ANTI-FAMILY WITH SEQUENCE SIMILARITY 19, MEMBER A5 ANTIBODIES AND METHOD OF USE THEREOF
The present disclosure provides antibodies that specifically bind to human FAM19A5 and compositions comprising such antibodies. In some embodiments, antibodies are de-immunized to reduce immunogenicity in a human subject. In certain embodiments, antibodies have undergone affinity maturation. In some embodiments, the anti-FAM19A5 antibodies can modulate FAM19A5 activity, e.g, inhibit, suppress, reduce, or reverse the onset of reactive gliosis and/or excessive proliferation of reactive astrocytes, utilizing such antibodies. The present disclosure also provides methods for treating disorders, such as central nervous system damage, a degenerative brain disorder, a neuropathic pain, or a cancer, by administering an antibody that specifically binds to human FAM19A5.
1 . An isolated antibody, or antigen binding portion thereof, which specifically binds a human family with sequence similarity 19, member A5 (FAM19A5) protein (“anti-FAM19A5 antibody”), comprising a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3,
wherein the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequence set forth in SEQ ID NOs: 5, 6, and 7, or with at least one amino acid mutation,
wherein the light chain CDR1, CDR2, and CDR3 comprise the amino acid sequence set forth in SEQ ID NOs: 8, 9, and 10, respectively, wherein at least one of the light chain CDR1, CDR2, and CDR3 comprises at least one amino acid mutation, and
wherein the anti-FAM19A5 antibody has a reduced immunogenicity compared to a reference antibody comprising the heavy chain variable region (VH) set forth in SEQ ID NO: 11 and the light chain variable region (VL) set forth in SEQ ID NO: 12.
2 . The anti-FAM19A5 antibody of claim 1 , wherein: (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 5 or with one or two amino acid mutations; (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 6 with one, two, three, four, or five amino acid mutations; (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 7; (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 8 or with one, two, three, or four amino acid mutations; (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 9 with one, two, three, or four amino acid mutations; and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 10 or with one, two, three, four, or five amino acid mutations.
3 . (canceled)
4 . The anti-FAM19A5 antibody of claim 2 , wherein;
(a) the amino acid mutation of the heavy chain CDR1 comprises (i) a substitution of Threonine at amino acid 3 of SEQ ID NO: 5 to an acidic amino acid, (ii) a substitution of Serine at amino acid 5 of SEQ ID NO: 5 to an acidic amino acid, or (iii) both (i) and (ii);
(b) the amino acid mutation of the heavy chain CDR2 comprises (i) a substitution of Arginine at amino acid 16 of SEQ ID NO: 6 to a basic amino acid, (ii) a substitution of Glycine at amino acid 6 of SEQ ID NO: 6 to an acidic amino acid, (iii) a substitution of Serine at amino acid 7 of SEQ ID NO: 6 to an acidic amino acid, (iv) a substitution of Serine at amino acid 8 of SEQ ID NO: 6 to an acidic amino acid, (v) a substitution of Threonine at amino acid 9 of SEQ ID NO: 6 to an acidic amino acid, (vi) a substitution of Arginine at amino acid 16 of SEQ ID NO: 6 to a basic amino acid, or (vii) a combination thereof;
(c) the amino acid mutation of the light chain CDR1 comprises (i) a substitution of Tyrosine at amino acid 6 of SEQ ID NO: 8 to an acidic amino acid, (ii) a substitution of Arginine at amino acid 7 of SEQ ID NO: 8 to an acidic amino acid, (iii) a substitution of Glycine at amino acid 8 of SEQ ID NO: 8 to an acidic amino acid, (iv) a substitution of Serine at amino acid 9 of SEQ ID NO: 8 to an acidic amino acid, or (v) a combination thereof;
(d) the amino acid mutation of the light chain CDR2 comprises (i) a substitution of Glutamic Acid at amino acid 1 of SEQ ID NO: 9 to an acidic amino acid, (ii) a substitution of Serine at amino acid 2 of SEQ ID NO: 9 to an acidic amino acid, (iii) a substitution of Asparagine at amino acid 3 of SEQ ID NO: 9 to an acidic amino acid, to a basic amino acid, or to an aliphatic amino acid, (iv) a substitution of Lysine at amino acid 4 of SEQ ID NO: 9 to an acidic amino acid or to an aliphatic amino acid, or (v) a combination thereof;
(e) the amino acid mutation of the light chain CDR3 comprises (i) a substitution of Serine at amino acid 6 of SEQ ID NO: 10 to an acidic amino acid or to an aliphatic amino acid, (ii) a substitution of Asparagine at amino acid 7 of SEQ ID NO: 10 to an acidic amino acid or to a hydroxyl or sulfur/selenium-containing amino acid, (iii) a substitution of Glycine at amino acid 8 of SEQ ID NO: 10 to an acidic amino acid or to a hydroxyl or sulfur/selenium-containing amino acid, (iv) a substitution of Glycine at amino acid 9 of SEQ ID NO: 10 to an acidic amino acid or to a hydroxyl or sulfur/selenium-containing amino acid, (v) a substitution of Isoleucine at amino acid 10 of SEQ ID NO: 10 to a basic amino acid, or (vi) a combination thereof; or
(f) any combination of (a) to (e).
