Immunoconjugates and methods
Immunoconjugates of the Formula (I) include a linking group for linking an antibody targeting ligand (Ab) to a drug (D). Embodiments of such immunoconjugates are useful for delivering the drug to selected cells or tissues, e.g., for the treatment of cancer. Ab-[S-L 1 -L 2 -L 3 -L 4 -L 5 -L 6 -L 7 -D] n (I)
1. An immunoconjugate having Formula (I),
Ab-[S-L 1 -L 2 -L 3 -L 4 -L 5 -L 6 -L 7 -D] n (I)
wherein:
Ab is an antibody or an antigen-binding fragment;
L 1 is
L 2 is absent,
Z 1 and Z 2 are each individually hydrogen, halogen, NO 2 , —O—(C 1 -C 6 alkyl), or C 1 -C 6 alkyl;
L 3 is —(CH 2 ) n 1 —C(═O)— or —(CH 2 CH 2 O) n 1 —(CH 2 ) n 1 C(═O)—;
each n 1 is independently an integer of 0 to 12;
L 4 is a tetrapeptide residue;
L 5 is absent or —[NH(CH 2 ) n 2 ] n 3 —;
n 2 is an integer of 0 to 6;
n 3 is an integer of 0 to 2;
L 6 is absent or
L 7 is absent,
D is a drug moiety; and
n is an integer from 1 to 10, wherein said antibody or antigen-binding fragment thereof comprises
a heavy chain comprising:
VHCDR 1 comprising an amino acid sequence of SEQ ID NO:1;
VHCDR 2 comprising an amino acid sequence of SEQ ID NO:2; and
VHCDR 3 comprising an amino acid sequence of SEQ ID NO:3; and
a light chain comprising:
VLCDR 1 comprising an amino acid sequence of SEQ ID NO:8;
VLCDR 2 comprising an amino acid sequence of AAS; and
VLCDR 3 comprising an amino acid sequence of SEQ ID NO:10;
wherein the antibody or antigen-binding fragment thereof specifically binds to the extracellular domain of human receptor tyrosine kinase like orphan receptor 1 (ROR1).
2. The immunoconjugate of claim 1 , wherein the antibody or antigen-binding fragment thereof comprises
a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 5; and
a light chain variable region comprising an amino acid sequence of SEQ ID NO: 12.
3. The immunoconjugate of claim 1 , wherein the antibody or antigen-binding fragment thereof comprises
a heavy chain comprising an amino acid sequence of SEQ ID NO: 7; and
a light chain comprising an amino acid sequence of SEQ ID NO: 14.
4. The immunoconjugate of claim 1 , wherein D is a drug moiety of Formula (II) having the structure:
or a pharmaceutically acceptable salt thereof, wherein:
R 1 and R 2 are each individually selected from the group consisting of hydrogen, halogen, —CN, —OR 5 , —NR 5 R 6 , a substituted or an unsubstituted C 1 -C 6 alkyl, a substituted or an unsubstituted C 1 -C 6 haloalkyl, a substituted or an unsubstituted —O—(C 1 -C 6 alkyl), a substituted or an unsubstituted —O—(C 1 -C 6 haloalkyl), —[(CY 2 ) p O(CY 2 ) q ] t CY 3 , or a substituted or an unsubstituted —O—(CR 5 R 6 ) m —O— such that R 1 and R 2 taken together form a ring;
R 3 is hydrogen or a substituted or an unsubstituted C 1 -C 6 alkyl, a substituted or an unsubstituted C 1 -C 6 haloalkyl, or —[(CY 2 ) p O(CY 2 ) q ] t CY 3 ;
R 4 is hydrogen, a substituted or an unsubstituted —(C 1 -C 6 alkyl)-X 2 , a substituted or an unsubstituted —(C 1 -C 6 haloalkyl)-X 2 , a substituted or an unsubstituted —(C 1 -C 6 alkenyl)-X 2 , a substituted or an unsubstituted —(C 1 -C 6 haloalkenyl)-X 2 , a substituted or an unsubstituted —(C 1 -C 6 alkynyl)-X 2 , or a substituted or an unsubstituted —(C 1 -C 6 haloalkynyl)-X 2 ;
