Subcutaneous administration of modified factor IX polypeptides and treatment of hemophilia B
Methods for prophylactic treatment of hemophilia B are provided in which modified FIX polypeptides, particularly FIXa, are subcutaneously administered daily or less frequently, such as every other day, or every 2, 3 or 4 days. The treatment results in normal coagulation pharmacokinetics and normal levels of FIX, or mild hemophilia B.
1. A method of prophylactically treating hemophilia B, comprising a regimen of subcutaneously administering, to a subject with hemophilia B, a modified Factor IX (FIX) polypeptide daily or every 2 or every 3 or every 4 days, wherein:
treatment results in normal or near normal levels of FIX to effect normal coagulation or mild hemophilia;
the modified FIX polypeptide has coagulation activity of at least about 7-10 times the wild-type FIX of SEQ ID NO: 3 or 20, and the modified FIX polypeptide has increased serum half-life when it is administered subcutaneously compared to when it is administered intravenously, whereby levels of FIX are maintained by the daily, every 2-day, every 3-day or every 4-day regimen and dose;
each dose is about or is 40-400 IU of modified FIX/kg of the subject;
normal or near normal coagulation is FIX activity of about 40%-150% of the activity in blood, relative to the WHO 4th International Standard, where 100 IU/dL is 100% FIX activity;
and mild hemophilia coagulation is FIX activity of about 5% to 40% of the activity in blood, relative to the WHO 4th International Standard, where 100 IU/dL is 100% FIX activity;
wherein the modified FIX polypeptide comprises replacements corresponding to R318Y/R338E/T343R.
2. The method of claim 1 , comprising administering a sufficient number of doses to achieve a level of FIX activity that is >20%, wherein:
100 IU/dL is 100% FIX activity; and
the number of doses is sufficient to saturate the extracellular compartment and reach a steady-state level of FIX activity >20% and to remain at the level as long as the dosage and regimen are implemented.
3. The method of claim 2 , wherein the number of doses to achieve the level is 5, 6, or 7.
4. The method of claim 1 , wherein each dose, which is administered daily or every other day, is about 40-400 IU of modified FIX/kg of the subject.
5. The method of claim 1 , comprising administering an intravenous dose or doses to saturate the extravascular space prior to commencing the subcutaneous regimen.
6. The method of claim 1 , wherein a single intravenous dose of 25-250 IU/kg or 50-150 IU/kg of the modified FIX polypeptide is administered prior to the subcutaneous regimen.
7. The method of claim 1 , wherein:
the subcutaneous regimen is selected from:
a daily dose of 50-100 IU/kg; a daily dose of 40-350 IU/kg or 40-300 IU/kg; a daily dose of 70-150 IU/kg; a daily dose of 80-120 IU/kg or 81-118 IU/kg or a daily dose of 40-120 IU/kg; or a daily dose of 40-150 IU/kg, or a daily dose of 50-175 IU/kg, or a daily dose of 80-300 IU/kg, or a daily dose of 75-200 IU/kg, or a daily dose of 80-150 IU/kg.
8. The method of claim 1 , wherein the modified FIX polypeptide also has increased serum half-life compared to a wild-type FIX of SEQ ID NO: 3 or 20.
9. The method of claim 1 , wherein the modified FIX polypeptide has at least greater than 10-fold, greater than 15-fold or greater than 20-fold activity than the wild-type FIX of SEQ ID NO: 3 or 20.
10. The method of claim 8 , wherein the modified FIX polypeptide is further modified by PEGylation, albumination, glycosylation, farnesylation, carboxylation, hydroxylation, phosphorylation, and/or multimerization conjugation.
11. The method of claim 10 , wherein the modified FIX polypeptide is albuminated, hyperglycosylated, PEGylated, or linked to an Fc multimerization domain.
12. The method of claim 11 , wherein the modified FIX polypeptide comprises replacements corresponding to R318Y/R338E/T343R.
13. The method of claim 12 , wherein the modified FIX polypeptide comprises the sequence of amino acids set forth in SEQ ID NO:394.
