Collagen hydrolysate for use in skin disorders and intestinal disorders

Abstract

The present invention relates to a collagen hydrolysate for use as an active substance in the treatment of inflammatory skin and bowel disorders and / or skin disorders and bowel disorders associated with inflammation.

Description

Technical Field

[0001] The present invention relates to a collagen hydrolysate for use as an active substance in the treatment of inflammatory skin disorders and intestinal disorders and / or skin disorders, and intestinal disorders accompanied by inflammation.

[0002] The present invention also relates to a collagen hydrolysate for use as an active substance for reducing the side effects of corticosteroids, especially their side effects on the skin.

Background Art

[0003] Neurodermatitis, also called atopic eczema, is a chronic skin disorder that currently affects 1-3% of adults and 5-20% of children in developed countries. The incidence of neurodermatitis is generally increasing, and since the mid-20th century, the incidence of the disorder has increased 4 to 6 times. The exact cause of the disorder is not yet fully understood, but according to the current state of research, it is an autoimmune disorder in which both genetic predisposition and environmental influences play a role.

[0004] Neurodermatitis generally appears in the form of a disorder of the skin barrier function, and its specific symptoms are particularly redness, scaly, and sometimes weeping eczema. They are associated with acute or chronic itching, which can be very severe and sometimes continue throughout the night. In particular, itching can significantly impair the quality of life of the affected individual. The severity of neurodermatitis can be evaluated according to the SCORAD (Scoring Atopic Dermatitis) index.

[0005] So far, atopic dermatitis has not been completely curable, but various approaches can be used to treat the symptoms. The main focus here 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 allergen infiltration. In cases of more severe symptoms of the disorder or condition, especially when corticosteroids are used as a treatment means as anti-inflammatory agents, those preparations having a cortisone effect are commonly referred to as cortisone in everyday language. They are usually applied topically to the skin area of the affected part, but are only suitable for short-term application due to the known side effects of corticosteroids, especially side effects on the skin (e.g., skin atrophy, pigmentation disorders and purpura).

Summary of the Invention

Problems to be Solved by the Invention

[0006] Therefore, there is a great need for active substances for the treatment of atopic dermatitis and other disorders that have few side effects and can be used instead of or in addition to corticosteroids.

Means for Solving the Problems

[0007] To solve this problem, the present invention provides the use of collagen hydrolysate as an active substance for the treatment of inflammatory skin disorders and intestinal disorders and / or skin disorders, and intestinal disorders accompanied by inflammation. The appropriate effectiveness of collagen hydrolysate has been revealed in particular from clinical studies in which collagen hydrolysate or placebo was administered to patients with atopic dermatitis.

[0008] Various beneficial effects of collagen hydrolysate, especially the use of collagen hydrolysate in osteoporosis or joint complaints, have been known for a relatively long time. The positive effect of collagen hydrolysate on skin health has also already been described, for example, in international patent application WO2012 / 065782A2. Nevertheless, the effectiveness of collagen hydrolysate in the treatment of atopic dermatitis and other inflammatory disorders is surprising.

[0009] Collagen hydrolysis as a by - product of animal - derived substances used also as food is a product that is completely harmless from a health perspective and has no known harmful side effects. It does not require legal approval as a drug and, in particular, can be sold and used in the form of dietary supplements. The use of collagen hydrolysates as dietary supplements, over - the - counter (OTC) drugs, or prescription drugs (especially in combination with other active substances) is included within the scope of the present invention.

