NON-IRRITANT AQUEOUS LIPOSOME DISPERSIONS CONTAINING alpha -HYDROXYCARBOXYLIC ACIDS, alpha -KETOCARBOXYLIC ACIDS AND/OR SALICYCLIC ACID IN THEIR SALT FORMS

Abstract: The present invention relates to an aqueous liposomal dispersion of phospholipids comprising: (a) one or more alpha -hydroxycarboxylic acids, one or more alpha -ketocarboxylic acids, or salicyclic acid or mixtures therefrom and (b) one or more alcohols, said dispersion having a physiological pH value in the range from 5 to 7.5 and said acids being present nearly completely in salt form. The liposomal dispersion is nonirritant and has a beneficial effect on the skin.

Description: Non-irritant aqueous liposome dispersions containing α-hydro¬ xycarboxylic acids, α-ketocarboxylic acids and/or salicylic acid in their salt forms The present invention concerns non-irritant aqueous liposome dispersions of phosphol ipids for topical application, contain¬ ing α-hydroxycarboxylic acids, α-ketocarboxylic acids and/or salicylic acid in their salt forms. Diverse milk products and fruit ferments have been employed for centuries in traditional medicine and skin care, although at first it was not known which ingredients were responsible for their effects. Later, the α-hydroxycarboxylic acids or α-ketocarboxylic acids contained in these "beauty products" were found to have keratinolytic and skin stimulating proper¬ ties. Salicylic acid shows similar effects. For these reasons, individual members of these substance classes have been employed for some time in both dermatology and cosmetics. Recent investigations of the mechanism of action of the α-hydroxycarboxylic and α-ketocarboxylic acids have shown that their application to the skin corresponds to a gentle chemical peeling. A "softening" of the lipid material in the upper lay¬ ers of the stratum corneum accelerates the sloughing off of the outermost horny scales. In addition, the acids stimulate metabolism in the dermis, and natural skin functions are reac¬ tivated. The endogenous collagen and elastin synthesis activi¬ ties of the skin are stimulated. This, together with the accelerated scaling off of the outer skin layers, has a bene¬ ficial effect on the softness and elasticity of the skin. The skin becomes markedly softer and smoother. Clear improvements in greasy or unclean skin have also been observed through the use of α-hydroxycarboxylic and α-ketocarboxylic acids. Investigations have also shown that the effectiveness of the the substances decreases with increasing pH. According to test results presented on 21st March 1994 at the Advanced Techno¬ logy Conference in Barcelona, Spain, most of the α-hydroxycar¬ boxylic and α-ketocarboxylic acids showed very little efficacy at pH greater than 6. Optimal efficacy was observed at markedly lower pH values. However, the use of formulations of these acids or salicylic acid with a strongly acid pH leads to severe skin irritation. Thus while α-hydroxycarboxylic, α-ketocarboxylic acids and salicylic acid at low pH stimulate cell regeneration, they simultaneously cause irritation of the skin areas treated. Highly concentrated liposome dispersions containing malic acid, citric acid, glycolic acid or lactic acid for use in the manufacture of liposomal formulations are commercially avail¬ able (ROVI GmbH, Mutterstadt) . These preparations all have a pH value below 4.6, so as to ensure the effectiveness of the α-hydroxycarboxylic acid constituents. However, this low pH represents a considerable irritative potential when the dispersions are applied. EP-A-0 249 561, WO 94/12154, WO 93/15720, and WO 90/03781 des¬ cribe various pharmaceutical compositions containing liposomes and a α-hydroxycarboxylic acid or salicylic acid for different applications. EP-A-0 616 799 which is an earlier European patent application and does not belong to the state of the art discloses an aqueous cosmetic composition containing liposomes, un-neu- tralized free salicylic acid being included within the mem¬ brane lipid layer of the liposome, and an organic, water- soluble membrane-impermeant base. The object of the present invention is thus to provide a non- irritant formulation which contains α-hydroxycarboxylic acids, α-ketocarboxylic acids and/or salicylic acid in an effective form. This object is achieved by an aqueous liposomal dispersion of phosphol ipids comprising: (a) one or more α-hydroxycarboxylic acids, one or more α-ketocarboxylic acids, or salicylic acid or mixtures therefrom and (b) one or more alcohols, characterized in that said dispersion has a physiological pH value in the range from 5 to 7.5 and said acid(s) is (are) present nearly completely in salt form. Liposomes are spherical structures (vesicles) with an envelope of one or more lipid double layers (bilayers). They are usually manufactured from phosphol ipids and can be loaded with various active substances, which then become enclosed within the liposomes. The liposomes of the present invention have a preferred particle size of between 70 nm and 300 n (dia¬ meter) . One or more or a mixture of α-hydroxycarboxylic acids, α-keto¬ carboxylic acids and/or salicylic acid are enclosed in the vesicles of said aqueous liposome dispersion. In a preferred embodiment of the invention, liposomes from soya phospholipids with a phosphatidyl choline proportion of at least 73% by weight are used; a phosphatidyl choline pro¬ portion of between 73 and 79% by weight is more preferred. Phospholipid fractions can be used containing, for example, a) 73 to 79% by weight phosphatidyl choline, 0 to 6% by weight lysophosphatidyl choline, approx. 