WO2007038870A1 - Composition derivee du lait enrichie en tgf-$g(b) pour le traitement de l'inflammation - Google Patents

Composition derivee du lait enrichie en tgf-$g(b) pour le traitement de l'inflammation Download PDF

Info

Publication number
WO2007038870A1
WO2007038870A1 PCT/CA2006/001637 CA2006001637W WO2007038870A1 WO 2007038870 A1 WO2007038870 A1 WO 2007038870A1 CA 2006001637 W CA2006001637 W CA 2006001637W WO 2007038870 A1 WO2007038870 A1 WO 2007038870A1
Authority
WO
WIPO (PCT)
Prior art keywords
composition
tgf
concentration
inflammatory disease
per gram
Prior art date
Application number
PCT/CA2006/001637
Other languages
English (en)
Inventor
Eric Lamiot
Réjean DROUIN
Patrice E. Poubelle
Christina Juneau
Original Assignee
Advitech Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Advitech Inc. filed Critical Advitech Inc.
Priority to NZ567126A priority Critical patent/NZ567126A/en
Priority to US11/695,757 priority patent/US7763257B2/en
Publication of WO2007038870A1 publication Critical patent/WO2007038870A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/20—Milk; Whey; Colostrum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A61K38/1722—Plasma globulins, lactoglobulins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18—Growth factors; Growth regulators
    • A61K38/1841—Transforming growth factor [TGF]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/30—Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]

