Granted Patent B2

US 12,251,448 · App. 17/244,721 · Granted Mar 18, 2025

Inventors: Anthony Boitano (Newton, MA); Michael Cooke (Boston, MA); Rahul Palchaudhuri (Somerville, MA); Sean McDonough (Littleton, MA)

Assignee: Heidelberg Pharma Research GmbH

A61K47/6803A61K38/12A61K47/6849C07K16/2896A61K2039/505C07K2317/92

Abstract

Claims (113)

1. A method of depleting a population of CD5+ cells in a human patient, the method comprising:

administering to the human patient an effective amount of an anti-CD5 antibody 5D7 or antigen-binding fragment thereof,

wherein the antibody, or antigen-binding fragment thereof, is conjugated to a cytotoxin, to form an anti-CD5 antibody 5D7 antibody drug conjugate,

wherein the cytotoxin is an amatoxin,

wherein the anti-CD5 antibody 5D7 antibody drug conjugate has a drug to antibody ratio of about 6 or 2, wherein the conjugation is site-specific via a D265C mutation, and

wherein the antibody or antigen-binding fragment thereof conjugated to a cytotoxin is represented by the formula Ab-Z-L-Am, wherein Ab is the antibody or antigen-binding fragment thereof, L is a linker, Z is a chemical moiety, and Am is an amatoxin represented by formula (I)

wherein R 1 is H, OH, OR A , or OR C ;

R 2 is H, OH, OR B , or OR C ;

R A and R B , when present, together with the oxygen atoms to which they are bound, combine to form an 5-membered heterocycloalkyl group;

R 3 is H, R C , or R D ;

R 4 , R 5 , R 6 , and R 7 are each independently 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 C , 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; a dipeptide, —C(═O)—, a peptide, or a combination thereof; 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 Am comprises exactly one R C substituent, and wherein said antibody or antigen-binding fragment thereof comprises the following complementarity determining regions (CDRs):

a. a CDR-H1 having the amino acid sequence GYTFTNY (SEQ ID NO: 3);

b. a CDR-H2 having the amino acid sequence NTHTGE (SEQ ID NO: 4);

c. a CDR-H3 having the amino acid sequence RGYDWYFDV (SEQ ID NO: 5);

d. a CDR-L1 having the amino acid sequence RASQDINSYLS (SEQ ID NO: 6);

e. a CDR-L2 having the amino acid sequence RANRLVD (SEQ ID NO: 7); and

f. a CDR-L3 having the amino acid sequence QQYDESPWT (SEQ ID NO: 8); or

a. a CDR-H1 having the amino acid sequence FSLSTSGMG (SEQ ID NO: 29);

b. a CDR-H2 having the amino acid sequence WWDDD (SEQ ID NO: 30);

c. a CDR-H3 having the amino acid sequence RRATGTGFDY (SEQ ID NO: 31);

d. a CDR-L1 having the amino acid sequence QDVGTA (SEQ ID NO: 32);

e. a CDR-L2 having the amino acid sequence WTSTRHT (SEQ ID NO: 33);

f. a CDR-L3 having the amino acid sequence YNSYNT (SEQ ID NO: 34).

2. A method of preventing rejection of a hematopoietic stem cell graft in a human patient in need of a hematopoietic stem cell transplant, the method comprising:

administering to the human patient an effective amount of an anti-CD5 antibody 5D7, or antigen-binding fragment thereof, prior to the human patient receiving a transplant comprising hematopoietic stem cells,

wherein the antibody, or antigen-binding fragment thereof, is conjugated to a cytotoxin, to form an anti-CD5 antibody 5D7 antibody drug conjugate,

wherein the cytotoxin is an amatoxin, and

wherein the anti-CD5 antibody 5D7 antibody drug conjugate has a drug to antibody ratio of about 6 or 2, wherein the conjugation is site-specific via a D265C mutation, and

wherein the antibody or antigen-binding fragment thereof conjugated to a cytotoxin is represented by the formula Ab-Z-L-Am, wherein Ab is the antibody or antigen-binding fragment thereof, L is a linker, Z is a chemical moiety, and Am is an amatoxin represented by formula (I)

wherein R 1 is H, OH, OR A , or OR C ;

R 2 is H, OH, OR B , or OR C ;

R A and R B , when present, together with the oxygen atoms to which they are bound, combine to form an 5-membered heterocycloalkyl group;

R 3 is H, R C , or R D ;

