Humanized Anti-OX40 Antibodies and Uses Thereof

Abstract

The disclosure provides humanized anti-OX40 antibodies. Also provided are methods of making such antibodies, and methods of use, e.g., treatment of cancer.

1 - 70 . (cancelled) 
     
     
         71 . An antibody or antigen-binding fragment thereof, comprising:
 a humanized heavy chain variable region (VH) and a humanized light chain variable region (VL);   wherein the VH comprises an amino acid sequence with the formula:
   HFW1-HCDR1-HFW2-HCDR2-HFW3-HCDR3-HFW4, 
   wherein HFW 1 is SEQ ID NO: 6 or SEQ ID NO: 7, HCDR1 is SEQ ID NO: 8, HFW2 is SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, or SEQ ID NO: 13, HCDR2 is SEQ ID NO: 14, SEQ ID NO: 15 or SEQ ID NO: 16, HFW3 is SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, or SEQ ID NO: 24, HCDR3 is SEQ ID NO: 25, SEQ ID NO: 26, or SEQ ID NO: 27, and HFW4 is SEQ ID NO: 28;   wherein the VL comprises the amino acid sequence SEQ ID NO: 29 or SEQ ID NO: 32; and   wherein the antibody or antigen-binding fragment thereof can specifically bind to human OX40.   
     
     
         72 . The antibody or antigen-binding fragment thereof of  claim 71 , wherein the VH comprises the amino acid sequence of SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, or SEQ ID NO: 67. 
     
     
         73 . The antibody or antigen-binding fragment thereof of  claim 71  further comprising a light chain constant region or fragment thereof fused to the C-terminus of the VL, wherein the light chain constant region is a human kappa constant region. 
     
     
         74 . The antibody or antigen-binding fragment thereof of  claim 71 , further comprising a heavy chain constant region or fragment thereof fused to the C-terminus of the VH, wherein the heavy chain constant region is a human IgG1 constant region, a human IgG4P constant region, a human IgG1TM constant region, or a murine IgG1 constant region. 
     
     
         75 . The antibody or antigen-binding fragment thereof of  claim 71  further comprising the heavy chain amino acid sequence SEQ ID NO: 71 and the light chain amino acid sequence SEQ ID NO: 30. 
     
     
         76 . The antibody or antigen-binding fragment of  claim 71 , wherein the antigen-binding fragment is an Fv fragment, an Fab fragment, an F(ab′)2 fragment, an Fab′ fragment, a dsFv fragment, an scFv fragment, or an sc(Fv)2 fragment, or any combination thereof. 
     
     
         77 . The antibody or antigen-binding fragment thereof of  claim 71 , which can specifically bind to OX40 as expressed on Jurkat cells, primary activated CD4+ or CDS+ T cells from human, cynomolgus monkey, rhesus monkey, or any combination thereof and which does not bind to murine or rat OX40. 
     
     
         78 . The antibody or antigen-binding fragment thereof of  claim 71 , which does not cross react with related Tumor Necrosis Factor Receptor Superfamily (TNFRSF) proteins. 
     
     
         79 . The antibody or antigen-binding fragment thereof of  claim 71 , which can induce dose-dependent proliferation of activated CD4+ T cells and dose-dependent cytokine release in primary activated human CD4+ T cells in a plate-based assay. 
     
     
         80 . The antibody or antigen-binding fragment thereof of  claim 79 , wherein the cytokine is IFNγ, TNFα, IL-5, IL-10, IL-2, IL-4, IL-13, IL-8, IL-12 p70, IL-1, or any combination thereof. 
     
     
         81 . The antibody or antigen-binding fragment thereof of  claim 71 , which can activate the NFKB pathway in OX40 expressing T cells in the presence of FcγR-expressing cells. 
     
     
         82 . The antibody or antigen-binding fragment thereof of  claim 71 , which can trigger complement-dependent or antibody-dependent cellular cytotoxicity against OX40-expressing cells. 
     
     
         83 . The antibody or antigen-binding fragment thereof of  claim 71 , wherein administration of an effective dose to a subject in need of cancer treatment can inhibit tumor growth in the subject. 
     
     
         84 . The antibody or antigen-binding fragment thereof of  claim 83 , wherein the tumor growth inhibition is achieved in the presence of T cells. 
     
     
         85 . The antibody or antigen-binding fragment thereof of  claim 84 , wherein tumor growth is inhibited by at least 10%, at least 20%, at least 30%, at least 40%, and least 50%, at least 60%, or at least 70% compared to administration of an isotype-matched control antibody or antigen-binding fragment thereof. 
     
