Title:  Anti-inflammatory compositions comprising peptide derivatives of .alpha.-MSH/algal extracts

United States Patent:  6,337,315

Assignee:  Societe L'Oreal S.A. (Paris, FR)

Filed:  July 15, 1999

Foreign Application Priority Data:  Jul 15, 1998[FR] (98-09055)

Anti-inflammatory compositions, well suited for a wide variety of therapeutic/cosmetic applications, comprise combinatory immixture of (1) an effective anti-inflammatory amount of at least one peptide derivative of .alpha.-type melanocyte stimulating hormone (.alpha.-MSH), or functional biological equivalent thereof, and (2) an effective anti-inflammatory response-enhancing amount of at least one marine algal extract.

DETAILED DESCRIPTION OF BEST MODE AND SPECIFIC/PREFERRED EMBODIMENTS OF THE INVENTION

More particularly according to the present invention, the peptide derivative is a derivative of .alpha.-type melanocyte stimulating hormone (.alpha.-MSH) or melanotropin. While .alpha.-MSH was originally described as being produced by the pituitary gland, the brain in general, the blood, the skin and other tissues are also able to produce .alpha.-MSH.

Thus, it has been shown by Schauer et al. (J. Clin. Invest., 93, May 1994 pp. 2258-2262) that the keratinocytes of the epidermis are a source of .alpha.-MSH. Receptors for .alpha.-MSH are present in a large number of cell types, in particular in the hair follicles of the human scalp (Pigment Cell Res., 4:193-8, 1991).

(1-13) .alpha.-MSH is known for its antipyretic activity, its anti-inflammatory activity and its pigmentation-promoting activity. This neuropeptide is recognized for inhibiting the inflammation which is induced by cytokines and other inflammation mediators, as well as by irritants.

The antipyretic signal of .alpha.-MSH is situated at its carboxy terminal sequence and can be mimicked by the 11-13 carboxy terminal tripeptide (L)Lys(L)Pro(L)Val (Watanabe et al., Brain Research Bulletin., Vol. 32. pp. 311-314, 1993).

Thus, U.S. Pat. No. 5,028,592 and WO-88/00833 describe the use of the tripeptide (L or D)Lys-(L)Pro-(L or D)Val in an anti-inflammatory therapeutic treatment and the preparation of such a drug for treating inflammation.

Other derivatives of .alpha.-MSH are recognized for their anti-inflammatory activity. For example, WO-95/08564, hereby expressly incorporated by reference, describes the anti-inflammatory activity of compounds which comprise at least one 4-amino acid sequence from .alpha.-MSH conjugated to thioctic acid.

The following compounds I to VII described in WO-95/08564 are more specifically representative:

I. [(DL)Lip] Glu-His-D.homoPhe-Arg-Trp-Gly-NH₂

II. [(DHLip] Glu-His-D.homoPhe-Arg-Trp-Gly-NH₂

III.[(DL)Lip] Glu-His-ParaFluoroPhe-Arg-Trp-Gly-NH₂

IV. [(DL)Lip] His-D.homoPhe-Arg-Trp-NH₂

V. [N.lipoyl-Lysine] Glu-His-D.homoPhe-Arg-Trp-Gly-NH₂

VI. [N.lipoyl-Lysine] His-D.homoPhe-Arg-Trp-Gly-NH₂

VII.[N.lipoyl-Lysine] His-D.homoPhe-Arg-Trp-NH₂

as well as the derivatives thereof, in particular the salts, esters or amides.

It should be appreciated that SEPORGA markets a product under the trademark MODULENE.RTM. which is a peptide derivative of .alpha.-MSH and which exhibits anti-inflammatory properties.

Nonetheless, it has now surprisingly and unexpectedly been determined that the anti-inflammatory properties of derivatives of .alpha.-MSH can be markedly enhanced by formulating these derivatives in combinatory immixture with an algal extract of marine origin.

