FR2765481A1 - Vitamin B6 amino lipid complex - Google Patents

Abstract

A vitamin-amino lipid complex (ALV) comprising the precursors for the biosynthesis of ceramides and containing serine or its precursor, vitamin B6, vegetable oil and palmitic acid, is new. Also claimed are : (A) cosmetics containing 0.5 - 10% of ALV; and (B) a method of in vivo synthesis of epidermal lipids by applying ALV to the skin.

Description

-1 -

Vitamin amino-lipid complex.

  The present invention relates to a particular cosmetic composition hereinafter called "amino-lipid vitamin complex", which stimulates the in situ synthesis of epidermal lipids and thus increases their level in the stratum corneum. It also relates to its use in cosmetic forms applied directly to the skin, such as cream, milk, gel, etc., for the purpose of

  cosmetic, physiological or dermatological treatment of the skin.

  It is well known nowadays that the lipids synthesized in the cells of the stratum corneum or coméocytes, then excreted in the extracellular space, play an important role in the cohesion of said cells between them (and, by contrast, for their desquamation) , in the fight against the penetration into the layers of the epidermis of foreign substances (or permeability of the stratum cornea), and the maintenance of hydration of the epidermis (or regulation of cutaneous water flows). These epidermal lipids represent 10 to 12% of the total weight of the dry epidermis. They are of different types (derived from cholesterol, ceramide, cerebroside, phospholipids, triglycerides, diglycerides and fatty acids), some playing a more important role than others, in the above phenomena. Epidermal lipids, with the exception of phospholipids and glycerides, are also called cohesion lipids (ceramides, cerebroside, cholesterol, cholesterol sulfate). Indeed, cholesterol in the form of cholesterol sulfate in the presence of calcium increases cohesion between cells; on the other hand, in its non-sulfated form, it allows the desquamation of coméocytes. Cholesterol sulfate has also been shown to prevent the development of skin carcinomas (cancer of the epithelia) in the early stages (promotion), (Cancer res., 55: 4865-9, 1995 Nov. 1). In addition, during the modification of the skin barrier, by a solvent for example, it has been demonstrated the essential role of cholesterol for the restoration of the barrier function of the skin. As far as ceramides are concerned, seven different types have been identified to date. The relative proportion of one type compared to others in the epidermis depends on various parameters, such as age, skin condition, type and intensity 2 - of the aggressions suffered by the skin. In the case of normal skin, the epidermal cells resynthesize the type of ceramide which is deficient. However, under certain conditions, and in particular with age, it has been shown that the ability to resynthesize lipids by epidermal cells decreases. Many cosmetologists have tried to provide the skin with ceramides of different types. Some people think that type 2 improves the skin, others that it increases with age and that on the contrary type 3 is more indicated. Finally, there are studies also showing that ceramides could have a detoxification role.

  by complexing certain very reactive lipids (sphingosines).

  Many cosmetics containing ceramides are already available on the market. Several French and foreign patents published also reveal the use of ceramides without necessarily dissociating the type, in cosmetic compositions. However, it is important to underline that the skin, to function well, does not need exclusively ceramides but of all the epidermal lipids. In addition, any exaggerated intake of one type of lipid or another may cause an imbalance in the natural levels of the various lipids in the skin and / or worsen said existing imbalance, in particular, direct application to the skin, Cholesterol sulfate provides a significant condition of scaly skin (ichthyosis). It therefore appears more interesting to provide the skin with the elements it uses to re-synthesize its epidermal lipids according to its needs, rather than imposing or unbalancing it by an exogenous supply of a type of lipid rather than of another. This concept of stimulating epidermal cells has already been used by the invention described in document WO-94/23694. According to this document, the application of one or more precursors of the biosynthesis of ceramides on the skin makes it possible to increase the level of ceramide in the epidermis. However, the use of sphingosine or any other sphingosine precursor such as tetraacetyl phytosphingosine allows the in situ increase of the level of ceramides only, but has the disadvantage, as the chemical study shows, of having little effect on the level of the cholesterol / cholesterol sulfate pair and on that of fatty acids. However, the function fulfilled by each of the epidermal lipids is specific, so that any excess of one type of lipid (example: ceramide), compared to another does not make it possible to compensate for the deficiency of another type

  epidermal lipid (other than ceramide).

  The objective of the invention is therefore to remedy this drawback by proposing a vitamin-rich amino lipid complex which, used alone or in a cosmetic formulation for topical application, allows the in-situ increase of all epidermal lipids. This amino-lipid complex vitamin which will be designated most often under the name of "complex ALV" for the sake of brevity, in the remainder of this presentation, is remarkable by the fact that it consists mainly of the following 4 elements , namely: - serine or a precursor of the biosynthesis of serine; - palmitic acid; - vitamin B6;

- vegetable oil.

  Thanks to the precursor of serine, the neosynthesis of serine takes place in situ, so that the synthesized serine is used by the cell as a substrate for the biosynthesis of ceramides. Vitamin B6 also plays a role in

  biosynthesis of ceramides as a cofactor for one of these reactions.

  However, it is thanks to the complex formed by the four elements of said amino-lipid vitamin complex: serine, vitamin B6, palmitic acid and vegetable oil, that can be carried out surprisingly after its application on the skin, in addition to the neosynthesis of ceramides, that of all epidermal lipids, by skin cells. In addition, after the skin application of the amino-lipid vitamin complex according to the invention, a sharp increase in the synthesis of cholesterol and cholesterol sulfate is observed, and

cerebroside.

