FR2753707A1 - NUCLEOPEPTIDIC COMPLEXES, COSMETOLOGICAL AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM AND USES - Google Patents

Abstract

The invention concerns nucleopeptide complexes obtained by synthesis, comprising a sequence of at least three amino acids in the form of D, L or DL, said sequence being chemically or physically bound with xanthic acids, or their derivatives of general formula I: N-R1 - COOH, in which N represents one purine base containing 5 to 10 carbon atoms, R1 represents one C1 or C3 alkyl radical.

Description

 The subject of the present invention is nucleo-peptide molecular complexes obtained by chemical synthesis, in the form of peptide conjugates, physically or chemically bound to purine acids, in particular xanthic acids and their derivatives.

 The physiological activity of the nucleo-peptides which are the subject of the present invention is preferentially directed towards the synthesis and stimulation of epidermal melanins and their dispersion at the level of the superficial layers of the skin - thus ensuring a very wide melanization of the epidermis and a high level of protection against solar radiation-induced "erythema" (UVA-UVB).

 It is known to those skilled in the art that the function of the melanocyte pigments, secreted by the cells of the human epidermis "melanocytes", as well as their dispersion at the keratinocyte cells of the skin, has the physiological function essential protection of the dermis, epidermis and immune system against solar radiation, especially UVA and UVB.

It is also known that the human melanocyte system (skin coloration, hair, etc.) functions differently from that of animals. In particular the secretion of melanins by melanocytes and the dispersion of these melanins on the surface of the epidermis requires in humans a cellular cooperation between melanocytes and keratinocytes, which is expressed by a defined number of melanocytes for a keratinocyte, thus forming a functional physiological entity called "Epidermal
Melanin Unit "specific to each racial type." Dubertret L. et al., J.

Invest. Dermatol. (8) suppl. 2-10-1983 - Dubertret L. et al. J. Invest.

Dermatol. 0022-202X-1986 - TB Fitzpatrick, G. Szabo, Michael M. Wick,
Biochemistry and Physiology of Melanin Pigmentation 687, vol. II, Lowell A.

Goldsmith, 1991 - Jimbow K, Uesugi T (1992), J. Invest. Dermato. 78-108-115,
Lerch K. (1988) Advances in Pigment Cell Research 85-88.

 The activity of the melanocyte system is dependent on physiological factors such as peptide hormones, growth factors, and in particular, induction and activation factors of melanocyte tyrosinase and cyclic AMP Protein Kinase dependent.

It is known to those skilled in the art that the cellular metabolism of the peptides, whether hormonal or otherwise (neurotransmitters or mediators), is under the control of the cell cycle of cyclic AMP, called the second hormone messenger, whose role is to amplify the cellular response induced by a specific peptide factor (Rall et al., J. Biol Chem 224-463 (1957)),
Sutherland, Rall. IBID 232-1077 (1958)).

 It has also been shown that certain substances are able to strongly potentiate the action of cyclic AMP thus prolonging Tyrosinase stimulation in the intracellular metabolism of melanogenesis.

 Among these substances are analogs of cyclic AMP such as dibutyryl cAMP, prostaglandins E1 and in particular xanthine derivatives (Halaban R, Pomerantz SH, Marshal S, Lambert DT and Lerner AB, J.

Cell. Biol. 97, 480-488 (1983); Kowichi, Jimbow, Thomas B. Fitzpatrick Mick, M.

Wick Biochemistry and Physiology of Melanin Oxfort Pigmentation U. Press (1991)).

 One of the objects of the present invention relates to molecular complexes in the form of nucleo-peptides comprising melanotropic peptide conjugates, specific inducers of cellular melanogenesis, chemically linked to puric acids or their derivatives, preferably to xanthic acids, stimulating melanosomal tyrosinase. by activation of cyclic AMP (2nd messenger) whose role is to amplify the activity of melanocyte cellular systems, and in particular the dispersion of melanins at the level of keratinocytes by activation of growth factors.

Among the peptide complexes composing the nucleo-peptides according to the invention, preferentially include peptides comprising a sequence of at least 3 amino acids, said amino acids being in D form,
L or DL, that is natural or not.

Said sequence is associated or linked chemically or physically, in the form of salts, esters or amides, with purine acids or their derivatives of general formula I:
N-R 1 -COOH (I) wherein N represents a purine base comprising 5 to 10 carbon atoms, R 1 represents a C 1 or C 3 alkyl radical.

 The acids of general formula (I) are chosen from purine or purine-derived acids for which R 1 represents a C 1 or C 3 alkyl radical, and are preferably chosen from: - hypoxanthic acid (C 7 H 5 N 4 O 3), - acid acetyl xanthine (C7H8N4O4) - theobromine acetic acid (C9H8N4O4), and - theophylline acetic acid (CgH11N4O4).

The peptide complexes of the invention preferably comprise a sequence of 3 amino acids, in particular one of the following sequences
Ala-His-HPhe
His-HPhe-Arg in which Hphé represents dihydroxy-DPhénylalanine or the
DhomoPhénylalanine.