Thus, it has now been demonstrated that intimately admixing a peptide derivative of .alpha.-MSH with an algal extract of marine origin exhibits an anti-inflammatory effect which is greater than the simple addition of the anti-inflammatory effects which these active agents exhibit when administered individually. Furthermore, it too has been demonstrated that the immixture elicits an anti-inflammatory response, while each of the constituents of the subject compositions, at concentrations at which, when administered alone, no therapeutic activity is observed.

Thus, other than the advantage that the combinatory formulation exhibits an anti-inflammatory response which is greater than that of the active agents considered individually, the immixture also makes it possible to incorporate each of the components thereof at concentrations which are lower than those required for each of the active agents when administered alone.

The examples presented below illustrate these particular features.

Accordingly, this invention features anti-inflammatory compositions which comprise, as the active principles thereof, an effective amount of at least one peptide derivative of .alpha.-MSH, or any functional biological equivalent, and at least one algal extract of marine origin.

By "functional biological equivalent" is intended a peptide which is functionally equivalent in terms of biological function and at least one of whose amino acid residues has been exchanged for an amino acid residue having a similar hydropathic index.

The hydropathic index is an index which is assigned to amino acids in accordance with their hydrophobicity and their charge (Kyte et al., J. Mol. Biol., 157:105 (1982)).

Among amino acids, the geometry of these molecules is such that they can theoretically appear in the form of different optical isomers. Thus, there is a molecular conformation of the amino acid (aa) which is such that it turns the plane of polarization of light to the right (dextrorotatory conformation or D-aa) and a molecular conformation of the amino acid (aa) which is such that it turns the plane of polarization of light to the left (laevorotatory conformation or L-aa).

Nature has maintained only the laevorotatory conformation for the natural amino acids. Consequently, if the peptide employed in the compositions according to the invention is of natural origin, this peptide will comprehend amino acids of the L-aa type. However, chemical synthesis in the laboratory makes it possible to prepare amino acids possessing the two possible conformations. Proceeding from this base material, it is possible to incorporate amino acids in the form of dextrorotatory or laevorotatory optical isomers equally well during the peptide synthesis. It is thus possible, during the peptide synthesis, to incorporate the amino acid residues Lysine-Proline-Valine equally well in their D-Lysine (D-Lys), L-Lysine (L-Lys), D-Proline (D-Pro), L-Proline (L-Pro), D-Valine (D-Val) or L-Valine (L-Val) form.

Hence, the peptide derivatives of the invention can be peptides whose amino acid residues are equally well in the form of dextrorotatory or laevorotatory optical isomers.

The peptides containing at least one of the following tripeptides are exemplary:

D-Lys-D-Pro-D-Val,

D-Lys-D-Pro-L-Val,

D-Lys-L-Pro-D-Val,

L-Lys-D-Pro-D-Val,

D-Lys-L-Pro-L-Val,

L-Lys-D-Pro-L-Val,

L-Lys-L-Pro-D-Val,

L-Lys-L-Pro-L-Val.

It will of course be appreciated that, consistent herewith, it is possible to formulate more than one peptide. In this event, the peptide mixture can be one of the possible combinations of the peptides indicated above.

It could transpire that, for reasons of resistance to degradation, according to the invention, a protected form of the peptide is required. The form of the protective group should obviously be a form which is biologically compatible. A large number of biologically compatible protective groups are available, such as, for example, acylation or acetylation of the amino terminal endgroup and/or amidation of the carboxy terminal endgroup.

Thus, the peptides of the invention can be peptides which are or are not in protected form.

Preferred protective groups are those based on acylation or acetylation of the amino terminal and/or on amidation of the carboxy terminal endgroup.

In particular according to the invention, the peptide derivative of .alpha.-MSH is selected from among the peptide derivatives comprising at least the tripeptide Lys-Pro-Val, the peptide derivatives comprising at least one 4-amino acid sequence from .alpha.-MSH which is or is not conjugated to thioctic acid, and, more preferably, the compounds described in WO-95/08564.