[0010] Collagen hydrolysates are preferably administered orally. The peptides of collagen hydrolysates are known to be re - absorbed by the intestine to at least some extent, even with a relatively high molecular weight of up to 10,000 Da. For a daily dose, the preferred amount of orally administered collagen hydrolysates is 2 - 15 g, more preferably 3 - 10 g. The intestinal effect in the case of atopic dermatitis could be demonstrated with a daily dose of 5 g of collagen hydrolysates. BRIEF DESCRIPTION OF THE DRAWINGS

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Mode for Carrying Out the Invention

[0017] According to a preferred embodiment of the present invention, the collagen hydrolyzate is used as a supportive therapy in addition to the treatment of skin or intestinal disorders with corticosteroids. Although it is often impossible to completely omit corticosteroids and furthermore to sufficiently relieve the symptoms, the administration of the collagen hydrolyzate makes it possible to at least reduce the dosage of corticosteroids, which in itself already brings about a significant improvement in the side effects of the affected individuals.

[0018] The corticosteroids that can be used together with the collagen hydrolyzate according to the present invention are generally administered topically, especially in the form of ointments. On the other hand, oral administration of corticosteroids is also possible. In this latter case, according to a further embodiment of the present invention, the composition containing the collagen hydrolyzate and the corticosteroid can be administered, i.e., in the form of a combined preparation.

[0019] The corticosteroid is preferably a glucocorticoid, such as hydrocortisone. A number of glucocorticoids classified from "almost ineffective" (class I) to "very effective" (class IV) are known.

[0020] A preferred embodiment of the present invention relates to a collagen hydrolyzate for use as an active substance in the treatment of atopic dermatitis. Further inflammatory skin disorders and / or skin disorders accompanied by inflammation for which treatment or supportive therapy with collagen hydrolyzate is possible are psoriasis, rosacea, chronic pruritus, acne, cellulitis and skin porosity.

[0021] The present invention also includes collagen hydrolysates for use in the treatment of inflammatory bowel diseases and / or bowel diseases associated with inflammation. There is a similarity between inflammatory skin disorders and inflammatory bowel disorders as long as the intestinal wall contains collagen fibers as structural proteins and represents a barrier between the actual body and the "outer region". Bowel diseases enabling the use of collagen hydrolysates within the scope of the present invention are, in particular, Crohn's disease and ulcerative colitis. They are chronic inflammatory bowel diseases that can be symptomatically treated with glucocorticoids, respectively.

[0022] A further aspect of the present invention relates to collagen hydrolysates for use as active substances for reducing the side effects of corticosteroids, in particular their side effects on the skin, such as skin atrophy, pigmentation disorders and purpura. Corresponding information can also be found in the clinical studies already mentioned, which will be explained in more detail below.

[0023] The above side effects of corticosteroids (especially glucocorticoids) are at least partly caused by a decrease in collagen biosynthesis by skin cells (fibroblasts) (see, for example, Oikarinen et al., Journal of Investigative Dermatology 98 (1992) 220-225). Cell tests in vitro have surprisingly found that the decrease in collagen biosynthesis can be almost completely compensated for by the presence of collagen hydrolysates (see below).

[0024] Collagen hydrolysates for use according to the present invention typically have an average molecular weight of 500 to 15,000 Da, preferably 1,000 to 8,000 Da, more preferably 1,500 to 5,000 Da, and most preferably 1,800 to 2,200 Da. In those statements, the weight average molecular weight, which can be determined in particular by gel permeation chromatography, is always meant.

[0025] Collagen hydrolyzates are preferably produced by enzymatic hydrolysis of collagen-containing starting materials. In particular, endopeptidases or exopeptidases of microbial or plant origin are used for this hydrolysis. Collagen hydrolyzates in a desired molecular weight range can be produced by appropriate selection of peptidases and hydrolysis conditions.

[0026] Collagen-containing starting materials are generally selected from the skin or bone of vertebrates, preferably mammals or birds, and in particular the skin of cows or pigs (cowhide or pigskin, respectively). On the other hand, collagen-containing starting materials can be selected from the skin, bone and / or scales of fish, especially cold-water fish or tropical fish.

[0027] Collagen hydrolyzates can be produced from these starting materials in a one-step process or via intermediate gelatin; in the latter case, both type A and type B gelatin can be used.