  The cutaneous application of this ALV complex therefore has the major advantage of providing the substrates necessary for the cell to synthesize all the epidermal lipids. However, the cell will specifically produce only those which it needs. The skin thus compensates for its lipid deficiencies due to various parameters, such as the environmental and physiological conditions, in which it is found. By reconstituting the epidermal lipids at the level of comeocytes, the skin keeps its hydration, its "good health" and its beauty. Thanks to such adapted and specific stimulation, no imbalance in the adequate levels of epidermal lipids present in the stratum corneum is caused. In addition, this amino-lipid vitamin complex can be added to all conventional cosmetic formulations for topical use, and thus be

  easy to apply and use and better storage.

  The invention also relates to a method of treating dry and / or sensitive, and / or atopic, and / or aged skin, according to which said vitamin-amino-lipid complex, alone or inserted in any formulation, is applied to the person's skin. cosmetically acceptable. By allowing the synthesis of different epidermal lipids, this amino-lipid vitamin complex makes it possible, in fact, to fight against the signs of aging, namely in particular the loss of suppleness of the skin linked to a reduction in skin hydration, and regulates furthermore the desquamation of the surface layers of the epidermis. It also prevents the appearance of skin carcinomas, in particular during direct contact

  of the skin with a carcinogenic promoter (12-0-tetradecanoylphorbol13-

  acetate or any other promoter) by allowing the synthesis of the cholesterol / cholesterol sulfate couple at the level of the epidermal cells. The use of said ALV complex can also be recommended as a preventive of said

skin cancer.

  The goals, features and advantages above and others will emerge

better from the following description.

  The amino-lipid complex vitamin according to the invention, comprises in the most simple way, at least one precursor of the biosynthesis of serine or serine

  itself, vitamin B6, at least one palmitic acid and vegetable oil.

  The precursors of serine can be phosphoserine, 5-hydroxypyruvic acid, glycine. These precursors give, thanks to cellular machinery, serine according to the specific reactions below: Phosphoserine + H2O phosphatase> Phosphate + serine Hydroxy-pyruvic acid transaminase> serine Glycine <serine hydroxymethyl tranferase> serine (by interconversion) Serine and vitamin B6 are elements necessary for the synthesis of all types of ceramide as shown in the reaction scheme below; serine being a precursor of ceramides and vitamin B6 playing a role of cofactor in the reaction between serine and palmitoylcoenzyme A. The latter is the product of the reaction between a palmitic acid and a coenzyme A. CH (CHj1rFOOH + CoA Sphinganine CIzOH Palmitic Acid HC-NH

ADF I I

] .5 CIiCH, CHzOH Cl I carboxylase

O0 1I C-H CH

H C-NH2 I I

  CH2ICHI14C-S-CoA Palmitoyl CoA H- -OH + i O C

CH2OH I CH, CH2

I C- H I

H - -NH2 L-Serine C CH2 CH2

COOH CH, 2 I

  synthetase I CH, CH2 CoA._. , Vltamine B6 dependent CH2 I I CoA CH, CHz Có, OH CHz I HII C --- NHz Dehydrosphinganine CH CH2 - CH2 Sphingasine I I

Cç 1I CH2 CH,

CHz I I

CH3 CH2 I IC

  NADPHq'IICIi H NAPH9 Ré & dudase I CCH2, JCHC-S-CoA PalmitoyI-CoA CH2 CH, NADPH Reduclase CH2 CjH

NADP I CH2 CH2

  CH20H CH2 Acetyl CoA II carboxylas CH CH, H- C-NH2 Sphinganine CHz carboxylase CH2 HC-OH CHz CH, Il CH2 CH '{CH2) 14COOH + CoA JI [C Hlu14 Palmitic acid CH, CH] CH2 CH2 (or fatty acid CH 31 essentlel. Ceramide

CH2

  CH3 -6- Since coenzyme A is regenerated at the next stage of the biosynthesis of ceramides, it is therefore not necessary to supply it to the cell with this vitamin-amino-lipid complex. Palmitic acid, meanwhile, is provided by the amino-lipid vitamin complex. Vegetable oil composed of unsaturated fatty acids plays a role in the synthesis of all epidermal lipids as well as in the neosynthesis of ceramides. These unsaturated fatty acids can be

supplied by any vegetable oil.

  Advantageously, the vegetable oil is blackcurrant seed oil which additionally contains essential unsaturated fatty acids of palmitic acid. The essential fatty acids necessary for the synthesis, in particular of ceramides to

  from sphingosine are provided by this oil.

  It should be noted that palmitic acid, like the unsaturated fatty acids provided by vegetable oil, are not exclusive precursors of ceramides and that the reactions illustrated above are not the only ones

  stimulated by the 4 elements present in this amino-lipid vitamin complex.

  Indeed, after application of the latter on the skin all types of lipids are

  synthesized by the cells of the stratum corneum according to their need.

  To obtain the expected results, the proportion of the compounds of the said amino-lipid vitamin complex must be between: palmitic acid ........................... ............................. 0.5 to 40% serine or one of its precursors ......... ......................... 3 to 30% vegetable oil of blackcurrant seeds ............... .............. 40 to 80% vitamin B6 .............................. .........................