Definitions

  • the present application is related to the treatment of inflammation, and especially chronic intestinal inflammation.
  • the present application more particularly relates to a non-toxic oral and topical formulation that includes a pharmaceutically effective amount of dairy proteins and its use in the treatment of inflammation.
  • Inflammation is usually a local protective reaction that serves to protect the organism 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 mechanisms in which many factors are implicated. These factors are cellular (endothelial cells, macrophages, leucocytes, etc) and/or humoral (coagulation factors, cytokines, eicosanoid acids, adhesion molecules, free radicals, etc). In general, inflammatory processes are associated with immune cell proliferation, infiltration of cells and accumulation of exudates at the inflammation site, and degradation of the injured tissue. Liberation of mediators like cytokines and eicosanoid acids lead to a more global reaction like fever. At the end of the inflammation process, the injured tissue is destroyed by the organism.
  • LPS lipopolysaccharides
  • IL-1 interleukin-1
  • PG prostaglandins
  • NO nitric oxide
  • Activated immune cells can eventually lead to inflammatory diseases (e.g., arthritis and septic shock), diseases linked to immunity (e.g., graft rejection, autoimmune diseases, and diabetes mellitus), and the death of some cell types (neurons for example).
  • Prostaglandins are molecules originating from arachidonic acid and involved in various physiological activities and pathologies. Their production is regulated by the enzyme cyclooxygenase (COX).
  • COX cyclooxygenase
  • NF-KB nuclear factor KB
  • NF- ⁇ B nuclear factor KB
  • Crohn's disease is an inflammatory bowel disease that causes persistent or recurring 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 are not only due to stress as was once thought. Inflammatory bowel diseases (IBD) can occur at any age, but usually begins in the late teens or early adulthood. It is sometimes associated with medical problems outside of the intestine, including arthritis, cancer, gallstones, kidney stones, and skin conditions. Crohn's disease symptoms include abdominal pains, diarrhea, and weight loss. Pain is most often felt around the navel or the lower right part of the abdomen. The pains may occur during eating or soon after a meal. Steady dull aches of the abdomen may also be felt and may be worsen with activities like jogging and sports. Other Crohn's disease symptoms that patients have noted are fatigue and loss of appetite.
  • IBD Inflammatory bowel diseases
  • Drugs having mesalamine as a main ingredient are often first prescribed to a patient with Crohn's disease.
  • Mesalamine helps in controlling the inflammation.
  • possible side effects of mesalamine preparations include nausea, vomiting, heartburn, diarrhea, and headache.
  • Corticosteroids may also be administered to those patients. Patients taking corticosteroids to control inflammation face a difficult trade-off 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 suppressing the immune system such as 6-mercaptopurine and azathioprine.
  • Immunosuppressive agents block the immune reaction related to inflammation. These drugs may have side effects like nausea, vomiting, and diarrhea. In addition, the drugs may lower patient's resistance to infection.
  • immunosuppressive drugs can increase the effectiveness of corticosteroids.
  • Infliximad is a recently approved drug for treating Crohn's disease patients. The purpose of infliximad is to prevent inflammation by inhibiting the activity of TNF.
  • Japanese patent application JP 6145069 describes angiogenesis inhibitors comprising ganglioside GM3 or a GM3 analog as an active agent. At 100 ⁇ g/mL, GM3 inhibited the growth of normal human endothelial cells (4.5x10 4 on day 5, compared with 76x10 4 in controls.
  • US Pat. No. 5,330,977 describes n-deacetyl-lysoganglioside derivatives for use as phospholipase A2 inhibitors for the treatment of proliferative and autoimmune diseases, including various forms of cancer, psoriasis, and rheumatoid arthritis.
  • Anti-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 the inhibition of NF- ⁇ B activity (Kopp and Ghosh, Science, 265: 956-959 (1994); Ray and Prefontaine, Proc. Natl. Acad. ScL U.S.A., 91, 752-756 (1994)).
  • TNF- ⁇ production There are several drugs which show inhibitory activity for TNF- ⁇ production, including IL-4, IL-10 and transforming growth factor- ⁇ (TGF- ⁇ ), and clinical trials using antibody against TNF- ⁇ and soluble TNF- ⁇ receptor are underway.
  • synthetic compounds for example, new derivatives of 1-thiadibenzoazulene (US patent 6,897,211) and dibenzoazulene compounds (US patent 6,897,211 ) have been developed for their TNF- q inhibitory activity. Because these agents are TNF- ⁇ inhibitors, concerns have been raised about the risks of infection resulting from such an immunosuppression.
  • Inhibitors of NO production have focused on the development of agents that specifically inhibit the enzymatic activities of inducible NO synthase (iNOS), the enzyme responsible for NO synthesis.
  • iNOS inducible NO synthase
  • analogues of NO precursors such as L- arginine and L-citrulline, and derivatives of aminoguanidine and isothiourea are under investigation (Babu and Griffith, Current Opinion in Chemical Biology, 2, 491-500 (1998)).
  • Curcumin, capsaicin, caffeic acid derivatives, and some other natural compounds have been reported to inhibit the activation of NF- ⁇ B, while dexamethasone, a representative of the anti-inflammatory agent of the glucocorticoid family, inhibits NO production by inhibiting the expression of the iNOS gene.
  • dexamethasone a representative of the anti-inflammatory agent of the glucocorticoid family
  • the molecular mechanism of dexamethasone in the NO synthesis has been known to be the suppression of iNOS gene transcription by the way of the inhibition of NF- ⁇ B activation (De Vera et al., Am. J. Physiol., 273, 1290-1296 (1997)).
  • TGF- ⁇ s are a group of polypeptides consisting of at least 5 distinct but closely related members having considerable structural and biological homologies (Roberts, A. B., et al. In: Peptide Growth Factors and their Receptors Vol. 1 , pp.
  • TGF- ⁇ s are homodimeric proteins of about 25 kDa comprising identical 12.5 kDa polypeptide chains linked through disulphide bridges. They may form latent complexes with other proteins, and these complexes may be activated by acid treatment or mild proteolysis
  • Suitable methods are known for purifying TGF- ⁇ s from various species such as human, mouse, green monkey, pig, bovine, and chick, and from various cellular sources such as bone cells, platelets, or placental cells, as well as for producing it in recombinant cell cultures and for determining their activities (European Patents 200,341; 169,016; 268,561 and 267,463; G.B. Patent. Application 2,146,335; U.S. Patent 4,774,322).
  • TGF- ⁇ is multifunctional, as it can either stimulate or inhibit cell proliferation, differentiation, and other critical processes in cell functions (M. Sporn, Science, 233:532-534 (1986)). TGF- ⁇ has been shown to have numerous regulatory actions on a wide variety of cells. Recent studies indicate an important role for TGF- ⁇ in immune cells (J. Kehrl et al., J. Exp. Med., 163: 1037-1050 (1986); H-J. Ristow, Proc. Natl. Acad. Sci. U.S.A., 83: 5531-5533 (1986); A. Rook et al., J.
  • TGF- ⁇ has been described as a suppressor of cytokines production (interferon- ⁇ (IFN- ⁇ ), TGF- ⁇ ), indicating a possible use as an immunosuppressant for treating inflammatory disorders (European Patent 269,408; U.S. Patent 4,806,523). Further, TGF- ⁇ induces collagen secretion in human fibroblast cultures (Roberts et al., Proc. Nat. Acad. Sci. USA 83: 4167-4171 (1986)), stimulates the release of PGs and mobilization of calcium (A.
  • IFN- ⁇ interferon- ⁇
