FXa variant compositions
View Patent ↗Compositions are provided comprising recombinant variants of the human clotting Factor Xa. Such compositions include a wide variety of isoforms and post-translational modifications of FXa and are useful for treating subjects in need of hemostasis.
1. A composition with pro-coagulant activity comprising:
a first recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 386 of the amino acid sequence of SEQ ID NO:1;
and a second recombinant FXa variant protein selected from the group consisting of:
(i) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 139 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(ii) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(iii) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 142 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(iv) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 143 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(v) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 384 of the amino acid sequence of SEQ ID NO:1,
(vi) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 384 of the amino acid sequence of SEQ ID NO:1,
(vii) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 142 and 146 to 384 of the amino acid sequence of SEQ ID NO:1; and
(viii) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 143 and 146 to 384 of the amino acid sequence of SEQ ID NO:1.
2. The composition of claim 1 , wherein a light chain of at least one of said recombinant FXa variant proteins comprises β-hydroxy Asp 63 and an O-linked hexose.
3. The composition of claim 1 , wherein a light chain of at least one of said recombinant FXa variant proteins comprises 10 Gla residues.
4. The composition of claim 1 , wherein a light chain of at least one of said recombinant FXa variant proteins comprises 11 Gla residues.
5. The composition of claim 1 , wherein a light chain of at least one of said recombinant FXa variant proteins comprises β-hydroxy Asp 63 , an O-linked hexose, and 10 or 11 Gla residues.
6. The composition of claim 1 , wherein said composition comprises:
(i) a first recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(ii) a second recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 139 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(iii) a third recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 386 of the amino acid sequence of SEQ ID NO:1; and
(iv) a fourth recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 142 and 146 to 386 of the amino acid sequence of SEQ ID NO:1.
7. The composition of claim 6 , wherein a light chain of each of said first, second, third, and fourth recombinant FXa variant proteins comprises β-hydroxy Asp 63 , an O-linked hexose, and at least 10 Gla residues.
8. The composition of claim 7 , wherein a light chain of each of said first, third, and fourth recombinant FXa variant proteins comprises β-hydroxy Asp 63 , an O-linked hexose, and 11 Gla residues.
9. A composition with pro-coagulant activity comprising:
a first recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 140 and 146 to 398 of the amino acid sequence of SEQ ID NO:1;
and a second recombinant FXa variant protein selected from the group consisting of:
(i) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 139 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(ii) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 141 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(iii) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 141 and 146 to 399 of the amino acid sequence of SEQ ID NO:1,
(iv) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 142 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(v) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 399 of the amino acid sequence of SEQ ID NO:1, and
(vi) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 142 and 146 to 399 of the amino acid sequence of SEQ ID NO:1.
10. The composition of claim 9 , wherein a light chain of at least one of said recombinant FXa variant proteins comprises β-hydroxy Asp 63 and an O-linked hexose.
11. The composition of claim 9 , wherein a light chain of at least one of said recombinant FXa variant proteins comprises 10 Gla residues.
12. The composition of claim 9 , wherein a light chain of at least one of said recombinant FXa variant proteins comprises 11 Gla residues.
13. The composition of claim 9 , wherein a light chain of at least one of said recombinant FXa variant proteins comprises β-hydroxy Asp 63 , an O-linked hexose, and 10 or 11 Gla residues.
14. The composition of claim 9 , wherein a heavy chain of at least one of said recombinant FXa variant proteins comprises one core-1 O-linked glycan.
15. The composition of claim 14 , wherein the core-1 O-linked glycan is mono-sialylated.
16. The composition of claim 14 , wherein the core-1 O-linked glycan is di-sialylated.
17. The composition of claim 9 , wherein a heavy chain of at least one of said recombinant FXa variant proteins comprises two core-1 O-linked glycans.
18. The composition of claim 17 , wherein both core-1 O-linked glycans are mono-sialylated.
19. The composition of claim 17 , wherein both core-1 O-linked glycans are di-sialylated.
20. The composition of claim 17 , wherein one of said core-1 O-linked glycans is mono-sialylated and the second of said core-1 O-linked glycans is di-sialylated.
21. A composition with pro-coagulant activity comprising:
(i) a first recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 398 of the amino acid sequence of SEQ ID NO: 1,
(ii) a second recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 139 and 146 to 398 of the amino acid sequence of SEQ ID NO: 1,
(iii) a third recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 398 of the amino acid sequence of SEQ ID NO: 1; and
(iv) a fourth recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 399 of the amino acid sequence of SEQ ID NO: 1.
22. The composition of claim 21 , wherein a light chain of each of said first, second, third, and fourth recombinant FXa variant proteins comprises β-hydroxy Asp 63 , an O-linked hexose, and at least 10 Gla residues.
23. The composition of claim 22 , wherein a light chain of each of said first, second, third, and fourth recombinant FXa variant proteins comprises β-hydroxy Asp 63 , an O-linked hexose, and 11 Gla residues.
24. The composition of claim 21 , wherein a heavy chain of each of said first, second, third, and fourth recombinant FXa variant proteins comprises one core-1 O-linked glycan.
25. The composition of claim 24 , wherein the core-1 O-linked glycan is di-sialylated.
26. A composition with pro-coagulant activity comprising:
a first recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 399 of the amino acid sequence of SEQ ID NO:1;
and a second recombinant FXa variant protein selected from the group consisting of:
(i) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 o 399 of the amino acid sequence of SEQ ID NO:1; and
(ii) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 398 of the amino acid sequence of SEQ ID NO:1.
