Methods and compositions for treating hemophilia B

• US 9,629,930 B2
• Filed: 10/08/2015
• Issued: 04/25/2017
• Est. Priority Date: 10/12/2010
• Status: Active Grant

• Current Assignee: Children's Hospital of Philadelphia (Children's Hospital of Philadelphia Foundation), Sangamo Biosciences, Inc.
• Original Assignee: Children's Hospital of Philadelphia (Children's Hospital of Philadelphia Foundation), Sangamo Biosciences, Inc.
• Inventors: Gregory, Philip D., High, Katherine A., Holmes, Michael C., Li, Hojun
• Primary Examiner(s): Babic, Christopher M
• Assistant Examiner(s): Hiriyanna, Kelaginamane T
• Application Number:
  US14/878,703
• Publication Number:
  US 20160022838A1
• Time in Patent Office: 565 Days
• Field of Search: 424 931
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61N 2005/0661 ultraviolet
• CPC Class Codes: A61N 5/10 X-ray therapy; Gamma-ray therapy; Particle-irradiation therapy A61N5/01 takes precedence
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: C12N 15/1024 In vivo mutagenesis using high mutation rate "mutator" host strains by inserting genetic material, e.g. encoding an error prone polymerase, disrupting a gene for mismatch repair
• CPC Class Codes: C12N 9/644 Coagulation factor IXa (3.4.21.22)
• CPC Class Codes: C12Y 304/21022 Coagulation factor IXa (3.4.21.22)

Abstract

Disclosed herein are methods and compositions for insertion of Factor IX (FIX) sequences into the genome of a cell for treating hemophilia B.

Claims

1. A method for treating hemophilia B in a subject, the method comprising integrating a sequence encoding a functional Factor IX (FIX) protein or fragment thereof into the genome of a cell using at least one zinc finger nuclease, wherein the zinc finger nuclease comprises an engineered zinc finger protein DNA-binding domain that binds to a target site in an endogenous Factor IX (FIX) gene, wherein the DNA-binding domain comprises four or five zinc finger recognition regions ordered F1 to F4 or F1 to F5 from N-terminus to C-terminus, and wherein

(i) when the DNA-binding domain comprises five zinc finger recognition regions, F1 to F5 comprise the following amino acid sequences:

2. The method of claim 1, wherein the sequence is integrated into an endogenous gene FIX gene.

3. The method of claim 1, wherein the sequence is delivered to the cell using a vector selected from the group consisting of a viral vector, a non-viral vector and combinations thereof.

4. The method of claim 1, wherein the at least one zinc finger nuclease is delivered to the cell using a vector selected from the group consisting of a viral vector, a non-viral vector and combinations thereof.

5. The method of claim 1, wherein the cell is a hepatic cell and the sequence is delivered to the cell by intravenous administration into the liver of an intact animal, intraperitoneal administration, direct injection into liver parenchyma, injection into the hepatic artery, or retrograde injection through the biliary tree.

6. The method of claim 1, further comprising the step of performing a partial hepatectomy on the subject.

7. The method of claim 1, further comprising the step of treating the subject with at least one secondary agent.

8. The method of claim 7, wherein the secondary agent is selected from the group consisting of gamma irradiation, UV irradiation, tritiated nucleotides, cis-platinum, etoposide, hydroxyurea, aphidicolin, prednisolone, carbon tetrachloride, adenovirus and combinations thereof.

9. The method of claim 1, wherein the cell is an isolated cell and the method further comprises administering the isolated cell to the subject.

10. The method of claim 9 wherein the zinc finger nuclease is delivered to the cell as an RNA.

11. The method of claim 1, wherein the subject is selected from the group consisting of an embryo, a fetus, a neonate, an infant, a juvenile or an adult.

12. The method of claim 1, further comprising associating the sequence with a homing agent that binds specifically to a surface receptor of the cell.

13. The method of claim 12, wherein the homing agent comprises galactose or a hybrid of an AAV coat protein and galactose.

14. The method of claim 1, further comprising associating a polynucleotide encoding the at least one nuclease with a homing agent that binds specifically to a surface receptor of the cell.

15. The method of claim 14, wherein the homing agent comprises galactose or a hybrid of an AAV coat protein and galactose.

16. The method of claim 1, wherein the cell is selected from the group consisting of a human cell, a nonhuman primate cell, a Rodenta cell, a Lagomorpha cell, a Carnivora cell and an Arteriodactyla cell.

17. The method of claim 1, wherein the cell is a stem cell.

18. The method of claim 17, wherein the stem cell is an embryonic stem cell, a hematopoietic stem cell, an induced pluripotent stem cell, a hepatocyte or a hepatic stem cell.