HYDROLYZED COLLAGEN FOR USE WITH SKIN AND INTESTINAL DISORDERS.

Abstract

The present invention relates to a collagen hydrolysate for use as an active substance in the treatment of neurodermatitis.

Description

Hydrolyzed collagen for use with skin and intestinal disorders DESCRIPTIVE MEMORANDUM The present invention relates to 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. The invention also relates to collagen hydrolysate for use as an active substance to mitigate the side effects of corticosteroids, specifically their side effects on the skin. Neurodermatitis, also known as atopic eczema, is a chronic skin disorder that currently affects 1 to 3% of the adult population and 5 to 20% of the child population in industrialized countries. Overall, the incidence of neurodermatitis is increasing, and since the mid-20th century, there has been a four- to six-fold increase in the occurrence of the disorder. Although the exact cause of the disorder is not yet fully understood, according to current research, it is an autoimmune disorder in which both genetic predisposition and environmental influences play a role. In general, neurodermatitis manifests as impaired skin barrier function. Specific symptoms include redness, scaling, and sometimes oozing eczema. These are associated with acute or chronic itching, which can be very intense and sometimes persists throughout the night. This itching can significantly impair the quality of life of affected individuals. The severity of neurodermatitis can be assessed using the SCORAD (Score for Atopic Dermatitis) index. To date, there is no complete cure for neurodermatitis; however, several approaches are available to treat the symptoms. The primary approach focuses on basic skin care in the form of topical treatments with ointments, creams, or lotions to stabilize the skin barrier function and reduce its sensitivity to irritation and allergen infiltration. If this measure is insufficient in cases of more severe manifestations of the disorder or symptoms, corticosteroids are used as anti-inflammatory agents. These cortisol-like preparations are often referred to colloquially as cortisone.They are normally applied topically to areas of the skin, although they are not suitable for short-term application due to the side effects of corticosteroids, particularly even on the skin (e.g., skin atrophy, pigment disorders, and bruising). QQh / ηη / ι znz / E / YL Thus, there is a great need for active substances for the treatment of neurodermatitis and other disorders that are accompanied by fewer side effects and that can be used as an alternative to or in addition to corticosteroids. To address this problem, the present invention proposes using collagen hydrolysate as an active substance in the treatment of inflammatory skin disorders and intestinal disorders, and / or skin and intestinal disorders accompanied by inflammation. The appropriate efficacy of collagen hydrolysate has become evident, in particular, from a clinical study in which collagen hydrolysate or a placebo was administered to patients with neurodermatitis. Several beneficial effects of collagen hydrolysate have been known for a relatively long time, particularly its use in treating osteoporosis and joint pain. Positive effects of collagen hydrolysate on skin health have also been described, for example, in international patent application WO 2012 / 065782 A2. However, the efficacy of collagen hydrolysate in treating neurodermatitis and other inflammatory disorders is remarkable. Collagen hydrolysate, a byproduct of animal starting materials also used as food products, is completely harmless from a health standpoint, with no known harmful side effects. It does not require legal approval as a drug and, in particular, can be marketed and used as a food supplement. The use of collagen hydrolysate as a food supplement, as an over-the-counter (OTC) drug, or as a prescription drug (particularly in combination with other active substances) falls within the scope of the present invention. Ideally, collagen hydrolysate is administered orally. It is known that collagen hydrolysate peptides are reabsorbed in the intestine, at least to some extent, even with relatively high molecular weights up to 10,000 Da. The preferred amount of collagen hydrolysate administered as a daily dose is 2 to 15 mg, with 3 to 10 g being preferable. A positive effect in cases of neurodermatitis can be demonstrated, for example, with a daily dose of 5 g of collagen hydrolysate. In a preferred embodiment of the invention, collagen hydrolysate is used as a supportive treatment in addition to the treatment of a skin or bowel disorder with a corticosteroid. Although it is often not possible to completely discontinue the corticosteroid and still sufficiently improve symptoms, the administration of collagen hydrolysate makes it possible, at least, to reduce the amount of corticosteroid administered, which in itself results in a significant improvement in the side effects experienced by affected individuals. QQh / ηη / ι znz / E / Yl· The corticosteroid, which together with the collagen hydrolysate can be used according to the invention, is generally administered topically, specifically