In general, the peptide complexes according to the invention correspond to the general formula
AX-His-HPhé-Y in which:
A is a purine acid of general formula (I) as defined above
X represents a Norleucine (Nle) group, or one of its derivatives in the form
acetylated (Acyl-Me) or methylated (MeNle), Nle-Ala or Ala group
Y represents an ArgZ or Z group in which Z is Trp-Gly-OH, Trp-OH, OH,
Trp-Gly-NH2, Trp-NH2 or NH2;
Hphé represents dihydroxy-DPhénylalanine or DhomoPhénylalanine.

The present invention relates more particularly to the following peptide derivatives:
1) A-Nle-Ala-His-DhomoPhe-Arg-Trp-Gly-NH2
2) A-Nle-Ala-His-DhomoPhe-Arg-Trp-NH2
3) A-Nle-Ala-His-DhomoPhe-Arg-NH2
4) A-Nle-Ala-His-DhomoPhe-NH2
5) A-Ala-His-DhomoPhe-NH2
6) A-Nle-Ala-His-DhomoPhe-Arg-Trp-Gly-OH
7) A-Nle-Ala-His-DhomoPhe-Arg-Trp-OH
8) A-Nle-Ala-His-DhomoPhe-Arg-OH
9) A-Nle-Ala-His-DhomoPhe-OH
10) A-Ala-His-DhomoPhe-OH
11) A-Nle-Ala-His-DihydroxyDPhe-Arg-Trp-Gly-NH2
12) A-Nle-Ala-His-DihydroxyDPhe-Arg-Trp-NH2
13) A-Nle-Ala-His-DihydroxyDPhe-Arg-NH2
14) A-Nle-Ala-His-DihydroxyDPhe-NH2
15) A-Ala-His-DihydroxyDPhe-NH2
16) A-Nle-Ala-His-DihydroxyDPhe-Arg-Trp-Gly-OH
17) A-Nle-Ala-His-Dihydroxy-Pepe-Arg-Trp-OH
18) A-Nle-Ala-His-DihydroxyDPhe-Arg-OH
19) A-Nle-Ala-His-DihydroxyDPhe-OH
20) A-Ala-His-DihydroxyDPhe-OH.

 It is also possible that some of these amino acids have glycosylation functions.

 It should be understood that the present invention relates to all of these structures included in the present description.

 The present invention also relates to galenical, pharmaceutical or cosmetological compositions comprising, as active substance, at least one nucleopeptide complex as defined above, in association with a pharmaceutically or cosmetologically acceptable carrier.

 These compositions may be produced in the form of an emulsion, a cream, a spray, a solution, a lotion or a gel and may comprise known excipients as well as possibly other active ingredients.

The present invention advantageously comprises preparations for pharmaceutical use, usable by the injectable route or by the topical route, for the medical treatment of the disorders of melanogenesis, it is preferably peptide conjugates with carboxylic acids.
It also relates to cosmetic compositions and / or capillaries used in the form of solutions, lotions, sprays or creams or gels, used as an activator of cutaneous melanogenesis, in the absence of any erythema phenomenon, protecting thus the epidermis against trauma induced by solar radiation (UV). The cosmetological compositions according to the present invention can also be used for the preventive and curative treatment of solar erythemas.

 The invention will be better understood in view of the following examples which are given for purely illustrative purposes.

Example 1 Synthesis of Peptide N 1 - Code M1
A = Acetyl Xanthic Mu = 211.11
M1 = Peptide 1 = Nle - Ala - His - DhomoPhe - Arg - Trp - Gly - NH2
The syntheses of the peptide complexes according to the invention were carried out according to Merrifield techniques adapted to the different peptide structures of the complexes forming the subject of the invention.

 Since the general principle of the Merrifield technique is known to those skilled in the art, only the specific phases of each peptide sequence will be described.

 In general, an MBHA resin is used as protective group: the FMOC derivatives (Fluoroenyl, Methoxy Carbonyl), and as coupling agent: BOP.

The amino acids used in the form of FMOC derivatives are the following in the progressive order: FMOC-Gly-OH, Trp-OH, Arg-OH, DhomoPhé
OH, His-OH, Ala-OH and Nle-OH.

 Each amino acid is used in excess, as well as BOP and couplings repeated twice.

 Acetyl xanthine acid is coupled in a similar manner.

Deprotection is carried out in two phases
1 - trifluoroacetic acid,
2 - hydrofluoric acid anydre / para-cresol.

 The compound obtained with a yield of 75% is taken up in 95.5% acetic acid and desiccated by lyophilization.

Conjugate N "1 is obtained with a yield of 82% purity
HPLC.

 100 mg of this conjugate are again purified by HPLC on a C18 column and 70 mg of pure product is obtained.

Analytic
Retention time: 17 min, C18 column, UV 210 nm, triethylamine phosphate buffer solvent, acetonitrile.

 Amino acids: Ala - 1; Gly 1.01; His - 1.00; Arg - 0.94; Dh.Phé 1.03.

 Mass spectrum (FAB +) 1075.

Example 2 Synthesis of Peptide N 2 - Code M2
A = Theophylline acetic acid (CgH11N404)
M2 = MeNle-Ala-His-DhomoPhe-Arg-Trp-NH2
By operating according to the Merrifield techniques of Example No. 1, the M2 peptide is obtained after treatment with HPLC in a yield of 65%.

