Anti-CD73 antibodies and uses thereof

1. An isolated monoclonal antibody or antigen-binding fragment thereof comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3, a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3, having the polypeptide sequences of:

a. SEQ ID NOs:89, 90, 91, 149, 150 and 151, respectively, or SEQ ID NOs:209, 210, 211, 269, 270 and 271, respectively;

b. SEQ ID NOs:41, 42, 43, 101, 102 and 103, respectively, or SEQ ID NOs:161, 162, 163, 221, 222 and 223, respectively;

c. SEQ ID NOs:44, 45, 46, 104, 105 and 106, respectively, or SEQ ID NOs:164, 165, 166, 224, 225 and 226, respectively;

d. SEQ ID NOs:47, 48, 49, 107, 108 and 109, respectively, or SEQ ID NOs:167, 168, 169, 227, 228 and 229, respectively;

e. SEQ ID NOs:50, 51, 52, 110, 111 and 112, respectively, or SEQ ID NOs:170, 171, 172, 230, 231 and 232, respectively;

f. SEQ ID NOs:53, 54, 55, 113, 114 and 115, respectively, or SEQ ID NOs:173, 174, 175, 233, 234 and 235, respectively;

g. SEQ ID NOs:56, 57, 58, 116, 117 and 118, respectively, or SEQ ID NOs:176, 177, 178, 236, 237 and 238, respectively;

h. SEQ ID NOs:59, 60, 61, 119, 120 and 121, respectively, or SEQ ID NOs:179, 180, 181, 239, 240 and 241, respectively;

i. SEQ ID NOs:62, 63, 64, 122, 123 and 124, respectively, or SEQ ID NOs:182, 183, 184, 242, 243 and 244, respectively;

j. SEQ ID NOs:65, 66, 67, 125, 126 and 127, respectively, or SEQ ID NOs:185, 186, 187, 245, 246 and 247, respectively;

k. SEQ ID NOs:68, 69, 70, 128, 129 and 130, respectively, or SEQ ID NOs:188, 189, 190, 248, 249 and 250, respectively;

l. SEQ ID NOs:71, 72, 73, 131, 132 and 133, respectively, or SEQ ID NOs:191, 192, 193, 251, 252 and 253, respectively;

m. SEQ ID NOs:74, 75, 76, 134, 135 and 136, respectively, or SEQ ID NOs:194, 195, 196, 254, 255 and 256, respectively;

n. SEQ ID NOs:77, 78, 79, 137, 138 and 139, respectively, or SEQ ID NOs:197, 198, 199, 257, 258 and 259, respectively;

o. SEQ ID NOs:80, 81, 82, 140, 141 and 142, respectively, or SEQ ID NOs:200, 201, 202, 260, 261 and 262, respectively;

p. SEQ ID NOs:83, 84, 85, 143, 144 and 145, respectively, or SEQ ID NOs:203, 204, 205, 263, 264 and 265, respectively;

q. SEQ ID NOs:86, 87, 88, 146, 147 and 148, respectively, or SEQ ID NOs:206, 207, 208, 266, 267 and 268, respectively;

r. SEQ ID NOs:92, 93, 94, 152, 153 and 154, respectively, or SEQ ID NOs:212, 213, 214, 272, 273 and 274, respectively;

s. SEQ ID NOs:95, 96, 97, 155, 156 and 157, respectively, or SEQ ID NOs:215, 216, 217, 275, 276 and 277, respectively; or

t. SEQ ID NOs:98, 99, 100, 158, 159 and 160, respectively, or SEQ ID NOs:218, 219, 220, 278, 279 and 280, respectively;

wherein the antibody or antigen-binding fragment thereof specifically binds to CD73.

2. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , comprising a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:286 33, 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 35, 37, 39, 282, 283, 284, 285, 287, 288, 289, 295, 296, 297, or 298, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:293 34, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 36, 38, 40, 290, 291, 292, 294, or 299.

3. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , comprising:

a. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:286, and a light chain variable region having the polypeptide sequence of SEQ ID NO:293;

b. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:33, and a light chain variable region having the polypeptide sequence of SEQ ID NO:34;

c. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:1, and a light chain variable region having the polypeptide sequence of SEQ ID NO:2;

d. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:3, and a light chain variable region having the polypeptide sequence of SEQ ID NO:4;

e. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:5, and a light chain variable region having the polypeptide sequence of SEQ ID NO:6;

f. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:7, and a light chain variable region having the polypeptide sequence of SEQ ID NO:8;

g. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:9, and a light chain variable region having the polypeptide sequence of SEQ ID NO:10;

h. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:11, and a light chain variable region having the polypeptide sequence of SEQ ID NO:12;

i. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:13, and a light chain variable region having the polypeptide sequence of SEQ ID NO:14;

j. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:15, and a light chain variable region having the polypeptide sequence of SEQ ID NO:16;

k. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:17, and a light chain variable region having the polypeptide sequence of SEQ ID NO:18;

l. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:19, and a light chain variable region having the polypeptide sequence of SEQ ID NO:20;

m. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:21, and a light chain variable region having the polypeptide sequence of SEQ ID NO:22;

n. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:23, and a light chain variable region having the polypeptide sequence of SEQ ID NO:24;

o. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:25, and a light chain variable region having the polypeptide sequence of SEQ ID NO:26;

p. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:27, and a light chain variable region having the polypeptide sequence of SEQ ID NO:28;

q. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:29, and a light chain variable region having the polypeptide sequence of SEQ ID NO:30;

r. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:31, and a light chain variable region having the polypeptide sequence of SEQ ID NO:32;

s. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:35, and a light chain variable region having the polypeptide sequence of SEQ ID NO:36;

t. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:37, and a light chain variable region having the polypeptide sequence of SEQ ID NO:38;

u. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:39, and a light chain variable region having the polypeptide sequence of SEQ ID NO:40;

v. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:282, and a light chain variable region having the polypeptide sequence of SEQ ID NO:290;

w. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:282, and a light chain variable region having the polypeptide sequence of SEQ ID NO:291;

x. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:282, and a light chain variable region having the polypeptide sequence of SEQ ID NO:292;

y. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:283, and a light chain variable region having the polypeptide sequence of SEQ ID NO:290;

z. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:283, and a light chain variable region having the polypeptide sequence of SEQ ID NO:291;

aa. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:283, and a light chain variable region having the polypeptide sequence of SEQ ID NO:292;

bb. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:284, and a light chain variable region having the polypeptide sequence of SEQ ID NO:290;

cc. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:284, and a light chain variable region having the polypeptide sequence of SEQ ID NO:291,

dd. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:284, and a light chain variable region having the polypeptide sequence of SEQ ID NO:292,

ee. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:285, and a light chain variable region having the polypeptide sequence of SEQ ID NO:290,

ff. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:285, and a light chain variable region having the polypeptide sequence of SEQ ID NO:291,

gg. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:285, and a light chain variable region having the polypeptide sequence of SEQ ID NO:292,

hh. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:283, and a light chain variable region having the polypeptide sequence of SEQ ID NO:293,

ii. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:284, and a light chain variable region having the polypeptide sequence of SEQ ID NO:293,

jj. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:285, and a light chain variable region having the polypeptide sequence of SEQ ID NO:293,

kk. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:286, and a light chain variable region having the polypeptide sequence of SEQ ID NO:290,

ll. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:286, and a light chain variable region having the polypeptide sequence of SEQ ID NO:291,

mm. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:286, and a light chain variable region having the polypeptide sequence of SEQ ID NO:292;

nn. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:287, and a light chain variable region having the polypeptide sequence of SEQ ID NO:294,

oo. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:288, and a light chain variable region having the polypeptide sequence of SEQ ID NO:294,

pp. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:289, and a light chain variable region having the polypeptide sequence of SEQ ID NO:294,

qq. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:284, and a light chain variable region having the polypeptide sequence of SEQ ID NO:299,

rr. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:295, and a light chain variable region having the polypeptide sequence of SEQ ID NO:299,

ss. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:296, and a light chain variable region having the polypeptide sequence of SEQ ID NO:299,

tt. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:297, and a light chain variable region having the polypeptide sequence of SEQ ID NO:299, or

uu. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:298, and a light chain variable region having the polypeptide sequence of SEQ ID NO:299.

4. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , wherein the monoclonal antibody or antigen-binding fragment thereof inhibits the enzyme activity of soluble and/or cell-surface CD73, prevents the dimerization of CD73, induces the internalization of CD73, and/or is capable of activating T cells.

5. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof is chimeric or human or humanized.

6. An isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof of claim 1 .

7. A vector comprising the isolated nucleic acid of claim 6 .

8. A host cell comprising the vector of claim 7 .

9. A pharmaceutical composition, comprising the isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier.

10. A method of inhibiting the nucleotidase activity of CD73, preventing the dimerization of CD73, inducing the internalization of CD73, or treating cancer in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of claim 9 .

11. A method of producing a monoclonal antibody or antigen-binding fragment thereof, comprising culturing the host cell of claim 8 under conditions to produce the monoclonal antibody or antigen-binding fragment thereof, and recovering the antibody or antigen-binding fragment thereof from the host cell.

12. A method of producing a pharmaceutical composition comprising the monoclonal antibody or antigen-binding fragment of claim 1 , comprising combining the monoclonal antibody or antigen-binding fragment with a pharmaceutically acceptable carrier to obtain the pharmaceutical composition.

13. A method of determining a level of CD73 or determining the ecto-5′ nucleotidase activity of CD73 in a subject, the method comprising:

a. obtaining a sample from the subject;

b. contacting the sample with an isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 ; and

c. determining a level of CD73 or determining the ecto-5′ nucleotidase activity of CD73 in the subject.

14. The method of claim 13 , wherein the sample is a tissue sample or a blood sample.

15. The method of claim 14 , wherein the tissue sample is a cancer tissue sample.

16. An isolated bispecific antibody or antigen-binding fragment thereof comprising the monoclonal antibody or antigen-binding fragment thereof of claim 1 .

17. An isolated nucleic acid encoding the bispecific antibody or antigen-binding fragment thereof of claim 16 .

18. A vector comprising the isolated nucleic acid of claim 17 .

19. A host cell comprising the vector of claim 18 .

20. A pharmaceutical composition comprising the isolated bispecific antibody or antigen-binding fragment thereof of claim 16 and a pharmaceutically acceptable carrier.