Dermatological Cosmetic Composition Comprising an SDKP Peptide or an Analog Thereof

Abstract

The invention relates to a lipid nanoparticle comprising an SDKP peptide conjugate or a biological peptide analog thereof, a method for obtaining said lipid nanoparticle, and the cosmetic use thereof as an anti-wrinkle agent or skin restructuring agent.

Claims

1. A peptide comprising a peptide strand of formula SDKP, or a biological peptide analog thereof, wherein at least one linear or branched, saturated or unsaturated, C.sub.13-C.sub.50 fatty chain is grafted at the N- and/or C-terminal end of the peptide strand.

2. The peptide conjugate as claimed in claim 1, wherein the biological analog comprises a peptide strand of formula (II) below:
—Y.sub.1—Y.sub.2—Y.sub.3—Y.sub.4—  (II) wherein, Y.sub.1 represents the N-terminal end of the peptide strand (II), the Y.sub.3—Y.sub.4 or —Y.sub.4 fragments possibly being absent from the peptide strand of formula (II), Y.sub.1 represents serine or a structural analog of serine, Y.sub.2 represents aspartic acid or a structural analog of aspartic acid, Y.sub.3 represents lysine or a structural analog of lysine, and Y.sub.4 represents proline or a structural analog of proline.

3. The peptide conjugate or biological analog thereof as claimed in claim 1, wherein said conjugate is of formula (I) below:
R.sub.1—X.sub.1—X.sub.2—X.sub.3—X.sub.4—A.sub.1—R.sub.2   (I) wherein: R.sub.1, which is on the N-terminal side, is a hydrogen atom, or a fatty chain chosen from a substituted or unsubstituted, linear or branched C.sub.13-C.sub.50 alkyl group, a substituted or unsubstituted, linear or branched C.sub.21-C.sub.50 alkylaryl group, a substituted or unsubstituted, linear or branched C.sub.13-C.sub.50 alkenyl group, or an R.sub.3—CO—, R.sub.3—OCO— or R.sub.3—COO— group wherein R.sub.3 is a substituted or unsubstituted, linear or branched C.sub.13-C.sub.49 alkyl or alkenyl group, the substitutions including F, Cl, or any lipophilic heteroatom or heteroatom group; X.sub.1 is an (L)-serine condensate, X.sub.2 is an (L)- and/or (D)-aspartic acid or (L)- and/or (D)-glutamic acid condensate, X.sub.3 is an (L)- and/or (D)-lysine, (L)- and/or (D)-arginine or (L)- and/or (D)-ornithine condensate, X.sub.4 is an (L)- and/or (D)-proline condensate, X.sub.4, or X.sub.3 and X.sub.4, optionally being absent, the bonds linking X.sub.1 to X.sub.2, X.sub.2 to X.sub.3 where appropriate, and X.sub.3 to X.sub.4 where appropriate, possibly being peptide or pseudopeptide bonds, A.sub.1 is a covalent bond, an NH group or an oxygen atom, R.sub.2 is a hydrogen atom, or a fatty chain chosen from a substituted or unsubstituted, linear or branched C.sub.13-C.sub.50 alkyl group, a substituted or unsubstituted, linear or branched C.sub.21-C.sub.50 alkylaryl group, a substituted or unsubstituted, linear or branched C.sub.13-C.sub.50 alkenyl group, or an R.sub.4—CO— or R.sub.4—OCO— group wherein R.sub.4 is a substituted or unsubstituted, linear or branched C.sub.13-C.sub.50 alkyl or alkenyl group, the substitutions including F, Cl, or any lipophilic heteroatom or heteroatom group, R.sub.1 and R.sub.2 are not hydrogen atoms, or a pharmaceutically acceptable salt thereof.

4. A lipid nanoparticle comprising: a core consisting of a lipid phase (L.sub.1); at least one surfactant comprising a hydrophilic portion and a lipophilic portion; an internal membrane surrounding said core consisting of a lipid phase (L.sub.2), comprising the lipophilic portion of said surfactant; an external membrane, surrounding said internal membrane, consisting of an aqueous phase (A.sub.1) comprising the hydrophilic portion of said surfactant; and at least one peptide conjugate, comprising a peptide strand of formula SDKP, or a biological peptide analog thereof, wherein at least one linear or branched, saturated or unsaturated C.sub.3-C.sub.50 fatty chain is grafted at the N- and/or C-terminal end of the peptide strand, and said peptide conjugate is such that its lipophilic portion is in the lipid phase L.sub.2 and its hydrophilic portion is in the phase A.sub.1.

