Composition for treating the skin of diabetic patients

Abstract

1. A pharmaceutical or cosmetic composition comprising an active amount of divalent calcium for use in the topical treatment of skin having compromised barrier function, particularly preferably the skin of diabetic patients, wherein the divalent calcium is present in the composition as a phospholipid complex.

Description

[Technical field]

[0001] The present invention relates to a composition for treating the skin of diabetic or pre-diabetic patients, in particular for preventing dry skin or for restoring well-hydrated skin and / or for restoring the barrier effect and / or for preventing wrinkles. Furthermore, the composition preferably brings about a reduction in itching, redness, blistering, furunculosis and / or tightness. The present invention further relates to the use of such a composition for therapeutic or cosmetic purposes, and to a method for preparing such a composition. [Background technology]

[0002] Skin changes occur in the majority of people with diabetes, not only in the later stages of the disease, but also as early as the pre-diabetic stage, when blood sugar levels are high. Pathological skin changes are promoted by poor metabolic control. Skin disorders, such as especially dry skin, with its associated itching, blistering, redness, wrinkles, or furuncles, can be due to poor control of the diabetic disease, but even with good control, it is often impossible to prevent especially dry skin. Therefore, diabetic patients should use special products to care for their sensitive skin. Even if the metabolism is well controlled, people with diabetes are more easily susceptible to fungal infections and other skin infections, and about one-third of people suffer from skin dysfunction due to excessively high blood sugar levels. The skin of diabetic patients is similar to aged skin. In particular, the barrier function of the skin is impaired.

[0003] For the care of dry skin in diabetics, cosmetics or medicines that provide good skin hydration are available, but these products only control the symptoms of the skin in diabetics, not its cause.

[0004] Good skin hydration is achieved by using occlusive lipids such as shea butter, lanolin, mineral oil, and many other fats. In addition, these products use high concentrations of moisturizing elements such as urea. As a result of all these components, these care products are not very pleasant to handle. They are sticky and transfer quickly to clothes.

[0005] Devaz and Pal, in J. Clin. Physiol. 1999, 143:131-132, describe the preparation of the helices as phospholipid-calcium precipitates with calcium to lecithin molar ratios in the range of 1:2. No therapeutic use is indicated.

[0006] Hirotsuka et al., in Non-Patent Document 2, describe how calcium ions can be provided in the form of lecithin liposomes, but again no therapeutic applications are indicated. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] “Abstracts for 6th Central European Symposium on Pharmaceutical Technology and Biotechnology ED - Zakelj Simon; Mrhar Ales; Gra”, EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES, ELSEVIER AMSTERDAM, NL, vol. 25, 1 May 2005 (2005-05-01), P-27, page S83 - page S84 [Non-Patent Document 2] “Calcium fortification of soy milk with calcium-lecithin liposome system”, JOURNAL OF FOOD SCIENCE, WILEY-BLACKWELL PUBLISHING, INC, US, vol. 49, no. 4

[0008] Disclosure of the Invention It is therefore, inter alia, an object of the present invention to propose a pharmaceutical or cosmetic composition for topical application, i.e. in particular as a cream or ointment, which provides relief or even improvement of skin properties to persons with skin with compromised barrier function and / or to diabetic or pre-diabetic patients, in particular to patients with skin that does have compromised barrier function.

[0009] In the present specification, skin with impaired barrier function should be understood to mean skin that exhibits increased transepidermal water loss (TEWL). This variable is measured, for example, with the aid of the Tewameter® TM300 instrument from Courage+Khazaka, Germany. Thus, skin with impaired barrier function typically exhibits a TEWL value of 12 g / m 2 / h, preferably 15 g / m 2 / h, particularly preferably 20 g / m 2 / h or even higher than 25 g / m 2 / h. Conventionally, this value is measured on the inside of the forearm.

[0010] In other words, skin with impaired barrier function typically has TEWL values ​​at least 4 g / m below normal. 2 / h, preferably at least 7 g / m 2 / h, particularly preferably at least 15 g / m 2 / h, or even at least 20 g / m 2 / h (normal measurements are also assumed to be on the inside of the forearm. Healthy adult subjects typically have a skin rash of 6.8 g / m2, also measured on the inside of the forearm. 2 / h (see, e.g., Akdeniz, M., Gabriel, S., Lichterfeld-Kottner, A., Blume-Peytavi, U. and Kottner, J. (2018), TEWL reference values ​​in healthy adults. Br J Dermatol, 179: e204-e204. https: / / doi.org / 10.1111 / bjd.17215).

[0011] Such pharmaceutical or cosmetic compositions for topical application are therefore proposed in particular for the prevention of dry skin or for the restoration of well-hydrated skin and / or for the restoration of the barrier effect and / or for the prevention of wrinkles. Furthermore, the compositions preferably bring about a reduction in itching, redness, blistering, furuncles and / or tightness. In particular, it is intended that not only the symptoms of dry skin but also its causes are controlled. Furthermore, it is intended that the convenience of application is improved.

[0012] It is intended that restoration of the skin barrier and thus skin hydration is particularly preferably achieved without occlusive fats and urea.

[0013] Thus, the proposed formulations as semi-processed products, but in particular whose dosage forms are intended for use as, for example, ointments, creams, lotions, pastes or tinctures for topical application to the patient's skin, are preferably urea-free (or contain less than 1% by weight, or less than 0.5% by weight, or less than 0.25% by weight of urea) and / or are formulated so as not to form an occlusive layer in the form of a film on the skin (are non-occlusive). The proposed formulations are therefore particularly preferably free of (or contain less than 1% by weight, or less than 0.5% by weight, or less than 0.25% by weight of) siloxanes, petrolatum, lanolin, mineral oil and / or other occlusive substances or combinations thereof, in particular those selected from the group of petrolatum, C18-C30 alkylmethylsiloxanes, dimethicone, polymethylsilsesquioxane, lanolin or lanolin alcohol, mineral oil (liquid paraffinum), which substances are preferably absent or present in amounts less than 1% by weight, or less than 0.5% by weight, or less than 0.25% by weight, whether in combination or individually.

