Antibodies against il-1 beta

Abstract

The present invention relates to anti-IL-1 beta antibodies and in particular to monovalent high potency IL-1 beta-binding antibody fragments being highly stable. Such antibodies can be used in the treatment of inflammatory and other diseases as well as in diagnostics. Also provided are related nucleic acids, vectors, cells, and compositions.

1 . An isolated nucleic acid molecule encoding a humanized antibody against IL-1 beta comprising a variable light chain sequence having at least 90% identity to SEQ ID No: 8; and a variable heavy chain sequence having at least 90% identity to SEQ ID No: 12. 
     
     
         2 . The molecule of  claim 1 , comprising a variable heavy chain sequence (VH) as set forth in SEQ ID Nos. 9, 10, 11, 12 or 13. 
     
     
         3 . The molecule of  claim 1 , comprising a variable light chain (VL) sequence as set forth in SEQ ID Nos. 7, 8 or 24. 
     
     
         4 . The molecule of  claim 1 , wherein the antibody has the sequence as set forth in SEQ ID No.: 19. 
     
     
         5 . The molecule of  claim 1 , wherein the antibody is an antibody fragment selected from the group consisting of a Fab, a Fab′, a F(ab)′2, a scFv, a Fv fragment, a nanobody, a VHH or a minimal recognition unit. 
     
     
         6 . The molecule of  claim 1 , wherein the antibody is a scFv fragment. 
     
     
         7 . An isolated nucleic acid molecule encoding a humanized antibody against IL-1 beta comprising variable heavy chain CDR-H1, CDR-H2 and CDR-H3 sequences as set forth in SEQ ID Nos: 4, 21, and 6, respectively, or variants thereof, and wherein
 a) position 24 of the heavy chain is an alanine residue (A);   b) position 25 of the heavy chain is a phenylalanine residue (F);   c) position 44 of the heavy chain is an isoleucine residue (I);   d) position 56 of the heavy chain is a serine residue (S);   e) position 82 of the heavy chain is a lysine residue (K);   f) position 86 of the heavy chain is an arginine residue (R); and/or   g) position 105 of the heavy chain is a phenylalanine residue (F); according to AHo numbering;   and/or comprising variable light chain CDR-L1, CDR-L2 and CDR-L3 sequences as set forth in SEQ ID Nos: 1, 2; and 3, respectively, or variants thereof, and wherein   h) position 1 of the light chain is an aspartic acid residue (D);   i) position 3 of the light chain is a glutamine residue (Q);   j) position 20 of the light chain is a threonine residue (T);   k) position 99 of the light chain is a glutamic acid residue (E);   l) position 105 of the light chain is a phenylalanine residue (F);   m) position 146 of the light chain is a glutamic residue (E);   n) position 147 of the light chain is an isoleucine residue (I);   o) position 148 of the light chain is a lysine residue (K); and/or   p) position 149 of the light chain is an arginine residue (R), according to AHo numbering.   
     
     
         8 . The molecule of  claim 7 , wherein the variable heavy chain further comprises at least one of the following residues:
 (i) Serine (S) at heavy chain amino acid position 12 (according to AHo numbering);   (ii) Serine (S) or Threonine (T) at heavy chain amino acid position 103 (according to AHo numbering); and/or   (iii) Serine (S) or Threonine (T) at heavy chain amino acid position 144 (according to AHo numbering).   
     
     
         9 . The molecule of  claim 7 , wherein the variable light chain sequence comprises the sequence as set forth in SEQ ID No.: 8 and the variable heavy chain sequence comprises the sequence as set forth in SEQ ID No.: 12. 
     
     
         10 . The molecule of  claim 9 , wherein the antibody further comprises a linker sequence set forth in SEQ ID No: 14. 
     
     
         11 . The molecule of  claim 7 , wherein the antibody has the sequence as set forth in SEQ ID No.: 19. 
     
     
         12 . The molecule of  claim 7 , wherein the antibody is an antibody fragment selected from the group consisting of a Fab, a Fab′, a F(ab)′2, a scFv, a Fv fragment, a nanobody, a VHH or a minimal recognition unit. 
     
     
         13 . The molecule of  claim 7 , wherein the antibody is a scFv fragment. 
     
     
         14 . The molecule of  claim 7 , wherein the antibody is a full-length immunoglobulin. 
     
     
         15 . The molecule of  claim 7 , wherein the antibody has an IC50 with regard to human IL-1 beta of lower than 10 nM wherein the IC50 is determined by inhibiting the IL-1 beta induced release of IL-6 from human fibroblasts. 
     
     
         16 . The molecule of  claim 15 , wherein the IC50 with regard to human IL-1 beta is about 5 pM. 
     
     
         17 . The molecule of  claim 7 , wherein the antibody remains at least 80% monomeric after incubation for 1 month at room temperature. 
     
     
         18 . The molecule of  claim 7 , wherein the antibody is monovalent or multivalent. 
     
     
         19 . The molecule of  claim 7 , wherein the antibody is chemically or biologically modified. 
     