5. The lipid nanoparticle as claimed in claim 4, wherein the peptide conjugate or the biological analog thereof is a peptide conjugate comprising a peptide strand of formula SDKP, or a biological peptide analog thereof, wherein at least one linear or branched, saturated or unsaturated C.sub.13-C.sub.50 fatty chain is grafted at the N- and/or C-terminal end of the peptide strand.

6. The lipid nanoparticle as claimed in claim 4 further comprising the peptide conjugate of formula Ac-SDKP-OH, Ac-SDKP-NH.sub.2 or a mixture thereof.

7. The lipid nanoparticle as claimed claim 4, wherein the peptide conjugate is of formula (I):
R.sub.1—X.sub.1—X.sub.2—X.sub.3—X.sub.4—A.sub.1—R.sub.2   (I) wherein: R.sub.1, which is on the N-terminal side, is a hydrogen atom, a substituted or unsubstituted, linear or branched C.sub.3-C.sub.50 alkyl group, a substituted or unsubstituted, linear or branched C.sub.8-C.sub.50 alkylaryl group, a substituted or unsubstituted, linear or branched C.sub.3-C.sub.50 alkenyl group, or an R.sub.3—CO—, R.sub.3—OCO— or R.sub.3—COO— group wherein R.sub.3 is a substituted or unsubstituted, linear or branched C.sub.3-C.sub.50 alkyl or alkenyl group, the substitutions including F, Cl, or any lipophilic heteroatom or heteroatom group, and X.sub.1 is an (L)-serine condensate, X.sub.2 is an (L)- and/or (D)-aspartic acid or (L)- and/or (D)-glutamic acid condensate, X.sub.3 is an (L)- and/or (D)-lysine, (L)- and/or (D)-arginine or (L)- and/or (D)-ornithine condensate, X.sub.4 is an (L)- and/or (D)-proline condensate, X.sub.4, or X.sub.3 and X.sub.4 optionally being absent, the bonds linking X.sub.1 to X.sub.2, X.sub.2 to X.sub.3 where appropriate, and X.sub.3 to X.sub.4 where appropriate, being peptide or pseudopeptide bonds, A.sub.1 is a covalent bond, an NH group or an oxygen atom, R.sub.2 is a hydrogen atom, a substituted or unsubstituted, linear or branched C.sub.3-C.sub.50 alkyl group, a substituted or unsubstituted, linear or branched C.sub.8-C.sub.50 alkylaryl group, a substituted or unsubstituted, linear or branched C.sub.3—C.sub.50 alkenyl group, or an R.sub.4-CO— or R.sub.4—OCO— group wherein R.sub.4 is a substituted or unsubstituted, linear or branched C.sub.3-C.sub.50 alkyl or alkenyl group, the substitutions including F, Cl, or any lipophilic heteroatom or heteroatom group R.sub.1 and R.sub.2 not being able to both be hydrogen atoms, or a pharmaceutically acceptable salt thereof.

8. The lipid nanoparticle as claimed in claim 4, wherein: R.sub.1 is a substituted or unsubstituted, linear or branched C.sub.6-C.sub.30 alkyl group or an R.sub.3—CO—group, X.sub.1 is an (L)-serine condensate, X.sub.2 is an (L)- and/or (D)-aspartic acid or (L)- and/or (D)-glutamic acid condensate, X.sub.3 is an (L)- and/or (D)-lysine, (L)- and/or (D)-arginine or (L)- and/or (D)-ornithine condensate, X.sub.4 is an (L)- and/or (D)-proline condensate, X.sub.4 or X.sub.3 and X.sub.4, optionally being absent, the bonds linking X.sub.1 to X.sub.2, X.sub.2 to X.sub.3 where appropriate, and X.sub.3 to X.sub.4 where appropriate, possibly being peptide or pseudopeptide bonds, A.sub.1 is a covalent bond, an NH group or an oxygen atom, R.sub.2 is a substituted or unsubstituted, linear or branched C.sub.5-C.sub.30alkyl or alkenyl group, the substitutions including F, Cl, or any lipophilic heteroatom or heteroatom group such as a guanidine or guanidinium group or a pharmaceutically acceptable salt thereof.