  3 to 10% Preferably, the percentages are palmitic acid ..................................... ........................... .DTD: .. 1% serine or one of its precursors .......... ......................... 16% vegetable oil of blackcurrant seeds ................................. .DTD: .. 77% vitamin B6 ................................................ DTD: ........................ 6% -7- Said ALV complex can be applied directly to the skin, after shaking, .. DTD: by touch or by light massage.

  However, a physiologically acceptable vehicle (physiological pH 7) can be added to the said complex so that the latter is applied to the skin at an adequate concentration. The role of the vehicle or excipient is to facilitate the application of said complex. It generally conditions the type of final product (emulsion, gel, lotion ...). The vehicle can be an O / W, W / O emulsion or even a gel if the gelling agent is self-emulsifying (Permulen TR1 (Trademark), Permulen TR2 (Trademark)). The vehicle is usually inert or may have other functions in addition. The priority function is to increase the penetration of said complex into the skin, to improve its action in situ. Other compounds can advantageously be added to this amino-lipid vitamin complex such as vitamins, for example vitamin A for its beneficial functions on skin cells (proliferation regulator, anti-aging ...), vitamin E for its antioxidant role, as well as of course preservatives, trace elements (Mg and Zn), enzyme inhibitors (such as 5a reductase inhibitor), as well as other fatty acids, fatty acid derivatives or products allowing the dispersion of the amino-lipid vitamin complex and the formation of liposomes. The emulsifiers are chosen to give emulsions in the form

O / W or E / O.

  An interesting formulation of the final composition of the product comprising the amino-lipid vitamin complex according to the invention is an emulsion in

  which the oily phase corresponds to the oily part of said amino complex

  vitamin lipid (vegetable oil and palmitic acid), the aqueous phase in its aqueous part (i.e. vitamin B6 and serine). These two phases are held together thanks to emulsifiers, it is well known that the emulsifier is chosen according to its hydrophilic lipid or HLB balance, and thus determines whether the final product is in the form of an oil-in-water emulsion ( HIE) or water in oil (W / O). This amino-lipid complex vitamin can also be added to conventional cosmetic formulations, type oil in water I water in oil / cream / milk I lotion I gel, at an adequate rate -8 - allowing its action to know a rate between 0; 5 and 10% and

  preferably between 1 and 5%. It advantageously constitutes 3% of the final product.

  The final packaging of the product is often related to the formulation of the latter, so for example, when the composition of the final product is in the form of a cream (O / W or W / O emulsion) the latter is stored in a small

  non-deformable container (pot) or deformable container (like tubes).

  By way of nonlimiting example, the invention can be illustrated by the examples of formulations of cosmetic products containing the amino-lipid complex.

vitamin according to the invention.

  When the final product is in the form of an oil-in-water cream, such a cream can be formulated as follows: Sorbitan monostearate ................ .................................................

  .......... 3.26% Cetyl phosphate + cethanol phosphate diethanolamine ............................ .2.72% Isocetyl stearate .... .DTD: .................................. ....................................... DTD: .. 4.00% Seed oil blackcurrant ...................................... DTD: ........ .............................. 0.80% Palmitic acid ........... DTD:. .................................................. ...................... DTD: ..0.20% Dicaprylate / Propylene glycol dicaprate .............. ....... DTD: ........................... 4.00% Polyethylene glycol 1500 ....... ..DTD: .............................................. .................... 1, 00% Triglycerides of saturated fatty acids .................... .............. .DTD: ........................... 4.00% Copolymer of acrylic acid ....... DTD: ..................................... ................. 0.25% Vitamin B6 ........................... ............................................. .DTD: ..................... 0.06% Triethanolamine 98-99% .................... DTD: ............................................... ........ 0.25% Serine ..................................... ............................... .DTD: ................ ...................... 0.16% Water / preservatives .......... DTD: ......... .................................................. ... qs 100.00 9- The second example of a cosmetic formulation containing the ALV..DTD complex: according to the invention is a water-in-oil emulsion in the form of a cream.

  Such an emulsion may have the following formula: Decamethylcyclopentasiloxane / Dimethylsiloxane glycol copolymer .......... 18.00% Dimethyl silicone / Trimethylsiloxysilicate ..............

  ................................... 5.00% Decamethylcyclopentasiloxane / aluminum magnesium stearate ...... .......... DTD: ... 1.50% Dimethyl silicone ............................. ................ DTD: ................................ ........... 3.60% Palmitic acid ....... DTD: ........................ ................................................. DTD : ............ 0.40% Blackcurrant seed oil ............................ DTD: ................................................ 2.00% Vitamin B6 ............................................ ................... .DTD: ............................ ........ 0.03% Serine ........................ DTD: ........... .................................................. ............ DTD: ........ 0.03% Sodium chloride ..................... .................. DTD: .............................. ................... 1.00% Water / preservatives ........................ ................................. .DTD: .............. .......... qs 100.00 A 5 alpha reductase inhibitor can advantageously be added to the abovementioned formulations at a rate of between 0.5 and 5%, and preferably at 3%. The third example concerns an emulsion-water in oil type milk which can be formulated as below: Sorbitan mono-oleate / ethoxylated hydrogenated castor oil / hydrogenated castor oil / beeswax ... ............................................. .DTD: .. ................. 4.00% POE hydrogenated castor oil (7) .......... DTD: ................................... ............... 2.00% fluid vaseline oil ........................... ................................ .DTD: ............... .... 10.00% Isopropyl palmitate ............... DTD: ..................... .............................................. 5, 00% Capric / caprylic acid triglycerides ............................ .DTD: ............ ........... 10.00% Blackcurrant seed oil ................. DTD: ........... ................................................ 0, 77% Palmitic acid .............................................. ............ .DTD: .................................. 0 , 01% Vitamin B6 .................... DTD: ....................... ......................................... 0.03% Serine .... .................................................. ..... DTD: ........................................... 0.03% Magnesium sulfate 7H20 ......................................... ........... DTD: .................... 0.70 % Water / preservatives ............................ DTD: ................ ...................................... qs 100.00 -10- .. DTD: The fourth example given concerns an emulsion-oil-in-water milk.

