Casein hydrolysate for treating crohn's disease

Abstract

The invention relates to a casein hydrolysate for the treatment of Crohn's disease. The composition of the casein hydrolysate is determined by the use of a proline- specific endoprotease for the hydrolysis of casein and is characterized in that the molar fraction of peptides of said hydrolysate carrying a carboxy terminal proline, expressed in %, is more than two times the molar fraction of proline, expressed in %, in the casein substrate used to generate the hydrolysate.

Description

[0001] CASEIN HYDROLYSATE FOR TREATING CROHN’S DISEASE

[0002] TECHNICAL FIELD

[0003] The present invention relates to the treatment of Crohn’s disease.

[0004] BACKGROUND ART

[0005] Crohn’s disease is a type of inflammatory bowel disease characterized by mucosal ulceration and inflammation, which may occur in any part of the gastrointestinal tract, but most commonly affects the distal small intestine.

[0006] The etiology of Crohn’s disease is not completely understood, but probably involves a combination of genetic, immunological and environmental factors. It is believed that its onset is triggered by environmental factors that perturb the mucosal barrier, alter the healthy balance of the gut microbiota, and abnormally stimulate gut immune responses (Boyapati et al., Pathogenesis of Crohn’s disease, 2015, FlOOOPrime Rep. ,7:44).

[0007] Symptoms of Crohn’s disease include diarrhea, rectal bleeding, abdominal pain, fatigue and weight loss. Furthermore, Crohn’s disease may lead to other complications, such as bowel obstruction, ulcers, fistulas, anal fissure, malnutrition or colorectal cancer.

[0008] Crohn’s disease is characterized by its progressive and destructive course, unpredictable nature, and high morbidity rate, which can result in long-lasting physical, emotional, and psychological effects on patients.

[0009] At present, there is no cure for Crohn’s disease, and the treatment options currently available are aimed to the reduction of inflammation and control of symptoms, improvement of quality of life and to minimize complications.

[0010] Common treatments for Crohn’s disease include steroids, 5-aminosalicylates (5-ASA), antibiotics, and immunosuppressants, which can be used in outbreaks of the disease. Newer treatments include anti-TNF biologies, such as infliximab, adalimumab or certolizumab pegol; or other more recent biologies, such as vedolizumab and ustekinumab (Kumar et al., A review of the therapeutic management of Crohn’s disease, Ther. Adv. Gastroenterol., 2022, 15, 1-19).

[0011] Despite the treatments available, surgery is ultimately frequently needed, especially for the treatment of complications, such as perforation, abscesses, stenotic areas, fistulas or malignancy.

[0012] Furthermore, the therapeutic options available so far are not without drawbacks. The conventional therapeutic management, with steroids, 5-ASA or antibiotics, are of limited efficacy and entail frequent side effects. Anti-TNF medications have also problems, such as high rates of primary and secondary non-responders, and the long-term risk for complications. And despite the improvements brought by the development of new biologies, there is still a high rate of surgery and post-operative recurrence (Kumar et al., op.cit.).

[0013] Therefore, despite the therapeutic options available so far, there remains a need for new treatment options for Crohn’s disease, which are effective for treating its different symptoms and manifestations and also for preventing future recurrences and complications, and where the side effects of the treatment are minimized.

[0014] SUMMARY OF THE INVENTION

[0015] The object of the present invention is a casein hydrolysate for use in the treatment of Crohn’s disease.

[0016] DETAILED DESCRIPTION OF THE INVENTION

[0017] The object of the present invention is a casein hydrolysate for use in the treatment and / or prevention of Crohn’s disease, wherein the hydrolysate comprises peptides wherein the molar fraction of peptides carrying a carboxy terminal proline, expressed in %, is more than two times the molar fraction of proline, expressed in %, in the casein substrate used to generate the hydrolysate.

[0018] The authors of the present invention have developed a new therapy for the treatment of Crohn’s disease, based on the use of a specific casein hydrolysate, which, surprisingly, provides outstanding efficacy in the elimination of the symptoms associated to Crohn’s disease, even leading to complete remission of the disease, as well as effective prevention of its derived complications.

[0019] Definitions

[0020] Along the present description, as well as in the claims, singular expressions, generally preceded by the articles “a”, “an” or “the”, are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0021] The terms “about” or “approximately” referred to amounts, as used herein, are meant to include the exact amount and also a certain deviation around the stated amount, namely of ±5%.