9. The lipid nanoparticle as claimed in claim 4, wherein said lipid nanoparticle comprises at least one of the following constituents: at least one C.sub.5-C.sub.30 fatty acid, which is optionally hydrogenated and/or in glycol ester form, in the (L.sub.1) phase, a C.sub.1-C.sub.30 fatty acid ester of polyoxyethylene (10-100) as surfactant, at least one C.sub.5-C.sub.30 fatty acid, which is optionally hydrogenated and/or in glycol ester form, in the (L.sub.2) phase, and/or at least one preservative in the (Ai) phase.

10. The lipid nanoparticle as claimed in claim 4, wherein the peptide conjugate comprises the sequence (L)S-(L)D-(L)K-(L)P.

11. A method for producing a lipid nanoparticle as claimed in claim 4, wherein said method comprises the following steps: a. preparing a lipid phase and an aqueous phase, at least one of the two phases comprising a surfactant, at least one of the two phases comprising the peptide strand of formula SDKP, or a biological peptide analog thereof, grafted with a fatty chain chosen from a substituted or unsubstituted, linear or branched C.sub.13-C.sub.50 alkyl group, a substituted or unsubstituted, linear or branched C21-C.sub.50 alkylaryl group, a substituted or unsubstituted, linear or branched C13-C.sub.50 alkenyl group, or an R.sub.3—CO—, R.sub.3—OCO— or R.sub.3—COO—group wherein R.sub.3 is a substituted or unsubstituted, linear or branched C.sub.13-C.sub.49 alkyl or alkenyl group or an R.sub.4—CO—or R.sub.4—OCO— group wherein R.sub.4 is a substituted or unsubstituted, linear or branched C13-C.sub.50 alkyl or alkenyl group, the substitutions including F, Cl, or any lipophilic heteroatom or heteroatom group; b. emulsifying said lipid phase and said aqueous phase, resulting in the formation of lipid nanoparticles, optionally under pressure; c. recovering the lipid nanoparticles formed.

12. A cosmetic formulation comprising one or more lipid nanoparticles as claimed in claim 4.

13. The cosmetic formulation as claimed in claim 12, comprising between 1% and 25% by weight of nanoparticles comprising: a core consisting of a lipid phase (L.sub.1); at least one surfactant comprising a hydrophilic portion and a lipophilic portion; an internal membrane surrounding said core consisting of a lipid phase (L.sub.2), comprising the lipophilic portion of said surfactant; an external membrane, surrounding said internal membrane, consisting of an aqueous phase (A.sub.1) comprising the hydrophilic portion of said surfactant; and at least one peptide conjugate, comprising a peptide strand of formula SDKP, or a biological peptide analog thereof, wherein at least one linear or branched, saturated or unsaturated C.sub.3-C.sub.50 fatty chain is grafted at the N- and/or C-terminal end of the peptide strand, and said peptide conjugate is such that its lipophilic portion is in the lipid phase L2 and its hydrophilic portion is in the phase Ai.

14. A nanoemulsion comprising: at least one dispersed lipid phase (L.sub.3), and at least one continuous aqueous phase (A.sub.2) wherein the aqueous phase comprises at least one surfactant, wherein, at the interface between the aqueous phase (A.sub.2) and the lipid phase (L.sub.1), there is at least one peptide conjugate comprising a peptide strand of formula SDKP, or a biological peptide analog thereof, wherein at least one linear or branched, saturated or unsaturated, C.sub.13-C.sub.50 fatty chain is grafted at the N- and/or C-terminal end of the peptide strand and/or in that said nanoemulsion comprises at least one lipid nanoparticle comprising a core consisting of a lipid phase (L.sub.1); at least one surfactant comprising a hydrophilic portion and a lipophilic portion; an internal membrane surrounding said core consisting of a lipid phase (L.sub.2), comprising the lipophilic portion of said surfactant; an external membrane, surrounding said internal membrane, consisting of an aqueous phase (A.sub.1) comprising the hydrophilic portion of said surfactant; and at least one peptide conjugate, comprising a peptide strand of formula SDKP, or a biological peptide analog thereof, wherein at least one linear or branched, saturated or unsaturated C.sub.3-C.sub.50 fatty chain is grafted at the N- and/or C-terminal end of the peptide strand, and said peptide conjugate is such that its lipophilic portion is in the lipid phase L.sub.2 and its hydrophilic portion is in the phase A.sub.1 wherein optionally the lipids of (L.sub.1) and (L.sub.3), or (L.sub.2) and (L.sub.3) are identical.

