MX2008004479A - A dairy derived composition enriched in tgf-î²for treating inflammation - Google Patents

Abstract

The present application relates to a dairy-derived composition and methods therefore or uses thereof in the prophylaxis and treatment of inflammatory diseases, including chronic intestinal inflammation, inflammation and related disorders. The composition comprises TGF-β1, TGF-β2, IGF-1 and beta-lactoglobulin and has a protein concentration of at least 15%(w/w) whey derived proteins.

Description

COMPOSITIONS TO TREAT INFLAMMATION TECHNICAL FIELD The present application is related to the treatment of inflammation and especially chronic intestinal inflammation. The present application relates more particularly to a non-toxic and topical oral formulation that includes a pharmaceutically effective amount of milk proteins and its use in the treatment of inflammation. BACKGROUND OF THE INVENTION Sumerian and ancient Egyptian cultures described inflammatory processes as abscesses and ulcers. Greek culture introduced the term edema and the vocabulary of inflammation began its development. The first full description of the inflammatory symptoms can be found in the writing of Aulus Celsus (died in 38 AD). He introduced the cardinal symptoms of inflammation: redness, swelling, heat and pain. Inflammation is usually a local protective reaction that serves to protect the body against infections or trauma. Excessive inflammation or unusual inactivity of an otherwise normal inflammatory process leads to inflammatory diseases. Inflammatory processes are a combination of very complex mechms in which many factors are involved. These factors are cellular (endothelial cells, macrophages, leukocytes, etc.) and / or humoral (coagulant factors, cytokines, eicosanoid acids, adhesion molecules, free radicals, etc.). In general, inflammatory processes are associated with immune cell proliferation, cell infiltration and accumulation of exudates at the site of inflammation and degradation of injured tissue. The release of mediators such as cytokines and eicosanoid acids leads to a more global reaction such as fever. At the end of the inflammation process, the injured tissue is destroyed by the orgm. Exposure of mammalian tissue or cells to harmful stimuli such as lipopolysaccharides (LPS, a compound found in the bacterial structure), toxic chemicals, radiations or inflammatory mediators, are known to activate immune cells to produce inflammatory mediators such as α-factor a tumor necrosis (TNF-a), interleukin-1 (IL-1), IL-β, prostaglandins (PG), leukotrienes and nitric oxide (NO). Activated immune cells can eventually lead to inflammatory diseases (eg, arthritis and septic shock), diseases linked to immunity (eg, graft rejection, autoimmune diseases and diabetes mellitus) and the death of some cell types (eg, neurons).

Prostaglandins are molecules that originate from arachidonic acid and are involved in various activities and physiological pathologies. Its production is regulated by the enzyme cyclooxygenase (COX). Non-steroidal anti-inflammatory drugs (NSAIDs) and analgesic agents such as aspirin and indomethacin, which inhibit the synthesis of prostaglandins by inhibiting COX activity, are known to provide an excellent therapeutic effect for diseases related to inflammation. Activation of the nuclear factor kB (NF-kB) is required for the production of NO, prostaglandins and TNF-a. NF-kB are also known to regulate the expression of a variety of proteins involved in various cellular responses such as apoptosis, immune responses and inflammatory reactions. In the abnormal processes of inflammation, inflammatory processes remain out of control and induce persistent diseases. Many diseases are described with an important inflammatory component, but chronic inflammatory diseases are recognized as being polysystemic diseases with an unknown origin. They are as diverse as psoriasis, Crohn's disease, rheumatoid arthritis. In general, an immune component can be observed in all these diseases.

The description of Crohn's disease done hereafter should be observed only as an illustration of the topic and will not represent a limit to the possible applications of this application. Crohn's disease is an inflammatory bowel disease that causes the persistent or recurrent inflammation of one or more parts of the intestine. Crohn's disease can affect any part of the gastrointestinal system, from the mouth to the anus. There is no known cure for Crohn's disease. The causes are unknown and not only due to stress as once thought. Inflammatory bowel diseases (IBD) can occur at any age, but usually begin in young or young adults. It is sometimes associated with medical problems outside the intestine, including arthritis, cancer, gallstones, kidney stones and skin diseases. Symptoms of Crohn's disease include abdominal pain, diarrhea and weight loss, the most frequent pain felt around the navel or the lower right part of the abdomen. Pains can occur during a meal or immediately after a meal. Constant dull pains of the abdomen may also be felt and may worsen with activities such as running and sports. Other symptoms of Crohn's disease that patients have noticed are fatigue and loss of appetite. Drugs that have mesalamine as a main ingredient are often first prescribed to a patient with Crohn's disease. Mesalamine helps control inflammation. However, possible side effects of mesalamine preparations include nausea, vomiting, heartburn, diarrhea and headache. Corticosteroids can also be given to these patients. Patients who take corticosteroids to control inflammation face a difficult dilemma between effective treatment and serious side effects. For example, corticosteroids increase the likelihood of getting infections. Crohn's disease can also be controlled with drugs that suppress the immune system such as β-mercaptopurine and azathioprine. Immunosuppressive agents block the immune reaction related to inflammation. These drugs can have side effects such as nausea, vomiting and diarrhea. In addition, drugs can decrease the patient's resistance to infection. The combination of immunosuppressive drugs and corticosteroids allows the dose of corticosteroids to be reduced over time. It is evident that immunosuppressive drugs can increase the effectiveness of corticosteroids. Infliximad is a drug recently approved to treat patients with Crohn's disease. The purpose of infliximad is to prevent inflammation by inhibiting the activity of TNF. An important medical necessity is the effective treatment of inflammation and autoimmune diseases. These diseases are treated with drugs that have high side effect profiles and limited efficacy. Currently, the commonly used therapeutic alternatives available to treat chronic inflammations consist of the use of corticosteroids or NSAIDs. Unfortunately, all of these anti-inflammatory agents are associated with highly significant side effects, including gastrointestinal irritations and bleeding, osteopenia and fluid retention, some of which can significantly reduce the well-being of patients. Several drugs have been designed to treat inflammatory diseases by targeting specific cells, specific