  • TGF- ⁇ induces collagen secretion in human fibroblast cultures (Roberts et al., Proc. Nat. Acad. Sci. USA 83: 4167-4171 (1986)), stimulates the release of PGs and mobilization of calcium (A.
  • TGF- ⁇ is produced by both immune and non-immune cells, and it exhibits a broad range of functions, including the modulation of immune responses.
  • Oral administration of TGF- ⁇ 2 is well known to decrease the inflammatory process.
  • Fell et al. ⁇ Aliment Pharmacol. Then 14 (3): 281-289, (2000) have demonstrated the antiinflammatory effect of a rich polymeric TGF- ⁇ 2 diet (CT3211 : Nestle Vevey, Switzerland) in children with IBD.
  • CT3211 rich polymeric TGF- ⁇ 2 diet
  • 29 children with IBD were treated with CT3211 during 8 weeks. Following the treatment, the results shown that 79% of the children were in complete remission from their disease.
  • TGF- ⁇ s include TGF- ⁇ 2
  • TGF- ⁇ 2 have a number of immunoregulatory properties and act at several stages of the 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 interleukins including IL-1 , IL-2 and IL-3 and other immunoregulatory agents such as TNF- ⁇ and interferons (Roberts, A. B. et al. , Supra).
  • TGF- ⁇ s interfere with certain accessory cell functions important in antigen presentation and they were specifically shown to suppress major histocompatibility complex (MHC)-Class Il expression by melanomas, glial cells and astrocytes (Czamiecki, 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)).
  • MHC-Class Il expression on epithelial cells in the intestine by TGF- ⁇ s or MGF has been reported in European patent EP0527283.
  • US patent 6,649,168 describes a pharmaceutical dry powder composition
  • a TGF- ⁇ in a water soluble salt (calcium chloride, calcium phosphate, potassium acetate, lithium acetate, ammonium acetate or ammonium bicarbonate) for therapeutic applications such as promotion and acceleration of wound healing, bone repair, soft tissue repair, nerve repair, stroke therapy, cancer treatment, bone marrow protection, cardioprotection, angiogenesis induction, mammalian cell growth regulation and induction of anti-inflammatory and immunosuppressive activity.
  • a water soluble salt calcium chloride, calcium phosphate, potassium acetate, lithium acetate, ammonium acetate or ammonium bicarbonate
  • autoimmune-related diseases such as multiple sclerosis (MS)
  • Those methods comprise administering IL-10 and TGF- ⁇ in combination to a person afflicted with or predisposed to an autoimmune disease.
  • these cytokines act in a synergistic manner as suppressor factors in order to inhibit the activation of self-reactive T cells that are involved in autoimmune diseases.
  • U.S. Pat. No. 5,461,033 describes the use of an acid casein fraction isolated from bovine milk and containing TGF- ⁇ 2 in the treatment of Crohn's disease.
  • the data reported in the patent is based upon in vitro experimentation and the patent gives no specific description of nutritional formulas that can be used in the treatment of Crohn's disease.
  • U.S. Pat. No. 5,952,295 gives a precise formulation of enteral preparation for the treatment of inflammatory conditions in the gastro-intestinal tract using a casein fraction with TGF- ⁇ 2 and reports improvement of such inflammatory conditions in an open-label study with 11 young patients.
  • European Patent 0313515 describes a milk growth factor fraction obtained from milk, and the use of this fraction for treating trauma, suppressing the immune response or treating cancer in mammals.
  • Patent application WO 02/05828 proposes the treatment of skin disorders by neutral or basic growth factors from milk or colostrum.
  • Japanese Patent 14308090 describes a fraction from colostrum with TGF- ⁇ activity.
  • a dairy growth factors fraction obtained from chromatographic methods, especially with an ionic exchange step, is described in US patent 6,447,808. This fraction can be used for the treatment of gastrointestinal injuries, ulcers, or surface wounds.
  • Patent application WO 01/25276 also describes isolation of growth factors from dairy products by the use of chromatography. Different fractions can be produced by this process, especially fractions enriched in TGF- ⁇ and IGF-1.
  • Patent application WO 03/006500 describes a process to obtain a dairy fraction enriched in TGF- ⁇ . This process is based on a precipitation step 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 ⁇ g of TGF- ⁇ per gram of proteins. This fraction is proposed for the treatment of auto-immune diseases, arthritis, osteoporosis, wound healing and graft rejection.
  • One aim of the present application is to provide a new and simple method for obtaining a dairy fraction enriched in TGF- ⁇ that can be used as an effective chronic intestinal inflammatory disorders treatment, a composition containing such a fraction and uses thereof.
  • a dairy-derived composition for prophylaxis or treatment of an inflammatory disease or a related disorder in a subject wherein said composition comprises TGF- ⁇ 1 , TGF- ⁇ 2, IGF-1, beta-lactoglobulin, and at least 15% (w/w) of whey derived proteins.
  • the concentration of TGF- ⁇ 1 is between about 0.1 and 5 ⁇ g per gram of composition, and more preferably between about 0.2 and 1.2 ⁇ g per gram of composition.
  • the concentration of TGF- ⁇ 2 is preferably between about 5 and 50 ⁇ g per gram of composition, and more preferably between about 10 and 18 ⁇ g per gram of composition.
  • the concentration of whey derived proteins is at least 50% (w/w), and more preferably at least 80% (w/w).
  • composition may further comprise between 0.1 and 15% of minerals.
  • the composition comprises at least 35%, and preferably at least 70% of water-soluble proteins.
  • the inflammatory disease may be for example chronic inflammatory disease, or chronic intestinal inflammatory disease, such as Crohn's disease or ulcerative colitis.
  • the subject can be an animal or a human.
  • a dairy-derived composition comprising TGF- ⁇ 1, TGF- ⁇ 2, IGF-1 , beta-lactoglobulin and at least 15% (w/w) of whey derived proteins for prophylaxis or treatment of inflammatory diseases or related disorders.
  • 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- ⁇ 1, TGF- ⁇ 2, IGF-1, beta-lactoglobulin, and at least 15% (w/w) of whey derived proteins.
  • chronic inflammatory diseases includes rheumatoid arthritis, psoriasis, cutaneous inflammation, atopic dermatitis, encephalitis, hypersensitivity pneumonitis, chronic lung inflammation, ischemia- reperfusion injury, systemic lupus erythematosus, myositis, ankylosing spondylitis, scleroderma, acute inflammatory demyelinating polyradiculoneuropathy, vasculitis, appendicitis, arachnoiditis, myocarditis, acute cholecystitis, chronic airflow obstruction, chronic hepatitis, chronic obstructive pulmonary disease, conjunctivitis, dermatitis, enteritis, gingivitis, hepatitis, ileitis, asthma, and/or intestinal chronic inflammation diseases such as Crohn's disease
  • one aspect of the present application contemplates a method for reducing the effects of a proliferative and/or inflammatory disorder in a subject, the method comprising treating a chronic inflammatory disease with an effective amount of a composition capable of preventing or treating chronic inflammatory diseases and related disorder, as described herein.
  • the term "inflammation” as used herein is intended to represent the normal response of the immune system to infection or irritation.
  • the present application only addresses the cellular compartment of the immune response, also referred to as the Th1 component of the immune system, and is not addressing the humoral, or exudative portion of the immune response, also referred to as the Th2 component of the immune system.
  • inflammatory disease and "inflammatory disorders” as used herein are intended to mean any disease or disorder affecting the normal response of the immune system.
