FR2813188A1 - USE OF TYRAMINE IN COSMETIC COMPOSITIONS FOR CLEARING THE SKIN - Google Patents

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

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Melanin, synthesized in specialized cells of the epidermis, is responsible for the 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 causes the increase in proportion to the amount of UV radiation received during the exposure (normal life or natural sunbaths or exhibition sessions in 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 is reflected by the appearance of heterogeneous zones that form the spots of senescence.

The Cosmetic Industry is therefore entirely in its role when it proposes solutions to regulate these various dysfunctions that are not prognostic.

Many molecules or substances have been proposed to treat hyperpigmentation of the skin, but few of them can be retained for cosmetic topical application (irritation problems, legislation, toxicity).

Those remaining after this selection are generally not very effective.

The object of this patent application lies in the fact that we have discovered that it is possible to respond effectively to this problem by using tyramine in cosmetic or dermopharmaceutical compositions.

To date, tyramine, which is found mainly in food (fermented cheeses, cabbages, potatoes, herrings, Gruyere, Chianti, Camembert ...), is used very little in Pharmacy or Cosmetic. It is used in assay and diagnostic methods (EP0795610), for treating depressive states (US4868218) and / or Alzeimer or Parkinson's diseases (EP0406488), or for reducing cholesterol synthesis (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 have an anti-tyrosinase effect, which would make it a product capable of reducing melanogenesis in all the situations described above.

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 Indeed, melanocytes have tyrosinase and o-diphenol oxidase, enzymes that catalyze the conversion of tyrosine to melanin according to the following scheme: Tyrosine => Dopa => Dopa quinone => Dopachrome = * Indole quinone => Melanin An individual albino has a congenital deficiency of o-diphenol oxidase, which results in the absence of melanogenesis and therefore skin pigmentation.

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

Tyramine can be obtained from any source of supply.

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 1 (1970) Lloydia 33: 244-54) since it is possible to use the whole plant or some isolated part to make the extraction.

The plant or plant part (15 grams) is dispersed in 85 grams of distilled water under blades for 48 hours at room temperature. The amount of plant occupies as much volume as water. After filtration (800pm) the extract is usable as is.

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

The analyzes carried out by HPLC show that the extract thus obtained contains tyramine. Depending on the origin of the plants and the season of harvest, those skilled in the art adjust 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 achieve this extract by other methods such as, for example, maceration, simple decoction, leaching, extraction under reflux, extraction by means of ultrasound or microwaves or finally countercurrent techniques, without this list being limiting.

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

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 In summary, in the context of this patent, to respond to the expected activities, tyramine can: # come from plant extract, preferably from Citrus reticulata, # come from any synthesis process, # be used alone or in combination with d other active molecules, # be purified and vectorized in all supports including liposomes, # be grafted onto OH or NH2, by any linear or branched, saturated or unsaturated, hydroxylated or sulfur-containing alkyl chain, containing 1 at 24 carbon atoms.

Tyrarnine can be present in all galenic forms used in cosmetics and dermopharmacy. The following two compositions are given by way of example only.

<tb> Example <SEP> N <SEP> 1- <SEP> Cream <SEP> of <SEP> iour
<tb> Volpo <SEP> S20 <SEP> 2,4
<tb> Volpo <SEP> S2 <SEP> 2,6
<tb> Prostearyl <SEP> 15 <SEP> 8.0
<tb> Bee <SEP> wax <SEP> 0,5
<tb> Abil%) <SEP> ZP2434 <SEP> 3.0
<tb> Propylene <SEP> Glycol <SEP> 3.0
<tb> Carbopol * <SEP> 941 <SEP> 0.25
<tb> Tnethanolamine <SEP> 0.25
<tb> Tyramine <SEP> 5.0
<tb> Water <SEP>&<SEP> preservatives <SEP> QSP <SEP> 100g

<tb> Example <SEP> N 2 <SEP> - <SEP><SEP> Body Lotion <SEP> Deodorant <SEP> and <SEP> Lightening
<tb> Crillet <SEP> 3 <SEP> 2,5
<tb> Novol <SEP> 0.9
<tb> Fluilan <SEP> 2,5
<tb> Carbopol <SEP> 940 <SEP> 0.3
<tb> Bee <SEP> wax <SEP> 2.0
<tb> Triethanolamine <SEP> 0.1
<tb> Glycerine <SEP> 5.0
<tb> Tyramine <SEP> 3.0
<tb> Water <SEP>&<SEP> preservatives <SEP> QSP <SEP> 100g

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 The following examples will demonstrate the lightening effect of tyramine. Example N 3 - Inhibition of human tyrosinase (in vitro) This example demonstrates a dopamine oxidase activity.