Preferred are the following compounds I to VII:

I [(DL)Lip] Glu-His-D.homoPhe-Arg-Trp-Gly-NH₂

II [(DH Lip] Glu-His-D.homoPhe-Arg-Trp-Gly-NH₂

III [(DL)Lip] Glu-His-ParaFluoroPhe-Arg-Trp-Gly-NH₂

IV [(DL)Lip] His-D.homoPhe-Arg-Trp-NH₂

V [N.lipoyl-Lysine] Glu-His-D.homoPhe-Arg-Trp-Gly-NH₂

VI [N.lipoyl-Lysine] His-D.homoPhe-Arg-Trp-Gly-NH₂

VII [N.lipoyl-Lysine] His-D.homoPhe-Arg-Trp-NH₂

as well as the derivatives of these molecules, e.g., the salts, esters or amides thereof.

A peptide derivative comprising at least the tripeptide Lys-Pro-Val and which is preferred according to this invention is the tripeptide Lys-Pro-Val itself and, more particularly, the tripeptide Lys-Pro-Val in which at least the Proline amino acid residue is in the unnatural dextrorotatory conformation (DPro residue).

Another derivative which is preferred according to this invention is the derivative marketed by Seporga under trademark Modulene.RTM..

The peptide in accordance with the invention can, of course, be of natural origin. This implies that it can have been purified from natural biological material. It is possible, in this regard, to note, by way of example, .alpha.-MSH, which is widely present in the central nervous system and which can, inter alia, be purified from pituitary glands. However, given the current progress in chemical genetics, it is now quite easy to synthesize peptides, even of substantial length, to order.

Thus, the peptide derivatives of the invention can be peptides of natural or synthetic origin.

In the compositions of the invention, the peptide derivative can be a mixture of peptide derivatives.

The algal extract of marine origin can be any algal extract of marine origin, whatever the process by which it is obtained, with the reservation that it corresponds to the criterion selected for the invention, namely, that of exerting an anti-inflammatory enhancing effect on the anti-inflammatory activity of the peptide derivative.

Preferably, the algal extract of marine origin is an extract of brown algae of the Laminaria family. Even more preferably, the brown alga is an alga of the species Laminaria digitata.

An extract which is particularly preferred is an oligosaccharide solution which is obtained by enzymic depolymerization of brown algal membrane polysaccharides, such as described, in particular, in FR-2,753,628, hereby expressly incorporated by reference.

In this respect, an algal extract of marine origin which is particularly preferred according to the invention is an extract marketed by CODIF INTERNATIONAL under the designation PHYCOSACCHARIDES ANTI-IMNFLAMMATION.RTM. and which is a concentrated solution of an oligosaccharide which is obtained by the controlled enzymic depolymerization of membrane polysaccharides of a brown alga. It comprises the sequence of two uric acids: mannuronic acid and guluronic acid.

The compositions of the invention are of course compositions which are destined for cosmetic or pharmaceutical application.

The amount of each of the components of the admixture in accordance with the invention depends, quite obviously, on the desired effect and should be an amount which is effective for ensuring that the mixture elicits the desired effect, in particular an anti-inflammatory response.

To provide an order of magnitude the compositions of the invention advantageously comprise the peptide derivative in an amount by weight which represents from 10^-6 % to 10% of the total weight of the composition, and preferably in an amount which represents from 10^-3 % to 5% of the total weight of the composition.

Similarly, also to provide an order of magnitude, the compositions of the invention advantageously comprise the algal extract in an amount by weight which represents from 0.01% to 10% of the total weight of the composition, and preferably in an amount which represents from 0.02% to 5% of the total weight of the composition.

The invention also features formulating, as the active principle, of immixture of an effective amount of at least one peptide derivative of .alpha.-MSH, or any functional biological equivalent thereof, with at least one algal extract of marine origin into compositions well suited for treating inflammation.

This invention also features formulating, as the active principle, of immixture of an effective amount of at least one peptide derivative of .alpha.-MSH, or any functional biological equivalent thereof, with at least one algal extract of marine origin into compositions well suited for partially or totally inhibiting the production of interleukin 8, in particular by the keratinocytes of the skin.

