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KR-20260067387-A - Soluble bone morphogenetic protein (BMP) receptor type 1B protein and its uses

KR20260067387AKR 20260067387 AKR20260067387 AKR 20260067387AKR-20260067387-A

Abstract

The present invention generally relates to bone morphogenetic protein (BMP) antagonists (soluble ALK6 fusion proteins), compositions thereof, and methods used for treating neurodegenerative diseases and/or demyelinating diseases.

Inventors

  • 크놉프 존

Assignees

  • 래스크 파마, 인크.

Dates

Publication Date
20260512
Application Date
20240802
Priority Date
20230804

Claims (20)

  1. A soluble recombinant bone morphogenetic protein receptor type 1B fusion protein comprising an ALK6 extracellular domain (ECD) polypeptide and an Fc sequence, wherein the ALK6 ECD polypeptide comprises at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequence to the ALK6 ECD sequence selected from SEQ NO. 4; SEQ NO. 11, SEQ NO. 69; SEQ NO. 80 and SEQ NO. 113.
  2. A fusion protein according to claim 1, wherein the amino acid at position 60 is a glycine residue.
  3. In claim 1, the polypeptide is a fusion protein that binds to BMP10.
  4. A soluble recombinant bone morphogenetic protein receptor type 1B fusion protein comprising an ALK6 ECD polypeptide and an Fc sequence, wherein the ALK6 ECD polypeptide comprises an amino acid sequence selected from the sequence of SEQ ID NO. 3: Lys 1 - Lys 2 - Glu 3 - Asp 4 - Gly 5 - Glu 6 - Ser 7 - Thr 8 - Ala 9 - Pro 10 - Thr 11 - Pro 12 - Arg 13 -Pro 14 - Xaa 15 - Val 16 - Leu 17 - Arg 18 - Cys 19 - Xaa 20 - Cys 21 - Xaa 22 - Xaa 23 - His 24 - Cys 25 - Pro 26 - Xaa 27 - Asp 28 - Xaa 29 - Xaa 30 - Asn 31 - Asn 32 - Xaa 33 - Cys 34 - Xaa 35 - Thr 36 - 43 - Ile 44 - Glu 45 - Glu 46 - Asp 47 - Asp 48 - Xaa 49 - Gly 50 - Xaa 51 - Xaa 52 - Xaa 53 - Xaa 54 - 63 - Ser 64 - Asp 65 - Phe 66 - Gln 67 - Cys 68 - Xaa 69 - Asp 70 - Xaa 71 - Pro 72 - Xaa 73 - Xaa 74 - Xaa 75 - 82 - Cys 83 - Xaa 84 - Xaa 85 - Xaa 86 - Xaa 87 - Xaa 88 - Cys 89 - Asn 90 - Xaa 91 - Xaa 92 - Leu 93 - Xaa 102 - Xaa 103 - Xaa 104 - Xaa 105 - Xaa 106 - Xaa 107 - Xaa 108 - Xaa 109 - Here: Xaa 15 is K or P and; Xaa 20 is V, F, K or Y and; Xaa 22 is G, H or S and; Xaa 23 is S, H, or G; Xaa 27 is E or D and; Xaa 29 is S or A and; Xaa 30 is V or I; Xaa 33 is I or T and; Xaa 35 is S or I; Xaa 37 is D or N and; Xaa 39 is Y or H; Xaa 42 is T or A and; Xaa 43 is M or I; Xaa 49 is S or Q and; Xaa 51 is L or E and; Xaa 52 is P or T and; Xaa 53 is V or T and; Xaa 54 is V or L and; Xaa 55 is T or A and; Xaa 59 is L or M and; Xaa 60 is G or K and; Xaa 61 is L or Y and; Xaa 69 is R or K and; Xaa 71 is T or S and; Xaa 73 is I or K and; Xaa 74 is P or A and; Xaa 75 is H or Q; Xaa 76 is Q or L and; Xaa 79 is S or T and; Xaa 84 is T or R and; Xaa 85 is not, T or E and; Xaa 86 is R or N and; Xaa 87 is N or L and; Xaa 88 is not, E or L, and; Xaa 91 is K or Q and; Xaa 92 is D or L and; Xaa 94 is H or Q and; Xaa 100 is not, V or L; Xaa 101 is not, V or K, and; Xaa 102 is not, or is I or N; Xaa 103 is not, G or R, and; Xaa 104 is not, P, D, or A; Xaa 105 is not or is F; Xaa 106 is not, F or V, and; Xaa 107 is either non-existent or D; Xaa 108 is either non-existent or G; Xaa 109 is not, S or P, and; Xaa 110 is either non-existent or I; Xaa 111 is absent, R, or H; Xaa 112 is non-existent or H; and Xaa 113 is a fusion protein that is absent or R.
