Evacuated blood collection tubes containing protease inhibitors for the assessment of contact system activation

Patent metadata

Field	Value
Publication number	US-11458477-B2
Application number	US-202017102509-A
Country	US
Kind code	B2
Filing date	Nov 24, 2020
Priority date	Aug 13, 2015
Publication date	Oct 4, 2022
Grant date	Oct 4, 2022

Abstract

Disclosed herein are evacuated blood collection tubes comprising protease inhibitor cocktails in liquid form and uses thereof for assessing features associated with the contact system in a subject, including the endogenous level of contact system activation, the endogenous level of a drug that targets a component of contact system during treatment, and/or the immunogenicity of such a drug.

First claim

What is claimed is:

1. A method for assessing the level of a drug targeting the contact system in a subject, the method comprising:
(i) collecting blood from a subject to an evacuated blood collection tube; wherein the subject has been administered a drug that targets a component of the contact system;
(ii) processing the blood to produce a plasma sample; and
(iii) measuring the level of contact system activation in the plasma sample, wherein the evacuated blood collection tube comprises a liquid formulation that comprises a mixture of protease inhibitors, polybrene, and EDTA, wherein the mixture of protease inhibitors comprises benzamidine, soybean trypsin inhibitor, leupeptin, and 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF);
wherein the drug is a plasma kallikrein inhibitor.

2. The method of claim 1 , wherein the tube is a non-glass tube.

3. The method of claim 1 , wherein the liquid formulation comprises 80-120 mM benzamidine, 1-3 mg/ml soybean trypsin inhibitor, 200-300 μM leupeptin, and 10-30 mM AEBSF.

4. The method of claim 1 , wherein the liquid formulation comprises 100 mM benzamidine, 2 mg/ml soybean trypsin inhibitor, 263 μM leupeptin, and 20 mM AEBSF.

5. The method of claim 1 , wherein the polybrene is at a concentration of 400 μg/mL and the EDTA is at a concentration of 20 mM.

6. The method of claim 1 , wherein the liquid formulation further comprises trisodium citrate, citric acid, and dextrose, optionally wherein the trisodium citrate is at a concentration of 100 mM, the citric acid is at a concentration of 67 mM, and the dextrose is 2%.

7. The method of claim 1 , wherein the liquid formulation has a pH of 4-6, optionally wherein the liquid formulation has a pH of 4.5.

8. The method of claim 1 , wherein in step (i), the evacuated blood collection tube is not the first tube filled with the blood from the subject; and/or step (ii) is performed within one hour after step (i).

9. The method of claim 1 , wherein the subject is a human subject having a disease associated with the contact system, optionally wherein the disease is hereditary angioedema (HAE) or idiopathic angioedema.

10. The method of claim 1 , wherein plasma kallikrein inhibitor is an antibody that binds active plasma kallikrein.

11. A method for assessing immunogenicity of a drug targeting the contact system, the method comprising:
(i) collecting blood from a subject to an evacuated blood collection tube; wherein the subject has been administered a drug that targets a component of the contact system;
(ii) processing the blood to produce a plasma sample; and
(iii) measuring the level of antibodies that bind to the drug in the plasma sample, wherein the evacuated blood collection tube comprises a liquid formulation that comprises a mixture of protease inhibitors, polybrene, and EDTA, wherein the mixture of protease inhibitors comprises benzamidine, soybean trypsin inhibitor, leupeptin, and 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF);
wherein the drug is a plasma kallikrein inhibitor.

12. The method of claim 11 , wherein the liquid formulation comprises 80-120 mM benzamidine, 1-3 mg/ml soybean trypsin inhibitor, 200-300 μM leupeptin, and 10-30 mM AEBSF.

13. The method of claim 11 , wherein the liquid formulation comprises 100 mM benzamidine, 2 mg/ml soybean trypsin inhibitor, 263 μM leupeptin, and 20 mM AEBSF.

14. The method of claim 11 , wherein the polybrene is at a concentration of 400 μg/mL and the EDTA is at a concentration of 20 mM.

15. The method of claim 11 , wherein the liquid formulation further comprises trisodium citrate, citric acid, and dextrose, optionally wherein the trisodium citrate is at a concentration of 100 mM, the citric acid is at a concentration of 67 mM, and the dextrose is 2%.

16. The method of claim 11 , wherein the liquid formulation has a pH of 4-6, optionally wherein the liquid formulation has a pH of 4.5.

17. The method of claim 11 , wherein in step (i), the evacuated blood collection tube is not the first tube filled with the blood from the subject; and/or step (ii) is performed within one hour after step (i).

18. The method of claim 11 , wherein the subject is a human subject having a disease associated with the contact system, optionally wherein the disease is hereditary angioedema (HAE) or idiopathic angioedema.

19. The method of claim 11 , further comprises, prior to step (iii), isolating antibodies that bind the drug from the plasma sample.

20. The method of claim 11 , wherein plasma kallikrein inhibitor is an antibody that binds active plasma kallikrein.

Assignees

• Takeda Pharmaceuticals Co

Inventors

• Sexton Daniel J
• Faucette Ryan

Classifications

• B01L3/5082 Primary

  Test tubes per se · CPC title

• A61B5/154 Primary

  {Devices} using pre-evacuated means · CPC title

• C07K16/40

  against enzymes · CPC title

• B01L2300/0832

  cylindrical, tube shaped · CPC title

• G01N33/564

  for pre-existing immune complex or autoimmune disease {, i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9} · CPC title