cytokines, or specific interactions between ligands and receptors. The main advantage of these biological agents over cyclosporine and methotrexate is the absence of nephrotoxicity and hepatoxicity, but the toxic effects can take years to develop. Current treatments are directed toward either cell proliferation or the immune component of inflammation, for example, in procedures for the treatment of psoriasis. Japanese patent application JP 6145069 describes angiogenesis inhibitors comprising ganglioside GM3 or a GM3 analog as an active agent. In 100 μg / ml of GM3 the growth of normal human endothelial cells was inhibited (4.5xl04 on day 5, compared to 76x104 in the controls) US Patent No. 5, 330,977 describes n-deacetyl-lysoganglioside derivatives for use as phospholipase 2 inhibitors for the treatment of proliferative and autoimmune diseases, including various forms of cancer, psoriasis and rheumatoid arthritis. Anti-i-inflammatory agents such as glucocorticoids or aspirin inhibit the activation of NF-kB and / or repress the production of NO or prostaglandin. Some herbal drugs widely used as anti-inflammatory agents in oriental natural medicine are known to exert their effects by inhibiting the activity of NF-kB (Kopp and Ghosh, Science, 265: 956-959 (1994)).; Ray and Prefontaine, Proc. Na ti. Acad. Sci. U. S A. 91, 752-756 (1994)). There are several drugs that show inhibitory activity for the production of TNF-a, including IL-4, IL-10 and that transform the growth factor-β (TGF-β) and clinical trials that use the antibody against TNF are on the way -a and the soluble TNF-a receptor. As synthetic compounds, for example, new derivatives of 1-thiadibenzoazulene (patent of US Pat. No. 6,897,211) and dibenzoazulene compounds (patent of US Pat. No. 6,897,211) have been developed for their inhibitory activity of TNF-a. Because these agents are TNF-a inhibitors, concerns have been raised about the risk of infection resulting from such immunosuppression. Inhibitors of NO production have focused on the development of agents that specifically inhibit the enzymatic activities of inducible NO synthase (NOSi), the enzyme responsible for the synthesis of NO. For example, analogues of NO precursors such as L-arginine and L-citrulline and the aminoguanidine and isothiourea derivatives are under investigation (Babu and Griffith, Curren t Opinion in Chemical Biology, 2, 491-500 (1988) ). The derivatives of curcumin, capsaicin and caffeic acid and some other natural compounds have been reported to inhibit the activation of NF-kB, while dexamethasone, a representative of the anti-inflammatory agent of the glucocorticoid family, inhibits the production of NO by inhibiting the expression of the NOSi gene. The molecular mechanism of dexamethasone in the synthesis of NO has been known to be the suppression of transcription of the NOSi gene, concerning the inhibition of NF-kB activation (De Vera et al., Am. J. Physiol., 273, 1290-1296 (1977)).

Approximately 200 diterpene compounds have been isolated from Isodon japoni cus or near species. However, the indication that these compounds not only inhibit the production of NO, PGE2 or TNF-a in cell lines stimulated by LPS but also that these compounds can inhibit the activation of NF-kB has not been reported. The U.S. Patent 6,894,073 described camebanin, camebacetal A, camebacaurin and exisanin A, which are members of the family of caurane diterpene compounds isolated from Isodon japonicus, to inhibit not only the activation of NF-kB, but also the production of NO , prostaglandin and TNF-a in macrophage cells stimulated by LPS. These four compounds have been proposed as therapeutic agents for the treatment of inflammatory diseases, immunological diseases and cancers. Human and bovine milks contain many biologically active polypeptides that include growth factors (West, Exp. Clin. Endocrinol 8: 145-146, (1989)). One of these factors, the milk growth factor (FGM), was recently identified as being identical or having homology close to members of the TGF-β family, notably TGF-β2 (Cox et al., £ ur J. Biochem. : 353-358, (1991)). TGF-ßs are a group of polypeptides consisting of at least 5 distinct but closely related members that have considerable structural and biological homologies (Roberts, AB et al., In: Peptide Growth Factors and their Receptors, Vol. 1, pp. 419 -472, Eds. Sporn MB et al., Springer (1990)). TGF-ßs are homodimeric proteins of approximately 25 kDa comprising chains of identical 12.5 kDa polypeptides linked through disulfide bridges. They can form latent complexes with other proteins and these complexes can be activated by acid treatment or moderate proteolysis (Roberts, A. B. et al., Supra). They are multipotent, having a number of biological activities dependent on the type of the target cell, the state of differentiation of the cell and the presence of other factors. Its effects include stimulation or inhibition of cell proliferation and differentiation, regulation of extracellular matrix deposition, immunomodulation, steroidogenesis and angiogenesis (Roberts, A. B. et al., Supra). Suitable methods for purifying TGF-ßs from various species such as human, mouse, green monkey, porcine, bovine and chicken and various cellular sources such as bone cells, platelets or placental cells, as well as for produced in recombinant cell cultures and to determine their activities (European Patents 200,341; 169,016; 268,561 and 267,463; Patent Application GB 2,146,335: US Patent 4,774,322). TGF-β is multifunctional, since it can either stimulate or inhibit proliferation, cell differentiation, and other critical processes in cellular functions (M. Spor, Science, 233: 532-534 (1986)). TGF-β has been shown to have numerous regulatory actions on a wide variety of cells. Recent studies indicate an important role for TGF-β in immune cells (J. Kehrl et al., J. Exp. Med., 163: 1037-1050 (1986); HJ. Ristow, Proc. Na ti. Acad. Sci. U. S. A., 83: 5531-5533 (1986), A. Rook et al., J. Immunol., 136: 3916-3920 (1986)) and in the proliferation of soft connective tissues for healing applications. of Wounds (U.S. Patent No. 4,774,228). In addition, TGF-β has been described as a suppressor of cytokine production (interferon- (IFN-α), TGF-β), indicating a possible use as an immunosuppressant to treat inflammatory disorders (European Patent 269,408; 4,806,523). In addition, TGF-β induces the secretion of collagen in human fibroblast cultures (Roberts et al., Proc. Nati, Acad. Sci. U.S.A. 83: 4167-4117 (1986)), stimulates the release of PGs. and the mobilization of calcium (A. Tashjian et al., Proc. Na ti. Acad. Sci. U. S. A., 82: 4535-4538 (1985)) and inhibits endothelial regeneration (R. Heimark et al., Science, 233: 1078-1080 (1986)). TGF-β is produced by both immune and non-immune cells and exhibits a wide range of functions, including the modulation of immune responses. The oral administration of TGF-β2 is well known to decrease inflammatory processes. For example, Fell et al., (Aliment Pharmacol Ther 14 (3): 281-289, (2000)) have demonstrated the anti-inflammatory effect of a diet rich in polymeric TGF-β2 (CT3211: Nestlé Vevey, Switzerland) in children with IBD. In this study, 29 