in the form of an ointment. Alternatively, oral administration of corticosteroids is also possible. In the latter case, according to another 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. Ideally, the corticosteroid is a glucocorticoid, for example, hydrocortisone. A large number of glucocorticoids are known, which are divided into four classes ranging from barely effective (class I) to very highly effective (class IV). A preferred embodiment of the invention relates to collagen hydrolysate for use as an active substance in the treatment of neurodermatitis. Other inflammatory skin disorders and / or skin disorders accompanied by inflammation, the treatment or supportive treatment of which is possible with collagen hydrolysate, include psoriasis, rosacea, chronic pruritus, acne, cellulitis, and dermatoporosis. The invention also comprises collagen hydrolysate for use in the treatment of inflammatory bowel disorders and / or intestinal disorders accompanied by inflammation. There are similarities between inflammatory disorders of the skin and the intestines in that the intestinal wall contains collagen fibers as structural proteins and represents a barrier between the body's interior and the external environment. Specifically, the intestinal disorders for the treatment of which collagen hydrolysate can be used within the scope of the invention are Crohn's disease and ulcerative colitis. These are chronic inflammatory bowel disorders that can be treated symptomatically with glucocorticoids. Another aspect of the invention relates to a collagen hydrolysate for use as an active substance to mitigate the side effects of corticosteroids, specifically their skin side effects, such as skin atrophy, pigment disorders, and ecchymosis. The corresponding information can also be found in the aforementioned clinical study, which will be explained in detail later. The aforementioned side effects of corticosteroids (specifically glucocorticoids) are caused in part by a reduction in collagen biosynthesis by skin cells (fibroblasts) (see, for example, Oikarinen et al., Journal of Investigative Dermatology 98 (1992) 220-225). Surprisingly, in vitro cell tests have revealed (see below) that the reduction in collagen biosynthesis can be almost completely compensated for by the presence of collagen hydrolysate. The collagen hydrolysate for use according to the invention normally has an average molecular weight of 500 to 15,000 Da, preferably 1,000 to 8,000, more preferably of 1,500 to 5,000 Da, more preferably 1,800 to 2,200 Da. In these statements, the average molecular weight is always referred to, which can be determined, in particular, by gel permeation chromatography. Ideally, collagen hydrolysate is produced by enzymatic hydrolysis of a collagen-containing starting material. Specifically, endopeptidases or exopeptidases of microbial or plant origin are used for this hydrolysis. Collagen hydrolysates within the desired molecular weight range can be produced by appropriate selection of peptidases and hydrolysis conditions. In general, the starting material containing collagen is selected from the skin or bones of vertebrates, preferably mammals or birds, and in particular, from the hides of cattle or pigs (split bovine hide or pigskin, respectively). Alternatively, the starting material containing collagen may be selected from the skin, bones, and / or scales of fish, specifically cold-water or tropical fish. Collagen hydrolysate can be produced in a one-stage method from these materials or by means of intermediate-stage gelatin; in the latter case, both type A and type B gelatin can be used. Ideally, collagen hydrolysate is produced by the sequential action of at least two endoproteases with different specificities; in particular, at least two different metalloproteases and / or serine proteases, that is, proteases that cleave the amino acid sequence of collagen molecules before or after specific amino acids. Conveniently, these metalloproteases and / or serine proteases are enzymes produced by the microorganisms *Bacichus subtilis*, *Bacichus licheniformis*, *Bacichus amyloliquefaciens*, *Aspergillus oryzae*, and *Aspergillus melleus*. By selecting appropriate endoproteases, it is possible not only to obtain a specific molecular weight distribution in the collagen hydrolysate, but also to influence the type of amino acids at the N-terminus of the peptides contained in the hydrolysate. For example, it is preferable if at least 50% of the N-terminus amino acids in the collagen hydrolysate are hydrophobic amino acids, specifically alanine, leucine, and isoleucine. As an alternative to enzymatic hydrolysis, collagen hydrolysate can be produced by recombinant gene expression within the scope of the invention. By using natural collagen sequences, specifically from cattle or pigs, and expressing them in genetically modified cells (e.g., yeast, bacteria, plant cells, specifically tobacco), products can be produced that are substantially identical to the hydrolysis products of the corresponding collagen-containing raw materials. A narrower or more precisely specified molecular weight distribution can be achieved here. The effectiveness of collagen