R 4 , R 5 , R 6 , and R 7 are each independently 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 C , 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; a dipeptide, —C(═O)—, a peptide, or a combination thereof; 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 Am comprises exactly one R C substituent, and wherein said antibody or antigen-binding fragment thereof comprises the following complementarity determining regions (CDRs):

a. a CDR-H1 having the amino acid sequence GYTFTNY (SEQ ID NO: 3);

b. a CDR-H2 having the amino acid sequence NTHTGE (SEQ ID NO: 4);

c. a CDR-H3 having the amino acid sequence RGYDWYFDV (SEQ ID NO: 5);

d. a CDR-L1 having the amino acid sequence RASQDINSYLS (SEQ ID NO: 6);

e. a CDR-L2 having the amino acid sequence RANRLVD (SEQ ID NO: 7); and

f. a CDR-L3 having the amino acid sequence QQYDESPWT (SEQ ID NO: 8); or

a. a CDR-H1 having the amino acid sequence FSLSTSGMG (SEQ ID NO: 29);

b. a CDR-H2 having the amino acid sequence WWDDD (SEQ ID NO: 30);

c. a CDR-H3 having the amino acid sequence RRATGTGFDY (SEQ ID NO: 31);

d. a CDR-L1 having the amino acid sequence QDVGTA (SEQ ID NO: 32);

e. a CDR-L2 having the amino acid sequence WTSTRHT (SEQ ID NO: 33);

f. a CDR-L3 having the amino acid sequence YNSYNT (SEQ ID NO: 34).

3. The method of claim 1 , comprising administering a transplant comprising hematopoietic stem cells to the human patient.

4. The method of claim 1 , wherein the antibody or antigen-binding fragment thereof,

comprises a V L having the amino acid sequence DIQMTQSPSSMSASLGDRVTITCRASQDINSYLSWFQQKPGKSPKTLIYRANRL VDGVPSRFSGSGSGTDYTLTISSLQYEDFGIYYCQQYDESPWTFGGGTKLEIK (SEQ ID NO: 1); and a V H having the amino acid sequence QIQLVQSGPGLKKPGGSVRISCAASGYTFTNYGMNWVKQAPGKGLRWMGWI NTHTGEPTYADDFKGRFTFSLDTSKSTAYLQINSLRAEDTATYFCTRRGYDWY FDVWGQGTTVTVSS (SEQ ID NO: 2); or

comprises a V L having the amino acid sequence NIVMTQSPSSLSASVGDRVTITCQASQDVGTAVAWYQQKPDQSPKLLIYWTSTRHTGVP DRFTGSGS GTDFTLTISSLQPEDIATYFCHQYNSYNTFGSGTKLEIK (SEQ ID NO: 258); and a V H having the amino acid sequence QVTLKESGPVLVKPTETLTLTCTFSGFSLSTSGMGVGWIRQAPGKGLEWVAHIWWDDD VYYNPSLKSRLTITKDASKDQVSLKLSSVTAADTAVYYCVRRRATGTGFDYWGQGTLV TVSS (SEQ ID NO: 257).

5. The method of claim 1 , wherein R A and R B , together with the oxygen atoms to which they are bound, combine to form a 5 membered heterocycloalkyl group of formula:

wherein Y is —C(═O)—, —C(═S)—, —C(═NR E )—, or —C(R E R E′ )—; and

R E and R E′ are each independently C 1 -C 6 alkylene-R C , C 1 -C 6 heteroalkylene-R C , C 2 -C 6 alkenylene-R C , C 2 -C 6 heteroalkenylene-R C , C 2 -C 6 alkynylene-R C , C 2 -C 6 heteroalkynylene-R C , cycloalkylene-R C , heterocycloalkylene-R C , arylene-R C , or heteroarylene-R C .

6. The method of claim 1 , wherein:

(a) 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:

R 3 , R 4 , R 6 , and R 7 are each H;

R 5 is OR C ;

R 8 is OH or NH 2 ; and

R 9 is H or OH;

(b) Rand R 2 are each independently H or OH;

R 3 is R C ;

R 4 , R 6 , and R 7 are each H;

R 5 is H, OH, or OC 1 -C 6 alkyl;

R 8 is OH or NH 2 ; and

R 9 is H or OH;

(c) R 1 and R 2 are each independently H or OH;

R 3 , R 6 , and R 7 are each H;

R 4 and R 5 are each independently H, OH, OR C , or R C ;

R 8 is OH or NH 2 ; and

R 9 is H or OH;

or

(d) R 1 and R 2 are each independently H or OH;

R 3 , R 6 , and R 7 are each H;

R 4 and R 5 are each independently H or OH;

R 8 is OR C or NHR C ; and

R 9 is H or OH.

