US2008003218A1PendingUtilityA1

Anti-IgE antibodies

59
Assignee: GENENTECH INCPriority: Jul 2, 1997Filed: Apr 11, 2006Published: Jan 3, 2008
Est. expiryJul 2, 2017(expired)· nominal 20-yr term from priority
Inventors:Henry B. LowmanLeonard G. PrestaPaula M. JardieuJohn Lowe
A61P 37/00C07K 2317/56C07K 2317/565C07K 16/4291C07K 2317/76A61K 39/39566A61P 11/06C07K 2317/55
59
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Claims

Abstract

The present invention relates to a method for adjusting the affinity of a polypeptide to a target molecule by a combination of steps, including: (1) the identification of aspartyl residues which are prone to isomerization; (2) the substitution of alternative residues and screening the resulting mutants for affinity against the target molecule. In a preferred embodiment, the method of subtituting residues is affinity maturation with phage display (AMPD). In a further preferred embodiment the polypeptide is an antibody and the target molecule is an antigen. In a further preferred embodiment, the antibody is anti-IgE and the target molecule is IgE. In another embodiment, the invention relates to an anti-IgE antibody having improved affinity to IgE.

Claims

exact text as granted — not AI-modified
1 - 65 . (canceled)  
     
     
         66 . An antibody molecule comprising an e26 sequence selected from the group consisting of: Fab fragment (SEQ ID NOs: 19-20), sFv fragment (SEQ ID NO: 22) and F(ab′) 2  (SEQ ID NOs: 24-25).  
     
     
         67 . An antibody molecule having the sequence “e26” of  FIG. 12  (SEQ ID NOs:15-16).  
     
     
         68 . An antibody molecule comprising an e27 sequence selected from the group consisting of: Fab fragment (SEQ ID NOs: 19 and 21), sFv fragment (SEQ ID NO: 23) and F(ab′) 2  (SEQ ID NOs: 24 and 26).  
     
     
         69 . An antibody molecule having the sequence “e27” of  FIG. 12  (SEQ ID NO:17-18).  
     
     
         70 . A composition comprising pharmaceutically-acceptable excipient(s) in admixture with an e26 antibody molecule having a sequence selected from the group consisting of: Fab (SEQ ID NOs: 19-20); sFv (SEQ ID NO: 22) and F(ab′) 2  (SEQ ID NOs: 24-25).  
     
     
         71 . A composition comprising pharmaceutically-acceptable excipient(s) in admixture with an antibody having the sequence “e26” of  FIG. 12  (SEQ ID NOs: 15-16).  
     
     
         72 . A composition comprising pharmaceutically-acceptable excipient(s) in admixture with an e27 antibody molecule having a sequence selected from the group consisting of: Fab (SEQ ID NOs: 19 and 20); sFv (SEQ ID NO: 23) and F(ab′) 2  (SEQ ID NOs: 24 and 26).  
     
     
         73 . A composition comprising pharmaceutically-acceptable excipient(s) in admixture with an antibody having the sequence “e27” of  FIG. 12  (SEQ ID NOs: 17-18).  
     
     
         74 . An improved anti-IgE antibody which does not exhibit aspartic acid isomerization produced by: 
 (a) identifying aspartyl residues prone to isomerization in an unimproved anti-IgE antibody or antibody fragment;    (b) substituting alternative residues in lieu of the aspartyl residues to create candidate molecules and screening the resulting candidate molecules for affinity against IgE; and    (c) selecting those candidates which display affinity equal to or greater than the unimproved anti-IgE antibody.    
     
     
         75 . The improved antibody of  claim 74  wherein the unimproved anti-IgE antibody is E-25.  
     
     
         76 . The improved antibody of  claim 75  wherein the method of substituting residues is affinity maturation with phage display (AMPD).  
     
     
         77 . A method of inhibiting the IgE mediated release of histamine from mast cells or basophils in a mammal comprising the administration of a therapeutically effective amount of an E26 antibody fragment comprising a sequence selected from the group consisting of: F(ab) (SEQ ID NO: 19-20); sFv (SEQ ID NO: 22) and F(ab′) 2  (SEQ ID NO: 24-25).  
     
     
         78 . The method of  claim 77  wherein the administered E26 antibody fragment comprises an F(ab) fragment (SEQ ID NOs: 19 and 20).  
     