5 - 28 . (canceled)
29 . The anti-FAM19A5 antibody of claim 1 , wherein:
(a) (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 5; (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 13; (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 7; (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 8; (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 20; and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 10;
(b) (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 14; (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 15; (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 7; (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 21; (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 22; and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 23;
(c) (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 14; (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 15; (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 7; (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 21; (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 24; and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 23;
(d) (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 14; (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 15; (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 7; (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 8; (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 25; and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 23;
(e) (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 14; (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 15; (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 7; (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 8; (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 24; and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 23;
(f) (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 14; (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 15; (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 7; (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 8; (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 26; and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 27; or
(g) (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 14; (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 15; (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 7; (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 8; (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 28; and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 29.
30 - 35 . (canceled)
36 . The anti-FAM19A5 antibody of claim 1 , comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises an amino acid sequence which is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 11; and/or wherein the VL comprises an amino acid sequence which is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 12.
37 - 39 . (canceled)
40 . An isolated antibody, or antigen binding portion thereof, which specifically binds a human family with sequence similarity 19, member A5 (FAM19A5) protein (“anti-FAM19A5 antibody”), comprising a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3,
wherein the heavy chain CDR1, CDR2, and CDR3 comprise the amino acid sequence set forth in SEQ ID NOs: 16, 17, and 18, or with at least one amino acid mutation,
wherein the light chain CDR1, CDR2, and CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 30, 31, and 32, respectively, wherein at least one of the light chain CDR1, CDR2, and CDR3 comprises at least one amino acid mutation, and
wherein the anti-FAM19A5 antibody has a reduced immunogenicity and/or higher binding affinity to human FAM19A5 protein compared to a reference antibody comprising the heavy chain variable region (VH) set forth in SEQ ID NO: 35 and the light chain variable region (VL) set forth in SEQ ID NO: 45.
41 . The anti-FAM19A5 antibody of claim 40 , wherein: (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 16 or with one amino acid mutation; (ii) the heavy chain CDR2 comprises the amino acid set forth in SEQ ID NO: 17; (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 18 or with one or two amino acid mutations; (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 30 with one amino acid mutation; (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 31; and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 32.
42 . The anti-FAM19A5 antibody of claim 40 , wherein;
(a) (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 16, (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 17, (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 18, (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 80, (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 31, and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 32;
(b) (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 19, (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 17, (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 18, (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 80, (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 31, and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 32;
(c) (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 19, (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 17, (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 128, (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 80, (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 31, and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 32; or
(d) (i) the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 19, (ii) the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 17, (iii) the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 129, (iv) the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 80, (v) the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 31, and (vi) the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 32.
43 - 47 . (canceled)
48 . The anti-FAM19A5 antibody of claim 41 , wherein;
(a) the amino acid mutation of the heavy chain CDR1 comprises a substitution of Threonine at amino acid 3 of SEQ ID NO: 16 to an acidic amino acid; (b) the amino acid mutation of the heavy chain CDR3 comprises (i) a substitution of Threonine at amino acid 2 of SEQ ID NO: 18 to a hydroxyl or sulfur/selenium-containing amino acid or to an acid amino acid, (ii) a substitution of Glutamic Acid at amino acid 4 of SEQ ID NO: 18 to an aliphatic amino acid, or (iii) both (i) and (ii); (c) the amino acid mutation of the light chain CDR1 comprises a substitution of Serine at amino acid 4 of SEQ ID NO: 30 to an aliphatic amino acid; or (d) any combination of (a) to (c).
49 . (canceled)
50 . The anti-FAM19A5 antibody of claim 40 , comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises an amino acid sequence which is at least about 80% identical to the amino acid sequence set forth in SEQ ID NO: 35; and/or wherein the VL comprises an amino acid sequence which is at least about 80%, at least about 85%.
51 - 56 . (canceled)
57 . The anti-FAM19A5 antibody of claim 1 , which comprises a Fab, a Fab′, a F(ab′) 2, a Fv, or a single chain Fv (scFv).
58 . The anti-FAM19A5_antibody of claim 40 , which comprises a Fab′, a F(ab′) 2, a Fv, or a single chain Fc (scFv).
59 - 60 . (canceled)
61 . A nucleic acid encoding the anti-FAM19A5 antibody of claim 1 .
62 . A vector comprising the nucleic acid of claim 61 .
63 . A cell comprising the vector of claim 62 .
64 . An immunoconjugate comprising the anti-FAM19A5 antibody of claim 1 , linked to an agent.
65 . A composition comprising the anti-FAM19A5 antibody of claim 1 , and a carrier.
66 . (canceled)
67 . A method of producing an antibody which specifically binds to a human FAM19A5 protein, comprising culturing the cell of claim 63 under suitable conditions and isolating the antibody.
68 . A method of treating a disease or condition in a subject in need thereof, comprising administering to the subject the anti-FAM19A5 antibody of claim 1 .
69 - 88 . (canceled)
89 . A nucleic acid encoding the anti-FAM19A5 antibody of claim 40 .