Anti-transferrin receptor antibodies and methods of use

Abstract

The present invention relates to anti-transferrin receptor antibodies and methods of their use.

1 . An isolated antibody that binds to human transferrin receptor (TfR) and primate TfR, wherein the antibody does not inhibit the binding of transferrin to TfR, wherein the antibody comprises HVR-H1, HVR-H2 and HVR-H3, and HVR-L1, HVR-L2 and HVR-L3, respectively, comprising the amino acid sequences of:
 a) SEQ ID NOs: 53, 54 and 55, and SEQ ID NOs: 50, 51 and 52;   b) SEQ ID NOs: 53, 157 and 55, and SEQ ID NOs: 50, 51 and 52: or   c) SEQ ID NOs: 53, 54 and 55, and SEQ ID NOs: 50, 51 and 155.   
     
     
         2 . An isolated antibody that binds to human TfR and primate TfR, wherein the antibody does not inhibit the binding of transferrin to TfR, and wherein one or more properties of the antibody have been modified to reduce or eliminate the impact of the antibody on reticulocytes and/or reduce the severity or presence of acute clinical symptoms in a subject or mammal treated with the antibody, wherein the antibody comprises HVR-H1, HVR-H2 and HVR-H3, and HVR-L1, HVR-L2 and HVR-L3, respectively, comprising the amino acid sequences of:
 a) SEQ ID NOs: 53, 54 and 55, and SEQ ID NOs: 50, 51 and 52;   b) SEQ ID NOs: 53, 157 and 55, and SEQ ID NOs: 50, 51 and 52: or   c) SEQ ID NOs: 53, 54 and 55, and SEQ ID NOs: 50, 51 and 155.   
     
     
         3 . The antibody of  claim 1 , which is a monoclonal antibody. 
     
     
         4 . The antibody of  claim 1 , which is a human, humanized, or chimeric antibody. 
     
     
         5 . The antibody of  claim 1 , which is an antibody fragment that binds human TfR and primate TfR. 
     
     
         6 .- 10 . (canceled) 
     
     
         11 . The antibody of  claim 1 , comprising (a) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 108 or 154; (b) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 105 or 151 or (c) a VH sequence as in (a) and a VL sequence as in (b). 
     
     
         12 . The antibody of  claim 11 , comprising a VH sequence of
 a. SEQ ID NO: 108;   b. SEQ ID NO: 154.   
     
     
         13 . The antibody of  claim 11 , comprising a VL sequence of
 a. SEQ ID NO:105;   b. SEQ ID NO: 151.   
     
     
         14 . An antibody comprising:
 a. a VH sequence of SEQ ID NO:108 and a VL sequence of SEQ ID NO:105;   b. a VH sequence of SEQ ID NO: 154 and a VL sequence of SEQ ID NO: 105; or   c. a VH sequence of SEQ ID NO: 108 and a VL sequence of SEQ ID NO: 151.   
     
     
         15 .- 20 . (canceled) 
     
     
         21 . The antibody of  claim 2 , wherein the one or more properties are selected from the effector function of the antibody Fc region, the complement activation function of the antibody, the half-life of the antibody and the affinity of the antibody for TfR. 
     
     
         22 . The antibody of  claim 21 , wherein the one or more properties are selected from the effector function of the antibody Fc region and the complement activation function of the antibody, and wherein the effector function or complement activation function has been reduced or eliminated relative to a wild-type antibody of the same isotype. 
     
     
         23 . The antibody of  claim 22 , wherein the effector function is reduced or eliminated by a method selected from reduction of glycosylation of the antibody, modification of the antibody isotype to an isotype that naturally has reduced or eliminated effector function, and modification of the Fc region. 
     
     
         24 . The antibody of  claim 23 , wherein the glycosylation of the antibody is reduced by a method selected from: production of the antibody in an environment that does not permit wild-type glycosylation; removal of carbohydrate groups already present on the antibody; and modification of the antibody such that wild-type glycosylation does not occur. 
     
     
         25 . The antibody of  claim 24 , wherein the antibody is produced in a non-mammalian cell production system, or where the antibody is produced synthetically. 
     
     
         26 . The antibody of  claim 24 , wherein the Fc region of the antibody comprises a mutation at position 297 such that the wild-type asparagine residue at that position is replaced with another amino acid that interferes with glycosylation at that position. 
     
     
         27 . The antibody of  claim 23 , wherein the effector function is reduced or eliminated by at least one modification of the Fc region. 
     
     
         28 . The antibody of  claim 27 , wherein the effector function or complement activation function is reduced or eliminated by deletion of all or a portion of the Fc region, or by engineering the antibody such that it does not include an Fc region or non-Fc region competent for effector function or complement activation function. 
     
     
         29 . The antibody of  claim 27 , wherein the modification is selected from: a point mutation of the Fc region to impair binding to one or more Fc receptors selected from the following positions: 234, 235, 238, 239, 248, 249, 252, 254, 265, 268, 269, 270, 272, 278, 289, 292, 293, 294, 295, 296, 297, 298, 301, 303, 322, 324, 327, 329, 333, 335, 338, 340, 373, 376, 382, 388, 389, 414, 416, 419, 434, 435, 437, 438, and 439; a point mutation of the Fc region to impair binding to C1q selected from the following positions: 270, 322, 329, and 321; eliminating some or all of the Fc region, and a point mutation at position 132 of the CH1 domain. 
     
     
         30 . The antibody of  claim 29 , wherein the modification is at least one point mutation of the Fc region to impair binding to one or more Fc receptors selected from 234, 235, 265, 297 and 329. 
     
     
         31 . The antibody of  claim 30 , wherein the modification is at positions 297 or 265 and 297. 
     