X 1 is —O—, —S(O n6 )—, —NH—, —O—(C═O)—, —NH—(C═O)—, —NH—(C═O)—O—, —NH—(C═O)—NH—, or —NH—S(O n6 )—;
X 2 is —OR 9 , —SR 9 , or —NHR 9 ;
R 5 and R 6 are each individually hydrogen, halogen, a substituted or an unsubstituted C 1 -C 6 alkyl, a substituted or an unsubstituted C 1 -C 6 haloalkyl, or —[(CY 2 ) p O(CY 2 ) q ] t CY 3 ;
m is 1 or 2;
n 4 and n 5 are each individually 0, 1 or 2, with the proviso that n 4 and n 5 are not both 0;
n 6 is 0, 1 or 2;
each Y is individually H or halogen;
each p is individually 1, 2, 3, 4, 5, or 6;
each q is individually 0, 1, 2, 3, 4, 5, or 6;
each t is individually 1, 2, 3, 4, 5, or 6;
R 7 is H, —COR 8 , —CO 2 R 8 , —(CO)—NHR 8 , L 4 , L 5 , L 6 , or L 7 ;
R 8 is a substituted or an unsubstituted C 1 -C 6 alkyl-X 3 , a substituted or an unsubstituted C 1 -C 6 haloalkyl-X 3 , or —[(CY 2 ) p O(CY 2 ) q ] t CY 2 —X 3 ;
R 9 is H, —COR 8 , —CO 2 R 8 , —(CO)—NHR 8 , L 4 , L 5 , L 6 , or L 7 , with the proviso that exactly one of R 7 and R 9 is L 4 , L 5 , L 6 , or L 7 ; and
each X 3 is individually —H, —OH, —SH, or —NH 2 .
5. The immunoconjugate of claim 4 , wherein:
R 1 and R 2 are each individually selected from the group consisting of hydrogen, halogen, —OR 5 , an unsubstituted C 1 -C 6 alkyl, an unsubstituted C 1 -C 6 haloalkyl, an unsubstituted —O—(C 1 -C 6 alkyl), and an unsubstituted —O—(CR 5 R 6 ) m —O-such that R 1 and R 2 taken together form a ring;
R 3 is hydrogen or an unsubstituted C 1 -C 6 alkyl;
R 4 is —(C 1 -C 6 alkyl)-X 2 ;
X 1 is —O—;
X 2 is —OR 9 ;
R 5 and R 6 are each individually hydrogen;
mis 1;
n 4 is 1 or 2;
n 5 is 0;
R 7 is H; and
R 9 is L 4 , L 5 , L 6 , or L 7 .
6. The immunoconjugate of claim 5 , wherein n 4 is 1.
7. The immunoconjugate of claim 5 , wherein n 4 is 2.
8. The immunoconjugate of claim 5 , wherein:
L 2 is absent;
n 1 5;
L 4 is gly-gly-phe-gly (SEO ID NO: 43);
L 5 is —[NH(CH 2 ) n 2 ] n 3 —;
n 2 is 1;
n 3 is 1; and
L 7 is absent.
9. The immunoconjugate of claim 5 , wherein:
L 2 is
Z 1 and Z 2 are each individually hydrogen;
n 1 is 2;
L 4 is gly-gly-phe-gly (SEO ID NO: 43);
L 5 is —[NH(CH 2 ) n 2 ] n 3 —;
n 2 is 1;
n 3 is 1; and
L 7 is absent.
10. The immunoconjugate of claim 4 , wherein D is a compound of Formula (II) having the structure:
or a pharmaceutically acceptable salt thereof, wherein:
R 1 is methyl;
R 2 is fluoro;
R 3 is hydrogen;
R 4 is an unsubstituted —(C 2 alkyl)-X 2 ; and
n 4 is 2.
11. The immunoconjugate of claim 4 , wherein D is a compound of Formula (II) having the structure:
or a pharmaceutically acceptable salt thereof, wherein R 9 is L 4 , L 5 , L 6 , or L 7 .
12. The immunoconjugate of claim 4 , wherein D is a compound of Formula (II) having the structure:
or a pharmaceutically acceptable salt thereof, wherein R 9 is L 4 , L 5 , L 6 , or L 7 .
13. The immunoconjugate of claim 4 , wherein D is a compound of Formula (II) having the structure:
or a pharmaceutically acceptable salt thereof, wherein R 9 is L 4 , L 5 , L 6 , or L 7 .
