Anti-CEA Antibody Drug Conjugates and Methods of Use

1 . An antibody drug conjugate of the formula Ab4C-L-(D) m ) n , or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein:

Ab is an antibody or antigen-binding fragment thereof, which binds to human CEA and which comprises:

(i) three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:24,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:25,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:26, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:27,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:28,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:23; or

(ii) three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:7,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:8,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:9, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:10,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:11,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:6; or

(iii) three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:41,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:42,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:43, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:44,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:45,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:40;

C is a conjugator;

L is a linker;

D is a cytotoxic agent:

m is an integer from 1 to 8; and

n is from 1 to 10.

2 . (canceled)

3 . The antibody drug conjugate of claim 1 , wherein m is 1.

4 . The antibody drug conjugate of claim 1 , wherein n is from 3 to 10.

5 . The antibody drug conjugate of claim 4 , wherein n is about 8.

6 . The antibody drug conjugate of claim 1 , wherein C is a formula selected from (C-I), (C-Ia), (C-Ib), (C-II), (C-III), (C-IIIa), or (C-IV):

and

* marks the bond where C connects to Ab.

7 . The antibody drug conjugate of claim 6 , wherein C is

8 . The antibody drug conjugate of claim 1 , wherein

L is a formula selected from (L-I), (L-II), or (L-III):

wherein Su is a hydrophilic residue; and

* marks the bond where L connects to C.

9 . The antibody drug conjugate of claim 8 , wherein Su is

10 . The antibody drug conjugate of claim 9 , wherein Su is

11 . The antibody drug conjugate of claim 1 , wherein L is

wherein * marks the bond where L connects to C.

12 . The antibody drug conjugate of claim 1 , wherein the cytotoxic agent is a topoisomerase inhibitor.

13 . The antibody drug conjugate of claim 1 , wherein D is:

wherein

Y is -A-B-C′-D′-*, wherein * marks the bond where D connects to L;

A is a bond, CR 1 R 2 , or N-R 1 ;

B is a bond, —C(═O)—, or —C(═O)O—;

C′ is a bond or a divalent group, wherein the divalent group is unsubstituted or substituted C 1-8 alkyl, unsubstituted or substituted cycloalkyl, unsubstituted or substituted heterocyclyl, unsubstituted or substituted aryl, or unsubstituted or substituted heteroaryl;

D 1 is a bond, NH, or O;

each of R 1 and R 2 is, independently, hydrogen, halogen, substituted or unsubstituted alkyl, or substituted or unsubstituted alkoxyl; or R 1 and R 2 together with the atom to which they are attached, form unsubstituted or substituted cycloalkyl, unsubstituted or substituted heterocyclyl, unsubstituted or substituted aryl, or unsubstituted or substituted heteroaryl;

each of R 3 and R 4 is, independently, hydrogen, halogen, substituted or unsubstituted alkyl, or substituted or unsubstituted alkoxyl; or R 3 and R 4 together with the atoms to which they are attached, form unsubstituted or substituted cycloalkyl, unsubstituted or substituted heterocyclyl, unsubstituted or substituted aryl, or unsubstituted or substituted heteroaryl.

14 . The antibody drug conjugate of claim 1 , wherein D is:

wherein R 7 and R 8 are each independently hydrogen, halogen, or alkyl.

15 . The antibody drug conjugate of claim 1 , wherein D is selected from:

16 . The antibody drug conjugate of claim 15 , wherein D is

17 . The antibody drug conjugate of claim 1 , wherein C-L-(D) m is:

18 . The antibody drug conjugate of claim 1 , wherein C-L-(D) m is:

19 . The antibody drug conjugate of claim 1 , or a tautomer, pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein the antibody drug conjugate has one of the following formulas:

20 . The antibody drug conjugate of claim 1 , wherein the antibody or antigen-binding fragment comprises:

(i) a heavy chain variable region comprising SEQ ID NO. 31, and a light chain variable region comprising SEQ ID NO:32;

(ii) a heavy chain variable region comprising SEQ ID NO:48, and a light chain variable region comprising SEQ ID NO:49; or

(iii) a heavy chain variable region comprising SEQ ID NO:14, and a light chain variable region comprising SEQ ID NO:15.

21 - 24 . (canceled)

25 . The antibody drug conjugate of claim 1 , wherein the antibody or antigen-binding fragment thereof comprises a heavy chain constant region of the subclass of IgG1, and a light chain constant region of the type of kappa.