7. The method of claim 1 , wherein the antibody or antigen-binding fragment thereof conjugated to a cytotoxin is represented by the formula Ab-L-Z-Am, wherein Ab is the antibody or antigen-binding fragment thereof, Z is a chemical moiety, L is a linker, and Am is an amatoxin, and the amatoxin-linker conjugate Am-L-Z is represented by formula (IIB)

wherein X is S, SO, or SO 2 ;

R 1 is H or a linker covalently bound to the antibody or antigen-binding fragment thereof through a chemical moiety Z, formed from a coupling reaction between a reactive substituent present on the linker and a reactive substituent present within an antibody, or antigen-binding fragment thereof; and

R 2 is H or a linker covalently bound to the antibody or antigen-binding fragment thereof through a chemical moiety Z, formed from a coupling reaction between a reactive substituent present on the linker and a reactive substituent present within an antibody, or antigen-binding fragment thereof;

wherein when R 1 is H, R 2 is the linker, and when R 2 is H, R 1 is the linker.

8. The method of claim 2 , wherein the hematopoietic stem cells or progeny thereof are capable of localizing to hematopoietic tissue and/or reestablishing hematopoiesis following transplantation of the hematopoietic stem cells into the patient.

9. The method of claim 7 , wherein the linker includes a —(CH 2 ) n — unit, wherein n is an integer from 2-6, or wherein the linker includes a dipeptide selected from Val-Ala and Val-Cit.

10. The method of claim 7 , wherein Am-L-Z-Ab is one of:

11. The method of claim 3 , wherein the antibody or antigen-binding fragment thereof,

comprises a V L having the amino acid sequence DIQMTQSPSSMSASLGDRVTITCRASQDINSYLSWFQQKPGKSPKTLIYRANRL VDGVPSRFSGSGSGTDYTLTISSLQYEDFGIYYCQQYDESPWTFGGGTKLEIK (SEQ ID NO: 1); and a V H having the amino acid sequence QIQLVQSGPGLKKPGGSVRISCAASGYTFTNYGMNWVKQAPGKGLRWMGWI NTHTGEPTYADDFKGRFTFSLDTSKSTAYLQINSLRAEDTATYFCTRRGYDWY FDVWGQGTTVTVSS (SEQ ID NO: 2); or

comprises a V L having the amino acid sequence NIVMTQSPSSLSASVGDRVTITCQASQDVGTAVAWYQQKPDQSPKLLIYWTSTRHTGVP DRFTGSGS GTDFTLTISSLQPEDIATYFCHQYNSYNTFGSGTKLEIK (SEQ ID NO: 258); and a V H having the amino acid sequence QVTLKESGPVLVKPTETLTLTCTFSGFSLSTSGMGVGWIRQAPGKGLEWVAHIWWDDD VYYNPSLKSRLTITKDASKDQVSLKLSSVTAADTAVYYCVRRRATGTGFDYWGQGTLV TVSS (SEQ ID NO: 257).

12. The method of claim 2 , wherein R A and R B , together with the oxygen atoms to which they are bound, combine to form a 5 membered heterocycloalkyl group of formula:

wherein Y is —C(—O)—, —C(═S)—, —C(═NR E )—, or —C(R E R E′ )—; and

R E and R E′ are each independently C 1 -C 6 alkylene-R C , C 1 -C 6 heteroalkylene-R C , C 2 -C 6 alkenylene-R C , C 2 -C 6 heteroalkenylene-R C , C 2 -C 6 alkynylene-R C , C 2 -C 6 heteroalkynylene-R C , cycloalkylene-R C , heterocycloalkylene-R C , arylene-R C , or heteroarylene-R C .

13. The method of claim 2 , wherein the antibody or antigen-binding fragment thereof conjugated to a cytotoxin is represented by the formula Ab-L-Z-Am, wherein Ab is the antibody or antigen-binding fragment thereof, Z is a chemical moiety, L is a linker, and Am is an amatoxin, and the amatoxin-linker conjugate Am-L-Z is represented by formula (IIB)

wherein X is S, SO, or SO 2 ;

R 1 is H or a linker covalently bound to the antibody or antigen-binding fragment thereof through a chemical moiety Z, formed from a coupling reaction between a reactive substituent present on the linker and a reactive substituent present within an antibody, or antigen-binding fragment thereof; and

R 2 is H or a linker covalently bound to the antibody or antigen-binding fragment thereof through a chemical moiety Z, formed from a coupling reaction between a reactive substituent present on the linker and a reactive substituent present within an antibody, or antigen-binding fragment thereof;

wherein when R 1 is H, R 2 is the linker, and when R 2 is H, R 1 is the linker.

Assignments (4)

Continuity (4)

References Cited (310)