  5. A fusion protein according to claim 4, wherein the ALK6 ECD polypeptide comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 69%, 71%, 73%, 75%, 77%, 80%, 86%, 88%, 92%, 94%, 96%, 98%, and 100 identical to the ALK6 ECD sequence selected from sequence numbers 1, 4, 11-45, 57, 59, 61, 63, 65, 67%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%7%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ALK6 ECD sequence selected from sequence numbers 1, 4, 11-45, 57, 59
  6. In paragraph 4, the ALK6 ECD polypeptide comprises a substitution selected from K15P, K20Y, K20V, K20F, H22S, H22G, H23G, H23S, E27D, S29A, V30I, I33T, S35I, D37N, Y39H, T42A, M43I, S49Q, L51E, P52T, V53T, T55A, L59M, G60K, L61Y, R69K, T71S, I73K, P74A, H75Q, Q76L, S79T, T84R, E85T, E85_, R86N, N87L, E88_, K91Q, D92L, H94Q, D104A, D107, or a combination thereof, wherein _ is at the corresponding position Deletion, fusion protein.
  7. In paragraph 4, Xaa60 is a fusion protein in which glycine (G) residue is present.
  8. In paragraph 4, the polypeptide is a fusion protein that binds to BMP10.
  9. A soluble recombinant bone morphogenetic protein receptor type 1B fusion protein comprising an ALK6 ECD polypeptide and an Fc sequence, wherein the ALK6 ECD polypeptide comprises an amino acid sequence selected from the sequence of SEQ ID NO. 5: Lys 1 - Lys 2 - Glu 3 - Asp 4 - Gly 5 - Glu 6 - Ser 7 - Thr 8 - Ala 9 - Pro 10 - Thr 11 - Pro 12 - Arg 13 -Pro 14 - Xaa 15 - Val 16 - Leu 17 - Arg 18 - Cys 19 - Xaa 20 - Cys 21 - Xaa 22 - Xaa 23 - His 24 - Cys 25 - Pro 26 - Xaa 27 - Asp 28 - Xaa 29 - Xaa 30 - Asn 31 - Asn 32 - Xaa 33 - Cys 34 - Xaa 35 - Thr 36 - 43 - Ile 44 - Glu 45 - Glu 46 - Asp 47 - Asp 48 - Xaa 49 - Gly 50 - Xaa 51 - Xaa 52 - Xaa 53 - Xaa 54 - 63 - Ser 64 - Asp 65 - Phe 66 - Gln 67 - Cys 68 - Xaa 69 - Asp 70 - Xaa 71 - Pro 72 - Xaa 73 - Xaa 74 - Xaa 75 - 82 - Cys 83 - Xaa 84 - Xaa 85 - Xaa 86 - Xaa 87 - Xaa 88 - Cys 89 - Asn 90 - Xaa 91 - Xaa 92 - Leu 93 - Here, Xaa 15 is K or P and; Xaa 20 is V, F, K or Y and; Xaa 22 is G, H or S and; Xaa 23 is S, H, or G; Xaa 27 is E or D and; Xaa 29 is S or A and; Xaa 30 is V or I; Xaa 33 is I or T and; Xaa 35 is S or I; Xaa 37 is D or N and; Xaa 39 is Y or H; Xaa 42 is T or A and; Xaa 43 is M or I; Xaa 49 is S or Q and; Xaa 51 is L or E and; Xaa 52 is P or T and; Xaa 53 is V or T and; Xaa 54 is V or L and; Xaa 55 is T or A and; Xaa 59 is L or M and; Xaa 60 is G or K and; Xaa 61 is L or Y and; Xaa 69 is R or K and; Xaa 71 is T or S and; Xaa 73 is I or K and; Xaa 74 is P or A and; Xaa 75 is H or Q; Xaa 76 is Q or L and; Xaa 79 is S or T and; Xaa 84 is T or R and; Xaa 85 is not, T or E and; Xaa 86 is R or N and; Xaa 87 is N or L and; Xaa 88 is not, E or L, and; Xaa 91 is K or Q and; Xaa 92 is D or L; and, Xaa 94 is a fusion protein that is H or Q.