[0028] Collagen hydrolyzates are preferably produced by the sequential action of at least two endoproteases having different specificities, in particular at least two different metalloproteases and / or serine proteases, i.e., proteases that cleave the amino acid sequence of the collagen molecule before and after specific amino acids. The metalloproteases and / or serine proteases are advantageously enzymes derived from the microorganisms Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Aspergillus oryzae and Aspergillus melleus.

[0029] By appropriate selection of endoproteases, not only can collagen hydrolyzates with a specific molecular weight distribution be obtained, but also the type of amino acids at the ends of the peptides contained in the hydrolyzates is affected. In this regard, for example, it is preferred that at least 50% of the N-terminal amino acids of the collagen hydrolyzate are hydrophobic amino acids, especially alanine, leucine and isoleucine.

[0030] Instead of enzymatic hydrolysis, collagen hydrolysates can be produced by recombinant gene expression within the scope of the present invention. In particular, by using natural collagen sequences from cows or pigs and their expression in genetically modified cells (e.g., yeast, bacteria or plant cells, especially tobacco), products substantially identical to the hydrolysis products of the corresponding collagen-containing materials can be produced. Here, it is possible to obtain a narrower or more precisely defined molecular weight distribution.

[0031] The effectiveness of collagen hydrolysates in the treatment of atopic dermatitis will be explained in more detail based on the clinical studies described below.

[0032] Furthermore, in vitro cell tests demonstrating the effectiveness of collagen hydrolysates in reducing the side effects of corticosteroids are described.

Example

[0033] 1. Clinical trial 1.1 Research design A clinical trial was conducted with 30 subjects (15 men and 15 women) diagnosed with atopic dermatitis. According to the SCORAD index, the severity of atopic dermatitis was in the range of 15 - 40 for all subjects. This study was a randomized, double-blind, placebo-controlled trial, i.e., the subjects were divided into a treatment group and a placebo group, each containing 15 individuals, and neither the subjects nor the treatment staff were aware of the group assignment of each individual subject.

[0034] Subjects in the treatment group were administered 5 g of collagen hydrolysate daily for the entire 12-week study period. This was Verisol B, a product distributed by the applicant, which was a collagen hydrolysate produced by enzymatic hydrolysis of bovine collagen with an average molecular weight of approximately 2,000 Da. The manufacturing method for Verisol B substantially corresponded to the manufacturing method described in WO2012 / 065782A2.

[0035] In the placebo group, the subjects were administered 5 g of maltodextrin daily instead of the collagen hydrolyzate, and there was no difference in terms of packaging, texture or taste between the collagen hydrolyzate and the placebo.

[0036] During the study, the subjects could freely choose whether to apply a skin care ointment (unguentum leniens) or a corticosteroid-containing ointment (triamcinolone acetonide) and the amount to apply in order to relieve symptoms. The use of skin care ointments and corticosteroids for each individual subject was measured during the study period.

[0037] 1.2 Use of skin care ointment and corticosteroid The notable result of the study is that the use of both skin care ointments and corticosteroid-containing ointments in the investigation group was significantly less than in the placebo group. This applies to both the first third (4 weeks) of the study period and the period from week 5 to week 12, and in both cases it is particularly pronounced with the use of corticosteroids.

[0038] Figure 1 shows the corresponding results in the form of a bar graph, namely the average usage amounts in grams for each period in both groups of corticosteroid-containing ointments and skin care ointments.

[0039] This result indicates the effectiveness of collagen hydrolyzate in atopic dermatitis. Since the amounts of corticosteroids and skin care ointments can be freely selected by the subjects as needed, the fact that the required amounts in the investigation group were less can only be explained by the fact that some symptoms could be directly alleviated by the orally administered collagen hydrolyzate.

[0040] 1.3 Measurement of skin pH value The pH value of the skin, which is an indicator of the barrier function, was measured in all test subjects at the start of the study, after 4 weeks, and after 12 weeks using a skin pH meter (Courage + Khazaka electronic GmbH). The measurements were performed in the area of the upper arm, more specifically, in both the healthy skin area and the skin area affected by the lesion.

