BINDING PROTEINS, INCLUDING ANTIBODIES, ANTIBODY DERIVATIVES AND ANTIBODY FRAGMENTS, THAT SPECIFICALLY BIND CD154 AND USES THEREOF
This invention provides binding proteins, including antibodies, antibody derivatives and antibody fragments, that specifically bind a CD154 (CD40L) protein. This invention also provides a chimeric, humanized or fully human antibody, antibody derivative or antibody fragment that specifically binds to an epitope to which a humanized Fab fragment comprising a variable heavy chain sequence according to SEQ ID NO: 1 and comprising a variable light chain sequence according to SEQ ID NO: 2 specifically binds. CD154 binding proteins of this invention may elicit reduced effector function relative to a second anti-CD154 antibody. CD154 binding proteins of this invention are useful in diagnostic and therapeutic methods, such as in the treatment and prevention of diseases including those that involve undesirable immune responses that are mediated by CD154-CD40 interactions.
1 . (canceled)
2 . An anti-CD154 antibody or antigen-binding fragment thereof, comprising:
(a) a V H domain sequence selected from SEQ ID NO: 1, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 11 and an amino acid sequence comprising one or more conservative substitutions that is at least 90% identical to any one of SEQ ID NO: 1, SEQ ID NO: 9, SEQ ID NO: 10 or SEQ ID NO: 11; and a V L domain sequence selected from SEQ ID NO: 2 or SEQ ID NO: 14 and an amino acid sequence comprising one or more conservative substitutions that is at least 90% identical to any one of SEQ ID NO: 2 or SEQ ID NO: 14; or
(b) a V H domain sequence comprising SEQ ID NO: 56 and a V L domain sequence comprising SEQ ID NO: 54;
(c) a V H domain sequence comprising SEQ ID NO: 60 and a V L domain sequence comprising SEQ ID NO: 58;
(d) a V H domain sequence comprising SEQ ID NO: 65 and a V L domain sequence comprising SEQ ID NO: 62;
(e) a V H domain sequence comprising SEQ ID NO: 66 and a V L domain sequence comprising SEQ ID NO: 63;
(f) a V H domain sequence comprising SEQ ID NO: 71 and a V L domain sequence comprising SEQ ID NO: 68;
(g) a V H domain sequence comprising SEQ ID NO: 72 and a V L domain sequence comprising SEQ ID NO: 69.
3 . The anti-CD154 antibody according to claim 2 comprising V H and V L domain sequences selected from:
(a) SEQ ID NO: 1 or an amino acid sequence comprising one or more conservative substitutions that is at least 90% identical to SEQ ID NO: 1 and SEQ ID NO: 2 or an amino acid sequence comprising one or more conservative substitutions that is at least 90% identical to SEQ ID NO: 2, respectively;
(b) SEQ ID NO: 11 or an amino acid sequence comprising one or more conservative substitutions that is at least 90% identical to SEQ ID NO: 11 and SEQ ID NO: 2 or an amino acid sequence comprising one or more conservative substitutions that is at least 90% identical to SEQ ID NO: 2, respectively;
(c) SEQ ID NO: 9 or an amino acid sequence comprising one or more conservative substitutions that is at least 90% identical to SEQ ID NO: 9 and SEQ ID NO: 14 or an amino acid sequence comprising one or more conservative substitutions that is at least 90% identical to SEQ ID NO: 14, respectively;
(d) SEQ ID NO: 10 or an amino acid sequence comprising one or more conservative substitutions that is at least 90% identical to SEQ ID NO: 10 and SEQ ID NO: 14 or an amino acid sequence comprising one or more conservative substitutions that is at least 90% identical to SEQ ID NO: 14, respectively.
4 . The anti-CD154 antibody according to claim 2 , wherein the conservative substitution is not within the CDRs.
5 . The anti-CD154 antibody according to claim 2 , wherein the antibody is selected from a monoclonal antibody, chimeric antibody, primatized antibody and humanized antibody or antigen binding fragments thereof.
6 . The anti-CD154 antibody according to claim 2 , wherein the antibody is selected from a multimeric antibody, heterodimeric antibody, hemidimeric antibody, tetravalent antibody, bispecific antibody and single chain antibody.
7 . The antigen-binding fragment according to claim 2 , wherein the antigen binding fragment is selected from an Fab, F(ab) 2 , Fab′, F(ab′) 2 , F(ab′) 3 , Fd, Fv and a domain antibody.
8 . The anti-CD154 antibody or antigen-binding fragment according to claim 2 , which is modified by a covalent attachment of a functional moiety.
9 . The anti-CD154 antibody or antigen-binding fragment according to claim 8 , wherein the functional moiety is poly(ethyleneglycol) or a derivative thereof.
10 . The anti-CD154 antibody or antigen-binding fragment according to claim 9 , wherein the antigen-binding fragment is a Fab′ wherein a thiol group in a modified hinge region is covalently linked to a maleimide group that is covalently linked to a lysine residue, wherein a methoxypoly(ethyleneglycol) polymer having a molecular weight of approximately 20,000 Da is attached to each of the amine groups of the lysine.
11 . The anti-CD154 antibody according to claim 2 , wherein the antibody comprises an immunoglobulin Fc region selected from IgG1, IgG2, IgG3 and IgG4 Fc regions, or derived from IgG1, IgG2, IgG3 or IgG4 Fc regions.
12 . The anti-CD154 antibody according to claim 2 , wherein the binding protein further comprises a variant Fc region that confers reduced effector function compared to a native or parental Fe region.
13 . The anti-CD154 antibody according to claim 12 , wherein the variant Fe region is a hybrid Fe region comprising sequences from more than one Ig Fc domain type.
14 . The anti-CD154 antibody according to claim 2 , wherein the antibody is not glycosylated.
15 . The anti-CD154 antibody according to claim 12 , wherein the reduced effector function is reduced binding to an Fc receptor (FcR).
16 . The anti-CD154 antibody according to claim 12 , wherein the reduced effector function is reduced binding to a complement protein.
17 . The anti-CD154 antibody according to claim 2 , wherein the antibody or antigen binding fragment is linked to a functional moiety.
18 . An isolated nucleic acid molecule encoding the anti-CD154 antibody of claim 2 .
19 . A vector comprising a nucleic acid molecule of claim 18 .
20 . A host cell comprising a vector of claim 19 .
21 . A method for producing a CD154 binding protein comprising the steps of
(a) culturing a host cell of claim 20 under conditions suitable for expression of the CD154 binding protein by the host cell; and
(b) recovering the CD154 binding protein.
22 . The method according to claim 21 , wherein the host cell is a prokaryotic or a eukaryotic cell.
23 . A composition comprising the anti-CD154 antibody according to claim 2 , or an antigen binding fragment thereof, and a suitable pharmaceutical carrier.
24 . The composition according to claim 23 , further comprising an additional immunosuppressive or immunomodulatory compound or agent.
25 . A method for treating or preventing a human condition, disorder or disease mediated in whole or in part by CD40 signaling, or a symptom of any of the foregoing, the method comprising the step of administering to a subject in need thereof a therapeutically effective amount of the anti-CD154 antibody of claim 2 or an antigen binding fragment thereof.
26 . The method according to claim 25 , wherein the condition, disorder or disease is an inflammatory or autoimmune response or fibrosis.
27 . The method according to claim 26 , wherein the inflammatory or autoimmune response or fibrosis is selected from rheumatoid arthritis, systemic lupus erythematosis, spondyloarthritis, inflammatory bowel disease, Crohn's disease, psoriasis and multiple sclerosis.