[0014] The present invention therefore provides a pharmaceutical and / or cosmetic composition according to claim 1 or the use of such a composition for therapeutic and / or cosmetic treatment and a method for therapeutic and / or cosmetic treatment, as well as a method for preparing such a composition.

[0015] According to a first aspect, the present invention relates to a pharmaceutical or cosmetic composition having an active amount of divalent calcium for use in the topical treatment of the skin in diabetic or pre-diabetic patients, wherein the divalent calcium is present in the composition as a phospholipid complex.

[0016] Preferably, the phospholipid complex is in the form of an aqueous gel in such compositions and / or in the starting material containing calcium phospholipids of the composition. Gel should be understood to mean a non-fluid colloidal or polymeric network whose total volume is expanded by a fluid, according to the general knowledge in the art, in line with the IUPAC Gold Book (https: / / doi.org / 10.1351 / goldbook.G02600). Gels have a finite, usually rather small, yield stress. Gels may contain covalent polymeric networks, for example networks formed by crosslinked polymer chains or non-linear polymerization. Gels may also contain polymeric networks formed by physical aggregation of polymeric chains by hydrogen bonding, crystallization, helical formation, complexation, etc., resulting in locally ordered regions that are the junctions of the network. The resulting swollen network can be called a thermoreversible gel if the locally ordered regions are thermoreversible. Gels may also contain polymeric networks formed by glassy junctions, for example those based on block copolymers. If the junctions are thermoreversible glassy domains, the resulting swollen network can also be called a thermoreversible gel.

[0017] It has been surprisingly found that the formulation of divalent calcium as a phospholipid complex, especially a special phospholipid complex having a biconical structure, is much more accessible for topical application, for example as a concentrate and also as a cream or ointment, than, for example, when calcium is added to such formulations as a simple chloride or similar salt, in particular to restore the skin barrier provided by an intact stratum corneum.

[0018] The phospholipids present in the proposed complexes are not effective when applied topically alone, for example as a phospholipid mixture, as explained below.It is the formulations containing phospholipid complexes in specific combinations with calcium that restore the skin barrier.

[0019] As explained below, this form of divalent calcium preparation can stabilize, reduce or even reverse typical skin problems in diabetic patients. These include, in particular, the treatment of skin problems due to diabetes or prediabetes, in particular dry skin or moisturizing dry skin, atopic skin conditions, restoring the barrier function of the skin, skin infections, in particular of bacterial origin, or fungal infections due to a lack of barrier function, pigmentation disorders (diabetic dermopathy), blistering, itching (diabetic pruritus), redness (including diabetic necrobiosis lipoidica, pseudoacanthosis nigricans, diabetic blistering, diabetic rubeosis, diabetic scleredema), wrinkles, skin scaling, tightness, impaired wound healing, elasticity.

[0020] Preferably, the proposed formulation is used for the dermatological treatment of diabetic patients, preferably type 2 patients.

[0021] The proposed composition is preferably characterized in that the phospholipid is a phosphoglyceride (lecithin) selected from the group of phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, diphosphatidylglycerol (cardiolipin) or mixtures of these systems, which may be totally or partially hydrogenated or not.

[0022] It is particularly preferred that the calcium phospholipid complexes in the composition are, at least in part, calcium complexes with negatively charged phospholipids, particularly phosphatidic acid, phosphatidylinositol, phosphatidylserine, and the like.

[0023] In this case, the molar ratio of divalent calcium to phospholipid (based on the total amount of phospholipid, i.e., negatively charged phospholipid and uncharged phospholipid) is preferably in the range of 0.05:1 to 20:1 (i.e., 0.05 to 20), preferably in the range of 0.1:1 to 5:1 (i.e., 0.1 to 5), particularly in the range of 0.2:1 to 1:1 (i.e., 0.2 to 1), preferably in the range of 0.3:1 to 0.8:1 (i.e., 0.3 to 0.8).

[0024] In particular, the system is characterized in that the calcium phospholipid complexes are preferably at least partially neutral complexes in the form of biconical structures and are therefore easily bioavailable (according to FIG. 1b). These structures are characterized in that the phospholipids are 2+ When properly mixed with a solution, it is formed and in an aqueous environment, 2+ A cigar-shaped structure is formed that stabilizes the bicone (Fig. 1a). The complex is particularly 2+ However, phospholipids can also be partially charged and uncharged, i.e., Ca2+, Ca2++, Ca2++, Ca2++, and Ca2++. 2+ Since the phospholipids can form complexes with the phospholipids, it is possible for the complexes to be formed wholly or partially with uncharged or charged phospholipids, or partially with uncharged phospholipids and partially with charged phospholipids in the composition.

[0025] Thus, according to a further preferred embodiment, the calcium phospholipid complex is a calcium complex with phospholipids, in particular at least partially with phosphatidic acid, phosphatidylinositol and / or phosphatidylserine, in which the molar ratio of divalent calcium to negatively charged phospholipid, preferably phosphatidic acid, phosphatidylinositol and / or phosphatidylserine, based on the content of negatively charged phospholipid, is in the range of 0.1:1 to 30:1 (i.e. 0.1 to 30), preferably in the range of 0.3:1 to 10:1 (i.e. 0.3 to 10), more preferably in the range of 0.4:1 to 5:1 (i.e. 0.4 to 5) or in the range of 0.5:1 to 1:1 (i.e. 0.5 to 1).