8% by weight phosphatidic acid, approx. 4% by weight phosphatidyl ethanolamine and approx. 9% by weight other lipids, or b) 90 to 96% by weight phosphatidyl choline and 0 to 6% by weight lysophosphatidyl choline or c) at least 95% by weight phosphatidyl choline and at most 2% by weight lysophosphatidyl choline. The fatty acid radicals of the soya phospholipids are mainly unsaturated fatty acids, linoleic acid being the major compo¬ nent. A typical fatty acid composition is: 61 to 71% linoleic acid, 3 to 7% linolenic acid, 6 to 13% oleic acid, 10 to 15% palmitic acid and 1.5 to 4% stearic acid. Alternatively, hydrogenated phospholipids with a phosphatidyl choline proportion of at least 73% by weight and preferably 75 to 95% by weight can be used. In principle, liposomes can be loaded with any dermatolo- gically active α-hydroxycarboxylic acid, α-ketocarboxylic acid or salicylic acid. Amongst the α-hydroxycarboxylic acids, gly¬ colic acid, lactic acid, citric acid, tartaric acid, malic acid, or mandelic acid are preferred. Pyruvic acid is the pre¬ ferred α-ketocarboxylic acid. The aqueous liposome dispersion contains one or more alcohols, preferably at a proportion of up to 20 % by weight. The alco¬ hol is preferably miscible with water and has a preserving effect. It advantageously has a vapour pressure being low enough to enhance penetration of the liposome dispersion into the skin. Monovalent alcohols are preferred. Suitable alcohols are for example ethanol and isopropanol, ethanol being more preferred. The aqueous liposome dispersion of the invention in a pre¬ ferred embodiment contains 5 to 20% by weight phospholipids, 5 to 30% by weight α-hydroxycarboxylic acids, α-ketocarboxylic acids or salicylic acid and no more than 20% by weight alco¬ hol. A more preferred composition contains 8 to 12% by weight phospholipids, 15 to 20% by weight acids and no more than 20% by weight alcohol. The said percent values for the acids refer to the total proportion of acids, irrespective of whether the acid molecules are dissociated or undissociated. The dissocia¬ ted acids are present in their salt form, usually as an alkali salt or alkaline earth salt, the sodium or calcium salts being preferred. The molecular weight of the undissociated acid molecule is used for the calculation of the proportion of acids by weight, irrespective of whether the acid molecules are actually dissociated or undissociated. In addition to the alcohol, the aqueous liposome dispersion can contain a preservative. Suitable preservatives are, for example, formaldehyde, parabene and Euxyl® K400. If the preservative is present together with alcohol, the alcohol proportion is then usually restricted to 1 to 2% by weight. The proportion by weight of water in the liposome dispersion is calculated from the proportions of the other components to 100%. The pH value of the liposome dispersion is in the range from 5 to 7.5. The ideal skin physiological pH is determined by the particular application. A pH in the range from 5.5 to 6.5 is preferred, since generally no skin irritation occurs at these pH values. For precise setting of the pH value, the aqueous liposome dispersion can contain a buffer system, such as phosphate buffer (KH P0 /Na HPO ). 2 4 2 4 The pH value set in the liposome dispersion and the acid con¬ stants of each α-hydroxycarboxylic and α-ketocarboxylic acid and salicylic acid determine to what extent the acid is pre¬ sent in dissociated form, i.e. as the salt. The pK values for a the α-hydroxycarboxylic acids given as examples above all lie within the range from 2.97 to 3.85, only the first dissocia¬ tion being considered in the case of multivalent carboxylic acids. From these values, using the law of mass action for equilibrium reactions, it can be calculated that the monova- lent acids are nearly completely dissociated at the pH value of the invention. For the multivalent acids, this means that at least one proton is dissociated. The aqueous liposome dispersion can be manufactured, for example, by first dissolving one or more α-hydroxycarboxylic and/or α-ketocarboxylic acids and/or salicylic acid and then adding a base until the required pH is reached. If the carbo¬ xylic acid salts are available, these can be dissolved in water and the solution brought to the desired pH by addition of an acid or a base. The salt solution thus obtained is mixed at high energy, for example with a high pressure ho ogenizer or a slit homogenizer, with a phospholipid mixture which has already been swollen in an aqueous medium to form a lamellar phase. The phospholipids spontaneously form a vesicular lamel¬ lar phase