Analytic
MW = 1036
HPLC = 98% - Nucleosil C18 - 250
Amino acids: MeNle = ++, His = 1.03 / 1, Arg = 1.01 / 1, Ala = 0.96 / 1,
Dh-Phé = 1.05.

 ++ = MeNle = detected.

Example 3 Synthesis of Peptide N 4 - Code M4
A = acetic acid CH3COOH
MW = 60.05
M4 = Nle - Ala His - DHomo-Phe - NH2.

 By operating according to the techniques described above in Example N 1, the N 4 peptide is obtained.

Analytic
MW = 645
HPLC = 95% - Nucleosil C18 - 244
Amino acids: Me = ++, His = 1.03 / 1, Arg = 1.01 / 1, Ala = 0.96 / 1, Dh
Phé = 1.05.

 ++ = Me = detected.

Example 4 Synthesis of Peptide N 3 - Code M3
A = Theobromoacetic acid (C9H10N4O4)
M3 = Nle-Ala-His-DhomoPhe-Arg-NH2
Theobromoacetic acid is salified with the peptide. By operating according to the techniques described above in Example N 1, the N 3 peptide is obtained.

Analytic
MW Peptide = 745
MW Theobromoacetic = 238.20
3,7 dimethyl purine acetic
HPLC Peptide = 95%
Nucleosil C18 - 244
Amino acids: His = 1.05, Arg = 1 / 1.02, Ala = 0.96, HydroxyDh-Phe = 1.18.

 Nle = detected.

Example 5 In Vitro Study of Peptide M1 Activity on the
stimulation of melanin and tyrosinase, on
Melanocytes in culture compared with MS H MATERIEL
1. Peptide M1
PM = 1075
10-7 M; 10-9m; 10-11 M
2. MSH - Sigma
10-7 M
3. Cloudman Melanocytes - Clone M3.

METHOD
Cultures of Melanocytes in the Presence of Peptide M1 Solutions, and of αMSH as Control Culture Under 5% CO2 in Ham F.10 Medium Supplemented with Horse Serum (15%) and FCS (2.5%) in the Presence Penicillin-Streptomycin.

ASSAYS
= Determination of Melanine according to Abdel Malek at 436 nm.

 = Tyrosinase / Ldopa assay at 475 nm.

 = Protein assay according to Lowry technique.

RESULTS = Melanin assays
Peptide M1 causes an increase in the level of Melanine of 92% at a dilution of 10-7 M, and the effect of MSH is increased.
Melanin 11% at a dilution of 10-7 M.

RESULTS = Tyrosinase assays
% increase in melanin and tyrosinase

<tb><SEP>&alpha; MSH <SEP><SEP> M1 <SEP> M1 <SEP> M1
<tb><SEP> 10-7 <SEP> M <SEP> 10-7 <SEP> M <SEP> 10-9 <SEP> M <SEP> 10-11 <SEP> M
<tb><SEP> Melanin <SEP> 11 <SEP>% <SEP> 92 <SEP>% <SEP> 80 <SEP>% <SEP> 51 <SEP>%
<tb> Tyrosinase <SEP> 28 <SEP>% <SEP> 30 <SEP>% <SEP> 58 <SEP>% <SEP> 90 <SEP>%
<tb> COMMENTS
I - Cloudman's melanocytes
Cloudman's Melanoma murine melanocytes, whose property is to naturally secrete tyrosinase and melanin.

 The use of Cloudman's Melanoma for the primary screening of melanotropic substances lies in the exponential response of this experimental model, which makes it possible to differentiate the activities of melanotropic peptides whose structures are relatively close.

II - Dosage / effect relationships of melanotropic peptides
In the single-cell culture model, the response of peptides is dependent on the size of the cellular receptors that respond more intensively at low doses and spun.

 Moreover, there is a shift between the optimal melanotropic response of the peptide M1 at doses of 10-7 M and 10-9 M, and the response of the synthesis of Tyrosinase, which reaches its optimum at lllll.

 This phenomenon is explained by the release of melanin stored in melanosomes, while the synthesis of tyrosinase is induced after this release, to compensate for the secretion of melanin.

 Furthermore, the molecular weight of the peptides tested compared with the molecular weight of the control aMSH must be taken into account.

 For example, the aMSH at 10-7 M independently of its ubiquitous action, is not usable in therapy given the high cost of its synthesis which comprises 14 stages, while the peptide M1 comprises only 7 at the same power 10-7 M for an activity 9 times more intense.

Example 6 In Vitro Study of Tyrosinase Synthesis
induced in melanocytes in culture by the
M3 peptide Human melanocytes clone M4Be
INSERM U. 436 MATERTEL
1. Peptide M3
PM = 745
Used at dilutions of 10-7 M, 10-9 NI, 1011 M in M20.

2. AMSH Sigma reference reference peptide used at a dilution of 10-7 M
in H2O.

 3. Human melanocytes clone M4Be - INSERM U.436.

4. M4Be Melanocytes are tumor cells from Unity
U 436/218 of the INSERM of Lyon. These cells are characterized by their
Tyrosinase deficiency (Menkes syndrome).