hydrolysate in the treatment of neurodermatitis will be explained in more detail based on the clinical study described below. In addition, in vitro cell tests are described that demonstrate the effectiveness of collagen hydrolysate in mitigating the side effects of corticosteroids. In the figures: Figure 1 shows a bar graph regarding the use of a corticosteroid and a skin care ointment during the study period; Figure 2 shows a bar graph regarding the development of the skin pH value during the study period; Figure 3 shows a bar graph regarding the change in the housework / errands impairment index due to itching during the study period; QQh / ηη / ι znz / E / Yl· Figures 4A and 4B show graphs of type I collagen and proteoglycans by fibroblasts; Figures 5A and 5B show graphs of type I collagen and proteoglycans by fibroblasts; Figures 6A and 6B show graphs of type I collagen and proteoglycans by fibroblasts; bars bars bars with with with respect with respect with respect the the the biosynthesis biosynthesis biosynthesis of the of the of the 1. Clinical study 1.1 Study design The clinical study was conducted with thirty study subjects (15 men and 15 women) diagnosed with neurodermatitis, with a severity score ranging from 15 to 140 in all subjects, according to the SCORAD index. The study was a randomized, double-blind, placebo-controlled trial; that is, the subjects were randomly divided into a treatment group and a placebo group, each including 15 individuals. No study subject or treating staff member was aware of the group assignment of individual test subjects. The test subjects in the treatment group received a daily dose of 5 g of collagen hydrolysate throughout the 12-week study period. This was the product Verisol B, distributed by the applicant, which is a collagen hydrolysate produced by enzymatic hydrolysis of bovine collagen with an average molecular weight of approximately 2,000 Da. The production method for Verisol B substantially corresponds to the production method described in publication WO 2012 / 065782 A2. Test subjects in the placebo group received a daily dose of 5g of maltodextrin instead of collagen hydrolysate; the collagen hydrolysate and placebo did not differ with respect to packaging, texture, or taste. During the study, participants were free to choose whether to apply a skin care ointment (unguentum leniens) or a corticosteroid ointment (triamcinolone acetonide), as well as the amount to apply, to relieve their symptoms. The amounts of skin care ointment and corticosteroid used by each individual participant were measured throughout the study period. 1.2 Use of corticosteroid skin care ointment A notable finding of the study is that the use of both the skin care ointment and the corticosteroid ointment in the research group was significantly lower than in the placebo group. This held true both during the first third (4 weeks) of the study period and during the period from week 5 to week 12, and in each case, the difference was particularly pronounced with the use of the corticosteroid. Figure 1 shows the corresponding results in the form of a bar graph, i.e., the average use of the corticosteroid-containing ointment and the skin care ointment in grams in both groups for the respective periods. This result demonstrates the efficacy of collagen hydrolysate in the treatment of neurodermatitis. Because the amount of corticosteroid and skin care ointment can be freely selected by the test subjects as needed, the lower amount required in the research group can only be explained by the fact that some of the symptoms were directly alleviated by the orally administered collagen hydrolysate. 1.3 Measurement of the skin dH value The skin's pH value, an indicator of its barrier function, was measured in all test subjects at baseline, after four weeks, and after twelve weeks of using a skin pH meter (Courage + Khazaka electronics GmbH). Measurements were taken on the upper arm, specifically in both healthy and lesion-affected skin areas. The results are shown in Figure 2 in the form of a bar chart. In the treatment group, the pH value of the lesioned skin areas was likely to reach the pH value of healthy skin in the range of 5.3 to 5.6 over the course of the QQh / ηη / ι znz / E / YL study period. In contrast, the pH value in the placebo group, both in healthy skin and in areas of skin with lesions, increased abnormally after twelve weeks. This result leads to the conclusion that oral administration of collagen hydrolysate counteracts, at least partially, the side effects of corticosteroids on the skin. 1.4 Transdermal water loss vs. skin hydration Transdermal water loss (TEWL) measurements were taken using a Tewameter, and skin hydration was measured using a Corneometer (both from Courage + Khazaka electronics GmbH) during the study period. These parameters, similar to pH value, are indicators of skin barrier function. No significant differences were found between the treatment and placebo groups in the results for these two parameters, neither in areas of healthy skin nor in areas of skin affected by lesions. This indirectly leads to the conclusion that, despite the higher amount of corticosteroid used in the placebo group and its side effects, no deterioration of the parameters occurred, which can be attributed to the effect of the collagen hydrolysate. 