  Such milk can be formulated as below: Sorbitan monostearate ...

  .................................................. ................ 2.82% Isopropyl palmitate .......................... .............. DTD: .................................. ......... 9.00% thick petrolatum oil .................................. ................ .DTD: ........................... 1.00% Cetyl alcohol ............ DTD: .................................. ....................................... DTD: ..... 0.50% Cetyl ether P.OE (20) ............................ DTD: ................. ........................... 2.18% Methyl phenyl polysiloxane ................ ....................................... .DTD: ........ .0.50% Blackcurrant seed oil ................. DTD: ..................... ....................................... 3.00% Palmitic acid ..... ............................................. DTD: ... ....................................... 1.00% Polymer of acrylic acid .. .................................................. ... .DTD: .............. 0.18% Vitamin B6 ......................... ...... DTD: .......................................... ................. 0.30% Serine ............................ ........................................ .DTD: ....... ............................... 0.50% Triethanolamine 98-99% ........... ......................................... .DTD: ...... ............... 0.18% Water / preservatives ........ DTD: .................. .................................................. ..... DTD: .q.s. 100.00 A fifth example of a cosmetic product containing the ALV complex can be given. It can be a liposome cream which can be formulated as below: Cethyl alcohol, polyglycerol ... DTD: ....................... ............................................ 3.80% B- sitosterol .............................................. DTD:. .................................................. 3.80% Diketyl phosphate ........................................... ............ .DTD: ................................ 0.40 % Sunflower oil.............................................. ................... .DTD: ................... 33.00% Blackcurrant seed oil .................................................. .................. DTD: ......... 2.00% Palmitic acid ............... .................. DTD: .............................. .......... 0.05% Polymer of acrylic acid ............................... ........................... .DTD: ............ 0.20% Vitamin B6 .. ........................ DTD: ........................ ........................................ ...... 0.30% Serine ....................................... .............. DTD: .................................. ...................... 0.50% Triethanolamine 98-99% .................... .............................. .DTD: ................. ......... 0.20% Water / preservatives ...... DTD: .......................... ............................................... DTD:. qs 100.00..DTD: - 11 -

  This final product can also be in the form of an emulsified gel which can be formulated as below: Polymer of acrylic acid ..........

  .................................................. ....... 0.60% Decamethylcyclopentasiloxane ...................................... ........ .DTD: ................... 3.00% Cetyl / Stearyl 2-Ethylhexanoate .......... .DTD: ............................................... .......... 7.00% Blackcurrant seed oil ............................... ........................ .DTD: .................... 0.77 % Palmitic acid ............................. DTD: ................ .............................................. 0.01% Vitamin B6 ................................................ ............... .DTD: ................................ 0.03% Serine .......................... DTD: ................ .................................................. ....... DTD: .... 0.06% Triethanolamine 98-99% ............................ ..... DTD: ........................................... 0.60% Water / preservatives .... DTD: .................................... ..................................... DTD: ... q.s. 100.00 The cosmetic composition comprising the amino-lipid vitamin complex according to the invention is a product for local application on the skin in order to maintain or increase its skin barrier by stimulating the synthesis of epidermal lipids. This product is particularly advantageous for treating dry, aged, damaged skin, because it reduces the loss of hydration, increases the suppleness of the stratum corneum and generally improves the quality of the skin. The use is simple and practical, a small amount of cream, or milk, or gel ..., is applied to the most exposed areas of the skin (face, neck, hands). The application is done either directly with the fingers or..DTD: using a spatula or cotton .... The clinical tests developed below

  after prove that notable results are obtained, after application, within 48 hours. A single application of the product may not be enough, it is necessary

  use this product continuously, for several consecutive days.

  Indeed, the final product (excipient + ALV complex) in the form of a cream has

  been tested by in vivo and in vitro studies.

  The in vitro measurement of the biosynthesis of epidermal lipids was obtained by bringing the skins into contact with a radiolabelled precursor of lipid biosynthesis (radiolabelled acetate) and then by evaluating the incorporation of radioactivity into the extracted epidermal lipids. after 24H or 48H

  application of the cream (ALV + vehicle complex).

  The evaluation of the effect on the metabolism of epidermal lipids of the amino-lipid vitamin complex with its excipient in the form of cream was carried out in parallel with that of its excipient taken in isolation by a study

  in vitro in a human skin explants model.

  A human skin sample was obtained from a tummy tuck performed on a woman aged 30. Explants of 8 mm in diameter were

  using a cookie cutter.