     
         20 . The molecule of  claim 19 , wherein the antibody is glycosylated, PEGylated or HESylated. 
     
     
         21 . A vector comprising a nucleic acid molecule encoding a humanized antibody against IL-1 beta comprising a variable light chain sequence having at least 90% identity to SEQ ID No: 8; and a variable heavy chain sequence having at least 90% identity to SEQ ID No: 12. 
     
     
         22 . The vector of  claim 21 , wherein the antibody comprises a variable heavy chain sequence (VH) as set forth in SEQ ID Nos. 9, 10, 11, 12 or 13. 
     
     
         23 . The vector of  claim 21 , wherein the antibody comprises a variable light chain (VL) sequence as set forth in SEQ ID Nos. 7, 8 or 24. 
     
     
         24 . The vector of  claim 21 , wherein the antibody has the sequence as set forth in SEQ ID No.: 19. 
     
     
         25 . The vector of  claim 21 , wherein the antibody is an antibody fragment selected from the group consisting of a Fab, a Fab′, a F(ab)′2, a scFv, a Fv fragment, a nanobody, a VHH or a minimal recognition unit. 
     
     
         26 . The vector of  claim 21 , wherein the antibody is a scFv fragment. 
     
     
         27 . A vector comprising an isolated nucleic acid molecule encoding a humanized antibody against IL-1 beta comprising variable heavy chain CDR-H1, CDR-H2 and CDR-H3 sequences as set forth In SEQ ID Nos: 4, 21, and 6, respectively, or variants thereof, and wherein
 a) position 24 of the heavy chain is an alanine residue (A);   b) position 25 of the heavy chain is a phenylalanine residue (F);   c) position 44 of the heavy chain is an isoleucine residue (I);   d) position 56 of the heavy chain is a serine residue (S);   e) position 82 of the heavy chain is a lysine residue (K);   f) position 86 of the heavy chain is an arginine residue (R); and/or   g) position 105 of the heavy chain is a phenylalanine residue (F); according to AHo numbering;   and/or comprising variable light chain CDR-L1, CDR-L2 and CDR-L3 sequences as set forth in SEQ ID Nos: 1, 2; and 3, respectively, or variants thereof, and wherein   h) position 1 of the light chain is an aspartic acid residue (D);   i) position 3 of the light chain is a glutamine residue (Q);   j) position 20 of the light chain is a threonine residue (T);   k) position 99 of the light chain is a glutamic acid residue (E);   l) position 105 of the light chain is a phenylalanine residue (F);   m) position 146 of the light chain is a glutamic residue (E);   n) position 147 of the light chain is an isoleucine residue (I);   o) position 148 of the light chain is a lysine residue (K); and/or   p) position 149 of the light chain is a arginine residue (R), according to AHo numbering.   
     
     
         28 . The vector of  claim 27 , wherein the variable heavy chain further comprises at least one of the following residues:
 (i) Serine (S) at heavy chain amino acid position 12 (according to AHo numbering);   (ii) Serine (S) or Threonine (T) at heavy chain amino acid position 103 (according to AHo numbering); and/or   (iii) Serine (S) or Threonine (T) at heavy chain amino acid position 144 (according to AHo numbering).   
     
     
         29 . The vector of  claim 27 , wherein the variable light chain sequence comprises the sequence as set forth in SEQ ID No.: 8 and the variable heavy chain sequence comprises the sequence as set forth in SEQ ID No.: 12. 
     
     
         30 . The vector of  claim 27 , wherein the antibody further comprises a linker sequence set forth in SEQ ID No: 14. 
     
     
         31 . The vector of  claim 27 , wherein the antibody has the sequence as set forth in SEQ ID No.: 19. 
     
     
         32 . The vector of  claim 27 , wherein the antibody is an antibody fragment selected from the group consisting of a Fab, a Fab′, a F(ab)′2, a scFv, a Fv fragment, a nanobody, a VHH or a minimal recognition unit. 
     
     
         33 . The vector of  claim 27 , wherein the antibody is a scFv fragment. 
     
     
         34 . The vector of  claim 27 , wherein the antibody is a full-length immunoglobulin. 
     
     
         35 . The vector of  claim 27 , wherein the antibody has an IC50 with regard to human IL-1 beta of lower than 10 nM wherein the IC50 is determined by inhibiting the IL-1 beta induced release of IL-6 from human fibroblasts. 
     
     
         36 . The vector of  claim 35 , wherein the IC50 with regard to human IL-1 beta is about 5 pM. 
     
     
         37 . The vector of  claim 27 , wherein the antibody remains at least 80% monomeric after incubation for 1 month at room temperature. 
     
     
         38 . The vector of  claim 27 , wherein the antibody is monovalent or multivalent. 
     
     
         39 . The vector of  claim 27 , wherein the antibody is chemically or biologically modified. 
     
     
         40 . The vector of  claim 35 , wherein the antibody is glycosylated, PEGylated or HESylated. 
     
