Therapeutically active aldesleukin highly stable in liquid pharmaceutical compositions

Abstract

The disclosure relates to a liquid pharmaceutical composition of aldesleukin/SDS aggregates and its use in the treatment of auto-immune disease, inflammatory disorders, gene therapies and cancer. A method for preparing said composition is also described.

What is claimed,: 
     
         1 . A pharmaceutical composition comprising IL-2 in a range from about 0.001 mg to 3 mg/mL, sodium dodecyl sulfate (SDS) from about 0.0001 mg/ml to about 10 mg/mL, a non-ionic osmolytes(s), or combinations thereof. 
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein the IL-2 comprises SEQ ID NO: 1 or variants thereof. 
     
     
         3 . The pharmaceutical composition of  claim 1 , wherein the osmolytes comprise mannitol, sorbitol, xylitol or a pH 7-8 phosphate buffer. 
     
     
         4 . A method of inducing proliferation of regulatory T cells in vivo, comprising administering to a subject in need thereof, a therapeutically effective amount of aldesleukin wherein the regulatory T cells increase as compared to a control, inducing proliferation of regulatory T cells in vivo. 
     
     
         5 . The method of  claim 4 , wherein the aldesleukin is comprised in a pharmaceutical composition. 
     
     
         6 . The method of  claim 4 , wherein the aldesleukin comprises SEQ ID NO: 1 or variants thereof 
     
     
         7 . The method of  claim 1 , wherein the aldesleukin remains stable in the pharmaceutical liquid composition for at least a year as measured by an in vitro biological activity assay. 
     
     
         8 . The method of  claim 1 , wherein the aldesleukin comprises a stable aldesleukin/SDS aggregate distribution in the liquid composition at least for a year in a range of about 4-18 nm with a peak at about 8 nm as measured by dynamic light scattering. 
     
     
         9 . The method of  claim 1 , wherein the aldesleukin has a stable therapeutic activity, at least for a year in the liquid composition, as measured in an animal model of human cancer. 
     
     
         10 . A method of treating an autoimmune disease or disorder in a subject in need thereof, comprising: administering to a subject in need thereof, a pharmaceutical composition comprising a therapeutically effective amount of aldesleukin wherein the regulatory T cells increase as compared to a control, inducing proliferation of regulatory T cells in vivo. 
     
     
         11 . The method of  claim 10 , wherein the aldesleukin remains stable in the pharmaceutical liquid composition for at least a year as measured by an in vitro biological activity assay. 
     
     
         12 . The method of  claim 6 , wherein the aldesleukin comprises a stable aldesleukin/SDS aggregate distribution in the liquid composition at least for a year in a range of about 4-18 nm with a peak at about 8 nm as measured by dynamic light scattering. 
     
     
         13 . The method of  claim 6 , wherein the aldesleukin has a stable therapeutic activity, at least for a year in the liquid composition, as measured in an animal model of human cancer. 
     
     
         14 . A method of regulating an immune response associated with a disease or disorder in a subject in need thereof, comprising: administering to a subject in need thereof, a pharmaceutical composition comprising a therapeutically effective amount of aldesleukin wherein the regulatory T cells increase as compared to a control, inducing proliferation of regulatory T cells in vivo. 
     
     
         15 . The method of  claim 10 , wherein the aldesleukin remains stable in the pharmaceutical liquid composition for at least a year as measured by an in vitro biological activity assay. 
     
     
         16 . The method of  claim 10 , wherein the aldesleukin comprises a stable aldesleukin/SDS aggregate distribution in the liquid composition at least for a year in a range of about 4-18 nm with a peak at about 8 nm as measured by dynamic light scattering. 
     
     
         17 . The method of  claim 10 , wherein the aldesleukin has a stable therapeutic activity, at least for a year in the liquid composition, as measured in an animal model of human cancer. 
     
     
         18 . The method of  claim 10 , wherein the disease or disorder comprises: an autoimmune disease, cancer, inflammation, a viral infection, a bacterial infection, a neurodegenerative disorder or combinations thereof. 
     
     
         19 . A method of producing aldesleukin suitable for the preparation of a composition comprising aldesleukin in a pharmaceutical aqueous vehicle, the method comprising:
 fermenting a bacterial cell transfected with an expression vector encoding an aldesleukin gene;   disrupting the bacterial cell;   collecting inclusion bodies containing aldesleukin;   subjecting the inclusion bodies to dissolution by a detergent followed by oxidation and a first chromatography;   diluting with an organic nitrile and subjecting to a second chromatography;   subjecting the aldesleukin to diafiltration and sterilization.   
     
     
         20 . The method of  claim 19 , wherein the bacterial cell is an  Escherichia coli  bacterial cell. 
     
     
         21 . The method of  claim 19 , wherein the detergent is sodium dodecyl sulfate (SDS). 
     
     
         22 . The method of  claim 19 , wherein the oxidation is conducted with an oxidizing compound. 
     
     
         23 . The method of  claim 19 , wherein the first chromatography is a ceramic hydroxyapatite chromatography. 
     
     
         24 . The method of  claim 19 , wherein the second chromatography is a high-performance liquid chromatography (HPLC). 
     
     
         25 . The method of  claim 24 , wherein the HPLC is C4 column HPLC chromatography. 
     
     
         26 . The method of  claim 1 , wherein the aldesleukin remains stable in the pharmaceutical liquid composition for at least a year as measured by an in vitro biological activity assay. 
     