  The skin explants were cultivated in a basic culture medium, that is to say

  say the MEM medium supplemented with 10 mg / ml of glutamine, lg / l of glucose, 1 mg / ml of bovine serum albumin, 50 IU / ml of penicillin, 50 pg / ml of streptomycin and 0.2% of bicarbonate sodium but free of serine, palmitic acid,

  vitamin B6 and essential fatty acids.

  For each experimental condition, four explants were made.

  The test products (amino-lipid vitamin complex added to the vehicle or cream and vehicle only) were deposited pure on the surface of the explants, at a rate of 20 mg / cm 2, for 24 or 48 hours. The incubations were carried out at 37 ° C. in an atmosphere containing 5% of CO 2. Three pCi acetate labeled with carbon 14 (AMERSHAM, specific activity: 57 mCi / mmol) were added per ml of culture medium, 24 hours before the end of each incubation in order to monitor the production of lipids. At the end of each incubation, the explants of human skin were abundantly rinsed with PBS buffer. For three of the four explants from each experimental group, the epidermis was dissociated from the dermis by a controlled thermal shock (MilliQ water, 1 min, 60 ° C.). Each epidermis was then weighed. Conventionally, the fourth explant was fixed in 10% (v / v) formaldehyde for histological control of the explant morphology. It has been cut into several cutaways to find out if any

  morphological alterations existed.

  -13 - Time scales Incubation of 24 Hours To T24

I 0

A O

  48-hour incubation To T24 T48 lo 1 'A 1: topical application of the test products A: addition of acetate labeled with carbon 14 in the culture medium l: dermis / epidermis dissociation and evaluation of the effects O: histology neosynthesized total epidermal lipids, labeled with 14C carbon, were extracted by partition between an organic phase (methanol / chloroform, 1/4, v / v) and an aqueous phase (0.25 M potassium chloride). The organic phase was evaporated under nitrogen and the residues were taken up in a mixture

chloroform / methanol, 2/1 (v / v).

  The radioactivity of each sample, corresponding to the amount of acetate incorporated in the neosynthesized lipids and extracted, was measured in

  liquid scintillation. The results were expressed in cpm / mg of epidermis.

  The measurement of the incorporation of radiolabeled 14C carbon acetate in total epidermal lipids is completed by analysis by thin layer chromatography (TLC) to separate the main classes of lipids.

  neosynthesized radiolabelled epidermis.

  -14- From the extracted lipid samples, 20 μl aliquots, corresponding to approximately 40,000 cpm, were deposited on silica chromatography plates. These were developed in 3 successive solvents: - chloroform / acetone / methanol, 38/2/10 (v / v / v); chloroform / acetone / methanol, 40/5/5 (v / v / v); - chloroform / ethyl acetate / ether / methanol, 36/14/3/1 (vIvIvIvl); This system made it possible to separate the cholesterol sulfate, the cerebrosides, the ceramides, the cholesterol, the triglycerides plus the diglycerides. The polar lipids which remain at the deposition point mainly corresponded to

  phospholipids, they have been called "very polar lipids".

  The silica plates were then placed in opposition with films for

  autoradiography for 8 days (AMERSHAM, hyperfilm 13 max).

  The position of the different classes of lipids - very polar lipids, cholesterol sulfate, cerebrosides, ceramides, cholesterol, triglycerides plus diglycerides, a

  was determined using the corresponding standards.

  The radioactivity of the separate spots was counted with an argon-methane thin layer radioactivity analyzer (HERTHOLD). The results obtained corresponded to the percentages of the total radioactivity of each sample deposited on the TLC. Each of these percentages was multiplied by the total radioactivity obtained for each epidermis. (only is represented in

  the tables below the average of the results obtained with the three explants).

  To process the data, the data groups (control group and treated groups) were compared by a one-factor analysis of variance (ANOVA 1),

followed by a Donnett test.

  -15- Effect of the test products on the neosynthesis of total epidermal lipids in skin explants, after 24 and 48 hours of incubation

TIME TESTIMONIAL EXCIPIENT CREAM

6033 7980 * 6513

24 hours + 1- +/- +/-

1158 567 793

132 108

6537 13317 * 7170

48 hours +/- + 1- + + /

1747 3490 2019

204 110

  The results are expressed in cpm / mg of epidermis in bold: average of the results obtained in the three skin explants and corresponding standard deviation in italics: percentage of the control group *: mean significantly different from the control group (p <0.05) After 24 hours of incubation, a significant increase of a factor of 1.3 in the neosynthesis of total epidermal lipids was observed in the presence of the amino-lipid complex vitamin with its excipient (cream). This effect is not

  observed in the presence of the excipient alone.

  After 48 hours of incubation, the significant increase by a factor of 2 in the neosynthesis of total epidermal lipids was observed in the presence of the cream (amino-lipid vitamin complex + excipient). This effect was not

  observed in the presence of the excipient of the cream alone.

  In summary, under the experimental conditions adopted, the cream significantly increases the neosynthesis of total epidermal lipids in explants of human skin. This effect is greatest after 48 hours

incubation only after 24 hours.

  To determine the nature of the neosynthesized epidermal lipids, these were separated into large classes by thin layer chromatography. Very polar lipids, mainly phospholipids, and triglycerides +

  diglycerides represented the major peaks.

