Search

CA-3124880-C - GENE THERAPY CONSTRUCTS FOR TREATING WILSON DISEASE

CA3124880CCA 3124880 CCA3124880 CCA 3124880CCA-3124880-C

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.

Inventors

  • Christine Livingston
  • Samuel Wadsworth

Assignees

  • ULTRAGENYX PHARMACEUTICAL INC.

Dates

Publication Date
20260505
Application Date
20200103
Priority Date
20190104

Claims (20)

  1. WHAT IS CLAIMED IS: 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 a truncated human copper-transporting ATPase 2 (ATP7B) having the amino acid sequence set forth in SEQ ID NO:8; and (d) a 3’-ITR sequence.
  2. 2. The recombinant nucleic acid construct according to claim 1, 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 promoter sequence, a thyroxine binding globulin (TBG) promoter sequence, an alpha-1 anti-trypsin (A1AT) promoter sequence, and a CAG promoter sequence.
  3. 3. The recombinant nucleic acid construct according to claim 2, wherein the promoter sequence is the TTR promoter sequence.
  4. 4. The recombinant nucleic acid construct according to any one of claims 1-3, wherein the 5’-ITR sequence is from AAV2.
  5. 5. The recombinant nucleic acid construct according to any one of claims 1-3, wherein the 3’-ITR sequence is from AAV2.
  6. 6. The recombinant nucleic acid construct according to any one of claims 1-3, wherein the 5’-ITR sequence and the 3’-ITR sequence are from AAV2.
  7. 7. The recombinant nucleic acid construct according to any one of claims 1-6, wherein the 5’-ITR sequence and the 3’-ITR sequence comprises or consists of SEQ ID NO:2. 33
  8. 8. The recombinant nucleic acid construct according to any one of claims 1-3, wherein the 5’-ITR sequence and/or the 3’-ITR sequence are from a non-AAV2 source.
  9. 9. The recombinant nucleic acid construct according to any one of claims 1-8, wherein the recombinant nucleic acid construct further comprises one or more enhancer sequences.
  10. 10. The recombinant nucleic acid construct according to claim 9, wherein the 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.
  11. 11. The recombinant nucleic acid construct according to claim 10, wherein the enhancer sequence is the enTTR enhancer sequence.
  12. 12. The recombinant nucleic acid construct according to claim 11, wherein the enhancer sequence comprises or consists of SEQ ID NO:3.
  13. 13. The recombinant nucleic acid construct according to any one of claims 10-12, wherein the enhancer sequence is located upstream of the promoter sequence.
  14. 14. The recombinant nucleic acid construct according to any one of claims 1-13, wherein the recombinant nucleic acid construct further comprises one or more intron sequences.
  15. 15. The recombinant nucleic acid construct according to claim 14, wherein the intron sequence 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/IgG chimeric intron sequence, and an hFIX intron sequence.
  16. 16. The recombinant nucleic acid construct according to claim 15, wherein the intron sequence is the SV40 Small T intron sequence. 34
  17. 17. The recombinant nucleic acid construct according to claim 16, wherein the intron sequence comprises or consists of SEQ ID NO:4.
  18. 18. The recombinant nucleic acid construct according to claim 15, wherein the intron sequence is the rHBB intron sequence.
  19. 19. The recombinant nucleic acid construct according to claim 18, wherein the intron sequence comprises or consists of SEQ ID NO:5.
  20. 20. The recombinant nucleic acid construct according to any one of claims 1-19, wherein the recombinant nucleic acid construct further comprises a polyadenylation signal sequence.