     
         27 . The method of  claim 19 , wherein the aldesleukin comprises a stable aldesleukin/SDS aggregate distribution in the liquid composition at least for a year in a range of about 4-18 nm with a peak at about 8 nm as measured by dynamic light scattering. 
     
     
         28 . The method of  claim 19 , wherein the aldesleukin has a stable therapeutic activity, at least for a year in the liquid composition, as measured in an animal model of human cancer. 
     
     
         29 . The method of  claim 19 , wherein the aldesleukin is used as a drug for medical treatments involving cells expressing IL-2 receptors in their membranes, comprising NK cells, T cells, dendritic cells, nonlymphoid cells, cell lines, transformed cells or combinations thereof. 
     
     
         30 . A pharmaceutical composition comprising aldesleukin in which the aldesleukin: a) remains stable in the pharmaceutical liquid composition for at least a year as measured by the in vitro biological activity assay, b) have a stable aldesleukin/SDS aggregate distribution in the liquid composition at least for a year in the range of 4-18 nm range with a peak at about 8 nm as measure by dynamic light scattering, c) have a stable therapeutic activity, at least for a year in the liquid composition, as measure in an animal model of human cancer and d) is adequate to be used as a drug for treatments involving cells expressing IL-2 receptors in their membranes. 
     
     
         31 . A method of restoring T-cell mediated immune responses in a subject in need thereof comprising administering a therapeutically effective amount of (i) a pharmaceutical composition comprising IL-2 in a range from about 0.001 mg to 3 mg/mL, sodium dodecyl sulfate (SDS) from about 0.0001 mg/ml to about 10 mg/mL, a non-ionic osmolytes(s), or combinations thereof (b) an expression vector encoding a therapeutic protein; (ii) one or more mobilization factors, thereby restoring T-cell mediated immune responses in the subject in need thereof. 
     
     
         32 . The method of  claim 31 , wherein the IL-2 comprises SEQ ID NO: 1 or variants thereof. 
     
     
         33 . The method of  claim 31 , wherein the osmolytes comprise mannitol, sorbitol, xylitol or a pH 7-8 phosphate buffer. 
     
     
         34 . The method of  claim 31 , wherein the subject is suffering from an immune deficiency. 
     
     
         35 . The method of  claim 34 , wherein the subject is suffering from Autoimmune Lymphoproliferative Syndrome (ALPS), APS-1 (APECED), CARD9, Chronic Granulomatous Disease (CGD), Congenital Neutropenia Syndromes, Common Variable Immunodeficiency (CVID), CTLA4 Deficiency, DOCK8 Deficiency, Glycosylation Disorders with Immunodeficiency, Hyper-Immunoglobulin E Syndromes (HIES), PI3 Kinase Disease, PLAID, Severe Combined Immunodeficiency (SCID), STAT3 Dominant-Negative Disease, WHIM Syndrome, X-Linked Agammaglobulinemia (XLA), X-Linked Lymphoproliferative Disease (XLP), Wiscott-Aldrich syndrome, DiGeorge syndrome, Ataxia-telangectasia, Chronic granulomatous disease, Transient hypogammaglobulinemia of infancy, Agammaglobulinemia, Complement deficiencies, or Selective IgA deficiency. 
     
     
         36 . A method of treating a subject suffering from an immune deficiency disease comprising administering to the subject:
 (i) an expression vector encoding SEQ ID NO: 1; and/or,   (ii) a cell comprising an expression vector encoding SEQ ID NO: 1; and/or,   (iii) a pharmaceutical composition comprising IL-2 in a range from about 0.001 mg to 3 mg/mL, sodium dodecyl sulfate (SDS) from about 0.0001 mg/ml to about 10 mg/mL, a non-ionic osmolytes(s), or combinations thereof.   
     
     
         37 . A method of treating a subject in need of immunotherapy, comprising: administering to the subject:
 (i) an expression vector encoding SEQ ID NO: 1; and/or,   (ii) a cell comprising an expression vector encoding SEQ ID NO: 1, thereby treating the subject.   
     
     
         38 . The method of  claim 37 , wherein the subject is suffering from cancer, immunodeficiency, immunosuppression, viral infections, an immune disorder or combinations thereof. 
     
     
         39 . The method of  claim 38 , wherein the immune disorder, inflammation, graft versus host disease (GVHD), transplant rejection, or an autoimmune disorder. 
     
     
         40 . The method of  claim 39 , wherein the immune disorder is multiple sclerosis, inflammatory bowel disease, rheumatoid arthritis, type I diabetes, systemic lupus erythrematosus, contact hypersensitivity, asthma or Sjogren's syndrome. 
     
     
         41 . The method of  claim 37 , further comprising administering IL-2. 
     
     
         42 . The method of  claim 37 , wherein the cells comprise: autologous cells, allogeneic cells, haplotype matched cells, haplotype mismatched cells, haplo-identical cells, xenogeneic cells, cell lines or combinations thereof. 
     
     
         43 . The method of  claim 37 , wherein the cells comprise stem cells, cord blood cells, adult stem cells, mesenchymal stem cells, mesenchymal stromal cells, induced pluripotent stem cells, autologous stem cells, bone marrow cells, hematopoietic cells, hematopoietic stem cells, somatic cells, germ line cells, differentiated cells, somatic stem cells, embryonic stem cells or combinations thereof.