  -17 - Effect of the test products on the neosynthesis of polar deposition lipids, cholesterol sulphate, cerebrosides, ceramides, cholesterol and triqlyvcerides + diqlycerides. after 24 hours of incubation

LIPIDE TESTIMONIAL EXCIPIENT CREAM

Polar lipids 2195 3707 * 2551

of the deposit + 1- + 1- +/-

626,305,286

169 116

385,558,281

+ 1- + 1- +/- Sulfate

cholesterol 84 158 57

145 73

Cerebrosides 765 783 694

+ 1- + 1- + -'1

212 77 110

100 102 91

Ceramides 216 203 180

+ 1- + 1- +/-

9 35 39

94 83

Cholesterol 240 261 186

+/- + 1- +/-

28 37 35

109 77

Triglycerides 2232 3048 * 2279

+ + 1- + 1- +/-

Diglycerides 277 216 277

137 102

  The results are expressed in cpm / mg of epidermis in bold: average of the results obtained with the three skin explants and corresponding standard deviation in italics percentage of the control group *: mean significantly different from the control group (p <0.05)

- 18 -

  After 24 hours of incubation, the ALV complex with its excipient had no significant effect on the neosynthesis of the other epidermal lipids compared to the control explants cultivated in the basic medium. The cream increased by a factor of 1.7 the neosynthesis of very polar lipids, by a factor of 1.4 the neosynthesis of cholesterol sulfate and by a factor of 1.4 the neosynthesis of triglycerides and diglycerides. These effects were not observed in the presence of

the excipient for the cream (only).

- 19 -

  Effect of the test products on the neosynthesis of polar deposition lipids, cholesterol sulfate, cerebrosides, ceramides, cholesterol and triglycerides + diqlycerides, after 48 hours of incubation

LIPIDE TESTIMONIAL EXCIPIENT CREAM

Polar lipids 2214 4488 * 2484

of the deposit +/- + 1- +/-

932 1,799,856

203 11

304 1060 * 338

+ 1- +/- +/- sulfate

cholesterol 52 488 186

348 11

Cerebrosides 430 366 618

+ 1- + 1- +/-

108,301,200

85,144

Ceramides 115 117 69

+/- + 1- +/-

72 54 33

101 60

Cholesterol 146 389 * 207

+ 1- + 1- +/-

43 127 55

266,142

Triglycerides 3228 7987 * 3556

+ + 1- + 1- +/-

Diglycerides 871 2866 844

247,110

  The results are expressed in cpm / mg of epidermis in bold: average of the results obtained with the three skin explants

and corresponding standard deviation.

  in italics percentage of the control group *: mean significantly different from the control group (p <0.05)

- 20 -

  After 48 hours of incubation, the cream increased by a factor of 2 the neosynthesis of very polar lipids, by a factor of 3.5 the neosynthesis of cholesterol sulfate, by a factor of 2.7 that of cholesterol and a factor 2.5 that of triglycerides + diglycerides. These effects were not observed in the presence of the excipient of the cream. Effect of the test products on the neosynthesis of cerebrosides. of cholesterol sulfate of cholesterol and all the lipids of cohesion, after 24 hours of incubation

LIPIDE TESTIMONIAL EXCIPIENT CREAM

Cerebrosides 981 986 874

+ +/- + 1- +/-

Ceramides 213 42 117

101 89

Cholesterol 625 819 467

+ + 1- + 1- +/-

  Sulphate 97 194 87 cholesterol 100 131 75 Total lipids 1606 1805 1341

previous +/- + 1- +/-

= 306 219 203

  Lipids of the 100 112 83 cohesion The results are expressed in cpm / mg of epidermis in average fat of the results obtained with the three skin explants and corresponding standard deviations in italics percentage of the control group -21 - Effect of the test products on neosynthesis of cerebrosides. cholesterol + cholesterol sulfate and all cohesion lipids, after 48 hours of incubation

LIPIDE TESTIMONIAL EXCIPIENT CREAM

Cerebrosides 546 482 689

+ + 1-. + 1- + 1-

Ceramides 158,321,217

88,126

Cholesterol 450 1449 * 545

+ + 1- +/- +/-

  Sulfate 78,572,230 cholesterol 100,322,121 Total lipids 996 1,932 * 1,233

previous + 1- +/- +/- +1 -

= 220,433,416

  Lipids of 100 194 124 cohesion The results are expressed in cpm / mg of epidermis in bold average of the results obtained with the three skin explants and corresponding standard deviations in italics percentage of the control group *: mean significantly different from the control group (p < 0.05) The cream increases by 35% at 24 hours and by 60% at 48 hours also the neosynthesis of minor lipids quantitatively but essential from a functional point of view, those of cellular cohesion. This increase concerns cholesterol and cholesterol sulfate after 48 hours of application as shown by the results of

tables above.

  -22 - In conclusion, the increase in neosynthesis of total epidermal lipids 22% at 24 hours and 85% at 48 hours caused by the cream results mainly from an increase in the neosynthesis of phospholipids (polar lipids of the deposit), triglycerides and diglycerides, cholesterol and cholesterol sulfate. These effects were greatest after 48 hours of incubation

  only after 24 hours. The excipient for the cream had no effect.

  By applying said cream, the quality of the skin is thus greatly improved in its lipid portion. We see that the neosynthesis of these lipids

epidermal is favored.

  The skin does not get used to this cream (complex + excipient) and remains very active, ready to synthesize again everything it needs, and this, in the extreme case where it is cut off from its natural sources of supply.

  as shown by this experience on female skin taken out of context.