     
         32 . The antibody of  claim 30 , wherein the modification is at positions 234, 235 and 329. 
     
     
         33 . The antibody of  claim 31 , wherein the modification is N297G; D265A and N297A or D265A and N297G. 
     
     
         34 . The antibody of  claim 31 , wherein the modification is L234A, L235A, and P329G. 
     
     
         35 . The antibody of  claim 21 , wherein the one or more properties is the half-life of the antibody. 
     
     
         36 . The antibody of  claim 35  wherein the half-life is increased by a modification in the FcRn binding domain of the antibody at a position selected from: 252, 254, 256, 434 and 436. 
     
     
         37 . The antibody of  claim 36 , wherein the modification is at positions 252, 254 and 256. 
     
     
         38 . The antibody of  claim 36 , wherein the modification is at positions 434 and 436. 
     
     
         39 . The antibody of  claim 37 , wherein the modification is M252Y, S254T and T256E. 
     
     
         40 . The antibody of  claim 38 , wherein the modification is N434A and Y436I. 
     
     
         41 .- 44 . (canceled) 
     
     
         45 . The antibody of  claim 1 , wherein the antibody has a KD or IC50 for TfR of about 1 pM to about 100 μM. 
     
     
         46 . The antibody of  claim 1 , wherein the antibody is coupled to a therapeutic compound. 
     
     
         47 . The antibody of  claim 46 , wherein the antibody is a multispecific antibody and the therapeutic compound optionally forms one portion of the multispecific antibody. 
     
     
         48 . The antibody of  claim 47 , wherein the multispecific antibody comprises a first antigen binding site which binds TfR and a second antigen binding site which binds a brain antigen. 
     
     
         49 . The antibody of  claim 48 , wherein the brain antigen is selected from the group consisting of: beta-secretase 1 (BACE1), Abeta, epidermal growth factor receptor (EGFR), human epidermal growth factor receptor 2 (HER2), tau, apolipoprotein E (ApoE), alpha-synuclein, CD20, huntingtin, prion protein (PrP), leucine rich repeat kinase 2 (LRRK2), parkin, presenilin 1, presenilin 2, gamma secretase, death receptor 6 (DR6), amyloid precursor protein (APP), p75 neurotrophin receptor (p75NTR), and caspase 6. 
     
     
         50 . The antibody of  claim 49 , wherein the multispecific antibody binds both TfR and BACE1. 
     
     
         51 . The antibody of  claim 49 , wherein the multispecific antibody binds both TfR and Abeta. 
     
     
         52 . The antibody of  claim 46 , wherein the therapeutic compound is a neurological disorder drug. 
     
     
         53 . An isolated nucleic acid encoding the antibody of  claim 1 . 
     
     
         54 . A host cell comprising the nucleic acid of  claim 53 . 
     
     
         55 . A method of producing an antibody comprising culturing the host cell of  claim 54  so that the antibody is produced and optionally further comprising recovering the antibody from the host cell. 
     
     
         56 . A pharmaceutical formulation comprising the antibody of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         57 .- 63 . (canceled) 
     
     
         64 . A method of transporting a compound across the BBB in a subject comprising exposing an antibody of  claim 46  to the BBB such that the antibody transports the compound coupled thereto across the BBB. 
     
     
         65 . A method of increasing exposure of the CNS of a subject to a compound, comprising exposing an antibody of  claim 46  to the BBB such that the antibody transports the compound coupled thereto across the BBB. 
     
     
         66 . A method of increasing retention in the CNS of a compound administered to a subject, comprising exposing an antibody of  claim 46  to the BBB such that the retention in the CNS of the compound is increased. 
     
     
         67 . A method of treating a neurological disorder in a mammal comprising treating the mammal with an antibody of  claim 46 . 
     
     
         68 . The method of  claim 67 , wherein the neurological disorder is selected from the group consisting of a neuropathy disorder, a neurodegenerative disease, cancer, an ocular disease disorder, a seizure disorder, a lysosomal storage disease, amyloidosis, a viral or microbial disease, ischemia, a behavioral disorder, and CNS inflammation. 
     
     
         69 . The method of any  claim 64 , wherein the BBB or neurological disorder is in a human subject. 
     
     
         70 . The method of  claim 69 , wherein the dose amount and/or frequency of administration is modulated to reduce the concentration of antibody to which the red blood cells are exposed. 
     
     
         71 . The method of  claim 69 , further comprising the step of monitoring the subject for depletion of red blood cells. 
     
     
         72 . The method of  claim 69 , wherein the antibody coupled to the compound is administered at a therapeutic dose. 
     
     
         73 . The method of  claim 72 , wherein the therapeutic dose is TfR-saturating. 
     
     
         74 . The method of  claim 69 , wherein administration of the antibody is at a dose and/or dose frequency calibrated to minimize acute clinical symptoms of the antibody administration. 
     
     
         75 . The antibody of  claim 14 , wherein the antibody comprises a heavy chain variable region (VH) sequence having the amino acid sequence of SEQ ID NO: 108 and a light chain variable region (VL) sequence having the amino acid sequence of SEQ ID NO: 105. 
     
     
         76 . The antibody of  claim 14 , wherein the antibody comprises a heavy chain variable region (VH) sequence having the amino acid sequence of SEQ ID NO: 154 and a light chain variable region (VL) sequence having the amino acid sequence of SEQ ID NO: 105. 
     
     
         77 . The antibody of  claim 14 , wherein the antibody comprises a heavy chain variable region (VH) sequence having the amino acid sequence of SEQ ID NO: 108 and a light chain variable region (VL) sequence having the amino acid sequence of SEQ ID NO: 151.