FR2756183A1 - Synergistic composition containing argan oil and argan peptide - Google Patents

Abstract

Pharmaceutical or cosmetic compositions contain a combination of argan oil (AO) and argan peptides (AP). Argan, or argan-tree, (Argania spinosa L.) is a plant of the Sapotaceae family, found in Morocco.

Description

The present invention relates to the use of a synergistic combination of oil and Argan peptides and its application for preventing and / or treating disorders related to skin aging, especially during menopause.

More particularly, the invention relates to a dermocosmetic composition, characterized in that it contains as active ingredient at least one combination of oil and Argan peptides.

It may further contain: - pharmaceutically acceptable excipients suitable for
oral or topical administration; other active substances and / or fractions of oil and / or peptides
of Argan, such as vitamin E and phytosterols - an oil concentration of between 0.01% and 20% by weight and
a peptide concentration of between 0.001% and 10% in
weight, relative to the total weight of the composition;
The invention is also directed to the use of a composition for the preparation of medicaments for the treatment of cutaneous aging, including menopausal-related disorders; such a composition may, in addition, contain at least one other active ingredient, in particular hormones, such as estrogens.

Finally, the invention relates to a cosmetic treatment method for improving the appearance of the skin and / or superficial body growths by applying the above-mentioned compositions locally, in particular to reduce menopausal disorders.

The senescent skin is characterized by a certain number of alterations, the principal one being a cutaneous atrophy following the slowing down of the proliferative activities.

During skin aging, there is a flattening, including a decrease in thickness of the epidermal layer, a deep disorganization of the extracellular dermal matrix with a decrease in the proteoglycan content, a modification and a rarefaction of the support fibers.

At the macroscopic level, these changes result in a dry skin, an accentuation of the wrinkles, a loss of elasticity and a tarnishing of the complexion.

Menopause will speed up this process. Indeed, the skin is a hormone-dependent organ, since there are cutaneous receptors with sex steroid hormones.

Thus hormone replacement therapy has a beneficial effect on the skin of menopausal women.

Argan or Argan (Argania spinosa L) is the most characteristic plant species of Morocco. The plant belongs to the botanical family of
Sapotaceae. He is a survivor of the tropical tertiary flora that has adapted well to the North African climate.

The earliest reference to the argan tree dates back to 1219 and was made by the doctor Ibn Al Beithar in a "Treaty of the simple". The first botanical description was given by Linne in 1737 who named it Sideroxylon spinosum (ironwood).

Argania spinosa is a tree that can reach 8 to 10 meters in height.

The trunk is short: 2 to 3 mothers, multiple and tormented. The twigs are thorny which ensures some defense of the tree. The leaves are alternate, lanceolate, 2 to 3 cm long, they have a green color, dark on the upper side and are lighter underneath. They are sub-existent. In times of severe drought, the tree completely loses its leaves which reappear after the return of the rains.

The flowers are pentameric with sepals white pubescent. The corolla is yellow with 5 stamens. The ovary is superior, pubescent. The fruit is a berry with 2 or 3 seeds welded in a false drupe which, at maturity, has the size of a yellow walnut veined with red. The pulp covers a very hard core (Argan nuts). This walnut contains 3 almonds albuminées and oily representing approximately 3% of the weight of the fruit and containing 50 to 60% of oil.

The Argan tree is located mainly in southwestern Morocco, but there are some isolated stands in the north-east of the country in the Beni Snassen mountain range and in the center.

The argan plantation currently covers 822 000 ha according to the Direction des Eaux et Forêts (1989). Currently, most of the argan forest covers a coastal strip from the mouth of Oued Tennsilf to that of Oued Souss.

The Argan tree is valued in different forms by the Moroccan populations. First and foremost, the particularly hard wood is mainly used as firewood and the resulting coal supply.

But the most interesting applications originate from the kernel of the seed. This one provides an oil, then in a second time, a cake.