  10. In claim 9, the ALK6 ECD polypeptide comprises a substitution selected from K15P, K20Y, K20V, K20F, H22S, H22G, H23G, H23S, E27D, S29A, V30I, I33T, S35I, D37N, Y39H, T42A, M43I, S49Q, L51E, P52T, V53T, T55A, L59M, G60K, L61Y, R69K, T71S, I73K, P74A, H75Q, Q76L, S79T, T84R, E85T, E85_, R86N, N87L, E88_, K91Q, D92L, H94Q, or a combination thereof, wherein _ is a deletion at the corresponding position, a fusion protein.
  11. In claim 9, Xaa60 is a fusion protein that is a glycine (G) residue.
  12. In claim 9, the polypeptide is a fusion protein that binds to BMP10.
  13. A soluble recombinant bone morphogenetic protein receptor type 1B fusion protein comprising an ALK6 ECD polypeptide and an Fc sequence, wherein the ALK6 ECD polypeptide comprises an amino acid sequence selected from the sequence of SEQ ID NO. 79: Lys 1 - Lys 2 - Glu 3 - Asp 4 - Gly 5 - Glu 6 - Ser 7 - Thr 8 - Ala 9 - Pro 10 - Thr 11 - Pro 12 - Arg 13 -Pro 14 - Xaa 15 - Val 16 - Leu 17 - Arg 18 - Cys 19 - Xaa 20 - Cys 21 - Xaa 22 - Xaa 23 - His 24 - Cys 25 - Pro 26 - Xaa 27 - Asp 28 - Xaa 29 - Xaa 30 - Asn 31 - Asn 32 - Xaa 33 - Cys 34 - Xaa 35 - Thr 36 - 43 - Ile 44 - Glu 45 - Glu 46 - Asp 47 - Asp 48 - Xaa 49 - Gly 50 - Xaa 51 - Xaa 52 - Xaa 53 - Xaa 54 - 63 - Ser 64 - Asp 65 - Phe 66 - Gln 67 - Cys 68 - Xaa 69 - Asp 70 - Xaa 71 - Pro 72 - Xaa 73 - Xaa 74 - Xaa 75 - 82 - Cys 83 - Xaa 84 - Xaa 85 - Xaa 86 - Xaa 87 - Xaa 88 - Cys 89 - Asn 90 - Xaa 91 - Xaa 92 - Leu 93 - -Ile 102 -Gly 103 -Pro 104 -Phe 105 -Phe 106 -Asp 107 -Gly 108 -Ser 109 -Ile 110 -Arg 111 ; Here, Xaa 15 is K or P and; Xaa 20 is V, F, K or Y and; Xaa 22 is G, H or S and; Xaa 23 is S, H, or G; Xaa 27 is E or D and; Xaa 29 is S or A and; Xaa 30 is V or I; Xaa 33 is I or T and; Xaa 35 is S or I; Xaa 37 is D or N and; Xaa 39 is Y or H; Xaa 42 is T or A and; Xaa 43 is M or I; Xaa 49 is S or Q and; Xaa 51 is L or E and; Xaa 52 is P or T and; Xaa 53 is V or T and; Xaa 54 is V or L and; Xaa 55 is T or A and; Xaa 59 is L or M and; Xaa 60 is G or K and; Xaa 61 is L or Y and; Xaa 69 is R or K and; Xaa 71 is T or S and; Xaa 73 is I or K and; Xaa 74 is P or A and; Xaa 75 is H or Q; Xaa 76 is Q or L and; Xaa 79 is S or T and; Xaa 84 is T or R and; Xaa 85 is not, T or E, and; Xaa 86 is R or N and; Xaa 87 is N or L and; Xaa 88 is not, E or L, and; Xaa 91 is K or Q and; Xaa 92 is D or L; and, Xaa 94 is a fusion protein that is H or Q.
  14. A fusion protein according to claim 13, wherein the ALK6 ECD polypeptide comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ALK6 ECD sequence selected from sequence numbers 15-45, 57, 59, 61, 63, 65, and 88.
  15. In claim 13, the ALK6 ECD polypeptide comprises a substitution selected from K15P, K20Y, K20V, K20F, H22S, H22G, H23G, H23S, E27D, S29A, V30I, I33T, S35I, D37N, Y39H, T42A, M43I, S49Q, L51E, P52T, V53T, T55A, L59M, G60K, L61Y, R69K, T71S, I73K, P74A, H75Q, Q76L, S79T, T84R, E85T, E85_, R86N, N87L, E88_, K91Q, D92L, H94Q, D104A, D107, or a combination thereof, wherein _ represents the corresponding A fusion protein that is a deletion at a specific location.