 They represent an experimental model to assess the activity of peptides on the synthesis of Tyrosinase in genetically deficient subjects.

Culture
Witness aMSH Lot 085H12461
Peptide M3 Cells
-Type: S9lCD
- N passage: 21
- Inoculation, date: 72
- Inoculation, quantity: 1 x 10 5 / well, 3 plates 6 wells Medium
Ham F10 + (10% Sch, 2% FCS, Penis Strepto (100 U-100 μg / ml)).

Product contact (100 l + 0.9 ml medium):
- Duration: 24 hours
- Rinsing: 2
Cell recovery after: 24h
assays
Determination of Tyrosinase Ldopa at 475 nm.

 Protein Assay Using the Lowry Technique.

RESULTS Assay of tyrosine oxidase activity and proteins in melanocytes cultured in the presence of &alpha; -MSH or M3 peptide

Product <SEP> Tyrosinase <SEP> Proteins <SEP> Tyros./Prot. <SEP> Average <SEP> Standard Deviation <SEP> Stimulation
<tb> U / ml <SEP> g / ml <SEP> x <SEP> 1000 <SEP> Tyros./prot. <SEP> Tyros./prot. <SEP> in <SEP>%
<tb> 0.178 <SEP> 48.4 <SEP> 3.68
<tb> Control <SEP> 0.190 <SEP> 47.2 <SEP> 4.03 <SEP> 3.70 <SEP> 0.26 <SEP> /
<tb> 0.165 <SEP> 48.8 <SEP> 3.38
<tb> 0.225 <SEP> 48.91 <SEP> 4.60
<tb> MSH <SEP> (10-9 <SEP> M) <SEP> 0.237 <SEP> 47.12 <SEP> 5.03 <SEP> 4.63 <SEP> 0.54 <SEP> 25
<tb> 0.206 <SEP> 48.70 <SEP> 4.23
<tb> 0.287 <SEP> 66.6 <SEP> 4.31
<tb> 10-5 <SEP> M <SEP> 0.262 <SEP> 66.0 <SEP> 3.97 <SE> 4.23 <SE> 019 <SEP> 15
<tb> 0.266 <SEP> 60.3 <SEP> 4.41
<tb> 0.270 <SEP> 53.9 <SEP> 5.01
<tb> 10-7 <SEP> M <SEP> 0.245 <SEP> 56.8 <SEP> 4.31 <SEP> 4.67 <SEP> 0.28 <SEP> 27
<tb> 0.254 <SEP> 54.3 <SEP> 4.68
<tb> 0.304 <SEP> 46.3 <SEP> 6.57
<tb> M3 <SEP> 10-9 <SEP> M <SEP> 0.329 <SEP> 48.4 <SEP> 6.80 <SEP> 7.25 <SEP> 0.81 <SEP> 96
<tb> 0.371 <SEP> 44.2 <SEP> 8.39
<tb> 0.296 <SEP> 66.9 <SEP> 4.42
<tb> 10-11 <SEP> M <SEP> 0.207 <SEP> 49.7 <SEP> 4.16 <SEP> 4.29 <SEP> 0.11 <SEP> 16
<tb> 0.224 <SEP> 52.2 <SEP> 4.29
<tb> 0.241 <SEP> 57.0 <SEP> 4.23
<tb> 10-12 <SEP> M <SEP> 0.254 <SEP> 62.1 <SEP> 4.09 <SEP> 4.13 <SEP> 0.07 <SEP> 12
<tb> 0.283 <SEP> 69.5 <SEP> 4.07
<Tb>
Tyrosinase assays
M3 peptide stimulates tyrosinase secretion by 96% at a dose of 10-9 M compared to untreated controls and otMSH at a dose of 10-9 M.

Example 9 Study of the Preventive Power of Peptides M2 against
solar erythema
Goal:
To evaluate the protective effect of Peptide M2 in topical application against solar erythema.

MATERIALS
1. M2 peptide in solution at 50 mg / l and 100 mg / l in a PEG gel.

 2. Dunkin-Hartley male guinea pigs of 300 g, 10 animals per batch.

METHOD
Depilation of the flanks of the animals on D-1, one side receives the treatment, the other serves as a control. The animals are covered with metal paper pierced with 4 windows at the flanks. One side of 1 ml of Gel is applied to each of the M2 peptide solutions.

 After application, the animals are exposed to UVA and UVB radiation delivering 10.75 W / cm 2, each window being subjected to an exposure of increasing duration in geometric progression of 1.25.

 After 24 hours, the level of erythema of each window is evaluated.

RESULTS
For each animal, the irradiation time causing the first net erythema (DEM) is evaluated. The DEM ratios (treated area / untreated area) are then calculated.

The average of these ratios is the index of elevation of the erythema threshold for each dose.

