Patent Application
20150306143
The present invention relates to the field of treatment of an inflammatory autoimmune condition. The present invention relates in particular to methods for treating an inflammatory autoimmune condition and immune related diseases, using a medicament comprising human Tr1 cells directed against a human heat shock protein.
Inflammatory autoimmune diseases affect millions of people worldwide and can have devastating effects on lifespan and quality of life.
Conventional therapies for treating inflammatory autoimmune diseases in humans are based on non-specific immunosuppression, such as that mediated by steroids or cytotoxic agents. More recent strategies include the blockade of several pro-inflammatory cytokine pathways, such as those that involve TNF and IL-1. However, none of these strategies reverses the pro-inflammatory process or cures the disease. When the treatment is stopped, a reactivation of inflammation occurs within months, thereby prompting life-long treatment with immunosuppressive agents. This inevitably leads to serious side effects. Therefore, there is a need for novel treatments for inflammatory autoimmune diseases. Various studies have demonstrated that heat shock proteins (HSPs), and in particular HSP60 and HSP70, constitute a group of autoantigen involved in the regulation of inflammatory autoimmune diseases.
Therapies targeting HSPs for treating inflammatory autoimmune diseases have therefore been proposed such as immunization with E. coli lysate or peptides derived from HSP60 or use of DNA vaccines comprising a nucleic acid sequence encoding HSP60, HSP70 or HSP90 or fragment thereof.
In the present invention, the Applicant aims to provide an alternative treatment for an inflammatory autoimmune condition based on the use of Tr1 cells directed against a human HSP.
An object of the invention is a medicament comprising at least one human Tr1 cell population directed against a human HSP. Another object of the invention is a pharmaceutical composition comprising at least one human Tr1 cell population directed against a human HSP in combination with one or more pharmaceutically acceptable carriers.
Another object of the invention is the use of a composition comprising at least one human Tr1 cell population directed against a human HSP for the preparation of a medicament or a pharmaceutical composition for treating an inflammatory autoimmune condition.
According to an embodiment of the invention, said inflammatory autoimmune condition is an arthritis condition, preferably rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis and juvenile idiopathic arthritis. According to another embodiment, said inflammatory autoimmune condition is a multiple sclerosis condition. According to another embodiment, said inflammatory autoimmune condition is an intestinal inflammatory condition, preferably Crohn's disease and ulcerative colitis. According to another embodiment, said inflammatory autoimmune condition is a vasculitis, preferably Wegener's disease and atherosclerosis. According to another embodiment, said inflammatory autoimmune condition is asthma. According to another embodiment, said inflammatory autoimmune condition is transplant rejection or graft versus host disease. According to another embodiment, said inflammatory autoimmune condition is an inflammatory condition of the biliary duct, preferably primary biliary cirrhosis and primary sclerosing cholangitis.
According to another embodiment, the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with one or more therapeutic agents used for treating an inflammatory autoimmune condition.
According to another embodiment, the administration to said subject of an effective amount of the medicament or the pharmaceutical composition is in combination with one or more medicaments or pharmaceutical compositions comprising at least one Tr1 cell population or clone directed against an antigen known to be involved in the inflammatory autoimmune condition.
Another object of the invention is a process for treating an inflammatory autoimmune condition in a subject in need thereof, said process comprising the steps of:
IL-10, IL-4 and IFNγ secretion of HSP60 specific Tr1 cell clones were measured in 48 hours supernatant of anti-CD3+anti-CD28 monoclonal antibodies activated cells.
The suppressive activity of HSP60 Tr1 clones was assessed by evaluating the inhibitory action of their supernatants on autologous CD4+ T lymphocytes proliferation. CD4+ T lymphocytes were activated using anti-CD3+anti-CD28 monoclonal antibodies and Interleukin-2 in the presence or absence of the supernatants of activated Tr1 clones. Cell proliferation of the autologous CD4+ T cells was measured after five days. Results show that the addition of Tr1 cell's supernatants to CD4+ T lymphocytes inhibits T-cell proliferation.
