Antibody polypeptides that antagonize cd40l

Abstract

Antibody polypeptides that specifically bind human CD40L are provided. The antibody polypeptides do not activate platelets. The antibody polypeptides are useful in the treatment of diseases involving CD40L activation, such as graft-related diseases and autoimmune diseases. The antibody polypeptides may be domain antibodies (dAbs) comprising a single VH or VK domain. The half-life of the antibody polypeptides may be increased by modifying the antibody polypeptides to be dual specific reagents that can also bind human serum albumin (HSA) or another antigen.

1 : An isolated antibody polypeptide comprising a first variable domain that specifically binds human CD40L, wherein CD40L comprises the amino acid sequence of SEQ ID NO: 1, wherein the amino acid sequence of the first variable domain comprises
 (a) a CDR1 region which differs from the CDR1 region of BMS2h-572-633 by up to three amino acids,   (b) a CDR2 region which differs from the CDR2 region of BMS2h-572-633 by up to three amino acids, and   (c) a CDR3 region which differs from the CDR3 region of BMS2h-572-633 by up to three amino acids.   
     
     
         2 . (canceled) 
     
     
         3 : The antibody polypeptide of  claim 1 , wherein the amino acid sequence of the first variable domain differs from the amino acid sequence of BMS2h-572-633 by up to 10 amino acids. 
     
     
         4 - 6 . (canceled) 
     
     
         7 : The antibody polypeptide of  claim 1 , wherein the antibody polypeptide is selected from the lineage group of BMS2h-572, and wherein the amino acid sequence of the first variable domain further comprises:
 (a) a CDR1 region having a sequence Trp-X1-Leu-Met-Gly (SEQ ID NO: 2), wherein X1 is Glu or Gln;   (b) a CDR2 region having a sequence Gly-Ile-Glu-Gly-Pro-Gly-Asp-Val-Thr-Tyr-Tyr-Ala-Asp-Ser-Val-Lys-Gly (SEQ ID NO: 3); and   (c) a CDR3 region having a sequence Lys-X2-Y2-Z2-Ser-Asp-Tyr (SEQ ID NO: 4), wherein X2 is Asp or Glu, Y2 is Ala or Ser, and Z2 is Lys, Asn, or Arg.   
     
     
         8 . (canceled) 
     
     
         9 : The antibody polypeptide of  claim 1 , wherein the first variable domain comprises the amino acid sequence of BMS2h-572-633. 
     
     
         10 : The antibody polypeptide of  claim 1 , wherein the antibody polypeptide is a domain antibody (dAb). 
     
     
         11 : The antibody polypeptide of  claim 10 , wherein the antibody polypeptide is a fusion polypeptide comprising the first variable domain and an Fc domain. 
     
     
         12 : The fusion polypeptide of  claim 11 , wherein the fusion polypeptide comprises an IgG4 Fc domain. 
     
     
         13 : The fusion polypeptide of  claim 11 , wherein the fusion polypeptide comprises an IgG1 Fc domain. 
     
     
         14 - 20 . (canceled) 
     
     
         21 : A nucleic acid encoding the antibody polypeptide of  claim 11 . 
     
     
         22 : A vector comprising the nucleic acid of  claim 21 . 
     
     
         23 : An isolated host cell comprising the vector of  claim 22 . 
     
     
         24 : A pharmaceutical composition comprising a therapeutically-effective amount of the antibody polypeptide of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         25 . (canceled) 
     
     
         26 : A method of treating an immune disease in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of the pharmaceutical composition of  claim 24 . 
     
     
         27 - 37 . (canceled) 
     
     
         38 : An isolated antibody polypeptide comprising a first variable domain, wherein said antibody polypeptide specifically binds human CD40L, wherein CD40L comprises the amino acid sequence of SEQ ID NO: 1, wherein the antibody polypeptide competes with the binding of BMS2h-572-633, and wherein the antibody polypeptide inhibits binding of CD40L to CD40 with an EC50 of 100 pM to 100 nM. 
     
     
         39 : The isolated antibody polypeptide of  claim 38 , wherein the first variable domain comprises the amino acid sequence of one of the antibody polypeptides selected from the lineage group consisting of BMS2h-572, BMS2h-719, BMS2h-503, and BMS2h-116. 
     
     
         40 : The isolated antibody polypeptide of  claim 39 , wherein the first variable domain comprises an amino acid sequence at least 95% identical to BMS2h-572-6, BMS2h-572-608, BMS2h-572-614, BMS2h-572-619, BMS2h-572-633, BMS2h-572-634, BMS2h-572-635, BMS2h-719-2, BMS2h-719-202, BMS2h-719-203, BMS2h-719-213, BMS2h-719-214, BMS2h-719-215, BMS2h-719-218, BMS2h-719-225, BMS2h-503-1, BMS2h-503-2, BMS2h-116-1312, BMS2h-116-1313, or BMS2h-116-1320. 
     
     
         41 . (canceled) 
     
     
         42 : The nucleic acid of  claim 21 , wherein the antibody polypeptide comprises a fusion of
 (i) a single variable domain comprising three complementarity determining regions (CDR) and that specifically binds human CD40L, and   (ii) an Fc domain,   
       wherein
 a) CDR1 of the single variable domain has the same amino acid sequence as amino acids 31 to 35 of BMS2h-572-633 (SEQ ID NO: 274); 
 b) CDR2 of the single variable domain has the same amino acid sequence as amino acids 50 to 66 of BMS2h-572-633 (SEQ ID NO: 274); and 
 c) CDR3 of the single variable domain has the same amino acid sequence as amino acids 101 to 107 of BMS2h-572-633 (SEQ ID NO: 274).