children with IBD were treated with CT3211 for 8 weeks. After the treatment, the results showed that 79% of the children were in complete remission of their disease. Macroscopic and histological healing of the ilium and colon was associated with a decrease in mRNA of pro-inflammatory cytokines (IL-lß, IFN-β, IL-8) in ileum and colonic mucosa as well as a mRNA of TGFβ-2 increased in the tissue of the ileum. This study confirms the interest in using the TGF-β2 solution as a therapeutic agent. TGF-ßs, including TGF-ß2 have a number of immunoregulatory properties and act in various stages of inflammatory and immune reactions. For example, they inhibit the proliferation of T and B lymphocytes (Kerhl, J.H. et al., J. Immunol., 137: 3855-3860, (1986)).; Kerhl, J.H. et al., J. Exp. Med. 163: 1037-1050 (1986)). They also antagonize the effects of interieucins including IL-1, IL-2 and IL-3 and other immunoregulatory agents such as TNF-α and interferons (Roberts, A. B. et al., Supra). TGF-ßs interfere with certain important secondary cellular functions in the presentation of the antigen and specifically demonstrated to suppress the expression of the highest histocompatibility complex (MHC) -Class II by melanomas, neuroglyclics and astrocytes (Czarniecki, et al., J. Immunol. 140: 4217-4223, (1988), Schlusener, J. Neuroimmunol, 24: 41-47, (1990), Zuber, et al., Eur. J. Immunol, 18: 1623-1626, (1988)). Regulation of MHC-Class II expression on epithelial cells in the intestine by TGF-ßs or MGF has been reported in European patent EP0527283. The patent of E.U. 6,649,168 discloses a dry powder pharmaceutical composition comprising a TGF-β in a water soluble salt (calcium chloride, calcium phosphate, potassium acetate, lithium acetate, ammonium acetate or ammonium bicarbonate) for therapeutic applications such as the promotion and acceleration of wound healing, bone repair, soft tissue repair, nerve repair, stroke therapy, cancer treatment, bone marrow protection, cardioprotection, induction of angiogenesis, regulation of mammalian cell growth and induction of anti-inflammatory and immunosuppressive activity. In the patent of E.U. 6,403,562 methods for treating diseases related to autoimmune diseases, such as multiple sclerosis (MS) are presented. These methods comprise administering IL-10 and TGF-β in combination to a person affected or predisposed to an autoimmune disease. When administered in combination, these cytokines act in a synergistic manner as suppressive factors in order to inhibit the activation of self-reactive T cells that are involved in autoimmune diseases. The U.S. Patent No. 5,461,033 discloses the use of an isolated casein acid fraction isolated from bovine milk and containing TGF-β2 in the treatment of Crohn's disease. The data reported in the patent is based on in vitro experimentation and the patent does not give the specific description of the nutritional formulas that can be used in the treatment of Crohn's disease. The U.S. Patent 5,952,295 gives an accurate formulation of the enteric preparation for the treatment of inflammatory conditions in the gastro-intestinal tract using a casein fraction with TGF-β2 and reports the improvement of such inflammatory conditions in an open study with 11 young patients. European Patent 0313515 describes a fraction of the milk growth factor obtained from milk and the use of this fraction to treat trauma, suppress the immune response or treat cancer in mammals. The patent application WO 02705828 proposes the treatment of dermatological disorders by means of neutral or basic growth factors originating from milk or colostrum. Japanese patent 14308090 describes a colostrum fraction with TGF-β activity.

A milk fraction of the growth factors obtained from the chromatographic methods, especially with an ion exchange step is described in the U.S. Patent. 6,447,808. This fraction can be used for the treatment of gastrointestinal lesions, ulcers or superficial wounds. Patent application WO 01/25276 also describes the isolation of growth factors from dairy products by the use of chromatography. Different fractions can be produced by this process, especially fractions enriched with TGF-β and IGF-β. Patent application WO 03/006500 describes a process for obtaining a milk fraction enriched with TGF-β. This process is based on a stage of precipitation induced by a thermal treatment in acidic conditions followed by the recovery of the precipitate by filtration. The fraction contains 45 to 80% of alpha-lactalbumin protein and more than 5 μg of TGF-β per gram of protein. This fraction is proposed for the treatment of autoimmune diseases, arthritis, osteoporosis, wound healing and graft rejection. Considering the state of the art described above, all the methods of the prior art are complex, expensive and slow. Thus, there is still a significant need for a new and simple method to obtain a milk fraction enriched with TGF-β that can be used effectively in the treatment of chronic inflammatory diseases (such as chronic intestinal diseases) without the disadvantages associated with the treatments currently available. Products containing growth factors, especially TGF-β2, present a very interesting procedure for the treatment of chronic inflammatory disease and especially Crohn's disease. SUMMARY OF THE INVENTION One purpose of the present application is to provide a new and simple method for obtaining a milk fraction enriched with TGF-β that can be used as an effective treatment for chronic inflammatory bowel disorders, a composition containing such fraction and uses of the same According to the present application there is provided a dairy-derived composition for the prophylaxis or treatment of an inflammatory disease or a disorder related to a subject, wherein said composition comprises TGF-β1, TGF-β2, IGF-1, beta- lactoglobulin and at least 15% (w / w) of proteins derived from the whey. Preferably the TGF-β1 concentration is between about 0.1 and 5 μg per gram of the composition and more preferably between about 0.2 and 1.2 μg per gram of the composition. In one embodiment of the application, the concentration of TGF-β2 is preferably between about 5 and 50 μig per gram of the composition and more preferably between about 10 and 18 μig per gram of the composition. In one embodiment, the concentration of the proteins derived from the whey is at least 50% (w / w) and more preferably at least 80% (w / w). The composition may further comprise between 0.1 and 15% of minerals. In one embodiment, the composition comprises at least 35% and preferably at least 70% water soluble proteins. The inflammatory disease may be for example a chronic inflammatory disease or chronic inflammatory bowel disease, such as Crohn's disease or ulcerative colitis. The subject can be an animal or a human. According to the present application, there is provided the use of a dairy-derived composition comprising TGF-β1, TGF-β2, IGF-1, beta-lactoglobulin and at least 15% (w / w) of the proteins derived from the serum of milk for the prophylaxis or treatment of inflammatory diseases or related disorders. Still according to the present application, there is provided a method for treating or preventing