26 . An antibody drug conjugate of the formula:

or a tautomer, pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein:

n is from 4 to 10; and

Ab is an antibody or antigen-binding fragment thereof that binds CEA and comprises:

(i) three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:7,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:8,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:9, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:10,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:11,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:6; or

(ii) three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:24,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:25,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:26, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:27,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:28,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:23; or

(iii) three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:41,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:42,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:43, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:44,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:45,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:40.

27 . An antibody drug conjugate, or a tautomer, pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein

the antibody drug conjugate has the following formula:

n is from 4 to 10; and

Ab is an antibody or antigen-binding fragment thereof that binds CEA and comprises:

(i) three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:7,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:8,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:9, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:10,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:11,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:6; or

(ii) three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:24,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:25,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:26, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:27,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:28,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:23; or

(iii) three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:41,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:42,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:43, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:44,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:45,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:40.

28 . A pharmaceutical composition comprising the antibody drug conjugate of claim 27 , or a tautomer, pharmaceutically acceptable salt, solvate, or hydrate thereof and a pharmaceutically acceptable carrier.

29 . A method of treating a CEA-expressing cancer comprising administering to a subject in need thereof an effective amount of the antibody drug conjugate of claim 1 , or a tautomer, pharmaceutically acceptable salt, solvate, or hydrate thereof.

30 - 33 . (canceled)

34 . A method of producing the antibody drug conjugate of claim 1 , comprising:

(i) culturing a host cell that has been transformed by an isolated nucleic acid comprising a sequence encoding the antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:99 and a light chain comprising an amino acid sequence of SEQ ID NO:100;

(ii) expressing the antibody or antigen-binding fragment thereof;

(iii) recovering the expressed antibody or antigen-binding fragment thereof; and

(iv) conjugating the cytotoxic agent to the antibody or fragment thereof using a linker, such that the antibody drug conjugate is formed.

35 . (canceled)

36 . A pharmaceutical composition comprising the antibody drug conjugate of claim 1 , or a tautomer, pharmaceutically acceptable salt, solvate, or hydrate thereof, and a pharmaceutically acceptable carrier.

37 . The antibody drug conjugate of claim 1 , wherein the antibody drug conjugate has the following formula:

or a tautomer, pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein n is from 4 to 10.

38 . The antibody drug conjugate of claim 37 , wherein Ab comprises:

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:24,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:25,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:26, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:27,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:28,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:23.

39 . The antibody drug conjugate of claim 37 , wherein Ab comprises:

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:41,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:42,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:43, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:44,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:45,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:40.

40 . The antibody drug conjugate of claim 37 , wherein Ab comprises:

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:7,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:8,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:9, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:10,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 11,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:6.

41 . The antibody drug conjugate of claim 37 , wherein the antibody or antigen-binding fragment comprises:

(i) a heavy chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:31, and a light chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:32;

(ii) a heavy chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:48, and a light chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:49; or

(iii) a heavy chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:14, and a light chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:15.

42 . The antibody drug conjugate of claim 37 , wherein the antibody or antigen-binding fragment comprises a heavy chain having an amino acid sequence of SEQ ID NO:99 and a light chain having an amino acid sequence of SEQ ID NO:100.

43 . The antibody drug conjugate of claim 1 , wherein the antibody drug conjugate has the following formula:

or a tautomer, pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein n is from 4 to 10.

44 . The antibody drug conjugate of claim 43 , wherein Ab comprises:

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:24,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:25,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:26, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:27,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:28,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:23.

45 . The antibody drug conjugate of claim 43 , wherein Ab comprises:

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:41,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:42,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:43, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:44,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:45,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:40.

46 . The antibody drug conjugate of claim 43 , wherein Ab comprises:

three heavy chain CDRs:

HCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:7,

HCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:8,

HCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:9, and

three light chain CDRs:

LCDR1 comprising an amino acid sequence as set forth in SEQ ID NO:10,

LCDR2 comprising an amino acid sequence as set forth in SEQ ID NO:11,

LCDR3 comprising an amino acid sequence as set forth in SEQ ID NO:6.

47 . The antibody drug conjugate of claim 43 , wherein the antibody or antigen-binding fragment comprises:

(i) a heavy chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:31, and a light chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:32;

(ii) a heavy chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:48, and a light chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:49; or

(iii) a heavy chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:14, and a light chain variable region comprising a sequence having at least 90% identity to SEQ ID NO:15.

48 . The antibody drug conjugate of claim 43 , wherein the antibody or antigen-binding fragment comprises a heavy chain having an amino acid sequence of SEQ ID NO:99 and a light chain having an amino acid sequence of SEQ ID NO:100.