ANTI-HtrA1 ANTIBODIES AND METHODS OF USE

Abstract

The invention provides anti-HtrA1 antibodies and methods of using the same.

What is claimed is: 
     
         1 . An isolated antibody that binds to HtrA1 competitively with an antibody comprising a VH sequence of SEQ ID NO:8 and a VL sequence of SEQ ID NO:7. 
     
     
         2 . The antibody of  claim 1 , wherein competitive binding is determined using an ELISA assay. 
     
     
         3 . An isolated antibody that binds to HtrA1, wherein the antibody (i) binds to an epitope that includes N224, K248, or both of HtrA1, and (ii) inhibits HtrA1 with an IC 50  of ≤30 nM. 
     
     
         4 . The antibody of  claim 3 , wherein the antibody further comprises one or more of the following properties: (i) binds to HtrA1 with a ratio of 1 variable domain to one subunit of an HtrA1 trimer, or (ii) does not prevent trimer formation of HtrA1. 
     
     
         5 . The antibody of  claim 3 , wherein the IC 50  is determined using a serine protease assay with a substrate having SEQ ID NO:12. 
     
     
         6 . The antibody of any one of  claims 1 - 5 , wherein the antibody does not cross-react with one or more of HtrA2, HtrA3 and HtrA4. 
     
     
         7 . The antibody of any one of  claims 1 - 6 , wherein the antibody has a dissociation constant of ≤500 nM. 
     
     
         8 . The antibody of  claim 6 , wherein the dissociation constant is determined by BIAcore using a Fab at 25° C. 
     
     
         9 . The antibody of any one of  claims 1 - 8 , which is a monoclonal antibody. 
     
     
         10 . The antibody of any one of  claims 1 - 8 , which is a human, humanized, or chimeric antibody. 
     
     
         11 . The antibody of any one of  claims 1 - 8 , which is an antibody fragment that binds HtrA1. 
     
     
         12 . The antibody of any one of  claims 1 - 11 , wherein the antibody comprises (a) HVR-H3 comprising the amino acid sequence GTFLTX p WGHYFDY, wherein X p  is S or T (SEQ ID NO: 27); (b) HVR-L3 comprising the amino acid sequence QQX g X h X i X j PX k T, wherein X g  is S, V or D; X h  is Y, D or S; X i  is T, S, A, D or N; X j  is T, H, N, S, A, L or R; and X k  is P, T, A or S (SEQ ID NO: 24); and (c) HVR-H2 comprising the amino acid sequence WIDPYGGDTXoYADSVKG, wherein X o  is N or D (SEQ ID NO: 26). 
     
     
         13 . The antibody of any one of  claims 1 - 12 , wherein the antibody comprises (a) HVR-H3 comprising the amino acid sequence of SEQ ID NO:6; (b) HVR-L3 comprising the amino acid sequence of SEQ ID NO:3; and (c) HVR-H2 comprising the amino acid sequence of SEQ ID NO:5. 
     
     
         14 . The antibody of any one of  claims 1 - 12 , wherein the antibody comprises (a) HVR-H3 comprising the amino acid sequence of SEQ ID NO:6; (b) HVR-L3 comprising the amino acid sequence of SEQ ID NO:19; and (c) HVR-H2 comprising the amino acid sequence of SEQ ID NO:5. 
     
     
         15 . The antibody of any one of  claims 1 - 12 , wherein the antibody comprises (a) HVR-H3 comprising the amino acid sequence of SEQ ID NO:6: (b) HVR-L3 comprising the amino acid sequence of SEQ ID NO:22; and (c) HVR-H2 comprising the amino acid sequence of SEQ ID NO:5. 
     
     
         16 . The antibody of any one of  claims 1 - 12 , wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence GFX l IX m X n YYIH, wherein X i  is N, S or T; X m  is S, D, Y or A; and X n  is G or D (SEQ ID NO: 25): (b) HVR-H2 comprising the amino acid sequence WIDPYGGDTX o YADSVKG, wherein X o  is N or D (SEQ ID NO:26); and (c) HVR-H3 comprising the amino acid sequence GTFLTX p WGHYFDY, wherein X p  is S or T (SEQ ID NO: 27). 
     
