COLLAGEN HYDROLYSATE FOR USE AGAINST ATOPIC DERMATITIS

Description

[0001] The present invention relates to collagen hydrolysate for use as an active ingredient in the treatment of neurodermatitis.

[0002] Atopic dermatitis, also known as atopic eczema, is a chronic skin disease that currently affects 1 to 3% of adults and 5 to 20% of children in industrialized countries. The prevalence of atopic dermatitis is generally increasing, with the disease's prevalence estimated to have increased four to six times since the mid-20th century. Although the exact cause of the disease is not yet fully understood, current research suggests it is an autoimmune disease in which both genetic predisposition and environmental influences play a role.

[0003] Atopic dermatitis generally manifests itself in a compromised barrier function of the skin, with the most specific symptoms being reddened, scaly, and sometimes weeping eczema. This is associated with acute or chronic itching, which can be very severe and often persists at night. Itching, in particular, can significantly impair the quality of life of those affected. The severity of atopic dermatitis can be assessed using the SCORAD (Scoring Atopic Dermatitis) index.

[0004] A complete cure for atopic dermatitis is not yet possible, but various approaches are available for treating the symptoms. The focus is on basic skin care in the form of topical treatment with ointments, creams or lotions to stabilize the barrier function and reduce the skin's sensitivity to irritation and the penetration of allergens. If this measure is no longer sufficient in the case of more severe disease or symptoms, corticosteroids are used as anti-inflammatory agents. These preparations with a cortisol effect are often colloquially referred to as cortisone. They are usually applied topically to the affected areas of skin. However, due to the known side effects of corticosteroids, particularly on the skin (such as skin atrophy, pigmentation disorders and ecchymoses), their use should only be short-term.

[0005] DE 10 2012 101911 A1 discloses the use of collagen hydrolysate for the treatment and / or prevention of cellulite.

[0006] RO 115 690 B1 discloses an ointment against acne and seborrhea containing 1 to 3% collagen hydrolysate.

[0007] CN 104 382 943 A discloses the use of a fish scale extract containing collagen peptides for the treatment and / or prevention of psoriasis.

[0008] There is therefore a great need for active ingredients for the treatment of atopic dermatitis and other diseases that are associated with fewer side effects and that can be used as an alternative or in addition to corticosteroids.

[0009] To achieve this objective, the present invention proposes the use of collagen hydrolysate as an active ingredient in the treatment of atopic dermatitis. The corresponding efficacy of collagen hydrolysate is demonstrated in particular by a clinical study in which collagen hydrolysate or a placebo was administered to patients with atopic dermatitis.

[0010] Various beneficial effects of collagen hydrolysate have been known for some time, particularly its use in osteoporosis or joint pain. Positive effects of collagen hydrolysate on skin health have also been described, for example, in international patent application WO 2012 / 065782 A2. Nevertheless, the effectiveness of collagen hydrolysate in the treatment of atopic dermatitis is surprising.

[0011] Collagen hydrolysate, as a degradation product of animal raw materials also used as food, is completely safe for health and has no known harmful side effects. It does not require legal approval as a medicinal product, but can be marketed and used, particularly in the form of a dietary supplement. The present invention encompasses the use of collagen hydrolysate as a dietary supplement, as an over-the-counter (OTC) medicinal product, or as a prescription medicinal product (particularly in combination with other active ingredients).

[0012] The collagen hydrolysate is preferably administered orally. It is known that the peptides in collagen hydrolysate, even at relatively high molecular weights of up to 10,000 Da, are absorbed in the intestine to at least some extent. The preferred amount of orally administered collagen hydrolysate is a daily dose of 2 to 15 g, more preferably 3 to 10 g. A positive effect in atopic dermatitis, for example, has been demonstrated with a daily dose of 5 g of collagen hydrolysate.

[0013] In a preferred embodiment of the invention, the collagen hydrolysate is used as a supportive treatment alongside treatment of the skin disease with a corticosteroid. Even though a corticosteroid often cannot be completely dispensed with to achieve sufficient symptom relief, the administration of collagen hydrolysate at least allows for a reduction in the amount of corticosteroid administered, which already represents a significant improvement for those affected with regard to its side effects.

[0014] The corticosteroid, alongside which the collagen hydrolysate can be used according to the invention, is generally administered topically, particularly in the form of an ointment. Alternatively, oral administration of corticosteroids is also possible. In this latter case, according to a further embodiment of the invention, a composition containing the collagen hydrolysate and the corticosteroid can be administered, i.e., in the form of a combination preparation.

[0015] The corticosteroid is preferably a glucocorticoid, such as hydrocortisone. A variety of glucocorticoids are known, which are divided into four classes, from "weakly effective" (Class I) to "very potent" (Class IV).

