US2014154255A1PendingUtilityA1
Anti-vegf antibodies and their uses
Est. expiryNov 30, 2032(~6.4 yrs left)· nominal 20-yr term from priority
Inventors:Yoshiko Akamatsu
A61P 37/02A61P 35/00A61P 35/04A61P 9/10A61P 3/10A61P 27/02C07K 16/22Y02P20/582C07K 2317/565C07K 2317/92C07K 2317/24A61P 19/02C07K 2317/41C07K 2317/72C07K 2317/31A61K 47/6845C07K 16/28C07K 2317/52C07K 2317/526C07K 2317/524C07K 2317/732
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Claims
Abstract
The present disclosure relates to antibodies directed to vascular endothelial growth factor (“VEGF”) and uses of such antibodies, for example to treat diseases associated with the activity and/or overproduction of VEGF.
Claims
exact text as granted — not AI-modifiedWhat is claimed is:
1 . An anti-VEGF antibody or an anti-VEGF binding fragment of an antibody which comprises a VEGF heavy chain variable region comprising CDRs having amino acid sequences corresponding to SEQ ID NO:3 (CDR-H1), SEQ ID NO:4 (CDR-H2), and SEQ ID NO:5 (CDR-H3), and a VEGF light chain variable region comprising CDRs having amino acid sequences corresponding SEQ ID NO:6 (CDR-L1), SEQ ID NO:7 (CDR-L2) and SEQ ID NO:8, wherein CDR-H1 and/or CDR-H3 comprises one or more of the substitutions in Table 21, and optionally one or more mutations or combinations of mutations selected from one or more of Tables 6, 7, 8, 9, 19, 12-1 to 12-9, 13-16, 21, or 22, wherein the six CDRs altogether have up to 17 amino acid substitutions as compared to CDR sequences of the antibody bevacizumab or the antibody ranibizumab.
2 . The anti-VEGF antibody or anti VEGF binding fragment of claim 1 which comprises one or more substitutions selected from T30K; T30N, N31H; N31L; N31W; N31Y; H 97 F; H97Y; S100aQ; and S100aT.
3 . The anti-VEGF antibody or anti VEGF binding fragment of claim 1 which comprises one or more substitutions selected from T28G; T28R; T28Y; T30R; and S100aA.
4 . The anti-VEGF antibody or anti VEGF binding fragment of claim 1 which comprises one or more substitutions selected from N31F; N31H; and N31Y.
5 . The anti-VEGF antibody or anti VEGF binding fragment of claim 1 which comprises one or more substitutions selected T30N; N31F; N31H, N31W and N31Y; and S100aT.
6 . The anti-VEGF antibody or an anti-VEGF binding fragment of claim 1 which comprises the substitution T30K.
7 . The anti-VEGF antibody or an anti-VEGF binding fragment of claim 1 which comprises the substitution T30N.
8 . The anti-VEGF antibody or an anti-VEGF binding fragment of claim 1 which comprises the substitution N31H.
9 . The anti-VEGF antibody or an anti-VEGF binding fragment of claim 1 which comprises the substitution N31L.
10 . The anti-VEGF antibody or an anti-VEGF binding fragment of claim 1 which comprises the substitution N31W.
11 . The anti-VEGF antibody or an anti-VEGF binding fragment of claim 1 which comprises the substitution N31Y.
12 . The anti-VEGF antibody or an anti-VEGF binding fragment of claim 1 which comprises the substitution H97F.
13 . The anti-VEGF antibody or an anti-VEGF binding fragment of claim 1 which comprises the substitution H97Y.
14 . The anti-VEGF antibody or an anti-VEGF binding fragment of claim 1 which comprises the substitution S100aQ.
15 . The anti-VEGF antibody or an anti-VEGF binding fragment of claim 1 which comprises the substitution S100aT.
16 . The antibody of any of claims 1 to 15 , which is a DVD-Ig in which said VEGF heavy and light chain variable regions together form a VEGF binding portion and which further comprises a second binding portion, said second binding portion comprising second heavy chain and light chain variable regions which together bind to a second target.