     
         17 . The antibody of any one of  claims 1 - 12  or  16 , further comprising (a) HVR-L1 comprising the amino acid sequence RASQX a X b X c X d X e X f A, wherein X a  is D, S or V; X b  is V or I; X c  is S, N or G; X d  is T or N; X e  is A or Y; and X f  is V or L (SEQ ID NO: 23); (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (c) HVR-L3 comprising the amino acid sequence QQX g X h X i X j PX k T, wherein X g  is S, V or D; X h  is Y, D or S; X i  is T, S, A, D or N: X j  is T, H, N, S, A, L or R; and X k  is P, T, A or S (SEQ ID NO: 24). 
     
     
         18 . The antibody of any one of  claims 1 - 12 , 16 or 17, wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:4; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:5; and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:6. 
     
     
         19 . The antibody of  claim 18 , further comprising (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:1; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:3. 
     
     
         20 . The antibody of any one of  claims 1 - 12 , 16 or 17, wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:20; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:5; and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:6. 
     
     
         21 . The antibody of  claim 20 , further comprising (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:18; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:19. 
     
     
         22 . The antibody of  claim 20 , further comprising (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:21; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:22. 
     
     
         23 . The antibody of any one of  claims 1 - 12 , 16 or 17, comprising (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:1; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:3. 
     
     
         24 . The antibody of any one of  claims 1 - 12 , 16 or 17, comprising (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:18; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:19. 
     
     
         25 . The antibody of any one of  claims 1 - 2 , 16 or 17, comprising (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:21; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:22. 
     
     
         26 . The antibody of any one of  claims 1 - 25 , further comprising a heavy chain variable domain framework (FR2) sequence of SEQ ID NO:17. 
     
     
         27 . The antibody of any one of  claims 1 - 26 , comprising (a) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:8; (b) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:7; or (c) a VH sequence as in (a) and a VL sequence as in (b). 
     
     
         28 . The antibody of any one of  claims 1 - 27 , comprising (a) a VH sequence comprising SEQ ID NO: 32; (b) a VL sequence comprising SEQ ID NO: 31; or (c) a VH sequence comprising SEQ ID NO: 32 and a VL sequence comprising SEQ ID NO: 31. 
     
     
         29 . The antibody of  claim 27  or  28 , comprising a VH sequence of SEQ ID NO:8. 
     
     
         30 . The antibody of  claim 27  or  28 , comprising a VL sequence of SEQ ID NO:7. 
     
     
         31 . The antibody of  claim 27  or  28 , comprising a VH sequence of SEQ ID NO:30. 
     
     
         32 . The antibody of  claim 27  or  28 , comprising a VL sequence of SEQ ID NO:28. 
     
     
         33 . The antibody of  claim 27  or  28 , comprising a VL sequence of SEQ ID NO:29. 
     
     
         34 . An antibody comprising a VH sequence of SEQ ID NO:8 and a VL sequence of SEQ ID NO:7. 
     
     
         35 . An antibody comprising a VH sequence of SEQ ID NO:30 and a VL sequence of SEQ ID NO:28. 
     
     
         36 . An antibody comprising a VH sequence of SEQ ID NO:30 and a VL sequence of SEQ ID NO:29. 
     
     
         37 . An antibody that binds to HtrA1 comprising (a) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:8; (b) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:7; or (c) a VH sequence as in (a) and a VL sequence as in (b). 
     
     
         38 . An antibody that binds to HtrA1 comprising (a) a VH sequence comprising SEQ ID NO: 32; (b) a VL sequence comprising SEQ ID NO: 31; or (c) a VH sequence comprising SEQ ID NO: 32 and a VL sequence comprising SEQ ID NO: 31. 
     
     
         39 . An antibody comprising (a) HVR-H1 comprising the amino acid sequence GFX l IX m X n YYIH, wherein X l  is N, S or T; X m  is S, D, Y or A; and X n  is G or D (SEQ ID NO: 25); (b) HVR-H2 comprising the amino acid sequence WIDPYGGDTXoYADSVKG, wherein X o  is N or D (SEQ ID NO:26); (c) HVR-H3 comprising the amino acid sequence GTFLTX p WGHYFDY, wherein X p  is S or T (SEQ ID NO: 27); (d) HVR-L1 comprising the amino acid sequence RASQX a X b X c X d X e X f A, wherein X a  is D, S or V; X b  is V or I; X c  is S, N or G; X d  is T or N; X e  is A or Y; and X f  is V or L (SEQ ID NO: 23); (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (f) HVR-L3 comprising the amino acid sequence QQX g X h X i X j PX k T, wherein X g  is S, V or D; X h  is Y, D or S; X i  is T, S, A, D or N; X j  is T, H, N, S, A, L or R; and X k  is P, T, A or S (SEQ ID NO: 24). 
     
