Methods and compositions for dual glycan binding aav vectors

Patent metadata

Field	Value
Publication number	US-2016017005-A1
Application number	US-201414777070-A
Country	US
Kind code	A1
Filing date	Mar 14, 2014
Priority date	Mar 15, 2013
Publication date	Jan 21, 2016
Grant date	—

Abstract

The present invention provides methods and compositions comprising an adeno-associated virus (AAV) capsid protein, comprising one or more amino acids substitutions, wherein the substitutions introduce a new glycan binding site into the AAV capsid protein.

First claim

1 . An adeno-associated virus (AAV) capsid protein, comprising one or more amino acids substitutions, wherein the substitutions introduce a new glycan binding site into the AAV capsid protein.

2 . The AAV capsid protein of claim 1 , wherein the amino acid substitutions are in amino acid 266, amino acids 463-475 and amino acids 499-502 in AAV2 or the corresponding amino acid positions in AAV1, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 or AAV10.

3 . The AAV capsid protein of claim 1 , wherein the new glycan binding site is a hexose binding site, wherein the hexose is a galactose (Gal), a mannose (Man), a glucose (Glu) or a fucose (fuc).

4 . The AAV capsid protein of claim 1 , wherein the new glycan binding site is a sialic acid (Sia) binding site, wherein the Sia residue is N-acetylneuraminic acid (Neu5Ac) or N-Glycolylneuraminic acid (Neu5Gc).

5 . The AAV capsid protein of claim 1 , wherein the new glycan binding site is a disaccharide binding site, wherein the disaccharide is a sialic acid linked to galactose in the form Sia(alpha2,3)Gal or Sia(alpha2,6)Gal.

6 . The AAV capsid protein of claim 3 , wherein the new glycan binding site is a galactose binding site.

7 . The AAV capsid protein of claim 1 , wherein the substitutions introduce a new glycan binding site from a first AAV serotype into the capsid protein of a second AAV serotype that is different from said first AAV serotype.

8 . The AAV capsid protein of claim 7 , wherein the serotype of the second AAV serotype is AAV serotype 1 (AAV1), AAV serotype 2 9AAV2), AAV serotype 3a (AAV3a), AAV serotype 3b (AAV3b), AAV serotype 4 (AAV4), AAV serotype 5 (AAV5), AAV serotype 6 (AAV6), AAV serotype 7 (AAV7), AAV serotype 8 (AAV8), or AAV serotype 10 (AAV10).

9 . The AAV capsid protein of claim 7 , wherein the new glycan binding site is the galactose binding site from AAV serotype 9 (AAV9).

10 . The AAV capsid protein of claim 2 , wherein the AAV capsid protein is from AAV2, and
a) the substitution at amino acid 266 is A266S; b) the substitutions at amino acids 463-475 are SQAGASDIRDQSR463-475SX 1 AGX 2 SX 3 X 4 X 5 X 6 QX 7 R, wherein X 1-7 can be any amino acid; and c) the substitutions at amino acids 499-502 are EYSW499-502EX 8 X 9 W, wherein X 8-9 can be any amino acid.

11 . The AAV capsid protein of claim 10 , wherein: X 1 is V; X 2 is P; X 3-6 are NMAV; and X 7 is G.

12 . The AAV capsid protein of claim 10 , X 8 is F and X 9 is W.

13 . An AAV capsid comprising the AAV capsid protein of claim 1 .

14 . A virus vector comprising:
(a) the AAV capsid of claim 13 ; and (b) a nucleic acid comprising at least one terminal repeat sequence, wherein the nucleic acid is encapsidated by the AAV capsid.

15 . A composition comprising the virus vector of claim 14 in a pharmaceutically acceptable carrier.

16 . A method of introducing a nucleic acid into a cell, comprising contacting the cell with the virus vector of claim 14 .

17 . The method of claim 16 , wherein the cell is in a subject.

18 . The method of claim 17 , wherein the subject is a human subject.

Assignees

• Univ North Carolina

Inventors

• Asokan Aravind
• Samulski Richard

Classifications

• A61P31/20

  for DNA viruses · CPC title

• A61P31/18

  for HIV · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P31/12

  Antivirals · CPC title

• A61P31/22

  for herpes viruses · CPC title