  16. A soluble recombinant bone morphogenetic protein receptor type 1B fusion protein comprising an ALK6 ECD polypeptide and an Fc sequence, wherein the ALK6 ECD polypeptide comprises an amino acid sequence selected from the sequence of SEQ ID NO. 81: Lys 1 - Lys 2 - Glu 3 - Asp 4 - Gly 5 - Glu 6 - Ser 7 - Thr 8 - Ala 9 - Pro 10 - Thr 11 - Pro 12 - Arg 13 -Pro 14 - Xaa 15 - Val 16 - Leu 17 - Arg 18 - Cys 19 - Xaa 20 - Cys 21 - Xaa 22 - Xaa 23 - His 24 - Cys 25 - Pro 26 - Xaa 27 - Asp 28 - Xaa 29 - Xaa 30 - Asn 31 - Asn 32 - Xaa 33 - Cys 34 - Xaa 35 - Thr 36 - 43 - Ile 44 - Glu 45 - Glu 46 - Asp 47 - Asp 48 - Xaa 49 - Gly 50 - Xaa 51 - Xaa 52 - Xaa 53 - Xaa 54 - 63 - Ser 64 - Asp 65 - Phe 66 - Gln 67 - Cys 68 - Xaa 69 - Asp 70 - Xaa 71 - Pro 72 - Xaa 73 - Xaa 74 - Xaa 75 - 82 - Cys 83 - Xaa 84 - Xaa 85 - Xaa 86 - Xaa 87 - Xaa 88 - Cys 89 - Asn 90 - Xaa 91 - Xaa 92 - Leu 93 - Asn 102 - Arg 103 - Asp 104 - Phe 105 - Val 106 - Xaa 107 - Xaa 108 - Xaa 109 - Xaa 110 - Xaa 111 - Xaa 112 - Xaa 113 ; Here, Xaa 15 is K or P and; Xaa 20 is V, F, K or Y and; Xaa 22 is G, H or S and; Xaa 23 is S, H, or G; Xaa 27 is E or D and; Xaa 29 is S or A and; Xaa 30 is V or I; Xaa 33 is I or T and; Xaa 35 is S or I; Xaa 37 is D or N and; Xaa 39 is Y or H; Xaa 42 is T or A and; Xaa 43 is M or I; Xaa 49 is S or Q and; Xaa 51 is L or E and; Xaa 52 is P or T and; Xaa 53 is V or T and; Xaa 54 is V or L and; Xaa 55 is T or A and; Xaa 59 is L or M and; Xaa 60 is G or K and; Xaa 61 is L or Y and; Xaa 69 is R or K and; Xaa 71 is T or S and; Xaa 73 is I or K and; Xaa 74 is P or A and; Xaa 75 is H or Q; Xaa 76 is Q or L and; Xaa 79 is S or T and; Xaa 84 is T or R and; Xaa 85 is not, T or E, and; Xaa 86 is R or N and; Xaa 87 is N or L and; Xaa 88 is not, E or L, and; Xaa 91 is K or Q and; Xaa 92 is D or L and; Xaa 94 is H or Q and; Xaa 107 is not there; There is no Xaa 108 ; There is no Xaa 109 ; There is no Xaa 110 ; Xaa 111 is not there; Xaa 112 is not; and, Xaa 113 is a non-existent fusion protein.
  17. A fusion protein according to claim 16, wherein the ALK6 ECD polypeptide comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ALK6 ECD sequence selected from sequence numbers 13, 86, 92, 94%, 96%, 97%, 98%, 99%, or 100% identical to the ALK6 ECD sequence selected from sequence numbers 13, 86, 92, 94, 96%, 98%, 98%, or 100% identical.
  18. In claim 16, the ALK6 ECD polypeptide comprises a substitution selected from K15P, K20Y, K20V, K20F, H22S, H22G, H23G, H23S, E27D, S29A, V30I, I33T, S35I, D37N, Y39H, T42A, M43I, S49Q, L51E, P52T, V53T, T55A, L59M, G60K, L61Y, R69K, T71S, I73K, P74A, H75Q, Q76L, S79T, T84R, E85T, E85_, R86N, N87L, E88_, K91Q, D92L, H94Q, D104A, or a combination thereof, wherein _ is a deletion at the corresponding position, fusion protein.
  19. In paragraph 16, Xaa60 is a fusion protein that is a glycine (G) residue.
  20. In paragraph 16, the polypeptide is a fusion protein that binds to BMP10.