  • the disease or disorder can involve either an excessive or an insufficient response of the immune system to infection or irritation, a lack of response following infection or irritation, and a response in the absence of infection or irritation.
  • Such disorders can be for example, without limitation, ulcerative proctitis, proctosigmoiditis, left-sided colitis, pan-ulcerative (total) colitis, psoriatic arthritis, ankylosing spondylitis, juvenile ankylosing spondylitis, seronegative enthesopathy, arthropathy syndrome,
  • inflamed tissue refers to a tissue affected by inflammation and affected cells contained within the tissue. Cells that have migrated to the inflammation site, such as leukocytes, are excluded from the definition of this term.
  • the inflamed tissue is from a mammalian species, preferably a human.
  • an effective amount and "therapeutically effective amount” as used herein are intended to mean an amount sufficient to initiate a beneficial or desired clinical result, such as an improvement of the condition of the patient.
  • An effective amount can be administered in one or more doses.
  • 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. One skilled in the art can easily and without difficulty monitor the inflamed tissue so as to determine what will be such effective amount.
  • mammals include, but are not limited to, farm animals, sport animals, rodents, primates, and pets.
  • minerals as used herein is intended to mean salt constituents and minerals normally found in milk.
  • calcium, phosphorus, magnesium, potassium, sodium, chloride, sulfur, and citric acid can be used in various concentrations. This also means that trace elements known in the art as being present in milk can be assimilated or inferred therein.
  • treatment is intended to mean a medical remedy to improve or alleviate in part or totally the condition of a subject suffering from inflammation or inflammatory disorders.
  • prophylaxis as used herein is intended to mean any procedure directed to the prevention of inflammation or inflammatory disorders. It will be understood that this term represents both primary prophylaxis, i.e. prevention of the apparition of the disease or disorder, and secondary prophylaxis, i.e. prevention of the worsening of an already developed disease or disorder.
  • Fig. 1 shows , a dose-dependent inhibition of ConA-induced murine splenocytes proliferation by the composition of the present application
  • Fig. 2 shows a dose-dependent inhibition of ConA-induced murine splenocytes IL-2 production by the composition produced in example 1 and described in example 2;
  • Fig. 3 shows a dose-dependent inhibition of ConA-induced murine splenocytes IFN ⁇ y production by the composition produced in example 1 and described in example 2;
  • Fig. 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), a chemical inflammatory inducer, as described in detail in example 4;
  • TNBS 2,4,6-trinitrobenzenesulfonic acid
  • Fig. 5 illustrates the effects of colorectal administrations of the composition produced in example 1 and described in example 2 on weight-to-length ratios of the colons of Sprague-Dawley rats treated with TNBS, as described in detail in example 4;
  • Fig. 6 illustrates the effects of colorectal administrations of the composition produced in example 1 and described in example 2 on macroscopic damages of rat colon tissues previously irritated with TNBS, as described in detail in example 4;
  • Fig. 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;
  • Fig. 8 shows hematocrit levels before and during oral treatment with different doses of the composition produced in example 1 and described in example 2, where 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
  • Fig. 9 shows hemoglobin levels before and during oral treatment with different doses of the composition produced in example 1 and described in example 2, where 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).
  • compositions and a method for treating chronic inflammatory diseases in an individual wherein the composition effective in stimulating a specific immunological response against a physiological imbalance aberrantly expressed in inflamed tissues.
  • This composition comprises a cocktail of products that shares growth and regeneration characteristics of a growth factor that is aberrantly expressed in inflamed tissues (such as human inflamed tissues).
  • Particular growth factors included in the composition of the present application are, but not limited to, TGFs, such as TGF- ⁇ 1 and TGF- ⁇ 2, beta- lactoglobulin and at least 15% of dairy derived proteins.
  • the growth factors present in the composition are distinguished by having a strong anti-inflammatory action.
  • concentration of the compounds in the composition of the present application may be adjusted depending of the needs.
  • TGF- ⁇ 1 can be present at concentration of between 0.1 to 5 ⁇ g/g of composition and TGF- ⁇ 2 can be present at concentration of between 5 and 50 ⁇ g/g of composition.
  • TGF- ⁇ 1 is present at concentration of 0.2 to 1.2 ⁇ g/g, and TGF- ⁇ 2 at between 10 to 18 ⁇ g/g of composition.
  • the preparation can be used daily in a customary manner.
  • the composition comprises at least 70% (w/w) of whey derived proteins.
  • the total concentration of proteins in the composition would normally be of at least 80%.
  • Fat concentration can be generally found at concentration between about 0.5 to 10 % (w/w) in the composition.
  • the proteins are water-soluble.
  • the proteins may be comprised of at least 60% (w/w) of ⁇ -lactoglobulin, and/or between 0.1 to 30% (w/w) of lactoferrin.
  • the active compounds according to the application can be mixed with customary pharmaceutically tolerable diluents or vehicles and, if appropriate, with other auxiliaries 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. It can also be administered as suppositories, vaginal suppositories, and/or parenterally, e.g. in the form of solutions, emulsions, creams or suspensions.
  • Preparations to be administered orally can contain one or more additives such as sweeteners, aromatizing agents, colorants and preservatives.
  • Tablets can contain the composition mixed with customary pharmaceutically tolerable auxiliaries, as for example inert diluents such as calcium carbonate, sodium carbonate, lactose and talc; granulating agents and agents which promote the disintegration of the tablets on 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 extending 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 optionally to use organic solvents as auxiliary solvents.
  • tablets can also contain additives, such as sodium citrate, calcium carbonate and dicalcium phosphate together with other various additives, such as starch, preferably potato starch, gelatin and the like.
  • additives such as sodium citrate, calcium carbonate and dicalcium phosphate together with other various additives, such as starch, preferably potato starch, gelatin and the like.
  • lubricants such as magnesium stearate, sodium lauryl sulphate and talc can additionally be used for tableting.
  • the active compounds can be mixed, apart from with the above-mentioned auxiliaries, with various flavors enhancers or colorants.
  • solutions of the composition using suitable liquid excipients can be employed.
  • Capsules can contain the composition as a single constituent or mixed with a solid diluent such as calcium carbonate, calcium phosphate or kaolin.
  • the injectable formulations are also formulated in a manner known perse.
  • antioxidants can be added to the composition and to the pharmaceutical and topical formulations.
  • the use of natural or naturally identical compounds such as, for example, tocopherols is particularly preferred here.
  • the antioxidants mentioned are contained in the compositions according to the application, 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 or alcoholic solutions, aqueous suspensions or emulsions.
  • chronic inflammatory diseases vary according to each specific disease. For example, intestinal inflammatory diseases are considered to affect 0.3% of American population.