1.5 Assessment of itching An assessment of itching was performed on test subjects during the study period according to a model called the 5-D Pruritus Scale. According to this model, itching is assessed both quantitatively and qualitatively (duration, severity, and tendency), with respect to body location (distribution), and also with respect to the impairment of the following life areas as a result of the itching: sleep; leisure / social activities; housework / errands; work / school. For the outcome measure of impairment of household chores / errands, a significant improvement in the index was observed in the treatment group during the study period compared to the placebo group. The corresponding results are shown in the bar graph in Figure 3 (average index values ​​at the beginning of the study, after four weeks, and after twelve weeks). This result directly reflects the efficacy of collagen hydrolysate in relieving itching. For the other assessment criterion, there were no significant differences in the various indices between the research group and the placebo group. However, considering the fact that significantly more corticosteroids were used in the placebo group, this finding also led to QQh / nn / i znz / E / Yl· indirectly to the conclusion that the effect of the corticosteroid could be replaced, at least in part, by collagen hydrolysate. 2. In vitro cell tests 2.1 Cell tests with fibroblasts Topical corticosteroid treatment is associated, in particular, with a longer treatment period and negative side effects, such as skin atrophy. A key cause of these side effects is the reduction in collagen biosynthesis by fibroblasts, as well as the biosynthesis of other key matrix proteins such as proteoglycans. Cellular assays were performed using human fibroblasts to investigate whether collagen hydrolysate in combination with glucocorticoids influences matrix protein biosynthesis. The same collagen hydrolysate (Verisol B) was used for these assays as in the clinical study described above. Primary human fibroblasts were cultured in HAM F12 medium supplemented with 10% fetal bovine serum, 20 U / ml penicillin streptomycin, 50 pg 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 either 0.5 mg / ml collagen hydrolysate, 0.05 mg / ml a glucocorticoid (dexamethasone), or a combination of both. A control batch was also cultured in the culture medium without any additions. After an additional seven-day culture, the amount of extracellular matrix proteins synthesized by the fibroblasts was determined, with type I collagen determined by the Sircol Soluble Collagen Assay, and proteoglycans by the Blyscan Glycosaminoglycan Assay (biocolor Ltd, Great Britain), in each case according to the manufacturer's instructions. The results are shown in the bar graphs in Figures 4A-4B, where Figure 4A relates to type I collagen biosynthesis, and Figure 4B relates to proteoglycan biosynthesis. In each case, the average values ​​from eight tests are specified in relation to the control batch. In both cases, it can be observed that collagen and proteoglycan biosynthesis are significantly reduced by glucocorticoids, as expected. However, this reduction can be completely offset by supplementation with collagen hydrolysate. Collagen hydrolysate itself has a moderately stimulatory effect on biosynthesis. αοίτ / ηη / ίζηζ / Ε / γι These results are further indication 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 administering collagen hydrolysate. 2.2 Cellular tests with chondrocytes The same cell tests were also performed with human chondrocytes (cartilage cells). Cell culture and the determination of type I collagen and biproteoglycans were carried out as in the tests described above with fibroblasts, with the difference that a collagen hydrolysate from bovine bone gelatin with an average molecular weight of approximately 4,000 Da was used. The results are shown in the bar graphs in Figures 5A–5B, where Figure 5A shows type I collagen biosynthesis, and Figure 5B shows proteoglycan biosynthesis. In each case, the average values ​​for these three tests are shown. It can also be observed here that the reduction in biosynthesis caused by the glucocorticoid can be substantially offset by the addition of collagen hydrolysate. 2.3 Cellular tests with osteoblasts Corresponding tests were also performed with human osteoblastomas (bone cells); the tests were conducted and type I collagen and proteoglycans were determined in the same manner as for the chondrocyte tests (including collagen hydrolysate with 4,000 Da). The bar graph in Figures 6A-6B shows that, also in the case of osteoblasts, the reduction in type I collagen biosynthesis (Figure 6A) and proteoglycans (Figure 6B) by the glucocorticoid can be substantially compensated for by the addition of collagen hydrolysate. The results with chondrocytes and osteoblasts show that the advantageous effects of collagen hydrolysate in combination with corticosteroids are not limited to the skin, and that relief from side effects can also be expected in other types of tissue.