14. An immunoconjugate selected from the group consisting of:
wherein:
Ab is an antibody or antigen-binding fragment thereof comprising:
a heavy chain comprising:
VHCDR 1 comprising an amino acid sequence of SEQ ID NO:1;
VHCDR 2 comprising an amino acid sequence of SEQ ID NO:2; and
VHCDR 3 comprising an amino acid sequence of SEQ ID NO:3; and
a light chain comprising:
VLCDR 1 comprising an amino acid sequence of SEQ ID NO:8;
VLCDR 2 comprising an amino acid sequence of AAS; and
VLCDR 3 comprising an amino acid sequence of SEQ ID NO:10;
wherein the antibody or antigen-binding fragment thereof specifically binds to the extracellular domain of human receptor tyrosine kinase like orphan receptor 1 (ROR1).
15. The immunoconjugate of claim 14 , wherein the antibody or antigen-binding fragment thereof comprises
a heavy chain variable region comprising an amino acid sequence identical to SEQ ID NO: 5; and
a light chain variable region comprising an amino acid sequence identical to SEQ ID NO: 12.
16. The immunoconjugate of claim 14 , wherein the antibody or antigen-binding fragment thereof comprises
a heavy chain comprising an amino acid sequence identical to SEQ ID NO: 7; and
a light chain comprising an amino acid sequence identical to SEQ ID NO: 14.
17. The immunoconjugate of claim 14 , wherein the immunoconjugate is:
18. The immunoconjugate of claim 14 , wherein the immunoconjugate is:
19. The immunoconjugate of claim 15 , wherein the immunoconjugate is:
20. The immunoconjugate of claim 15 , wherein the immunoconjugate is:
21. An immunoconjugate selected from the group consisting of:
wherein:
Ab is an antibody or antigen-binding fragment thereof comprising:
a heavy chain comprising:
VHCDR 1 comprising an amino acid sequence of SEQ ID NO:1;
VHCDR 2 comprising an amino acid sequence of SEQ ID NO:2; and
VHCDR 3 comprising an amino acid sequence of SEQ ID NO:3; and
a light chain comprising:
VLCDR 1 comprising an amino acid sequence of SEQ ID NO:8;
VLCDR 2 comprising an amino acid sequence of AAS; and
VLCDR 3 comprising an amino acid sequence of SEQ ID NO:10;
wherein the antibody or antigen-binding fragment thereof specifically binds to the extracellular domain of human receptor tyrosine kinase like orphan receptor 1 (ROR1).
22. The immunoconjugate of claim 21 , wherein the antibody or antigen-binding fragment thereof comprises
a heavy chain variable region comprising an amino acid sequence identical to SEQ ID NO: 5; and
a light chain variable region comprising an amino acid sequence identical to SEQ ID NO: 12.
23. The immunoconjugate of claim 21 , wherein the antibody or antigen-binding fragment thereof comprises
a heavy chain comprising an amino acid sequence identical to SEQ ID NO: 7; and
a light chain comprising an amino acid sequence identical to SEQ ID NO: 14.
24. The immunoconjugate of claim 21 , wherein the immunoconjugate is:
25. The immunoconjugate of claim 21 , wherein the immunoconjugate is:
26. The immunoconjugate of claim 22 , wherein the immunoconjugate is:
27. The immunoconjugate of claim 22 , wherein the immunoconjugate is:
28. A method for treating a cancer or a tumor comprising administering an effective amount of an immunoconjugate of claim 21 , or a pharmaceutical composition thereof, to a subject having the cancer or the tumor, wherein the cancer or the tumor expresses the extracellular domain of human receptor tyrosine kinase-like orphan receptor (ROR1).
29. The method of claim 28 , wherein the cancer or the tumor is selected from lung cancer, urothelial cancer, colorectal cancer, prostate cancer, ovarian cancer, pancreatic cancer, breast cancer, bladder cancer, gastric cancer, gastrointestinal stromal tumor, uterine cervix cancer, esophageal cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, vulval cancer, thyroid cancer, penis cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma, and sarcoma.
30. A method for treating a cancer or a tumor comprising administering an effective amount of an immunoconjugate of claim 14 , or a pharmaceutical composition thereof, to a subject having the cancer or the tumor, wherein the cancer or the tumor expresses the extracellular domain of human receptor tyrosine kinase-like orphan receptor (ROR1); and the cancer or the tumor is selected from lung cancer, urothelial cancer, colorectal cancer, prostate cancer, ovarian cancer, pancreatic cancer, breast cancer, bladder cancer, gastric cancer, gastrointestinal stromal tumor, uterine cervix cancer, esophageal cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, vulval cancer, thyroid cancer, penis cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma, and sarcoma.