an inflammatory disease or a related disorder, comprising the step of administering to a subject a therapeutically effective amount of a dairy-derived composition comprising TGF-βl., TGF-ß2, IGF-1, beta-lactoglobulin and at least 15% (w / w) of proteins derived from whey. Still according to the present application, there is provided a method for treating or preventing an inflammatory disease or related disorder, comprising the step of • combining TGF-β1, TGF-β2, IGF-1, beta-lactoglobulin and at least 15% (p / p) of the proteins derived from the whey to obtain a composition derived from dairy products; Y • administer a therapeutically effective amount of said dairy-derived composition to a subject in need thereof. One skilled in the art will recognize that various types of chronic inflammatory diseases can be avoided or treated with the method and composition of the present application. For example, without limitation, the term chronic inflammatory disease includes rheumatoid arthritis, psoriasis, skin inflammation, atopic dermatitis, encephalitis, allergic alveolitis, chronic pulmonary inflammation, ischemia-revascularization injury, systemic lupus erythematosus, myositis, ankylosing spondyloarthritis, scleroderma, polyradiculoneuropathy acute inflammatory disease, vasculitis, appendicitis, arachnoiditis, myocarditis, acute cholecystitis, chronic obstruction of the respiratory tract, chronic hepatitis, chronic obstructive pulmonary disease, conjunctivitis, dermatitis, enteritis, gingivitis, hepatitis, ileitis, asthma and / or intestinal inflammatory diseases Chronic diseases such as Crohn's disease, inflammatory bowel disease or ulcerative colitis. Accordingly, one aspect of the present application contemplates a method for reducing the effects of a proliferative and / or inflammatory disease in a subject, the method comprising treating a chronic inflammatory disease with an effective amount of a composition capable of preventing or treating inflammatory diseases. Chronicles and related disorders, as described herein. For the purpose of the present application, the following terms are defined below. The term "inflammation" as used herein is proposed to represent the normal response of the immune system to infection or irritation. The present application is directed only to the cellular compartment of the immune response, also referred to as the Thl component of the immune system and does not target the humoral or exudative portion of the immune response, also referred to as the Th2 component of the immune system. The terms "inflammatory disease" and "inflammatory disorders" as used herein are proposed to refer to any disease or disorder that affects the normal response of the immune system. The disease or disorder may involve either an excessive or insufficient response of the immune system to infection or irritation, a lack of response after infection or irritation, and a response in the absence of infection or irritation. Such disorders may be, for example, without limitation, ulcerative proctitis, proctosigmoiditis, left lateral colitis, pan-ulcerative colitis (total), psoriatic arthritis, ankylosing spondyloarthritis, juvenile ankylosing spondyloarthritis, seronegative enthesopathy, arthropathy syndrome, Reiter syndrome, spondyloarthropathies and / or endogenous uveitis. The term "inflamed tissue" as used herein refers to a tissue affected by inflammation and affected cells contained within the tissue. Cells that have migrated to the site of inflammation, such as leukocytes, are excluded from the definition of this term. Preferably, the inflamed tissue is of a mammalian species, preferably a human.

The terms "effective amount" and "therapeutically effective amount" as used herein are proposed to refer to an amount sufficient to initiate a beneficial or desired clinical result, such as an improvement of the patient's condition. An effective amount may be administered in one or more doses. For the purposes of this application, an effective amount of the present composition is an amount that induces a therapeutic or prophylactic response against at least one factor responsible for inflammation. Such amount will vary according to the nature of the inflamed tissue, the severity of the inflammation, the mode of administration, etc. A person skilled in the art can easily and easily monitor the inflamed tissue in order to determine what will be such an effective amount. The terms "individual", "subject" or "patient" according to this application are proposed to refer to a vertebrate, preferably a mammal, more preferably a human. Mammals include, but are not limited to, farm animals, sports animals, rodents, primates and pets. The term "minerals" as used herein is proposed to refer to constituents and saline minerals normally found in milk. For example, calcium, phosphorus, magnesium, potassium, sodium, chlorine, sulfur and citric acid can be used in various concentrations. This also means that the microelements known in the matter present in milk can be assimilated or inferred therein. The term "treatment" as used herein is proposed to mean a medical remedy to improve or alleviate in part or totally the condition of a subject suffering from inflammation or inflammatory disorders. The term "prophylaxis" as used herein is proposed to refer to any procedure directed at the prevention of inflammation or inflammatory disorders. It will be understood that this term represents both primary prophylaxis, i.e., the prevention of the onset of the disease or disorder and secondary prophylaxis, i.e., prevention of the aggravation of a disease or disorder already developed. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the dose-dependent inhibition of the proliferation of murine ConA splenocytes induced by the composition of the present application; Figure 2 shows the dose-dependent inhibition of the production of murine splenocytes IL-2, ConA induced by the composition produced in example 1 and described in example 2; Figure 3 shows the dose-dependent inhibition of the production of murine splenocytes IFN-α. ConA induced by the composition produced in Example 1 and described in Example 2; Figure 4 illustrates the effect of colorectal administrations of the composition produced in Example 1 and described in Example 2 on daily body weight variations of Sprague-Dawley rats treated with 2,4,6-trinitrobenzenesulfonic acid (TNBS), an inducer inflammatory chemical, as described in detail in example 4; Figure 5 illustrates the effects of colorectal administrations of the composition produced in Example 1 and described in Example 2 on the weight-to-length ratios of the colons of the Sprague-Dawley rats treated with TNBS, as described in FIG. detail in example 4; Figure 6 illustrates the effects of the colorectal administrations of the composition produced in example 1 and described in example 2 on the macroscopic damage of rat colon tissues previously irritated with TNBS, as described in detail in example 4. Figure 7 illustrates the effects of the composition produced in example 1 and described in example 2 on the macroscopic appearance of lesions in TNBS colitis, as described in detail in example 4.