15. A method for restricting or preserving the appearance of the skin of a human subject comprising a step of applying on the skin a peptide conjugate comprising a peptide strand of formula SDKP, or a biological peptide analog thereof, wherein at least one linear or branched, saturated or unsaturated, C.sub.13-C.sub.50 fatty chain is grafted at the N- and/or C-terminal end of the peptide strand.

16. The method as claimed in claim 15 wherein restricting or preserving the appearance of the skin includes anti-aging the skin.

17. A method for restructuring or preserving the appearance of the skin of a human subject comprising a step of applying on the skin a lipid nanoparticle comprising: a core consisting of a lipid phase (L.sub.1); at least one surfactant comprising a hydrophilic portion and a lipophilic portion; an internal membrane surrounding said core consisting of a lipid phase (L.sub.2), comprising the lipophilic portion of said surfactant; an external membrane, surrounding said internal membrane, consisting of an aqueous phase (A.sub.1) comprising the hydrophilic portion of said surfactant; and at least one peptide conjugate, comprising a peptide strand of formula SDKP, or a biological peptide analog thereof, wherein at least one linear or branched, saturated or unsaturated C.sub.3-C.sub.50 fatty chain is grafted at the N- and/or C-terminal end of the peptide strand, and said peptide conjugate is such that its lipophilic portion is in the lipid phase L.sub.2 and its hydrophilic portion is in the phase A.sub.1.

18. A method for restructuring or preserving the appearance of the skin of a human subject comprising a step of applying on the skin a nanoemulsion comprising: at least one dispersed lipid phase (L.sub.3), and at least one continuous aqueous phase (A.sub.2) wherein the aqueous phase comprises at least one surfactant, wherein, at the interface between the aqueous phase (A.sub.2) and the lipid phase (L.sub.1), there is at least one peptide conjugate comprising a peptide strand of formula SDKP, or a biological peptide analog thereof, wherein at least one linear or branched, saturated or unsaturated, C.sub.13-C.sub.50 fatty chain is grafted at the N- and/or C-terminal end of the peptide strand and/or in that said nanoemulsion comprises at least one lipid nanoparticle comprising: a core consisting of a lipid phase (Li); at least one surfactant comprising a hydrophilic portion and a lipophilic portion; an internal membrane surrounding said core consisting of a lipid phase (L.sub.2), comprising the lipophilic portion of said surfactant; an external membrane, surrounding said internal membrane, consisting of an aqueous phase (A.sub.1) comprising the hydrophilic portion of said surfactant; and at least one peptide conjugate, comprising a peptide strand of formula SDKP, or a biological peptide analog thereof, wherein at least one linear or branched, saturated or unsaturated C3-C.sub.50 fatty chain is grafted at the N-and/or C-terminal end of the peptide strand, and said peptide conjugate is such that its lipophilic portion is in the lipid phase L.sub.2 and its hydrophilic portion is in the phase A.sub.1. wherein optionally the lipids of (L.sub.1) and (L.sub.3), or (L.sub.2) and (L.sub.3) are identical.

19. A method for restructuring or preserving the appearance of the skin of a human subject comprising a step of applying on the skin a cosmetic formulation comprising one or more lipid nanoparticles comprising: a core consisting of a lipid phase (L.sub.1); at least one surfactant comprising a hydrophilic portion and a lipophilic portion; an internal membrane surrounding said core consisting of a lipid phase (L.sub.2), comprising the lipophilic portion of said surfactant; an external membrane, surrounding said internal membrane, consisting of an aqueous phase (A.sub.1) comprising the hydrophilic portion of said surfactant; and at least one peptide conjugate comprising a peptide strand of formula SDKP, or a biological peptide analog thereof, wherein at least one linear or branched, saturated or unsaturated C.sub.3-C.sub.50 fatty chain is grafted at the N- and/or C-terminal end of the peptide strand, and said peptide conjugate is such that its lipophilic portion is in the lipid phase L.sub.2 and its hydrophilic portion is in the phase A.sub.1.