17 . A DVD-Ig comprising:
(a) a VEGF binding portion comprising a VEGF heavy chain variable region comprising CDRs having amino acid sequences corresponding to SEQ ID NO:3 (CDR-H1), SEQ ID NO:4 (CDR-H2), and SEQ ID NO:5 (CDR-H3) and a VEGF light chain variable region comprising CDRs having amino acid sequences corresponding SEQ ID NO:6 (CDR-L1), SEQ ID NO:7 (CDR-L2) and SEQ ID NO:8, wherein CDRs of the VEGF binding portion include one or more substitutions as compared to bevacizumab or ranibizumab selected from N31F in CDR-H1; K64S in CDR-H2; K64Q in CDR-H2; Y53F in CDR-H2; H97E in CDR-H3; H97D in CDR-H3; H97P in CDR-H3; Y98F in CDR-H3; Y99E in CDR-H3; Y99D in CDR-H3; S100aG in CDR-H3; and T51A in CDR-L2, wherein the anti-VEGF binding portion optionally has one or more additional mutations or combinations of mutations selected from one or more of Tables 6, 7, 8, 9, 10, 11, 12-1 to 12-9, 13-16, or 21-22, wherein the six CDRs of the VEGF binding portion altogether have up to 17 amino acid substitutions as compared to CDR sequences of the antibody bevacizumab or the antibody ranibizumab; and (b) a second binding portion comprising second heavy and light chain variable regions which together bind to a second target.
18 . The antibody of claim 16 or claim 17 , wherein the second binding portion is on the outside of the VEGF binding portion.
19 . The antibody of claim 16 or claim 17 , wherein the VEGF binding portion is on the outside of the second binding portion.
20 . The antibody of any one of claims 16 to 19 , wherein the VEGF and second heavy chain variable regions are connected via a short linker, optionally ASTKGP (SEQ ID NO:427) or GGGGSG (SEQ ID NO:430).
21 . The antibody of any one of claims 16 to 19 , wherein the VEGF and second heavy chain variable regions are connected via a medium linker, optionally GGGGSGGGGS (SEQ ID NO:431).
22 . The antibody of any one of claims 16 to 19 , wherein the VEGF and second heavy chain variable regions are connected via a long linker, optionally ASTKGPSVFPLAP (SEQ ID NO:429) or GGGGSGGGGSGGGG (SEQ ID NO:432).
23 . The antibody of any one of claims 16 to 22 , wherein the VEGF and second light chain variable regions are connected via a short linker, optionally TVAAP (SEQ ID NO: 418), QPKAAP (SEQ ID NO: 421), or GGSGG (SEQ ID NO: 423).
24 . The antibody of any one of claims 16 to 22 , wherein the VEGF and second light chain variable regions are connected via a medium linker, optionally GGSGGGGSG (SEQ ID NO:424).
25 . The antibody of any one of claims 16 to 22 , wherein the VEGF and second light chain variable regions are connected via a long linker, optionally TVAAPSVFIFPP (SEQ ID NO:420), QPKAAPSVTLFPP (SEQ ID NO:422), or GGSGGGGSGGGGS (SEQ ID NO:425).
26 . The antibody of any one of claims 16 to 25 , wherein the second target is DLL4.
27 . The antibody of claim 26 , wherein the CDRs second heavy and light chain variable regions have the CDR sequences of the CDRs of DLL4 binding portions SEQ ID NO:413 and SEQ ID NO:414.
28 . The antibody of claim 27 , wherein the second heavy and light chain variable regions have the sequences of the DLL4 binding portions of SEQ ID NO:413 and SEQ ID NO:414.
29 . The antibody or binding fragment of any one of claims 1 - 28 , wherein CDR-H2 includes the substitution K64S.
30 . The antibody or binding fragment of any one of claims 1 - 28 , wherein CDR-H2 includes the substitution K64Q.
31 . The antibody or binding fragment of any one of claims 1 - 28 , wherein CDR-H2 includes the substitutions Y53F and K64Q.
32 . The antibody or binding fragment of any one of claims 1 - 12 and 14 - 31 , wherein CDR-H3 includes the substitutions H97E and Y98F.
33 . The antibody or binding fragment of any one of claims 1 - 32 , wherein CDR-L2 includes the substitution T51A, and wherein CDR-L2 in said VEGF antibody does not consist of a CDR-L2 sequence set forth in Table 13.