The preparation of argan oil is carried out by the local population in a traditional way. It is extracted by breaking the stone of the fruit between two stones. The seeds are roasted on soft fires. The chilled almonds are ground in a hand mill and turned into flour, which is sprayed with hot water and kneaded by hand. The oil floats on the surface and is recovered as and when.

After extraction of the oil, the cake is collected and then pressed by hand into more or less ovoid loaves, of a brownish or blackish color and the very bitter taste.

In an industrial manner, Argan oil is obtained by pressing to obtain the virgin oil, or by extraction with an apolar solvent such as hexane.

Cakes are preferably obtained virgin oils, by pressing, so as not to be polluted with hexane or the residual solvent.

This virgin oil can be enriched by addition of vitamin E and / or phytosterols. Argan oil is composed of glycerides of fatty acids and an unsaponifiable part.

Glycerides
Glyceride analysis indicates that Argan oil contains more than 96% by weight triglycerides, 1.5% diglycerides and free fatty acids, 0.3% monoglycerides.

Fatty acids:
The fatty acid analysis of this oil shows a predominance of C 18-1 and C 18-2 acids, including oleic acid (45%) and linoleic acid (35%). The rather high content of polyunsaturated fatty acids represents, from the point of view of food, a certain interest.

On the other hand, the virtual absence of linolenic acid means that this oil is unlikely to oxidize or polymerize during storage. The other fatty acids contained in important proportions are: C 16 palmitic acid (15%) and C 18 stearic acid (6%).

The unsaponifiable rate of argan oil is on average 1%. The composition varies according to the authors. The most recent studies describe 37% hydrocarbons and carotenes. 7.5% tocopherols, 20% sterols and triterpenic alcohols, 6% xanthophylls and 9.5% miscellaneous constituents.

The most active molecules are tocopherols which have very strong anti-radical activities, but also sterols. The latter regulate the permeability and fluidity of cell membranes and thus, all existing exchanges between the cell and the outside world, that is to say its direct environment. While less important than tocopherols, sterols also have antiradical properties.

The protein part: the Argan cake is composed essentially of proteins, 25 to 35% depending on the provenances of the cake, non-extractable residual lipids: 20 to 27% of water, the content of which varies according to the drying state of the product. at 25%, polysaccharides 15 to 25% including 2 to 5% cellulose, ash 2 to 5% and trace elements such as phosphorus, calcium, potassium and sodium.

Proteins are the most abundant molecules. In terms of protein, Argan cakes are comparable to those of soy or peanut. Local people use these cakes for animal feed. No cases of toxicity due to their chemical composition have been noticed. Argan proteins can be extracted with aqueous, alkaline or neutral solvents, but the proteins in solution are not stable and precipitate. In addition, they are devoid of biological properties, apart, of course, their nutritional aspect. In order to solubilize them, these proteins can undergo chemical or enzymatic hydrolysis. The latter is preferred because it is more specific and less denaturing.

The enzymatic hydrolysis is conducted by contacting the freshly extracted proteins with an aqueous solvent or simply still in the state of the cake in water with a protease. This enzyme then hydrolyzes the proteins into peptides of molecular weight less than 20,000 daltons.

These peptides are then soluble in water and thus recovered after a solid / liquid separation in the aqueous phase. They are biologically active as demonstrated by the work reported below.

Taken alone, Argan's oil and peptides have anti-aging activities that have been quantified in cell culture, animal pharmacology and human clinical trials.

The oil intervenes on the principal parameters of the aging that are: - the diminution of the cellular activity, - the perturbation of the cellular exchanges and the accumulation of toxins, - the diminution of the quality of the intercellular cement, - the decrease of the contributions nutritious at the cellular level.

1) Proliferative activity leading to a revival of
cell activity
The stimulating activity of the enriched oil at a concentration of 0.003
% has been demonstrated on fibroblast culture (line L 929) in
using the XTT method described later in more detail.

2) Anti-radical activity
It was evaluated in vitro by the DPPH test. Concentration
inhibitor of oil required to halve the rate of
free radicals (IC 50) obtained is 1.3 mg / ml.