     
         41 . A host cell comprising a heterologous nucleic acid molecule encoding a humanized antibody against IL-1 beta comprising a variable light chain sequence having at least 90% identity to SEQ ID No: 8; and a variable heavy chain sequence having at least 90% identity to SEQ ID No: 12. 
     
     
         42 . The cell of  claim 41 , comprising a variable heavy chain sequence (VH) as set forth in SEQ ID Nos. 9, 10, 11, 12 or 13. 
     
     
         43 . The cell of  claim 41 , comprising a variable light chain (VL) sequence as set forth in SEQ ID Nos. 7, 8 or 24. 
     
     
         44 . The cell of  claim 41 , wherein the antibody has the sequence as set forth in SEQ ID No.: 19. 
     
     
         45 . The cell of  claim 41 , wherein the antibody is an antibody fragment selected from the group consisting of a Fab, a Fab′, a F(ab)′2, a scFv, a Fv fragment, a nanobody, a VHH or a minimal recognition unit. 
     
     
         46 . The cell of  claim 41 , wherein the antibody is a scFv fragment. 
     
     
         47 . A host cell comprising a heterologous nucleic acid molecule encoding a humanized antibody against IL-1 beta comprising variable heavy chain CDR-H1, CDR-H2 and CDR-H3 sequences as set forth in SEQ ID Nos: 4, 21, and 6, respectively, or variants thereof, and wherein
 a) position 24 of the heavy chain is an alanine residue (A);   b) position 25 of the heavy chain is a phenylalanine residue (F);   c) position 44 of the heavy chain is an isoleucine residue (I);   d) position 56 of the heavy chain is a serine residue (S);   e) position 82 of the heavy chain is a lysine residue (K);   f) position 86 of the heavy chain is an arginine residue (R); and/or   g) position 105 of the heavy chain is a phenylalanine residue (F); according to AHo numbering;   and/or comprising variable light chain CDR-L1, CDR-L2 and CDR-L3 sequences as set forth in SEQ ID Nos: 1, 2; and 3, respectively, or variants thereof, and wherein   h) position 1 of the light chain is an aspartic acid residue (D);   i) position 3 of the light chain is a glutamine residue (Q);   j) position 20 of the light chain is a threonine residue (T);   k) position 99 of the light chain is a glutamic acid residue (E);   l) position 105 of the light chain is a phenylalanine residue (F);   m) position 146 of the light chain is a glutamic residue (E);   n) position 147 of the light chain is an isoleucine residue (I);   o) position 148 of the light chain is a lysine residue (K); and/or   p) position 149 of the light chain is an arginine residue (R), according to AHo numbering.   
     
     
         48 . The cell of  claim 47 , wherein the variable heavy chain further comprises at least one of the following residues:
 (i) Serine (S) at heavy chain amino acid position 12 (according to AHo numbering);   (ii) Serine (S) or Threonine (T) at heavy chain amino acid position 103 (according to AHo numbering); and/or   (iii) Serine (S) or Threonine (T) at heavy chain amino acid position 144 (according to AHo numbering).   
     
     
         49 . The cell of  claim 47 , wherein the variable light chain sequence comprises the sequence as set forth in SEQ ID No.: 8 and the variable heavy chain sequence comprises the sequence as set forth in SEQ ID No.: 12. 
     
     
         50 . The cell of  claim 49 , wherein the antibody further comprises a linker sequence set forth in SEQ ID No: 14. 
     
     
         51 . The cell of  claim 47 , wherein the antibody has the sequence as set forth in SEQ ID No.: 19. 
     
     
         52 . The cell of  claim 27 , wherein the antibody is an antibody fragment selected from the group consisting of a Fab, a Fab′, a F(ab)′2, a scFv, a Fv fragment, a nanobody, a VHH or a minimal recognition unit. 
     
     
         53 . The cell of  claim 47 , wherein the antibody is a scFv fragment. 
     
     
         54 . The cell of  claim 47 , wherein the antibody is a full-length immunoglobulin. 
     
     
         55 . The cell of  claim 47 , wherein the antibody has an IC50 with regard to human IL-1 beta of lower than 10 nM wherein the IC50 is determined by inhibiting the IL-1 beta induced release of IL-6 from human fibroblasts. 
     
     
         56 . The cell of  claim 55 , wherein the IC50 with regard to human IL-1 beta is about 5 pM. 
     
     
         57 . The cell of  claim 47 , wherein the antibody remains at least 80% monomeric after incubation for 1 month at room temperature. 
     
     
         58 . The cell of  claim 47 , wherein the antibody is monovalent or multivalent. 
     
     
         59 . The cell of  claim 47 , wherein the antibody is chemically or biologically modified. 
     
     
         60 . The cell of  claim 59 , wherein the antibody is glycosylated, PEGylated or HESylated. 
     
     
         61 . A method of producing an antibody comprising the steps of:
 (i) cultivating the host cell of  claim 41  so that the antibody is expressed;   (ii) recovering the antibody; and   (iii) optionally purifying the antibody.