The term “Tr1 cells” as used herein refers to cells having the following phenotype at rest CD4+CD25−FoxP3− and capable of secreting high levels of IL-10 and significant levels TGF-β upon □activation. Tr1 cells are characterized, in part, by their unique cytokine profile: they produce high levels of IL-10, significant levels of TGF-β and intermediate levels of IFN-γ, but little or no IL-4 or IL-2. The cytokine production is typically evaluated in cultures of cells after activation with polyclonal activators of T lymphocytes such as anti-CD3+ anti-CD28 antibodies or Interleukin-2, PMA+ionomycin. Alternatively, the cytokine production is evaluated in cultures of cells after activation with the specific T-cell antigen presented by antigen presenting cells. High levels of IL-10 correspond to at least about 500 pg/ml, typically greater than about 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, or 20 thousand pg/ml or more. Significant levels of TGF-β correspond to at least about 100 pg/ml, typically greater than about 200, 300, 400, 600, 800, or 1000 pg/ml or more. Intermediate levels of IFN-γ correspond to concentrations comprised between 0 pg/ml and at least 400 pg/ml, typically greater than about 600, 800, 1000, 1200, 1400, 1600, 1800, or 2000 pg/ml or more. Little or no IL-4 or IL-2 corresponds to less than about 500 pg/ml, preferably less than about 250, 100, 75, or 50 pg/ml, or less.
The term “antigen” as used herein refers to a protein, or peptide for which the cells of this invention are being used to modulate, or for use in any of the methods of this invention. In one embodiment, the term “antigen” may refer to a synthetically derived molecule, or a naturally derived molecule, which shares sequence homology with an antigen of interest, or structural homology with an antigen of interest, or a combination thereof. In one embodiment, the antigen may be a mimetope. A “fragment” of the antigen refers to any subset of the antigen, as a shorter peptide. A “variant” of the antigen refers to a molecule substantially similar to either the entire antigen or a fragment thereof. Variant antigens may be conveniently prepared by direct chemical synthesis of the variant peptide, using methods well-known in the art.
The term “subject” as used herein refers to a human being.
The term “effective amount” as used herein refers to an amount sufficient to cause a beneficial or desired clinical result (e.g. improvement in clinical condition).
The term “clone” or “clone population” as used herein refers to a population of differentiated cells being derived from a unique differentiated cell.
The term “treatment” or “treating” as used herein generally refers to a clinical intervention in an attempt to alter the natural course of the individual being treated, and may be performed during the course of clinical pathology. Desirable effects include, but are not limited to, alleviating symptoms, suppressing, diminishing or inhibiting any direct or indirect pathological consequences of the disease, lowering the rate of disease progression, ameliorating or palliating the disease state, and causing remission or improved prognosis. In the context of the invention, it refers to any improvement in the clinical symptoms of the autoimmune condition, as well as any improvement in the well being of the patients.
The term “autoimmune disease” as used herein refers to an immune response directed against an autoantigen.
The present invention relates to a method for treating an inflammatory autoimmune condition in a subject in need thereof, comprising the administration to said subject of a composition comprising at least one human Tr1 cell population directed against a human HSP.
Another object of the present invention is to provide a medicament comprising at least one human Tr1 cell population directed against a human HSP.
The present invention also intends to provide a pharmaceutical composition comprising at least one human Tr1 cell population directed against a human HSP in combination with one or more pharmaceutically acceptable carriers.
According to the invention, the “human Tr1 cell population” corresponds to Tr1 cells as described here above in the definitions and does not include CD4+CD25+ regulatory T cells or FoxP3+ regulatory T cells (natural or conventional Treg), TGF-β secreting Th3 cells, or regulatory NKT cells.
According to the invention, the term “human HSP” refers to human HSP10, HSP40, HSP60, HSP70, HSP90 and HSP100 families, fragments and mixtures thereof.
In a preferred embodiment of the invention, said Tr1 cells are directed against HSP60, HSP70, HSP90, fragments and mixtures thereof.
In a more preferred embodiment of the invention, said Tr1 cells are directed against HSP60 and fragments thereof.
Without wishing to be bound to a theory, the Applicant assumes that the injected Tr1 cell population directed against a human HSP would be activated in vivo by the HSPs whose expression is upregulated in an inflammatory autoimmune condition. Said activation of HSP specific Tr1 cells would lead to production of IL-10, which allows the suppression of CD4+ T cells.
In one embodiment of the invention, human Tr1 cells may be obtained by
In step b), IL-10 is present from 50 to 250 U/ml, preferably at 100 U/ml in the culture medium. Said method for obtaining Tr1 cells is described in Wakkach et al (Immunity 2003 May; 18(5):605-17).
Said method may also be carried out using dexamethasone and vitamin D3, or tolerogenised or immature DCs instead of the DCs of step b).
In another embodiment of the present invention, human Tr1 cells may be obtained by:
IFN-α is preferably present in the media at 5 ng/ml. In the step a), the media may further comprise an appropriate amount of IL-10, preferably at 100 U/ml.
