rAAV vector for the treatment of GM1 gangliosidosis and GM2 gangliosidosis
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- UNIV OF MASSACHUSETTS
- Filing Date
- 2026-01-07
- Publication Date
- 2026-06-09
Smart Images
Figure 2026094091000014 
Figure 2026094091000015 
Figure 2026094091000016
Abstract
Claims
1. Selected lysosome accumulation from HEXA and HEXB via chimeric introns. A promoter operably linked to a trans gene encoding a disorder-related protein. Capsid containing nucleic acids Recombinant AAV (rAAV) including this.
2. The recombinant AAV according to claim 1, wherein the nucleic acid does not contain an enhancer element.
3. The promoter comprises a chicken belta-actin (CB) promoter, according to the claim. rAAV as described in 1 or 2.
4. The chimeric intron is an intron sequence of chicken beta-actin, and / or The claim according to any one of claims 1 to 3, comprising the intron sequence of rabbit betaglobin rAAV.
5. The chimerintron is adjacent to an untranslated sequence, and optionally the untranslated sequence is a rabbit The rAA according to any one of claims 1 to 4, which is the untranslated sequence of gibert-globin. V.
6. The untranslated sequence of the rabbit betaglobin is derived from exon 1 or exon 2. The rAAV according to claim 5, which is an untranslated sequence.
7. The trans gene is distributed between the chimeric intron and the polyadenylation signal. The trans gene is placed, and optionally, the untranslated sequence and the polyadenylated signal rAAV according to any one of claims 1 to 6, which is positioned between the r and the r.
8. The HexA comprises the amino acid sequence specified in SEQ ID NO: 20, according to claims 1 to 7. rAAV as described in item 1.
9. The HexB comprises the amino acid sequence specified in SEQ ID NO: 21, according to claims 1 to 7. rAAV as described in item 1.
10. AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 , AAVrh8, AAV9, AAV10, and serotype selected from AAVrh10 rAAV according to any one of claims 1 to 9, comprising a capsid protein.
11. The following is a description of any one of claims 1 to 9, comprising the AAVrh8 serotype capsid protein rAAV.
12. It includes one or more ITRs, each ITR being AAV1 ITR, AAV2 ITR, From AAV3 ITR, AAV4 ITR, AAV5 ITR, and AAV6 ITR An rAAV according to any one of claims 1 to 11, selected from the group.
13. Isolated nucleic acids containing the sequences specified by SEQ ID NOs: 1-6 or 16-19.
14. Includes a sequence encoding a peptide having the sequence specified by SEQ ID NO: 20 or 21 isolated nucleic acid.
15. A host cell comprising the isolated nucleic acid construct according to claim 13 or 14.
16. The host cell according to claim 15, which is a eukaryotic cell.
17. The host cell according to claim 15 or 16, which is a mammalian cell.
18. The host cell according to claim 15, which is a prokaryotic cell.
19. The host cell according to claim 18, which is a bacterial cell.
20. Claims 15 to 19 further comprise isolated nucleic acids encoding AAV capsid proteins. The host cells described in any one of the items.
21. The capsid protein is either AAVrh8 capsid protein or AAVrh10 capsid protein. A host cell according to claim 20, which is a psidoprotein.
22. (i) via chimerintron, beta-hexosaminidase subunit alpha Includes a promoter operably linked to a trans gene encoding (HexA). The first nucleic acid is contained in a capsid, and optionally, the HexA is as shown in SEQ ID NO:
20. The first rAAV, including the amino acid sequence shown; (ii) via chimerintron, beta-hexosaminidase subunit beta Includes a promoter operably linked to a trans gene encoding (HexB). The product comprises a capsid containing a second nucleic acid, wherein HexB is optionally specified in SEQ ID NO:
21. The second rAAV, which includes the amino acid sequence shown, Includes, A pharmaceutical composition further comprising, optionally, a pharmaceutically acceptable carrier.
23. The first rAAV and / or the second rAAV have the AAVrh8 serotype. The pharmaceutical composition according to claim 22, comprising a capsid protein.
24. The first nucleic acid and / or the second nucleic acid surround the trans gene, AAV Claim 2, which includes an ITR, and optionally the AAV ITR is an AAV2 ITR. The pharmaceutical composition described in 2 or 23.
25. The first nucleic acid and / or the second nucleic acid does not contain an enhancer element. or the pharmaceutical composition according to any one of claims 22 to 24.
26. The promoter in (i) and / or the promoter in (ii) is a chicken The following is a description of any one of claims 22 to 25, comprising a ter-actin (CB) promoter. A pharmaceutical composition.
27. The chimera intron described in (i) and / or the chimera intron described in (ii) Intron sequences of rabbit betaactin, and / or intron sequences of rabbit betaglobin A pharmaceutical composition according to any one of claims 22 to 26, comprising a ron sequence.
28. The chimera intron described in (i) and / or the chimera intron described in (ii) is not The translation sequence is adjacent, and optionally the untranslated sequence is the untranslated sequence of rabbit betaglobin. A pharmaceutical composition according to any one of claims 22 to 27, wherein the composition is a row.
29. The untranslated sequence of the rabbit betaglobin is derived from exon 1 or exon 2. The pharmaceutical composition according to claim 28, wherein the sequence is untranslated.
30. The trans gene in (i) and / or the trans gene in (ii) It is positioned between the chimeric intron and the polyadenylation signal, and optionally the transistor The claim states that the lance gene is positioned between the untranslated sequence and the polyadenylation signal. A pharmaceutical composition according to any one of items 22 to 29.
31. The first rAAV and the second rAAV are present in the composition in a 1:1 ratio. The pharmaceutical composition according to any one of claims 22 to 30.