[0022] The numerical ranges disclosed herein, generally expressed as “from ... to ...” or as “between ... and ...”, are meant to include any number falling within the ranges and also the lower and upper limits.

[0023] Along the present description, as well as in the claims, the percentages are generally by weight (%w, % w / w), unless the context clearly indicates otherwise. Thus, for example, the percentage of the various components of the described formulations refers to their proportion by weight relative to the total weight of the composition. However, the percentages that define the peptide composition of the casein hydrolysate clearly refer to the molar fraction, and in this specific case it is clearly understood that they are not percentages by weight.

[0024] Casein hydrolysate

[0025] The casein hydrolysate is a specific hydrolysate that is characterized in that the molar fraction of the peptides of said hydrolysate carrying a carboxy terminal proline, expressed in %, is more than two times the molar fraction of proline, expressed in %, in the casein substrate used to generate the hydrolysate.

[0026] As is well known in the art, the term casein encompasses a group of phosphoproteins present in the milk, which represents approximately a 3% of the bovine milk. The main components of casein are alpha-, beta-, gamma- and kappa-caseins, among which beta casein is the major fraction of bovine milk.

[0027] Casein used as substrate for the hydrolysis is preferably casein from bovine milk, more preferably beta-casein from bovine milk.

[0028] Although several bovine beta-casein variants have been described, they only show minor amino acid differences (Cieslihska et al. Int J Mol Sci. 2022 Dec 9;23(24):15637), and all are suitable to be used as substrate for preparing the hydrolysate according to the present invention. For example, bovine beta-casein A1 or bovine betacasein A2 can be used, or a mixture of both.

[0029] The casein hydrolysate for use according to the present invention, is a hydrolysate whose composition is determined by the use of a proline specific endoprotease in the hydrolysis of the casein, so that the hydrolysate has a composition characterized by a high content of peptides with a proline at the carboxy-terminal end.

[0030] In the use of the invention, the molar fraction of the peptides in such hydrolysate carrying a carboxy terminal proline, expressed in %, is more than two times the molar fraction of proline, expressed in %, in the casein substrate used to generate hydrolysate.

[0031] The characteristics of this hydrolysate and of the proline specific endopeptidase used in its preparation are described in the international patent application WO-A-02 / 45524 or in the international patent application WO-A-02 / 45523. In that document it is also disclosed the method for determining the molar fraction of the peptides carrying a carboxy terminal proline, expressed in %, as well as the method for determining the molar fraction of proline, expressed in %, in the casein substrate used to generate the hydrolysate. Preferably, the molar fraction of the peptides of this hydrolysate carrying a carboxy terminal proline, expressed in %, is at least three times the molar fraction of proline, expressed in %, in the casein substrate used to generate the hydrolysate.

[0032] Preferably, in the casein hydrolysate according to the use of the present invention, the average length of the peptides in the hydrolysate is comprised between 3 and 9 amino acids.

[0033] Preferably, the molar fraction of peptides carrying a carboxy terminal proline in the casein hydrolysate is at least 25%, and still more preferably it ranges between 30% and 70%.

[0034] Within the context of the present invention, when discussing the molar fraction of peptides, peptides are understood to be those whose molecular mass ranges between 400 and 2000 Dalton, which can be determined, for example, using the liquid chromatography / mass spectrometry (LC / MS) method described in the Materials and Methods section of the international patent application WO-A-02 / 45524 cited above.

[0035] In general, the casein hydrolysate according to the use of the present invention is a hydrolysate where at least the 50% of the casein substrate is hydrolysed. Preferably at least a 10% of the casein substrate is converted into peptides with a molecular mass between 400 and 2000 Dalton, more preferably, between 20% and 90%, and still more preferably between 30% and 80% of the casein substrate is converted into such peptides.

[0036] In a preferred embodiment the casein hydrolysate has the following amino acid composition: between 54-64 of lysine, between 22-30 of methionine, between 25-33 of threonine, between 18-26 of histidine, between 26-34 of arginine, between 46-56 of valine, between 34-42 of isoleucine, between 70-80 of leucine, between 33-41 of phenylalanine, between 45-55 of the sum of aspartic acid plus asparagine, between 160-180 of the sum of glutamine plus glutamic acid, between 20-26 of alanine, between 80-90 of proline, between 40-48 of tyrosine, between 37-45 of serine, between 11-17 of glycine, between 0.8-1.2 of cysteine, and between 6-10 of tryptophan; wherein the amounts are expressed in grams of each amino acid per kilogram of hydrolysate.

