REACTION PRODUCTS OF HYALURONIC ACID AND NATURAL AMINO ACIDS AND THEIR USE IN COSMETIC AND PHARMACEUTICAL COMPOSITIONS

Abstract: The present invention relates to new high molecular weight salts of hyaluronic acid. More particularly, these salts are characterized in that they have as a counterion a natural amino acid. The products of the invention can be prepared starting from high molecular weight hyaluronic acid or salts thereof. These new high molecular weights salts of hyaluronic acid may be advantageously employed in the cosmetic of pharmaceutical field.

Description: REACTION PRODUCTS OF HYALURONIC ACID AND NATURAL AMINO ACIDS AND THEIR USE IN COSMETIC AND PHARMACEUTICAL COMPOSITIONS The present invention relates to new derivatives of hyaluronic acid, a process for their preparation and the cosmetic or pharmaceutical compositions containing such products. Hyaluronic acid is a mucopolysaccharide that is present almost in each part of a living organism and in particular in the skin. Chemically, hyaluronic acid is formed by straight polymer chains having a molecular weight ranging from hundreds thousand to millions dalton, including repeated disaccharid units constituted by N-acetyl glucosamine and glucoronic acid bound to each other by glucosidic bonds. Hyaluronic acid has the capacity of binding high amounts of water coming from the capillary network, necessary to keep in a solution form the catabolites, electrolytes and gases that spread from the capillaries to the cells and vice-versa, through the interstitial fluid. This phenomenon is of the essential to maintain the skin elastic and in a gelatinous physical condition. Hence, hyaluronic acid has a basic role in the control of the diffusion of nutrients, hormones, vitamins and inorganic salts of the connective tissue and in the removal of metabolism wastes which may give rise to reactions of an inflammatory type. As age increases, the amount of hyaluronic acid present in the organism decreases as well as its polymerization level, which results in a reduction in retained water, the skin shows alterations of its physical condition and undergoes an aging process that causes an increase in the fibrous part with respect to the elastic one. There are at present known cosmetic preparations that are mainly used as moisturizers or age retarding preparations, based on low molecular weight hyaluronic inorganic salts. Such preparations have the drawback of utilizing low molecular weight hyaluronic acid, which is much less effective compared to high molecular weight hyaluronic acid. On the other hand, high molecular weight hyaluronic acid is scarcely soluble, which creates severe technical problems in the preparation of cosmetic and pharmaceutical formulations. As a consequence, the need is still felt of providing derivatives of hyaluronic acid having a high effectiveness in the cosmetic and pharmaceutical field, and having also chemical-physical characteristics such as to allow their use in different types of formulations. The present invention relating to high molecular weight new derivatives of hyaluronic acid solves these problems. From one of his aspects, the present invention relates to new salts between high molecular weight hyaluronic acid and at least a natural amino acid. Preferred salts are those wherein the amino acid is selected from among lysine, arginine, methionine or aspartic acid. Particularly advantageous is lysine yaluronate which we gave the invention name Yalyse and which will be indicated in the future by the mark "YALYSE". By the expression "high molecular weight hyaluronic acid", there is intended to indicate a hyaluronic acid having a molecular weight comprised between 1.5 and 3 million dalton. The use of these new derivatives of hyaluronic acid in the cosmetic and pharmaceutical field provides remarkable advantages with respect to the known art. First of all, high molecular weight hyaluronic