32. A method for treating lysosome storage disorders, any one of claims 1 to 12 Recombinant AAV (rAAV) as described in the item, or as described in any one of claims 22 to 31 A method comprising the step of administering a pharmaceutical composition to a subject having a lysosomal storage disorder.
33. Claim 3, wherein the lysosomal storage disorder is Tay-Sachs disease or Sandhoff disease. The method described in 2.
34. The rAAV or pharmaceutical composition is administered by intracranial injection, intracerebral injection, or to the ventricular system, cisterna magna, or The method according to claim 32 or 33, which is administered by injection into the CSF via the spinal canal. Law.
35. A container for storing recombinant AAV (rAAV) according to any one of claims 1 to 12 A kit that includes this.
36. (i) via chimerintron, beta-hexosaminidase subunit alpha Includes a promoter operably linked to a trans gene encoding (HexA). The first nucleic acid is contained in a capsid, and optionally, the HexA is as shown in SEQ ID NO:
20. A first container containing a first rAAV having the indicated amino acid sequence; (ii) via chimerintron, beta-hexosaminidase subunit beta Includes a promoter operably linked to a trans gene encoding (HexB). The product comprises a capsid containing a second nucleic acid, wherein HexB is optionally specified in SEQ ID NO:
21. A second container containing a second rAAV having the indicated amino acid sequence; optionally, (iii) A third container containing a pharmaceutically acceptable carrier and A kit that includes this.
37. The first container, the second container, and / or the third container are syringes, according to the claim. The kit described in 35 or 36.
38. Claim 32, wherein the administration comprises injection into the CSF via the cisterna magna and / or the cerebrospinal cavity. The method described in any one of items 34.
39. The method according to claim 38, wherein the administration further comprises an intrathalamic injection.
40. The aforementioned administration includes injection into the CSF via the cisterna magna and spinal cavity, and administration by intrathalamic injection. The method according to any one of claims 32 to 34 or 38 to 39.
41. The aforementioned administration is administered into the spinal canal via a fluoroscopically guided lumbar intrathecal catheter. The method according to any one of claims 32 to 34 or 38 to 40.
42. The total amount of rAAV administered to the subject is approximately 1 × 10⁻⁶ １１ ~1 x 10 １５ Vector genome (vg) During the copy, as described in any one of claims 32 to 34 or 38 to 41 Method of loading.
43. The subject, before administration of the rAAV or the pharmaceutical composition, in the CSF of the subject In this case, β-hexosaminidase activity less than 0.5% of normal β-hexosaminidase A activity. The method according to any one of claims 32 to 34 or 38 to 42, having A activity.
44. The subject exhibits normal yeast growth approximately three months after administration of the rAAV or the pharmaceutical composition. At least 0.5%, at least 1.0%, or at least 1.4% of the primary activity. Claims 32 to 3, which have an increased enzymatic activity of β-hexosaminidase A in CSF. The method described in any one of paragraphs 4 or 38 to 43.
45. The aforementioned subject is the enzyme activity of β-hexosaminidase A in CSF, before administration. An increase of at least two times, or at least three times, compared to the baseline of the subject. The method according to any one of claims 32 to 34 or 38 to 44.
46. Operablely linked to the trans gene encoding the human GLB1 sequence in SEQ ID NO:
23. Capsid containing nucleic acids including a promoter Recombinant AAV (rAAV) including this.
47. Operablely linked to the trans gene encoding the human GLB1 sequence in SEQ ID NO:
23. Promoter Isolated nucleic acids containing the above.
48. The promoter comprises a chicken belta-actin (CB) promoter, according to the claim. The rAAV described in 46, or the isolated nucleic acid described in claim 47.
49. AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 Selected from the group consisting of AAVrh8, AAV9, AAV10, and AAVrh10 The rAAV according to claim 46 or 48, comprising a serotype capsid protein.
50. The rAAV according to claim 49, comprising the AAV9 capsid protein.
51. It includes one or more ITRs, and each ITR is AAV1 ITR, AAV2 ITR, From AAV3 ITR, AAV4 ITR, AAV5 ITR, and AAV6 ITR rAAV according to any one of claims 46 or 48 to 50, selected from the group. 。
52. The rAAV according to claim 51, comprising one or more AAV2 ITRs.
53. The rAAV according to claim 46 or 48 to 52, and a pharmaceutically acceptable carrier A pharmaceutical composition containing the following:
54. A method for treating GM1 gangliosidosis, according to claim 46 or 48 From 52, the recombinant AAV (rAAV), or the pharmaceutical composition according to claim 53, A method comprising the step of administering to a subject having GM1 gangliosidosis.
55. The method according to claim 49, wherein the administration is intravenous administration.
56. The dosage is approximately 10 per kilogram (kg) of the subject's body weight. １１ ~10 １４ rAAV The method according to claim 54 or 55, wherein the vector genome (vg) is between copies.
57. A method for treating GM2 gangliosidosis, any one of claims 1 to 12 The rAAV described in the item, or the pharmaceutical composition described in any one of claims 22 to 31, A method comprising the step of administering to a subject having GM2 gangliosidosis.
58. GM2 ganglioside was found to be effective approximately three months after administration of the rAAV or the pharmaceutical composition. In the cerebrospinal fluid (CSF) of the subject, at least 15% from baseline, Reduced by at least 20%, or at least 25%, from claims 32 to 34, 38 The method described in either 45 or 57.
59. The dose administered to the thalamus is approximately 1 x 10⁻⁶ １１ ~1 x 10 １５ Claim 3, which is between vg The method described in any of 2 to 34, 38 to 45, or 57 to 58.
60. The dose administered into the cisterna magna and / or the cerebrospinal cavity is approximately 1 × 10⁻¹⁶ １１ ~1 x 10 １５ vg The one in between, according to any one of claims 32 to 34, 38 to 45, or 57 to 59. method.