  • the composition can be used prophylatically and for the treatment of the disorders.
  • the composition is administered in order to decrease manifestations of the disease in frequency and strength.
  • Treatment in the manifest stage leads to its curtailment and to the alleviation of the symptoms.
  • composition can also be administered colorectally or protected with enteric coating for delivery in the intestinal part of digestive tractus.
  • the composition can also be incorporated into customary oral or pharmaceutical bases 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.
  • the composition also addresses the underlying T-cell disorder that results in an inflammatory condition.
  • the Applicants have recognized that most, if not all, of the current therapies in the prior art for T-cell mediated inflammatory conditions are designed to eliminate T-cells and to thereby improve the inflammation condition.
  • a major problem with those treatments is that there is a possibility that the therapy itself is so toxic that it may promote recurrence during healing. The toxicity of these treatments can unleash some or all of the cytokines that are associated with the promulgation of these chronic and often rebounding diseases.
  • the synergy of the two TGF- ⁇ factors of the composition results in a non-toxic, highly effective treatment for chronic inflammatory diseases that is without the side effects observed with virtually all other therapies for those diseases.
  • composition of the ingredient is presented in table 1. Proteins were analyzed by the Dumas method, while fat was analyzed by the Mojonnier method, humidity by drying in a dry oven, beta-lactoglobuline by RP-HPLC and growth factors by ELISA methods.
  • TGF- ⁇ 2 11.700 pg/mg
  • TGF- ⁇ 1 300 pg/mg EXAMPLE 3 Inhibition of proliferation of mice splenocytes
  • TGF- ⁇ has multiple immunosuppressive effects at the cellular level and has been described as inhibiting type 1 and type 2 cells, B cells, CD8 + T cells, macrophages and natural killer cells.
  • TGF- ⁇ blocks cell-cycle progression and may have a direct effect on expression of the gene encoding IL-2 (Brabletz et al., MoI. CeI. Biol., 13(2): 1155-1162, 1993). Importantly, it can suppress the expression of IL-12 and IL-2 receptors, as well as downregulate MHC class Il expression on macrophages (specifically by antagonizing TNF- ⁇ and IFN- ⁇ ) (Harber et al., Exp. Rev. MoI.
  • TGF- ⁇ - knockout mice who incur severe, spontaneous, multi-organ, autoimmune disease associated with autoantibody production, which proves fatal in early life (Harber et al., Exp. Rev. MoI. Med., 27 Nov., http://www-ermm.cbcu.cam.ac.uk/00002143h.htm, 2000).
  • T lymphocytes isolated from murine spleen are immune cells that respond to ConA (mitogenic stress) by proliferation and cytokine production (Mercier, et al. Internal Dairy J., 14: 175-183, (2004); Wong, et al. J. Dairy ScL, 81: 1825-1832, (1998)).
  • ConA mitogenic stress
  • cytokine production Mercier, et al. Internal Dairy J., 14: 175-183, (2004); Wong, et al. J. Dairy ScL, 81: 1825-1832, (1998).
  • the purpose of this example was to evaluate the inhibition effect of the composition produced in example 1 and described in example 2 on splenocytes proliferation and production of proinflammatory cytokines.
  • mice Female BALB/c mice, 6-8 weeks old, were obtained from Charles River (Montreal,
  • mice were sacrificed by CO 2 inhalation and single-cell suspensions were prepared individually from murine spleen under aseptic conditions.
  • Murine lymphocytes (1.25 x 10 6 cells/ml) were treated with ConA (1.25 ⁇ g/ml) for 72 hours.
  • Alamar blue® was added 24 hours before measuring ConA-induced lymphocytes proliferation.
  • Cytokines (IL-2 & IFN ⁇ y) levels were also determined in the supernatant following the incubation period.
  • BSA was used as a protein control (1-1000 ⁇ g/ml).
  • composition produced in example 1 and described in example 2 (1, 10, 50, 100, 500, and 1000 ⁇ g of the total composition per ml of culture medium), was added to the cells for the whole incubation period and dose-dependently decreased 1) the lymphocyte proliferation (Fig. 1), 2) the production of IL-2 (Fig. 2), and 3) the production of IFN- ⁇ (Fig. 3).
  • Fig. 1 lymphocyte proliferation
  • Fig. 2 lymphocyte proliferation
  • Fig. 2 Fig. 2
  • IFN- ⁇ Fig. 3
  • EXAMPLE 4 Test on TNBS-induced inflammation in healthy rats
  • Inflammatory bowel diseases is a term used to describe a collection of diseases that involve the intestine and are characterized by chronic inflammation, sometimes accompanied by ulceration in the small or large intestine.
  • Cytokines play an important role in modulating the immune system since they are rapidly synthesized and secreted from activated inflammatory cells and induce the production of adhesion molecules and 0 other inflammatory mediators, such as reactive oxygen metabolites, nitric oxide, and lipid mediators (De Winter et al., Am J Physiol 276: G1317-1321 , 1999). Cytokines induce, amplify, prolong, and inhibit inflammation.
  • mice Male Sprague Dawley (Charles River Laboratories, Montreal), were kept in individual cages, at 20 0 C and 55% relative humidity with 12h light/12-h dark cycles, in a facility o meeting the Canadian Council on Animal Care guidelines.
  • the rats were subjected to a 24-hour starving period prior to inflammatory stress induction. Colonic inflammation was induced by using a technique from Togawa et al. (Am. J. Physiol., 283:187-195, 2002). Briefly, the rats were lightly anesthetised with isoflurane after overnight food deprivation, and a polyethylene catheter was inserted 8 cm into the colon via the anus.
  • TNBS (Sigma-Aldrich, Canada) dissolved in 50% (vol/vol) aqueous ethanol (25mg/ml) was 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.
  • Body weight As shown in Fig. 4, body weights of the animal increased constantly prior to fasting and TNBS injection in all groups. Following PBS injection (for control animals only), animals regained the weight lost during the fasting period. In TNBS-treated animals, however, the animals did not regain the body weight lost during the fasting period. As for the animals treated with the composition produced in example 1 and described in example 2, body weight was increased following the TNBS injection. Weight-to-length ratio. TNBS injection increased weight-to-length ratio in colon for both TNBS and the composition produced in example 1 and described in example 2 groups, indicating the presence of inflammation and edema. However, the increase was significantly (P ⁇ 0.05) inferior in animals treated with the composition produced in example 1 and described in example 2 than in TNBS-treated animals (Fig. 5).
  • TNBS injection increased macroscopic scores 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.
  • the increase was significantly inferior (P ⁇ 0.05) in animals treated with the composition produced in example 1 and described in example 2 than in TNBS-treated animals (Fig. 6 and Fig. 7).
  • HLA-B27 transgenic rat expressing HLA-B27 and human ⁇ 2m, exhibits a phenotype similar to humans suffering B27 related rheumatic disorders such as reactive arthritis, ankylosing spondylitis, psoriasis and inflammatory bowel disease.
  • 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 primary goal 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 HLA-B27 rat model.
  • IBD i.e. positive HemoccultTM
  • the composition produced in example 1 and described in example 2 (7.82, 78.2 and 782 mg/kg) was orally administered daily via a gastric needle (total volume of 1 ,5 ml).
  • Hematocrits As shown is Fig. 8, hematocrit levels were higher 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 the value observed before the beginning of 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 to 782 mg/kg of the composition produced in example 1 and described in example 2 (Fig. 9).