acid has the capacity of binding water amounts much higher than low molecular weight hyaluronic acid, promoting thereby a better diffusion of the dissolved substances and a greater elasticity of the skin. Besides this, the compounds of the present invention are constituted by hyaluronic acid salts with a counterion that can exercise a synergetic action with the yaluronate. In fact, as is known, amino acids carry out an essential role in the formation of special substances of skin physiology, such as melanin and keratin. Of particular importance is lysine, which is basic in the biosynthesis of melanin, as well as the collagen of elastin and reticulin which are the main proteic skin components. The derivatives of the present invention are therefore capable of markedly reducing skin aging through an increase moisturizing of the dermis, a promoted cellular recycling and a stimulation of the synthesis of elastin as well as of free radicals. From another of its aspects, the present invention relates to a process for the preparation of salts selected from among hyaluronic acid and at least a natural amino acid, characterized in that hyaluronic acid or a salt thereof is caused to react with at least a natural amino acid, in the form of a free base or in a salified form, respectively. According to an advantageous aspect, the present invention relates to a process for the preparation of lysine hyaluronate, characterized in that sodium hyaluronate is reacted with lysine hydrochloride. Preferably, such reaction is carried out by suspending sodium hyaluronate in water and stirring until it gels before adding lysine hydrochloride. The preparation reactions of the salts subject matter of the present patent application may be carried out using a stochiometric amount of the reagents or an excess of either component, in which latter case one obtains the salt and an excess reagent co-ordination through hydrogen bonds. By the expression "stochiometric amounts of the reagents" there is intended to indicate that the number of amino acid moles used is equal to the number of the carboxy functions that are present in hyaluronic acid. The possibility of varying the ratio of acid moles to amino acid moles allows to modulate the characteristics of the products obtainable as a function of the desired properties. Organic molecules of various type may be coordinated to the products obtained according to the aforesaid processes by means of hydrogen bonds. This allows to obtain end products having variable chemical-physical and pharmacological properties. From another of its aspects, the present invention relates to the use of new high molecular weight hyaluronic acid salts in the cosmetic field. From a further aspects the present invention relates to the use of the new high molecular weight hyaluronic acid salts as medicaments. The active principles of the present invention are adapt to be formulated with suitable excipients, for instance those usually utilized is the cosmetic field, such as bee-wax, jujube oil, isostearyl isostearate, fatty acid, triglycerides, propylen glycol, lauryl alcohol, and hydroxypropyl methyl cellulose. Stabilizers, antioxidants, and preservatives, either individually or combined with each other, may be added to these active principles. Suitable stabilizers include Acepur K, Abiol, Prevan, Fondix G, BHA, BHT. Suitable antioxidants include butylhydroxyanisole, citric acid, tocopherol, sodium thiophosphate. Suitable preservatives include p-hydroxybenzoic acid esters or imidazolidinyl urea. A preferred composition of the invention is in the one that comes in the form of an emulsion. Emulsions containing 1 % to 5% lysine hyaluronate (Yalyse) are particularly preferred. Such compositions, either in the pure form or in the form of lyophilized sheets, are used in the treatment of burn wounds, decubitus ulcers, venous ulcers, and in all cicatrization processes. They may