In step b), the Tr1 cell population is cultured in a media comprising IL-15 to allow proliferation, IL-15 being preferably at 5 ng/ml in the media. Said method for obtaining Tr1 cells is described in the U.S. Pat. No. 6,746,670.
In still another embodiment of the invention, human Tr1 cells may be obtained by:
Preferably, the artificial antigen presenting cells express a HLA II system molecule and a human LFA-3 molecule and do not express the co-stimulation molecules B7-1, B7-2, B7-H1, CD40, CD23 and ICAM-1.
Said process, for obtaining Tr1 cells is described in the patent application WO02/092793.
In still another embodiment of the invention, human Tr1 cells may be obtained by:
Preferably, IL-10 is present in the media at 100 U/ml. Said method is described in Groux et al. (Nature 1997, 389(6652):737-42).
In still another embodiment of the invention, human Tr1 cells may be obtained by:
Leukocytes encompass several types of cells, which are characterized by their importance, their distribution, their number, their lifetime and their potentiality. These types are the following: the polynuclear or granular leukocytes, among which one finds the eosinophilic, the neutrophilic and the basophilic leukocytes, and the mononuclear cells, or peripheral blood mononuclear cells (PBMCs), which are large white blood cells and consist in the major cell types of the immune system (lymphocytes and monocytes). The leukocytes or the PBMCs can be separated from the peripheral blood by any method known to those skilled in the art. Advantageously, for the separation of the PBMCs, centrifugation may be used, preferably density gradient centrifugation, preferably discontinuous density gradient centrifugation. An alternative is the use of specific monoclonal antibodies. In certain embodiments PBMC are typically isolated from the whole blood product by means of Ficoll-Hypaque, using standard procedures. In other embodiments the PBMCs are recovered by means of leukapheresis.
Said method is described in the patent application WO2007/010406.
In still another embodiment, human Tr1 cells may be obtained by:
Said method can also be carried out with naïve or memory T cells instead of PBMC or leukocytes.
The Tr1 cell population thus obtained may further be expanded by culture in presence of cytokines such as Interleukin-2 and Interleukin-4. Alternatively, Interleukin-15 and Interleukin-13 could also be used in Tr1 cell expansion cultures.
In the methods described above, human Tr1 cells can be characterized by the identification method described in WO2005/000344. Said identification method of Tr1 cells is based on the detection of the simultaneous presence of expression products of genes coding CD4 molecule and molecules from the group comprising CD18 and/or CD11a, and CD49b. Tr1 cells can be identified and/or purified by Elisa, flow cytometry, or immunoaffinity methods with antibodies directed against said markers.
Tr1 cells can also be enriched by positive selection or negative selection using flow cytometry or magnetic beads. Such methods are also described in WO2005/000344.
In another embodiment of the present invention, the Tr1 cells directed to a joint-associated antigen may be expanded by the in vitro method described in WO2006/108882. Said method comprises:
Examples of factors which interact with the above mentioned cell surface proteins include:
In a preferred embodiment of the present invention, said Tr1 cells directed against a human HSP may be cloned by using conventional methods for cloning T cells.
In a preferred embodiment of the present invention, said composition comprising at least one human Tr1 cell population directed against a human HSP or at least one clone of human Tr1 cell directed against a human HSP may be frozen to be stored.
According to a preferred embodiment, said human Tr1 cell population is a human Tr1 clone population.
The pharmaceutically acceptable carriers useful herein are conventional. Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980) describes composition and formulations suitable for pharmaceutical delivery of the composition of the present invention. In general, the nature of the carrier will depend on the mode of administration being employed. For instance, parenteral formulations usually comprise injectable fluids that include pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, sesame oil, glycerol, ethanol, combinations thereof, or the like, as vehicle. The carrier and composition can be sterile, and the formulation suits the mode of administration. In addition to biological neutral carriers, pharmaceutical compositions to be administrated can contain minor amounts of non toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents and the like, for example sodium acetate or sorbitan monolaurate. The composition can be a liquid solution, suspension, emulsion.
The present invention relates to the use of at least one human Tr1 cell population directed against a human HSP for the preparation of a medicament or a pharmaceutical composition as described here above for treating an inflammatory autoimmune condition.
One object of the invention is a medicament or a pharmaceutical composition as described here above for treating an inflammatory autoimmune condition or for use in treating an inflammatory autoimmune condition.