     
         40 . The antibody of  claim 39 , wherein the antibody comprises (a) HVR-H1 comprising an amino acid sequence selected from: SEQ ID NO:4, 20, and 47-51; (b) HVR-H2 comprising an amino acid sequence selected from: SEQ ID NO:5 and 52; (c) HVR-H3 comprising an amino acid sequence selected from: SEQ ID NO:6 and 53; (d) HVR-L1 comprising an amino acid sequence selected from: SEQ ID NO:1, 18, 21 and 33; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (f) HVR-L3 comprising an amino acid sequence selected from: SEQ ID NO:3, 19, 22, and 34-46. 
     
     
         41 . The antibody of  claim 40 , wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:4; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:5; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:6; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:1; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:3. 
     
     
         42 . The antibody of  claim 40 , wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:20; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:5; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:6; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:18; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:19. 
     
     
         43 . The antibody of  claim 40 , wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:20; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:5; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:6; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:21; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:22. 
     
     
         44 . The antibody of any one of  claims 1 - 43 , which is a full length IgG1 or IgG4 antibody. 
     
     
         45 . An isolated nucleic acid encoding the antibody of any one of  claims 1 - 44 . 
     
     
         46 . A host cell comprising the nucleic acid of  claim 45 . 
     
     
         47 . A method of producing an antibody comprising culturing the host cell of  claim 46  under conditions suitable for expression of the nucleic acid encoding the anti-HtrA1 antibody. 
     
     
         48 . The method of  claim 47 , further comprising recovering the anti-HtrA1 antibody from the host cell culture. 
     
     
         49 . An immunoconjugate comprising the antibody of any one of  claims 1 - 44  and a cytotoxic agent. 
     
     
         50 . A pharmaceutical formulation comprising the antibody of any one of  claims 1 - 44  and a pharmaceutically acceptable carrier. 
     
     
         51 . The antibody of any one of  claims 1 - 44  for use as a medicament. 
     
     
         52 . The antibody of any one of  claims 1 - 44  for use in treating age-related macular degeneration, geographic atrophy, diabetic retinopathy, retinopathy of prematurity, or polypoidal choroidal vasculopathy. 
     
     
         53 . The antibody of  claim 52 , for use in treating dry age-related macular degeneration. 
     
     
         54 . The antibody of any one of  claims 1 - 44  for use in inhibiting degeneration of retinal or photoreceptor cells. 
     
     
         55 . The antibody of any one of  claims 1 - 44  for use in inhibiting HtrA1 protease activity in an eye. 
     
     
         56 . Use of the antibody of any one of  claims 1 - 44  in the manufacture of a medicament. 
     
     
         57 . The use of  claim 56 , wherein the medicament is for treatment of treating age-related macular degeneration, geographic atrophy, diabetic retinopathy, retinopathy of prematurity, or polypoidal choroidal vasculopathy. 
     
     
         58 . The use of  claim 57 , wherein the age-related macular degeneration is dry age-related macular degeneration. 
     
     
         59 . The use of  claim 56 , wherein the medicament is for inhibiting degeneration of retinal or photoreceptor cells. 
     
     
         60 . The use of  claim 56 , wherein the medicament is for inhibiting HtrA1 protease activity in an eye. 
     
     
         61 . A method of treating an individual having age-related macular degeneration, geographic atrophy, diabetic retinopathy, retinopathy of prematurity, or polypoidal choroidal vasculopathy comprising administering to the individual an effective amount of the antibody of any one of  claims 1 - 44 . 
     
     
         62 . The method of  claim 61 , wherein the age-related macular degeneration is dry age-related macular degeneration. 
     
     
         63 . A method of inhibiting retinal or photoreceptor cell degeneration in an individual comprising administering to the individual an effective amount of the antibody of any one of  claims 1 - 44  to inhibit retinal or photoreceptor cell degeneration. 
     
     
         64 . A method of inhibiting HtrA1 activity in an eye of an individual comprising administering to the individual an effective amount of the antibody of any one of  claims 1 - 44  to inhibit HtrA1 activity in the eye. 
     