Normal human melanocytes (MNH) in culture are lysed by an Igepal 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 not containing (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 L-Dopa. The reaction is initiated by the addition of L-DOPA.

The kinetics are monitored under a wavelength of 490 nm every 10 minutes between TO and 1H then every 30 minutes up to 2 hours.

r+_10' + 20' +30' +40' +50' +60' +90' +120 --------------------------------------------------------------- .____ -_14;1_._.- _-13,4 - - _14,2-_.__ -_14;0 ____ __13,2 _--__ Tyramine 0,1% --------------------------------------------------- ..---------------------------------------------- 5 _ ___ _3-1;7 ._ -__ "_____ Tyramine 0,3% ,--_._--__-_35,g-_-__ --.4 -__- _50,9-__- _52,g-_._ --5-------------------------- 3.g Tyramine 1,0% -27,p Ces résultats démontrent une baisse importante de l'activité de la tyrosinase en présence de tyramine. Comme la présence ou l'absence de cette tyramine est la seule variable de l'expérimentation décrite, il est clair que l'effet observé ne peut provenir que de la tyramine. Enfin, cet effet est spécifique du système observé avec la tyramine puisque l'effet observé est clairement concentration-dépendant. The following table summarizes the average results of the percentages of inhibition, observed compared to the control at the same time, obtained on 8 different tests.

r + _10 '+ 20' +30 '+40' +50 '+60' +90 '+120 -------------------------- ------------------------------------- .____ -_14; 1 _._.- _-13 , 4 - - _ 14,2 -_.__ -_14; 0 ____ __13,2 _ - __ Tyramine 0,1% ---------------------- ----------------------------- ..------------------- --------------------------- 5 _ ___ _3-1; 7 ._ -__ "_____ Tyramine 0.3%, --_ ## STR2 ##, ## STR2 ## These results demonstrate a significant decrease in tyrosinase activity in the presence of tyramine. this tyramine is the only variable of the experiment described, it is clear that the observed effect can only come from tyramine Finally, this effect is specific to the system observed with tyramine since the observed effect is clearly concentration-dependent .

It is understood that we have validated this test also by ensuring the stability of the system by the realization of cases without L-Dopa and without tyramine.

Example N 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 large variations due to interindividual variability. donors.

For this reason, although the previous results leave no doubt about the inhibitory effect of tyramine on tyrosinase, we performed different experiments with another model.

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 The cells used in this example are those of B16 melanomas which produce a lot of melanin and which are commonly used to study the activity of tyrosinase as well as that of melanin synthesis.

These cells are reseeded in 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. concentrations of 0.01 and 0.03% (w / w) or not containing (controls).

A positive control was performed using 0.01 × 10 -3% hydroquinone.

Moyennes des pourcentages d'inhibition, observés enfin d'incubation (48 H) par rapport au contrôle au même temps (S essais différents) Tyramine 0,01% -22,1 --------------------------- ------------------------------------ ------------------------------------------- ----------------- Tyrarnine 0,03% -29,9 fHydroquinone à 0,01.10-%. -20,0 Ces résultats démontrent une baisse importante de l'activité de la tyrosinase en présence de tyramine qui est meilleure que celle obtenue avec l'hydroquinone à concentration comparable. Comme la présence ou l'absence de cette tyramine est la seule variable de l'expérimentation décrite, il est clair que l'effet observé ne peut provenir que de la tyramine. Enfin, bien que réalisé avec seulement 2 concentrations différentes de tyramine, cet effet est spécifique du système observé avec la tyramine puisque l'effet observé est concentration-dépendant. As before, the cells are then lysed by an Igepal buffer associated with ultrasound, the activity of tyrosinase is quantified and standardized with respect to the amount of proteins present.

Mean percent inhibition, observed finally incubation (48 H) compared to control at the same time (S different tests) Tyramine 0.01% -22.1 ------------- -------------- ------------------------------------ ------------------------------------------- ------- ---------- Tyrarnine 0.03% -29.9 fHydroquinone 0.01.10-%. These results demonstrate a significant decrease in the tyrosinase activity in the presence of tyramine, which is better than that obtained with hydroquinone at comparable concentration. Since the presence or absence of this tyramine is the only variable of the experiment described, it is clear that the observed effect 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 observed effect is concentration-dependent.