[0041] The results are shown in Figure 2 in the form of a bar graph.

[0042] In the treatment group, the pH value of the skin area with lesions tended to approach the pH value of healthy skin in the range of 5.3 - 5.6 during the study period. In contrast, in the placebo group, the pH values of both the healthy skin and the skin area with lesions increased abnormally after 12 weeks.

[0043] This result leads to the conclusion that oral administration of collagen hydrolysate at least partially cancels out the side effects of corticosteroids on the skin.

[0044] 1.4 Trans-epidermal water loss and skin moisture Measurements of transepidermal water loss (TEWL) using a Tewameter and skin moisture using a Corneometer (both from Courage + Khazaka electronic GmbH) were performed during the study period. Both parameters are indicators of the skin barrier function, similar to the pH value.

[0045] No significant differences between the treatment group and the placebo group were found in the results of these two parameters, either in the healthy skin area or in the skin area affected by the lesion. This indirectly leads to the conclusion that, despite the larger amount of corticosteroids used in the placebo group and their effects, no deterioration of the parameters that could be attributed to the effect of collagen hydrolysate occurred.

[0046] 1.5 Evaluation of itching The evaluation of itching was conducted on the subjects during the study period according to a model called the "5D Pruritus Scale". According to this model, itching is evaluated both quantitatively and qualitatively (in terms of duration, severity, and tendency) with respect to its localization (distribution) on the body and also with respect to the impairment of the following areas of life as a result of itching: sleep; free time / social activities; household chores / errands; work / school.

[0047] Regarding the evaluation criterion of "impairment of household chores / errands", a significant improvement in the indicators was observed in the treatment group during the study period, in contrast to the placebo group. The corresponding results are shown in the bar graph of Figure 3 (average values of the indicators at the start of the study, after 4 weeks, and after 12 weeks). This result is a direct reflection of the effectiveness of collagen hydrolysate in relieving itching.

[0048] Regarding the other evaluation criteria, there were no significant differences in various indicators between the investigation group and the placebo group. However, considering the fact that more corticosteroids were used in the placebo group, this finding also indirectly leads to the result that the effect of corticosteroids may be at least partially replaced by collagen hydrolysate.

[0049] 2. In vitro cell test 2.1 Cell test by fibroblasts Topical treatment with corticosteroids is associated with longer treatment periods, especially with true side effects such as skin atrophy. The main cause of these side effects is the reduction in the biosynthesis of collagen by fibroblasts and / or the biosynthesis of other important matrix proteins such as proteoglycans.

[0050] Cell tests were carried out using human fibroblasts to investigate whether collagen hydrolysate in combination with glucocorticoids affects the biosynthesis of matrix proteins. The same collagen hydrolysate (Verisol B) as in the clinical trials was used in those tests.

[0051] Primary human fibroblasts were cultured in HAM's F12 medium supplemented with 10% fetal bovine 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 medium was replaced with fresh medium supplemented with 0.5 mg / ml collagen hydrolysate, 0.05 mg / ml glucocorticoid (dexamethasone), or a combination of both components in three different batches. The control batch was further cultured in the medium without addition.

[0052] After a further 7-day culture, the amount of extracellular matrix proteins synthesized by the fibroblasts was measured. Type I collagen was measured by the "Sircol soluble collagen assay", and proteoglycan was measured by the "Glycosaminoglycan assay Blyscan" (Biocolor Ltd, Great Britain), in both cases following the manufacturer's instructions.

[0053] The results are shown as bar graphs in Figure 4, where Figure 4A relates to the biosynthesis of type I collagen and Figure 4B relates to the biosynthesis of proteoglycan. In both cases, the average value of 8 tests is specified in relation to the control batch. In either case, it was first found that the biosynthesis of collagen and proteoglycan was significantly reduced by the glucocorticoid as expected. However, this reduction can be completely compensated by the additional supplementation of collagen hydrolysate. Collagen hydrolysate has a mild effect on biosynthesis by itself.