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Developmental Biology & Embryology (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Rheumatology (AREA)
  • Virology (AREA)
  • Pain & Pain Management (AREA)
  • Diabetes (AREA)
  • Molecular Biology (AREA)
  • Endocrinology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

La présente invention concerne une composition dérivée du lait ainsi que des méthodes associées ou des utilisations de ladite composition dans la prophylaxie et le traitement de maladies inflammatoires, notamment de l'inflammation intestinale chronique, de l'inflammation et de troubles associés. La composition comprend des TGF-ß1, des TGF-ß2, des IGF-1 et de la bêta-lactoglobuline et présente une concentration protéique d'au moins 15 % (p/p) de protéines dérivées du lactosérum.
PCT/CA2006/001637 2004-12-09 2006-10-04 Composition derivee du lait enrichie en tgf-$g(b) pour le traitement de l'inflammation WO2007038870A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
NZ567126A NZ567126A (en) 2005-10-04 2006-10-04 A dairy derived composition comprising beta-lactoglobulin, IGF and TGF-beta for use in treating chronic intestinal inflammatory disease
US11/695,757 US7763257B2 (en) 2004-12-09 2007-04-03 Compositions comprising transforming growth factor (TGF)-β1 and TGF-β2 in admixture of proteins obtained from dairy products