An inflammatory autoimmune condition includes but is not limited to: autoimmune (Hashimoto's) thyroiditis, hyperthyroidism (Grave's disease), autoimmune adrenal insufficiency (Addison's disease), autoimmune oophoritis, autoimmune orchitis, autoimmune hepatitis, autoimmune hemolytic anemia, paroxysmal cold hemoglobinuria, autoimmune thrombocytopenia, autoimmune neutropenia, pernicius anemia, pure red cell anemia, autoimmune coagulopathies, myasthenia gravis, autoimmune polyneuritis, multiple sclerosis, pemphigus and other bullous diseases, rheumatic carditis, Goodpasture's syndrome, postcardiotomy syndrome, systemic lupus erythematosus, Sjorgen's syndrome, polymyositis, dermatomyositis, scleroderma, inflammatory bowel diseases: Crohn's disease, ulcerative colitis; chronic obstructive pulmonary diseases, chronic inflammatory diseases, coeliac disease, vasculitis, Wegener's disease, Churg-Strauss syndrome, primary biliary cirrhosis, primary sclerosing cholangitis, cardiovascular disease, rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, juvenile idiopathic arthritis, polydermatomyositis, septic shock, host versus graft disease, graft versus host disease, asthma, rhinitis, psoriasis, cachexia associated with cancer, eczema, vitiligo, Reiter's syndrome, Kawasaki's disease, idiopathic thrombocytopenic purpura, Guillain-Barré syndrome, antiphospholipid antibody syndrome (APS), atherosclerosis, narcolepsy.
In a preferred embodiment, the present invention relates to the use of at least one human Tr1 cell population directed against a human HSP for the preparation of a medicament or a pharmaceutical composition as described here above for treating an inflammatory autoimmune condition, which is not diabetes.
Preferably, said inflammatory autoimmune disease is selected in the group comprising intestinal inflammatory condition such as Crohn's disease and ulcerative colitis; arthritis condition such as rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis and juvenile idiopathic arthritis; multiple sclerosis, Wegener's disease, primary biliary cirrhosis, primary sclerosing cholangitis, asthma, transplant rejection (host versus graft disease), graft versus host disease.
More preferably, said inflammatory autoimmune disease is selected in the group of rheumatoid arthritis, Crohn's disease, ulcerative colitis, asthma and transplant rejection or graft versus host disease.
In a preferred embodiment, the present invention relates to the use of a composition as described here above for the preparation of a medicament or a pharmaceutical composition for treating arthritis such as rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis and juvenile idiopathic arthritis.
In another preferred embodiment, the present invention relates to the use of a composition as described here above for the preparation of a medicament or a pharmaceutical composition for treating multiple sclerosis.
In another preferred embodiment, the present invention relates to the use of a composition as described here above for the preparation of a medicament or a pharmaceutical composition for treating intestinal inflammatory condition, preferably for treating Crohn's disease and ulcerative colitis.
In another preferred embodiment, the present invention relates to the use of a composition as described here above for the preparation of a medicament or a pharmaceutical composition for treating vasculitis such as Wegener's disease and atherosclerosis.
In another preferred embodiment, the present invention relates to the use of a composition as described here above for the preparation of a medicament or a pharmaceutical composition for treating autoimmune inflammation of the biliary duct or the liver such as primary biliary cirrhosis and primary sclerosing cholangitis.
In another preferred embodiment, the present invention relates to the use of a composition as described here above for the preparation of a medicament or a pharmaceutical composition for treating transplantation related disorders such as host versus graft disease and graft versus host disease.
In another preferred embodiment, the present invention relates to the use of a composition as described here above for the preparation of a medicament or a pharmaceutical composition for treating asthma.
An object of the present invention is also a method for treating an inflammatory autoimmune condition in a subject in need thereof, comprising administering to said subject an effective amount of the medicament or pharmaceutical composition as described here above.
The composition may be formulated for parenteral, intramuscular, intravenous, intraperitoneal, injection, intranasal inhalation, lung inhalation, intradermal, intra-articular, intrathecal injection.
Preferably, the medicament or pharmaceutical composition of the invention may be administrated by intramuscular, intraperitoneal or intravenous injection, or by direct injection into the lymph nodes of the patient, more preferably by intravenous injection.
The amount of Tr1 cells directed to a human HSP effective in the treatment of an inflammatory autoimmune condition will depend on the nature of the inflammation, and can be determined by standard clinical techniques. The precise dose to be employed in the formulation will also depend on the route of administration, and the seriousness of the disease or disorder, and should be decided according to the judgment of the practitioner and each individual's circumstances. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.
In one embodiment of the present invention, 104/kg to 109/kg cells are administrated to the subject. Preferably 105/kg to 107/kg cells and more preferably about 106/kg cells are administrated to the subject.