[0037] In a more preferred embodiment the casein hydrolysate has the following amino acid composition: between 57-61 of lysine, between 24-28 of methionine, between 27-31 of threonine, between 20-24 of histidine, between 28-32 of arginine, between 49-53 of valine, between 36-40 of isoleucine, between 73-77 of leucine, between 35-39 of phenylalanine, between 48-52 of the sum of aspartic acid plus asparagine, between 166- 176 of the sum of glutamine plus glutamic acid, between 21-25 of alanine, between 82-88 of proline, between 42-46 of tyrosine, between 39-43 of serine, between 13-15 of glycine, between 0.8-1.2 of cysteine and between 7-9 of tryptophan; wherein the amounts are expressed in grams of each amino acid per kilogram of hydrolysate.

[0038] In a particularly preferred embodiment the casein hydrolysate has approximately the following amino acid composition: 59 of lysine, 26 of methionine, 29 of threonine, 22 of histidine, 30 of arginine, 51 of valine, 38 of isoleucine, 75 of leucine, 37 of phenylalanine, 50 of the sum of aspartic acid plus asparagine, 171 of the sum of glutamine plus glutamic acid, 23 of alanine, 85 of proline, 44 of tyrosine, 41 of serine, 14 of glycine, 1 of cysteine, and 8 of tryptophan; wherein the amounts are expressed in grams of each amino acid per kilogram of hydrolysate.

[0039] A casein hydrolysate according to the specified characteristics is available under the trademark PeptoPro® (DSM). According to its data sheet, this product has applications in food and beverages to enrich their protein content.

[0040] Furthermore, a casein hydrolysate according to these characteristics is also commercially available under the name HuPaVir®, for the treatment and prevention of HPV infections, including the treatment of condyloma.

[0041] Treatment of Crohn’s disease

[0042] The casein hydrolysate according to the present invention is for use in the treatment and / or prevention of Crohn’s disease.

[0043] As shown in Examples 2 to 4, the authors of the present invention found that, surprisingly, the casein hydrolysate according to the characteristics specified above is outstandingly effective for treating the symptoms of Crohn’s disease, achieving the remission or reduction of the disease outbreaks, and for the successful prevention of its complications.

[0044] In the context of the present invention, the term "treatment" of Crohn’s disease refers to the use of the hydrolysate for the purpose of curing the symptoms and signs of this disease in patients diagnosed with Crohn’s disease. By cure is meant either the complete elimination of signs and symptoms, or their alleviation, mitigation or improvement. The signs and symptoms typically found in patients suffering from Crohn’s disease include persistent diarrhoea, rectal bleeding, blood in stool, nausea, vomiting, abdominal pain, abdominal cramping, loss of appetite and weight loss and fatigue, among others.

[0045] The term “prevention” of Crohn’s disease, as used herein, refers to the prophylactic use of the casein hydrolysate, with the purpose for preventing or delaying the onset of the disease, including the maintenance of long-term remission by avoiding appearance of new outbreaks of the disease after total or partial remission. The preventive treatment is also aimed to avoid or minimize the risk of possible complications in patients diagnosed with Crohn’s disease, said complications typically including bowel obstruction, fistulas, strictures and abscesses, as well as osteoporosis, colorectal cancer, arthritis, uveitis, scleritis, episcleritis, erythema nodosum, pyoderma gangrenosum, or psoriasis, among others. Another aspect of the preventive use of the casein hydrolysate is to avoid the need for surgical intervention. Another aspect of the preventive use of the casein hydrolysate is to maintain adequate nutrition and improve quality of life.

[0046] A particular aspect of the invention is the casein hydrolysate according to the invention for use in the prevention of colorectal cancer in patents with Crohn’s disease.

[0047] As is well-known in the art, the diagnosis of Crohn’s disease involves clinical assessment, combined with endoscopic, histologic, radiologic, and / or biochemical testing (Veauthier et al., Crohn’s disease: Diagnosis and Management, Am. Fam. Physicial 2018, 98(11), 661-669; or R. Arumugam and R. Sinha R, Editors, Crohn’s disease. Current concepts, Springer International Publishing Switzerland, 2015).