[0054] These results further show that the side effects of corticosteroids, such as skin atrophy, caused by the reduction in the biosynthesis of matrix proteins in skin cells, can be reduced by the administration of collagen hydrolysate.

[0055] 2.2 Cell test by chondrocytes The same cell tests were also performed on human chondrocytes (cartilage cells). Cell culture and measurement of type I collagen and proteoglycan were carried out in the same manner as in the above tests using fibroblasts, but a collagen hydrolyzate from bovine bone gelatin having an average molecular weight of about 4,000 Da was used.

[0056] The results are shown in the bar graphs of FIG. 5, where FIG. 5A shows the biosynthesis of type I collagen and FIG. 5B shows the biosynthesis of proteoglycan. In both cases, the average values from three tests are shown. Here too, it has been found that the decrease in biosynthesis caused by glucocorticoids can be substantially compensated for by the addition of collagen hydrolyzate.

[0057] 2.3 Cell test by osteoblasts Corresponding tests were also performed on human osteoblasts (bone cells); the tests were carried out in the same manner as the tests using chondrocytes (including 4,000 Da collagen hydrolyzate), and type I collagen and proteoglycan were measured. The bar graphs of FIG. 6 show that also in the case of osteoblasts, the decrease in biosynthesis of type I collagen (FIG. 6A) and proteoglycan (FIG. 6B) by glucocorticoids can be substantially completely compensated for by the addition of collagen hydrolyzate.

[0058] The results for chondrocytes and osteoblasts indicate that the beneficial effect of collagen hydrolyzate in combination with corticosteroids is not limited to the skin and that a reduction in side effects can also be expected for other tissue types.

Claims

1. A pharmaceutical composition for the treatment of neurodermatitis, containing hydrolyzed collagen as an active substance, wherein the pharmaceutical composition is used for supportive therapy in addition to the treatment of neurodermatitis with corticosteroids, and the hydrolyzed collagen has an average molecular weight of 1,500 to 5,000 Da.

2. The pharmaceutical composition according to claim 1, wherein the hydrolyzed collagen is orally administered in a daily dose of 2 to 15 g.

3. The pharmaceutical composition according to claim 2, wherein the hydrolyzed collagen is orally administered in a daily dose of 3 to 10 g.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the administration of the hydrolyzed collagen enables a reduction in the corticosteroid dosage.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the corticosteroid is topically applied.

6. The pharmaceutical composition according to any one of claims 1 to 4, wherein the corticosteroid is orally administered in the form of a composition containing hydrolyzed collagen and corticosteroid.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the corticosteroid is a glucocorticoid.

8. The pharmaceutical composition according to any one of claims 1 to 7, wherein the hydrolyzed collagen has an average molecular weight of 1,800 to 2,200 Da.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the hydrolyzed collagen is produced by enzymatic hydrolysis of a collagen-containing starting material.

10. The pharmaceutical composition according to claim 9, wherein the collagen-containing starting material is selected from the skin of vertebrates.

11. The pharmaceutical composition according to claim 9 or 10, wherein the hydrolyzed collagen is produced by the sequential action of at least two metalloproteases and / or serine proteases having different specificities.

12. The pharmaceutical composition according to claim 11, wherein the metalloprotease and / or serine protease is selected from the enzymes produced by the microorganisms Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Aspergillus oryzae and Aspergillus melleus.

13. The pharmaceutical composition according to any one of claims 1 to 12, wherein at least 50% of the N-terminal amino acids of the collagen hydrolyzate are hydrophobic amino acids.

14. The pharmaceutical composition according to any one of claims 1 to 8, wherein the collagen hydrolyzate is prepared from collagen produced by recombinant gene expression.

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