In one embodiment of the invention, the subject is administrated with the medicament at the time when flare-up are demonstrated by a decline in the clinical status of the subject.
In one embodiment of the invention, the subject is administrated once with the medicament or the pharmaceutical composition of the present invention.
In a second embodiment of the invention, the subject is administrated once a month with the medicament or the pharmaceutical composition of the present invention.
In a third embodiment of the invention, the subject is administrated once a quarter with the medicament or the pharmaceutical composition of the present invention.
In a fourth embodiment of the invention, the subject is administrated once to twice a year with the medicament or the pharmaceutical composition of the present invention.
In another embodiment of the present invention, the medicament or pharmaceutical composition to be administered to a subject in need thereof comprises human Tr1 cells autologous to the cells of said subject.
This means that Tr1 cells will be administrated to the subject they come from or that precursors used for the production of Tr1 cells come from the subject the Tr1 cells will be administrated to.
The present invention relates also to a process for treating an inflammatory autoimmune condition in a subject in need thereof, said process comprising the steps of:
Preferably, cloning and expansion of Tr1 clones directed to a human HSP is carried out with the following method:
Examples of factors which interact with the above mentioned cell surface proteins include:
In another embodiment of the invention, the process for treating an inflammatory autoimmune condition comprises additional steps such as combined administration of one or more therapeutic agents such as described here after and/or combined administration of an effective amount of a medicament or pharmaceutical composition comprising at least one human Tr1 cell population directed against an antigen known to be involved in the inflammatory autoimmune condition as described here after.
In another embodiment of the present invention, the method for treating an inflammatory autoimmune condition in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with one or more therapeutic agents used for treating an inflammatory autoimmune condition.
The present invention relates to the use of the pharmaceutical composition or medicament of the invention for treating an inflammatory autoimmune condition, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with one or more therapeutic agents used for treating an inflammatory autoimmune condition.
Examples of therapeutic agents commonly used for treating an arthritic inflammatory condition are the following:
In a preferred embodiment of the present invention, the method for treating an arthritic inflammatory condition in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with one or more therapeutic agents selected in the group of corticoids, TNF blocking agents, anti-interleukins, anti-B lymphocytes, anti-costimulatory molecules, tolerogenic agents, anti-complement proteins, inhibitors of T cell signalling molecules, inhibitors of cell migration, methotrexate, leflunomide, sulfasalazine, hydroxychloroquine, azathioprine, methotrexate, cyclosporine, minocycline, D-penicillamine.
The present invention relates to the use of the pharmaceutical composition or medicament of the invention for treating an inflammatory arthritic condition in a subject in need thereof, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with one or more therapeutic agents selected in the group of corticoids, TNF blocking agents, anti-interleukins, anti-B lymphocytes, anti-costimulatory molecules, tolerogenic agents, anti-complement proteins, inhibitors of T cell signalling molecules, inhibitors of cell migration, methotrexate, leflunomide, sulfasalazine, hydroxychloroquine, azathioprine, methotrexate, cyclosporine, minocycline, D-penicillamine.
Examples of therapeutic agents commonly used for treating multiple sclerosis are the following:
Examples of combination therapies currently used are:
In a preferred embodiment of the present invention, the method for treating multiple sclerosis in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with one or more therapeutic agents in the group of interferon-beta, glatiramer acetate, mitoxantrone, cyclophosphamide, methotrexate, aziathropine or natalizumab.
The present invention relates to the use of the pharmaceutical composition or medicament of the invention for treating multiple sclerosis, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with one or more therapeutic agents in the group of interferon-beta, glatiramer acetate, mitoxantrone, cyclophosphamide, methotrexate, aziathropine or natalizumab.
Examples of therapeutic agents commonly used for treating an intestinal inflammatory condition are the following:
In a preferred embodiment of the present invention, the method for treating an intestinal inflammatory condition in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with one or more therapeutic agents selected in the group of anti-TNF or TNF blocking agents, natalizumab, anti-ILL anti-IL-6, anti-IL-12, anti-IL-17 and anti-IL-23; IL-1 receptor antagonist analogs (anakinra); 5 aminosalicyclic acid and analogs such as mesalazine, sulfazaline, olsalazine, balsalazide; corticoids such as prednisone, budesonide, hydrocortisone, prednisolone, methylprednisolone, betamethasone, bedomethasone, tixocortol.
