Regulation of von Willebrand factor (VWF)
The present disclosure relates to agents, compositions targeting to von Willebrand factor (VWF). The VWF targeting agents are synthetic polynucleotides, including VWF binding agents and their reversal agents. The VWF binding agents are VWF binding aptamers that bind to and inhibit the VWF activities. The VWF binding agents can be reversed using reversal agents to reverse the inhibitory effect and thereby restore VWF activities. The disclosure further provides methods for regulating the activities of VWF, thereby modulating VWF mediated platelet functionality, such as thrombosis, the present VWF targeting agents may be used for preventing thrombus formation and treating thrombotic disorders.
1 . A method for treating a clinical condition associated with elevated levels of plasma VWF in a patient comprising: administrating to the patient a VWF binding aptamer comprising the nucleic acid sequence presented by SEQ ID No: 3; and reversing the effect of the VWF binding aptamer using a reversal agent comprising the nucleic acid sequence presented by SEQ ID No: 9, wherein the reversal agent is administered at a ratio of 1:1 relative to the VWF binding aptamer or less in moles when the levels of plasma VWF need to be increased in the patient receiving the treatment of the VWF binding aptamer.
2 . The method of claim 1 , wherein the clinical condition associated with elevated levels of plasma VWF comprises systemic lupus erythematosus (SLE), first ischemic stroke, secondary stroke, transient ischemic attack (TIA), silent stroke, a cardiovascular disease, diabetic disease, and cancer metastasis.
3 . The method of claim 1 , wherein the VWF binding aptamer includes at least one nucleotide modification with 2′-O-methyl modification, and wherein the reversal agent includes at least one nucleotide modification with 2′-O-methyl modification.
4 . The method of claim 3 , wherein the VWF binding aptamer is modified with a conjugate selecting from the group consisting of a PEG polymer, a protein, an antibody or variant thereof, a peptide, a lipid, a fatty acid, a carbohydrate, and a small molecule, and/or wherein the reversal agent is modified with a conjugate selecting from the group consisting of a PEG polymer, a protein, an antibody or variant thereof, a peptide, a lipid, a fatty acid, a carbohydrate, and a small molecule.
5 . The method of claim 1 , wherein the VWF binding aptamer comprises a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 4-6, and wherein the reversal agent comprises the nucleic acid sequence of SEQ ID No: 10.
6 . The method of claim 1 wherein the ratio of the reversal agent and the VWF binding aptamer is, at 1:1.5, at 1:2, at 1:3, at 1:4, or at 1:5 in moles.
7 . The method of claim 1 , wherein the amount of the reversal agent is administered to reverse the activity of the VWF binding aptamer by 50 to 100%.
8 . The method of claim 1 , wherein the reversal agent is administered when the patient receiving the treatment of the VWF binding aptamer is under a risk of excessive bleeding.
9 . The method of claim 1 , wherein the reversal agent is administered when the patient receiving the treatment of the VWF binding aptamer is receiving a clinical surgery.
10 . A method of modulating VWF activity in a subject comprising administering to the circulatory system of the subject a VWF binding aptamer having the nucleic acid sequence presented by SEQ ID NO: 3, and reversing the effect of the VWF binding aptamer using a reversal agent having the nucleic acid sequence presented by SEQ ID NO: 9 wherein the reversal agent is used at a ratio of 1:1 to the VWF binding aptamer or less in moles when the activity of plasma VWF needs to be increased.
11 . The method of claim 10 , wherein the VWF binding aptamer includes at least one nucleotide modification with 2′-O-methyl modification, and wherein the reversal agent includes at least one nucleotide modification with 2′-O-methyl modification.
12 . The method of claim 11 , wherein the VWF binding aptamer is modified with a conjugate selecting from the group consisting of a PEG polymer, a protein, an antibody or variant thereof, a peptide, a lipid, a fatty acid, a carbohydrate, and a small molecule, and/or wherein the reversal agent is modified with a conjugate selecting from the group consisting of a PEG polymer, a protein, an antibody or variant thereof, a peptide, a lipid, a fatty acid, a carbohydrate, and a small molecule.
13 . The method of claim 11 , wherein the VWF binding aptamer comprises a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 4-6, and wherein the reversal agent comprises the nucleic acid sequence of SEQ ID No: 10.
14 . The method of claim 10 , wherein the ratio of the reversal agent and the VWF binding aptamer is, at 1:1.5, at 1:2, at 1:3, at 1:4, or at 1:5 in moles.
15 . The method of claim 10 , wherein the amount of the reversal agent is used to reverse the activity of the VWF binding aptamer by 50 to 100%.