under mixing, which encloses the carboxylic acid salts. Further ho ogenization leads to a uniform distribution of the liposomes (polydispersity < 0.5). The dispersion can subsequently be diluted with alcohol and/or water. The aqueous liposome dispersions of the present invention have a beneficial effect on the skin, in that the α-hydroxycarboxy¬ lic and/or α-ketocarboxylic acids and/or salicylic acid pre¬ sent in their salt form show the keratinolytic and skin stimu¬ lating properties described above for the undissociated acids. In some cases, the 1 iposo e-enclosed salt forms of the acids have even been observed to be more effective than the undisso¬ ciated acids usually used. The liposome dispersions of the invention serve primarily as a base formulation for the manufacture of various liposome pre¬ parations for cosmetic and dermatological use. The preferred areas of application are regeneration and moisturization. The liposome dispersions improve the relief of the skin surface, the natural functions of the skin and sloughing, increase the moisture holding capacity and the elasticity of the skin, nor¬ malize keratinization and bring about a chemical peeling of the skin. The weak potency of the acids when not enclosed in liposomes, as in the current state of the art, at higher, non-irritant pH values, i.e. in the weakly acid or neutral range, can be taken to result from the fact that the acids in this pH range are nearly completely present in dissociated form as their salts. The acids in the liposome dispersion of the invention show surprisingly high potency, despite being present at physiolo¬ gical pH as their salts. The assumption that only the undisso¬ ciated form of the acid is effective must be questioned. Expe¬ riments which have been carried out, documented below with lactate as an example, clearly show that the liposome-enclosed acid residue anions possess a considerable skin stimulating effect. The "ineffectiveness" of the acid residue anions when not enclosed in liposomes would appear be due to the fact that the salt ions, in contrast to the undissociated acids, cannot penetrate the skin because of their electrical charge. Enclo- sure .of the salts in liposomes as a carrier system faciliates their penetration into the skin, where the acid residue anion, whose molecular structure is apparently responsible for the mechanism of action, is available to the skin. The following example of the manufacture of an aqueous liposo¬ me dispersion and the results of an experiment will serve to illustrate the present invention and to demonstrate the sur¬ prisingly high potency of the liposome dispersion on the skin. Example 1: Manufacture of an aqueous liposome dispersion con¬ taining lactic acid Composition: 29.3% by weight distilled water 20.0% by weight lactic acid 24.0% by weight sodium hydroxide solution (30%) 10.0% by weight phospholipid fraction, containing phosphatidyl choline (73-79% by weiσht) lysophosphatidyl choline ( 0-6 % by weight) phosphatidic acid ( < 8 % by weight) phosphatidyl ethanolamine ( < 4 % by weight) and other lipids (approx.9 % by weight) (proportions based on dry weights) 16.7% by weight ethanol, 96% Manufacture: Lactic acid is dissolved in a part of the distilled water and is carefully neutralized with sodium hydroxide solution to a pH of 6.8 to 6.9. The phospholipid fraction is swollen in the reamining distilled water and is added under homogenization using a homogenisator. Subsequently, ethanol is added under stirring. The result is a light yellow, moderately viscous, nearly transparent liposome dispersion. 8 Examp.le 2: Comparative study of the effect of lactate enclosed in liposomes in accordance with the present inven¬ tion, "free" lactate at pH 6.5 and "free" lactic acid at pH 1.8 with respect to skin moisture con¬ tent and surface structure. Three samples were tested in comparison with an untreated site (control) on a group of 5 female subjects aged between 28 and 50. Sample A: Aqueous liposome dispersion of the invention consisting of: 10% by weight phospholipids 20% by weight lactic acid 24% by weight sodium hydroxide solution (30%) 16% by weight ethanol 30% by weight distilled water pH 6.5 Sample B: Lactate solution consisting of: 20% by weight lactic acid 24% by weight sodium hydroxide solution (30%) 16% by weight ethanol 40% by weight distilled water pH 6.5 Sample C: Lactic acid solution consisting of: 20% by weight lactic acid 16% by weight ethanol 64% by weight distilled water pH 6 . 