34 . The antibody or binding fragment of any one of claims 1 - 7 and 12 - 33 , wherein said CDR-H1 includes the substitution N31F, and wherein CDR-H1 in said VEGF antibody does not consist of a CDR-H1 sequence set forth in Tables 12-1 to 12-9.
35 . The antibody or binding fragment of any one of claims 1 - 11 and 14 - 34 , wherein said CDR-H3 includes the substitution H97E, and wherein CDR-H3 in said VEGF antibody does not consist of a CDR-H3 sequence set forth in Tables 12-1 to 12-9.
36 . The antibody or binding fragment of any one of claims 1 - 11 , 14 - 31 , and 33 - 34 , wherein said CDR-H3 includes the substitution H97D, and wherein CDR-H3 in said VEGF antibody does not consist of a CDR-H3 sequence set forth in Tables 12-1 to 12-9.
37 . The antibody or binding fragment of any one of claims 1 - 11 , 14 - 31 , and 33 - 34 , wherein said CDR-H3 includes the substitution H97P, and wherein CDR-H3 in said VEGF antibody does not consist of a CDR-H3 sequence set forth in Tables 12-1 to 12-9.
38 . The antibody or binding fragment of any one of claims 1 - 37 , wherein said CDR-H3 includes the substitution Y99E, and wherein CDR-H3 in said VEGF antibody does not consist of a CDR-H3 sequence set forth in Tables 12-1 to 12-9.
39 . The antibody or binding fragment of any one of claims 1 - 37 , wherein said CDR-H3 includes the substitution Y99D, and wherein CDR-H3 in said VEGF antibody does not consist of a CDR-H3 sequence set forth in Tables 12-1 to 12-9.
40 . The antibody or binding fragment of any one of claims 1 - 13 and 18 - 39 , wherein said CDR-H3 includes the substitution S100aG, and wherein CDR-H3 in said VEGF antibody does not consist of a CDR-H3 sequence set forth in Tables 12-1 to 12-9.
41 . The antibody or binding fragment of any one of claims 1 - 13 , and 18 - 40 , wherein said CDR-H3 includes the substitution S100aG, and wherein position 3 in CDR-H3 of said VEGF antibody is not tyrosine.
42 . The antibody or binding fragment of any one of claims 1 - 41 , wherein CDR-H1 includes at least one substitution selected from T28P, N31F, N31G and N31M, and wherein CDR-H1 in said VEGF antibody does not consist of a CDR-H1 sequence set forth in Tables 12-1 to 12-9.
43 . The antibody or binding fragment of any one of claims 1 - 42 , wherein CDR-H3 includes at least one substitution selected from H97A, H97Q, H97S, H97T, S100aD, S100aE, and S100aV, and wherein CDR-H3 in said VEGF antibody does not consist of a CDR-H3 sequence set forth in Tables 12-1 to 12-9.
44 . The antibody or binding fragment of any one of claims 8 - 43 , wherein CDR-H1 includes the substitution T30W, T30R or T30Q, and wherein CDR-H1 in said VEGF antibody does not consist of a CDR-H1 sequence set forth in Tables 12-1 to 12-9.
45 . The antibody or binding fragment of any one of claims 1 - 44 , wherein CDR-H2 includes at least one substitution selected from Y53F, T58F, A61G, A61K, A61R, A61H, A61Y, K64G, K64E, R65L, R65T, R65A, R65E, and R65D, and wherein CDR-H2 in said VEGF antibody does not consist of a CDR-H2 sequence set forth in Tables 12-1 to 12-9.
46 . The antibody or binding fragment of any one of claims 1 - 45 , wherein CDR-H3 includes at least one substitution selected from Y98F and Y100eF, and wherein CDR-H3 in said VEGF antibody does not consist of a CDR-H3 sequence set forth in Tables 12-1 to 12-9.