Figure 8 shows the levels of hematocrits before and during oral treatment with different doses of the composition produced in example 1 and described in example 2, wherein A is a placebo, B is the composition (7.82 mg / kg), C is the composition (78.2 mg / kg) and D is the composition (782 mg / kg), as described in detail in example 5; and Figure 9 shows the hemoglobin levels before and during oral treatment with different doses of the composition produced in example 1 and described in example 2, wherein A is a placebo, B is the composition (7.82 mg / kg) , C is the composition (78.2 mg / kg) and D is the composition (782 mg / kg). DETAILED DESCRIPTION OF THE PREFERRED MODALITIES The present application will now be described in more detail hereafter with reference to the accompanying drawings, in which the preferred embodiments of the application are shown. This request can, however, be incorporated in many different forms and should not be construed as limited to the modalities set forth herein; rather, these modalities are provided, so that this description is thorough and complete and completely transmits the scope of the request to experts in the field. All patents, patent applications, articles and publications mentioned herein, both supra and infra, are incorporated herein by reference. According to the present application, there is provided a composition and method for treating chronic inflammatory diseases in an individual, wherein the composition is effective to stimulate a specific immune response against an aberrantly expressed physiological imbalance in inflamed tissues. This composition comprises a variety of products that share the characteristics of the growth and regeneration of a growth factor that is aberrantly expressed in inflamed tissues (such as inflamed human tissues). The particular growth factors included in the composition of the present application are, but are not limited to, TGFs, such as TGF-β1 and TGF-β2, beta-lactoglobulin and at least 15% of proteins derived from dairy products. The growth factors present in the composition are distinguished by having a strong anti-inflammatory action. The concentration of the compounds in the composition of the present application can be adjusted depending on the needs. TGF-β1 can be present at a concentration of between 0.1 to 5 μg / g of the composition and TGF-β2 can be present at a concentration of between 5 and 50 μg / g of the composition. Preferably, TGF-β1 is present at a concentration of 0.2 to 1.2 μg / g and TGF-β2 between 10 to 18 μg / g of the composition. The preparation can be used daily in the usual way. In another embodiment of the present application, the composition comprises at least 70% (w / w) of the proteins derived from the whey. The total concentration of the proteins in the composition would normally be at least 80%. The concentration of fat can generally be at a concentration between 0.5 to 10% (w / w) in the composition. In a particular embodiment of the present application, at least 35%, preferably 70% of the proteins are soluble in water. The proteins may be comprised of at least 60% (w / w) of β-lactoglobulin and / or between 0.1 to 30% (w / w) of lactoferrin. The active compounds according to the application, such as the aforementioned growth factors, can be mixed with pharmaceutically tolerable diluents or vehicles customary and, if appropriate, with other auxiliary molecules and administered, for example, topically, orally, parenterally or colorectally. This formulation can preferably be administered orally in the form of granules, capsules, pills, tablets, film-coated tablets, sugar-coated tablets, syrups, emulsions, suspensions, dispersions, aerosols, solutions and / or liquids. They can also be administered as suppositories, vaginal suppositories and / or parenterally e.g., in the form of solutions, emulsions, creams or suspensions. It can be administered in preparation for delayed time release or protected against gastric acid by coating to be released in the intestinal part of the digestive tract. The preparations to be orally administered may contain one or more additives such as sweeteners, flavoring agents, colorants and preservatives. The tablets may contain the composition mixed with common pharmaceutically tolerated auxiliaries, such as, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose and talc.; granulation agents and agents that promote the disintegration of tablets in oral administration, such as starch or alginic acid; binding agents such as starch or gelatin; and lubricants such as magnesium stearate, stearic acid and talc. The formulations are prepared for example by diluting the composition with solvents and / or excipients if appropriate, using emulsifiers and / or dispersants, it being possible for example, in the case of the use of water as a diluent to optionally use organic solvents as auxiliary solvents.

The administration is carried out in the customary manner preferably orally or parenterally or prelingually, sublingually, colorectally, topically or intravenously. In the case of oral administration, apart from the mentioned excipients, the tablets may also contain additives, such as sodium citrate, calcium carbonate and calcium phosphate together with other various additives, such as starch, preferably potato starch, gelatin and the similar. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate and talc can be used additionally to make tablets. In the case of aqueous suspensions and / or elixirs, which are proposed for oral administration, the active compounds can be mixed, in addition to the aforementioned auxiliaries, with various flavor improvers or colorants. In the case of parenteral administration, the solutions of the composition can be employed using suitable liquid excipients. The capsules may contain the composition as a single constituent or be mixed with a solid diluent such as calcium carbonate, calcium phosphate or kaolin. The injectable formulations are also formulated in a manner known per se. In addition, antioxidants can be added to the composition and to the pharmaceutical and topical formulations. Particularly preferred is the use of naturally occurring or naturally identical compounds such as, for example, tocopherols.