[0048] Examples 2 to 3 show the efficacy of the casein hydrolysate of the present invention for the treatment of different groups of patients with Crohn’s disease. Furthermore, Example 4 shows its efficacy for colorectal cancer prevention in patients diagnosed with Crohn’s disease, as shown by the serum levels of two biochemical markers of colorectal cancer, namely, carcinoembryonic antigen (CEA) and carbohydrate antigen (CA 19.9).

[0049] As the product used in the present invention is completely harmless, derived from the hydrolysis of casein from bovine milk and commonly used as a dietetic supplement, the use according to the present invention has the advantage that it is possible to maintain a long preventive treatment, without the risk of experiencing the undesirable side effects associated to other pharmaceutical treatments, so that the patients taking preventative doses are able to remain without any outbreak of the disease for very long periods, as shown in the examples. The safety of the present treatment is particularly beneficial for the prevention serious complications of Crohn’s disease, such as colorectal cancer.

[0050] Optionally, the casein hydrolysate according to the present invention may be used in combination with another drug for the treatment of Crohn’s disease. The drug to be combined with the casein hydrolysate is meant to be a drug effective against Crohn's disease, as known in the art, for example, 5-aminosalicylates (such as sulfasalazine), antibiotics (such as metronidazole, ciprofloxacin or rifaximin), corticosteroids (such as prednisone, prednisolone, methylprednisolone or budesonide), immunosuppressants (such as azathioprine, 6-mercaptopurine, methotrexate or cyclosporine), tumor necrosis factor (TNF) alpha antagonists (such as infliximab, adalimumab, certolizumab pegol or golimumab), anti-integrin therapies (such as natalizumab or vedolizumab), or interleukin (I L)-12 and IL-23 inhibitors (such as ustekinumab), among others. Therefore, another aspect of the invention is the casein hydrolysate as above defined in combination with at least one drug for use in the treatment and / or prevention of Crohn’s disease.

[0051] Another aspect of the invention is a method for the treatment and / or prevention of Crohn’s disease, comprising administering a therapeutically effective dose of the casein hydrolysate of the invention to a subject in need thereof.

[0052] The casein hydrolysate according to the use of the present invention is administered orally.

[0053] Typically, the casein hydrolysate is administered according to a daily oral dose comprised between 3 and 40 g, preferably comprised between 5 and 30 g. The exact dose will be easily adjusted by the skilled practitioner according to each particular case, depending on whether it is for the treatment of the active disease or for maintenance / preventive treatment, and also depending on the type and severity of the symptom to be treated.

[0054] In general, for the treatment of mild symptoms or for preventive / maintenance therapy, a daily dose comprised between 5 and 15 g can be used; while for treatment of more severe symptoms, a daily dose comprised between 10 and 30 g can be used.

[0055] In an embodiment of the invention, a unit oral dose comprising between 4 and 8 g of the casein hydrolysate is used, preferably comprised between 5.5 and 6.5 g, more preferably about 6 g. This unit dose is typically administered from 1 to 5 times daily, and more preferably from 1 to 3 times daily, i.e., according to a once daily, twice daily or three times daily regimen. For example, a once daily regimen can be typically used for preventive / maintenance therapy or for the treatment of less severe symptoms, while twice or three times daily regimen can be used for the treatment of more severe symptoms.

[0056] The casein hydrolysate according to the use of the present invention is typically available as a powder substance, and can be administered either directly or in combination with at least one pharmaceutically acceptable excipient and / or carrier, in the form of a pharmaceutical composition.

[0057] Another aspect of the invention is a casein hydrolysate for use in the treatment and / or prevention of Crohn’s disease, characterized in that the hydrolysate is administered in the form of a pharmaceutical composition comprising this casein hydrolysate and at least one pharmaceutically acceptable excipient and / or carrier, wherein this hydrolysate comprises peptides wherein the molar fraction of peptides carrying a carboxy terminal proline, expressed in %, is more than two times the molar fraction of proline, expressed in %, in the casein substrate used to generate the hydrolysate. The pharmaceutical composition can be prepared using methods that are well known to the skilled in the art such as those contained in handbooks of pharmaceutical technology, such as the book Remington, The Science and Practice of Pharmacy, 20th edition, Lippincott, Williams & Wilkins, Philadelphia, 2000 [ISBN: 0-683-306472],

[0058] In an embodiment of the invention, the pharmaceutical composition is a composition suitable for oral administration.