The present invention relates to the use of the pharmaceutical composition or medicament of the invention for treating an intestinal inflammatory condition in a subject in need thereof, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with one or more therapeutic agents selected in the group of anti-TNF or TNF blocking agents, natalizumab, anti-ILE anti-IL-6, anti-IL-12, anti-IL-17 and anti-IL-23; IL-1 receptor antagonist analogs (anakinra); 5 aminosalicyclic acid and analogs such as mesalazine, sulfazaline, olsalazine, balsalazide; corticoids such as prednisone, budesonide, hydrocortisone, prednisolone, methylprednisolone, betamethasone, bedomethasone, tixocortol.
Therapeutic agents commonly used for treating vasculitis such as atherosclerosis and Wegener′ disease are statins, aspirin, blood coagulants such as heparin, coumadin for atherosclerosis and corticoids, aziathioprine, methothrexate, cyclophosphamide, anti-B lymphocytes antibodies (Rituximab), anti-TNF antibodies (Etanercept, Remicade) and anti-thymocyte globulin for Wegener's disease.
In a preferred embodiment of the present invention, the method for treating vasculitis such as atherosclerosis and Wegener's disease in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with statins, aspirin, blood coagulants such as heparin, coumadin for atherosclerosis and corticoids, aziathioprine, methothrexate, cyclophosphamide, anti-B lymphocytes antibodies (Rituximab) TNF blocking agents (Etanercept, Remicade) and anti-thymocyte globulin for Wegener's disease.
The present invention relates to the use of the pharmaceutical composition or medicament of the invention for treating vasculitis such as atherosclerosis and Wegener's disease in a subject in need thereof, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with statins, aspirin, blood coagulants such as heparin, coumadin for atherosclerosis and corticoids, aziathioprine, methothrexate, cyclophosphamide, anti-B lymphocytes antibodies (Rituximab) and TNF blocking agents (Etanercept, Remicade) and anti-thymocyte globulin for Wegener's disease.
Therapeutic agents commonly used for treating autoimmune inflammation of the biliary duct or the liver such as primary biliary cirrhosis and primary sclerosing cholangitis is ursodeoxycholic acid (Ursodiol).
In a preferred embodiment of the present invention, the method for treating autoimmune inflammation of the biliary duct or the liver such as primary biliary cirrhosis and primary sclerosing cholangitis in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with ursodeoxycholic acid (Ursodiol).
The present invention relates to the use of the pharmaceutical composition or medicament of the invention for treating autoimmune inflammation of the biliary duct or the liver such as primary biliary cirrhosis and primary sclerosing cholangitis in a subject in need thereof, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with ursodeoxycholic acid (Ursodiol).
Therapeutic agents commonly used for treating inflammation related to transplant rejection (host versus graft disease or graft versus host disease) are calcineurins inhibitors (Cyclosporin, Tacrolimus), mTOR inhibitors (Sirolimus, Everolimus), anti-proliferative agents (Azathioprine, Mycophenolic acid), monoclonal antibodies directed to CD25 (Dacluzimab, Basiliximab) or anti-thymocyte and anti-lymphocyte globulin preparations.
In a preferred embodiment of the present invention, the method for treating inflammation related to transplant rejection (host versus graft disease or graft versus host disease) in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with calcineurins inhibitors (Cyclosporin, Tacrolimus), mTOR inhibitors (Sirolimus, Everolimus), anti-proliferative agents (Azathioprine, Methothrexate, Mycophenolic acid), monoclonal antibodies directed to CD25 (Dacluzimab, Basiliximab) or anti-thymocyte and anti-lymphocyte globulin preparations.
The present invention relates to the use of the pharmaceutical composition or medicament of the invention for treating inflammation related to transplant rejection (host versus graft disease or graft versus host disease) in a subject in need thereof, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with calcineurins inhibitors (Cyclosporin, Tacrolimus), mTOR inhibitors (Sirolimus, Everolimus), anti-proliferative agents (Azathioprine, Mycophenolic acid), monoclonal antibodies directed to CD25 (Dacluzimab, Basiliximab) or anti-thymocyte and anti-lymphocyte globulin preparations.
Therapeutic agents commonly used for treating asthma are short-acting, selective beta2-adrenoceptor agonists, such as salbutamol (albuterol USAN), levalbuterol, terbutaline and bitolterol and other adrenergic agonists such as inhaled epinephrine and ephedrine tablets, anticholinergic medications such as ipratropium bromide, inhaled glucocorticoids.
In a preferred embodiment of the present invention, the method for treating asthma in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with short-acting, selective beta2-adrenoceptor agonists such as salbutamol (albuterol USAN), levalbuterol, terbutaline and bitolterol and other adrenergic agonists such as inhaled epinephrine and ephedrine tablets, anticholinergic medications, such as ipratropium bromide, inhaled glucocorticoids.