Claims

1. A collagen hydrolysate for use as an active substance in the treatment of inflammatory skin disorders and intestinal disorders and / or skin and intestinal disorders accompanied by inflammation.

2. The collagen hydrolysate for use as claimed in claim 1, wherein the collagen hydrolysate is administered orally, preferably in an oral dose of 2 to 15 g, more preferably 3 to 10 g.

3. The collagen hydrolysate for use as claimed in claim 1 or 2, wherein the collagen hydrolysate is used for supportive treatment in addition to treatment of a skin disorder or bowel disorder with a corticosteroid.

4. The collagen hydrolysate for use as claimed in claim 3, wherein the administration of the collagen hydrolysate allows a reduction in the amount of corticosteroid administered.

5. The collagen hydrolysate for use as claimed in any of claims 2 to 4, wherein the corticosteroid is administered topically, specifically in the form of an ointment.

6. The collagen hydrolysate for use as claimed in any of claims 2 to 4, wherein the corticosteroid is administered orally, specifically in the form of a composition containing the collagen hydrolysate and the corticosteroid.

7. The collagen hydrolysate for use as claimed in any of claims 2 to 6, wherein the corticosteroid is a glucocorticoid, for example, hydrocortisone.

8. The collagen hydrolysate according to any of the preceding claims, further characterized in that the skin disorder is neurodermatitis.

9. The collagen hydrolysate for use as claimed in any of claims 1 to 7, wherein the skin disorder is selected from psoriasis, rosacea, chronic pruritus, acne, cellulitis, and dermatoporosis.

10. The collagen hydrolysate for use as claimed in any of claims 1 to 7, wherein the intestinal disorder is selected from Crohn's disease and ulcerative colitis.

11. A collagen hydrolysate for use as an active substance to mitigate the side effects of corticosteroids, specifically their side effects on the skin, such as skin atrophy, pigment disorders, and ecchymosis.

12. The collagen hydrolysate for use as claimed in any of the preceding claims, wherein the collagen hydrolysate has an average molecular weight of 500 to 15,000 Da, preferably 1,000 to 8,000, more preferably 1,500 to 5,000 Da, more preferably 1,800 to 2,200 Da.

13. The collagen hydrolysate for use as claimed in any of the preceding claims, wherein the collagen hydrolysate is produced by enzymatic hydrolysis of a starting material containing collagen.

14. The collagen hydrolysate for use as claimed in claim 13, wherein the starting material containing collagen is selected from the skin or bones of vertebrates, preferably mammals, birds or fish, specifically from the skin of cattle or pigs.

15. The collagen hydrolysate for use as claimed in claim 13 or 14, wherein the collagen hydrolysate is produced by the successive action of at least two endoproteases having different specificities, specifically, at least two metalloproteases and / or serine proteases.

16. The collagen hydrolysate for use as claimed in claim 15, wherein the metalloproteases and / or serine proteases are selected from enzymes of the microorganisms Bacillus subtilis, Bacillus chemiformis, Bacillus amyloliquefaciens, Aspergillus oryzae, and Aspergillus meieus.

7. The collagen hydrolysate for use as claimed in any of the preceding claims, wherein at least 50% of the N-terminal amino acids of the collagen hydrolysate are hydrophobic amino acids, specifically alanine, leucine, and isoleucine.

18. The collagen hydrolysate for use as claimed in any of claims 1 to 12, wherein the collagen hydrolysate is produced by recombinant gene expression.