Compositions with reduced dimer formation

Abstract

The present invention relates to the use of a non-reducing carbohydrate or carbohydrate derivative and at least one agent which inhibits dimer formation in a freeze-dried composition comprising at least one peptide that contains a free cysteine residue, to provide a freeze-dried composition with improved long-term storage stability.

1 - 19 . (canceled) 
     
     
         20 . A method of producing a stable freeze-dried composition comprising at least one peptide that is 8 to 30 amino acids in length and that comprises at least one free cysteine residue, said method comprising:
 (a) preparing a composition comprising in solution: (i) at least one non-reducing carbohydrate selected from trehalose, sucrose, raffinose, stachyose or melezitose; (ii) thioglycerol; and (iii) at least one said peptide; and   (b) freeze-drying the composition resulting from step (a).   
     
     
         21 . A method according to  claim 20 , wherein:
 (a) the non-reducing carbohydrate is trehalose; and/or   (b) the thioglycerol is present at a molar concentration which is between 60 and 80 fold, or between 30 and 40 fold, greater than the molar concentration of the peptide components of the composition comprising at least one free cysteine residue.   
     
     
         22 . A method according to  claim 20 , wherein the at least one peptide comprises a region comprising at least one MHC-binding T cell epitope derived from an allergen or allo-antigen, and optionally the epitope derives from:
 (a) an allergen selected from: a plant allergen (particularly a grass allergen), animal dander allergens, a mold or fungal allergen, a dust allergen, an antibiotic or other drug, a stinging insect venom, an environmental allergen or a food allergen;   (b) an antigen selected from the major antigens associated with Acute disseminated encephalomyelitis (ADEM); Addison's disease; Ankylosing spondylitis; Antiphospholipid antibody syndrome (APS); Aplastic anemia; Autoimmune hepatitis; Autoimmune Oophoritis; Coeliac disease; Crohn's disease; Diabetes mellitus type 1; Gestational pemphigoid; Goodpasture's syndrome; Graves' disease; Guillain-Barré syndrome (GBS); Hashimoto's disease; Idiopathic thrombocytopenic purpura; Kawasaki's Disease; Lupus erythematosus; Multiple sclerosis; Myasthenia gravis; Narcolepsy, Opsoclonus myoclonus syndrome (OMS); Optic neuritis; Ord's thyroiditis; Pemphigus; Pernicious anaemia; Polyarthritis in dogs; Primary biliary cirrhosis; Rheumatoid arthritis; Reiter's syndrome; Sjögren's syndrome; Takayasu's arteritis; Temporal arteritis (also known as “giant cell arteritis”); Warm autoimmune hemolytic anemia; or Wegener's granulomatosis; or   (c) cat dander protein Fel d1; House dust mite proteins Der p 1, Der p 2, Der p 7, Der p 3 to 15, Der p 18, 20, 21 and 23, Der f 1, 2, 7, 10, 11 to 18 and 20 to 22; Ragweed protein amb a 1, 2, 3, 5, 6, 7, 8, 9 and isoforms thereof including amb a 1.1, a 1.2, a1.3 or a1.4; Rye grass proteins lol p 1 and lol p5; Timothy grass proteins phl p1 and phl p5; Bermuda grass protein Cyn d 5;  Alternaria alternata  proteins Alt a1, Alt a 2, Alt a 3 to Alt a 10, Alt a 12, Alt a 13 and Enolase (Alt a 6), Cla h 1, 2, 5 to 10, 12, GST, HCh1, HSP70, NTF2, TCTP; Birch protein Bet v 1, 2, 3, 4, 6, 7, 8 and P14; German Cockroach proteins Bla g 1, Bla g 2, Bla g 3, Bla g 4, Bla g 5 and Bla g 6, 7, 8, 9, GSTD1, Trypsin; Mugwort protein Art v 1; Russian thistle protein Sal k 1, Sal k 2 and Sal k 8; peanut Ara h1, Ara h2, Ara h3, Ara h4, Ara h5, Ara h6, plant profilins or lipid transfer proteins or a human leukocyte antigen.   
     
     
         23 . A method according to  claim 20 , wherein:
 (a) the at least one peptide comprises or consists of the sequence of any of SEQ ID NOS. 1 to 58;   (b) the composition comprises the peptides of SEQ ID NOS. 1 to 4; or   (c) the composition comprises the peptides of SEQ ID NOS. 1 to 4 and further comprises the peptides of SEQ ID NOS. 84 to 86, and optionally no further peptides.   
     
