IP Library › Granted Patent US 12686720

Granted Patent B2

US 12686720 · App. 19/297,899 · Granted Jul 21, 2026

Bispecific agonistic antibodies to activin a receptor like type 1 (ALK1)

Inventors: Alexey Alexandrovich Lugovskoy (Belmont, MA); Jean-Christophe Hus (Concord, MA); Melissa Geddie (Arlington, MA)

Assignee: DIAGONAL THERAPEUTICS INC.

C07K16/2863 A61P3/00 C07K16/2896 A61K2039/505 C07K2317/31 C07K2317/53 C07K2317/565 C07K2317/622 C07K2317/75 C07K2317/94

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Quick Facts

Patent No.: US 12686720
App. No.: 19/297,899
Granted: Jul 21, 2026
Kind: B2

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Abstract

Provided herein are bispecific agonistic antibodies that bind to ALK1, BMPRII, ActRIIA, and/or ActRIIB, and methods of using the same.

Claims (63)

1 . A multispecific binding protein comprising at least a first polypeptide chain and a second polypeptide chain, wherein said first polypeptide chain comprises a first variable heavy chain domain (VH1) linked to a second variable heavy chain domain (VH2) via at least one modified hinge region and wherein said second polypeptide chain comprises a first variable light chain domain (VL1) linked to a second variable light chain domain (VL2), wherein:

the VH1 binds specifically to BMPRII and the VH2 binds specifically to ALK1; or

the VH1 binds specifically to ALK1 and the VH2 binds specifically to BMPRII;

and wherein

the VH1 or VH2 binding to ALK1 comprises an HCDR1 amino acid sequence of SYWMS (SEQ ID NO:164), an HCDR2 amino acid sequence of NINQDGSEKYYVDSMRG (SEQ ID NO:165), and an HCDR3 amino acid sequence of EYDY (SEQ ID NO:166); and the VL binding to ALK1 comprises an LCDR1 amino acid sequence of SGSSSNIGSNYVY (SEQ ID NO:161), an LCDR2 amino acid sequence of GNNKRPS (SEQ ID NO:162), and an LCDR3 amino acid sequence of AAWDDSLNGRV (SEQ ID NO: 163); and

the VH1 or VH2 binding to BMPRII comprises an HCDR1 amino acid sequence of DYYMN (SEQ ID NO:175), an HCDR2 amino acid sequence of SISGGSTYYADSVKG (SEQ ID NO: 176), and an HCDR3 amino acid sequence of DFGVAGWFGQFGMDV (SEQ ID NO: 177); and the VL binding to BMPRII comprises an LCDR1 amino acid sequence of TGSSSNIGAGYDVH (SEQ ID NO:172), an LCDR2 amino acid sequence of RSNQRPS (SEQ ID NO: 173), and an LCDR3 amino acid sequence of SSYAGNYNLV (SEQ ID NO:174).

2 . The multispecific binding protein of claim 1 , wherein the VL1 is linked to the VL2 via at least one modified hinge region.

3 . The multispecific binding protein of claim 1 , wherein one or both of VH1 and VH2 is truncated at the C-terminal end.

4 . The multispecific binding protein of claim 3 , wherein the C-terminal end is truncated by at least one residue.

5 . The multispecific binding protein of claim 3 , wherein the C-terminal end is truncated by at least two residues.

6 . The multispecific binding protein of claim 3 , wherein the SS amino acid residues of the C-terminal end are deleted.

7 . The multispecific binding protein of claim 1 , wherein the first polypeptide chain is of the formula VH1-HX1-VH2-C-Fc, wherein:

VH1 is the first heavy chain variable domain;

VH2 is the second heavy chain variable domain;

C is a heavy chain constant domain;

HX1 is a modified hinge region linker; and

Fc is an Fc region; and

the second polypeptide chain is of the formula VL1-LX1-VL2-C,

wherein:

VL1 is the first light chain variable domain;

VL2 is v second light chain variable domain;

C is a light chain constant domain; and

LX1 is a modified hinge region linker.