Description

GENE THERAPY CONSTRUCTS FOR TREATING WILSON DISEASE CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of and priority to U.S. Provisional Patent Application No. 62/788,324, filed January 4, 2019; and to U.S. Provisional Patent Application 5 No. 62/834,830, filed April 16, 2019. SEQUENCE LISTING [0002] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format. Said ASCII copy, created on December 31, 2019, is named ULP-003WO_SL_ST25.txt and is 49,846 bytes in size. TECHNICAL FIELD OF THE INVENTION [0003] This application relates generally to adeno-associated viral vectors and methods of their use in gene therapy for treating Wilson disease (WD ). BACKGROUND OF THE INVENTION [0004] Wilson disease (WD) is an autosomal recessive genetic disorder that causes 15 accumulation of copper primarily in the liver and subsequently in the neurological system and other tissues. WD is a rare disorder that affects approximately 1 in 30,000 individuals, caused by mutations in the copper transporting ATPase 2 (ATP7B) gene on chromosome 13. There are more than 600 unique ATP7B mutations. ATP7B is expressed mainly in hepatocytes and functions in the transmembrane transport of copper. Absent or reduced 20 function of ATP7B protein results in decreased hepatocellular excretion of copper into bile, causing liver disease. Over time without proper treatment, high copper levels can cause lifethreatening organ damage. [0005] Patients with hepatic WD usually present in late childhood or adolescence, and exhibit features of acute hepatitis, fulminant hepatic failure, or progressive chronic liver 25 disease. Neurologic manifestations ofWD typically present later than the liver disease, most often in the second or third decade and include extrapyrarnidal, cerebellar, and cerebralrelated symptoms. 1 Date Re9ue/Date Received 2024-04-03 WO 2020/142653 PCT/0S2020/012131 [0006] The aim of medical treatment of WD is to remove the toxic deposit of copper from the body and to prevent its reaecumulation. Current treatment approaehes for WD are daily oral therapy with chelating agents (D-penicillamine, trientine, and zinc salts). Medical therapy is effective in most, but not all WD patients. Liver transplantation is a therapeutic 5 option in WD patients presenting with fulminant liver failure or progressive liver failure. However, transplant recipients are required to maintain a constant immune suppression regimen to prevent rejection. [0007] The present invention addresses the need for improved and sustainable treatment of WD by delivering a gene expressing truncated yet functional A TP7B to patients with an 10 adeno-associated viral vector. The truncated A TP7B of the present invention has improved efficacy in treating WD and possesses an advantage of manufacturing ease and efficiency over wild-type and other truncated forms of A TP7B protein. SUMMARY OF THE INVENTION [0008] This invention provides compositions and methods of their use in gene therapy. 15 Provided herein arc adcno-associatcd virus (AA V) vectors useful for the treatment of WD. In one aspect, the present invention provides a recombinant nucleic aeid construct comprising: a 5'-invcrtcd terminal repeat (ITR) sequence; a promoter sequence; a nucleic acid sequence encoding a truncated human copper-transporting ATPasc 2 (A TP7B) in which metal-binding domains (MBDs) 1-3 have been deleted, but the serine-rich loop including two serine 20 residues (S340 and S341) between MBD3 and MBD4 is present; and a 3'-ITR sequence. [0009] In another aspect, the present invention provides a recombinant adeno-associatcd virus (rAA V) useful for the treatment of Wilson disease, in which rAA V comprises an AA V capsid and a vector genome packaged therein, the vector genome comprises a 5 '-inverted terminal repeat (ITR) sequence; a promoter sequence; a nucleic acid sequence encoding a 25 truncated human copper-transporting ATPase 2 (ATP7B) 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 a 3'-ITR sequence. [0010] These and other aspects and features of the invention arc described in the following sections of the application. 2 WO 2020/142653 PCT/0S2020/012131 [0011] drawings. BRIEF DESCRIPTION OF lHE DRAWINGS The invention can be more completely understood with reference to the following FIG. 1 is an illustrative diagram showing an exemplary vector genome construct 5 comprising a nucleotide sequence, which encodes a truncated human copper-transporting ATPase 2 (ATP7B), in which the metal-binding domains (MBDs) 1-3 have been deleted, but the serine-rich loop including two serine residues (S340 and S34 l) between MBD3 and MBD4 is present ("ATP7B .1.1-3-SS" or "ATP7B dell-3 native"). Features of the exemplary vector genome construct are provided below: Start (Nucleotide Position) End (Nucleotide Position) 1