[0016] A further aspect of the disclosure concerns collagen hydrolysate for use as an active ingredient to mitigate the side effects of corticosteroids, particularly their side effects on the skin, such as skin atrophy, pigmentation disorders, and ecchymoses. Corresponding evidence also emerges from the aforementioned clinical study, which is discussed in detail below.

[0017] The above-mentioned side effects of corticosteroids (especially glucocorticoids) are at least partly caused by a reduction in collagen biosynthesis by skin cells (fibroblasts) (see, for example, Oikarinen et al., Journal of Investigative Dermatology 98 (1992) 220-225). Cell experiments in vitro have surprisingly shown (see below) that this reduction in collagen biosynthesis can be almost completely compensated by the presence of collagen hydrolysate.

[0018] The collagen hydrolysate for use according to the invention typically has an average molecular weight of 500 to 15,000 Da, preferably of 1,000 to 8,000 Da, more preferably of 1,500 to 5,000 Da, most preferably of 1,800 to 2,200 Da. These figures always refer to the weight-average molecular weight, which can be determined in particular by gel permeation chromatography.

[0019] The collagen hydrolysate is preferably produced by enzymatic hydrolysis of a collagen-containing starting material. Endopeptidases or exopeptidases of microbial or plant origin are particularly used for this hydrolysis. By appropriately selecting the peptidases and hydrolysis conditions, collagen hydrolysates can be produced in the desired molecular weight range.

[0020] The collagen-containing starting material is generally selected from the skin or bones of vertebrates, preferably mammals or birds, and in particular from the skin of cattle or pigs (beef split skin or pork rind, respectively). Alternatively, the collagen-containing starting material can be selected from the skin, bones, and / or scales of fish, particularly cold- or warm-water fish.

[0021] The collagen hydrolysate can be produced either in a one-step process from these starting materials or via the intermediate step gelatin, in which case both type A and type B gelatin can be used.

[0022] Preferably, the collagen hydrolysate is produced by the sequential action of at least two endoproteases with different specificities, in particular of at least two different metalloproteases and / or serine proteases, i.e. proteases that cleave the amino acid sequence of the collagen molecules before or after specific amino acids. Advantageously, the metalloproteases and / or serine proteases are enzymes from the microorganisms. Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Aspergillus oryzae and Aspergillus melleus.

[0023] By selecting suitable endoproteases, not only can a specific molecular weight distribution of the collagen hydrolysate be achieved, but the nature of the amino acids at the termini of the peptides contained in the hydrolysate can also be influenced. In this regard, for example, it is preferred that at least 50% of the N-terminal amino acids of the collagen hydrolysate be hydrophobic amino acids, especially alanine, leucine, and isoleucine.

[0024] As an alternative to enzymatic hydrolysis, the collagen hydrolysate can be produced by recombinant gene expression within the scope of the invention. By using natural collagen sequences, particularly from cattle or pigs, and expressing them in genetically modified cells (e.g., yeast, bacteria, or plant cells, especially tobacco), products can be produced that are essentially identical to the hydrolysis products of the corresponding collagen-containing raw materials. This makes it possible to obtain a narrower or precisely specified molecular weight distribution.

[0025] The effectiveness of collagen hydrolysate in the treatment of atopic dermatitis is further explained in the clinical study described below.

[0026] Furthermore, cell experiments in vitro described which demonstrate the effectiveness of collagen hydrolysate in mitigating the side effects of corticosteroids.

[0027] The figures show: Figure 1: A bar chart showing corticosteroid and skin care ointment consumption during the study period; Figure 2: A bar chart showing the development of skin pH during the study period; Figure 3: A bar chart showing the change in the "interference with housework / errands due to itching" index during the study period; Figure 4: Bar charts showing the biosynthesis of type I collagen and proteoglycans by fibroblasts; Figure 5: Bar charts showing the biosynthesis of type I collagen and proteoglycans by chondrocytes; and Figure 6: Bar charts showing the biosynthesis of type I collagen and proteoglycans by osteoblasts. 1. Clinical study 1.1 Study design

[0028] The clinical study was conducted with thirty subjects (15 men and 15 women) diagnosed with atopic dermatitis, with the severity of atopic dermatitis according to the SCORAD index ranging from 15 to 40 for all subjects. This was a randomized, double-blind, placebo-controlled study, meaning that the subjects were randomly assigned to a treatment group and a placebo group of 15 people each. Neither the subjects nor the treating staff were informed of the assignment of the individual subjects.

[0029] The subjects in the treatment group received a daily dose of 5 g of collagen hydrolysate throughout the entire 12-week study period. This product was Verisol B, marketed by the applicant. It is a collagen hydrolysate produced by enzymatic hydrolysis of bovine collagen with an average molecular weight of approximately 2,000 Da. The manufacturing process for Verisol B is essentially the same as that described in WO 2012 / 065782 A2.