     
         24 . A stable, freeze-dried composition produced according to the method of  claim 20 . 
     
     
         25 . A stable, freeze-dried composition comprising at least one peptide that is 8 to 30 amino acids in length and that comprises at least one cysteine residue, thioglycerol and at least one non-reducing carbohydrate selected from trehalose, sucrose, raffinose, stachyose or melezitose. 
     
     
         26 . A composition according to  claim 25 , wherein:
 (a) the non-reducing carbohydrate is trehalose; and/or   (b) the thioglycerol is present at a molar concentration which is between 60 and 80 fold, or between 30 and 40 fold, greater than the molar concentration of the peptide components of the composition comprising at least one free cysteine residue.   
     
     
         27 . A composition according to  claim 25 , wherein the at least one peptide comprises a region comprising at least one MHC-binding T cell epitope derived from an allergen or allo-antigen, and optionally the epitope derives from:
 (a) an allergen selected from: a plant allergen (particularly a grass allergen), animal dander allergens, a mold or fungal allergen, a dust allergen, an antibiotic or other drug, a stinging insect venom, an environmental allergen or a food allergen;   (b) an antigen selected from the major antigens associated with Acute disseminated encephalomyelitis (ADEM); Addison's disease; Ankylosing spondylitis; Antiphospholipid antibody syndrome (APS); Aplastic anemia; Autoimmune hepatitis; Autoimmune Oophoritis; Coeliac disease; Crohn's disease; Diabetes mellitus type 1; Gestational pemphigoid; Goodpasture's syndrome; Graves' disease; Guillain-Barré syndrome (GBS); Hashimoto's disease; Idiopathic thrombocytopenic purpura; Kawasaki's Disease; Lupus erythematosus; Multiple sclerosis; Myasthenia gravis; Narcolepsy, Opsoclonus myoclonus syndrome (OMS); Optic neuritis; Ord's thyroiditis; Pemphigus; Pernicious anaemia; Polyarthritis in dogs; Primary biliary cirrhosis; Rheumatoid arthritis; Reiter's syndrome; Sjögren's syndrome; Takayasu's arteritis; Temporal arteritis (also known as “giant cell arteritis”); Warm autoimmune hemolytic anemia; or Wegener's granulomatosis; or   (c) cat dander protein Fel d1; House dust mite proteins Der p 1, Der p 2, Der p 7, Der p 3 to 15, Der p 18, 20, 21 and 23, Der f 1, 2, 7, 10, 11 to 18 and 20 to 22; Ragweed protein amb a 1, 2, 3, 5, 6, 7, 8, 9 and isoforms thereof including amb a 1.1, a 1.2, a1.3 or a1.4; Rye grass proteins lol p 1 and lol p5; Timothy grass proteins phl p1 and phl p5; Bermuda grass protein Cyn d 5;  Alternaria alternata  proteins Alt a1, Alt a 2, Alt a 3 to Alt a 10, Alt a 12, Alt a 13 and Enolase (Alt a 6), Cla h 1, 2, 5 to 10, 12, GST, HCh1, HSP70, NTF2, TCTP; Birch protein Bet v 1, 2, 3, 4, 6, 7, 8 and P14; German Cockroach proteins Bla g 1, Bla g 2, Bla g 3, Bla g 4, Bla g 5 and Bla g 6, 7, 8, 9, GSTD1, Trypsin; Mugwort protein Art v 1; Russian thistle protein Sal k 1, Sal k 2 and Sal k 8; peanut Ara h1, Ara h2, Ara h3, Ara h4, Ara h5, Ara h6, plant profilins or lipid transfer proteins or a human leukocyte antigen.   
     
     
         28 . A composition according to  claim 25 , wherein:
 (a) the at least one peptide comprises or consists of the sequence of any of SEQ ID NOS. 1 to 58;   (b) the composition comprises the peptides of SEQ ID NOS. 1 to 4; or   (c) the composition comprises the peptides of SEQ ID NOS. 1 to 4 and further comprises the peptides of SEQ ID NOS. 84 to 86, and optionally no further peptides.   
     
     
         29 . A method of reconstituting a stable freeze-dried composition which comprises at least one peptide that is 8 to 30 amino acids in length and that comprises at least one cysteine residue, thioglycerol and at least one non-reducing carbohydrate selected from trehalose, sucrose, raffinose, stachyose or melezitose, wherein said method comprises reconstituting the said freeze-dried composition in solution. 
     
     
         30 . A solution reconstituted by a method as defined in  claim 29 .