Compositions and methods for the depletion of CD137+ cells
The invention provides methods of preventing and treating graft-versus-host-disease and autoimmune diseases, such as those arising from transplant therapy, by selective depletion of hematopoietic cells through the use of antibody-drug conjugates and ligand-drug conjugates that specifically bind CD137. The compositions and methods described herein can be used to treat a variety of pathologies, including stem cell disorders and other blood conditions.
1. An antibody drug conjugate (ADC) comprising an anti-CD137 antibody, or antigen binding portion thereof, said antibody comprising a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 having an amino acid sequence as set forth in SEQ ID NOs: 25, 26, and 27, respectively, and comprising a light chain variable region comprising a CDR1, a CDR2, and a CDR3 having an amino acid sequence as set forth in SEQ ID NOs: 29, 30, and 31, respectively, wherein the antibody, or antigen binding portion thereof, is conjugated at a cysteine residue to an amatoxin via a linker, and wherein the amatoxin is represented by formula (IV)
wherein R 1 is H, OH, or OR A ;
R 2 is H, OH, or OR B ;
R A and R B , together with the oxygen atoms to which they are bound, combine to form a 5-membered heterocyclolalkyl group;
R 3 is H or R D ;
R 4 , R 6 , and R 7 are each independently H, OH, OR D , or R D ;
R 5 is H, OH, OR C , OR D , R C , or R D ;
R 8 is OH, NH 2 , OR C , OR D , NHR C , or NR C R D ;
R 9 is H, OH, or OR D ;
X is —S—, —S(O)—, or —SO 2 —;
R C is -L-Z;
R D is C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 2 -C 6 alkenyl, C 2 -C 6 heteroalkenyl, C 2 -C 6 alkynyl, C 2 -C 6 heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;
L is C 1 -C 6 alkylene, C 1 -C 6 heteroalkylene, C 2 -C 6 alkenylene, C 2 -C 6 heteroalkenylene, C 2 -C 6 alkynylene, C 2 -C 6 heteroalkynylene, cycloalkylene, heterocycloalkylene, arylene, heteroarylene, or comprises a dipeptide; and
Z is a chemical moiety formed from a coupling reaction between a reactive substituent present on L and a reactive substituent present within the antibody or antigen-binding fragment thereof,
wherein the amatoxin comprises exactly one R C substituent.
2. The ADC of claim 1 , wherein the amatoxin is an amanitin.
3. The ADC of claim 2 , wherein the amanitin is selected from the group consisting of α-amanitin, β-amanitin, γ-amanitin, ε-amanitin, amanin, amaninamide, amanullin, amanullinic acid, and proamanullin.
4. A pharmaceutical composition comprising the ADC of claim 1 , and a pharmaceutically acceptable carrier.
5. The ADC of claim 1 , wherein the heavy chain variable region comprises the amino acid sequence as set forth in SEQ ID NO: 28 and the light chain variable region comprises the amino acid sequence as set forth in SEQ ID NO: 32.
6. The ADC of claim 1 , wherein the antibody is an IgG1 or an IgG4 isotype.
7. The ADC of claim 1 , wherein the antibody is BBK2.
8. The ADC of claim 1 , wherein the amatoxin is α-amanitin.
9. The ADC of claim 1 , wherein R 5 is OR C .
10. The ADC of claim 1 , wherein R 5 is R C .
11. The ADC of claim 1 , wherein R 8 is OR C .
12. The ADC of claim 1 , wherein R 8 is NHR C .
13. The ADC of claim 1 , wherein R 8 is NR C R D .
14. The ADC of claim 1 , wherein R 1 and R 2 are each OH.
15. The ADC of claim 1 , wherein L is C 1 -C 6 alkylene, C 1 -C 6 heteroalkylene, C 2 -C 6 alkenylene, C 2 -C 6 heteroalkenylene, C 2 -C 6 alkynylene, C 2 -C 6 heteroalkynylene, or comprises a dipeptide.