be also advantageously used in aesthetic surgery, especially following epidermis abrasion laser therapy interventions, in order to achieve a quicker and optimum re-epithelization. Another preferred composition of the present invention in the one that comes in the form of a lotion. Particularly advantageous are the lotions comprising 1 % to 7% arginine hyaluronate, possibly mixed with glycolic acid and carrier substances and suitable excipients compatible with the type of utilization desired. Such lotions may be employed in alopecia cases due to excess sebum production, in allergy-based scalp hyperkeratoses, and in dandruff persistent cases. A further type of preferred composition of the present invention is the one that comes in the form of a cream. Particularly advantageous are creams based on 1 % to 7% lysine hyaluronate (Yalyse), possibly mixed with one or more substances selected from among vitamin A, vitamin E, ascorbic acid, cogic acid, allantoin, echinaceous calendula, "centella", horsetail, persea, propolis, aloe, cera alba, arachis, zea mays, horse chestnut, ginko biloba, jujube, shea butter, bee-wax, hazelnut, sunflower, and with excipients compatible with its specific utilization. Thanks to their moisturizing action, such creams are particularly useful in reducing the process of skin-aging, in the treatment of wrinkles, dryness, opacity, and in promoting skin elasticity and luminosity. These creams may also be used as protectors in photodermatosis, as anti-erythema and, together with cogic acid or hydroquinone, in the treatment of senile spots. From a last aspect, the invention relates to the use of new high molecular weight hyaluronic acid salts for the preparation of medicaments suitable for the treatment or prevention of a pathologic condition selected from among psoriasis, eczema, ichthyiosis, postactinic radiodermatosis, hemorrhoids, proctitis, hemorrhagic proctitis, atopic dermatitis, trophic ulcers and decubitus ulcers, sudamina, epidermophytosis or, generally, in the cases of prurigo. In these cases, the pharmaceutical compositions may possibly also include one or more substances selected from among salicylic acid, ascorbic acid or retynyl palmitate. In the following, some examples of embodiments of the present invention are reported, only by way of non-limiting example. EXPERIMENTAL PART Process for the preparation of lysine hyaluronate 1 g sodium hyaluronate (corresponding to 0.945 g hyaluronic acid) is reacted with 0.455 g lysine hydrochloride (0.364 g free base lysine), obtaining 1.31 g lysine hyaluronate and 0.145 g NaCI. The preparation of the product takes place according to the following steps: a) preparation of gel by mixing sodium hyaluronate with water; b) preparation of lysine hydrochloride; c) addition of lysine hydrochloride to sodium hyaluronate gel; d) freeze-drying of the product. Gelation of sodium hyaluronate 10 g sodium hyaluronate are suspended under stirring in 300 ml of deionized water, up to full gelation. The product is clear, colorless, homogeneous and free from clots. The gel obtained has a pH of 6.6. The hyaluronic acid/water ratios are indicative; greater water amounts may be used (either at the start or upon completion of the product), should one wish not to proceed to drying or freeze-drying the product, utilizing instead directly the gel, in order to obtain a product having the most suitable physical properties (for instance, viscosity). Preparation of lysine hydrochloride 7.3 g 50% free base lysine are diluted in 20 ml water (corresponding to 3.65 g lysine), and 25 ml HCL 1 N (corresponding to 0.91 g HCI) are added under stirring. Before the addition of the acid, the solution has a pH of 9.9, upon completion of the acidification, the pH is of 5.7. Synthesis of lysine hyaluronate Lysine hydrochloride is added to the previously prepared sodium hyaluronate gel, slowly and under stirring; once the addition is