Blood factor monitoring assay and uses thereof

• US 10,001,495 B2
• Filed: 07/25/2013
• Issued: 06/19/2018
• Est. Priority Date: 07/25/2012
• Status: Active Grant

• Current Assignee: Bioverativ Therapeutics Inc. (Sanofi )
• Original Assignee: Bioverativ Therapeutics Inc. (Sanofi )
• Inventors: Sommer, Jurg
• Primary Examiner(s): SZPERKA, MICHAEL EDWARD
• Application Number:
  US14/416,214
• Publication Number:
  US 20150185235A1
• Time in Patent Office: 1,790 Days
• Field of Search: None
• CPC Class Codes: A61K 38/4846 Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38)
• CPC Class Codes: C12Y 304/21022 Coagulation factor IXa (3.4.21.22)
• CPC Class Codes: G01N 2333/755 Factors VIII, e.g. factor VIII C [AHF], factor VIII Ag [VWF]
• CPC Class Codes: G01N 2333/9645 Factor IX (3.4.21.22)
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 2800/224 Haemostasis or coagulation
• CPC Class Codes: G01N 33/86 involving blood coagulating time or factors, or their receptors
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743

Abstract

The present disclosure provides methods and compositions for diagnosing and treating subject having a bleeding disorder. The disclosed methods comprise contacting a sample, e.g., a blood or plasma sample obtained from the patient, with an activation mixture comprising an activated coagulation factor and a phospholipid mixture, wherein the activation mixture is dried onto a solid substrate. Also provided is a global hemostasis test based on the integration of clotting time (Ct) and pharmacokinetics data. The methods and compositions presented can be applied to point-of-care diagnostic systems.

Claims

1. A method of treating a bleeding disorder in a patient in need thereof comprising:

a) measuring the time between contacting of a sample obtained from the patient with an activation mixture dried on a solid substrate and the onset of clotting, thereby calculating the clotting time (Ct),

wherein the activation mixture comprises (i) an activated Factor IX (FIXa) and (ii) a phospholipid mixture,

wherein the Ct is used to determine a pharmacokinetic (PK) parameter of a coagulation factor, wherein the coagulation factor comprises a Factor VIII (FVIII),

wherein the PK parameter is a terminal half-life (HL) or a time to trough (T),

wherein the HL is calculated according to the following formula;

HL=−

0.693×

(T₂−

T₁)×

A/(Ct₁−

Ct₂),

wherein A is a constant value corresponding to the slope of a Ct versus coagulation factor concentration dose-response, T₁ and T₂ are times at which Ct is measured, and Ct₁ and Ct₂ are Ct values measured at T₁ and T₂, respectively,

wherein the T is calculated according to the following formula;

T=−

1.44×

HL/(A×

(Ct_measured−

Ct_trough),

wherein A is a constant value corresponding to the slope of a Ct versus coagulation factor concentration dose-response, and HL is the terminal half-life, Ct_measured is Ct measured at certain time point, and Ct_trough is patient-specific clot time at trough, and

b) administering an effective amount of the coagulation factor at a dosing interval based on the PK parameter.

3. The method of claim 1, wherein the correlation between Ct and a coagulation factor level (% Factor) is calculated based on the following formula:

Ct=A×

Ln(% Factor)+B

wherein, for each coagulation factor, A is a constant value corresponding to the slope of a Ct versus coagulation factor concentration dose-response, and B is a patient-specific off-set value.

4. The method of claim 1, further comprising monitoring the efficacy of the coagulation factor administered to the patient.

5. The method of claim 1, wherein the PK parameter is terminal half-life (HL).

6. The method of claim 1, wherein the PK parameter is time to trough (T).

7. The method of claim 6, wherein the patient is administered a new dose of the coagulation factor every T interval.

8. The method of claim 1, wherein the sample is selected from the group consisting of whole blood, citrated or equivalently stabilized blood, plasma, and other fluid sample containing or suspected of containing a coagulation factor.

9. The method of claim 8, wherein the sample further comprises an added inhibitor.

10. The method of claim 1, wherein the Ct correlates with a therapeutically efficacious treatment.

11. The method of claim 1, wherein the FVIII is a chimeric FVIII polypeptide comprising FVIII and a non-FVIII polypeptide, wherein the non-FVIII polypeptide increases a half-life of the FVIII and comprises Fc, albumin, a PAS sequence, transferrin, C-terminal peptide of hCG (CTP), polyethylene glycol (PEG), hydryoxyethyl starch (HES), or two or more combinations thereof.

13. The method of claim 11, wherein the chimeric FVIII polypeptide comprises a monomer dimer hybrid comprising the FVIII and an Fc.

2. A method of treating a bleeding disorder in a patient in need thereof comprising:

a) measuring the time between contacting of a sample obtained from the patient with an activation mixture dried on a solid substrate and the onset of clotting, thereby calculating the clotting time (Ct),

wherein the activation mixture comprises (i) an activated Factor XI (FXIa) and (ii) a phospholipid mixture,

wherein the Ct is used to determine a pharmacokinetic (PK) parameter of a coagulation factor, wherein the coagulation factor comprises a Factor IX (FIX),

wherein the PK parameter is a terminal half-life (HL) or a time to trough (T),

wherein the HL is calculated according to the following formula;

HL=−

0.693×

(T₂−

T₁)×

A/(Ct₁−

Ct₂),

wherein, for each coagulation factor, A is a constant value corresponding to the slope of a Ct versus coagulation factor concentration dose-response, T₁ and T₂ are times at which Ct is measured, and Ct₁ and Ct₂ are Ct values measured at T₁ and T₂, respectively,

wherein the T is calculated according to the following formula;

T=−

1.44×

HL/(A×

(Ct_measured−

Ct_trough),

wherein for each coagulation factor, A is a constant value corresponding to the slope of a Ct versus coagulation factor concentration dose-response, and HL is the terminal half-life, Ct_measured is Ct measured at certain time point, and Ct_trough is patient-specific clot time at trough, and

b) administering an effective amount of the coagulation factor at a dosing interval based on the PK parameter.

12. The method of claim 2, wherein the FIX is a chimeric FIX polypeptide comprising FIX and a non-FIX polypeptide, wherein the non-FIX polypeptide increases a half-life of the FIX and comprises Fc, albumin, a PAS sequence, transferrin, C-terminal peptide of hCG (CTP), polyethylene glycol (PEG), hydryoxyethyl starch (HES), or two or more combination thereof.

14. The method of claim 12, wherein the chimeric FIX polypeptide comprises a monomer dimer hybrid comprising the FIX and an Fc.

15. The method of claim 2, wherein the correlation between Ct and a coagulation factor level (% Factor) is calculated based on the following formula:

Ct=A×

Ln(% Factor)+B,

wherein A is a constant value corresponding to the slope of a Ct versus coagulation factor concentration dose-response, and B is a patient-specific off-set value.

16. The method of claim 2, further comprising monitoring the efficacy of the coagulation factor administered to the patient.

17. The method of claim 2, wherein the PK parameter is terminal half-life (HL).

18. The method of claim 2, wherein the PK parameter is time to trough (T).

19. The method of claim 18, wherein the patient is administered a new dose of the coagulation factor every T interval.

20. The method of claim 2, wherein the sample is selected from the group consisting of whole blood, citrated or equivalently stabilized blood, plasma, and other fluid sample containing or suspected of containing a coagulation factor.

21. The method of claim 20, wherein the sample further comprises an added inhibitor.

22. The method of claim 2, wherein the Ct correlates with a therapeutically efficacious treatment.