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KR-20260066181-A - RAAV VECTORS FOR THE TREATMENT OF GM1 AND GM2 GANGLIOSIDOSIS

KR20260066181AKR 20260066181 AKR20260066181 AKR 20260066181AKR-20260066181-A

Abstract

Aspects of the present disclosure relate to compositions and methods for treating lysosomal storage diseases such as GM1 gangliosidosis, Tay-Sachs disease, and Sandhoff disease. In some embodiments, the composition comprises a viral vector encoding beta-galactosidase. In some embodiments, the composition comprises a viral vector encoding a beta-hexosaminidase subunit (e.g., HEXA, HEXB, or a combination thereof).

Inventors

  • 에스터베스, 미구엘, 세나

Assignees

  • 유니버시티 오브 매사추세츠

Dates

Publication Date
20260512
Application Date
20191004
Priority Date
20181005

Claims (17)

  1. Recombinant AAV (rAAV) comprising a capsid containing a nucleic acid containing the sequence presented in SEQ ID NO: 16.
  2. rAAV comprising a capsid protein of a serotype selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAV9, AAV10, and AAVrh10 in claim 1.
  3. In paragraph 2, rAAV containing AAV9 capsid protein.
  4. In paragraph 2, rAAV comprises one or more ITRs, wherein each ITR is selected from the group consisting of AAV1 ITR, AAV2 ITR, AAV3 ITR, AAV4 ITR, AAV5 ITR, and AAV6 ITR.
  5. In paragraph 4, rAAV comprising one or more AAV2 ITRs.
  6. Isolated nucleic acid containing the sequence presented in SEQ ID NO: 16.
  7. Isolated host cell containing the isolated nucleic acid of paragraph 6.
  8. In paragraph 7, an isolated host cell that is a eukaryotic cell.
  9. In paragraph 7, the isolated host cell is a mammalian cell.
  10. In paragraph 7, an isolated host cell that is a prokaryotic cell.
  11. In paragraph 10, an isolated host cell that is a bacterial cell.
  12. A separated host cell comprising a separated nucleic acid encoding an AAV capsid protein, in any one of claims 7 to 9.
  13. In paragraph 12, isolated host cells in which the AAV capsid protein is the AAV9 capsid protein.
  14. A pharmaceutical composition comprising rAAV of any one of claims 1 to 5 and a pharmaceutically acceptable carrier.
  15. In claim 14, a pharmaceutical composition for use in treating GM1 gangliosidosis.
  16. In item 15, a pharmaceutical composition administered intravenously.
  17. A pharmaceutical composition according to claim 16, wherein the dose is about 10¹¹ to 10¹⁴ rAAV vector genome (vg) copies per kg of body weight of the subject.

Description

RAAV Vectors for the Treatment of GM1 and GM2 Gangliosidosis Related applications This application is based on 35 U.S.C. U.S. provisional patent application serial number 62/741,848 filed on October 5, 2018, under § 119(e) titled "RAAV vector for the treatment of GM1 and GM2 gangliosidosis", 62/814,587 filed on March 6, 2019, titled "RAAV vector for the treatment of GM1 and GM2 gangliosidosis", 62/815,996 filed on March 8, 2019, titled "RAAV vector for the treatment of GM1 and GM2 gangliosidosis", 62/826,863 filed on March 29, 2019, titled "RAAV vector for the treatment of GM1 and GM2 gangliosidosis", April 2019, titled "RAAV vector for the treatment of GM1 and GM2 gangliosidosis" Claiming the benefit of the filing dates of 62/840,359 filed on the 29th and 62/848,858 filed on May 16, 2019 under the title "RAAV vector for the treatment of GM1 and GM2 gangliosidosis," the full contents of each application are incorporated herein by reference. Reference to the list of sequences electronically submitted via EFS-WEB The present application comprises a sequence listing submitted electronically via EFS-Web (title: "Sequence Listing"; size: 80,237 bytes; and written in Patent-In 3.5), the entirety of which is incorporated herein by reference. field The present disclosure provides a recombinant adeno-associated virus (rAAV), a composition, and a kit useful for the treatment of lysosomal storage diseases in some embodiments. In some embodiments, the lysosomal storage disease is GM1 or GM2 gangliosidosis (e.g., Tay-Sachs disease). Federally funded research This invention was made with government support under Award No. HD060576, awarded by the National Institutes of Health. The U.S. government holds specific rights to this invention. GM1 gangliosidosis is an autosomal recessive neuropathic lysosomal storage disease caused by mutations in the human gene GLB1 . The human GLB1 gene encodes β-D-galactosidase (β-gal), an enzyme that removes terminal galactose residues from numerous molecules in the central nervous system (e.g., GM1 gangliosides) and peripheral tissues (e.g., oligosaccharides, glycoproteins, and glycosaminoglycans). A deficiency in lysosomal β-gal activity causes GM1 gangliosides and their asiallo-derived GA1 to accumulate primarily in the central nervous system, where their synthesis rate is highest. GM1 gangliosidosis is consistently fatal, and there is no effective treatment. Tay-Sachs disease (GM2 gangliosidosis) and Sandhoff disease are autosomal recessive lysosomal storage diseases caused by mutations in the HEXA and HEXB genes, respectively, resulting in a deficiency of β-N-acetyl-D-hexosaminidase (Hex) enzyme activity. The Hex enzyme is a heterodimer containing HexA and HexB protein subunits. The deficiency of Hex activity leads to the progressive accumulation of GM2 gangliosides in the central nervous system, resulting in neurodegeneration. Tay-Sachs disease is characterized by the continuous decline of the nervous system and eventual death. Symptoms of Tay-Sachs disease include frequent seizures, dysphagia, loss of motor control, and frequent respiratory infections. Sandhoff disease is also characterized by the progressive decline of the CNS and results in gait abnormalities, swallowing and speech disorders, peripheral neuropathy, psychiatric symptoms, and eventual death. Currently, there are no disease-transforming treatment options for Tay-Sachs disease or Sandhoff disease, and only symptomatic treatment including anticonvulsants and antispasmodics is available. Figures 1A and 1B show the design of the rAAV vector CBA-mβgal-WPRE. Figure 1A shows a schematic diagram of the rAAV vector CBA-mβgal-WPRE. Two reverse terminal repeats (ITRs) from AAV2 are located on the sides of the vector at each end. The CBA promoter consists of a cytomegalovirus immediate early enhancer (CMV) fused to a chicken beta-actin promoter, followed by a chimeric chicken beta-actin/rabbit beta-globin intron (CBA), mouse lysosomal acid β-galactosidase cDNA (mβgal), Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), and two poly-A signals in tandem derived from bovine growth hormone (BGH) and SV40. This vector was subsequently packaged into an AAVrh8 capsid. Figure 1B shows a recombinant AAV (rAAV) vector containing a transgene encoding hGLB1 under the control of a CBA promoter. Figures 2A–2J show that intracranial injection of AAVrh8-mβgal into βgal -/- mice produces a dose-dependent enzyme distribution. βgal expression in the brains of representative AAVrh8-injected animals and age-matched controls was analyzed by histochemical staining with X-gal and counterstaining with Nuclear Fast Red on 20 μm coronal brain sections at 2 weeks (4e10vg) or 3 months (2.6e10vg and 2.6e9vg) after injection. Figures 2A–2B represent 4e10vg; Figures 2E–2F represent 2.6e10vg; Figures 2I–2J represent 2.6e9vg; Figures 2C–2D represent naive βgal -/- ; and Figures 2G–2H represent naive βgal +/- mice. Images represent N ≥ 3 mic