The antioxidants mentioned are contained in the compositions according to the request, for example, in amounts from 0.01-5% by weight, in particular from 0.5-2% by weight, based on the total composition. The preferred formulation according to the application can be formulated as liquid, pasty or solid preparations, for example as aqueous solutions or with alcohol, suspensions or aqueous emulsions. Estimates of the prevalence of chronic inflammatory diseases vary according to each specific disease. For example, inflammatory bowel diseases are considered to affect 0.3% of the American population. In all types of chronic inflammatory diseases, the composition can be used prophylactically and for the treatment of diseases. Although the detectable immune or tissue response is probably beneficial, efficacy can also be inferred by an improvement in symptoms or control of the chronic inflammatory condition beyond what would be expected without treatment. For prophylaxis, the composition is administered in order to decrease the manifestations of the disease in frequency and intensity. Treatment in the manifest stage leads to its reduction and relief of symptoms. The composition can also be administered colorectally or enteric-coated to be delivered to the intestinal part of the digestive tract. The composition can also be incorporated into the oral and pharmaceutical bases customary for preferred oral administrations. The application also relates to the use of the composition according to the application for the production of a pharmaceutical composition, in particular oral and pharmaceutical compositions for the prophylaxis and treatment of chronic inflammatory diseases. Without wishing to be bound by theory, the composition also addresses the underlying T cell disorder that results in an inflammatory condition. The Applicants have recognized that most if not all current therapies in the prior art for inflammatory conditions mediated by T cells are designed to eliminate T cells and thereby improve the condition of inflammation. A major problem with these treatments is that there is a possibility that the therapy itself is so toxic that it can promote recurrence during the recovery of health. The synergy of the two TGF-β factors of the composition results in a treatment non-toxic highly effective for chronic inflammatory diseases found without the side effects observed with virtually all other therapies for these diseases. The present application will be more easily understood by referring to the following examples that are given to illustrate the application instead of limiting its scope. EXAMPLE 1: Preparation of the composition They were rehydrated up to 5% in deionized water and they were stabilized overnight at 4 ° C under agitation, 1000 kg of a whey protein isolate. The pH of the solution was adjusted between 4.8 and 5.2 by the addition of concentrated HCl. This adjustment induced the precipitation of the protein. The precipitate was then recovered by centrifugation in a Westfalia separator at 25 m3 per hour and washed with acidified water. The precipitate was rehydrated to obtain 800 liters, which were thermally treated at 75 ° C for 15 seconds, before concentration and dehydrated by spraying. Approximately 60 kg of the composition was obtained. EXAMPLE 2: Analysis of the composition The composition of the ingredient is presented in Table 1. The proteins were analyzed by the Dumas method, while the fat was analyzed by the Mojonnier method, the moisture by drying in a drying oven, the beta-lactoglobulin by RP-HPLC and growth factors by ELISA methods. Table 1: Typical analysis of the composition Compound Amount Proteins 86% water soluble proteins Fat 1.9% Beta-lactoglobulin 66% Humidity 4.2% IGF-1 54 pg / mg TGF-ß2 11.700 pg / mg TGF-ßl 300 pg / mg EXAMPLE 3: Inhibition of mouse splenocyte proliferation TGF-β has multiple immunosuppressive effects at the cellular level and has been described as an inhibitor of type 1 and type 2 cells, B cells, CD8 + T cells, macrophages and natural cytolytic lymphocytes. TGF-β blocks the progression of the cell cycle and may have a direct effect on the expression of the gene encoding IL-2 (Brabletz et al., Mol.Cel. Biol., 13 (2): 1155-1162, 1993). Importantly, it can suppress the expression of IL-12 and IL-2 receptors, as well as sub-regulate MHC class II expression on macrophages (especially by antagonizing TNF-α and IFN-α) (Harber et al. ., Exp. Rev. Mol.Med., Nov. 27 http: / www-ermm.cbcu.cam.ac.uk/00002143h.htm, 2000). The importance of TGF-β in the induction and maintenance of normal tolerance has been illustrated by knockout mice with TGF-β who incur severe autoimmune, spontaneous multiple organ diseases, associated with auto-production of antibodies, which proves to be fatal in early life (Harber et al., Exp. Rev. Mol.Med., Nov. 27., http: / www-ermm. cbcu .cam.ac.uk / 00002143h. htm, 2000). T-lymphocytes isolated from murine spleen (splenocytes) are immune cells that respond to ConA (mitogenic stress) by proliferation and cytokine production (Mercier et al., In terna T. Dairy J., 14: 175-183, (2004), Wong et al., J. Dairy Sci., 81: 1825-1832 (1998)). The purpose of this example was to evaluate the effect of inhibition of the composition produced in Example 1 and described in Example 2 on the proliferation of splenocytes and the production of pro-inflammatory cytokines. BALB / c mice 6-8 weeks old were obtained from Charles River (Montreal, Canada). Mice were sacrificed by CO 2 inhalation and single cell suspensions were prepared individually from the murine spleen under aseptic conditions. The murine lymphocytes (1.25 x 106 cells / ml) were treated by ConA (1.25 μg / ml) for 72 hours. Almar blue® was added 24 hours before the measurement of the proliferation of lymphocytes induced by Con A. The levels of cytokines (IL-2 and IFN-α) in the supernatant were also determined after the incubation period. BSA was used as a protein control (1-1000 μg / ml). The composition produced in Example 1 and described in Example 2 (1, 10, 50, 100, 500 and 1000 μg of the total composition per ml of the culture medium) was added to the cells for the entire incubation period and decreased depending on dose 1) lymphocyte proliferation (Figure 1), 2) IL-2 production (Figure 2), and 3) IFN-α production (Figure 3). As shown in Figure 1, there was significant inhibition (SI) of the proliferation induced by the composition. Without wishing to be bound by the theory, these results suggest that the composition has immunomodulatory functions and that it can be used to target chronic immune-mediated inflammatory diseases such as psoriasis, inflammatory bowel diseases and related arthropathies. EXAMPLE 4: Testing on TNBS-induced inflammation in healthy rats Bowel diseases (IBD) is a term used to describe a collection of diseases involving the intestine and characterized by chronic inflammation, sometimes accompanied by ulceration in the intestine. small or large intestine. Cytokines play an important role in