[0059] Any pharmaceutical form suitable for oral administration is included, preferably solid compositions in powder or granulate form, but also liquid compositions, in solution, suspension or syrup form, for example.

[0060] Preferably, the pharmaceutical composition for oral administration is in powder or granulate form. More preferably, it is in powder form.

[0061] The pharmaceutically acceptable excipients that can be used for preparing pharmaceutical compositions in solid form are well known to those skilled in the art and include, for example, diluents such as calcium carbonate, sodium carbonate, magnesium carbonate, magnesium oxide, calcium sulfate, calcium phosphate, sodium chloride, microcrystalline or powdered cellulose, cellulose acetate, ethyl cellulose, dextrates, dextrins, dextrose, lactose, lactitol, fructose, sorbitol, sucrose, maltodextrins, maltose, glyceryl palmitostearate, kaolin, polymethacrylates, pregelatinized starch or starch, among others, and mixtures thereof; lubricants, such as calcium stearate, magnesium stearate, talc, stearic acid, glyceryl behenate, glyceryl palmitostearate, sodium benzoate, sodium lauryl sulfate, sodium stearyl fumarate, or hydrogenated castor oil, among others, and mixtures thereof; disintegrants such as alginic acid, sodium croscarmellose, crospovidone, sodium starch glycolate, starch, low-substituted hydroxypropylcellulose, among others, and mixtures thereof; binding agents such as sodium carboxymethylcellulose, cellulose acetate phthalate, dextrates, dextrin, ethylcellulose, guar gum, maltodextrin, methylcellulose, microcrystalline cellulose, povidone, pregelatinized starch, stearic acid, or sucrose, among others, and mixtures thereof; anticaking agents such as tribasic calcium phosphate, calcium silicate, colloidal silica, magnesium silicate, magnesium trisilicate, or talc, among others, and mixtures thereof; thickening agents such as colloidal silica, dextrin, ethylcellulose, hydroxyethyl cellulose, hydroxyethylmethyl cellulose, hypromellose, polyethylene glycol, trehalose, xanthan gum, among others, and mixtures thereof; suspending agents such as xanthan gum, guar gum, alginic acid, bentonite, carbomers, sodium or calcium carboxymethylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, hydroxypropyl alginate, microcrystalline or powdered cellulose, anhydrous colloidal silica, dextrins, gelatins, kaolin, magnesium aluminum silicate, maltitol, povidone, sorbitan esters, or tragacanth, among others, and mixtures thereof; stabilizing agents such as guar gum, xanthan gum, alginic acid, ascorbic acid, calcium stearate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, ethylcellulose, lecithin, monoethanolamine, potassium chloride, povidone, sorbitol, or xylitol, among others, and mixtures thereof; flavoring agents such as maltol, vanillin, ethyl vanillin, menthol, citric acid, fumaric acid, ethyl maltol, tartaric acid, peppermint, artificial or natural fruit aromas, among others, and mixtures thereof; sweetening agents such as sorbitol, maltitol, mannitol, dextrose, maltose, xylitol, saccharin, sucrose, sucralose, aspartame, acesulfame potassium, or trehalose, among others, and mixtures thereof; coloring agents such as curcumin, lactoflavin, iron oxides (red, yellow or black), caramel, lactoflavin phosphate, cochineal red, titanium dioxide, or carotenes, among others, and mixtures thereof; or mixtures thereof.