5 The measurements included changes in the moisture content and surface structure of the skin before and after 8, 21 and 28 days of twice daily application (morning and evening) by the experimental subjects. The inner surface of the forearm was selected as the area to be investigated, and the experimental sites were permanently marked. The measurements were carried out under controlled conditions in a biocl imatized room, a treatment-free period of at least 12 hours being allowed before each measurement (overnight, no morning application). The measurements of skin moisture content were carried out with the Corneometer CM 820 (Courage and Kazaka). This method is based on simple measurement of the capacitance of the stratum corneu . The underlying principle relies on the very different dielectric constants of water and of other substan¬ ces . Profilometry for the determination of skin surface structure was carried out with the analytical instrument Quantimet 970 together with a Polyvar Microscope from Leica/Cambridge Instruments, Bensheim. Video image data transfer from the skin surface takes place via an external macro apparatus. Contact- free imaging of the skin surface from various perspectives is carried out by means of a specially constructed apparatus, the combined data then being evaluated via image analysis. This technique is particularly well established for. the analysis of surface structures in the semiconductor industry and in mate¬ rial quality control. The principle variables of the comparative study were skin moisture content and the surface structure of the skin. The surface structure of the skin was recorded in terms of two measured parameters, skin smoothness and fold count. The variables were evaluated descriptively. The following Figures are based on the means of the measured values. Fig. 1: Skin moisture content Fig. 2: Profi lometry/skin smoothness Fig. 3: Profi lometry/fold count Evaluation of the results Skin moisture content: As shown in Fig. 1, the measurements with the corneometer revealed an increase in skin moisture content after only 8 days. No significant further change could be detected after 21 or 28 days. Sample A differed markedly from samples B and C in inducing a more marked increase in skin moisture content. The maximal effect on skin humidity of sample A after four weeks was 29%. Contact-free profilometry: The sites of measurement on the inner side of the forearm were assessed by means of a paired comparison, the same areas being compared before, during and after application. Using a 256- step gray scale, the size and depth of folds were evaluated and were used to calculate the skin smoothness and fold count in terms of % change in the skin surface. Both Fig. 2 and Fig. 3 show clear and contrasting effects of the samples on the surface structure of the skin. Sample A brought about a sub¬ stantial improvement in skin structure with respect to the depth and number of folds. Sample C also brought about an im¬ provement, although to a much lesser extent. Sample B actually led to a deterioration in skin structure. The experimental results clearly demonstrate the improved effectiveness of 1 iposo e-enclosed lactate compared with lac¬ tate not enclosed in liposomes and with lactic acid.

Claims: 11 C L A I M S 1. Aqueous liposomal dispersion of phospholipids comprising: (a) one or more α-hydroxycarboxylic acids, one or more α- ketocarboxyl ic acids, or salicylic acid or mixtures therefrom and (b) one or more alcohols, c h a r a c t e r i z e d i n t h a t said dispersion has a physiological pH value in the range from 5 to 7.5 and said acid(s) is (are) present nearly completely in salt form. 2. Aqueous liposomal dispersion of claim 1, c h a r a c t e r i z e d i n t h a t the pH value is in the range from 5.5 to 6.5. 3. Aqueous liposomal dispersion of claims 1 or 2, c h a r a c t e r z e d i n t h a t the proportion of said alcohol (s) is no more than 20 % by weight. 4. Aqueous liposomal dispersion of any of claims 1 to 3, c h a r a c t e r i z e d i n t h a t the proportion of said phospholipids is 5 to 20 % by weight, the proportion of said acid(s), on the basis of total dissociated und undissociated acid(s), is 5 to 30 % by weight, and the proportion of said alcohol (s) is no more than 20 % by weight. 5. Aqueous liposomal dispersion of any of claims 1 to 4, c h a r a c t e r i z e d i n t h a t said phospholipids are obtained from soya and comprise phosphatidyl choline at a proportion of at least 73% by weight. 6. Aqueous liposomal dispersion of any of claims 1 to 4, c h a r a c t e r i z e d i n t h a t said phospholipids are hydrogenated and comprise phospha¬ tidyl choline at a proportion of at least 73% by weight. 12 7. Aqueous liposomal dispersion of any of claims 1 to 6, c h a r a c t e r i z e d i n t h a t said α-hydroxycarboxylic acid is selected from glycolic acid, lactic acid, citric acid, tartaric acid, mal.ic acid, and mandelic acid. 8. Aqueous liposomal dispersion of any of claims 1 to 7, c h a r a c t e r i z e d i n t h a t said α-ketocarboxylic acid is pyruvic acid. 9. Aqueous liposomal dispersion of any of claims 1 to 8, c h a r a c t e r i z e d i n t h a t said alcohol is ethanol or isopropanol. 10. Aqueous liposomal dispersion of any of claims 1 to 9, c h a r a c t e r i z e d i n t h a t said liposomes have an average particle size of 70 to 300 nm.

Inventors: ROEDING JOACHIM, SEIGFRIED BERND G, TEICHMUELLER ERNST E, ROEDING, JOACHIM, SEIGFRIED, BERND, G., TEICHMUELLER, ERNST, E.
Applicants: ROEDING JOACHIM, ROVI GMBH, SEIGFRIED BERND G, TEICHMUELLER ERNST E, ROEDING, JOACHIM, SEIGFRIED, BERND, G., TEICHMUELLER, ERNST, E.
Application year: 1995
Main classification: A61K7
IPC codes: A61K8, A61K9, A61Q19