[0026] Ca 2+Phospholipid complexes can be prepared by using conventional lecithins derived from soybean, egg, sunflower, and other sources. These lecithins have different phospholipid compositions. They have different head groups and different fatty acid compositions. Furthermore, these phospholipids in lecithins can be further processed to obtain new properties, for example, by hydrogenating the fatty acids to increase stability, or by separating the head groups or fatty acids from the glycerol structure to make the phospholipids smaller. In addition to natural sources, pure phospholipids from synthesis or purification can also be used. Furthermore, phospholipid-like molecules with negatively charged polar head groups and hydrophobic tails can also be used. These molecules can be found in the field of emulsifiers and wash-active materials.

[0027] In the composition, for the proposed use, Ca is intended for topical application. 2+ The concentration of divalent calcium in the phospholipid complex is typically in the range of 0.001 to 10 weight percent, preferably in the range of 0.005 to 5.0 or 0.01 to 0.5 weight percent (weight percent of CaCl2·2H2O based on the total composition).

[0028] In the composition, for the proposed use, Ca is intended for topical application. 2+ The concentration of phospholipid in the phospholipid complex is typically in the range of 0.01 to 30 weight percent, preferably in the range of 0.05 to 5 weight percent (weight percent of phospholipid based on total composition).

[0029] The proposed compositions can be formulated as ointments, creams, lotions, pastes, or tinctures for topical application to the patient's skin.

[0030] The composition then preferably additionally contains at least one of the formulation components thickeners, lubricants, moisturizers and / or humectants, surfactants, preservatives, antifoamers, waxes, fats, oils, antioxidants or substances with antioxidant properties, bactericides, fungicides, fragrances, propellants, dyes, stabilizers, polar and non-polar solvents, in particular water, pigments, UV filters, plant extracts, further active therapeutic or medicinal ingredients, or combinations thereof.

[0031] The composition may also be in the form of a concentrate, e.g., a bulk material for the preparation of end-use formulations, and for the preparation of topical formulations, the composition may contain Ca. 2+ The concentration of the phospholipid complex is in the range of 0.1 to 100 weight percent, preferably in the range of 1 to 10 weight percent (Ca based on the topical composition). 2+ The weight percent of the phospholipid complex may be designed.

[0032] The present invention further relates to the use of such compositions for the treatment and / or cosmetic treatment of the skin of diabetic patients or to a method for the treatment and / or cosmetic treatment of the skin of diabetic patients. In particular, the present invention relates to the stabilization, prevention, reduction or elimination of dry skin or moisturizing dry skin and / or restoring impaired barrier function of the skin. Alternatively or additionally, the present invention relates to the stabilization, prevention, reduction or elimination of skin infections, in particular of bacterial origin, due to impaired barrier function, skin changes or skin irritation, or the stabilization, prevention, reduction or elimination of fungal infections, pigmentation disorders (e.g. diabetic dermopathy), blistering, itching (e.g. diabetic pruritus), redness (including diabetic necrobiosis lipoidica, pseudoacanthosis nigricans, diabetic blisters, diabetic rubeosis, diabetic scleredema), wrinkles, or else furunculosis and / or tightness, wherein the above compositions are topically applied to the skin.

[0033] The compositions are preferably used in a form formulated as an ointment, cream, lotion, paste, or tincture for topical application to the patient's skin.

[0034] The present invention further relates to a method for preparing the above-mentioned composition, characterized in that the phospholipid starting material, optionally after having been previously dissolved or dispersed in an organic solvent, in particular ethanol, or a mixture of an organic solvent and water, is provided in an aqueous medium as liposomal structures, preferably having an average particle size of less than 300 nm, preferably less than 200 nm.

[0035] This dispersion was then 2+ The aqueous solution is preferably adjusted to a pH range of 7.5 to 10, and more preferably to a pH range of 8 to 9.

[0036] The liposome-containing dispersion can be added to the aqueous calcium solution, or the liposome-containing dispersion can be first charged and then the calcium solution added. This is preferably done with stirring. Once this process is complete, the Ca 2+ is in the form of a bicone at the appropriate concentration, temperature, and phospholipid to calcium ratio.

[0037] These steps are preferably followed by homogenization to form a gel and thus the aggregates described.

[0038] Preferably, the preparation and / or homogenization of the liposomes proceeds using a high-pressure homogenizer, preferably at a pressure of at least 500 bar, particularly preferably at least 1000 bar.

[0039] The concentrated composition can then be formulated into an ointment, cream, lotion, paste, or tincture using at least one carrier substance and optionally further components.

[0040] In other words, Ca 2+ The phospholipid complexes (particularly bicones or aggregates thereof) can be prepared as follows.

[0041] Phospholipid molecules, such as lecithin, are transferred to the aqueous phase, which may additionally contain further components, such as ethanol, glycerol, panthenol, propylene glycol, further active ingredients such as moisturizers and active anti-aging ingredients, preservatives, dyes, odor substances, stabilizers, and other cosmetic / medical ingredients. Phospholipids can be transferred to the aqueous phase by swelling or by pre-dissolving in a solvent such as ethanol. Liposome structures of phospholipids (lecithin) are preferably prepared in the aqueous phase. Known methods of liposome preparation can be used, such as high pressure homogenization, extrusion, dialysis, ultrasound, and further methods.

[0042] Next, Ca 2+ A solution (obtained, for example, by dissolving calcium chloride CaCl2·2H2O) is slowly added to the phospholipid (lecithin) phase while stirring. 2+ In this mixture, the desired composite structures, especially biconical Ca 2+ The correct concentration must be chosen so that the structure (Figure 1b) can be formed. The correct ratio depends on the phospholipid (lecithin) used and can be tested in each individual case. Depending on the total concentration of phospholipids, the Ca 2+ The formation of phospholipid bicone complexes takes on a gelatinous consistency and transitions into aggregates (Figure 1a).

[0043] Further embodiments are defined in the dependent claims.