[0062] Some of the excipients and carriers suitable to be used in the liquid formulations, in the form of solutions, or suspensions are, for example, solvents such as water, alcohol, almond oil, castor oil, glycerin, among others; buffering agents such as diethanolamine, dibasic sodium phosphate, monobasic sodium phosphate, potassium citrate, sodium bicarbonate, sodium citrate dihydrate, among others, and mixtures thereof; viscosity modifiers such as alginic acid, bentonite, carbomers, carrageenan, gelatin, glycerin, hydroxyethyl cellulose, hydroxyethylmethyl cellulose, hydroxypropyl cellulose, maltodextrin, polyvinyl alcohol, sodium alginate, tragacanth, arabic gum, or xanthan gum, among others and mixtures thereof; emulsifying agents such as calcium stearate, cetyl alcohol, ethylene glycol palmitostearate, glyceryl monostearate, lecithin, oleic acid, poloxamers, sodium lauryl sulfate, sorbitan esters, polyoxyethylene castor oil derivatives, or emulsifying wax, among others, and mixtures thereof; suspending agents such as xanthan gum, guar gum, alginic acid, bentonite, carbomers, sodium or calcium carboxymethylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, hydroxypropyl alginate, microcrystalline or powdered cellulose, anhydrous colloidal silica, dextrins, gelatins, kaolin, magnesium aluminum silicate, maltitol, povidone, sorbitan esters, or tragacanth, among others, and mixtures thereof; flocculating agents such as sodium acetate, sodium phosphate, sodium citrate, sodium lauryl sulfate, starch, alginates, tragacanth, or carbomers, among others, and mixtures thereof; wetting agents as benzalkonium chloride, sodium docusate, sodium lauryl sulfate, sorbitan esters, polyoxyethylene stearates or polyoxyethylene sorbitan fatty acid esters, among others, and mixtures thereof; preservatives such as benzalkonium chloride, benzyl alcohol, bronopol, parabens, sodium benzoate, sodium propionate, sorbic acid or thimerosal, among others, and mixtures thereof; flavoring agents as maltol, vanillin, ethyl vanillin, menthol, citric acid, fumaric acid, ethyl maltol, tartaric acid, peppermint, artificial or natural fruit aromas, among others, and mixtures thereof; sweetening agents such as sorbitol, maltitol, mannitol, dextrose, maltose, xylitol, saccharin, sucrose, sucralose, aspartame, acesulfame potassium or trehalose, among others, and mixtures thereof; coloring agents such as curcumin, lactoflavin, iron oxides (red, yellow or black), caramel, lactoflavin phosphate, cochineal red, titanium dioxide, or carotenes, among others, and mixtures thereof; or mixtures thereof.

[0063] The excipients suitable to be used in the pharmaceutical compositions of the present invention are well known to those skilled in pharmaceutical technology and are described, for example, in the book R.C. Rowe, P.J. Sheskey and P.J. Weller, Handbook of Pharmaceutical Excipients, Fourth Edition, Pharmaceutical Press, 2003.

[0064] In a preferred embodiment, the pharmaceutical composition is in the form of powder or granulate for oral use.

[0065] The powder is usually prepared by mixing the casein hydrolysate in powder form with at least one pharmaceutically acceptable excipient.

[0066] The granulate consists of powder particles that have been aggregated to form larger particles, and it is prepared according to procedures which are well known to those skilled in the art, such as dry granulation or wet granulation.

[0067] The compositions in the form of powder or granulate are usually taken after dissolution or dispersion in water or another liquid.

[0068] In an embodiment of the invention, the powder or granulate composition is presented in a bulk container as, for example, in a glass or plastic container with a wide opening, so that the required dose for each administration is taken as needed, preferably with the help of a measurement device or dispenser to measure the dose to be administered.

[0069] In another embodiment of the invention, the powder or granulate composition is presented in the form of monodose sachets, containing the unit dose suitable for oral administration. Those sachets can be made of paper or either of aluminum or plastic laminates. Preferably, the oral unit dose comprises between 4 and 8 g of the casein hydrolysate, more preferably between 5 and 7 g, and still more preferably about 6 g of the hydrolysate.

[0070] In a particularly preferred embodiment of the invention, the composition for oral use is in powder form. More preferably, the composition in powder form comprises the casein hydrolysate, and a pharmaceutically acceptable excipient selected from the group consisting of a sweetener, a flavoring agent, a coloring agent, and mixtures thereof.

[0071] Preferably, the composition in powder form contains a quantity of the casein hydrolysate comprised between 70% and 99%, and more preferably comprised between 80% and 95%, expressed as weight ratio relative to the total weight of the composition.

[0072] The present invention is defined by the following embodiments: 1.- A casein hydrolysate for use in the treatment and / or prevention of Crohn’s disease, wherein the hydrolysate comprises peptides wherein the molar fraction of peptides carrying a carboxy terminal proline, expressed in %, is more than two times the molar fraction of proline, expressed in %, in the casein substrate used to generate the hydrolysate.

[0073] 2.- A casein hydrolysate for use according to embodiment 1 , wherein the molar fraction of the peptides in the hydrolysate carrying a carboxy terminal proline, expressed in %, is at least three times the molar fraction of proline, expressed in %, in the casein substrate used to generate the hydrolysate.

