Procoagulant compounds
The present disclosure provides protease-activatable procoagulant compounds comprising a procoagulant polypeptide, e.g., a procoagulant peptide and/or clotting factor, and a linker comprising a protease-cleavable substrate (e.g., a synthetic thrombin substrate) and a self-immolative spacer (e.g., p-amino benzyl carbamate). Upon cleavage of the protease-cleavable substrate by a protease (e.g., thrombin), the self-immolative spacer cleaves itself from the procoagulant polypeptide such that the polypeptide is in an underivatized and active form. Also provided are pharmaceutical compositions, methods for treating bleeding disorders using the disclosed compounds, methods of enhancing in vivo efficacy of procoagulant polypeptides, methods of increasing the efficacy of proteolytic cleavage of compounds comprising procoagulant polypeptides, methods of activating procoagulant polypeptides, and methods of releasing a procoagulant polypeptide from a heterologous moiety such as PEG.
1. A procoagulant compound comprising a formula Pep2-Zy-Bx-Pep1, wherein
Pep2 is a light chain of FVII;
Zy is a synthetic thrombin substrate;
Bx is a self-immolative spacer; and
Pep1 is a heavy chain of FVII.
2. The procoagulant compound according to claim 1 , wherein the self-immolative spacer undergoes 1,6 elimination after the enzymatic cleavage of the synthetic thrombin substrate.
3. The procoagulant compound according to claim 1 , wherein the self-immolative spacer is p-aminobenzyl carbamate (PABC).
4. The procoagulant compound according to claim 1 , wherein the synthetic thrombin substrate comprises the sequence D-Phe-Pip-Arg.
5. The procoagulant compound according to claim 1 , further comprising at least one heterologous moiety that extends the half-life of the procoagulant compound, wherein the at least one heterologous moiety is conjugated to Pep1.
6. The procoagulant compound according to claim 5 , wherein the at least one heterologous moiety is conjugated to Pep1 via a linker.
7. The procoagulant compound according to claim 5 , wherein the at least one heterologous moiety comprises an Fc region.
8. The procoagulant compound according to claim 5 , comprising two heterologous moieties that extend the half-life of the procoagulant compound.
9. The procoagulant compound according to claim 8 , wherein the two heterologous moieties are joined by a linker.
10. The procoagulant compound according to claim 9 , wherein each of the two heterologous moieties comprises an Fc region.
11. A pharmaceutical composition comprising the procoagulant compound according to claim 1 , and a pharmaceutically acceptable carrier.