     
         79 . The method of  claim 77  wherein the administered E26 antibody fragment comprises an sFv fragment (SEQ ID NO:22).  
     
     
         80 . The method of  claim 77  wherein the administered E26 antibody fragment comprises an F(ab′) 2  fragment (SEQ ID NOs: 24 and 25).  
     
     
         81 . A method of inhibiting the IgE mediated release of histamine from mast cells or basophils in a mammal comprising the administration of a therapeutically effective amount of an E27 antibody comprising the variable light and heavy chain sequences (SEQ ID NOs: 17-18).  
     
     
         82 . A method of treating a disorder mediated by IgE comprising the administration to a mammal in need thereof a therapeutically effective amount of an E26 antibody sequence fragment selected from the group consisting of: F(ab)(SEQ ID NOs: 19-20); sFv (SEQ ID NO: 22) and F(ab′) 2  (SEQ ID NOs: 24-25).  
     
     
         83 . The method of  claim 82  wherein the administered E26 antibody fragment comprises an F(ab) fragment (SEQ ID NOs: 19 and 20).  
     
     
         84 . The method of  claim 82  wherein the administered E26 antibody fragment comprises an sFv fragment (SEQ ID NO:22).  
     
     
         85 . The method of  claim 82  wherein the administered E26 antibody fragment comprises an F(ab′) 2  fragment (SEQ ID NOs: 24 and 25).  
     
     
         86 . A method of treating a disorder mediated by IgE comprising the administration to a mammal in need thereof a therapeutically effective amount of E27 antibody molecule comprising the variable light and heavy chain sequences (SEQ ID NOs: 17 and 18).  
     
     
         87 . A method of inhibiting the IgE mediated release of histamine from mast cells or basophils in a mammal comprising the administration of a therapeutically effective amount of an E27 antibody fragment comprising a sequence selected from the group consisting of: an F(ab) fragment (SEQ ID NOs: 19 and 21); an sFv fragment (SEQ ID NO: 23); and an F(ab′)2 fragment (SEQ ID NOs: 24 and 26).  
     
     
         88 . The method of  claim 87  wherein the E27 antibody fragment comprises an F(ab) fragment (SEQ ID NOs: 19 and 21).  
     
     
         89 . The method of  claim 87  wherein the E27 antibody fragment comprises an sFv fragment (SEQ ID NO: 23).  
     
     
         90 . The method of  claim 87  wherein the E27 antibody fragment comprises an F(ab′) 2  fragment (SEQ ID NOs: 24 and 26).  
     
     
         91 . A method of treating a disorder mediated by IgE comprising the administration to a mammal in need thereof a therapeutically effective amount of an E27 antibody fragment comprising a sequence selected from the group consisting of: an F(ab) fragment (SEQ ID NOs: 19 and 21); an sFv fragment (SEQ ID NO: 23); and an F(ab′) 2  fragment (SEQ ID NOs: 24 and 26).  
     
     
         92 . The method of  claim 91  wherein the administered E27 antibody fragment comprises an F(ab) fragment (SEQ ID NOs: 19 and 21).  
     
     
         93 . The method of  claim 91  wherein the administered E27 antibody fragment comprises an sFv fragment (SEQ ID NO: 23).  
     
     
         94 . The method of  claim 91  wherein the administered E27 antibody fragment comprises an F(ab′) 2  fragment (SEQ ID NOs: 24 and 26).  
     
     
         95 . A method of inhibiting the IgE mediated release of histamine from mast cells or basophils in a mammal comprising the administration of a therapeutically effective amount of an improved anti-IgE antibody or antibody fragment, wherein said anti-IgE antibody or antibody fragment is produced by: (a) identifying aspartyl residues prone to isomerization in an unimproved anti-IgE antibody or antibody fragment; (b) substituting alternative residues in lieu of the aspartyl residues to create candidate molecules for affinity against IgE; and (c) selecting those candidates which display affinity equal to or greater than the unimproved anti-IgE antibody or antibody fragment.  
     
     
         96 . A humanized antibody comprising the variable light chain and heavy chain domain of the antibody “E26” of  FIG. 12  (SEQ ID NOS:15 and 16).  
     