8 . The multispecific binding protein of claim 1 , wherein the modified hinge region comprises; i) an upper hinge region of up to 7 amino acids in length or is absent; and ii) a lower hinge region.

9 . The multispecific binding protein of claim 1 , wherein the modified hinge region comprises or consists of an amino acid sequence of PLAP (SEQ ID NO:2) or PAPNLLGGP (SEQ ID NO:157).

10 . The multispecific binding protein of claim 1 , comprising at least a first polypeptide chain and a second polypeptide chain, wherein said first polypeptide chain comprises a first variable heavy chain domain (VH1) linked to a second variable heavy chain domain (VH2) and wherein said second polypeptide chain comprises a first variable light chain domain (VL1) linked to a second variable light chain domain (VL2), wherein:

the VH1 binds specifically to BMPRII and the VH2 binds specifically to ALK1; or

the VH1 binds specifically to ALK1 and the VH2 binds specifically to BMPRII and, wherein:

the VH binding to ALK1 comprises an amino acid sequence of EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVANINQDGSEK YYVDSMRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREYDYWGQGTLVTVSS (SE Q ID NO: 182), or an amino acid sequence with at least 90% identity thereto; and the VL binding to ALK1 comprises an amino acid sequence of QSVLAQPPSASGTPGQRVTISCSGSSSNIGSNYVYWYQQLPGTAPKLLIYGNNKRPSGVP DRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGRVFGGGTKLTVL (SEQ ID NO: 181), or an amino acid sequence with at least 90% identity thereto; and/or

the VH binding to BMPRII comprises an amino acid sequence of EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMNWIRQAPGKGLEWVSSISGGSTYYA DSVKGRFTISRDNSENTLYLQMNSLRAEDTAVYYCARDFGVAGWFGQFGMDVWGQGT LVTVSS (SEQ ID NO:186), or an amino acid sequence with at least 90% identity thereto; and

the VL binding to BMPRII comprises an amino acid sequence of QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYRSNQRPSGV PDRFSGSKSGTSASLAISGLRSEDEADYYCSSYAGNYNLVFGGGTKLTVL (SEQ ID NO: 185), or an amino acid sequence with at least 90% identity thereto.

11 . The multispecific binding protein of claim 10 , wherein:

12 . The multispecific binding protein of claim 1 , wherein the first polypeptide chain comprises an amino acid sequence of SEQ ID NO: 139, and the second polypeptide chain comprises an amino acid sequence of SEQ ID NO: 146.

13 . A multispecific binding protein comprising a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain and second polypeptide chain each comprise, from N-terminus to C-terminus, a first single chain variable fragment (scFv) linked to a second scFv, wherein:

the first scFv binds specifically to BMPRII and the second scFv binds specifically to ALK1; or

the first scFv binds specifically to ALK1 and the second scFv binds specifically to BMPRII;

wherein the scFv binding to ALK1 comprises a VH domain and a VL domain the VH domain comprising an HCDR1 amino acid sequence of SYWMS (SEQ ID NO:164), an HCDR2 amino acid sequence of NINQDGSEKYYVDSMRG (SEQ ID NO:165), and an HCDR3 amino acid sequence of EYDY (SEQ ID NO:166); and the VL domain comprising an an LCDR1 amino acid sequence of SGSSSNIGSNYVY (SEQ ID NO:161), an LCDR2 amino acid sequence of GNNKRPS (SEQ ID NO:162), and an LCDR3 amino acid sequence of AAWDDSLNGRV (SEQ ID NO:163) and

wherein the scFv binding to BMPRII comprises a VH domain comprising an HCDR1 amino acid sequence of DYYMN (SEQ ID NO:175), an HCDR2 amino acid sequence of SISGGSTYYADSVKG (SEQ ID NO:176), and an HCDR3 amino acid sequence of DFGVAGWFGQFGMDV (SEQ ID NO:177); and a VL domain comprising an LCDR1 amino acid sequence of TGSSSNIGAGYDVH (SEQ ID NO:172), an LCDR2 amino acid sequence of RSNQRPS (SEQ ID NO:173), and an LCDR3 amino acid sequence of SSYAGNYNLV (SEQ ID NO: 174).

