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 method for treating a bleeding disease or disorder selected from hemophilia A or hemophilia B in a subject, comprising administering to the subject an effective amount of a procoagulant compound consisting essentially of the formula:
Pep2-Zy-Bx-Pep1-Het1
wherein,
Zy is a synthetic thrombin substrate;
Bx is a self-immolative spacer;
Pep1 is a heavy chain of FVII; and
Pep2 is a light chain of FVII; and
Het1 is a half-life extending heterologous moiety.
2. The method of claim 1 , wherein the self-immolative spacer undergoes 1,4 elimination or 1,6 elimination after enzymatic cleavage of the synthetic thrombin substrate.
3. The method of claim 1 , wherein the self-immolative spacer comprises p-aminobenzyl carbamate (PABC).
4. The method of claim 1 , wherein the synthetic thrombin substrate comprises D-Phe-Pip-Arg.
5. The method of claim 1 , wherein the half-life extending heterologous moiety is albumin or an Fc region.
6. The method of claim 1 , wherein the half-life extending heterologous moiety is conjugated to Pep1 via a linker.
7. The method of 1 , wherein the half-life extending heterologous moiety is an Fc region.
8. The method of claim 1 , wherein the bleeding disorder is hemophilia A.
9. The method of claim 1 , wherein the bleeding disorder is hemophilia B.
10. The method of claim 1 , wherein the procoagulant compound consists of the formula:
Pep2-Zy-Bx-Pep1-Het1
wherein,
Zy is a synthetic thrombin substrate;
Bx is a self-immolative spacer;
Pep1 is a heavy chain of FVII;
Pep2 is a light chain of FVII; and
Het1 is a half-life extending heterologous moiety.
11. The method of claim 1 , wherein the half-life extending heterologous moiety is a low complexity polypeptide, albumin, albumin binding polypeptide or fatty acid, Fc, transferrin, a PAS sequence, the C-terminal peptide (CTP) of the β subunit of human chorionic gonadotropin, polyethylene glycol (PEG), hydroxyethyl starch (HES), albumin-binding small molecules, vWF, a clearance receptor or fragment thereof which blocks binding of the procoagulant compound to a clearance receptor, or any combination thereof.
12. The method of claim 10 , wherein the half-life extending heterologous moiety is a low complexity polypeptide, albumin, albumin binding polypeptide or fatty acid, Fc, transferrin, a PAS sequence, the C-terminal peptide (CTP) of the β subunit of human chorionic gonadotropin, polyethylene glycol (PEG), hydroxyethyl starch (HES), albumin-binding small molecules, vWF, a clearance receptor or fragment thereof which blocks binding of the procoagulant compound to a clearance receptor, or any combination thereof.