[0044] Preferred embodiments of the present invention are described below based on the drawings, which are merely used for illustration purposes and should not be construed as limiting. [Brief description of the drawings]

[0045] [Figure 1a] FIG. 1 shows a complex phospholipid-Ca2+ structure, showing aggregates of biconical structures. [Figure 1b] FIG. 1 shows complex phospholipid-Ca2+ structures, showing elementary and individually occurring biconical structures. [Figure 2a] Hematoxylin-eosin staining of human 3D epidermis after 9 days of differentiation treated with different calcium concentrations or Ca2+phospholipid complexes, showing control (1.1 mM CaCl2·2H2O on the basal side). [Figure 2b] Hematoxylin-eosin staining of human 3D epidermis after 9 days of differentiation treated with different calcium concentrations or Ca2+phospholipid complexes shows reduced calcium concentration (basolateral 0.3 mM CaCl2·2H2O) and impaired epidermis formation was observed at reduced calcium concentration (b). [Figure 2c] Hematoxylin-eosin staining of human 3D epidermis after 9 days of differentiation treated with different calcium concentrations or Ca2+phospholipid complexes shows treatment with CaCl2·2H2O (0.3 mM CaCl2·2H2O on the basal side and 1.1 mM CaCl2·2H2O on the apical side) and impaired epidermis formation was observed with treatment with 1.1 mM calcium chloride dihydrate solution on the apical side (c). [Figure 2d] Hematoxylin-eosin staining of human 3D epidermis after 9 days of differentiation treated with different calcium concentrations or Ca2+ phospholipid complexes shows that treatment with Ca2+ phospholipid complexes (basolateral 0.3 mM CaCl2·2H2O and apical 0.1% Ca2+ phospholipid complexes, preparation similar to sample 6 in Table 1) supports the formation of a normal epidermis. [Figure 3a] Figure showing loricrin expression staining of 3D epidermis after 9 days of differentiation treated with different calcium concentrations or Ca2+phospholipid complexes, showing control (1.1 mM CaCl2·2H2O on the basal side). [Figure 3b] Figure 14. Loricrin expression staining of 3D epidermis after 9 days of differentiation treated with different calcium concentrations or Ca2+phospholipid complexes. Reduced calcium concentration (basolateral 0.3 mM CaCl2·2H2O) is shown. Decreased loricrin production was observed at reduced calcium concentration (b). [Figure 3c]Figure 14. Loricrin expression staining of 3D epidermis after 9 days of differentiation treated with different calcium concentrations or Ca2+phospholipid complexes. Treatment with CaCl2·2H2O (0.3 mM CaCl2·2H2O on the basal side and 1.1 mM CaCl2·2H2O on the apical side) shows that decreased loricrin production was observed with treatment with 1.1 mM calcium chloride dihydrate solution (c). [Figure 3d] Figure 1 shows loricrin expression staining of 3D epidermis after 9 days of differentiation treated with different calcium concentrations or Ca2+ phospholipid complexes. Treatment with Ca2+ phospholipid complexes (0.3 mM CaCl2·2H2O on the basal side and 0.1% Ca2+ phospholipid complexes on the apical side, preparation similar to sample 6 in Table 1) shows that treatment with Ca2+ phospholipid complexes (d) normalized loricrin expression levels. [Figure 4a] Hematoxylin-eosin staining of human 3D epidermis after 9 days of differentiation treated with different calcium concentrations, phospholipid mixtures, or Ca2+phospholipid complexes, showing control (1.1 mM CaCl2·2H2O on the basal side). [Figure 4b] Hematoxylin-eosin staining of human 3D epidermis after 9 days of differentiation treated with different calcium concentrations, phospholipid mixtures, or Ca2+phospholipid complexes shows reduced calcium concentration (basolateral 0.3 mM CaCl2·2H2O) and impaired epidermis formation was observed at reduced calcium concentration (b). [Figure 4c] FIG. 13 shows hematoxylin-eosin staining of human 3D epidermis after 9 days of differentiation treated with different calcium concentrations, phospholipid mixtures, or Ca2+phospholipid complexes. Treatment with 0.1% phospholipid mixture (apical side, no Ca2+) shows impaired epidermis formation was observed with treatment with 0.1% phospholipid mixture (no Ca2+) on the apical side (c). [Figure 4d]Hematoxylin-eosin staining of human 3D epidermis after 9 days of differentiation treated with different calcium concentrations, phospholipid mixtures, or Ca2+phospholipid complexes shows that treatment with Ca2+phospholipid complexes (basolateral 0.3 mM CaCl2·2H2O and apical 0.1% Ca2+phospholipid complexes, preparation similar to sample 6 in Table 1) supports the formation of a normal epidermis. [Figure 4e] FIG. 10: Hematoxylin-eosin staining of human 3D epidermis after 9 days of differentiation treated with different calcium concentrations, phospholipid mixtures, or Ca2+phospholipid complexes. Quantification of epidermal thickness in a)-d) is shown, with measured epidermal thickness in μm on the ordinate, ***p<0.001 vs. no treatment, **p<0.05 vs. no treatment, ####p<0.0001 vs. reduced Ca2+, ###p<0.001 vs. phospholipid mixture (no Ca2+). [Figure 5a] FIG. 13 shows involucrin staining to determine increased involucrin expression in skin explants treated with placebo (top row, apical side), 0.013% calcium chloride dihydrate solution (bottom row, apical side 0.013% CaCl2·2H2O), or Ca2+ phospholipid complex also containing 0.013% calcium chloride dihydrate (middle row, apical side 0.013% CaCl2·2H2O containing 2% Ca2+ phospholipid complex, preparation similar to sample 6 in Table 1). [Figure 5b] FIG. 1 shows quantification of involucrin expression in a) where the expression of the protein involucrin is shown on the ordinate in % compared to placebo (=100). [Figure 6] FIG. 1 shows the change in various skin parameters compared to the starting values ​​after 7 days of treatment with placebo gel (black bars) or 2% Ca2+phospholipid complex (hatched lines) followed in each case by application of 2% sodium lauryl sulfate for 24 hours (thereby compromising the skin barrier). The change in skin parameters is shown in % on the vertical axis compared to untreated control, *P<0.05 vs. no treatment, **p<0.05 vs. no treatment and placebo, ***p<0.01 vs. no treatment and placebo. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0046] Ca 2+ Formation of phospholipid complex structures: Soy lecithin (containing 30-40% negatively charged phospholipids) was dissolved in alcohol, then transferred to the aqueous phase and subjected to high pressure homogenization, which produced liposomes with a diameter of about 100 nm. To this liposome dispersion, CaCl2·2H2O solutions of different concentrations were added under stirring, and then the mixture was mixed again using the high pressure homogenizer. The obtained products were examined for the formation of stable gel structures and are listed in Table 1 below. For the molar ratio values ​​given in Table 1, it was assumed that the phospholipids had an average molar mass of 762 g / mol in the soy lecithin used. The molar ratios listed were calculated based on the average molar mass of CaCl2·2H2O in the soy lecithin used. 2+ The ratio of Ca to total lecithin (charged and uncharged). 2+ For the ratio of phospholipids to negatively charged lecithin, the molar ratio in the table should be multiplied by 3, assuming that approximately 33% of the phospholipids in lecithin are negatively charged.