14. The method of claim 1 , wherein:
the modified FIX polypeptide comprises amino acid replacements selected from among replacements: R318Y/R338E/T343R/R403E/E410N, R318Y/R338E/T343R/E410N, Y155F/R318Y/R338E/T343R/R403E, Y155F/K228N/K247N/N249S/R318Y/R338E/T343R/R403E/E410N, Y155F/K247N/N249S/R318Y/R338E/T343R/R403E, K247N/N249S/R318Y/R338E/T343R/R403E, R318Y/R338E/T343R, Y155F/K247N/N249S/R318Y/R338E/T343R, K228N/R318Y/R338E/T343R/R403E/E410N, K228N/K247N/N249S/R318Y/R338E/T343R/R403E, R318Y/R338E/T343R/R403E/E410S, K247N/N249S/R318Y/R338E/T343R, Y155F/R318Y/R338E/T343R/R403E/E410N, D104N/K106S/R318Y/R338E/T343R/R403E/E410N, R318Y/R338E/T343R/N346Y/R403E/E410N, R318Y/R338E/T343R/N346D/R403E/E410N, R318Y/R338E/T343R/R403E, Y155F/R318Y/R338E/T343R/E410N Y155F/K247N/N249S/R318Y/R338E/T343R/R403E/E410N, N260S/R318Y/R338E/T343R/R403E/E410N, Y155F/N260S/R318Y/R338E/T343R/R403E/E410N, K228N/K247N/N249S/R318Y/R338E/T343R/R403E/E410N, K247N/N249S/R318Y/R338E/T343R/E410N, Y155F/K247N/N249S/R318Y/R338E/T343R/E410N, Y155F/K247N/N249S/R318Y/T343R/R403E/E410N, Y155F/K247N/N249S/R318Y/T343R/R403E, K247N/N249S/R318Y/T343R/R403E, K247N/N249S/R318Y/T343R/E410N, Y155F/R318Y/R338E/T343R, K228N/K247N/N249S/R318Y/R338E/T343R/E410N, and K228N/K247N/N249S/R318Y/T343R/R403E/E410N, or the same replacements at corresponding amino acid residues in an unmodified FIX polypeptide; and
the unmodified FIX polypeptide comprises the sequence of amino acids set forth in SEQ ID NO: 3 or 20.
15. The method of claim 14 , wherein the modified FIX polypeptide comprises amino acid replacements selected from among replacements R318Y/R338E/T343R/R403E/E410N, R318Y/R338E/T343R/E410N, Y155F/R318Y/R338E/T343R/R403E, Y155F/K228N/K247N/N249S/R318Y/R338E/T343R/R403E/E410N, Y155F/K247N/N249S/R318Y/R338E/T343R/R403E, K247N/N249S/R318Y/R338E/T343R/R403E, R318Y/R338E/T343R, Y155F/K247N/N249S/R318Y/R338E/T343R, K228N/R318Y/R338E/T343R/R403E/E410N, K228N/K247N/N249S/R318Y/R338E/T343R/R403E, R318Y/R338E/T343R/R403E/E410S, and K247N/N249S/R318Y/R338E/T343R.
16. The method of claim 14 , wherein the modified FIX polypeptide comprises amino acid replacements R318Y/R338E/T343R/R403E/E410N or R318Y/R338E/T343R/E410N.
17. The method of claim 1 , wherein the modified FIX polypeptide is in a pharmaceutical composition comprising the modified FIX polypeptide in a pharmaceutically acceptable vehicle formulated for subcutaneous administration.
18. The method of claim 17 , wherein the composition is formulated so that a single dose of the composition is less than 1 ml, 1 ml to 10 ml, or 0.5 ml to 10 ml, or 0.5 ml to 5 ml, or 0.5 ml to 3 ml; and
each range is inclusive.
19. The method of claim 1 , wherein the subject treated is a neonate, an infant, a toddler, or a child of 3 years to 18 years.
20. The method of claim 1 , wherein the FIX polypeptide that is administered is the active form of FIX (FIXa).
21. The method of claim 1 , wherein the subcutaneous regimen comprises alternate day dosing of about 260-300 IU/kg.
22. A method of prophylactically treating hemophilia B, comprising a regimen of subcutaneously administering, to a subject with hemophilia B, a modified Factor IX (FIX) polypeptide daily or every 2 or every 3 or every 4 days, wherein:
treatment results in normal or near normal levels of FIX to effect normal coagulation or mild hemophilia;
the modified FIX polypeptide has coagulation activity of at least about 7-10 times the wild-type FIX of SEQ ID NO: 3 or 20, and the modified FIX polypeptide has increased serum half-life when it is administered subcutaneously compared to when it is administered intravenously, whereby levels of FIX are maintained by every 3-day regimen and dose;
each dose is about or is 480-560 IU of modified FIX/kg of the subject;
normal or near normal coagulation is FIX activity of about 40%-150% of the activity in blood, relative to the WHO 4th International Standard, where 100 IU/dL is 100% FIX activity;
mild hemophilia coagulation is FIX activity of about 5% to 40% of the activity in blood, relative to the WHO 4th International Standard, where 100 IU/dL is 100% FIX activity; and
the modified FIX polypeptide comprises replacements corresponding to R318Y/R338E/T343R.
23. The method of claim 1 , wherein the treatment effects normal or near normal coagulation.
24. The method of claim 1 , wherein the modified FIX comprises the sequence of amino acids set forth in SEQ ID NO:394.
25. The method of claim 1 , wherein, after a pre-determined number of doses sufficient to saturate the extracellular space, the level of FIX is >30%, where 100 IU/dL is 100% FIX activity, and remains at the level as long as the dosage and regimen are implemented.
26. The method of claim 21 , wherein the treatment effects normal or near normal coagulation.
27. The method of claim 17 , wherein the composition is formulated so that a single dose of the composition is less than 1 ml.