Anti-CD47 antibodies and uses thereof

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

(1) SEQ ID NOs:177, 46, 47, 178, 112, and 179, respectively;

(2) SEQ ID NOs:51, 52, 53, 117, 118, and 119, respectively;

(3) SEQ ID NOs:54, 55, 56, 120, 121, and 122, respectively;

(4) SEQ ID NOs:57, 58, 59, 123, 124, and 125, respectively;

(5) SEQ ID NOs:60, 61, 62, 126, 127, and 128, respectively;

(6) SEQ ID NOs:180, 181, 182, 129, 130, and 131, respectively;

(7) SEQ ID NOs:72, 73, 74, 138, 139, and 140, respectively;

(8) SEQ ID NOs:78, 79, 80, 144, 145, and 146, respectively;

(9) SEQ ID NOs:81, 82, 83, 147, 148, and 149, respectively;

(10) SEQ ID NOs:84, 85, 86, 150, 151, and 152, respectively;

(11) SEQ ID NOs:87, 88, 89, 153, 154, and 155, respectively;

(12) SEQ ID NOs:90, 91, 92, 156, 157, and 158, respectively;

(13) SEQ ID NOs:93, 94, 95, 159, 160, and 161, respectively;

(14) SEQ ID NOs:96, 97, 98, 162, 163, and 164, respectively;

(15) SEQ ID NOs:99, 100, 101, 165, 166, and 167, respectively;

(16) SEQ ID NOs:102, 103, 104, 168, 169, and 170, respectively;

(17) SEQ ID NOs:105, 106, 107, 171, 172, and 173, respectively;

(18) SEQ ID NOs:108, 109, 110, 174, 175, and 176, respectively; or

(19) SEQ ID NOs:201, 202, 203, 204, 205, and 206, respectively;

wherein the antibody or antigen-binding fragment thereof specifically binds CD47.

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

a. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:1, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:2;

b. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:3, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:4;

c. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:5, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:6;

d. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:7, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:8;

e. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:9, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:10;

f. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:11, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:12;

g. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:13, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:14;

h. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:15, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:16;

i. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:17, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:18;

j. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:19, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:20;

k. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:21, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:22;

l. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:23, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:24;

m. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:25, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:26;

n. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:27, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:28;

o. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:29, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:30;

p. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:31, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:32;

q. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:33, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:34;

r. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:35, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:36;

s. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:37, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:38;

t. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:39, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:40;

u. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:41, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:42; or

v. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:43, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:44;

w. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:183, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:191;

x. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:183, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:192;

y. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:183, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:193;

z. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:184, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:190;

aa. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:184, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:192;

bb. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:184, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:193;

cc. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:185, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:190;

dd. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:185, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:191;

ee. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:185, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:193;

ff. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:185, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:198;

gg. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:187, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:194;

hh. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:188, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:194;

ii. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:188, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:196;

jj. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:188, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:197; or

kk. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:199, and a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO:200.

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

b. 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;

c. 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;

d. 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;

e. 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;

f. 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;

g. 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;

h. 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;

i. 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;

j. 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;

k. 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;

l. 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;

m. 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;

n. 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;

o. 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;

p. 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;

q. 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;

r. 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;

s. 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;

t. 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;

u. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:41, and a light chain variable region having the polypeptide sequence of SEQ ID NO:42; or

v. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:43, and a light chain variable region having the polypeptide sequence of SEQ ID NO:44;

w. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:183, and a light chain variable region having the polypeptide sequence of SEQ ID NO:191;

x. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:183, and a light chain variable region having the polypeptide sequence of SEQ ID NO:192;

y. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:183, and a light chain variable region having the polypeptide sequence of SEQ ID NO:193;

z. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:184, and a light chain variable region having the polypeptide sequence of SEQ ID NO:190;

aa. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:184, and a light chain variable region having the polypeptide sequence of SEQ ID NO:192;

bb. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:184, and a light chain variable region having the polypeptide sequence of SEQ ID NO:193;

cc. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:185, and a light chain variable region having the polypeptide sequence of SEQ ID NO:190;

dd. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:185, and a light chain variable region having the polypeptide sequence of SEQ ID NO:191;

ee. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:185, and a light chain variable region having the polypeptide sequence of SEQ ID NO:193;

ff. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:185, and a light chain variable region having the polypeptide sequence of SEQ ID NO:198;

gg. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:187, and a light chain variable region having the polypeptide sequence of SEQ ID NO:194;

hh. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:188, and a light chain variable region having the polypeptide sequence of SEQ ID NO:194;

ii. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:188, and a light chain variable region having the polypeptide sequence of SEQ ID NO:196;

jj. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:188, and a light chain variable region having the polypeptide sequence of SEQ ID NO:197; or

kk. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:199, and a light chain variable region having the polypeptide sequence of SEQ ID NO:200.

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

5. The isolated monoclonal antibody or antigen-binding fragment of claim 1 , wherein the antibody or antigen-binding fragment thereof is capable of blocking binding of CD47 to signal regulatory protein alpha (SIRPα), inducing macrophage-mediated phagocytosis of cancer cells, or binding cancer cells with minimal to undetectable binding to red blood cells.

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 treating cancer in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of claim 9 .

11. A method of determining a level of CD47 in a subject, the method comprising:

a. obtaining a sample from the subject;

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

c. determining a level of CD47 in the subject.

12. The method of claim 11 , wherein the sample is a tissue sample or a blood sample.

13. The method of claim 12 , wherein the tissue sample is a cancer tissue sample.

14. A method of producing the monoclonal antibody or antigen-binding fragment thereof of claim 1 , comprising culturing a cell comprising a nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof under conditions to produce the monoclonal antibody or antigen-binding fragment thereof, and recovering the antibody or antigen-binding fragment thereof from the cell or culture.

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

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

17. The monoclonal antibody or antigen-binding fragment thereof of claim 1 , wherein the monoclonal antibody or antigen-binding fragment thereof specifically binds human CD47.