[0047] [Table 1]

[0048] In this experiment, stable Ca 2+ It can be seen that the phospholipid complexes (samples 5 and 6) are formed in suitable mixing ratios, where for these lecithins, the molar ratio (total lecithin) is in the range of 0.2:1.0 to 1:1 (i.e., 0.2-1.0), preferably in the range of 0.3:1 to 0.8:1 (i.e., 0.3-0.8), or the molar ratio (negatively charged lecithin) is in the range of 0.6:1 to 3:1 (i.e., 0.6-3), preferably in the range of 1:1 to 2:1 (i.e., 1-2).

[0049] Example 1: 1.2 kg of glycerin, 0.6 kg of ethanol, and 3.56 kg of ultrapure water are mixed at 60°C. 0.3 kg of soy lecithin with a phosphatidylcholine content of 50% is then dispersed therein. After 2 hours, the dispersion is extruded through a high-pressure homogenizer at 1200 bar. This operation is repeated 2-5 times until the average liposome particle size (Z-average) is below 200 nm and the pH is 3-8.

[0050] In the second step, first, 0.3 kg of ultrapure water is charged, 0.0378 kg of CaCl2·2H2O is dissolved in it, and the pH is set to 8.5-9.5.

[0051] The CaCl2 solution is then slowly added to the liposome dispersion while stirring. This results in a CaCl2 solution with increased viscosity. 2+ A lecithin complex is formed (bicone, Fig. 1b). To obtain a homogeneous structure, the entire complex is again extruded through a high-pressure homogenizer at 1200 bar. The final Ca complex with a gelatin-like structure is then extruded through a high-pressure homogenizer at 1200 bar. 2+ The complex can be packaged and presented for use in a topical formulation (aggregated bicone, FIG. 1a).

[0052] Therefore, the molar ratio of divalent calcium to lecithin was finally 0.667, and this example corresponds to sample 6 in Table 1.

[0053] Example 2: 0.3 kg of soybean lecithin with a phosphatidylcholine content of 65% is dissolved at 60° C. with 1.2 kg of ethanol. Following this, this solution is added to 4.16 kg of ultrapure water with stirring. The mixture is then extruded three times through a high-pressure homogenizer at 1200 bar, resulting in a liposome dispersion with an average particle size of 50 nm and a pH of 3-8.

[0054] In the second step, 0.3 kg of ultrapure water is first charged, 0.026 kg of CaCl2·2H2O is dissolved in it, and the pH is set to 8.5.

[0055] The CaCl2 solution is then slowly added to the liposome dispersion while stirring. This results in a CaCl2 solution with increased viscosity. 2+ A phospholipid complex is formed (bicones), which is then homogenized again at 1200 bar to produce a uniform gel structure (aggregated bicones) that can then be used in creams, gels, and other topical products.

[0056] Therefore, the final molar ratio of divalent calcium to lecithin is 0.458.

[0057] Ca 2+ Preparation of further phospholipid conjugates that can be used in combination with the conjugates: The method described can also be used for other divalent ions. Thus, Zn 2+ , Cu 2+ , Mg 2+ , Mn 2+ , Sn 2+ , Mn 2+ , Fe 2+ , or other cations can be incorporated into phospholipid complexes using similar methods, which can increase their biological activity and / or improve their bioavailability.

[0058] Example 3 (Zn 2+ ) 100 g glycerin, 50 g pentylene glycol and 270 g ultrapure water are mixed and heated to 50°C. This is followed by the addition of 25 g sunflower lecithin with a phosphatidylcholine content of 50%. Once the lecithin is well dispersed, the dispersion is homogenized twice at 1200 bar. The resulting liposomes have an average particle size of 150 nm. A second aqueous phase then contains 2.92 g ZnCl2 in 12.73 g water, with a pH of 8.5. This second aqueous phase is then slowly added to the liposome dispersion while stirring. This results in the formation of Zn 2+ Gelatin-like structures with phospholipid complexes are obtained.

[0059] Ca 2+ Application of phospholipid complex (bicone): The Ca prepared in this way 2+ The phospholipid complexes (bicone aggregates), optionally combined with other further complexes, can be applied in concentrated form in the form of gelatin-like structures directly to the skin or can be incorporated into cosmetic or medical formulations. 2+ Since the bicone structure is not stable in water, bicone Ca 2+ The phospholipid complexes are multilayered and macroscopically in the form of aggregated bicones with gelatin-like structures (Figure 1a). These structures can then be formulated into aqueous systems such as oil-in-water emulsions or purely aqueous systems.