<tb><SEP><SEP> Ranges of
<tb><SEP> Treatments <SEP> Time <SEP> DEM <SEP> Elevation <SEP> Indices
<tb><SEP> irradiation <SEP> average <SEP> of the <SEP> erythemal <SEP> threshold
<tb><SEP> Difference
<tb><SEP> no <SEP> controls processed <SEP> 60 <SEP> to <SEP> 117 <SEP> s <SE> 83.95 <SEP> Mean <SEP> type
<tb> M2 <SEP> 50 <SEP> mg / l <SEP> 240 <SEP> to <SEP> 468 <SEP> s <SEP> 324.40 <SEP> 3.96 <SE> 0.66
<tb> M2 <SEP> 100 <SEP> mg / l <SEP> 360 <SEP> to <SEP> 702s <SEP> 495.6 <SEP> 5.94 <SEP> 0.99
<Tb>

CONCLUSION
Application of the M2 peptide topically significantly prevents the appearance of solar erythema.

 Indeed, exposures 6 times (for 100 mg / l) and 4 times (for 50 mg / l) longer are necessary to give an erythema equivalent to that observed on the untreated part.

No signs of intolerance were observed in the areas that received
M2 then exposed to UV rays.

Example 10: Clinical study of objectification of the activity
comparative of two galenic formulas of the series
Peptide MELITANE
RESUME
To study comparatively the melantotropic and anti-erythematous action of two galenic formulas of peptides of the MEUTANE series, in topical application.

EQUIPMENT
Formula N "10 = A-Peptide N" 4
Acyl-MeNle-Ala-His-DhomoPhé-NH2
Formula N "11 = A-Peptide N" 3
3,7-dimethyl purine acetic acid-Nle-Ala-His-DhomoPhe-Arg-NH2
Galenic form = carboxymethyl cellulose gel with 5 mg peptide for
100 g of gel.

METHODOLOGY
The analysis of the results focused on a panel of 25 adult volunteers of both sexes, aged 18 to 50 years.

 The products studied were applied daily, twice a day, on the forearm flexion surface (one product per forearm) for two consecutive weeks.

 The statistical analysis concerned, on the one hand, the values obtained in each of the treated zones in comparison with the values obtained in the control zone and, on the other hand, on the calculated differences between the values obtained on each of the treated areas or control of the value obtained on the control zone specific to each of the different zones, in order to compare the two products studied with the control zone.

 The means of the chromaticity coordinate differences were also calculated for the whole of each of the treated and control areas.

 An analysis of variance and the least significant difference test (L.S.D.) also made it possible to make multiple comparisons between the values obtained in each of the treated and control zones.

 The mean values and the deviations obtained from the mean (SEM) of Minimum Erythematous Doses (DEM), obtained on all volunteers, for each of the products studied, are listed in Table I. The statistical differences are also mentioned, as well as the type of test performed.

 Mean values and deviations from the mean (SEM) of the differences in the chromaticity b * coordinate and the Individual Typological Angle (ITA), obtained on all the volunteers who took part in the test, for each of the products studied, are listed in Tables II, III, IV and V. The statistical differences are also mentioned, as well as the type of test performed.

 The Individual Typological Angle IOTA ", defined by Chardon & Col.

to determine the level of melanization of the skin, was calculated as follows: IOTA "= (arc tangent (L * -50) / b *) 180 / 3.14159 with: L * = lightness variable and b * = coordinate chromaticity from blue to yellow.

TABLE I
MINIMUM ERYTHEMATUS DOSES AFTER AN EXHIBITION
UV A AND B
(averages and SEM, n = 25)

<tb><SEP> DOSE <SEP> IN <SEP> J / cm2
<tb><SEP> ZONES <SEP> WITNESS <SEP> | <SEP> PRODUCT <SEP> 10 <SEP> PRODUCT <SEP> 11
<tb> T <SEP> 24 <SEP> hours <SEP> 1.5 <SEP>#<SEP> 0.1 <SEP> 1.8 <SEP>#<SEP><SEP> 0.1 * <SEP > 2.5 <SEP>#<SEP><SEP> 0.1 *
<tb> *: statistically significant value compared to the value obtained at
level of the control zone, at the same time of the study (Student's t-test
in unpaired series, one-tail, significance: p <0.05).

: statistically significant value compared to the value obtained at
level of the other treated area, at the same time of the study (LS.D. and test t
Student paired series, onetail, significance: p <0.05).

TABLE II
VARIATION OF THE COORDINATE OF CHROMATICITY b *
AFTER SOLAR EXPOSURE TO UV A AND B
T72 Hours
(averages and SEM, n = 25)

