Von willebrand factor- and factor VIII-polymer conjugates having a releasable linkage
The present invention provides von Willebrand Factor-polymer conjugates and Factor VIII-polymer conjugates, each having a releasable linkage. Methods of making conjugates, methods for administering conjugates, are also provided.
1. A method of treating a bleeding disorder in a subject, said method comprising administering a therapeutically effective amount of a modified protein having an extended in-vivo half life over the unmodified parent protein, wherein the unmodified parent protein is either a von Willebrand Factor moiety or a Factor VIII moiety and the modified protein is a conjugate having a formula:
wherein:
POLY 1 is a first water-soluble polymer;
POLY 2 is a second water-soluble polymer;
X 1 is a first spacer moiety;
X 2 is a second spacer moiety;
H α is an ionizable hydrogen atom;
R 1 is H or an organic radical;
R 2 is H or an organic radical;
(a) is either zero or one;
(b) is either zero or one;
R e1 , when present, is a first electron altering group;
R e2 , when present, is a second electron altering group;
Y 1 is O or S; and
Y 2 is O or S, such that
from the formula is a releasable linkage that is selected from a carbamate linkage, a thiocarbamate linkage, and a dithiocarbamate linkage; and
(vWF/F8) is a residue of an amine-containing biologically active agent that is either a von Willebrand Factor moiety or a Factor VIII moiety.
2. The method of claim 1 , wherein
from the formula is a releasable linkage that is either a carbamate linkage or a thiocarbamate linkage.
3. The method of claim 2 , wherein the releasable linkage is a carbamate linkage, and the compound possesses the structure:
4. The method of claim 1 , wherein the first water-soluble polymer is a poly(alkylene oxide) and the second water-soluble polymer is a poly(alkylene oxide).
5. The method of claim 4 , wherein the first water-soluble polymer has a weight-average molecular weight of between 10,000 Daltons to 85,000 Daltons and the second water-soluble polymer has a weight-average molecular weight of between 10,000 Daltons to 85,000 Daltons.
6. The method of claim 1 , wherein the amine-containing biologically active agent is a von Willebrand Factor moiety.
7. The method of claim 6 , wherein the von Willebrand Factor moiety is human recombinant von Willebrand Factor.
8. The method of claim 1 , wherein the amine-containing biologically active agent is a Factor VIII moiety.
9. The method of claim 8 , wherein the Factor VIII moiety is human recombinant B-domain deleted Factor VIII.
10. The method of claim 8 , wherein the Factor VIII moiety is human recombinant full length Factor VIII.
11. The method of claim 1 , wherein the conjugate is comprised in a composition together with one or more pharmaceutically acceptable excipients.
12. The method of claim 11 , wherein said administering is by injection.
13. The method of claim 12 , wherein the composition is in a form selected from the group consisting of a powder that can be reconstituted, a suspension, and a solution.
14. The method of claim 12 , wherein the composition is in a unit dosage form.
15. The method of claim 1 , wherein the modified protein has a structure selected from the group consisting of:
wherein, for each structure and in each instance, (n) is independently an integer from 4 to 1500.
16. The method of claim 15 , wherein the amine-containing biologically active agent is a von Willebrand Factor moiety.
17. The method of claim 16 , wherein the von Willebrand Factor moiety is human recombinant von Willebrand Factor.
18. The method of claim 15 , wherein the amine-containing biologically active agent is a Factor VIII moiety.
19. The method of claim 17 , wherein the Factor VIII moiety is human recombinant B-domain deleted Factor VIII.
20. The method of claim 15 , wherein the Factor VIII moiety is human recombinant full length Factor VIII.