[0060] Ca in various cosmetics 2+ Phospholipid Complex Formulation: [Table 2]

[0061] [Table 3]

[0062] [Table 4]

[0063] Ca 2+ Phospholipid Complex Activity: Restoration of epidermal structure in an in vitro skin model, control: CaCl2 Ca 2+We investigated the effect of the phospholipid complexes on the differentiation of 3D reconstructed epidermis. To this end, keratinocyte precursor cells were cultured in low calcium growth medium (TAK-GM, 0.03 mM calcium chloride dihydrate (CaCl2·2H2O) and then used to reconstruct 3D epidermis. For this purpose, the medium was replaced with 3D differentiation medium (TAK-3D) containing 1.1 mM CaCl2·2H2O and the cells were grown in the medium for an additional day. The cultures were then exposed to air to induce differentiation and skin barrier formation and grown for an additional 9 days, changing the medium every 2 days as follows: - Condition 1: 1.1 mM CaCl2·2H2O from the basal side (TAK-3D). This is the standard condition for efficient epidermis formation. - Condition 2: 0.3 mM CaCl2·2H2O (low calcium concentration) from the basal side. Here, the calcium concentration was reduced to result in impaired epidermis formation and therefore to generate an impaired skin barrier. This is the minimum necessary calcium concentration that must be present on the basal side to ensure the formation of an epidermis. - Condition 3: 0.3 mM CaCl2·2H2O basolateral + 1.1 mM CaCl2·2H2O apical. Here, calcium in the form of calcium chloride dihydrate was added additionally to the apical side of the 3D epidermis. The apical application was done for comparability with condition 4. - Condition 4: 0.1% Ca on the apical side to mimic the local application of 0.3 mM CaCl2·2H2O + phospholipid complex on the basolateral side 2+ Phospholipid complex (prepared similarly to sample 6 in Table 1). Basolateral treatment represented systemic treatment with calcium.

[0064] The 3D models were harvested on day 9 and processed for histological analysis. Their stratification was examined under a microscope using hematoxylin-eosin staining and immunohistochemical staining for the epidermal differentiation marker loricrin (Biolegend, Cat. No. 905104). High-resolution photomicrographs (10x magnification) were taken (see Figure 2).

[0065] As expected, standard conditions of 1.1 mM CaCl2·2H2O from the basal side (condition 1) resulted in the formation of an intact 3D epidermis (Fig. 2a). Reducing the calcium concentration to 0.3 mM during the differentiation process (condition 2) strongly impaired the formation of a dense stratified epidermis and led to the formation of vacuoles (Fig. 2b). Treatment of differentiating keratinocytes with 1.1 mM CaCl2·2H2O from the apical side (condition 3, Fig. 2c) further deteriorated the formation of a 3D epidermis. Large vacuoles were observed that no longer ensured the integrity of the epidermis. In contrast, apical treatment with 0.1% Ca 2+ Treatment with the phospholipid complex (condition 4) improved the keratinocyte differentiation process and prevented the formation of vacuoles (Figure 2d).

[0066] Furthermore, portions of the 3D epidermis differentiated under the four conditions mentioned above were examined by immunohistochemical staining for the expression of the differentiation marker loricrin (see Figure 3). Similar to the normal differentiation observed by hematoxylin-eosin staining in Figure 2, high levels of loricrin expression were observed under standard differentiation conditions (condition 1) (Figure 3a, dark staining of the epidermis), and reduced levels of loricrin expression were observed under reduced calcium concentrations (condition 2, Figure 3b). At 0.1% Ca on the apical side, 2+ Treatment with the phospholipid complex (condition 4) increased the expression of the epidermal differentiation marker loricrin compared with treatment with 1.1 mM CaCl2·2H2O on the apical side (condition 3, Fig. 3c) (Fig. 3d).

[0067] This indicates that keratinocyte Ca 2+ Treatment with phospholipid complexes is meant to contribute to efficient differentiation and therefore to the proper formation of a 3D epidermis.

[0068] Restoration of epidermal structure in an in vitro skin model, control: phospholipid mixture In further experiments, we investigated and checked whether the phospholipids present in the calcium phospholipid complexes have an effect on the differentiation of the 3D epidermis. Here, parts of the 3D epidermis were differentiated under the following conditions, grown for 9 days and subsequently stained with hematoxylin-eosin solution: - Condition 1: 1.1 mM CaCl2·2H2O (TAK-3D) on the basal side. As mentioned above, this is the standard condition for efficient epidermis formation. - Condition 2: 0.3 mM CaCl2·2H2O (low calcium concentration) from the basal side. As mentioned above, here the reduced calcium concentration leads to impaired epidermis formation and therefore impaired skin barrier formation. - Condition 3: CaCl2·2H2O on the basolateral side + 0.1% phospholipid mixture (Ca 2+ (No.) Here, "Ca 2+ The phospholipid mixture was prepared as described in the "Formation of phospholipid complex structures" section, but the dispersion was diluted with Ca 2+ without, and preferably with an aqueous solution having a pH in the range of 7.5 to 9 (prepared similarly to Sample 1 in Table 1). Apical administration of the composition, particularly the phospholipid mixture, was performed for comparability with condition 4. - Condition 4: 0.1% Ca on the apical side to mimic the local application of 0.3 mM CaCl2·2H2O + phospholipid complex on the basolateral side 2+ Phospholipid complex (prepared similarly to sample 6 in Table 1).

[0069] The 3D models were harvested on day 9 and processed for histological analysis. The stratification was examined under a microscope using hematoxylin-eosin staining and high-resolution photomicrographs (10x magnification) were taken (see Figure 4). Based on the photomicrographs, the epidermal thickness was analyzed and determined and quantified in micrometers (μm).