14 . The multispecific binding protein of claim 13 , wherein the first scFv is linked to the second scFv via at least one modified hinge region.

15 . The multispecific binding protein of claim 1 , wherein the first polypeptide chain further comprises a heavy chain constant region.

16 . The multispecific binding protein of claim 15 , wherein the heavy chain constant region comprises an alanine (A) at amino acid positions 234, 235, and/or 237, according to EU numbering.

17 . A multispecific binding protein comprising at least a first binding moiety and a second binding moiety, wherein said first binding moiety comprises a first variable heavy chain domain (VH1) and first variable light chain domain (VL1), and the second binding moiety comprising a second variable heavy chain domain (VH2) and a second variable light chain domain (VL2), wherein:

the VH1 binds specifically to BMPRII and the VH2 binds specifically to ALK1; or

the VH1 binds specifically to ALK1 and the VH2 binds specifically to BMPRII;

and wherein

the VH1 or VH2 binding to BMPRII comprises an HCDR1 amino acid sequence of DYYMN (SEQ ID NO:175), an HCDR2 amino acid sequence of SISGGSTYYADSVKG (SEQ ID NO: 176), and an HCDR3 amino acid sequence of DFGVAGWFGQFGMDV (SEQ ID NO: 177); and the VL1 or VL2 binding to BMPRII comprises an LCDR1 amino acid sequence of TGSSSNIGAGYDVH (SEQ ID NO:172), an LCDR2 amino acid sequence of RSNQRPS (SEQ ID NO: 173), and an LCDR3 amino acid sequence of SSYAGNYNLV (SEQ ID NO:174), and

wherein the multispecific binding protein further comprises a heavy chain constant region wherein the heavy chain constant region comprises heterodimerization mutations to promote heterodimerization of the first binding moiety with the second binding moiety.

18 . The multispecific binding protein of claim 17 , wherein the heterodimerization mutations are Knob-in-Hole (KIH) mutations.

19 . The multispecific binding protein of claim 18 , wherein the first heavy chain constant region comprises an amino acid substitution at position 366, 368, or 407 which produced a hole, and the second heavy chain constant region comprises an amino acid substitution at position 366 which produce a knob.

20 . The multispecific binding protein of claim 19 , wherein the first heavy chain constant region comprises the amino acid substitution T366S, L368A, or Y407V, and the second heavy chain constant region comprises the amino acid substitution T366W.

21 . The multispecific binding protein of claim 17 , wherein the heterodimerization mutations are charge stabilization mutations.

22 . The multispecific binding protein of claim 21 , wherein the first heavy chain constant region comprises the amino acid substitution N297K, and the second heavy chain constant region comprises the amino acid substitution N297D.

23 . The multispecific binding protein of claim 21 , wherein the first heavy chain constant region comprises the amino acid substitution T299K, and the second heavy chain constant region comprises the amino acid substitution T299D.

24 . The multispecific binding protein of claim 17 , wherein the heterodimerization mutations comprise an engineered disulfide bond.

25 . The multispecific binding protein of claim 24 , wherein the engineered disulfide bond is formed by a first heavy chain constant region comprising the amino acid substitution Y349C, and a second heavy chain constant region comprising the amino acid substitution S354C.

26 . The multispecific binding protein of claim 24 , wherein the engineered disulfide bond is formed by a C-terminal extension peptide fused to the C-terminus of each of the first heavy chain constant region and the second heavy chain constant region.

27 . The multispecific binding protein of claim 17 , wherein at least one heavy chain constant region comprises one or more mutations to promote increased half-life.

28 . The multispecific binding protein of claim 27 , wherein at least one heavy chain constant region comprises one or more substitutions at amino acid positions 428 or 434, according to EU numbering.

29 . The multispecific binding protein of claim 28 , wherein at least one heavy chain constant region comprises a M428L and N434S substitution, according to EU numbering.

30 . A pharmaceutical composition comprising the multispecific binding protein of claim 1 and a pharmaceutically acceptable carrier.