COAGULATION FACTOR VII COMPOSITIONS AND METHODS OF MAKING AND USING SAME
The present invention relates to compositions comprising factor VII coagulation factors linked to extended recombinant polypeptide (XTEN), isolated nucleic acids encoding the compositions and vectors and host cells containing the same, and methods of making and using such compositions in treatment of coagulation factor-related diseases, disorders, and conditions.
1 . An isolated factor VII polypeptide comprising an extended recombinant polypeptide (XTEN), said XTEN comprising at least 200 amino acid residues, wherein said factor VII polypeptide exhibits a terminal half-life that is longer than about 12 hours when administered to a subject.
2 . The isolated factor VII polypeptide of claim 1 , wherein the factor VII polypeptide exhibits at least 90% sequence identity compared to a sequence selected from Table 2 when optimally aligned.
3 . The isolated factor VII polypeptide of claim 1 or 2 , wherein said factor VII is linked at its C-terminus to the XTEN.
4 . The isolated factor VII polypeptide of any of proceeding claims that is linked to the XTEN via a cleavage sequence that is cleavable by a mammalian protease selected from the group consisting of factor XIa, factor XIIa, kallikrein, factor VIIa, factor IXa, factor Xa, factor IIa (thrombin), Elastase-2, MMP13, MMP-17 and MMP-20.
5 . The isolated factor VII polypeptide of claim 4 , wherein cleavage at the cleavage sequence by the mammalian protease releases said XTEN from said factor VII polypeptide.
6 . The isolated factor VII polypeptide of any of proceeding claims, wherein said XTEN is incorporated between any two adjacent domains contained in said factor VII, wherein said two adjacent domains are selected from the group consisting of Gla, EGF1, EGF2, activated peptide (AP), and peptidase S1 (Pro).
7 . The isolated factor VII polypeptide of any of proceeding claims, wherein said XTEN is characterized in that:
(a) the cumulative total of XTEN amino acid residues is greater than 200 to about 3000 amino acid residues:
(b) the sum of asparagine and glutamine residues is less than 10% of the total amino acid sequence of the XTEN;
(c) the sum of methionine and tryptophan residues is less than 2% of the total amino acid sequence of the XTEN;
(d) the XTEN sequence has a subsequence score less than 10;
(e) the XTEN sequence has greater than 90% random coil formation as determined by GOR algorithm; and
(f) the XTEN sequence has less than 2% alpha helices and 2% beta-sheets as determined by Chou-Fasman algorithm.
8 . The isolated factor VII polypeptide of any of proceeding claims exhibiting an apparent molecular weight factor of at least about 4.
9 . The isolated factor VII polypeptide of any of proceeding claims, wherein said XTEN exhibits at least 90% sequence identity to a comparable length of an amino acid sequence selected from Table 4, Table 9, Table 10, Table 11, Table 12, or Table 13.
10 . The isolated factor VII polypeptide of any of proceeding claims that is configured according to formula VII:
(Gla)-(XTEN) u -(EGF1)-(XTEN) v -(EGF2)-(AP1)-(XTEN) w -(AP2)-(XTEN) x -(Pro)-(S) y —(XTEN) z VII
wherein independently for each occurrence,
(a) Gla is a Gla domain of factor VII;
(b) EGF1 is an EGF1 domain of factor VII;
(c) EGF2 is an EFG2 domain of factor VII;
(d) AP1 is a portion of an activator peptide domain of factor VII;
(e) AP2 is a portion of an activator peptide domain of factor IX that includes at least a first cleavage sequence;
(f) PRO is a protease domain of factor VII;
(g) S is a spacer sequence having between 1 to about 50 amino acid residues that can optionally include a cleavage sequence;
(h) XTEN is an extended recombinant polypeptide that exhibits at least 90% sequence identity to a comparable length of an amino acid sequence selected from Table 4, Table 9, Table 10, Table 11, Table 12, or Table 13,
(i) u is either 0 or 1;
(j) v is either 0 or 1;
(k) x is either 0 or 1;
(l) y is either 0 or 1; and
(m) z is either 0 or 1, with the proviso that u+v+x+y+z≧1.
11 . The isolated factor VII polypeptide of any of proceeding claims comprising more than one XTEN.
12 . An isolated factor VII polypeptide comprising at least one heterologous sequence that is cleavable by a pro-coagulant protease that does not activate a wildtype factor VII, wherein upon cleavage heterologous sequence, said factor VII polypeptide is activated.
13 . The isolated factor VII polypeptide of claim 12 that is activatable in the absence of a tissue factor.
14 . The isolated factor VII polypeptide of claim 12 or 13 , wherein the heterologous sequence is cleavable by activated factor XI.
15 . The isolated factor VII polypeptide of any of claims 12 - 14 comprising at least two heterologous sequences, each of which is cleavable by the same or different pro-coagulant proteases.
16 . The isolated factor VII polypeptide of any of claims 12 - 15 comprising an extended recombinant polypeptide (XTEN).
17 . The isolated factor VII polypeptide of any of claims 12 - 16 , wherein upon cleavage of the at least one heterologous sequence, the XTEN is also cleaved.
18 . The isolated factor VII polypeptide of any of claims 12 - 17 , wherein said heterologous sequence exhibits at least 90% sequence identity to the sequence KLTRAETVFPDVDYVNSTEAETILDNITQSTQSFNDFTRV.
19 . The isolated factor VII polypeptide of any of claims 12 - 18 , wherein said heterologous sequence exhibits at least 90% sequence identity to a sequence selected from TSKLTRAETVFP and FNDFTRV when optimally aligned.
20 . The isolated factor VII polypeptide of any of claims 12 - 19 , wherein said factor VII polypeptide does not contain a component selected the group consisting of polyethylene glycol (PEG), albumin, antibody, and an antibody fragment.
21 . The isolated factor VII polypeptide of any of claims 12 - 10 exhibiting an apparent molecular weight factor of at least about 4.
22 . A method of treating coagulopathy in a subject, comprising administering to said subject a composition comprising a therapeutically effective amount of factor VII of any of proceeding claims.
23 . The method of claim 22 , wherein said coagulopathy is hemophilia A or hemophilia B.
24 . The method of claim 22 or 23 wherein the therapeutically effective amount of factor VII is sufficient to bypass the need for exogenously administered factor VIII and/or factor IX to yield a comparable therapeutic effect.
25 . A method of treating a bleeding episode in a subject comprising: administering to said subject a composition comprising a therapeutically effective amount of the factor VII of any of claims 1 - 21 .
26 . A method of treating a subject deficient in a clotting protein, comprising: administering to said subject a composition comprising a therapeutically effective amount of the factor VII of any of claims 1 - 21 .
27 . The method of claim 26 , wherein the clotting protein substitutes wildtype factor VII, factor IX, or factor XI.