Examples of disorders which manifest an inflammatory component are those indicated above.

Such inflammatory disorders may be skin disorders or systemic disorders.

Thus, the compositions comprising at least one peptide derivative of .alpha.-MSH and at least one algal extract of marine origin according to the invention are well suited for controlling disorders which manifest an inflammatory component, more specifically skin disorders.

In particular, the compositions according to the invention are useful for controlling skin ailments and afflictions such as sensitive skins, skin discomfort, skin stretching, skin itching, skin swelling, skin pain, skin flushing, heat sensation of the skin, erythemas, especially those due to ultraviolet rays, pruritus, erythema nodosum, urticaria, insect bites, allergies and alopecia in its inflammatory phases.

Even more particularly, the compositions according to the invention are useful for controlling skin irritations and/or sores and/or dysaesthesic sensations and/or heating sensations and/or prurituses of the skin and/or the mucous membranes.

Whatever the embodiment of the invention, the subject compositions can be ingested, injected or topically applied to the skin (on any region of the skin of the body), the hair, the nails or the mucous membranes (oral, jugal, gingival, genital and conjunctival). Depending on the mode of administration, the compositions according to the invention can be formulated into any of the galenic forms which are conventional in this art, e.g., formulated into any suitable medium, whether carrier, diluent or vehicle therefor.

For topical application onto the skin, the subject compositions can have the form, in particular, of an aqueous or oily solution or of a dispersion of the lotion or solution type, of emulsions of liquid or semi-liquid consistency of the milk type, which are obtained by dispersing a fatty phase in an aqueous phase (O/W) or vice versa (W/O), or of suspensions or emulsions of soft consistency of the aqueous or anhydrous cream or gel type, or else of microcapsules or microparticles or of vesicular dispersions of the ionic and/or non-ionic type. These compositions are formulated in accordance with the usual techniques. They can also be employed for the hair in the form of aqueous, alcoholic or hydroalcoholic solutions, or in the form of creams, gels, ointments, emulsions or mousses, or else in the form of aerosol compositions which also comprise a propellant under pressure.

For injection, the compositions can be in the form of an aqueous or oily lotion or in the form of a solution. For the eyes, the compositions are advantageously in the form of drops and, for ingestion, in the form of capsules, granules, syrups or tablets.

The amounts of the different constituents of the compositions according to the invention are those which are normally employed in the fields under consideration.

These compositions constitute, in particular, cleansing creams, protection creams, treatment creams or care creams for the face, for the hands, for the feet, for the large anatomical folds or for the body (for example, day creams, night creams, makeup removing creams, foundation creams and sunscreen creams), foundation liquids, makeup removing milks, body milks for protection or care, after-sun milks, lotions, gels or mousses for the care of the skin, such as cleansing lotions, sunscreen lotions, lotions for artificial tanning, bath compositions, deodorant compositions which comprise a bactericidal agent, after-shave gels or lotions, depilatory creams, compositions for treating insect bites, pain-relief compositions, and compositions for treating certain disorders of the skin such as eczema, rosacea, psoriasis, lichens and severe prurituses.

The compositions according to the invention can also be formulated as solid preparations constituting soaps or cleansing bars.

The subject compositions can also be packaged in the form of an aerosol composition which also comprises a propellant under pressure.

The compositions according to the invention can also be formulated for hair care, in particular a shampoo, a hair-setting lotion, a treatment lotion, a hair-styling cream or gel, a dye composition (in particular oxidation dyes), where appropriate in the form of coloring shampoos, restructuring lotions for the hair, a permanent-waving composition (in particular a composition for the initial stage of a permanent-waving operation), a lotion or gel preventing hair loss, an antiparasitic shampoo, etc.

The subject compositions are also useful for oral/dental application, for example formulated as a toothpaste. In this case, the compositions can contain adjuvants and additives which are customary for compositions for oral use, in particular surface-active agents, thickening agents, moistening agents, polishing agents such as silica, various active ingredients such as fluorides, in particular sodium fluoride, and, where appropriate, sweeteners such as sodium saccharinate.