[0030] The subjects in the placebo group received a daily dose of 5 g of maltodextrin instead of the collagen hydrolysate, with the collagen hydrolysate and the placebo being indistinguishable in terms of packaging, texture and taste.

[0031] During the study, participants were free to decide whether and to what extent they used a skin care ointment (Ungentum leniens) or a corticosteroid-containing ointment (triamcinolone acetonide) to relieve their symptoms. The amounts of skin care ointment and corticosteroid used were recorded for each individual participant during the study period. 1.2 Consumption of skin care ointment and corticosteroid

[0032] A notable finding of the study is that the consumption of both skin care ointment and corticosteroid-containing ointment was significantly lower in the study group than in the placebo group. This applied to both the first third (4 weeks) of the study period and the period from weeks 5 to 12, and was particularly pronounced for corticosteroid consumption.

[0033] In the Figure 1The corresponding results are presented as a bar chart, ie the average consumption of corticosteroid-containing ointment and skin care ointment in grams in the two groups for the respective periods.

[0034] This result demonstrates the effectiveness of collagen hydrolysate in the treatment of atopic dermatitis. Since the subjects were free to choose the amount of corticosteroid and skin care ointment as needed, the lower amount required in the study group can only be explained by the fact that some of the symptoms could be directly alleviated by the orally administered collagen hydrolysate. 1.3 Measuring the pH of the skin

[0035] The pH of the skin, an indicator of its barrier function, was measured in all subjects at the start of the study, after four weeks, and after twelve weeks using a skin pH meter (Courage + Khazaka electronic GmbH). Measurements were taken on the upper arm, both on healthy and lesion-affected skin areas.

[0036] The results are in the Figure 2 shown as a bar chart.

[0037] In the treatment group, the pH of lesional skin tended to approach the pH of healthy skin, ranging from 5.3 to 5.6, during the study period. In contrast, in the placebo group, the pH of both healthy skin and lesional skin was abnormally elevated after 12 weeks.

[0038] This result suggests that oral administration of collagen hydrolysate at least partially counteracts the side effects of corticosteroids on the skin. 1.4 Transepidermal water loss and skin moisture

[0039] During the study period, measurements of transepidermal water loss (TEWL) were also performed using a Tewameter and skin hydration using a Corneometer (both from Courage + Khazaka electronic GmbH). Both parameters, like pH, are indicators of the skin's barrier function.

[0040] As a result, no significant differences were found in these two parameters between the treatment group and the placebo group, either on healthy or lesional skin sites. This indirectly suggests that despite the higher dose of corticosteroid used in the placebo group and its associated side effects, no deterioration in these parameters occurred, which could be attributed to the effect of the collagen hydrolysate. 1.5 Assessment of itching

[0041] During the study period, participants completed a 5D itching assessment using a scheme known as "5-D Pruritus." This scheme assesses the itch itself quantitatively and qualitatively (duration, severity, and tendency), its location on the body (distribution), and the impact of the itch on the following areas of life: sleep; leisure / social activities; housework / errands; and work / school.

[0042] Regarding the assessment criterion "Impairment of housework / chores," the treatment group showed a significant improvement in the index during the study period compared to the placebo group. The corresponding results are shown in the bar chart of the Figure 3 (mean index values ​​at baseline, after four weeks, and after twelve weeks). This result directly supports the effectiveness of collagen hydrolysate in relieving itching.

[0043] Regarding the remaining evaluation criteria, no significant differences in the respective indices were found between the study group and the placebo group. However, considering that significantly more corticosteroid was consumed in the placebo group, this finding also indirectly allows the conclusion that the effect of the corticosteroid could be at least partially replaced by collagen hydrolysate. 2. Cell experiments in vitro 2.1 Cell experiments with fibroblasts

[0044] Topical treatment with corticosteroids, especially over longer treatment periods, is associated with negative side effects such as skin atrophy. A major cause of these side effects is the reduction in the biosynthesis of collagen, as well as other important matrix proteins such as proteoglycans, by fibroblasts.

[0045] Cell experiments with human fibroblasts were conducted to investigate whether collagen hydrolysate in combination with glucocorticoids influences matrix protein biosynthesis. These experiments used the same collagen hydrolysate (Verisol B) as in the clinical study described above.

[0046] Primary human fibroblasts were cultured in HAM's F12 medium supplemented with 10% fetal calf serum, 20 U / ml penicillin-streptomycin, 50 µg / ml patricin, 0.05 mg / ml ascorbic acid, and 0.15 mg / ml glutamine. After reaching 80% cell confluence, the culture medium was replaced in three different batches with fresh medium supplemented with 0.5 mg / ml collagen hydrolysate, 0.05 mg / ml of a glucocorticoid (dexamethasone), or a combination of both. A control batch was further cultured in the culture medium without any additives.