the modulation of the immune system as they are rapidly synthesized and secreted from activated inflammatory cells and induce the production of adhesion molecules and other inflammatory mediators, such as oxygen reactive metabolites, nitric oxide and mediators of lipids (De Winter et al., Am J Physiol 276: G1317-1321, 1999). Cytokines induce, amplify, prolong and inhibit inflammation. Recently, several murine models of colitis have been developed that have highlighted the important role that abnormalities of the immune system can play, particularly those that affect T cells in the pathogenesis of the disease. The purpose of this example was to characterize the effect of the local application of the composition produced in Example 1 and described in Example 2 on TNBS-induced colitis in rats. The male Sprague Dawley (Charles River Laboratories, Montreal) were kept in individual cages, at 20 ° C and 55% relative humidity with cycles of 12 h of light / 12 h of darkness, in a facility that meets the guidelines of the Canadian Council on Animal Care. The rats were subjected to a fasting period of 24 hours before the induction of inflammatory tension. Inflammation of the colon was induced when using a technique from Togawa et al. (Am. J. Physiol., 283: 187-195, 2002). Briefly, the rats were lightly anesthetized with isoflurane after food deprivation overnight and an 8 cm polyethylene catheter was inserted into the colon through the anus. TNBS (Sigma-Aldrich, Canada) was dissolved in 50% (vol / vol) of aqueous ethanol (25 mg / ml), injected into the colon (total volume of 1 ml / rat). The control rats received 1 ml of PBS intracolonically in place of TNBS and ethanol. The four groups of rats were treated in the following way: Group 1: Control without tension, received 2 ml of PBS (n = 2) once a day through the entire protocol; Group 2: Control with tension: TNBS (tension agent) in the intestinal lumen received 2 ml of PBS once a day, 5 days before the administration of the tension agent in the lumen and 24 hours after the administration of the agent. tension in the lumen (n = 8). Group 3: Colorectal administration of the composition produced in example 1 and described in example 2 (78.2 mg / kg in 2 ml of PBS, n = 7) once a day, 5 days before administration of the tension agent in the lumen and 24 hours after the administration of the tension agent in the lumen. The animals were weighed every day during the entire protocol time. Forty-eight hours after the administration of TNBS, the rats were sacrificed and the 1 cm extirpation of the intestine was sampled. The samples were weighed and the macroscopic colonic damage was recorded by the following scale: 0. No damage 1. Localized hyperemia, without ulcer 2. Ulcerations without hyperemia of the thickness of the bowel wall 3. Ulceration with inflammation at one site 4. Two or more sites of ulceration / inflammation 5. Major sites of damage that extend more than 1 cm along the length of the colon 6-10. If the damage extends more than 2 cm along the length of the colon, the registration is increased by one for each additional 1 cm body weight. As shown in Figure 4, the body weights of the animal were constantly increased before fasting and the injection of TNBS in all groups. After the injection of PBS (only for control animals), the animals again gained the lost body weight during the fasting period. However, in animals treated with TNBS the animals did not gain back the lost body weight during the fasting period. As the animals were treated with the composition produced in Example 1 and described in Example 2, the body weight was increased after the injection of TNBS. Weight-to-length ratio. The injection of TNBS increased the weight-to-length ratio in the colon for both the TNBS groups and that of the composition produced in example 1 and described in example 2, indicating the presence of inflammation and edema. However, the increase was significantly lower (P <0.05) in animals treated with the composition produced in Example 1 and described in Example 2 than in the animals treated with TNBS (Figure 5). Macroscopic records. The injection of TNBS increased the macroscopic records of inflammation in both groups treated with either TNBS and the composition produced in example 1 and described in example 2, indicating the presence of inflammation and edema. However, the increase was significantly lower (P <0.05) in animals treated with the composition produced in example 1 and described in example 2, than in the animals treated with TNBS (Figure 6 and Figure 7). EXAMPLE 5: HLA-B27 Rats The HLA-B27 transgenic rat, which expresses human HLA-B27 and ß2m, exhibits a phenotype similar to humans suffering from rheumatic disorders related to B27 such as reactive arthritis, ankylosing spondyloarthritis, psoriasis and inflammatory disease of the intestine. This rat model is useful for research and pharmacological studies of spontaneous systemic inflammation, arthritis, inflammatory bowel disease and skin diseases including psoriasis and alopecia. The main objective of this example was to evaluate the in vivo efficacy of the composition produced in Example 1 and described in Example 2, in IBD present in the rat model HLA-B27. Basically, four groups of ten (10) female HLA-B27 transgenic rats were formed (7 to 20 weeks of age) following randomization which was based on the week of the first symptoms of IBD (i.e., Hemoccult ™ positive). After one week of adaptation, the composition produced in example 1 and described in example 2 (7.82, 78.2 and 782 mg / kg) was orally administered daily through gastric puncture (total volume of 1.5 ml). Hema tocri tos. As shown in Figure 8, hematocrit levels were higher in the rats treated with 7.82 and 78.2 mg / kg of the composition produced in Example 1 and described in Example 2 compared to the value observed before starting the treatment. Also, animals treated with 7.82 and 78.2 mg / kg of the composition produced in example 1 and described in example 2 had a higher level of hematocrit compared to placebo (PBS). Hemoglobin: During the course of the disease, hemoglobin levels decreased significantly in all groups. However, the decrease was significantly lower in rats treated with 7.82 and 78.2 mg / kg of the composition produced in example 1 and described in example 2 compared to placebo or even with 782 mg / kg of the composition produced in the example 1 and described in example 2 (Figure 9). Although the request has been described in relation to specific modalities thereof, it will be understood that it is capable of further modifications and this request is proposed to cover any of the variations, uses or adaptations of the application following, in general, the principles of the application and including such deviations from the present disclosure as are within the practice known or accustomed in the art to which the application pertains and as it may be applied to the essential features set forth herein and as follows in the scope of the claims annexes.