Description

FIGURES

[0146] The histogram of FIG. 1 represents the variations in the isotropy following the comparative study according to example 4 below. The product B comprises lauroyl-SDKP-OH, whereas the placebo A is a neogoutte formulation free of SDKP analogs. The significant effect on the relief of the skin (isotropy—orientation of the lines of the skin) of the product B compared with the placebo can be seen.

[0147] The histogram of FIG. 2 represents the variations in the density of the dermis following the comparative study according to example 4 below. The product B comprises lauroyl-SDKP-OH, whereas the placebo A is a neogoutte formulation free of SDKP analogs. The significant effect on the density of the dermis of the product B compared with the placebo can be seen.

[0148] The histogram of FIG. 3 represents the overall assessment and organoleptic characteristics of the products (ease of application, light and pleasant texture, rapid penetration, application, non-greasy product, fragrance). It can be seen that the formulations tested have organoleptic characteristics and general assessments which are similar and acceptable.

EXAMPLES

Example 1: Synthesis of the “lauroyl-SDKP-OH” Compound

[0149] The synthesis described below is for illustration purposes. It can for example be carried out on a solid support (resin) by Fmoc strategy (on an automated device for amounts less than 50 mg and manually for larger amounts). The levels of charges of the resins and the concentrations of the reagents (coupling agents, bases, acids, scavengers) can vary according to the common practice in the art.

[0150] The starting materials used can be: [0151] natural amino acids protected on their N-terminal ends (Fmoc protection) and on their side chains (acido-labile protective groups which are therefore compatible with the Fmoc strategy); [0152] activators (HOBT/DIC). These products are eliminated by washing throughout the synthesis, then during the precipitation phase and, finally, during the purification phase; [0153] washing solvents (DMF, DCM). These products are eliminated by washing throughout the synthesis, then during the precipitation phase and, finally, during the purification phase; [0154] Wang resin precharged with the Fmoc-L-Pro amino acid. The initial, or reduced, level of proline charge of the resin can vary, but is typically between 0.1 and 2.0 mol/g of resin, preferentially between 0.5 and 1.5 mol/g, more preferentially approximately 0.75 mol/g ±10%. The resin is eliminated by simple filtration at the end of the synthesis; [0155] the usual bases for removing Fmoc are used, such as triethylamine, DBU, piperidine, etc. Piperidine was in particular used here; [0156] the usual acids for removing the peptide from the support were used (e.g. TFA, HCI, acetic acid, etc.). In the case of “Wang” resins, TFA (trifluoroacetic acid) with a scavenger, such as usual silicon derivatives in the art (e.g. TIS).

[0157] The synthesis is carried out in the following way: [0158] Starting point: Resin precharged with the Fmoc-Proline; [0159] 1. Grafting of the second amino acid, lysine: [0160] Deprotection of the supported proline with piperidine (for example 20% in DMF) in order to remove the Fmoc group; [0161] 3 series of washes with DMF; [0162] Coupling of Fmoc-L-Lys(Boc)-OH on the extending peptide, by using HOBt, DCI; [0163] 3 series of washes with DMF; [0164] Acetylation using acetic anhydride or the equivalent acyl chloride in order to block any unreacted amines of Pro; [0165] 3 series of washing with DMF; [0166] 2. Grafting of the aspartic acid Fmoc-L-Asp(OtBu)-OH: [0167] The same steps as for the grafting of lysine are reproduced for aspartic acid. [0168] 3. Grafting of the serine Fmoc-L-Ser(tbu)-OH: [0169] The same steps as for the grafting of lysine/aspartic acid are reproduced for serine. [0170] 4. Addition of the lauroyl fragment on the amine of the serine [0171] Deprotection of the serine with piperidine (20% in DMF) in order to remove the Fmoc group; [0172] 3 series of washes with DMF; [0173] addition of lauroyl chloride in the presence of pyridine, addition followed by ninhydrin test (conventional in the art) until complete conversion; [0174] 3 series of washes with DMF; [0175] 5. Cleavage of the peptide from the resin and deprotection [0176] treatment of the resin with a 95/2.5/2.5 TFA/TIS/H.sub.2O mixture [0177] 3 precipitations with ether, centrifugation. [0178] Solution is made with a part of the pellet for analysis.