[0070] As already shown in previous experiments, standard conditions of 1.1 mM CaCl2·2H2O from the basal side (condition 1) resulted in the formation of an intact 3D epidermis (Fig. 4a). Quantification of the epidermal thickness showed a thickness of 82.5 ± 12.7 μm (Fig. 4e, black bars), which represents the benchmark for an intact stratified epidermis in this experiment.

[0071] Reducing the calcium concentration to 0.3 mM during the differentiation process (condition 2) again strongly impaired the formation of a dense stratified epidermis (Fig. 4b), which was further characterized by a reduction in epidermal thickness to 46.0 ± 2.5 μm (Fig. 4e, open bars).

[0072] Treatment of the differentiating keratinocytes from the apical side with 0.1% phospholipid mixture without calcium (condition 3, Figure 4c) further deteriorated the formation of the 3D epidermis, leading to the formation of vacuoles, as a consequence of which the integrity of the epidermis was no longer ensured, as well as a reduced epidermal thickness of 62.2 ± 9.8 μm (Figure 4e, dotted bars).

[0073] In contrast, apical 0.1% Ca 2+ Treatment with the phospholipid complex (condition 4) improved the keratinocyte differentiation process and prevented the formation of vacuoles (FIG. 4d); furthermore, treatment resulted in increased epidermal thickness (FIG. 4e, hatched bars).

[0074] Ca 2+ Effect of phospholipid complexes on differentiation markers in skin explants Ca 2+ The effect of topical treatment with the phospholipid complex on the expression of involucrin, an epidermal differentiation marker, was investigated in skin explants. Skin explants were treated with either a placebo gel (without calcium), a gel containing 0.013% CaCl2·2H2O, or a gel containing 2% CaCl2·2H2O based on the topical gel formulation of Example 3. 2+ Gel containing phospholipid complexes (prepared similarly to sample 6 in Table 1, corresponding to 0.013% CaCl2·2H2O) was applied topically in Franz diffusion cells with 20 mg of each test article for 24 h. The skin explants were then washed with 3 ml of water and frozen at -80°C. The tissues were fixed in formaldehyde and embedded in paraffin. 5 μm tissue sections were prepared, mounted on glass slides and deparaffinized. The slides were stained with hematoxylin-eosin stain to analyze the tissue structure. The expression of the protein involucrin was determined by immunohistochemical staining and quantified by image analysis. 2% Ca 2+Topical treatment with the phospholipid complexes resulted in 22.0% greater expression of the epidermal differentiation marker involucrin (middle row of Fig. 5a and hatched bars in Fig. 5b) compared to placebo-treated skin explants (top row of Fig. 5a and filled bars in Fig. 5b). In contrast, treatment with the corresponding concentration of CaCl2·2H2O resulted in 91.1% less expression of involucrin (bottom row of Fig. 5a and dotted bars in Fig. 5b) compared to placebo-treated skin explants (top row of Fig. 5a and filled bars in Fig. 5b).

[0075] Ca in the skin under stress 2+ Application of phospholipid complexes In a randomized placebo-controlled clinical trial, Ca 2+ The phospholipid complex was investigated for its skin protection and regeneration efficiency. For this purpose, 20 subjects (age: 23-65 years) with normal skin were enrolled in the study. 2+ A topical gel containing a phospholipid complex (similar to sample 6 in Table 1, containing 0.64% CaCl2·2H2O and 5% phospholipid) was used as the test sample. 2+ The same gel formulation without the phospholipid complex was used as a placebo. Skin parameters measured were cutaneous microcirculation (Periflux PF5000, Perimed, Sweden), transepidermal water loss (TEWL) (Tewameter® TM300, Courage+Khazaka, Germany), and skin color (a * The parameters were redness, Chromameter® CR-400, Minolta, Japan).

[0076] In the skin protection test, these test substances were applied twice a day for 7 days to one forearm of each case. In addition, an area of ​​untreated skin was defined on the forearm to which no product was applied. The above-mentioned skin parameters were then measured, and damage to the skin barrier was obtained by applying 2% sodium lauryl sulfate (SLS) in the form of an occlusive patch to the skin for 24 hours. 2% Ca 2+ To determine the possible protective effect of pretreatment with the phospholipid complex, skin parameters were measured again after removal of the 2% SLS patch and compared between untreated and placebo-treated areas and Ca2+ The phospholipid complex treatment area was compared to the untreated area and the placebo treatment area. 2+ Treatment with the phospholipid complex (Figure 6, hatched bars) resulted in significant reductions in all three parameters. 2+ This means that pretreatment with the phospholipid complex protects the skin from harmful influences and therefore significantly reduces the effects of stress on the skin.

[0077] In addition, Ca 2+ The skin regeneration potential of the phospholipid complex was investigated in the same group of subjects. For this purpose, the abovementioned skin parameters were measured in three untreated skin areas on the forearm, and then a 2% SLS patch was applied in each case for 24 hours. After removal of the SLS patch, measurements of the skin parameters were performed every 2-3 days until the 24th day. Skin redness, skin microcirculation, and regeneration related to TEWL were significantly greater in the 2% Ca complex than in the skin areas treated with placebo gel. 2+ The rate of Ca ion transport was significantly faster in the skin areas treated with the phospholipid complex. 2+ It is demonstrated that the phospholipid complex has a regenerative effect on the skin and accelerates the normalization of the skin barrier after injury.

Claims

1. An active amount of divalent calcium (Ca) for use in the topical treatment of skin with impaired barrier function, including the skin of diabetic or prediabetic patients. ２＋ 1. A pharmaceutical or cosmetic composition comprising: a) a divalent calcium salt of Calcium iodide; b) a divalent calcium salt of Calcium iodide; c) a divalent calcium salt of Calcium iodide; d) a divalent calcium salt of Calcium iodide; and d) a divalent calcium salt of Calcium iodide;

2. The Ca ２＋ The phospholipid complex is ２＋ The composition according to claim 1, characterized in that it is in the form of a complex of with one or two phospholipids.