completed, stirring is maintained for about 1 hour, to obtain the complete homogenization of the product. The product has a pH of 6.5. Should one wish to modify the pH of the product, an acid or a base is added under the control of a pH-meter, until the pH desired is obtained. Other organic molecules may be added to the product obtained, which are co-ordinated by means of hydrogen bonds. The product is allowed to rest overnight in a freezer, and the following day it is freeze-dried or vacuum dried. Following the same process, it is possible to prepare: arginine hyaluronate, methionine hyaluronate, and aspartic acid hyaluronate. The synthesis diagrams for the reactions carried out starting from non- salified products are shown hereunder. LYSINE HYALURONATE n ARGININE HYALURONATE H 2 METHIONINE HYALURONATE OOH SUBSTΓΓUTE SHEET (RULE 26) ASPARTIC ACID HYALURONATE COOH Preparation of the scalp lotion The preserving substances are vacuum loaded in a turboemulsifier at a temperature of 80°C. Afterwards, the remaining matters are introduced under stirring one by one, except for arginine hyaluronate. After 15' mixing at a constant speed, the temperature is cooled to 40°C, arginine hyaluronate is added and mixing is continued until the temperature of 25°C is reached. The obtained mixture is filtered with a polycarbonate filter with 0.45 μ holes. The end formulation has the following composition: DEIONIZED WATER Q.S. 100 GLYCOLIC ACID % 5 ARGININIE HYALURONATE % 3 UNDECYLENAMIDE DEA % 1 PG-HYDROXYETHYL CELLULOSE % 0.9 COCODINIUM CHLORIDE % 0.75 SODIUM HYDROXIDE % 0.75 POLYQUATERNARIUM-10 % 0.6 ZINC P.C.A. % 0.3 IMIDAZOLIDINYL UREA % 0.3 BENZOPHENONE % 0.05 Preparation of age-retarding cream All the excipients are vacuum loaded in two turboemulsifiers at a temperature of 80°C. Emulsifier-A) - all water-soluble excipients. Emulsifier-B) - all oily excipients. Except for: lysine hyaluronate, elastin, collagen, tocopherol and fragrance. Both meltings are stirred until the temperature reaches 80°C. Now both meltings are united and mixed for 15' at a constant speed, lowering temperature to 40°C. Now the matters left by side are added, and stirring is continued until the temperature reaches 25°C. The end formulation has the following composition: DEIONIZED WATER Q.S. 100 PROPYLEN GLYCOL 5 GLYCERYL STEARATE % 5 OCTYL STEARATE 3.5 CETEARYL ALCOHOL % 3.5 LYSINE HYALURONATE 3 STEARETH-30 % 2.7 CETETH-2 2.7 SHEA BUTYRUM % 2.4 ELASTIN % 1 SOLUBLE COLLAGEN 1.5 OLEA EUROPEA CALENDULA OFFICINALIS 1 PERSEA GRATISSIMA % 1 CERA ALBA 1 TRIISOSTEARIN % 1 TOCOPHEROL (Vit. E) 1 RETYNYL PALMITATE (Vit. A) 0.3 PHENOXYETHANOL 0.3 ASCORBIC ACID (Vit. C) 0.2 FRAGRANCE 0.2 BHA 0.1 POLYSORBATE 20 0.1 DICHLOROBENZYL ALCOHOL 0.1 DEHYDROACETIC ACID 0.1 TRIETHANOLAMINE 0.1 SODIUM METHYL PARABEN 0.09 SODIUM PROPYL PARABEN 0.03 ARACHIS HYPOGAEA 0.01 BHT 0.0003 PROPYL GALLATE 0.0003 CITRIC ACID 0.0001 PREPARATION OF THE EMULSION All the excipients are vacuum loaded in a turboemulsifier at a temperature of 80°C, except for lysine hyalurate. The whole is turbo-stirred for 15', at a constant speed. Afterwards, always under stirring, the temperature is cooled to 40°C. When this temperature has been reached, lysine hyalurate is added and mixing is continued slowly until the emulsion is cooled and a temperature of 28°C is reached. The obtained mixture is then submitted to bacteriologic control. The end formulation has the following composition: DEIONIZED WATER Q.S. 100 ARGININIE HYALURONATE % 3 PROPYLEN GLYCOL % 1.3 RETYNYL PALMITATE (Vit. A)) % 0.3 IMIDAZOLIDINYL UREA % 0.3 ASCORBIC ACID (Vit. C) % 0.2 SODIUM METHYL PARABEN % 0.15 SODIUM PROPYL PARABEN % 0.09

