validated this trial in addition by ensuring the stability of the system by carrying out cases without L-Dopa and without tyramine.

Example No. 4 - Inhibition of tyrosinase (in vitro)

While it is intellectually satisfactory to use human cells to demonstrate a physiological effect, it is well known that the results obtained are sometimes subject to great variations due to the inter-individual variability of the donors. For this reason, although the previous results leave no doubt about the inhibitory effect of tyramine on tyrosinase, we have carried out different experiments with another model.

The cells used in this example are those of melanomas B 16 which produce a lot of melanin and which are commonly used to study the activity of tyrosinase as well as that of the synthesis of melanin.

These cells are reseeded into wells of culture plates (24 wells / plate) and incubated for 24 hours at 37 ° C. After rinsing, the cells are brought into contact, for 48 hours, with a buffer containing tyramine, at concentrations of 0.01 and 0.03% (w / w) or not containing it (controls).

A positive control was made using 0.01.10 ^"2 % hydroquinone.

As before, the cells are then lysed by an Igpal buffer associated with ultrasound, the activity of tyrosinase is quantified and standardized with respect to the quantity of proteins present.

Averages of inhibition percentages, finally observed incubation (48 H) compared to control at the same time (5 different trials)

Tyramine 0.01% - 22.1 Tyramine 0.03% - 29.9

These results demonstrate a significant drop in the activity of tyrosinase in the presence of tyramine which is better than that obtained with hydroquinone at comparable concentration. As the presence or absence of this tyramine is the only variable in the experiment described, it is clear that the effect observed can only come from tyramine. Finally, although achieved with only 2 different concentrations of tyramine, this effect is specific to the system observed with tyramine since the effect observed is concentration-dependent.

Example No. 5 - Inhibition of melanin synthesis (in vitro) Normal human melanocytes (MNH) in culture (confluence between 60 and 80%) are placed in the presence of buffer of a buffer containing tyramine, at concentrations 0.003 and 0.01% or not containing it (controls).

This contact time lasts 8 days during which, to maintain the cells, the buffer (identical to the previous one in terms of tyramine concentration: 0.003 and 0.01% (w / w)) is renewed every day. During this period, half of the boxes are exposed to UV-B 5 times

(20mJ / cm ² ) while the other half does not receive irradiation (controls).

At the end of this period, the culture medium is removed and the cells are rinsed.

After extraction, the quantity of intracellular melamine is conventionally determined by spectrophotometry and this quantity is reduced to the same number of cells for standardization.

The following table summarizes the average results of the inhibition percentages, observed compared to the controls at the same time, obtained on 5 different tests.

Non-irradiated cells Irradiated cells

Tyramine 0.003% - 30 - 28

Tyramine 0.01% - 32 - 32

Arbutin 0.09% - 11 - 8

These results demonstrate:

On the one hand, that in the absence of irradiation by UVB, tyramine decreases the basal level, which translates into a reduction in the total amount of melanin formed, hence an expected whitening effect,

• on the other hand, that after the 5 UV-B irradiations, the inhibition of melamine synthesis in the presence of tyramine is kept lower than in the controls, thus limiting the inducing effect of the radiation on neosynthesis ( factor * 2 in the case of controls).

As the presence or absence of this tyramine is the only variable in the experiment described, it is clear that the effect observed can only come from tyramine. Finally, although achieved with only 2 different concentrations of tyramine, this effect is specific to the system observed with tyramine since the effect observed is concentration-dependent.

Example No. 6 - "Anti-stain" effect (in vivo)

This test was carried out on 20 female volunteers aged 45 to 65 with age spots on the skin of the hands.

After precise identification and measurement of the size and coloration of 5 spots by hands and by volunteers, the protocol consists in applying the product to be tested incorporated (5%) in the cream described in example No. 1 for a period of '' a month and a half on one hand (drawn at random), the other serving as a control (therefore treated with the same cream but not containing tyramine). Obviously, measurements of the skin color of 5 areas without spots were also carried out to allow a standardization of the results.

At times + 1 month and + 1 and a half months, quantifications of the skin color are carried out using a mexameter which estimates the amount of melanin present thanks to a special lamp integrated in the probe and which emits 3 wavelengths different.

After a month and a month and a half, the coloration of the age spots on the treated hands was 6.4% and 24% respectively lower than those treated with the cream that did not contain tyramine. This result is particularly spectacular when we know that this study was carried out during the spring 2000 period which was particularly sunny.

All these examples clearly demonstrate a lightening and whitening effect as well as a real effectiveness on a lesser skin coloration during exposure to natural or artificial UV rays. The concentration of tyramine can vary between 0.001% and 10% (w / w), preferably between 0.01 and 7.0% (w / w) in the finished cosmetic or dermopharmaceutical composition.