3. A composition according to claim 1 or 2, characterized in that it is for use in patients with type 2 diabetes.

4. 3. The composition according to claim 1, wherein the phospholipid is a phosphoglyceride in hydrogenated, partially hydrogenated or non-hydrogenated form selected from the group consisting of phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, diphosphatidylglycerol or mixtures of these systems, and / or a lecithin moiety in hydrogenated, partially hydrogenated or non-hydrogenated form.

5. The calcium phospholipid complex, wherein the calcium phospholipid complex is at least partially a calcium complex with a negatively charged phospholipid selected from the group consisting of phosphatidic acid, phosphatidylinositol, or phosphatidylserine, or a combination thereof, and wherein the molar ratio of divalent calcium to phospholipid is: based on the total content of phospholipids in the range of 0.05:1 to 20:1, or in the range of 0.1:1 to 5:1, or in the range of 0.2:1 to 1:1, or in the range of 0.3:1 to 0.8:1, or in the range of 0.1:1 to 30:1, or in the range of 0.3:1 to 10:1, or in the range of 0.4:1 to 5:1, or in the range of 0.5:1 to 1:1, based on the content of negatively charged phospholipids. The composition according to claim 4, characterized in that

6. The composition described in claim 4, characterized in that the calcium phospholipid complex is a calcium complex with negatively charged phosphatidic acid, phosphatidylinositol, and / or phosphatidylserine, and the molar ratio of divalent calcium to negatively charged phospholipid is in the range of 0.2:1 to 1:1 or 0.3:1 to 0.8:

1.

7. A composition described in claim 1 or 2, characterized in that the calcium phospholipid complex is a gel structure in the composition and / or in the calcium phospholipid-containing starting material of the composition.

8. The Ca intended for topical application ２＋ The concentration of the divalent calcium in the phospholipid complex is in the range of 0.001 to 10 weight percent, or in the range of 0.005 to 5.0 weight percent, or in the range of 0.01 to 0.5 weight percent, and CaCl ２ ・2H ２ The weight percent of O is based on the total composition; or the Ca intended for topical application. ２＋ The concentration of the phospholipid in the phospholipid complex is in the range of 0.01 to 30 weight percent, or in the range of 0.05 to 5 weight percent, the weight percent of the phospholipid being based on the total composition.

3. The composition according to claim 1 or 2, characterized in that

9. 3. The composition of claim 1 or 2, wherein the composition is formulated as an ointment, cream, lotion, paste, or tincture for topical application to the skin of the patient.

10. The composition is in the form of a concentrate for the preparation of a topical formulation, wherein the concentration of the phospholipid complex with divalent calcium in the composition is in the range of 0.1 to 100 weight percent, or in the range of 1 to 10 weight percent, and Ca ２＋ 3. The composition of claim 1, wherein the weight percentage of the phospholipid complex is based on the topical composition as a concentrate.

11. A method for the treatment and / or cosmetic treatment of skin with impaired barrier function, including the skin of diabetic or pre-diabetic patients, for the stabilization, prevention, reduction or elimination of dry skin or moisturizing dry skin and / or for restoring barrier function, for the stabilization, prevention, reduction or elimination of skin infections, including those of bacterial origin, due to skin changes or skin irritation caused by impaired barrier function, or for the stabilization, prevention, reduction or elimination of fungal infections, pigmentation disorders (diabetic dermopathy), blistering, itching (diabetic pruritus), redness (including diabetic necrobiosis lipoidica, pseudoacanthosis nigricans, diabetic bullosa, diabetic rubeosis, diabetic scleredema), or wrinkles, or a combination of these indications, 3. A method comprising topically applying the composition of claim 1 or 2 to the skin.

12. 12. The method of claim 11, wherein the composition as a concentrate is formulated as an ointment, cream, lotion, paste, or tincture for topical application to the skin of the patient.

13. A method for preparing the composition of claim 1 or 2, the phospholipid starting material is optionally pre-dissolved or pre-dispersed in an organic solvent, including ethanol, or a mixture of an organic solvent and water, and then provided as a liposome structure in an aqueous medium; The dispersion was then ２＋ Mix with an aqueous solution of A method characterized by:

14. The method of claim 13, wherein the preparation of the liposomes and / or the homogenization is carried out using a high-pressure homogenizer.

15. 14. The method according to claim 13, characterized in that the composition is formulated into an ointment, cream, lotion, paste or tincture using at least one carrier substance and optionally further components.

16. The composition described in claim 1, characterized in that the Ca 2+ phospholipid complex is in the form of a complex of the Ca 2+ and one or two negatively charged phospholipids as a neutral complex structure.

17. The composition described in claim 16, characterized in that the complex is in the form of a neutral conical or biconical structure between two substantially opposing or adjacent, at least partially negatively charged phospholipids, with Ca2+ at the center, or in the form of a neutral aggregate of such structures.

18. The composition of claim 9, further comprising at least one of the following formulation components: thickeners, lubricants, humectants and / or water-retaining agents, surfactants, preservatives, antifoaming agents, waxes, fats, oils, antioxidants and / or substances with antioxidant properties, bactericides, fungicides, fragrances, propellants, dyes, stabilizers, polar and non-polar solvents including water, pigments, UV filters, plant extracts, further active therapeutic or medicinal ingredients, or combinations thereof.

19. The liposome structure has an average particle size of less than 300 nm or less than 200 nm, The method according to claim 13, further comprising mixing the dispersion with an aqueous solution of Ca 2+ , the aqueous solution being set to a pH in the range of 7.5 to 10, or in the range of 8 to 9.

20. The method of claim 13, wherein the step of mixing the dispersion with an aqueous solution of Ca 2+ is followed by homogenization to form a gel.

21. The method of claim 14, wherein the high-pressure homogenizer is operated at a pressure of at least 500 bar, or 1000 bar.

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