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US72296905P 2005-10-04 2005-10-04
US60/722,969 2005-10-04

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US11/007,912 Continuation-In-Part US20060127493A1 (en) 2004-12-09 2004-12-09 Composition for treating psoriasis
US11/695,757 Continuation-In-Part US7763257B2 (en) 2004-12-09 2007-04-03 Compositions comprising transforming growth factor (TGF)-β1 and TGF-β2 in admixture of proteins obtained from dairy products

Publications (1)

Publication Number Publication Date
WO2007038870A1 true WO2007038870A1 (fr) 2007-04-12

Family

ID=37905941

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CA2006/001637 WO2007038870A1 (fr) 2004-12-09 2006-10-04 Composition derivee du lait enrichie en tgf-$g(b) pour le traitement de l'inflammation

Country Status (2)

Country Link
NZ (1) NZ567126A (fr)
WO (1) WO2007038870A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008119185A1 (fr) * 2007-04-03 2008-10-09 Advitech Inc. Procédé de fabrication d'une composition anti-inflammatoire à base de produit laitier
WO2008151449A1 (fr) * 2007-06-11 2008-12-18 Advitech Inc. Procédé de traitement d'eczéma atopique
US8937043B2 (en) 2001-07-13 2015-01-20 Piere Jouan Biotechnologies S.A. Method for obtaining a TGF-beta enriched protein fraction in activated form, protein fraction and therapeutic applications
WO2021070183A1 (fr) * 2019-10-07 2021-04-15 Maolac Ltd. Composition pour renforcer le système immunitaire
WO2023079080A1 (fr) 2021-11-05 2023-05-11 Frieslandcampina Nederland B.V. Utilisation de tgf dans la prévention d'une infection virale des voies respiratoires

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040097714A1 (en) * 2001-07-13 2004-05-20 Jean-Louis Maubois Method for obtaining a tgf-beta enriched protein fraction in activated form protein fraction and therapeutic applications
CA2489851A1 (fr) * 2004-12-09 2006-06-09 Yves Pouliot Composition pour traiter le psoriasis