Tyramine can be used in any dosage form used in cosmetics or dermopharmacy: O / W and W / O emulsions, milks, lotions, ointments, hair lotions, shampoos, soaps, sticks and pencils, sprays, body oils, without this list is limiting.

It is possible to incorporate tyramine into cosmetic vectors such as liposomes, chylomicrons, macro-, micro- and nanoparticles as well as macro-, micro- and nanocapsules, to absorb them on powdery organic polymers, talcs, bentonites and other mineral supports. Tyramine can be combined in cosmetic compositions with any other ingredient usually used in cosmetics: extraction and / or synthetic lipids, gelling and viscosifying polymers, surfactants and emulsifiers, hydro- or liposoluble active principles, extracts from other plants, tissue extracts, marine extracts.

The cosmetic or dermopharmaceutical compositions containing tyramine are creams, balms or gels or lotions or sunscreens and after sunscreens, after-shave products, depilatory or post-depilatory creams Tyramine is incorporated into any cosmetic or dermopharmaceutical composition intended for skin care, particularly for its whitening and its less coloring when exposed to natural or artificial UV rays.

These cosmetic or dermopharmaceutical compositions are used for the preparation of medicaments intended for skin care, in particular for its whitening and its less coloring when exposed to natural or artificial UV rays.

Tyramine is used alone or incorporated in cosmetic or pharmaceutical compositions, bound or incorporated) or absorbed or adsorbed in the form of macro-, micro- and nanoparticles or in macro-, micro- and nanocapsules, in textiles, synthetic or natural fibers, wool and any material that can be used to make clothes and underwear for day or night, directly in contact with the skin or hair to allow a topical delivery keep on going.

Claims

1. Use of tyramine in any cosmetic or dermopharmaceutical composition.

2. Cosmetic or dermopharmaceutical compositions according to claim 1, characterized in that the tyramine used can: • come from plant extract, preferably Citrus reticulata,

• come from any synthesis or fermentation process,

• be used alone or in combination with other active molecules,

• be purified and vectorized in all media including liposomes,

• be grafted, on OH or on NH ₂ , by any alkyl chain, linear or branched, saturated or unsaturated, hydroxylated or sulfurized or not, containing 1 to 24 carbon atoms.

3. Cosmetic or dermopharmaceutical compositions according to claim 2, characterized in that the concentration of tyramine is between 0.001% and 10% (w / w), preferably between 0.01 and 7.0% (w / w) in the product final.

4. Cosmetic or dermopharmaceutical compositions according to claims 2 to 3, characterized in that they are in any galenic form used in cosmetics or dermopharmacy: HZE and EH emulsions, milks, ointments, hair lotions, shampoos, soaps, sticks and pencils, sprays, body oils.

5. Cosmetic or dermopharmaceutical compositions according to one of claims 2 to 4, characterized in that tyramine is incorporated in cosmetic vectors such as liposomes, chylomicrons, macro-, micro- and nanoparticles as well as macro-, micro - and nanocapsules, to absorb them on powdery organic polymers, talcs, bentonites and other mineral supports.

6. Cosmetic or dermopharmaceutical compositions according to one of claims 2 to 5, characterized in that tyramine is used with any other ingredient usually used in cosmetics: extraction and / or synthetic lipids, gelling and viscosifying polymers, surfactants and emulsifiers, water- or fat-soluble active ingredients, extracts from other plants, tissue extracts, marine extracts.

7. Cosmetic or dermopharmaceutical composition according to one of claims 2 to 6, characterized in that they are creams, balms or gels or lotions or sunscreens and after sunscreens, aftershave products, depilatory or post creams - depilatory.

8. Cosmetic or dermopharmaceutical composition according to one of claims 2 to 7, intended for skin care, particularly for its whitening and lightening as well as for obtaining a lesser skin coloration during exposure to natural or artificial UV rays.

9. Use of the only tyramine according to claim 1 or incorporated in cosmetic or dermopharmaceutical compositions according to one of claims 2 to 7, for the preparation of medicaments intended for skin care, particularly for its whitening and its less coloring during natural or artificial UV exposure.

10. Use of the only tyramine according to claim No. 1 or incorporated in cosmetic or dermopharmaceutical compositions according to claims 2 to 7, bound (es) or incorporated) or absorbed (s) or adsorbed (es) in the form of macro- , micro- and nanoparticles or in macro-, micro- and nanocapsules, in textiles, synthetic or natural fibers, wool and any material that can be used to make clothes and underwear for day or night, directly to the skin or hair contact to allow continuous topical delivery.