AAV vector variants for ocular gene delivery

Patent metadata

Field	Value
Publication number	US-12509705-B2
Application number	US-202017755434-A
Country	US
Kind code	B2
Filing date	Nov 2, 2020
Priority date	Oct 31, 2019
Publication date	Dec 30, 2025
Grant date	Dec 30, 2025

Abstract

The present invention relates to adeno-associated virus capsid polypeptide sequences and their use in therapeutic transgene delivery to the eye and potentially other tissues.

First claim

The invention claimed is:

1 . An adeno-associated virus (AAV) capsid polypeptide comprising a peptide insert in the C-terminal direction at position 453 or in the C-terminal direction at position 587, 588, 589, 590, 591 or 592 of the AAV serotype 2 capsid having amino acid sequence of SEQ ID NO: 1 or a position homologous thereto in an AAV of another serotype, wherein the peptide insert is selected from:

(Cap3; SEQ ID NO: 10)

SASEAST,  

(Cap5; SEQ ID NO: 11)

DTRPHDQ,

(Cap7; SEQ ID NO: 12)

EHYNSTC, 

(Cap9; SEQ ID NO: 13)

PNPNCTL, 

(Cap11; SEQ ID NO: 14)

TPPSITA,

(Cap12; SEQ ID NO: 15)

CGESSYL, 

(Cap13; SEQ ID NO: 16)

PRTPHTA 

and 

(Cap14; SEQ ID NO: 17)

ELCDGFA.

2 . The AAV capsid polypeptide of claim 1 , wherein the insert is at position 587, 588, 589, 590, 591 or 592 of the AAV serotype 2 capsid or a position homologous thereto in an AAV of another serotype.

3 . The AAV capsid polypeptide of claim 2 , wherein the insert is at position 587 of the AAV serotype 2 capsid or a position homologous thereto in an AAV of another serotype.

4 . The AAV capsid polypeptide of claim 1 , wherein the insert is selected from

(Cap3; SEQ ID NO: 10)

SASEAST, 

(Cap11; SEQ ID NO: 14)

TPPSITA,

(Cap13; SEQ ID NO: 16) 

PRTPHTA and 

(Cap14; SEQ ID NO: 17)

ELCDGFA.

5 . The AAV capsid polypeptide of claim 1 , wherein the polypeptide comprises an insert in the C-terminal direction at position 587.

6 . The adeno-associated virus capsid polypeptide of claim 1 , wherein the AAV capsid protein is an AAV2 capsid comprising:
a. one or more tyrosine to phenylalanine substitutions at positions 252, 272, 444, 500, 700, 704 and 730, and/or b. one or more threonine to valine substitutions.

7 . The AAV capsid polypeptide of claim 6 , wherein the one or more tyrosine to phenylalanine substitutions are at positions 252, 272, 444, 500, 704 and 730.

8 . The AAV capsid polypeptide of claim 6 , wherein the one or more tyrosine to phenylalanine substitutions are at all of the positions 252, 272, 444, 500, 700 and 730.

9 . The AAV capsid polypeptide of claim 6 , wherein the one or more threonine to valine substitutions is T491V.

10 . A nucleic acid sequence encoding the AAV capsid polypeptide according to claim 1 .

11 . The nucleic acid sequence of claim 10 , wherein the sequence is a self-complementary or single stranded vector genome.

12 . The nucleic acid sequence of claim 11 , wherein the sequence is a self-complementary vector genome.

13 . The nucleic acid sequence of claim 10 or claim 11 , wherein the nucleic acid sequence comprises a transgene, wherein the transgene encodes a light-sensitive protein, a siRNA, a shRNA or a CRISPR/Cas-gRNA cassette.

14 . The nucleic acid sequence of claim 13 , wherein the transgene is under control of a promoter sequence operable in a mammalian cell.

15 . The nucleic acid sequence of claim 14 , wherein the mammalian cell is a retinal cell.

16 . The nucleic acid sequence of claim 15 , wherein the retinal cell is a human retinal cell.

17 . The nucleic acid sequence of claim 14 , wherein the promoter is a ubiquitous or cell-specific promoter.

18 . The nucleic acid sequence of claim 14 , wherein the promoter is selected from a CMV immediate early promoter, or hEfla promoter.

19 . A method of treating a condition in a subject in need thereof, comprising administering the AAV capsid polypeptide of claim 1 , wherein the condition affects a retinal or retinal pigment epithelium cell, a photoreceptor, a bipolar cell, a ganglion cell, and/or an amacrine cell.

20 . The method of claim 19 , wherein the AAV capsid polypeptide is administered by intravitreal administration or by subretinal administration.

21 . The method of claim 19 , wherein the intravitreal administration is intravitreal injection.

Assignees

• Univ Bern
• Medizinische Hochschule Hannover

Inventors

• Kleinlogel Sonja
• David Anand
• Büning Hildegard

Classifications

• C12N2810/40

  Vectors comprising a peptide as targeting moiety, e.g. a synthetic peptide, from undefined source · CPC title

• C12N2750/14145

  Special targeting system for viral vectors · CPC title

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title

• C12N2750/14122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• C07K2319/33

  fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies · CPC title