Example N 5 - Inhibition of melanin synthesis (in vitro) Normal human melanocytes (NINH) in culture (confluence between 60 and 80%) are placed in the presence of buffer of a buffer containing tyramine, at concentrations of 0.003 and 0.01% or not containing (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 5 times to UV-B (20mJ / cm 2) while the other half do not receive irradiation (controls).

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

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

Cellules non irradiées Cellules irradiées Tyrnmine 0,003 10 -30 -28 ---------------------------------------------------------- ----------------------------- ---------------- ---------- Tyramine 0,01 % -32 -32 Arbutine 0,09 % - 11 - 8 Ces résultats démontrent: # d'une part, qu'en l'absence des irradiations par les U V13, la tyramine diminue le taux basal, ce qui traduit une diminution de la quantité totale de mélanine formée d'où un effet blanchissant attendu, # d'autre part, qu'après les 5 irradiations par les UV-B, l'inhibition de la synthèse de la mélanine en présence de tyramine est maintenue plus basse que chez les témoins, limitant ainsi l'effet inducteur du rayonnement sur la néosynthèse (facteur *2 dans le cas des témoins). The following table summarizes the average results of the percentages of inhibition, observed compared to the controls at the same time, obtained on 5 different tests.

Non-irradiated cells Irradiated cells Tyrnmine 0.003 10 -30 -28 -------------------------------------- -------------------- ----------------------------- - --------------- ---------- Tyramine 0.01% -32 -32 Arbutin 0.09% - 11 - 8 These results demonstrate: # of on the one hand, in the absence of U V13 irradiations, tyramine decreases the basal level, which reflects a decrease in the total amount of melanin formed resulting in an expected whitening effect, # on the other hand, that after the UV-B irradiations, the inhibition of melanine synthesis in the presence of tyramine is kept lower than in the controls, thus limiting the inducing effect of the radiation on the neosynthesis (factor * 2 in the witnesses).

Since the presence or absence of this tyramine is the only variable of the experiment described, it is clear that the observed effect 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 observed effect is concentration-dependent.

 Example N 6 - "Stain-repellent" effect (in vivo) This test was carried out on 20 female volunteers aged 45 to 65 years with age spots on the skin of the hands.

After precise identification and measurement of the size and staining of 5 spots per hands and by volunteers, the protocol consists of applying the product to be tested incorporated (5%) in the cream described in the example N 1 during a period of a month and a half on one hand (randomly drawn), the other serving as a control (thus treated with the same cream but not containing tyramine). Of course, skin staining measurements of 5 spotless areas were also performed to allow standardization of the results.

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 At times + 1 month and + 1 month and a half, quantification of the cutaneous coloration is 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 age spots staining of the treated hands were respectively 6.4% and 24% lower than those treated with the cream not containing 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 and a real effectiveness on a less dermal coloration during exposure to natural or artificial UV The concentration of tyramine may 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. be limiting.

It is possible to incorporate tyramine 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.

Tyramine can be combined in cosmetic compositions with any other ingredient normally used in cosmetics: extraction and / or synthetic lipids, gelling and viscosity polymers, surfactants and emulsifiers, hydrosoluble active ingredients, extracted from other plants, tissue extracts, marine extracts.

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

These cosmetic or dermopharmaceutical compositions are used for the preparation of medicaments intended for the care of the skin, particularly its

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 whitening and its lesser color when exposed to natural or artificial UV.

Tyramine is used alone or incorporated in cosmetic or dermopharmaceutical 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, wools and any materials that can be used to make clothes and underwear during the day or night, directly in contact with the skin or hair to allow continuous topical delivery.

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Claims (10)

 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 may: come from plant extract, preferably from 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 supports including liposomes, # be grafted, on the OH or on the NHz, by any linear or branched alkyl chain, 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 dosage form used in cosmetics or dermopharmacy: O / W emulsions and E / H, 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 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. 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 viscosity polymers, surfactants and emulsifiers, hydro- or liposoluble 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 sun, after-shave products, hair removal creams or post - depilatories. <Desc / Clms Page number 10>  8. Cosmetic or dermopharmaceutical composition according to one of claims 2 to 7, for the care of the skin, particularly its whitening and brightening and for a less dermal coloration during exposure to natural or artificial UV. 9. Use of the single 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 for the care of the skin, particularly its whitening and its less staining. when exposed to natural or artificial UV. 10. Use of the single tyramine according to claim No. 1 or incorporated in cosmetic or dermopharmaceutical compositions according to claims 2 to 7, 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, wools and any materials that can be used to make clothes and underwear during the day or night, directly in contact with the skin or hair to allow continuous topical delivery.