Claims: CLAIMS 1. Products of salification between hyaluronic acid and at least a natural amino acid. 2. The products according to claim 1 , characterized in that said hyaluronic acid is a high molecular weight hyaluronic acid. 3. The products according to claim 1 , characterized in that said amino acid is selected from among lysine, arginine, methionine or aspartic acid. 4. Lysine hyaluronate. 5. Process for the preparation of hyaluronic acid salts according to claim 1 , characterized in that hyaluronic acid or a salt thereof is reacted with at least a free or a salified amino acid. 6. Process for the preparation of lysine hyaluronate, characterized in that the reaction is carried out starting from sodium hyaluronate and lysine hydrochloride. 7. The process according to claim 6, characterized in that sodium hyaluronate is suspended in water and stirred to gelification before the addition of lysine hydrochloride. 8. Use of the compounds of claim 1 in the cosmetic field. 9. Use of the compounds of claim 1 as medications. 10. Cosmetic or pharmaceutical compositions comprising the compounds of claim 1. 11. The compositions according to claim 10, characterized in that they come in the form of an emulsion. 12. The compositions according to claim 11 , characterized in that they comprise 1 % to 5% lysine hyaluronate. 13. The compositions according to claim 12, characterized in that they also comprise deionized water, propylen glycol, retynyl palmitate, imidazolydinyl urea, ascorbic acid, sodium methylparaben and sodium propylparaben. 14. The use of the compositions according to claims 11-13 for the treatment of decubitus ulcers, venous ulcers or cicatrization processes. 15. The compositions according to claim 10, characterized in that they come in the form of a lotion. 16. The compositions according to claim 15, characterized in that they comprise 1 % to 7% argine hyaluronate. 17. The compositions according to claim 16, characterized in that they also comprise deionized water, glycolic acid, undecylen urea, PG-hydroxyethyl cellulose, cocodinium chloride, sodium hydroxide, polyquaternarium-10, p.c.a. zinc, imidazolydinyl urea and benzophenone. 18. The use of the compositions according to claims 15-17, for the treatment of alopecia, scalp hyperkeratoses, and dandruff. 19. The compositions according to claim 10, characterized in that they come in the form of cream. 20. The compositions according to claim 19, characterized in that they comprise lysine hyaluronate. 21. The compositions according to claim 20, characterized in that they also comprise: DEIONIZED WATER Q.S. 100 PROPYLEN GLYCOL 5 GLYCERYL STEARATE % 5 OCTYL STEARATE 3.5 CETEARYL ALCOHOL % 3.5 LYSINE HYALURONATE 3 STEARETH-30 % 2.7 CETERH-2 2.7 SHEA BUTYRUM % 2.4 ELASTIN % SOLUBLE COLLAGEN OLEA EUROPEA CALENDULA OFFICINALIS PERSEA GRATISSIMA % CERA ALBA TRIISOSTEARIN % TOCOPHEROL (Vit. E) RETYNYL PALMITATE (Vit. A) 0.3 PHENOXYETHANOL 0.3 ASCORBIC ACID (Vit. C) 0.2 FRAGRANCE 0.2 BHA 0.1 POLYSORBATE 20 0.1 DICHLOROBENZYL ALCOHOL 0.1 DEHYDROACETIC ACID 0.1 TRIETHANOLAMINE 0.1 SODIUM METHYL PARABEN 0.09 SODIUM PROPYL PARABEN 0.03 ARACHIS HYPOGAEA 0.01 BHT 0.0003 PROPYL GALLATE 0.0003 CITRIC ACID 0.0001 22. The use of the compositions according to claims 19-21 , for the treatment of wrinkles, skin dryness, opacity and aging processes. 23. The use of the compounds according to claim 1 in the preparation of medicaments suitable for the prevention or the treatment of pathologic conditions such as psoriasis, eczema, ichthyiosis, postactinic radiodermatosis, hemorrhoids, proctitis, hemorrhagic proctitis, atopic dermatitis, trophic ulcers and decubitus ulcers, sudamina, epidermophytosis or, generally, in the cases of prurigo. 24. Compositions comprising the compounds of claim 1 together with one or more substances selected from salicilic acid, ascorbic acid or retynyl palmitate.

Inventors: ABBIATI GIULIANA, ABBIATI, GIULIANA
Applicants: ABBIATI GIULIANA, JASPER LTD LIABILITY CO, ABBIATI, GIULIANA, JASPER LTD. LIABILITY CO.
Application year: 1999
Main classification: C08B37
IPC codes: A61K8, A61K9, A61K31, A61K47, A61P1, A61P9, A61P17, A61P35, A61Q5, A61Q7, A61Q9, A61Q19, C08B37