47 . The antibody or binding fragment of any one of claims 1 - 18 , wherein the heavy chain CDRs include a combination of substitutions selected from:
(a) N31F in CDR-H1, H97D in CDR-H3, Y99D in CDR-H3, and S100aG in CDR-H3; (b) N31F in CDR-H1, H97P in CDR-H3, Y99D in CDR-H3, and S100aG in CDR-H3; (c) N31F in CDR-H1, H97P in CDR-H3, and Y99E in CDR-H3; (d) N31F in CDR-H1, H97E in CDR-H3, and Y99E in CDR-H3; (e) N31F in CDR-H1, H97D in CDR-H3, and Y99E in CDR-H3; (f) N31F in CDR-H1, H97E in CDR-H3, Y99D in CDR-H3, and S100aG in CDR-H3; (g) N31F in CDR-H1, Y99D in CDR-H3, and S100aG in CDR-H3; (h) N31F in CDR-H1, H97P in CDR-H3, and Y99D in CDR-H3; (i) N31F in CDR-H1, H97D in CDR-H3, and S100aG in CDR-H3; (j) N31F in CDR-H1 and S100aG in CDR-H3; (k) N31F in CDR-H1, H97P in CDR-H3, and S100aG in CDR-H3.
48 . The anti-VEGF antibody or anti-VEGF binding fragment of any one of claims 1 to 47 , wherein the six CDRs altogether have up to 16, up to 15, up to 14, up to 13, up to 12, up to 11, up to 10, up to 9, up to 8, up to 7, up to 6, up to 5, or up to 4 amino acid substitutions as compared to CDR sequences of the antibody bevacizumab or the antibody ranibizumab.
49 . The anti-VEGF antibody or anti-VEGF binding fragment of any one of claims 1 to 46 , wherein any individual CDR has no more than three amino acid substitutions as compared to the corresponding CDR sequence of the antibody bevacizumab or the antibody ranibizumab.
50 . The anti-VEGF antibody or anti-VEGF binding fragment of any one of claims 1 to 46 , wherein any individual CDR has no more than two amino acid substitutions as compared to the corresponding CDR sequence of the antibody bevacizumab or the antibody ranibizumab.
51 . The anti-VEGF antibody or anti-VEGF binding fragment of any one of claims 1 to 50 which is a monoclonal antibody or anti-VEGF binding fragment of a monoclonal antibody, respectively.
52 . The antibody or binding fragment of any one of claims 1 to 51 which is a human or humanized antibody, or anti-VEGF binding fragment of a humanized or humanized antibody, respectively.
53 . The antibody or binding fragment of any one of claims 1 to 52 which is an IgG.
54 . The antibody or binding fragment of claim 53 which is an IgG 1 .
55 . The antibody or binding fragment of claim 53 which is an IgG 2 .
56 . The antibody or binding fragment of any one of claims 1 to 55 which is a dual-variable domain (DVD) immunoglobulin.
57 . The antibody or binding fragment of claim 56 which has an anti-DLL4 variable binding domain.
58 . The antibody or binding fragment of any one of claims 1 to 57 which includes one or more mutations in the Fc region that increase ADCC activity.
59 . The antibody or binding fragment of any one of claims 1 to 58 which is non-fucosylated.
60 . The antibody or binding fragment of any one of claims 1 to 59 which includes one or more mutations in the Fc region that increase binding to FcγR.
61 . The antibody or binding fragment of any one of claims 1 to 59 which includes one or more mutations in the Fc region that increase binding to FcRn.
62 . The antibody or binding fragment of any one of claims 1 to 54 which includes one or more mutations in the Fc region that decrease ADCC activity.
63 . The antibody or binding fragment of any one of claims 1 to 62 which has, other than said one or more mutations, a V H sequence corresponding to SEQ ID NO:1 and a V L sequence corresponding to SEQ ID NO:2.
64 . The antibody or binding fragment of any one of claims 1 - 15 , 29 - 55 , and 58 - 63 which has, other than said one or more mutations, a heavy chain sequence corresponding to SEQ ID NO:9 and a light chain sequence corresponding to SEQ ID NO:10.
65 . The antibody or binding fragment of any one of claims 1 to 63 which has an affinity that is 1.5- to 50-fold greater than the affinity of an antibody having a V H sequence corresponding to SEQ ID NO:1 and a V L sequence corresponding to SEQ ID NO:2.
66 . The antibody or binding fragment of any one of claims 1 to 63 which has an affinity that is 1.5- to 50-fold greater than the affinity of an antibody having a heavy chain sequence corresponding to SEQ ID NO:9 and a light chain sequence corresponding to SEQ ID NO:10.
