Antibodies to cd40

Abstract

The present invention relates to antibodies and antigen-binding portions thereof that specifically bind to CD40, preferably human CD40, and that function as CD40 agonists. The invention also relates to human anti-CD40 antibodies and antigen-binding portions thereof. The invention also relates to antibodies that are chimeric, bispecific, derivatized, single chain antibodies or portions of fusion proteins. The invention also relates to isolated heavy and light chain immunoglobulins derived from human anti-CD40 antibodies and nucleic acid molecules encoding such immunoglobulins. The present invention also relates to methods of making human anti-CD40 antibodies, compositions comprising these antibodies and methods of using the antibodies and compositions for diagnosis and treatment. The invention also provides gene therapy methods using nucleic acid molecules encoding the heavy and/or light immunoglobulin molecules that comprise the human anti-CD40 antibodies. The invention also relates to transgenic animals comprising nucleic acid molecules of the present invention.

What is claimed is: 
     
         1 . A monoclonal antibody or an antigen-binding portion thereof that specifically binds to and activates human CD40, wherein said antibody or antigen-binding portion comprises:
 a) a heavy chain comprising amino acid sequences of a heavy chain CDR1, a heavy chain CDR2 and a heavy chain CDR3 from a heavy chain variable region;
 wherein said amino acid sequences of the heavy chain CDR1 and the heavy chain CDR2 are independently selected from a CDR1 and a CDR2 of a heavy chain variable region, respectively, wherein the sequence of said heavy chain variable region comprises no more than 18 amino acid changes from the amino acid sequence encoded by a germline V H  3-30+, 4-59, 1-02, 4.35 or 3-30.3 gene; 
 wherein said amino acid sequence of the heavy chain CDR3 is from a CDR3 of a heavy chain variable region, wherein said heavy chain variable region is selected from the group consisting of 
   i) a heavy chain variable region of an antibody selected from the group consisting of 3.1.1, 3.1.1H-A78T, 3.1.1H-A78T-V88A-V97A, 7.1.2, 10.8.3, 15.1.1, 21.4.1, 21.2.1, 22.1.1, 22.1.1H-C109A, 23.5.1, 23.25.1, 23.28.1, 23.28.1H-D16E, 23.29.1 and 24.2.1;   ii) a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 2, 10, 18, 26, 34, 42, 50, 58, 66, 74 and 82, or a heavy chain variable region comprising an amino acid sequence from the heavy chain amino acid sequence selected from the group consisting of SEQ ID NOS: 90, 92, 96 and 98, or said amino acid sequence lacking a signal sequence; and   iii) a heavy chain variable region encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NOS: 1, 9, 17, 25, 33, 41, 49, 57, 65, 73 and 81, or a heavy chain variable region comprising an amino acid sequence encoded by the heavy chain nucleic acid sequence selected from the group consisting of 89, 91, 95 and 97, or said encoded heavy chain variable region lacking a signal sequence;   wherein the amino acid sequence of said heavy chain CDR3 may have up to two conservative amino acid substitutes and/or two non-conservative amino acid insertions, deletions or substitutions therefrom; or   b) a light chain comprising amino acid sequences of a light chain CDR1, a light chain CDR2 and a light chain CDR3 from a light chain variable region,
 wherein said amino acid sequences of the light chain CDR1 and the light chain CDR3 are independently selected from CDR1 and a CDR3 of a light chain variable region, respectively, wherein the light chain variable region comprises no more than ten amino acid changes from the amino acid sequence encoded by a germline Vκ A3/A19, L5 or A27 gene; and 
 wherein said amino acid sequence of the light chain CDR2 is from a light chain variable region, wherein said light chain variable region is selected from the group consisting of 
   i) a light chain variable region of an antibody selected from the group consisting of 3.1.1, 3.1.1L-L4M-L83V, 7.1.2, 10.8.3, 15.1.1, 21.4.1, 21.2.1, 22.1.1, 23.5.1, 23.25.1, 23.28.1, 23.28.1L-C92A, 23.29.1 and 24.2.1;   ii) a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 4, 12, 20, 28, 36, 44, 52, 60, 68, 76 and 84, or a light chain variable region comprising an amino acid sequence from the light chain amino acid sequence selected from the group consisting of SEQ ID NOS: 94 and 100, or said amino acid sequence lacking a signal sequence; and   iii) a light chain variable region encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NOS: 3, 11, 19, 27, 35, 43, 51, 59, 67, 75 and 83, or a light chain variable region comprising an amino acid sequence encoded by the light chain nucleic sequence selected from the group consisting of SEQ ID NOS: 93 and 99, or said encoded light chain variable region lacking a signal sequence;   wherein the amino acid sequence of said light chain CDR2 may have up to two conservative amino acid substitutes and/or two non-conservative amino acid insertions, deletions or substitutions therefrom.   
     