     
         97 . The antibody of  claim 96  further comprising an IgG1 constant region.  
     
     
         98 . A composition comprising the antibody of  claim 97  and a pharmaceutically-acceptable carrier.  
     
     
         99 . The antibody of  claim 96  further comprising an IgG2 constant region.  
     
     
         100 . A composition comprising the antibody of  claim 99  and a pharmaceutically-acceptable carrier.  
     
     
         101 . The antibody of  claim 96  further comprising an IgG3 constant region.  
     
     
         102 . A composition comprising the antibody of  claim 101  and a pharmaceutically-acceptable carrier.  
     
     
         103 . The antibody of  claim 96  further comprising an IgG4 constant region.  
     
     
         104 . A composition comprising the antibody of  claim 103  and a pharmaceutically-acceptable carrier.  
     
     
         105 . A humanized antibody comprising the variable light chain and heavy chain domain of the antibody “E27” of  FIG. 12  (SEQ ID NOS:17 and 18).  
     
     
         106 . The humanized antibody of  claim 105  further comprising an IgG1 constant region.  
     
     
         107 . A composition comprising the antibody of  claim 106  and a pharmaceutically-acceptable carrier.  
     
     
         108 . The humanized antibody of  claim 105  further comprising an IgG2 constant region.  
     
     
         109 . A composition comprising the antibody of  claim 108  and a pharmaceutically-acceptable carrier.  
     
     
         110 . The humanized antibody of  claim 105  further comprising an IgG3 constant region.  
     
     
         111 . A composition comprising the antibody of  claim 110  and a pharmaceutically-acceptable carrier.  
     
     
         112 . The humanized antibody of  claim 105  comprising an IgG4 constant region.  
     
     
         113 . A composition comprising the antibody of  claim 112  and a pharmaceutically-acceptable carrier.  
     
     
         114 . An antibody comprising the heavy and light chain sequences of “E26” in  FIG. 12  (SEQ ID NOs:15-16).  
     
     
         115 . An article of manufacture, comprising the antibody of  claim 114  in a container in combination with a label, wherein said label indicates that said antibody is useful for treating an IgE-mediated disorder.  
     
     
         116 . An antibody comprising the heavy and light chain sequences of “E27” in  FIG. 12  (SEQ ID NOs:17-18).  
     
     
         117 . An article of manufacture, comprising the antibody of  claim 116  in a container in combination with a label, wherein said label indicates that said antibody is useful for treating an IgE-mediated disorder.  
     
     
         118 . A composition comprising pharmaceutically-acceptable excipient(s) in admixture with an E26 antibody molecule comprising the heavy and light chain sequences of  FIG. 12  (SEQ ID NOs:15-16).  
     
     
         119 . An article of manufacture, comprising the composition of  claim 118  in a container in combination with a label, wherein said label indicates that said composition is useful for treating an IgE-mediated disorder.  
     
     
         120 . A composition comprising pharmaceutically-acceptable excipient(s) in admixture with an E27 antibody molecule comprising the heavy and light chain sequences of  FIG. 12  (SEQ ID NOs:17-18).  
     
     
         121 . An article of manufacture, comprising the composition of  claim 120  in a container in combination with a label, wherein said label indicates that said composition is useful for treating an IgE-mediated disorder.  
     
     
         122 . The improved anti-IgE antibody or IgE binding fragment thereof comprising a sequence selected from the group consisting of: (a) the heavy and light variable domain sequences of E26 (SEQ ID NOs:15-16); (b) the heavy and light variable domain sequences of E27 (SEQ ID NOs:17-18); and (c) IgE binding fragments of (a) or (b).  
     
     
         123 . An article of manufacture, comprising the antibody of  claim 122  in a container in combination with a label, wherein said label indicates that said antibody is useful for treating an IgE-mediated disorder.  
     
     
         124 . The improved anti-IgE antibody of  claim 122 , comprising the heavy and light variable domain sequences of E26 (SEQ ID NOs:15-16).  
     
     
         125 . An article of manufacture, comprising the antibody of  claim 124  in a container in combination with a label, wherein said label indicates that said antibody is useful for treating an IgE-mediated disorder.  
     
     
         126 . The improved anti-IgE antibody of  claim 124 , comprising the heavy and light variable domain sequences of E27 (SEQ D NOs:17-18).  
     