<tb><SEP># b *
<tb><SEP> Zones
<tb><SEP> FLUX <SEP> in <SEP> WITNESS <SEP> PRODUCT <SEP> 10 <SEP> PRODUCT <SEP> 11
<tb> med / min / cm2
<tb><SEP> 0.82 <SEP> 0.3 <SEP>#<SEP> 0.1 <SEP><SEP> 0.7 <SEP>#<SEP> 0.2 <SEP> 0.9 <SEP>#<SEP> 0.2
<tb><SEP> 1.02 <SEP> 0.4 <SEP>#<SEP> 0.1 <SEP><SEP> 0.9 <SEP>#<SEP> 0.2 <SEP><SEP> 1.4 <SEP>#<SEP> 0.2
<tb><SEP> 1.28 <SEP> 0.3 <SEP>#<SEP> 0.1 <SEP><SEP> 1.3 <SEP>#<SEP> 0.2 <SEP><SEP> 1.6 <SEP>#<SEP> 0.2
<tb><SEP> 1.60 <SEP> 0.5 <SEP>#<SEP> 0.1 <SEP><SEP> 1.5 <SEP>#<SEP> 0.2 <SEP><SEP> 1.9 <SEP>#<SEP> 0.2
<tb><SEP> 2.00 <SEP> 0.4 <SEP>#<SEP> 0.1 <SEP><SEP> 2.1 <SEP>#<SEP> 0.2 <SEP><SEP> 2.6 <SEP>#<SEP> 0.2
<tb><SEP> 2.50 <SEP> 0.5 <SEP>#<SEP><SEP> 0.1 <SEP> 2.5 <SEP>#<SEP> 0.2 <SEP><SEP> 2.9 <SEP>#<SEP> 0.2
<tb><SEP> AVERAGE <SEP> 0.4 <SEP>#<SEP> 0.1 <SEP> 1.5 <SEP>#<SEP> 0.2 * <SEP> 1.8 <SEP>#<SEP> 0.2 *
<tb> *: statistically significant difference with respect to the value obtained
at the level of the other treated area, for the same flow (ANOVA and LS.D.,
significance: p <0.05).

": statistically significant difference with respect to the value obtained
at the level of the control zone, for the same flow (Student's t test in
paired series, one-tail, significance: p <0.05).

TABLE III
VARIATION OF ITA AFTER SOLAR EXPOSURE TO UV A AND B
T24 Hours
(averages and SEM, n = 25)

<tb><SEP> A <SEP> I. <SEP> T. <SEP> A.
<Tb>

<SEP> Zones
<tb><SEP> FLOW <SEP> in <SEP> WITNESS <SEP> PRODUCT <SEP> 10 <SEP> PRODUCT <SEP> 11 <SEP>
<tb> Med / min / cm2 <SEP>
<tb><SEP> 0.82 <SEP> - <SEP> 1.5 <SEP>#<SEP> 0.3 <SEP> - <SEP> 2.8 <SEP>#<SEP> 0.6 <SEP> - <SEP><SEP> 3,6 <SEP>#<SEP> 0,5
<tb><SEP> 1.02 <SEP> - <SEP> 1.9 <SEP>#<SEP> 0.7 <SEP><SEP> - <SEP> 4.7 <SEP>#<SEP> 0 , 6 <SEP> - <SEP><SEP> 5.8 <SEP>#<SEP> 0.7
<tb><SEP> 1.28 <SEP> - <SEP> 1.4 <SEP>#<SEP> 0.6 <SEP><SEP> - <SEP> 4.9 <SEP>#<SEP> 0 , 5 <SEP><SEP> - <SEP> 5.8 <SEP>#<SEP> 0.6
<tb><SEP> 1.60 <SEP> - <SEP> 1.7 <SEP>#<SEP> 0.6 <SEP><SEP> - <SEP> 5.5 <SEP>#<SEP> 0 , 7 <SEP><SEP> - <SEP> 7.0 <SEP>#<SEP> 0.7
<tb><SEP> 2.00 <SEP> - <SEP> 1.9 <SEP>#<SEP> 0.7 <SEP> - <SEP><SEP> 6.8 <SEP>#<SEP> 0 , 8 <SEP><SEP> - <SEP> 7.5 <SEP>#<SEP> 0.9
<tb><SEP> 2.50 <SEP> - <SEP> 2 <SEP>#<SEP> 0.7 <SEP><SEP> - <SEP> 7.5 <SEP>#<SEP> 1.0 <SEP><SEP> - <SEP> 8 <SEP>#<SEP> 0.9
<tb><SEP> AVERAGE <SEP> - <SEP> 1.75 <SEP> * <SEP> 0.6 <SEP> - <SEP> 5.37 <SEP>#<SEP><SEP> 0.7 * 0 <SEP> - <SEP> 6.29 <SEP> * <SEP> 0.72 * <SEP>
<tb> *: statistically significant difference from the value obtained
at the level of the other treated area, for the same flow (ANOVA and LS.D.,
significance: p <0.05).

": statistically significant difference with respect to the value obtained
at the level of the control zone, for the same flow (Student's t test in
paired series, one-tail, significance: p <0.05).

TABLE IV
VARIATION OF ITA AFTER SOLAR EXPOSURE TO UV A AND B
T72 Hours
(averages and SEM, n = 25)