67 . The antibody or binding fragment of claim 65 which has a 2- to 30-fold greater than the affinity of an antibody having a V H sequence corresponding to SEQ ID NO:1 and a V L sequence corresponding to SEQ ID NO:2.
68 . The antibody or binding fragment of claim 66 which has a 2- to 30-fold greater than the affinity of an antibody having a heavy chain sequence corresponding to SEQ ID NO:9 and a light chain sequence corresponding to SEQ ID NO:10
69 . The antibody or binding fragment of any one of claims 65 to 68 , wherein the affinity is a measure of K D as analyzed by a biosensor.
70 . The antibody or binding fragment of any one of claims 1 to 69 which is purified.
71 . The antibody or binding fragment of claim 70 which is purified to at least 85%, at least 90%, at least 95% or at least 98% homogeneity.
72 . The antibody or binding fragment of any one of claims 1 - 71 , wherein the heavy chain includes at least one substitution selected from A61F in CDR-H2, A61E in CDR-H2, A61D in CDR-H2, D62L in CDR-H2, D62G in CDR-H2, D62Q in CDR-H2, D62T in CDR-H2, D62K in CDR-H2, D62R in CDR-H2, D62E in CDR-H2, D62H in CDR-H2, K64S in CDR-H2, K64V in CDR-H2, K64Q in CDR-H2, R65V in CDR-H2, R65F in CDR-H2, R65H in CDR-H2, R65N in CDR-H2, R65S in CDR-H2, R65Q in CDR-H2, R65K in CDR-H2, R65I in CDR-H2, and Y98H in CDR-H3.
73 . An antibody-drug conjugate comprising an anti-VEGF antibody or anti-VEGF binding fragment according to any one of claims 1 to 72 .
74 . A pharmaceutical composition comprising an anti-VEGF antibody or anti-VEGF binding fragment according to any one of claims 1 to 72 or an antibody-drug conjugate according to claim 73 , and a pharmaceutically acceptable carrier.
75 . A nucleic acid comprising a nucleotide sequence encoding anti-VEGF antibody or anti-VEGF binding fragment of any one of claims 1 to 72 .
76 . A vector comprising the nucleic acid of claim 75 .
77 . A prokaryotic host cell transformed with a vector according to claim 76 .
78 . A eukaryotic host cell transformed with a vector according to claim 76 .
79 . A eukaryotic host cell engineered to express the nucleotide sequence of claim 75 .
80 . The eukaryotic host cell of claim 79 which is a mammalian host cell.
81 . A method of producing anti-VEGF antibody or anti-VEGF binding fragment, comprising: (a) culturing the eukaryotic host cell of claim 79 or claim 80 and (b) recovering the anti-VEGF antibody or anti-VEGF binding fragment antibody.
82 . A method of treating cancer, comprising administering to a human patient in need thereof a therapeutically effective amount of anti-VEGF antibody or anti-VEGF binding fragment according to any one of claims 1 to 72 , an antibody-drug conjugate according to claim 73 , or a pharmaceutical composition according to claim 74 .
83 . The method of claim 82 , wherein the cancer is metastatic carcinoma of the colon, metastatic carcinoma of the rectum, non-squamous non-small cell lung cancer, or metastatic HER2-negative breast cancer.
84 . The method of claim 83 , wherein the cancer is non-squamous non-small cell lung cancer which is unresectable, locally advanced, recurrent or metastatic.
85 . A method of treating age-related macular degeneration, comprising administering to a human patient in need thereof a therapeutically effective amount of anti-VEGF antibody or anti-VEGF binding fragment according to any one of claims 1 to 72 , an antibody-drug conjugate according to claim 73 , or a pharmaceutical composition according to claim 74 .
86 . A method of treating an immune disorder, comprising administering to a human patient in need thereof a therapeutically effective amount of anti-VEGF antibody or anti-VEGF binding fragment according to any one of claims 1 to 72 , an antibody-drug conjugate according to claim 73 , or a pharmaceutical composition according to claim 74 .
87 . The method of claim 86 , wherein the immune disorder is rheumatoid arthritis or Grave's disease.Cited by (0)
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