27. The composition of claim 26 , wherein a light chain of each of said first and second recombinant FXa variant proteins comprises β-hydroxy Asp 63 , an O-linked hexose, and at least 10 Gla residues.
28. The composition of claim 27 , wherein a light chain of each of said first and second recombinant FXa variant proteins comprises β-hydroxy Asp 63 , an O-linked hexose, and 11 Gla residues.
29. The composition of claim 26 , wherein a heavy chain of each of said first and second recombinant FXa variant proteins comprises two core-1 O-linked glycans.
30. The composition of claim 29 , wherein both core-1 O-linked glycans are di-sialylated.
31. A composition with pro-coagulant activity comprising two or more recombinant FXa variant proteins selected from the group consisting of:
(a) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 139 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(b) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 386 of the amino acid sequence of SEQ ID NO:1;
(c) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(d) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 142 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(e) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 143 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(f) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 384 of the amino acid sequence of SEQ ID NO:1,
(g) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 384 of the amino acid sequence of SEQ ID NO:1,
(h) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 142 and 146 to 384 of the amino acid sequence of SEQ ID NO:1, and
(i) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 143 and 146 to 384 of the amino acid sequence of SEQ ID NO:1.
32. The composition of claim 31 , wherein the light chains of said recombinant FXa variant proteins comprise β-hydroxy Asp 63 and an O-linked hexose.
33. The composition of claim 31 , wherein the light chains of said recombinant FXa variant proteins comprise β-hydroxy Asp 63 , an O-linked hexose, and 10 or 11 Gla residues.
34. The composition of claim 31 , comprising 3, 4, 5, 6, 7, 8, or all 9 recombinant FXa variant proteins selected from among (a)-(i).
35. A composition with pro-coagulant activity comprising two or more recombinant FXa variant proteins selected from the group consisting of:
(a) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 139 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(b) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 140 and 146 to 398 of the amino acid sequence of SEQ ID NO:1;
(c) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 141 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(d) A recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 142 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(e) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 399 of the amino acid sequence of SEQ ID NO:1,
(f) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 399 of the amino acid sequence of SEQ ID NO:1, and
(g) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 142 and 146 to 399 of the amino acid sequence of SEQ ID NO:1.
36. The composition of claim 35 , wherein the light chains of said recombinant FXa variant proteins comprise β-hydroxy Asp 63 and an O-linked hexose.
37. The composition of claim 35 , wherein the light chains of said recombinant FXa variant proteins comprise β-hydroxy Asp 63 , an O-linked hexose, and 10 or 11 Gla residues.
38. The composition of claim 35 , wherein the heavy chains of said recombinant FXa variant proteins comprise one or two core-1 O-linked glycans.
39. The composition of claim 38 , wherein at least one of said core-1 O-linked glycans is mono-sialylated or di-sialylated.
40. The composition of claim 35 , comprising 3, 4, 5, 6, or all 7 recombinant FXa variant proteins selected from among (a)-(g).
41. A composition with pro-coagulant activity comprising:
(a) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 139 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(b) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 386 of the amino acid sequence of SEQ ID NO:1;
(c) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(d) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 142 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(e) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 143 and 146 to 386 of the amino acid sequence of SEQ ID NO:1,
(f) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 384 of the amino acid sequence of SEQ ID NO:1,
(g) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 384 of the amino acid sequence of SEQ ID NO:1,
(h) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 142 and 146 to 384 of the amino acid sequence of SEQ ID NO:1,
(i) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 143 and 146 to 384 of the amino acid sequence of SEQ ID NO:1,
(j) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 139 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(k) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 140 and 146 to 398 of the amino acid sequence of SEQ ID NO:1;
(l) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 141 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(m) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 142 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(n) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 399 of the amino acid sequence of SEQ ID NO:1,
(o) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 399 of the amino acid sequence of SEQ ID NO:1, and
(p) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 142 and 146 to 399 of the amino acid sequence of SEQ ID NO:1.
42. The composition of claim 41 , wherein the light chains of said recombinant FXa variant proteins comprise β-hydroxy Asp 63 , an O-linked hexose, and 10 or 11 Gla residues.
43. The composition of claim 41 , wherein the heavy chains of said recombinant FXa variant proteins comprise one or two core-1 O-linked glycans.
44. The composition of claim 42 , wherein at least one of said core-1 O-linked glycans is mono-sialylated or di-sialylated.
45. The composition of claim 31 further comprising two or more recombinant FXa variant proteins selected from the group consisting of:
(a) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 139 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(b) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 140 and 146 to 398 of the amino acid sequence of SEQ ID NO:1;
(c) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 141 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(d) A recombinant FXa variant protein wherein the light chain and heavy chain protein sequence respectively consist of amino acids 1 to 142 and 146 to 398 of the amino acid sequence of SEQ ID NO:1,
(e) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 140 and 146 to 399 of the amino acid sequence of SEQ ID NO:1,
(f) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 141 and 146 to 399 of the amino acid sequence of SEQ ID NO:1, and
(g) a recombinant FXa variant protein wherein the light chain and heavy chain protein sequences respectively consist of amino acids 1 to 142 and 146 to 399 of the amino acid sequence of SEQ ID NO:1.