[0179] The post-synthesis analysis is carried out as follows: [0180] The analysis is carried out on a Waters Acquity H Class UPLC and on a 150×2.1 mm BEH C18 column: [0181] 0 to 100% of acetonitrile in 8.5 min, then 100% to 0% of acetonitrile in 2 min, flow rate: 0.6 ml/min.

[0182] The mass spectrometry analysis is carried out on a Waters LC/MS system composed of 2695 separation module alliance; X-Bridge C18 column, 3.5 p.m, 4.6 x 150 mm; 2996 photodiode array detector; SQ mass detector 2 (analyzer: quadripole and source : ES+), flow rate: 0.6 ml/min: [0183] 3 to 97% of acetonitrile in 15 min, [0184] then a plateau at 97% of acetonitrile for 2 min, [0185] then 97% to 3% of acetonitrile in 2 min, then a plateau at 3% of acetonitrile for 2 min.

[0186] The purification is carried out as follows: [0187] The purification is carried out on a Waters LC4000 preparative HPLC and on a Vydac denali column 50×300 mm, 10 micron, in an H.sub.2O (0.1% TFA)/acetonitrile (0.1% TFA) mixture.

[0188] The post-purification analysis is carried out as follows: [0189] Analysis of the fractions collected on HPLC according to the protocol described above. The fractions corresponding to the specifications are combined, freeze-dried, and then re-analyzed by HPLC and by mass spectrometry according to the protocols described above. [0190] The final product essentially consists of the lauroyl-SDKP-OH peptide, at more than 90%. [0191] The impurities present are deletion peptides such as Ac-P-OH, Ac-KP-OH or Ac-DKP-OH. [0192] The solvents, activators, protections, . . . are eliminated as the synthesis proceeds, during the precipitation with ether (volatile) and then during the purification and, finally, the freeze-drying.

Example 2: Neogoutte Preparation

[0193] A neogoutte according to the present invention can be prepared according to the procedure disclosed in FR 2 991 196 on the basis of the amounts according to Table 1 below:

TABLE-US-00001 TABLE 1 Compounds involved in the preparation of the neogoutte: Weight % Trade Weight in the Compound name Supplier (mg) method Aqueous Water — — 1500 75.00 phase PEG 40 Myrj s40 Croda 215 10.75 stearate Lipid Phospho- Phospho- Lipoid 45 2.25 phase lipids lipon Olive oil 115 5.75 Wax Lipocire Gattefossé 115 5.75 Palmitoyl- — Creative 9.99 0.4995 KTTKS Peptide 1% solution Gen Pep 0.01 0.0005 Lauroyl-SDKP- OH N.B.: The same compounds as those described in FR 2 991 196 were used in the context of the present invention.

[0194] It has also been noticed possibilities for modifications of this formulation during the method for producing the neogouttes which is described below: [0195] the amount of water can be from 60% to 90% by weight relative to the total weight of the compounds involved, for example 77.2%; [0196] the amount of PEG 40 stearate can be from 5% to 20% by weight relative to the total weight of the compounds involved, for example 9.2%; [0197] the amount of phospholipids can be between 1.5% and 3% by weight relative to the total weight of the compounds involved, for example 1.75%; [0198] the amount of olive oil can be between 3% and 10% by weight relative to the total weight of the compounds involved, for example 4.49%; [0199] the amount of wax can be between 3% and 10% by weight relative to the total weight of the compounds involved, preferably of the same weight as the olive oil, for example 4.49%; [0200] the amount of palmitoyl-KTTKS can for example be between 0.01% and 10% by weight relative to the total weight of the compounds involved, for example 0.05%; [0201] the amount of lauroyl-SDKP-OH can for example be between 0.00001% and 1% by weight relative to the total weight of the compounds involved, for example 0.00006%.
A. Preparation of the aqueous phase

[0202] The aqueous phase was prepared by dissolving the Myrj S40 surfactant, dissolved in 1× phosphate buffered saline (PBS), in water.