     
         65 . The antibody of any one of  claims 1 - 12 , wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 87; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:88; and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 89. 
     
     
         66 . The antibody of any one of  claims 1 - 12  or  65 , further comprising (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 85; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:58; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 86. 
     
     
         67 . An antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 87; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:88; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:89; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:85; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:58; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 86. 
     
     
         68 . The antibody of  claim 67 , wherein the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 25; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:26; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:27; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:23; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:2; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:24. 
     
     
         69 . The antibody of  claim 67  or  68 , wherein the antibody comprises (a) HVR-H1 comprising an amino acid sequence selected from: SEQ ID NO:4, 20, 47-51 and 75-80; (b) HVR-H2 comprising an amino acid sequence selected from: SEQ ID NO:5, 52 and 81-82; (c) HVR-H3 comprising an amino acid sequence selected from: SEQ ID NO:6, 53 and 83-84; (d) HVR-L1 comprising an amino acid sequence selected from: SEQ ID NO:1, 18, 21, 33 and 54-57; (e) HVR-L2 comprising an amino acid sequence selected from: SEQ ID NO:2 and 58; and (f) HVR-L3 comprising an amino acid sequence selected from: SEQ ID NO:3, 19, 22, 34-46 and 59-74. 
     
     
         70 . The antibody of any one of  claims 65 - 69 , which is a full length IgG1 or IgG4 antibody. 
     
     
         71 . An isolated nucleic acid encoding the antibody of any one of  claims 65 - 70 . 
     
     
         72 . A host cell comprising the nucleic acid of  claim 71 . 
     
     
         73 . A method of producing an antibody comprising culturing the host cell of  claim 72  under conditions suitable for expression of the nucleic acid encoding the anti-HtrA1 antibody. 
     
     
         74 . The method of  claim 73 , further comprising recovering the anti-HtrA1 antibody from the host cell culture. 
     
     
         75 . An immunoconjugate comprising the antibody of any one of  claims 65 - 70  and a cytotoxic agent. 
     
     
         76 . A pharmaceutical formulation comprising the antibody of any one of  claims 65 - 70  and a pharmaceutically acceptable carrier. 
     
     
         77 . The antibody of any one of  claims 65 - 70  for use as a medicament. 
     
     
         78 . The antibody of any one of  claims 65 - 70  for use in treating age-related macular degeneration, geographic atrophy, diabetic retinopathy, retinopathy of prematurity, or polypoidal choroidal vasculopathy. 
     
     
         79 . The antibody of  claim 78 , for use in treating dry age-related macular degeneration. 
     
     
         80 . The antibody of any one of  claims 65 - 70  for use in inhibiting degeneration of retinal or photoreceptor cells. 
     
     
         81 . The antibody of any one of  claims 65 - 70  for use in inhibiting HtrA1 protease activity in an eye. 
     
     
         82 . Use of the antibody of any one of  claims 65 - 70  in the manufacture of a medicament. 
     
     
         83 . The use of  claim 82 , wherein the medicament is for treatment of treating age-related macular degeneration, geographic atrophy, diabetic retinopathy, retinopathy of prematurity, or polypoidal choroidal vasculopathy. 
     
     
         84 . The use of  claim 83 , wherein the age-related macular degeneration is dry age-related macular degeneration. 
     
     
         85 . The use of  claim 82 , wherein the medicament is for inhibiting degeneration of retinal or photoreceptor cells. 
     
     
         86 . The use of  claim 82 , wherein the medicament is for inhibiting HtrA1 protease activity in an eye. 
     
     
         87 . A method of treating an individual having age-related macular degeneration, geographic atrophy, diabetic retinopathy, retinopathy of prematurity, or polypoidal choroidal vasculopathy comprising administering to the individual an effective amount of the antibody of any one of  claims 65 - 70 . 
     
     
         88 . The method of  claim 87 , wherein the age-related macular degeneration is dry age-related macular degeneration. 
     
     
         89 . A method of inhibiting retinal or photoreceptor cell degeneration in an individual comprising administering to the individual an effective amount of the antibody of any one of  claims 65 - 70  to inhibit retinal or photoreceptor cell degeneration. 
     
     
         90 . A method of inhibiting HtrA1 activity in an eye of an individual comprising administering to the individual an effective amount of the antibody of any one of  claims 65 - 70  to inhibit HtrA1 activity in the eye.