[0074] 3.- A casein hydrolysate for use according to embodiments 1 or 2, wherein the average length of the peptides in the casein hydrolysate is comprised between 3 and 9 amino acids.

[0075] 4.- A casein hydrolysate for use according to embodiment 1 , wherein the molar fraction of peptides carrying a carboxy terminal proline in the casein hydrolysate is at least 25%.

[0076] 5.- A casein hydrolysate for use according to embodiment 4, wherein the molar fraction of peptides carrying a carboxy terminal proline in the casein hydrolysate is between 30% and 70%.

[0077] 6.- A casein hydrolysate for use according to any one of embodiments 1 to 5, wherein the casein hydrolysate has the following amino acid composition: between 54-64 of lysine, between 22-30 of methionine, between 25-33 of threonine, between 18-26 of histidine, between 26-34 of arginine, between 46-56 of valine, between 34-42 of isoleucine, between 70-80 of leucine, between 33-41 of phenylalanine, between 45-55 of the sum of aspartic acid plus asparagine, between 160-180 of the sum of glutamine plus glutamic acid, between 20-26 of alanine, between 80-90 of proline, between 40-48 of tyrosine, between 37-45 of serine, between 11-17 of glycine, between 0.8-1.2 of cysteine, and between 6-10 of tryptophan; wherein the amounts are expressed in grams of each amino acid per kilogram of hydrolysate.

[0078] 7.- A casein hydrolysate for use according to any one of embodiments 1 to 6, wherein it is for use in the prevention of colorectal cancer in patients with Crohn’s disease.

[0079] 8.- A casein hydrolysate for use according to any one of embodiments 1 to 7, wherein it is for use in combination with at least one drug for use in the treatment and / or prevention of Crohn’s disease. 9.- A casein hydrolysate for use according to any one of embodiments 1 to 8, wherein the casein hydrolysate is for oral administration.

[0080] 10.- A casein hydrolysate for use according to embodiment 9, wherein the casein hydrolysate is for administration according to a daily dose comprised between 3 and 40 g, preferably comprised between 5 and 30 g.

[0081] 11.- A casein hydrolysate for use according to embodiment 10, wherein the casein hydrolysate is for administration according to a daily dose comprised between 5 and 15 g.

[0082] 12.- A casein hydrolysate for use according to embodiment 10, wherein the casein hydrolysate is for administration according to a daily dose comprised between 10 and 30 g.

[0083] 13.- A casein hydrolysate for use according to embodiment 10, wherein a unit oral dose of between 4 and 8 g of the casein hydrolysate is used, which is administered from 1 to 5 times daily.

[0084] 14.- A casein hydrolysate for use according to any one of embodiments 1 to 13, wherein the casein hydrolysate is in the form of a pharmaceutical composition comprising a therapeutically effective dose of the hydrolysate and at least one pharmaceutically acceptable excipient and / or carrier.

[0085] 15.- A casein hydrolysate for use according to embodiment 14, wherein the pharmaceutical composition is a composition suitable for oral administration.

[0086] 16.- A casein hydrolysate according to embodiment 15, wherein the pharmaceutical composition is in powder or granulate form.

[0087] 17.- A casein hydrolysate for use according to embodiment 16, wherein the composition is dosed in monodose sachets containing between 4 and 8 g of the casein hydrolysate. Preparation of a composition in powder form

[0088] A composition in powder form was prepared using the following components:

[0089] All ingredients were thoroughly mixed until obtaining a homogeneous mixture and the mixture was introduced into a monodose sachet.

[0090] Each monodose sachet contained 6 g of casein hydrolysate.

[0091] Example 2.- Efficacy in symptom remission in Crohn’s disease patients

[0092] Several patients diagnosed with Crohn’s disease, were grouped according to their different symptoms and treated with the product of Example 1 (6 g per dose).

[0093] The results of the treatment are summarized in the following table. The first column (G) refers to the group number (1 to 6), the second column (n) shows the number of patients in each group, the third column (dose) shows the number of daily doses (of 6g each), the fourth column (t) shows the duration of the treatment, wherein “m” stands for months and “w” stands for weeks, the fifth column (“initial condition”) describes the symptoms of the patients before the treatment, and the last column (“assessment after treatment”) describes the evolution of the symptoms after the treatment: Efficacy in symptom remission in Crohn’s disease patients

[0094] 12 patients diagnosed with Crohn's disease (6 men and 6 women), aged between 30 and 50 years, were treated with 2 monodose sachets daily of the powder product of Example 1 , each containing 6 g of the casein hydrolysate, during 30 days.

