BISPECIFIC ANTIBODIES AGAINST CD3EPSILON AND BCMA

• US 20170306036A1
• Filed: 08/03/2015
• Published: 10/26/2017
• Est. Priority Date: 08/04/2014
• Status: Active Grant

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: EngMab AG (Bristol-Myers Squibb Company)
• Inventors: VU, Minh Diem, STREIN, Klaus, AST, Oliver, BACAC, Marina, HANISCH, Lydia Jasmin, FAUTI, Tanja, FREIMOSER-GRUNDSCHOBER, Anne, HOSSE, Ralf, KLEIN, Christian, MOESSNER, Ekkehard, MOSER, Samuel, MURR, Ramona, UMANA, Pablo, JUNG-IMHOF, Sabine, KLOSTERMANN, Stefan, MOLHOJ, Michael, REGULA, Joerg, SCHAEFER, Wolfgang
• Granted Patent: US 10,253,104 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 16/3061 Blood cells
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/35 Valency
• CPC Class Codes: C07K 2317/522 CH1 domain
• CPC Class Codes: C07K 2317/526 CH3 domain
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/64 comprising a combination of variable region and constant region components
• CPC Class Codes: C07K 2317/66 comprising a swap of domains, e.g. CH3-CH2, VH-CL or VL-CH1
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

A bispecific bi- or trivalent antibody specifically binding to the two targets which are extracellular domain of human B cell maturation antigen (BCMA) and human CD3epsilon, wherein the variable domains VL and VH in a light chain and the respective heavy chain are replaced by each other, characterized in comprising a constant domain CL wherein the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and in the respective constant domain CH1 the amino acid at position 147 and the amino acid at position 213 is substituted independently by glutamic acid (E), r aspartic acid (D) (numbering according to Kabat). Also the manufacture and use of said antibody.

Claims

1. A bispecific bi- or trivalent antibody specifically binding to the two targets which are the extracellular domain of human B cell maturation antigen (further named also as “

BCMA”

) and human CD3ε

(further named also as “

CD3”

) wherein the variable domains VL and VH in a light chain and the respective heavy chain are replaced by each other, characterized in comprising a constant domain CL wherein the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and in the respective constant domain CH1 the amino acid at position 147 and the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat).

11. A method for the preparation of an a bispecific antibody according to claim 1 comprising the steps of

a) transforming a host cell with vectors comprising nucleic acid molecules encoding the light chain and heavy chain,

b) culturing the host cell under conditions that allow synthesis of said antibody molecule;

and

c) recovering said antibody molecule from said culture.

12. A host cell comprising vectors comprising nucleic acid molecules encoding the light chain and heavy chains of an antibody according to claim 1.

13. A pharmaceutical composition comprising an antibody according to claim 1 and a pharmaceutically acceptable excipient.

16. A method of treatment of a plasma cell disorder in a subject comprising administering an effective amount of composition according to claim 13 to said subject.

17. A method of treatment of Multiple Myeloma according to claim 16.

2. A bispecific antibody specifically binding to the two targets which are the extracellular domain of human BCMA and human CD3, characterized in comprising

a) the first light chain and the first heavy chain of a first antibody which specifically binds to BCMA;

and

b) the second light chain and the second heavy chain of a second antibody which specifically binds to CD3, and wherein the variable domains VL and VH in the second light chain and second heavy chain of the second antibody are replaced by each other; and

wherein

c) in the constant domain CL of the first light chain under a) the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and wherein in the constant domain CH1 of the first heavy chain under a) the amino acid at position 147 and the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat).

4. A bispecific antibody according to claim 2, characterized in that said bispecific antibody comprises in addition a Fab fragment of said first antibody (further named also as “

BCMA-Fab”

) and in the constant domain CL said BCMA-Fab the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and wherein in the constant domain CH1 of said BCMA-Fab the amino acid at positions 147 and the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat).

3. A bispecific antibody specifically binding to the two targets which are the extracellular domain of human BCMA and human CD3, characterized in comprising

a) the first light chain and the first heavy chain of a first antibody which specifically binds to BCMA;

and

b) the second light chain and the second heavy chain of a second antibody which specifically binds to CD3, and wherein the variable domains VL and VH in the second light chain and second heavy chain of the second antibody are replaced by each other; and

wherein

c) in the constant domain CL of the second light chain under b) the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and wherein in the constant domain CH1 of the second heavy chain under b) the amino acid at positions 147 and the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat).

5. A bispecific antibody according to claim 3, characterized in that said bispecific antibody comprises in addition a second Fab fragment of said first antibody (“

BCMA-Fab”

).

6. A bispecific antibody according to claim 5, characterized in that in addition to the amino acid replacement at position 124 in the constant domain CL the amino acid at position 123 is substituted independently by lysine (K), arginine (R) or histidine (H).

7. A bispecific antibody according to claim 6, characterized in that amino acid 124 is K, amino acid 147 is E, amino acid 213 is E, and amino acid 123 is R.

10. An antibody according to claim 7, characterized in that the CH3 domain of one heavy chain and the CH3 domain of the other heavy chain each meet at an interface which comprises an original interface between the antibody CH3 domains;

wherein said interface is altered to promote the formation of the bispecific antibody, wherein the alteration is characterized in that;

a) the CH3 domain of one heavy chain is altered, so that within the original interface the CH3 domain of one heavy chain that meets the original interface of the CH3 domain of the other heavy chain within the bispecific antibody, an amino acid residue is replaced with an amino acid residue having a larger side chain volume, thereby generating a protuberance within the interface of the CH3 domain of one heavy chain which is positionable in a cavity within the interface of the CH3 domain of the other heavy chain and

b) the CH3 domain of the other heavy chain is altered, so that within the original interface of the second CH3 domain that meets the original interface of the first CH3 domain within the bispecific antibody an amino acid residue is replaced with an amino acid residue having a smaller side chain volume, thereby generating a cavity within the interface of the second CH3 domain within which a protuberance within the interface of the first CH3 domain is positionable.

8. A bispecific antibody specifically binding to the extracellular domain of human BCMA and to human CD3, characterized in comprising a heavy and light chain set selected from the group consisting of polypeptides

i) SEQ ID NO:

43, SEQ ID NO;

44, SEQ ID NO;

45, and SEQ ID NO;

46 (set

     1),

ii) SEQ ID NO;

45, SEQ ID NO;

47, SEQ ID NO;

48, and SEQ ID NO;

49 (set

     2), and

iii) SEQ ID NO;

45, SEQ ID NO;

50, SEQ ID NO;

51, and SEQ ID NO;

52 (set

     3).

9. An antibody according to claim 8, characterized in that in the antibody portion specifically binding to human CD3 the variable domain VH is replaced by a variable domain VH comprising the heavy chain CDRs of SEQ ID NO:

1, 2 and 3 as respectively heavy chain CDR1, CDR2 and CDR3 and the variable domain VL is replaced by a variable domain VL comprising the light chain CDRs of SEQ ID NO;

4, 5 and 6 as respectively light chain CDR1, CDR2 and CDR3 of the anti CD3ε

antibody.

14-15. -15. (canceled)