     
         2 . The antibody or antigen-binding portion thereof according to  claim 1 , wherein the antibody or portion thereof comprises a heavy chain and a light chain, and wherein the amino acid sequences of the heavy chain variable region of said heavy chain and the light chain variable region of said light chain are respectively selected from the group consisting of:
 a) the amino acid sequence selected from the group consisting of SEQ ID NOS: 2, 90 and 92, and the amino acid sequence selected from the group consisting of SEQ ID NOS: 4 and 94;   b) the amino acid sequence of SEQ ID NO: 10 and the amino acid sequence of SEQ ID NO: 12;   c) the amino acid sequence of SEQ ID NO: 18 and the amino acid sequence of SEQ ID NO: 20;   d) the amino acid sequence of SEQ ID NO: 26 and the amino acid sequence of SEQ ID NO: 28;   e) the amino acid sequence of SEQ ID NO: 34 and the amino acid sequence of SEQ ID NO: 36;   f) the amino acid sequence of SEQ ID NO: 42 and the amino acid sequence of SEQ ID NO: 44;   g) the amino acid sequence selected from the group consisting of SEQ ID NOS: 50 and 96 and the amino acid sequence of SEQ ID NO: 52;   h) the amino acid sequence of SEQ ID NO: 58 and the amino acid sequence of SEQ ID NO: 60;   i) the amino acid sequence selected from the group consisting of SEQ ID NOS: 66 and 98, and the amino acid sequence selected from the group consisting of SEQ ID NOS: 68 and 100; and   j) the amino acid sequence of SEQ ID NO: 74 and the amino acid sequence of SEQ ID NO: 78;   wherein said amino acid sequences optionally lack a signal sequence and wherein the amino acid sequences of the heavy and light chains may each have up to six conservative amino acid substitutions and/or four non-conservative amino acid insertions, deletions or substitutions therefrom.   
     
     
         3 . The antibody or antigen-binding portion according to  claim 1 , wherein the amino acid sequences of the heavy chain and the light chain are respectively selected from the group consisting of:
 a) the amino acid sequence of SEQ ID NO: 6 and the amino acid sequence of SEQ ID NO: 8;   b) the amino acid sequence of SEQ ID NO: 14 and the amino acid sequence of SEQ ID NO: 16;   c) the amino acid sequence of SEQ ID NO: 22 and the amino acid sequence of SEQ ID NO: 24;   d) the amino acid sequence of SEQ ID NO: 30 and the amino acid sequence of SEQ ID NO: 32; and   e) the amino acid sequence of SEQ ID NO: 46 and the amino acid sequence of SEQ ID NO: 48.   
     
     
         4 . The antibody or antigen-binding portion according to  claim 1 , wherein the antibody or portion has at least one property selected from the group consisting of:
 a) does not bind to mouse, rat, dog and/or rabbit B cells;   b) binds to human, cynomolgus and/or rhesus B cells;   c) has a selectivity for CD40 that is at least 100 times greater than its selectivity for receptor activator of nuclear factor-kappa B (RANK), 4-1BB (CD137), tumor necrosis factor receptor-1 (TNFR-1) and tumor necrosis factor receptor-2 (TNFR-2);   d) binds to CD40 with a K D  of 4×10 −10  M or less;   e) has an off rate for CD40 of K off  of 2×10 −4  or smaller;   f) inhibits tumor growth in vivo in the presence of human T cells and/or human dendritic cells;   g) inhibits the growth of CD40-positive tumors in the absence of human immune cells;   h) increases expression of ICAM, MHC-II, B7-2, CD71, CD23 and/or CD71 on the surface of human B-cells;   i) increases secretion of IL-12p40, IL-12p70 and/or IL-8 by human dendritic cells;   j) increases expression of ICAM, MHC-II, B7-2 and/or CD83 on the surface of human dendritic cells;   k) increases expression of interferon-gamma by human T cells during allogenic stimulation;   l) binds human CD40 in the presence of human CD40L; and   m) binds to an epitope of human CD40 contained in domain 1, domain 2, or domain 3 of the extracellular domain of CD40.   
     