When the composition is an emulsion, the proportion of the fatty phase advantageously ranges from 5% to 80% by weight, preferably from 5% to 50% by weight, the total weight of the composition. The oils, the waxes, the emulsifiers and the coemulsifiers employed in the compositions in emulsion form are selected from those which are customary in the cosmetic field. The emulsifier and the coemulsifier are advantageously present in the composition in a proportion ranging from 0.3% to 30% by weight, preferably from 0.5% to 20% by weight, of the total weight of the composition. The emulsion may furthermore contain lipid vesicles.

When the composition is an oily solution or gel, the fatty phase can represent more than 90% of the total weight of the composition.

In known manner, the cosmetic compositions may also contain additives and adjuvants which are customary in the cosmetic field, such as hydrophilic or liporhilic gelatinizing agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, perfumes, fillers, filters, sequestering agents, odor absorbents and dyes and colorants. The amounts of these different additives and adjuvants are those which are normal in the cosmetic field, for example from 0.01% to 10% of the total weight of the composition. Depending on their nature, these additives and adjuvants can be introduced into the fatty phase, into the aqueous phase and/or into the lipid spherules. Exemplary oils or waxes which are useful according to the invention, include mineral oils (liquid paraffin), vegetable oils (liquid fraction of karite butter, sunflower oil), animal oils (perhydrosqualene), synthetic oils (Purcellin's oil), siliconated oils or waxes (cyclomethicone) and fluorinated oils (perfluoropolyethers), beeswax and carnauba wax, or paraffin. Fatty alcohols and fatty acids (stearic acid) can be added to these oils.

Exemplary emulsifiers which are useful according to the invention include glycerol stearate, polysorbate 60, and the mixture of PEG-6/PEG-32/glycol stearate which is marketed by Gattefosse under the trademark Tefose R 63.

Exemplary solvents according to the invention include lower alcohols, in particular ethanol, isopropanol and propylene glycol.

And exemplary hydrophilic gelatinizing agents according to the invention include carboxyvinyl polymers (carbomer), acrylic copolymers such as acrylate/alkyl acrylate copolymers, polyacrylamides, polysaccharides such as hydroxypropylcellulose, natural gums and clays, and lipophilic gelatinizing agents include are modified clays such as bentones, metallic salts of fatty acids such as aluminum stearates, and hydrophobic silica, ethylcellulose and polyethylene.

The subject compositions can contain other hydrophilic active compounds such as proteins or protein hydrolysates, amino acids, polyols, urea, allantoin, sugars and sugar derivatives, water-soluble vitamins, plant extracts and hydroxy acids.

Exemplary lipophilic active compounds include retinol (vitamin A) and derivatives thereof, tocopherol (vitamin E) and derivatives thereof, essential fatty acids, ceramides, essential oils, and salicylic acid and derivatives thereof.

The present invention also features a cosmetic treatment regime or regimen, in particular with a view to decreasing inflammation, which treatment is characterized in that a composition as described above is topically applied to the skin, to the hair and/or to the mucous membranes.

Too, this invention features, in particular, a cosmetic treatment with a view to achieving a soothing effect on the skin, characterized in that a composition as described above is topically applied to the skin, to the hair and/or to the mucous membranes.

The cosmetic treatments of the invention can be implemented, in particular, by topically applying the hygienic or cosmetic compositions as described above in accordance with the utilization technique which is customary for these compositions. For example: application of creams, gels, solutions, lotions, makeup removing milks or sunscreen compositions or after-sun compositions to the dry skin or the dry hair, application of a hair lotion to wet hair, of shampoos, or else application of dentifrice to the gums.

Claim 1 of 19 Claims

What is claimed is:

1. An anti-inflammatory composition comprising an immixture of (1) at least one peptide derivative of .alpha.-type melanocyte stimulating hormone (.alpha.-MSH), and (2) at least one oligosaccharide solution obtained by enzymatic depolymerization of brown algal membrane polysaccharides.

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