3) Restructuring and moisturizing effect
1l was objectified on animals deficient in AGE
The treatment of these animals with Argan oil decreases
quickly the perspiration in the animal deficient in AGE and this
for a long time (activity from the 4th day of application). he
it is a real reconstruction of the integrity of the stratum corneum,
because a paraffin oil turns out to be inactive on this model.

 This moisturizing effect has been confirmed in the human clinic.

4) Nutritional effect
Argan oil has emollient properties due to its oil quality.

However, its action is clearly different from that of a
classic emollient by its particular composition (fatty acids
unsaturated, etc.).

Peptides are mainly involved in aging disorders related to alterations in cellular metabolism: a drop in oxygen consumption, a decrease in collagen synthesis, and so on.

1) Proliferative activity
It has been evaluated on several cell lines by the method of
XTT (with reading at To + 48 h).

The stimulating activity of the peptides 0.1% to 0.3% and
1% was objectified at the same time on fibroblastic line and on line
Epithelial.

This property of cellular stimulation was confirmed by a test
in vivo in animal pharmacology, by monitoring the evolution of the index
in guinea pigs after daily application for two
weeks, of a preparation containing 1% of Argan peptides.

2) Oxygraphy
This test was performed in vitro on fibroblast culture. Has
concentrations of 0.1% and 0.3% in peptides oxygenation on
fibroblast culture revealed considerable improvements in the
rate of oxygen consumption.

3) Anti-collagenase activity
Argan peptides protect collagen fibers from attacks
enzyme. This has been objectified in vitro on agarose gel. The IC 50
obtained with the Argan peptides is 0.1% and the IC 100 is 0.5%.

In the context of the present invention, fortuitously, a potentiation of the proliferative activity of the peptides by argan oil has been demonstrated. This property has been demonstrated in vitro on murine fibroblast culture (line L 929). The principle of the test is based on the enzymatic transformation of a tetrazolium salt, XTT into a colored product: formazan.

XTT is reduced by the mitochondrial dehydrogenases of living cells in the presence of an electron coupling agent, coenzyme Q to a water-soluble yellow / orange compound, formazan measurable by spectrophotometry at 450 nm.

At 6 pm, the cells are seeded with S.V.F.

A to, the culture medium is removed and the active agents (peptides and / or oil) are introduced at different concentrations (dilution in the culture medium).

At 48 h, a solution of XTT is introduced in the presence of coenzyme Q,
After 3 hours of incubation, the metabolism of XTT is stopped by adding an SDS solution.

Huile enrichie Peptides Peptides(1 %)
(0,003%) (1%) + Huite
enrichie(0,003%)

The reading of the optical density is carried out at 450 nm. It is proportional to the living cell population. The percentage of activity is calculated as
DO control - DO processed
xx 100
DO witness
The results are shown in the graph of the attached figure and in the table below:
Potentiation of Peptide Activity by Enriched Oil% Activity / Witness SVF

Peptides Peptides Enriched Oil (1%)
(0.00%) (1%) + Huite
enriched (0.003%)

<tb> Oil <SEP> Enriched <SEP> (0.003%) <SEP> 6%
<tb> Peptides <SEP> (1%) <SEP> 76%
<tb> Peptides <SEP> (1%) <SEP> + <SEP> Enriched <SEP> Oil (0.003%) <SEP> 96%
<Tb>
It is thus observed that Argan oil has been found to potentiate the stimulating activity of Argan peptides on cell culture. This synergy of action, as well as the complementarities of these two active ingredients on the other aging parameters, make this combination an effective treatment adapted to the fight against skin aging.

The above properties are advantageously used when using this combination or compositions containing it, for the preparation of a medicament for the treatment of skin disorders associated with aging.

These compositions may be of cosmetic or pharmaceutical use, in the form of creams, emulsions, lotions, masks, or even capsules by the oral route, examples of which are given below.

In particular, they can be combined with compositions which are useful in particular as medicaments, such as hormones and in particular estrogens, in the treatment of cutaneous disorders of the menopause.