<tb><SEP>#<SEP> I. <SEP><SEP> T. <SEP> A.
<tb><SEP> Zones
<tb><SEP> FLUX <SEP> in <SEP> WITNESS <SEP> PRODUCT <SEP> 10 <SEP> PRODUCT <SEP> 11
<tb><SEP> Med / min / cm2
<tb><SEP> 0.82 <SEP> - <SEP> 1 <SEP>#<SEP> 0.4 <SEP><SEP> - <SEP> 2.5 <SEP>#<SEP> 0.6 <SEP><SEP> - <SEP> 3 <SEP>#<SEP> 0.7
<tb> 1.02 <SEP> - <SEP> 1 <SEP>#<SEP><SEP> 0.5 <SEP> - <SEP> 3.3 <SEP>#<SEP> 0.6 <SEP> - <SEP> 5 <SEP>#<SEP> 0.8
<tb><SEP> 1.28 <SEP> - <SEP> 1 <SEP>#<SEP> 0.5 <SEP><SEP> - <SEP> 4.4 <SEP>#<SEP> 0.7 <SEP><SEP> - <SEP> 6 <SEP>#<SEP> 1.0
<tb><SEP> 1.60 <SEP> - <SEP> 2 <SEP>#<SEP> 0.5 <SEP><SEP> - <SEP> 5.5 <SEP>#<SEP> 0.9 <SEP><SEP> - <SEP> 7 <SEP>#<SEP> 0.8
<tb><SEP> 2.00 <SEP> - <SEP> 2 <SEP>#<SEP> 0.6 <SEP> - <SEP><SEP> 7.9 <SEP>#<SEP> 0.8 <SEP><SEP> - <SEP> 8 <SEP>#<SEP> 0.9
<tb><SEP> 2.50 <SEP> - <SEP> 3 <SEP>#<SEP> 0.5 <SEP><SEP> - <SEP> 8.1 <SEP>#<SEP> 0.9 <SEP><SEP> - <SEP> 10 <SEP>#<SEP> 0.7
<tb><SEP> AVERAGE <SEP> - <SEP> 1.66 <SEP> t <SEP> 0.5 <SEP> - <SEP> 5.29 <SE> s <SEP> 0.75 * <SEP > - <SEP> 6.5 <SEP> t <SEP> 0.82 * <SEP>
<tb> *: statistically significant difference from the value obtained
at the level of the other treated area, for the same flow (ANOVA and LS.D.,
significance: p <0.05).

statistically significant difference from the value obtained
at the level of the control zone, for the same flow (Student's t test in
paired series, one-tail, significance: p <0.05).

TABLE V
VARIATION OF THE COORDINATE OF CHROMATICITY b * and ITA '
ON TIME 72 Hours
(averages and SEM, n = 25)

<tb><SEP> DOSE <SEP> EQUAL <SEP> A <SEP> 0.75 <SEP> D.EM. <September>
<Tb>

<SEP> PARAMETER <SEP> INDICATOR <SEP> PRODUCT <SEP> 10 <SEP> PRODUCT <SEP> 11
<tb><SEP>#b<SEP><SEP> 0.5 <SEP>#<SEP> 0.1 <SEP><SEP> 2.5 <SEP>#<SEP> 0.1 * <SEP><SEP> 3.8 <SEP>#<SEP> 0.2 *
<tb><SEP> L <SEP> L <SEP> T. <SEP> A. <SEP> - <SEP> 2.2 <SEP>#<SEP> 0.4 <SEP><SEP> - <SEP > 3.5 <SEP>#<SEP><SEP> 0.4 * <SEP> - <SEP> 4.9 <SEP> t <SEP> 0.6 * <SEP>
<tb><SEP> DOSE <SEP> EQUIVALENT <SEP> A <SEP> 1 <SEP> DM <SEP>
<Tb>

<SEP> PARAMETER <SEP> INDICATOR <SEP> PRODUCT <SEP> 10 <SEP> PRODUCT <SEP> 11
<tb><SEP>#b<SEP><SEP> 0.6 <SEP>#<SEP> 0.1 <SEP><SEP> 1.7 <SEP>#<SEP> 0.2 * <SEP><SEP> 2.9 <SEP>#<SEP> 0.2 *
## EQU1 ## SEP>#<SEP><SEP> 0.6 * <SEP> - <SEP> 3.4 <SEP>#<SEP><SEP> 0.6 *
<tb><SEP> DOSE <SEP> EQUIVALENT <SEP> A <SEP> 1.25 <SEP> D.EM.
<Tb>

<SEP> PARAMETER <SEP> INDICATOR <SEP> PRODUCT <SEP> 10 <SEP> PRODUCT <SEP> 11
<tb><SEP>#b<SEP><SEP> 0.8 <SEP>#<SEP> 0.1 <SEP> 1.9 <SEP>#<SEP> 0.2 * <SEP> 2.6 <SEP>#<SEP> 0.2 *
<tb><SEP> A <SEP> I. <SEP> T. <SEP> A. <SEP> - <SEP> 3.8 <SEP>#<SEP> 0.6 <SEP><SEP> - <MS> 2.6 <SEP> t <SEP> 0.7 * <SEP> - <SEP> 1.5 <SEP> t <SEP> 0.6 * <SEP>
<tb> *: statistically significant value compared to the value obtained at
level of the control zone, at the same time of the study (Student's t-test
in paired series, one-tail, significance: p <0.05).

:: statistically significant value compared to the value obtained at
level of the other treated area, at the same time of the study (ANOVA and
LS.D., significance: p <0.05).

RESULTS
Topical applications of formula 10 and formula 11 products for 15 consecutive days and under the experimental conditions defined by the protocol gave the following results:
1. Highly statistically significant increase in dose
minimal erythematous in the adult volunteer, expressing an activity
anti-erythematous highly significant especially for the
formula 11.