[0095] In all cases, a reduction in inflammation, normalization of the consistency and frequency of stools, and weight gain were observed.

[0096] After symptomatic and physiological improvement, patients continued the treatment with 1 mono-dose sachet of the product daily, for 18 additional months, and with this dosage no relapses in inflammation, diarrhea, or associated weight loss were observed. Efficacy for colorectal cancer prevention in patients with Crohn’s disease

[0097] The effectiveness of the casein hydrolysate in preventing the appearance of colorectal cancer in patients with Crohn’s disease was assessed through the measurement of two biochemical markers of colorectal cancer, namely, CEA (carcinoembryonic antigen) and CA 19.9 (carbohydrate antigen). The study was carried out with 4 patients (2 women and 2 men), aged from 35 to 55 years old.

[0098] For all patients, the serum concentrations of these markers were determined before starting the treatment. The patients were administered 1 dose (6 g) daily of the product of Example 1. After 48 months of treatment no significant increase was detected in the serum concentration of the markers, no clinical signs were detected, and the symptoms associated with Crohn's disease were reduced in quantity and intensity.

Claims

CLAIMS1.- A casein hydrolysate for use in the treatment and / or prevention of Crohn’s disease, wherein the hydrolysate comprises peptides wherein the molar fraction of peptides carrying a carboxy terminal proline, expressed in %, is at least three times the molar fraction of proline, expressed in %, in the casein substrate used to generate the hydrolysate, wherein the casein substrate used to generate the hydrolysate is bovine beta-casein.2.- A casein hydrolysate for use according to claim 1 , wherein the average length of the peptides in the casein hydrolysate is comprised between 3 and 9 amino acids.3.- A casein hydrolysate for use according to claim 1 , wherein the molar fraction of peptides carrying a carboxy terminal proline in the casein hydrolysate is at least 25%.4.- A casein hydrolysate for use according to claim 3, wherein the molar fraction of peptides carrying a carboxy terminal proline in the casein hydrolysate is between 30% and 70%.5.- A casein hydrolysate for use according to any one of claims 1 to 4, wherein the casein hydrolysate has the following amino acid composition: between 54-64 of lysine, between 22-30 of methionine, between 25-33 of threonine, between 18-26 of histidine, between 26- 34 of arginine, between 46-56 of valine, between 34-42 of isoleucine, between 70-80 of leucine, between 33-41 of phenylalanine, between 45-55 of the sum of aspartic acid plus asparagine, between 160-180 of the sum of glutamine plus glutamic acid, between 20-26 of alanine, between 80-90 of proline, between 40-48 of tyrosine, between 37-45 of serine, between 11-17 of glycine, between 0.8-1.2 of cysteine, and between 6-10 of tryptophan; wherein the amounts are expressed in grams of each amino acid per kilogram of hydrolysate.6.- A casein hydrolysate for use according to any one of claims 1 to 5, wherein it is for use in the prevention of colorectal cancer in patients with Crohn’s disease.7.- A casein hydrolysate for use according to any one of claims 1 to 6, wherein it is for use in combination with at least one drug for use in the treatment and / or prevention of Crohn’s disease.8.- A casein hydrolysate for use according to any one of claims 1 to 7, wherein the casein hydrolysate is for oral administration.9.- A casein hydrolysate for use according to claim 8, wherein the casein hydrolysate is for administration according to a daily dose comprised between 3 and 40 g, preferably comprised between 5 and 30 g.10.- A casein hydrolysate for use according to claim 9, wherein a unit oral dose of between 4 and 8 g of the casein hydrolysate is used, which is administered from 1 to 5 times daily.11.- A casein hydrolysate for use according to any one of claims 1 to 10, wherein the casein hydrolysate is in the form of a pharmaceutical composition comprising a therapeutically effective dose of the hydrolysate and at least one pharmaceutically acceptable excipient and / or carrier.12.- A casein hydrolysate for use according to claim 11 , wherein the pharmaceutical composition is a composition suitable for oral administration.13.- A casein hydrolysate according to claim 12, wherein the pharmaceutical composition is in powder or granulate form.14.- A casein hydrolysate for use according to claim 13, wherein the composition is dosed in monodose sachets containing between 4 and 8 g of the casein hydrolysate.

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