EXAMPLE 1: Moisturizing day cream based makeup
Argan oil enriched 0.5 to 2
Argan peptides 0.03 to 2
0.2% to 0.5% pyrrolidone carboxylic acid
Marine collagen hydrolyzate 1 to 3
Hydrogenated castor oil, ethoxylated 1 to 5
Fluid paraffin oil 1 to 5
Cyclopentasiloxane oil 5 to 12
Squalane 1 to 5
0.1% to 0.5% carboxyvinyl polymer
Triethanolamine 0.2 to 0.5%
Glycerin 2 to 4
Conservative qs

Perfume q.s.

Purified water qs 100 g
EXAMPLE 2: Cleansing milk for dry skin
Argan oil enriched 0.5 to 1
Argan peptides 0.02 to 1
Sorbitan monopalmitate 0.5 to 2
Polyethoxylated sorbitan monopalmitate 3 to 6
Polysorbate 20 1 to 3
Liquid paraffin 8 to 15
Capric / caprylic triglycerides 1 to 2
Butanediol 1.3 2 to 4
0.2% to 0.5% carboxyvinyl polymer
Triethanolamine qs

Conservative q.s.

Perfume q.s.

Purified water qs 100 g
EXAMPLE 3 Nutritive cream for very dry skin
Argan oil enriched 0.5 to 3
Argan peptides 0.05 to 2
Polysaccharide derivative 0.1 to 4
Stearic acid 1 to 3
Polyoxyethylene glycol stearate 2 to 6
Vegetable wax 1 to 3
Synthetic whale white 1 to 2
Jojoba oil 2 to 8
Glycerine 2 to 5
Conservative qs

Perfume q.s.

Purified water qs 100 g
EXAMPLE 4 Anti-wrinkle concentrate
Argan oil enriched 0.1 to 3
Argan peptides 0.01 to Q05%
Palmitate vitamin A 0.1 to 1
Glycerol Ester of Vitamin F 0.1 to 1
Safflower oil qs 100g
EXAMPLE 5 Anti-Wrinkle Concentrate
Argan oil enriched 0.1 to 3 ù
Argan peptides 0.01 to 0.054%
Unsaponifiable Argan 0.01 to 0.5%
Vitamin A palmitate 0.1 to 1
Purcellin oil qs 100 g
EXAMPLE 6 Anti-Wrinkle Mask
Argan oil enriched Q5 to 4
Argan peptides 0.01 to 2
Vegetable proteins 1 to 5
Hyaluronic acid salt Na) 0.01 to 0.2%
Sucrose Stearate 1 to 3 Sucrose Distearate 2 to 5
Capric and caprylic triglycerides 2-8
Liquid paraffin 5 to 12
Polyoxyethylene glycol 600 2 to 10
Carboxyvinyl polymer 0.2 to 1
Triethanolamine 0.5 to 1
Perfume qs

Conservatives q.s.

Purified water q.s. 100 g

Claims (8)

 Argan peptides.  contains as active ingredient at least one combination of oil and  CLAIMS 1) Pharmaceutical or cosmetic composition, characterized in that it 2) Composition according to claim 1, characterized in that it contains  pharmaceutically acceptable excipients suitable for  oral or topical administration. 3) Dermo-cosmetic composition according to claim 2, characterized in  what it is, in addition, enriched by other substances and / or fractions  oil and / or Argan peptides, such as vitamin E and  phytosterols. 4) Composition according to one of claims 1 to 3, characterized in that  the oil concentration is between 0.01% and 20% by weight and the  peptide concentration is between 0.001% and 10% by weight,  relative to the total weight of the composition. 5) Use of a composition according to one of claims 1 to 4 for the  preparation of drugs for the treatment of aging  cutaneous, including troubled menopause. 6) Use according to claim 5, characterized in that the  composition contains, in addition, at least one other active ingredient, in  hormones, such as estrogens. 7) Cosmetic treatment method for improving the appearance of the skin  and / or superficial body growths, characterized in that a  Composition according to one of Claims 1 to 4. 8) Process according to claim 7, characterized in that the  disorders related to menopause.