B. Preparation of the Lipid Phase 30

[0203] The lipid phase was prepared by mixing soya bean oil (Soybean oil, Sigma Aldrich), paraffin (semi-synthetic glycerides, Suppocire NC, Gattefossé, France), soybean phospholipids (Phospholipon 75, Lipoid, Germany) and 0.1% by weight of Dil fluorophore (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate, Sigma Aldrich). The lipid phase thus prepared contains 16% by weight of phospholipids and 84% by weight of lipids.

C. Preparation of the neogouttes by ultrasonication

[0204] 20% of the lipid phase were dispersed in 80% of the aqueous phase, resulting in a mixture having a phospholipids/Myrj S40 ratio of 0.18 and a Myrj S40/(oil+wax) ratio of 0.55.

[0205] The emulsification of the mixture was then carried out with a 3 mm ultrasonic probe, according to (10 s ON/30 s OFF) sonication cycles for 10 min.

[0206] The nanoemulsion suspensions can then be dialyzed against 500 ml of 1× PBS overnight. They were then recovered, diluted to a content of 10% by weight, filtered through pores of 0.2 pm and, finally, stored at 4° C. until use.

Example 3: Dilution

[0207] It is possible to dilute neogouttes according to the present invention.

[0208] A hydrophilic solvent can be used for this purpose. Water is typically used. It is also possible to add preservatives, antimicrobial agents, or any other compound (e.g. excipient) commonly used in cosmetic, dermocosmetic and/or pharmaceutical formulations.

[0209] From a practical point of view, the neogouttes are diluted simply by adding a diluting solvent (e.g. water) to the concentrated formulation of neogoutte (as produced in example 2 above). The addition of the diluting solvent can be carried out with stirring (e.g. by a turbine), or the stirring can be carried out after the addition of the diluting solvent.

[0210] Thus, all the neogouttes presently described can be diluted in aqueous solutions, for example to ⅕.sup.th, 1/10.sup.th, 1/20.sup.th or else 1/50.sup.th (see procedure described in detail below).

[0211] For example, the neogouttes of example 2 can be diluted to 1/10.sup.th in an aqueous solution. An aqueous solution is then added with stirring to the formulation obtained according to example 2 until the initial solution is diluted to 1/10.sup.th (i.e. 9 volumes of aqueous diluting solution added to 1 volume of the formulation according to example 2).

Example 4: Comparative tests

[0212] In the context of the present invention, a comparative clinical test between neogouttes without lauroyl-SDKP-OH and neogouttes with lauroyl-SDKP-OH were carried out on healthy volunteers.

[0213] In the case of the formulations comprising the neogouttes without lauroyl-SDKP-OH, the missing portion of lauroyl-SDKP-OH in the neogouttes was filled with “Myrj S40”.

[0214] The amount of lauroyl SDKP-OH dosed into the neogouttes in the final product used in the context of these experiments was less than 1×10.sup.−5% by weight relative to the total weight of formulation.

[0215] Nevertheless, even at these levels of concentration, physiological effects were reported.

[0216] 1) Smoothing effect [0217] A significant smoothing effect was measured compared with placebo in 75% of cases as early as 14 days with an average roughness decreased by 11%. [0218] Variations up to −19%

[0219] 2) Anti-wrinkle effect [0220] A significant anti-wrinkle effect was measured compared with placebo in 81% of cases as early as 14 days and 77% at D28 with a variation in the relief of −14% and −16%, respectively. [0221] Variations up to −59%

[0222] 3) Restructuring/anti-aging effect [0223] A significant restructuring/anti-aging effect was measured in 81% of cases on D14 and 69% on D28 with an increase in the isotropy of 12% and 14%, respectively. [0224] Variations up to +28%. These results are summarized in FIG. 1.

[0225] 4) Redensifying effect [0226] A redensifying effect was measured in 68% of cases on D14 and 90% on D28, with an average increase in the skin density of respectively 12% and 37%. These results are summarized in FIG. 2.

[0227] 5) Effect on the dermal thickness [0228] A significant increase in the thickness of the dermis was measured in 72% of patients.

[0229] 6) Moisturizing effect [0230] A significant moisturizing effect was measured compared with placebo. [0231] Variations up to +20%

[0232] 7) Tonifying effect [0233] A significant improvement in the tonicity of the skin was measured in 53% of subjects. These results are summarized in FIG. 3.

[0234] These particularly encouraging results show the advantage of the choice of the technology according to patent FR 2 991 196 A applied to a modified formula of SDKP according to the present invention.