[0047] After a further seven days of cultivation, the amount of extracellular matrix proteins synthesized by the fibroblasts was determined using the Sircol Soluble Collagen Assay and the determination of proteoglycans using the Glycosaminoglycan Assay Blyscan (Biocolor Ltd, UK), each according to the manufacturer's instructions.

[0048] The results are shown in the bar charts of the Figure 4 shown, where Figure 4A the biosynthesis of collagen type I and Figure 4Bthe biosynthesis of proteoglycans. These are the mean values ​​from eight experiments, expressed in relation to the control. In both cases, it is initially evident that the biosynthesis of collagen and proteoglycans is, as expected, greatly reduced by glucocorticoids. However, this reduction can be completely compensated by additional supplementation with collagen hydrolysate. Collagen hydrolysate alone has a slightly stimulating effect on biosynthesis.

[0049] These results provide further evidence that the side effects of corticosteroids, such as skin atrophy, which are caused by a reduction in the biosynthesis of matrix proteins in skin cells, can be reduced by the administration of collagen hydrolysate. 2.2 Cell experiments with chondrocytes

[0050] The same cell experiments were also performed with human chondrocytes (cartilage cells). Cell cultivation and determination of type I collagen and biproteoglycans were performed as in the fibroblast experiments described above, with the difference that a collagen hydrolysate from bovine bone gelatin with an average molecular weight of approximately 4,000 Da was used.

[0051] The results are shown in the bar charts of the Figure 5 shown, where Figure 5A the biosynthesis of collagen type I and Figure 5B the biosynthesis of proteoglycans. These are the average values ​​from three experiments. Here, too, it can be seen that the reduction in biosynthesis caused by the glucocorticoid can be essentially completely compensated by the addition of collagen hydrolysate. 2.3 Cell experiments with osteoblasts

[0052] Similar experiments were also conducted with human osteoblasts (bone cells), and the assay procedure and determination of collagen type I and proteoglycans was carried out as in the experiments with chondrocytes (including the collagen hydrolysate with 4,000 Da). The bar charts of the Figure 6 show that also in the case of osteoblasts the reduction in the biosynthesis of collagen type I ( Figure 6A ) and proteoglycans ( Figure 6B ) by the glucocorticoid can be essentially completely compensated by the addition of collagen hydrolysate.

[0053] The results with chondrocytes and osteoblasts show that the beneficial effects of collagen hydrolysate in combination with corticosteroids are not limited to the skin, but that side effects in other tissue types can also be expected to be alleviated.

Claims

1. A collagen hydrolysate for use as an active substance in the treatment of neurodermatitis.

2. A collagen hydrolysate for use according to claim 1, wherein the collagen hydrolysate is administered orally, preferably in a daily dose of from 2 to 15 g, more preferably from 3 to 10 g.

3. A collagen hydrolysate for use according to claim 1 or 2, wherein the collagen hydrolysate is used for supportive treatment in addition to a treatment of the neurodermatitis.

4. A collagen hydrolysate for use according to claim 3, wherein the administration of collagen hydrolysate allows a reduction of the administered amount of corticosteroid.

5. A collagen hydrolysate for use according to one of claims 3 to 4, wherein the corticosteroid is administered topically, in particular in the form of an ointment, or wherein the corticosteroid is administered orally, in particular in the form of a composition which contains the collagen hydrolysate and the corticosteroid.

6. A collagen hydrolysate for use according to one of claims 3 to 5, wherein the corticosteroid is a glucocorticoid, for example hydrocortisone.

7. A collagen hydrolysate for use according to one of the preceding claims, wherein the collagen hydrolysate has a mean molecular weight of from 500 to 15,000 Da, preferably from 1,000 to 8,000 Da, more preferably from 1,500 to 5,000 Da, most preferably from 1,800 to 2,200 Da.

8. A collagen hydrolysate for use according to one of the preceding claims, wherein the collagen hydrolysate is produced by enzymatic hydrolysis of a collagen-containing starting material.

9. A collagen hydrolysate for use according to claim 8, wherein the collagen-containing starting material is selected from skin or bone of vertebrates, preferably of mammals, birds or fish, in particular from skin of cattle or pigs.

10. A collagen hydrolysate for use according to claim 8 or 9, wherein the collagen hydrolysate is produced by the successive action of at least two endoproteases having a different specificity, in particular of at least two different metalloproteases and / or serine proteases.

11. A collagen hydrolysate for use according to claim 10, wherein the metalloproteases and / or serine proteases are selected from enzymes from the microorganisms Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Aspergillus oryzae and Aspergillus melleus.

12. A collagen hydrolysate for use according to one of the preceding claims, wherein at least 50% of the N-terminal amino acids of the collagen hydrolysate are hydrophobic amino acids, in particular alanine, leucine and isoleucine.

13. A collagen hydrolysate for use according to one of claims 1 to 7, wherein the collagen hydrolysate is produced by recombinant gene expression.