Claims (58)

1. CLAIMS 1. A composition derived from dairy products for the prophylaxis or treatment of an inflammatory disease or a related disorder in a subject, wherein said composition comprises TGF-β1, TGF-β2, IGF-1, beta-lactoglobulin and at least 15% (w / w) of proteins derived from whey. The composition of claim 1, wherein the concentration of said TGF-β1 is between about 0.1 and 5 μg per gram of the composition. 3. The composition of claim 2, wherein the concentration of said TGF-β1 is between about 0.2 and 1.2 μg per gram of the composition. The composition of claim 1, wherein the concentration of said TGF-β2 is between about 5 and 50 μg per gram of the composition. The composition of claim 4, wherein the concentration of said TGF-β2 is between about 10 and 18 μg per gram of the composition. The composition of claim 1, wherein the concentration of said proteins derived from the whey is at least 50% (w / w). The composition of claim 6, wherein the concentration of said proteins derived from the whey is at least 80% (w / w). 8. The composition of claim 1, further comprising between about 0.1 and 15% (w / w) of minerals. The composition of claim 1, wherein said composition comprises at least 35% water soluble proteins. The composition of claim 9, wherein said composition comprises at least 70% water soluble proteins. The composition of claim 1, wherein the inflammatory disease is a chronic inflammatory disease. The composition of claim 11, wherein the chronic inflammatory disease is a chronic inflammatory bowel disease. The composition of claim 12, wherein the chronic inflammatory disease is Crohn's disease or ulcerative colitis. The composition of claim 1, wherein said subject is a human. 15. The use of a dairy-derived composition for the prophylaxis or treatment of an inflammatory disease or a related disorder in a subject, wherein said composition comprises TGF-β1, TGF-β2, IGF-1, beta-lactoglobulin and at least 15% (w / w) of proteins derived from whey. 16. The use of claim 15, wherein the concentration of said TGF-β1 is between about 0.1 and 5 μg per gram of the composition. The use of claim 16, wherein the concentration of said TGF-β1 is between about 0.2 and 1.2 μg per gram of the composition. The use of claim 15, wherein the concentration of said TGF-β2 is between about 5 and 50 μg per gram of the composition. The use of claim 18, wherein the concentration of said TGF-β2 is between about 10 and 18 μg per gram of the composition. The use of claim 15, wherein the concentration of said proteins derived from the whey is at least 50% (w / w). The use of claim 20, wherein the concentration of said proteins derived from the whey is at least 80% (w / w). 22. The use of claim 15, wherein said composition further comprises between about 0.1 and 15% (w / w) of minerals. 23. The use of claim 15, wherein said composition comprises at least 35% water soluble proteins. 24. The use of claim 23, wherein said composition comprises at least 70% water soluble proteins. 25. The use of claim 15, wherein the inflammatory disease is a chronic inflammatory disease. 26. The use of claim 25, wherein the chronic inflammatory disease is a chronic inflammatory bowel disease. 27. The use of claim 26, wherein the chronic inflammatory disease is Crohn's disease or ulcerative colitis. 28. The use of claim 15, where said subject is a human. 29. A method for preventing or treating an inflammatory disease or a related disorder, comprising the step of administering to a subject a therapeutically effective amount of a composition derived from dairy products comprising TGF-β1, TGF-β2, IGF-1, beta -lactoglobulin and at least 15% (w / w) of proteins derived from whey. 30. The method of claim 29, wherein the concentration of said TGF-β1 is between about 0.1 and 5 μg per gram of the composition. 31. The method of claim 30, wherein the concentration of said TGF-β1 is between about 0.2 and 1.2 μg per gram of the composition. 32. The method of claim 29, wherein the concentration of said TGF-β2 is between about 5 and 50 μg per gram of the composition. The method of claim 32, wherein the concentration of said TGF-β2 is between about 10 and 18 μg per gram of the composition. 34. The method of claim 29, wherein the concentration of said proteins derived from the whey is at least 50% (w / w). 35. The method of claim 34, wherein the concentration of said proteins derived from the whey is at least 80% (w / w). 36. The method of claim 29, wherein said composition further comprises between about 0.1 and 15% (w / w) of minerals. 37. The method of claim 29, wherein said composition comprises at least 35% water soluble proteins. 38. The method of claim 37, wherein said composition comprises at least 70% water soluble proteins. 39. The method of claim 29, wherein the inflammatory disease is a chronic inflammatory disease. 40. The method of claim 39, wherein the chronic inflammatory disease is a chronic inflammatory bowel disease. 41. The method of claim 40, wherein the chronic inflammatory disease is Crohn's disease or ulcerative colitis. 42. The method of claim 29, wherein said subject is a human. 43. The method of claim 29, wherein said administration is selected from the group consisting of oral administration, parenteral administration, topical administration and colorectal administration. 44. A method for preventing or treating an inflammatory disease or related disorder in a subject, comprising the step of • combining TGF-β1, TGF-β2, IGF-1, beta-lactoglobulin and at least 15% (w / w) ) of proteins derived from whey to obtain a composition derived from dairy products; and • administering a therapeutically effective amount of said dairy-derived composition to a subject in need thereof. 45. The method of claim 44, wherein the concentration of said TGF-β1 is between about 0.1 and 5 μg per gram of the composition. 46. The method of claim 45, wherein the concentration of said TGF-β1 is between about 0.2 and 1.2 μig per gram of the composition. 47. The method of claim 44, wherein the concentration of said TGF-β2 is between about 5 and 50 μg per gram of the composition. 48. The method of claim 47, wherein the concentration of said TGF-β2 is between about 10 and 18 μg per gram of the composition. 49. The method of claim 44, wherein the concentration of said proteins derived from the whey is at least 50% (w / w). 50. The method of claim 49, wherein the concentration of said proteins derived from the whey is at least 80% (w / w). 51. The method of claim 44, wherein said composition further comprises between about 0.1 and 15% (w / w) of minerals. 52. The method of claim 44, wherein said composition comprises at least 35% water soluble proteins. 53. The method of claim 52, wherein said composition comprises at least 70% water soluble proteins. 54. The method of claim 44, wherein the inflammatory disease is a chronic inflammatory disease. 55. The method of claim 54, wherein the chronic inflammatory disease is a chronic inflammatory bowel disease. 56. The method of claim 55, wherein the chronic inflammatory disease is Crohn's disease or ulcerative colitis. 57. The method of claim 44, wherein said subject is a human. 58. The method of claim 44, wherein said administration is selected from the group consisting of oral administration, parenteral administration, topical administration and colorectal administration.