     
         86 . A composition comprising the antibody or antigen-binding fragment thereof of  claim 71  and a carrier. 
     
     
         87 . A polynucleotide comprising a nucleic acid that encodes the antibody or antigen-binding fragment thereof of  claim 71 . 
     
     
         88 . The polynucleotide of  claim 87 , wherein the polynucleotide comprises the nucleic acid of SEQ ID NO: 60, the nucleic acid of SEQ ID NO: 31, the nucleic acid of SEQ ID NO: 72, or any combination thereof. 
     
     
         89 . A vector comprising the polynucleotide of  claim 87 . 
     
     
         90 . A host cell comprising the polynucleotide of  claim 87 . 
     
     
         91 . A method of producing an antibody or antigen-binding fragment thereof, comprising culturing the host cell of  claim 90  under conditions in which the antibody or antigen-binding fragment thereof encoded by the polynucleotide is expressed; and recovering the antibody or antigen-binding fragment thereof. 
     
     
         92 . A method to promote survival or proliferation of activated T cells, comprising contacting activated T cells with the antibody or antigen-binding fragment thereof of  claim 71 , wherein the antibody or antigen-binding fragment thereof can specifically bind to OX40 on the surface of the T cells, wherein the activated T cells are activated CD4+ T cells, activated CDS+T cells, or a combination thereof. 
     
     
         93 . A method of inducing cytokine release from activated T cells, comprising contacting activated T cells with the antibody or antigen-binding fragment thereof of  claim 71 , wherein the antibody or antigen-binding fragment thereof can specifically bind to OX40 on the surface of the T cells; and releasing a cytokine from the activated T cells. 
     
     
         94 . The method of  claim 93 , wherein the cytokine is IFNγ, TNFα, IL-5, IL-10, IL-2, IL-4, IL-13, IL-8, IL-12 p70, IL-1, or any combination thereof. 
     
     
         95 . A method of promoting T cell activation, comprising contacting T cells with the antibody or antigen-binding fragment thereof of  claim 71 , wherein the antibody or antigen-binding fragment thereof can specifically bind to OX40 on the surface of the T cells. 
     
     
         96 . The method of  claim 95 , wherein T cell activation can be measured through stimulation of the NFKB signal transduction pathway. 
     
     
         97 . The method of  claim 95 , wherein the T cells are activated CD4+ T cells, activated CDS+ T cells, or a combination thereof. 
     
     
         98 . The method of  claim 97 , wherein the activated CD4+ T cells are human CD4+ T cells, cynomolgus monkey CD4+ T cells, rhesus monkey CD4+ T cells, or a combination thereof. 
     
     
         99 . The method of  claim 95 , wherein the contacting comprises administering an effective amount of the antibody or antigen-binding fragment thereof to a subject. 
     
     
         100 . A method of treating cancer in a subject, comprising administering to a subject in need of treatment an effective amount of the composition of  claim 86 . 
     
     
         101 . The method of  claim 100 , wherein the cancer is a solid tumor. 
     
     
         102 . The method of  claim 101 , wherein administration of the composition can inhibit tumor growth, can promote tumor reduction, or both. 
     
     
         103 . The method of  claim 102 , wherein tumor growth inhibition is achieved in the presence of T cells. 
     
     
         104 . A method of enhancing an immune response in a subject comprising administering to a subject in need thereof a therapeutically effective amount of the antibody or antigen-binding fragment thereof of  claim 71 . 
     
     
         105 . The antibody or antigen-binding fragment thereof of  claim 71 , which binds to an epitope of human OX40 that falls within amino acids 108 to 146 of SEQ ID NO: 91. 
     
     
         106 . The antibody or antigen-binding fragment thereof of  claim 105 , wherein the epitope comprises at least amino acids leucine 116 and alanine 126 of SEQ ID NO: 91. 
     
     
         107 . The antibody or antigen-binding fragment thereof of  claim 71 , which binds to a mouse OX40 variant comprising SEQ ID NO: 92, except for a Q113L mutation and a V124A mutation. 
     
     
         108 . An isolated peptide consisting of 100 or fewer amino acids and comprising amino acids 108 to 146 of SEQ ID NO: 91 except for one, two, three, four, five, or six single amino acid substitutions, deletions, or insertions at any position except L116 and A126, wherein the peptide can be specifically bound by the antibody or antigen-binding fragment thereof of  claim 71 .