  Everything happens as if, after a treatment with this cream, the synthesis of epidermal lipids is improved to the great benefit of dry, sensitive or atopic skin. In addition, oily skin can also use it, because the increase in said lipid synthesis does not concern the lipids synthesized by the sebaceous glands but only the lipids synthesized by keratinocytes (epidermal cells). The neosynthesis of the cholesterol / Cholesterol sulfate by the said cream also allows the prevention of cutaneous carcinomas. Said cream can be used as a preventive measure and also when accidentally the person has been exposed to a carcinogenic promoter. An in vivo study on a panel of 30 women was also carried out. The latter used this cream against its placebo in combination with a gel containing two trace elements (Mg and Zn) as well as a compound inhibiting 5 a reductase, the enzyme responsible for the cutaneous production of dihydrosterone,

highly androgenic molecule.

  -23 - The results of this study using analog scales to qualify the effect of this treatment combining the cream containing the amino-lipid vitamin complex and a gel inhibiting androgenic skin activity show: () a decrease in expression lines in favor of the treated group (GT) compared to the placebo group (GP) c> 35% for GT and 5.18% for GP;

  (O) an improvement in the general quality of the skin> 8.82% for GT and -

  9.10% for GP; () resistance of the skin to external aggressions 34.43% for GT and 12.14% for GP; smoother skin * 29.78% for GT and - 13.49% for GP; (softer skin r 24.77% for GT and -7.29% for GP; () a brighter complexion> 70% for GT and 16.86% for GP; a reduction in the down of the facial skin> 23.93% for GT and 11.26% for GP; (a decrease in fatigue marks ó 54.26% for GT and 2.66% for GP. All of these results show that the effects highlighted ex vivo are confirmed in vivo, and this particularly for the parameters (wrinkles, general quality, resistance to attack, luminosity of the complexion and can be reduced

tiredness).

  This effect is amplified by the inhibition of androgenic activity which provides a

  smoother, softer skin, with less unsightly down.

-24-

Claims (8)