     
         127 . An article of manufacture, comprising the antibody of  claim 126  in a container in combination with a label, wherein said label indicates that said antibody is useful for treating an IgE-mediated disorder.  
     
     
         128 . The improved IgE binding antibody fragment of  claim 122 , comprising an IgE binding fragment of the heavy and light variable domain sequences of E26 (SEQ ID NOs:15-16).  
     
     
         129 . The improved IgE binding antibody fragment of  claim 122 , comprising an IgE binding fragment of the heavy and light variable domain sequences of E27 (SEQ ID NOs:17-18).  
     
     
         130 . A composition of an improved anti-IgE antibody or IgE binding fragment thereof in combination with an adjunct immunosuppressive agent, wherein the improved anti-IgE antibody or IgE binding fragment thereof comprises: (a) the heavy and light chains of E26 (SEQ ID NOs:15-16); (b) the heavy and light chains of E27 (SEQ ID NOs:17-18); or (c) antigen binding fragments of (a) or (b).  
     
     
         131 . The composition of  claim 130 , wherein the immunosuppressive agent is selected from the group consisting of azathioprine, cyclophosphamide, bromocryptine and glutaraldehyde.  
     
     
         132 . The composition of  claim 130 , wherein the immunosuppressive agent is a glucocorticosteroid.  
     
     
         133 . The composition of  claim 132 , wherein the immunosuppressive agent is selected from the group consisting of: prednisone, methylprednisone and dexamethasone.  
     
     
         134 . The composition of  claim 130 , wherein the immunosuppressive agent is selected from the group consisting of: cyclosporin A; an anti-CD3 antibody; an anti-CD4 antibody and an anti-CD4a antibody.  
     
     
         135 . The composition of  claim 130 , wherein the immunosuppressive agent is selected from the group consisting of: a soluble peptide containing an LFA-3 binding domain; streptokinase; TGF-β; streptodomase; deoxyspergualin, rapamycin and a T-cell receptor.  
     
     
         136 . The composition of  claim 130 , wherein the improved anti-IgE antibody comprises the heavy and light chain sequences of E26 (SEQ ID NOs:15-16).  
     
     
         137 . The composition of  claim 130 , wherein the improved IgE binding fragment comprises an E26 sequence selected from the group consisting of: an Fab fragment (SEQ ID NOs:19-20; an sFv fragment (SEQ ID NO:22) and an F(ab′) 2  fragment (SEQ ID NOs: 24-25).  
     
     
         138 . The composition of  claim 130 , wherein the improved IgE binding fragment comprises an E27 sequence selected from the group consisting of: an Fab fragment (SEQ ID NOs: 19 and 21); an sFv fragment (SEQ ID NO:23) and an F(ab′) 2  fragment (SEQ ID NOs: 24 and 26).  
     
     
         139 . The composition of  claim 130 , wherein the improved anti-IgE antibody comprises the heavy and light chains of E27 (SEQ ID NOs: 17-18).  
     
     
         140 . The composition of  claim 130 , wherein the immunosuppressive agent is a 2-amino-5-aryl-substituted pyrimidine.  
     
     
         141 . The composition of  claim 130 , wherein the immunosuppressive agent is an anti-idiotypic antibody which binds MHC antigens or fragments.  
     
     
         142 . The composition of  claim 130 , wherein the immunosuppressive agent is a T-cell receptor antibody.  
     
     
         143 . The composition of  claim 130 , wherein the immunosuppressive agent is a cytokine antagonist or a cytokine receptor antagonist.  
     
     
         144 . The composition of  claim 143 , wherein the immunosuppressive agent is an antibody selected from the group consisting of: an anti-interferon-γ antibody, an anti-interferon-β; antibody, an anti-interferon-α antibody; an anti-tumor necrosis factor-α antibody, an anti-tumor necrosis factor-β antibody, an anti-interleukin-2 antibody, anti-IL-2 receptor antibody and an anti-L3T4 antibody.  
     
     
         145 . An anti-IgE antibody or IgE binding fragment comprising a variable heavy chain and a variable light chain, wherein the variable light chain further comprises a CDR-L1 of the sequence RASKPVDGEGDSYLNWY (SEQ ID NO:45).  
     