The present invention relates to the use of the pharmaceutical composition or medicament of the invention for treating asthma in a subject in need thereof, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with short-acting, selective beta2-adrenoceptor agonists such as salbutamol (albuterol USAN), levalbuterol, terbutaline and bitolterol and other adrenergic agonists, such as inhaled epinephrine and ephedrine tablets, anticholinergic medications such as ipratropium bromide, inhaled glucocorticoids.
In another embodiment of the present invention, the method for treating an inflammatory autoimmune condition in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with one or more medicaments or pharmaceutical compositions comprising at least one human Tr1 cell population directed against an antigen known to be involved in the inflammatory autoimmune condition.
The present invention relates to the use of the pharmaceutical composition or medicament of the invention, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with one or more medicaments or pharmaceutical compositions comprising at least one human Tr1 cell population directed against an antigen known to be involved in the inflammatory autoimmune condition.
Examples of medicament or pharmaceutical composition that can be used for treating an arthritic condition, such as rheumatoid arthritis, ankylosing spondylitis, juvenile idiopathic arthritis and psoriatic arthritis, are the medicament or pharmaceutical composition comprising at least one Tr1 cell population or clone directed against a joint-associated antigen. Said joint-associated antigen is for example citrulline-substituted cyclic and linear filaggrin peptides, collagen type II peptides, human cartilage glycoprotein 39 (HCgp39) peptides, heterogenous nuclear ribonucleoprotein (hnRNP) A2 peptides, hnRNP B1, hnRNP D, Ro60/52, BiP, keratin, vimentin, fibrinogen, cardiolipin, collagen type I, III, IV and V peptides, annexin V, Glucose 6 phosphate isomerase (GPI), acetyl-calpastatin, pyruvate deshydrogenase (PDH), aldolase, topoisomerase I, snRNP, PARP, Scl-70, Scl-100, phospholipid antigen including anionic phosphatidylserine, neutrally charged phosphatidylethanolamine and phosphatidylcholine, matrix metalloproteinase, fibrillin, aggreccan, and fragments, variants and mixtures thereof.
In a preferred embodiment, said human Tr1 cell population or clone is directed against a joint-associated antigen selected among type II collagen, HCgp39, fragments and variants thereof.
In a preferred embodiment of the invention, the method for treating an arthritic condition in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with an effective amount of a medicament or a pharmaceutical composition comprising at least one Tr1 cell population or clone directed against a joint-associated antigen, preferably against type II collagen, HCgp39, fragments, variants and mixtures thereof.
The present invention relates to the use of the pharmaceutical composition or medicament of the invention for treating an arthritic condition in a subject in need thereof, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with an effective amount of a medicament or a pharmaceutical composition comprising at least one Tr1 cell population or clone directed against a joint-associated antigen, preferably against type II collagen, HCgp39, fragments, variants and mixtures thereof.
Examples of medicament or pharmaceutical composition that can be used for treating a multiple sclerosis condition are the medicament or pharmaceutical composition comprising at least one Tr1 cell population or clone directed against a multiple sclerosis-associated antigen. Said multiple sclerosis-associated antigen is for example myelin basic protein, myelin associated glycoprotein, myelin oligodendrocyte protein, proteolipid protein, oligodendrocyte myelin oligoprotein, myelin associated oligodendrocyte basic protein, oligodendrocyte specific protein, oligodendrocyte specific proteins, NOGO A, glycoprotein Po, peripheral myelin protein 22, 2′3′-cyclic nucleotide 3′-phosphodiesterase, and fragments, variants and mixtures thereof.
In a preferred embodiment, said human Tr1 cell population or clone is directed against a multiple sclerosis-associated antigen selected among myelin basic protein (MBP), proteolipid protein (PLP) and myelin oligodendrocyte protein (MOG) and fragments, variants and mixtures thereof.
In a preferred embodiment of the invention, the method for treating a multiple sclerosis condition in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with an effective amount of a medicament or a pharmaceutical composition comprising at least one Tr1 cell population or clone directed against a multiple sclerosis-associated antigen, preferably against myelin basic protein (MBP), proteolipid protein (PLP) and myelin oligodendrocyte protein (MOG) and fragments, variants and mixtures thereof.