  1. Vitamin amino-lipid complex constituted in particular by precursors of the biosynthesis of ceramides, characterized in that it comprises serine or a precursor of serine, vitamin B6,   vegetable oil and palmitic acid.   2. Vitamin amino-lipid complex constituted in particular by precursors of the biosynthesis of ceramides according to claim 1   characterized in that the vegetable oil is black currant seed oil.   3. Vitamin amino-lipid complex constituted in particular by precursors of the biosynthesis of ceramides according to claim 1 or 2 characterized in that palmitic acid is between 0, 5 and 40% in that the serine or its precursor is a concentration of 3 to 30%, in that the vegetable oil is between 40 and 80% and in that the vitamin B6 is between 3 and 10%.   4. Vitamin amino-lipid complex constituted in particular by precursors of the biosynthesis of ceramides according to claim 3 characterized in that the respective concentrations of the elements of the amino-lipid vitamin complex are: palmitic acid: 1%, serine: 16   %, vegetable oil: 77%, vitamin B6: 6%.   5. Cosmetic composition characterized in that it comprises the complex   Vitamin amino lipid according to any one of Claims 1 to 4 a   a rate between 0.5 and 10% and preferably between 1 and 5%.   6. Cosmetic composition according to claim 5 characterized in that it   contains the amino-lipid vitamin complex at a rate of 3%.   7. Cosmetic composition according to Claim 5, characterized in that it is a water-in-oil emulsion in the form of a cream, the formula of which is preferably the following: Decamethylcyclopentasiloxane / Dimethylsiloxane glycol copolymer. ......... 18.00% Dimethyl silicone / Trimethylsiloxysilicate .................   ................................ 5.00% Decamethylcyclopentasiloxane / aluminum magnesium stearate ......... ....... CLMF: ... 1.50% Dimethyl silicone ................................ ............. CLMF: ................................... ....... 3.60% Palmitic acid ....... CLMF: ............................ ............................................. CLMF: ... ......... 0.40% Blackcurrant seed oil ............................ CLMF: .. ............................................ 2.00% Vitamin B6 ... CLMF: ............................................. ............................ CLMF: .................... .... 0.03% Serine .................................... CLMF: ... .................................................. ................. 0.03% Sodium chloride .......................... ....................... .CLMF: ........................ .............. 1.00% Water / preservatives ........... CLMF: ................ .................................................. .. q. s. 100.00 8. Cosmetic composition according to Claim 5, characterized in that it is an oil-in-water emulsion in the form of a cream, the formula of which is preferably the following: Sorbitan monostearate ... ............................................... .CLMF: ....................... 3.26% Cetyl phosphate + cethanol phosphate diethanolamine ............... .............. 2.72% Isocetyl stearate ............................ ................................... .CLMF: ............ ............ 4.00% Blackcurrant seed oil ................. CLMF: .......... .................................................. 0.80% Palmitic acid ............................................ .............. .CLMF: .............................. 0.20 % Dicaprylate / Propylene glycol dicaprate ........................................... ....... 4.00% Polyethylene glycol 1500 .................................... .............. .CLMF: ............ 1.00% Triglycerides of gr acids as saturated ............. CLMF: ................................. .............. 4.00% Copolymer of acrylic acid .......................... ............................... .CLMF: .... 0.25% Vitamin B6 ...... ............................................ CLMF: .... ............................................. 0.06% Triethanolamine 98-99% .............................................. ..... .CLMF: .................... 0.25% Serine ................. ...................... CLMF: .......................... ....................................... 0.16% Water / preservatives .... ............................................ CLMF: .... .......................... qs 100.00 -26- 9. Cosmetic composition according to claim 7 or 8 characterized in that a 5 alpha reductase inhibitor is added at a rate between 0, 5 and 5%, and preferably 3% 10. Composition cosmetic according to Claim 5, characterized in that it is a water-in-oil emulsion in the form of a milk, the formula of which is preferably the following: Sorbitan mono-oleate / ethoxylated hydrogenated castor oil / hydrogenated castor oil / beeswax ................... CLMF: ................... ............................... 4.00% hydrogenated castor oil POE (7) ...................................... .CLMF: ...... ............... 2.00% fluid vaseline oil .................... CLMF: ..... .................................................. ..... 10.00% Isopropyl palmitate ..................................... ............... .CLMF: ....................... 5.00% Triglycerides of capric acid / caprylic ................................................ 10.00% Blackcurrant seed oil ........................................ .CLMF: .................................... 0.77% Palmitic acid .... ..... CLMF: ........................................... .............................. CLMF: ........ 0.01% Vitamin B6 .... .......................................... CLMF: ...... ............................................... 0.03 % Serine ...... CLMF: ........................................ ................................. CLMF: ............... ............. 0.03% Magnesium sulfate 7H20 ... CLMF: ........................ ......................... ............... 0.70% Water / preservatives ............................ ........................... .CLMF: ................... qs 100.00 -27- 11. Cosmetic composition according to Claim 5, characterized in that it is an oil-in-water emulsion in the form of a milk, the formulation of which is preferably the following: Monostearate sorbitan ..................................... .CLMF: ......... ............................... 2.82% Isopropyl Palm ............ ..................................... CLMF: ........... ................ 9.00% thick petroleum jelly ........................... ........................................ .CLMF: ....... ... 1.00% Cetyl alcohol ............... CLMF: ........................ ................................................. CLMF : 0.50% Cetyl ether P.OE (20) ................. CLMF: ............................ ............................... 2.18% Methyl phenyl polysiloxane ............ ................ CLMF: ................................ ............. 0.50% Blackcurrant seed oil ...................... CLMF: ..... ................................................ 3, 00% Palmitic acid .............................................. ....... CLMF: ....................................... 1, 00% Acrylic acid polymer ........................................... CLMF: ......................... 0.18% Vitamin B6 ........... CLMF: ... .................................................. .................... CLMF: ........... 0.30% Serine ............ ....................... CLMF: ......................... ................................................ CLMF: 0.50% Triethanolamine 98-99% ................. CLMF: ...................... ............................... 0.18% Water / preservatives ............ ........................................... ... CLMF: ........................ q.s. 100.00 12. Cosmetic composition according to Claim 5, characterized in that it is a liposome cream, the formulation of which is preferably the following: Cetyl alcohol polyglycerol ............. ................................. .CLMF: ............ 3, 80% B-sitosterol ...................................... CLMF: ..... .................................................. .. 3.80% Diketyl phosphate .......................................... ...... CLMF: .................................... 0.40% Sunflower oil .................................................. ............... .CLMF: .................... 33.00% Blackcurrant seed oil .... .................................................. .... .CLMF: ................ 2.00% Palmitic acid ........ CLMF: ........... .................................................. ............ CLMF: ....... 0.05% Polymer of acrylic acid ................ CLMF: .. .................................................. 0.20% Vitamin B6 ... .................................................. .......... .CLMF: .................................. 0.30 % Serine ................................................ .................... .CLMF: ........................... .. 0.50% Triethanolamine 98- 99% ....................................... ............................... .CLMF: ...... 0.20% Water / preservatives ... ..................... CLMF: ........................... ............................. qs 100.00 - 2813. Cosmetic composition according to Claim 5, characterized in that it is a gel the formulation of which is preferably the following: Polymer of acrylic acid .......... ......................... .CLMF: ...................... ........... 0.60% Decamethylcyclopentasiloxane .... CLMF: ............................ .................................. 3.00% Cetyl / Stearyl 2-Ethylhexanoate ...... ................................... CLMF: ............. ................ 7.00% Blackcurrant seed oil ............ CLMF: ........... .................................................. ..... 0.77% Palmitic acid ....................................... ................. .CLMF: .............................. ... 0.01% Vitamin B6 ..................... CLMF: .................. .................................................. ..... CLMF: .. 0.03% Serine .................................... ..................... CLMF: ........................... ..................... 0.06% Sorted thanolamine 98-99% ............................................. ...... .CLMF: ........................ 0.60% Water / preservatives .......... .............. CLMF: .................................. ........................ qs 100.00 14. Process for the synthesis of epidermal lipids in vivo, characterized in that it consists in applying to the skin, a cosmetic composition according to one..CLMF: any of claims 5 to 13 or else the amino-lipid complex   Vitamin after shaking according to any one of claims 1 to 4.   15. Cosmetic composition according to any one of claims 5 to 13,   and vitamin amino-lipid complex according to any one of   Claims 1 to 4, characterized in that they are used as a preventive measure skin cancer.   16. Cosmetic process characterized in that the cosmetic composition according to any one of the following is applied to the skin simultaneously   claims 5 to 8 and 10 to 13, or else the ALV complex after stirring   according to any one of claims 1 to 4, and a gel containing at   minus a 5 alpha reductase inhibitor.