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040097714A1 (en) * 2001-07-13 2004-05-20 Jean-Louis Maubois Method for obtaining a tgf-beta enriched protein fraction in activated form protein fraction and therapeutic applications
CA2489851A1 (fr) * 2004-12-09 2006-06-09 Yves Pouliot Composition pour traiter le psoriasis

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
PLAYFORD R.J. ET AL.: "Colostrum and milk-derived peptide growth factors for the treatment of gastrointestinal disorders", AM. J. CLIN. NUTR., vol. 72, no. 1, July 2000 (2000-07-01), pages 5 - 14, XP000971624 *
POULIN Y. ET AL.: "Evaluation of the safety and efficacy of XP-828L in the treatment of plaque psoriasis: An open-label study", 63RD ANNU. MEET. AM. ACAD. DERMATOL., NEW ORLEANS, 18 February 2005 (2005-02-18), pages COVER PAGE + ABSTR. NO. P2830 *
POULIN Y. ET AL.: "Safety and efficacy of a milk-derived extract in the treatment of plaque psoriasis: an open-label study", J. CUTAN. MED. SURG., vol. 9, no. 6, December 2005 (2005-12-01), pages 271 - 275, XP003011247 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8937043B2 (en) 2001-07-13 2015-01-20 Piere Jouan Biotechnologies S.A. Method for obtaining a TGF-beta enriched protein fraction in activated form, protein fraction and therapeutic applications
WO2008119185A1 (fr) * 2007-04-03 2008-10-09 Advitech Inc. Procédé de fabrication d'une composition anti-inflammatoire à base de produit laitier
WO2008151449A1 (fr) * 2007-06-11 2008-12-18 Advitech Inc. Procédé de traitement d'eczéma atopique
WO2021070183A1 (fr) * 2019-10-07 2021-04-15 Maolac Ltd. Composition pour renforcer le système immunitaire
CN114585372A (zh) * 2019-10-07 2022-06-03 毛拉克有限公司 用于增强免疫系统的组合物
WO2023079080A1 (fr) 2021-11-05 2023-05-11 Frieslandcampina Nederland B.V. Utilisation de tgf dans la prévention d'une infection virale des voies respiratoires

Also Published As

Publication number Publication date
NZ567126A (en) 2010-09-30

Similar Documents

Publication Publication Date Title
US6852700B1 (en) Colostrinin, and uses thereof
AU2006203622A1 (en) Inflammation-inhibiting compounds
Chu et al. Oral fungal immunomodulatory protein-Flammulina velutipes has influence on pulmonary inflammatory process and potential treatment for allergic airway disease: A mouse model
WO2007038870A1 (fr) Composition derivee du lait enrichie en tgf-$g(b) pour le traitement de l'inflammation
RU2723097C1 (ru) Способ получения яичного желтка с высоким содержанием af-16
JP2018531214A6 (ja) 高含量のａｆ−１６を有する卵黄の産生方法
EP0977581B1 (fr) Composition contenant des histones et utile dans le traitement de l'arthrite rhumatoide
US20100272708A1 (en) Method for the treatment of type and type iv hypersensitivity
EP0655925B1 (fr) Compositions therapeutiques anti-cancereuses contenant un concentre de proteines de petit-lait
US7763257B2 (en) Compositions comprising transforming growth factor (TGF)-β1 and TGF-β2 in admixture of proteins obtained from dairy products
MX2008004479A (es) Composiciones para tratar la inflamacion
US20120077747A1 (en) Fish-derived protein lysate, and uses thereof as immunomodulatory and/or anti-inflammatory agent
US9138454B2 (en) Transferrin for use in the treatment and/or prophylaxis of autoimmune diseases
EP2144623B1 (fr) Procédé de fabrication d'une composition anti-inflammatoire à base de produit laitier
US20110098216A1 (en) Therapeutic uses of colostrinin
US20030007964A1 (en) Method for treating rheumatoid arthritis with composition containing histone
WO2016011133A1 (fr) Traitement peptidique pour empêcher la résistance à l'insuline et les diabètes de type 2
CA2545726A1 (fr) Utilisation de l'interferon tau pour traiter l'obesite et favoriser la perte de poids
Petrov Myelopeptides
WO2011113100A1 (fr) Méthode de traitement d'un trouble osseux
Crabtree et al. T lymphocyte stimulation of human small intestinal glycoprotein biosynthesis: effects of anti-CD3 antibody on normal and coeliac mucosa
KR20090096688A (ko) 자가면역 질환의 예방과 치료를 위한 약제의 제조에서 티모신 알파 1의 용도

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 11695757

Country of ref document: US

121 Ep: the epo has been informed by wipo that ep was designated in this application
WWP Wipo information: published in national office

Ref document number: 11695757

Country of ref document: US

WWE Wipo information: entry into national phase

Ref document number: 567126

Country of ref document: NZ

WWE Wipo information: entry into national phase

Ref document number: MX/a/2008/004479

Country of ref document: MX

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 06790797

Country of ref document: EP

Kind code of ref document: A1