The present invention relates to the use of the pharmaceutical composition or medicament of the invention for treating a multiple sclerosis condition in a subject in need thereof, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with an effective amount of a medicament or a pharmaceutical composition comprising at least one Tr1 cell population or clone directed against a multiple sclerosis-associated antigen, preferably against myelin basic protein (MBP), proteolipid protein (PLP) and myelin oligodendrocyte protein (MOG) and fragments, variants and mixtures thereof.
Examples of medicament or pharmaceutical composition that can be used for treating an intestinal inflammatory condition, such as Crohn's disease and ulcerative colitis, are the medicament or pharmaceutical composition comprising at least one Tr1 cell population or clone directed against a food antigen from common human diet. Said food antigen from common human diet is for example bovine antigens, Ca-binding SlOO, alpha-lactalbumin, lactoglobulins, bovine serum albumin, immunoglobulin or caseins, atlantic salmon antigens, chicken antigens, ovalbumin, Ag22, conalbumin, lysozyme or chicken serum albumin, shrimp antigens, wheat antigens, celery antigens, carrot antigens, apple antigens, apple lipid transfer protein, apple profilin, pear antigens, isoflavone reductase, avocado antigens, apricot antigens, peach antigens, soybean antigens, peanuts, and fragments, variants and mixtures thereof.
In a preferred embodiment, said human Tr1 cell population or clone is directed against a food antigen from common human diet selected among ovalbumin, casein, beta-lactoglobulin, soya protein, gliadin, peanuts, and fragments, variants and mixtures thereof.
In a preferred embodiment of the invention, the method for treating an intestinal inflammatory condition in a subject in need thereof comprises the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention in combination with an effective amount of a medicament or a pharmaceutical composition comprising at least one Tr1 cell population or clone directed against a food antigen from common human diet, preferably against ovalbumin, casein, beta-lactoglobulin, soya protein, gliadin, peanuts, and fragments, variants and mixtures thereof. The present invention relates to the use of the pharmaceutical composition or medicament of the invention for treating an intestinal inflammatory condition in a subject in need thereof, wherein the administration to said subject of an effective amount of the medicament or the pharmaceutical composition of the invention is in combination with an effective amount of a medicament or a pharmaceutical composition comprising at least one Tr1 cell population or clone directed against a food antigen from common human diet, preferably against ovalbumin, casein, beta-lactoglobulin, soya protein, gliadin, peanuts, and fragments, variants and mixtures thereof.
In the following description, all experiments for which no detailed protocol is given are performed according to standard protocol.
The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.
Blood samples from healthy patients were collected and white blood cells were separated using gradient density centrifugation. Cells were then cultured in the presence of human HSP60 in order to induce the specific proliferation of Tr1 cells directed against this antigen. After 13 days of culture, cell populations were cloned by limiting dilution method. Clones were then assessed for their specificity to human HSP60 and for characteristic Tr1 cytokine production profile.
For the determination of antigen specificity, sandwich ELISAs were performed on 48 hours supernatants of Tr1 cell clones stimulated in the presence of antigen presenting cells (4.105) and in the presence or absence of the specific antigen (HSP60). For the determination of the cytokine production profile, HSP60 Tr1 cell clones were stimulated with anti-CD3+anti-CD28 monoclonal antibodies and the supernatants were harvested after 48 hours. ELISAs were performed using anti-IL-4 (11B11), anti-IL-10 (2A5), anti-IFN-γ (XGM1.2), biotin anti-IL-4 (24G2), anti-IL-10 (SXC1), anti-IFN-γ (R4-6A2) (Pharmingen Becton Dickinson).
For suppression studies, autologous CD4 positive T lymphocytes were co-cultured with or without supernatants from activated HSP-60 specific Tr1 clones. Cultures were stimulated with anti-CD3+anti-CD28 monoclonal antibodies and Interleukin-2. After 5 days total cell proliferation was assessed using the WST-1 proliferation kit from Roche.
To further determine the cytokine secretion profile of these Tr1 cell populations specific for HSP60, cells were stimulated in the presence of anti-CD3+anti-CD28 monoclonal antibodies. ELISAs were performed on 48 h supernatants to measure IL-4, IL-10 and IFNγ production.
Suppressive activity of these HSP60 Tr1 populations was then assessed. Autologous CD4 positive T lymphocytes were co-cultured with or without supernatants from activated HSP-60 specific Tr1 clones. After 5 days of stimulation, cell proliferation was measured.
| Number | Date | Country | Kind |
|---|---|---|---|
| 08305134.2 | Apr 2008 | EP | regional |
| Number | Date | Country | |
|---|---|---|---|
| 61048298 | Apr 2008 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 12989878 | Oct 2010 | US |
| Child | 14796284 | US |