Description

Soluble bone morphogenetic protein (BMP) receptor type 1B protein and its uses Related applications This application claims priority to U.S. provisional application No. 63/530,928 filed on August 4, 2023, the entire contents of which are incorporated by reference into this application. Sequence list This application includes a list of sequences electronically submitted in XML format through the United States Patent and Trademark Office Patent Center, the full text of which is incorporated herein by reference. The name of the XML copy created on August 1, 2024 is 1902004-0002-001-101_SL.xml and the size is 240,412 bytes. Field of invention The present application generally relates to bone morphogenetic protein (BMP) antagonists, compositions thereof, and methods used for the treatment of BMP-related diseases. Background of the Invention The immunopathological mechanism of multiple sclerosis (MS) is characterized by pathogenic T cells and B cells infiltrating the central nervous system to induce multiple inflammation, demyelination, and neurodegeneration. Mature oligodendrocytes form myelin sheaths on axons in the CNS. The development of myelin depends on the proper maturation of oligodendrocytes from progenitor cells, and this is a spatially restricted process regulated by inductive and inhibitory signals. Several members of the bone morphogenetic protein (BMP) family play various roles in oligodendrocyte development, particularly by influencing cell lineage determination in oligodendrocyte progenitors (e.g., acting as inhibitors to convert oligodendrocyte progenitors into the astrocyte lineage) and regulating myelin protein expression in immature oligodendrocytes; in later stages, BMPs inhibit cell differentiation. For instance, BMPs have been shown to disrupt myelination by blocking the expression of various myelin structural proteins. BMPs also inhibit oligodendrocyte development and promote astrogenesis at various stages within their respective lineages. Myelination in the CNS by oligodendrocytes derived from progenitor cells that have undergone multiple phenotypic stages ensures the rapid and efficient transmission of electrical signals and enhances neuronal health and survival. The transforming growth factor beta (TGF-beta) superfamily includes various growth factors that share common sequence elements and structural motifs. These proteins are known to exert biological effects on a wide variety of cell types in both vertebrates and invertebrates. Members of this superfamily perform important functions in pattern formation and tissue characterization during embryonic development and can influence various differentiation processes, including adipogenesis, myogenesis, osteoogenesis, chondrogenesis, cardiogenesis, hematopoiesis, neurogenesis, and epithelial differentiation. This family is broadly divided into two phylogenetic clades: one is a clade of relatively recently evolved superfamily members including TGF-beta, activin, and nodals, and the other is a clade of relatively distantly related proteins within the superfamily that includes numerous BMPs and GDFs. Hinck (2012) FEBS Letters 586:1860-1870. Members of the TGF-beta family exhibit diverse and often complementary biological effects. Manipulating the activity of members of the TGF-beta family can often induce significant physiological changes in organisms. For example, Piedmontese and Belgian Blouse cattle possess loss-of-function mutations in the GDF8 (also known as myostatin) gene, resulting in a significant increase in muscle mass. Grobet et al. (1997) Nat Genet., 17(1):71-4. Furthermore, in humans, the inactive allele of GDF8 is known to be associated with increased muscle mass and superior muscle strength. Schuelke et al. (2004) N Engl J Med, 350:2682-8. Changes in muscle, bone, fat, red blood cells, and other tissues can be achieved by enhancing or inhibiting signal transduction mediated by TGF-beta family ligands (e.g., SMAD 1, 2, 3, 5, and/or 8). Therefore, there is a need for a formulation that regulates the activity of various ligands of the TGF-beta superfamily, including bone morphogenetic proteins (BMPs) related to the present disclosure. Summary of the Invention The present invention describes polypeptides, compositions, other reagents and/or BMP ligands related to bone morphogenetic protein (BMP) antagonists and methods for use in the treatment of neurodegenerative and/or demyelinating diseases associated with endogenous signaling. In particular, the present disclosure relates to soluble bone morphogenetic protein (BMP) receptor type 1B polypeptides (“ALK6 polypeptides”) comprising the extracellular domain of ALK6 (activin receptor-like kinase 6) and soluble variants thereof, which bind to BMP ligands to deprive the ligands of binding to the receptors (“BMP antagonists”). The ALK-6 polypeptides of the present disclosure bind to BMP ligands comprising, but not limited to, BMP 4 and/or 10. In certain embodiments, the ALK-6 polypeptide