2. There is also an increase for all panellists
statistically significant chromaticity indices reflecting a
high level elevation of the degree of melanization of the skin after
unique solar exposure to UV A and B.

3. The highly significant decrease in ITA also confirms
melanization intensity of the skin particularly marked for the
formula 11.

Claims (17)

 Synthetically synthesized nucleopeptide complexes comprising a sequence of at least three amino acids in D, L or DL form, said sequence being physically or chemically bound to purine acids, or their derivatives of general formula I:  N-R 1 -COOH (I) wherein N represents a purine base comprising 5 to 10 carbon atoms, R 1 represents a C 1 or C 3 alkyl radical.  2 / nucleopeptide complexes according to claim 1, characterized in that the derivatives of purine acids are xanthine acids.  3 / nucleo-peptide complexes according to claim 1 or 2, characterized in that the amino acid sequence is linked to the acid in the form of salt, ester or amide.  4 / nucleopeptide complexes according to one of claims 1 to 3, characterized in that the purine acids are chosen from the group comprising:  - hypoxantic acid (C7H5N403)  - acetyl xanthine acid (C7H8N4O4)  theobromine acetic acid (C9H8N4O4)  theophylline acetic acid (C9H11N4O4).  5 / nucleopeptide complexes according to one of claims 1 to 4, characterized in that the sequence consists of 3 amino acids, and is selected from the group consisting of:  Ala-His-HPhe  His-HPhe-Arg where Hphé represents dihydroxy-DPhénylalanine, DhomoPhénylalanine.  6 / nucleopeptide complexes according to claim 5, characterized in that they correspond to the general formula  AX-His-HPhé-Y in which A is a purine acid of general formula I as defined above; X represents a Norleucine (Nle) group, or one of its derivatives in the form  cetyl (Acyl-Me) or methylated (MeNle), the Nle-Ala or Ala group Y represents an ArgZ or Z group in which Z is Trp-Gly-OH, Trp-OH, OH,  rp-Gly-NH2, Trp-NH2 or NH2; Hphé represents dihydroxy-DPhénylalanine or DhomoPhénylalanine.  7 / nucleo-peptide complexes according to claim 6, characterized in that they are chosen from the group comprising  1) A-Nle-Ala-His-DhomoPhe-Arg-Trp-Gly-NH2  2) A-Nle-Ala-His-DhomoPhe-Arg-Trp-NH2  3) A-Nle-Ala-His-DhomoPhe-Arg-NH2  4) A-Nle-Ala-His-Dhomophé-NH2  5) A-Ala-His-DhomoPhe-NH2  6) A-Nle-Ala-His-DhomoPhe-Arg-Trp-Gly-OH  7) A-Nle-Ala-His-DhomoPhe-Arg-Trp-OH  8) A-Nle-Ala-His-DhomoPhe-Arg-OH  9) A-Nle-Ala-His-DhomoPhe-OH  10) A-Ala-His-DhomoPhe-OH  11) A-Nle-Ala-His-Dihy droxyDPhe-Arg-Trp-Gly-NH2  12) A-Nle-Ala-His-DihydroxyDPhe-Arg-Trp-NH2  13) A-Nle-Ala-His-DihydroxyDPhe-Arg-NH2  14) A-Nle-Ala-His-DihydroxyDPhe-NH2  15) A-Ala-His-DihydroxyDPhe-NH2  16) A-Nle-Ala-His-DihydroxyDPhe-Arg-Trp-Gly-OH  17) A-Nle-Ala-His-Dihydroxy-Pepe-Arg-Trp-OH  18) A-Nle-Ala-His-DihydroxyDPhe-Arg-OH  19) A-Nle-Ala-His-DihydroxyDPhe-OH  20) A-Ala-His-DihydroxyDPhe-OH.  8 / nucleopeptide complexes according to one of claims 1 to 7, characterized in that at least one amino acid constituting the sequence is glycosylated.  9 / cosmetological or pharmaceutical compositions, characterized in that they comprise, as active substance, at least one nucleopeptide complex according to one of claims 1 to 8, in association with a pharmaceutically acceptable carrier.  10 / cosmetological or pharmaceutical compositions according to claim 9, characterized in that they are in the form of solutions, lotions, sprays, creams or gel.  11 / Pharmaceutical compositions according to claim 9, characterized in that they are in the form of injectable preparations.  12 / Use of the cosmetological compositions according to claim 9 or 10, as an activator of cutaneous melanogenesis.  13 / Use of the complexes according to one of claims 1 to 8 for the preparation of a pharmaceutical composition according to one of claims 9 to 1 1 for the medical treatment of disorders of melanogenesis.  14 / Cosmetic compositions, characterized in that they comprise, as active substance, at least one nucleopeptide complex according to one of claims 1 to 8, in combination with a cosmetologically acceptable carrier.  15 / cosmetological compositions according to claim 14, characterized in that they are in the form of solutions, lotions, sprays, creams or gel.  16 / Use of cosmetological compositions according to claim 14 or 15, topically for the acceleration of the melanogenesis of the skin and hair.  17 / Use of cosmetological compositions according to claim 14 or 15, topically for the preventive and curative treatment of solar erythemas.