16. The ADC of claim 15 , wherein L is C 1 -C 6 alkylene, C 1 -C 6 heteroalkylene, C 2 -C 6 alkenylene, C 2 -C 6 heteroalkenylene, C 2 -C 6 alkynylene, or C 2 -C 6 heteroalkynylene.
17. The ADC of claim 15 , wherein L comprises a dipeptide.
18. The ADC of claim 1 , wherein X is —S—.
19. The ADC of claim 1 , wherein X is —S(O)—.
20. The ADC of claim 1 , wherein X is —SO 2 —.
21. An antibody drug conjugate (ADC) comprising an anti-CD137 antibody, or antigen binding portion thereof, said antibody comprising a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 having an amino acid sequence as set forth in SEQ ID NOs: 25, 26, and 27, respectively, and comprising a light chain variable region comprising a CDR1, a CDR2, and a CDR3 having an amino acid sequence as set forth in SEQ ID NOs: 29, 30, and 31, respectively, wherein the antibody, or antigen binding portion thereof, is conjugated at a cysteine residue to an amatoxin via a linker, and wherein the amatoxin is represented by formula (IV)
wherein R 1 is H, OH, or OR A ;
R 2 is H, OH, or OR B ;
R A and R B , together with the oxygen atoms to which they are bound, combine to form a 5-membered heterocyclolalkyl group;
R 3 is H or R D ;
R 4 , R 6 , and R 7 are each independently H, OH, OR D , or R D ;
R 5 is OR C or R C ;
R 8 is OH, NH 2 , or OR D ;
R 9 is H, OH, or OR D ;
X is —S—, —S(O)—, or —SO 2 —;
R C is -L-Z;
R D is C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 2 -C 6 alkenyl, C 2 -C 6 heteroalkenyl, C 2 -C 6 alkynyl, C 2 -C 6 heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;
L is C 1 -C 6 alkylene, C 1 -C 6 heteroalkylene, C 2 -C 6 alkenylene, C 2 -C 6 heteroalkenylene, C 2 -C 6 alkynylene, C 2 -C 6 heteroalkynylene, or comprises a dipeptide; and
Z is a chemical moiety formed from a coupling reaction between a reactive substituent present on L and a reactive substituent present within the antibody or antigen-binding fragment thereof.
22. A pharmaceutical composition comprising the ADC of claim 21 , and a pharmaceutically acceptable carrier.
23. An antibody drug conjugate (ADC) comprising an anti-CD137 antibody, or antigen binding portion thereof, said antibody comprising a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 having an amino acid sequence as set forth in SEQ ID NOs: 25, 26, and 27, respectively, and comprising a light chain variable region comprising a CDR1, a CDR2, and a CDR3 having an amino acid sequence as set forth in SEQ ID NOs: 29, 30, and 31, respectively, wherein the antibody, or antigen binding portion thereof, is conjugated at a cysteine residue to an amatoxin via a linker, and wherein the amatoxin is represented by formula (IV)
wherein R 1 is H, OH, or OR A ;
R 2 is H, OH, or OR B ;
R A and R B , together with the oxygen atoms to which they are bound, combine to form a 5-membered heterocyclolalkyl group;
R 3 is H or R D ;
R 4 , R 6 , and R 7 are each independently H, OH, OR D , or R D ;
R 5 is H, OH, OR D , or R D ;
R 8 is OR C , NHR C , or NR C R D ;
R 9 is H, OH, or OR D ;
X is —S—, —S(O)—, or —SO 2 —;
R C is -L-Z;
R D is C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 2 -C 6 alkenyl, C 2 -C 6 heteroalkenyl, C 2 -C 6 alkynyl, C 2 -C 6 heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;
L is C 1 -C 6 alkylene, C 1 -C 6 heteroalkylene, C 2 -C 6 alkenylene, C 2 -C 6 heteroalkenylene, C 2 -C 6 alkynylene, C 2 -C 6 heteroalkynylene, or comprises a dipeptide; and
Z is a chemical moiety formed from a coupling reaction between a reactive substituent present on L and a reactive substituent present within the antibody or antigen-binding fragment thereof.
24. A pharmaceutical composition comprising the ADC of claim 23 , and a pharmaceutically acceptable carrier.