Methods and compositions for treating bleeding disorders
View Patent ↗Aspects of the invention include methods for enhancing blood coagulation in a subject. In practicing methods according to certain embodiments, an amount of a non-anticoagulant sulfated polysaccharide (NASP) is administered to a subject to enhance blood coagulation in the subject. Also provided are methods for preparing a NASP composition having blood coagulation enhancing activity. Compositions and kits for practicing methods of the invention are also described.
1. A method of identifying a tissue factor pathway inhibitor (TFPI)-inhibiting compound which is capable of enhancing blood coagulation, the method comprising:
contacting a first portion of a plasma sample comprising TFPI with an antibody which inhibits TFPI activity;
measuring peak thrombin generation or peak time of the first portion of plasma sample;
contacting a second portion of the plasma sample comprising TFPI with a composition comprising a sulfated polysaccharide having a fucose content of 60% or greater;
measuring peak thrombin generation or peak time in the second portion of plasma sample; and
comparing the peak thrombin generation or peak time of the second portion of plasma sample with the peak thrombin generation or peak time of the first portion of plasma sample, wherein no difference in peak thrombin generation or peak time between the second portion of plasma sample compared to the first portion of plasma sample indicates that the sulfated polysaccharide is a TFPI-inhibiting compound.
2. The method according to claim 1 , wherein the antibody is an anti-TFPI C-terminus antibody.
3. The method according to claim 1 , wherein the plasma sample are normal is Factor VIII inhibited plasma.
4. The method according to claim 1 , wherein the sulfated polysaccharide is a fucoidan.
5. The method according to claim 1 , wherein the fucoidan is selected from the group consisting of Fucoidan 5307002, Fucus vesiculosus ; Fucoidan L/FVF1091, Fucus vesiculosus ; Fucoidan DS100159, Fucus vesiculosus ; Fucoidan DS100160, Fucus vesiculosus ; and Fucoidan VG2010100, Fucus vesiculosus .
6. The method according to claim 1 , wherein the composition comprising the sulfated polysaccharide comprises an alginate content of 10% or less.
7. The method according to claim 1 , wherein the sulfated polysaccharide is Fucoidan L/FVF1091.
8. The method according to claim 1 , wherein the antibody is an antibody directed against the C-terminus of TFPI.
9. The method according to claim 1 , wherein the TFPI in the first portion of plasma and the second portion of plasma is full-length TFPI.
10. The method according to claim 9 , further comprising contacting the first portion of plasma with recombinant C-terminus truncated TFPI (TFPI 1-160).
11. The method according to claim 10 , wherein an increase in peak thrombin generation or decrease in peak time of the second portion of plasma compared to the first portion of plasma indicates that the sulfated polysaccharide is a C-terminal TFPI-binding compound.
12. A method of identifying a C-terminal TFPI-binding compound which is capable of enhancing blood coagulation, the method comprising:
contacting a first portion of a plasma sample comprising full-length TFPI and an antibody directed against the C-terminus of TFPI with recombinant C-terminus truncated TFPI (TFPI 1-160);
measuring a first peak thrombin generation or first peak time of the first portion of the plasma sample after contacting with the recombinant C-terminus truncated TFPI (TFPI 1-160);
contacting a second portion of the plasma sample with a composition comprising a sulfated polysaccharide having a fucose content of 60% or greater;
measuring a second peak thrombin generation or second peak time after contacting the second portion of the plasma sample with the composition comprising the sulfated polysaccharide having a fucose content of 60% or greater; and
comparing the measured first peak thrombin generation or the measured first peak time with the measured second peak thrombin generation or the measured second peak time, wherein an increase in peak thrombin generation or decrease in peak time of the second portion of the plasma sample indicates that the test compound is a C-terminal TFPI-binding compound.
13. The method according to claim 12 , wherein the fucoidan is selected from the group consisting of Fucoidan 5307002, Fucus vesiculosus ; Fucoidan L/FVF1091, Fucus vesiculosus ; Fucoidan DS100159, Fucus vesiculosus ; Fucoidan DS100160, Fucus vesiculosus ; and Fucoidan VG2010100, Fucus vesiculosus .
14. The method according to claim 12 , wherein the sulfated polysaccharide is Fucoidan L/FVF1091.