IP Library Granted Patent US 12,338,450
Granted Patent B2
US 12,338,450 · App. 17/417,619 · Granted Jun 24, 2025

Gene therapy constructs for treating Wilson disease

Inventors: Christine Livingston (Lambertville, NJ); Samuel Wadsworth (Shrewsbury, MA)
Assignee: Ultragenyx Pharmaceutical Inc.
C12N15/86A61K48/0058A61P43/00
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Quick Facts
Patent No.
US 12,338,450
App. No.
17/417,619
Granted
Jun 24, 2025
Kind
B2
Abstract

This application relates to adeno-associated viral vectors encoding a truncated yet functional ATP7B for use in gene therapy for treating Wilson disease (WD). The truncated ATP7B described herein has several advantages over the wild-type ATP7B such as higher efficacy and improved manufacturing yield.

Claims (33)

1. A recombinant nucleic acid construct comprising:

(a) a 5′-inverted terminal repeat (ITR) sequence;

(b) a promoter sequence;

(c) a nucleic acid sequence encoding the truncated human copper-transporting ATPase 2 (ATP7B) of SEQ ID NO: 8, in which metal-binding domains (MBDs) 1-3 have been deleted, but the serine-rich loop including two serine residues (S340 and S341) between MBD3 and MBD4 is present; and

(d) a 3′-ITR sequence.

2. A recombinant adeno-associated virus (rAAV) useful for the treatment of Wilson disease, said rAAV comprising an AAV capsid and a vector genome packaged therein, said vector genome comprising:

a. an AAV 5′-inverted terminal repeat (ITR) sequence;

b. a promoter sequence;

c. a nucleic acid sequence encoding the truncated human copper-transporting ATPase 2 (ATP7B) of SEQ ID NO: 8, in which metal-binding domains (MBDs) 1-3 have been deleted, but the serine-rich loop including two serine residues (S340 and S341) between MBD3 and MBD4 is present; and

d. an AAV 3′-ITR sequence.

3. The rAAV of claim 2 , wherein the AAV capsid is from an AAV of serotype 9, 8, 1, 2, 3, 4, 5, 6, 7, 10, 11, 12, rh10, or hu37.

4. The rAAV of claim 2 , wherein the promoter sequence is selected from a transthyretin (TTR) promoter sequence, a chicken β-actin (CBA) promoter sequence, a cytomegalovirus (CMV) immediate early gene-(CMV) promoter sequence, a thyroxine binding globulin (TBG) promoter sequence, an alpha-1 anti-trypsin (A1AT) promoter sequence, and a CAG promoter sequence.

5. The rAAV of claim 2 , wherein the AAV 5′-ITR sequence and/or the AAV 3′-ITR sequence is from AAV2.

6. The rAAV of claim 2 , wherein the AAV 5′-ITR sequence and/or the AAV 3′-ITR sequence are from a non-AAV2 source.

7. The rAAV of claim 2 , wherein the vector genome further comprises an enhancer sequence, and wherein said enhancer sequence is selected from a transthyretin enhancer (enTTR) sequence, a cytomegalovirus (CMV) immediate early gene enhancer sequence, a chicken β-actin (CBA) enhancer sequence, an En34 enhancer sequence, and an apolipoprotein (ApoE) enhancer sequence.

8. The rAAV of claim 7 , wherein the enhancer sequence is located upstream of the promoter sequence.

9. The rAAV of claim 2 , wherein the vector genome further comprises one or more intron sequences.

10. The rAAV of claim 9 , wherein the one or more intron sequences is selected from an SV40 Small T intron sequence, a rabbit hemoglobin subunit beta (rHBB) intron sequence, a human beta globin IVS2 intron sequence, a β-globin/IG chimeric intron sequence, and an hFIX intron sequence.

11. The rAAV of claim 2 , wherein the vector genome further comprises a polyadenylation signal sequence.

12. The rAAV of claim 11 , wherein the polyadenylation signal sequence is selected from an SV40 polyadenylation signal sequence, a bovine growth hormone (BGH) polyadenylation signal sequence, and a rabbit beta globin polyadenylation signal sequence.

13. A recombinant adeno-associated virus (rAAV) comprising an AAV capsid and a vector genome packaged therein, said vector genome comprising:

a. the AAV 5′-inverted terminal repeat (ITR) sequence of SEQ ID NO:2;

b. the enhancer sequence of SEQ ID NO:3;

c. the promoter sequence of SEQ ID NO: 12;

d. the nucleic acid sequence of SEQ ID NO:1 or SEQ ID NO: 15 encoding a truncated human copper-transporting ATPase 2 (ATP7B); and

e. the AAV 3′-ITR sequence of SEQ ID NO:2.

14. A composition comprising the rAAV of claim 13 , and a pharmaceutically acceptable carrier.

15. A method of treating Wilson disease in a human subject comprising intravenously administering to the human subject a dose of about 1×10 11 genome copies (GC)/kg to about 1×10 14 genome copies of the rAAV of claim 2 ,

wherein the AAV capsid is selected from: serotype 8 and serotype 9, and

wherein the promoter sequence is selected from the group consisting of: a transthyretin TTR) promoter sequence, a chicken β-actin (CBA) promoter sequence, a cytomegalovirus (CMV) immediate early gene promoter sequence, a thyroxine binding globulin (TBG) promoter sequence, an alpha-1 anti-trypsin (A1AT) promoter sequence, and a CAG promoter sequence.

16. The method of claim 15 , wherein the rAAV is administered at a dose of about 1×10 12 GC/kg to about 1×10 13 GC/kg.

17. The method of claim 15 , wherein administering the rAAV comprises administration of a single dose of rAAV.

18. The method of claim 15 , wherein administering the rAAV comprises administration of multiple doses of rAAV.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 28, 2022
From: LIVINGSTON, CHRISTINE; WADSWORTH, SAMUEL
To: ULTRAGENYX PHARMACEUTICAL INC.
Reel/Frame 059119/0430 →
Continuity (3)
Provisional Application 62834830 · Apr 16, 2019
Provisional Application 62788324 · Jan 4, 2019
Related Publication 20220090131A1 · Mar 24, 2022
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