     
         146 . The antibody or IgE binding fragment of  claim 145  further comprises a CDR-H2 of the sequence: SIKYSGETKYNPSVKG (SEQ ID NO:50).  
     
     
         147 . An anti-IgE antibody or IgE binding antibody fragment comprising a variable heavy chain and variable light chain, wherein: 
 (a) the variable light chain further comprises a CDR-L1, a CDR-L2 and a CDR-L3; and    (b) the variable heavy chain further comprises a CDR-H1, a CDR-H2 and a CDR-H3; 
 wherein further:  
 (i) CDR-L1 consists of the sequence: RASKPVDGEGDSYLNWY (SEQ ID NO:45);  
 (ii) CDR-L2 consists of the sequence: AASYLES (SEQ ID NO:46);  
 (iii) CDR-L3 consists of the sequence: QQSHEDPY (SEQ ID NO:47);  
 (iv) CDR-H1 consists of the sequence: GYSITSGYSWN (SEQ ID NO:48);  
 (v) CDR-H2 consists of the sequence: SITYDGSTNYNPSVKG (SEQ ID NO:49) or SIKYSGETKYNPSVKG (SEQ ID NO:50);  
 (vi) CDR-H3 consists of the sequence: GSHYFGHWHFAV (SEQ ID NO:51).  
   
     
     
         148 . The antibody of  claim 147  wherein CDR-H2 is SITYDGSTNYNPSVKG or SIKYSGETKYNPSVKG.  
     
     
         149 . The antibody of  claim 148  wherein CDR-H2 is SIKYSGETKYNPSVKG.  
     
     
         150 . The antibody of  claim 148  further comprising a constant region selected from the group consisting of: IgG1, IgG2, IgG3 and IgG4.  
     
     
         151 . The antibody of  claim 147  wherein the constant region is IgG1.  
     
     
         152 . The antibody of  claim 147  which is multispecific.  
     
     
         153 . The antibody of  claim 147  which is bispecific.  
     
     
         154 . The antibody or antibody fragment of  claim 147  which is an immunoconjugate.  
     
     
         155 . A composition comprising the antibody of  claim 151  in combination with at least one pharmaceutically-acceptable carrier, excipient or stabilizer.  
     
     
         156 . An article of manufacture comprising the composition of  claim 155  in a container in combination with a label, wherein the label indicates that the composition is useful for treating an IgE-mediated disorder.  
     
     
         157 . The article of  claim 156 , wherein the container is a vial.  
     
     
         158 . The article of  claim 156 , wherein the container is a syringe.  
     
     
         159 . The article of  claim 156 , wherein the IgE mediated disorder is selected from the group consisting of: hypersensitivity, asthma, rhinitis, conjunctivitis, eczema, urticaria and food allergy.  
     
     
         160 . A method of treating an IgE-mediated disorder comprising administering to a mammal in need thereof at a therapeutically effective amount of the antibody of  claim 151 .  
     
     
         161 . The method of  claim 160 , wherein the IgE-mediated disorder is selected from the group consisting of: hypersensitivity, asthma, rhinitis, conjunctivitis, eczema, urticaria and food allergy.  
     
     
         162 . An isolated nucleic acid molecule encoding the anti-IgE antibody or IgE binding fragment of  claim 147 .  
     
     
         163 . A vector comprising the nucleic acid of  claim 162 .  
     
     
         164 . The vector of  claim 163  operably linked to control sequences recognized by a host cell transformed with the vector.  
     
     
         165 . A host cell comprising the vector of  claim 164 .  
     
     
         166 . The host cell of  claim 165  which is prokaryotic cell.  
     
     
         167 . The host cell of  claim 165  which is a yeast cell.  
     
     
         168 . The host cell of  claim 165  which is eukaryotic cell.  
     
     
         169 . The host cell of  claim 168  which is a Chinese hamster ovary (CHO) cell.  
     
     
         170 . A process for producing an anti-IgE antibody, IgE-binding fragment or polypeptide comprising culturing the host cell of  claim 165  under conditions suitable for expression of nucleic acid encoding the anti-IgE antibody or IgE-binding fragment, and recovering the anti-IgE antibody or IgE-binding fragment from the cell culture.

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