     
         5 . A monoclonal antibody or an antigen-binding portion thereof that specifically binds to and activates human CD40, wherein the antibody or portion has at least one property selected from the group consisting of:
 a) cross-competes for binding to CD40 with an antibody selected from the group consisting of 3.1.1, 3.1.1H-A78T, 3.1.1H-A78T-V88A-V97A, 3.1.1L-L4M-L83V, 7.1.2, 10.8.3, 15.1.1, 21.4.1, 21.2.1, 22.1.1, 22.1.1H-C109A, 23.5.1, 23.25.1, 23.28.1, 23.28.1L-C92A, 23.28.1H-D16E, 23.28.1L-C92A, 23.29.1, 23.29.1L-R174K and 24.2.1;   b) binds to the same epitope of CD40 as an antibody selected from the group consisting of 3.1.1, 3.1.1H-A78T, 3.1.1H-A78T-V88A-V97A, 3.1.1L-L4M-L83V, 7.1.2, 10.8.3, 15.1.1, 21.4.1, 21.2.1, 22.1.1, 22.1.1H-C109A, 23.5.1, 23.25.1, 23.28.1, 23.28.1L-C92A, 23.28.1H-D16E, 23.28.1L-C92A, 23.29.1, 23.29.1L-R174K and 24.2.1;   c) binds to CD40 with substantially the same K D  as an antibody selected from the group consisting of 3.1.1, 3.1.1H-A78T, 3.1.1H-A78T-V88A-V97A, 3.1.1L-L4M-L83V, 7.1.2, 10.8.3, 15.1.1, 21.4.1, 21.2.1, 22.1.1, 22.1.1H-C109A, 23.5.1, 23.25.1, 23.28.1, 23.28.1L-C92A, 23.28.1H-D16E, 23.28.1L-C92A, 23.29.1, 23.29.1L-R174K and 24.2.1; and   d) binds to CD40 with substantially the same off rate as an antibody selected from the group consisting of 3.1.1, 3.1.1H-A78T, 3.1.1H-A78T-V88A-V97A, 3.1.1L-L4M-L83V, 7.1.2, 10.8.3, 15.1.1, 21.4.1, 21.2.1, 22.1.1, 22.1.1H-C109A, 23.5.1, 23.25.1, 23.28.1, 23.28.1L-C92A, 23.28.1H-D16E, 23.28.1L-C92A, 23.29.1, 23.29.1L-R174K and 24.2.1.   
     
     
         6 . The antibody or antigen-binding portion thereof according to  claim 1  that is
 a) an immunoglobulin G (IgG), an IgM, an IgE, an IgA or an IgD molecule, or is derived therefrom; or 
 b) an Fab fragment, an F(ab′) 2  fragment, an F v  fragment, a single chain antibody, a humanized antibody, a chimeric antibody or a bispecific antibody. 
 
     
     
         7 . A pharmaceutical composition comprising the antibody or antigen-binding portion according to  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         8 . A method of treating a patient in need thereof with an anti-CD40 antibody or an antigen-binding portion thereof, comprising the step of administering to the patient an effective amount of the antibody or portion according to  claim 1 . 
     
     
         9 . A method of treating cancer in a patient in need thereof with an anti-CD40 antibody or an antigen-binding portion thereof, comprising the step of administering to the patient an effective amount of the antibody or portion according to  claim 1 . 
     
     
         10 . The method according to  claim 9 , further comprising the step of administering to the patient radiotherapy, chemotherapy, photodynamic therapy, or surgery. 
     
     
         11 . The method according to  claim 9 , further comprising the step of administering to the patient an anti-cancer agent. 
     
     
         12 . A method of enhancing an immune response in a patient in need thereof with an anti-CD40 antibody or an antigen-binding portion thereof, comprising the step of administering to the patient an effective amount of the antibody or portion according to  claim 1 . 
     
     
         13 . An isolated cell line that produces the antibody or antigen-binding portion according to  claim 1 . 
     
     
         14 . An isolated nucleic acid molecule that comprises a nucleic acid sequence that encodes the heavy chain or an antigen-binding portion thereof or the light chain or an antigen-binding portion thereof of the antibody according to  claim 1 . 
     
     
         15 . A vector comprising the nucleic acid molecule according to  claim 14 , wherein the vector optionally comprises an expression control sequence operably linked to the nucleic acid molecule. 
     
     
         16 . A host cell comprising the nucleic acid molecule according to  claim 14 . 
     
     
         17 . A method of making an anti-CD40 antibody or an antigen-binding portion thereof, comprising culturing the host cell according to  claim 16  under suitable conditions and recovering said antibody or antigen-binding portion. 
     
     
         18 . A non-human transgenic animal or transgenic plant comprising the nucleic acid according to  claim 14 , wherein the non-human transgenic animal or transgenic plant expresses said nucleic acid. 
     
     
         19 . A method of making an antibody or an antigen-binding portion thereof that specifically binds to human CD40, comprising the step of isolating the antibody or antigen-binding portion from the non-human transgenic animal or transgenic plant according to  claim 18 . 
     
     
         20 . A method of treating a subject in need thereof with an antibody or an antigen-binding portion thereof that specifically binds to and activates human CD40, comprising the steps of:
 (a) administering an effective amount of an isolated nucleic acid molecule encoding the heavy chain or an antigen-binding portion thereof, an isolated nucleic acid molecule encoding the light chain or an antigen-binding portion thereof, or both the nucleic acid molecules encoding the light chain and the heavy chain or antigen-binding portions thereof; and   (b) expressing the nucleic acid molecule(s).