Patent application EP19173365A (EP), filed 2019-05-08
Publication number: EP3736330A1
Title: MODIFIED ADENO-ASSOCIATED VIRUS (AAV) PARTICLES FOR GENE THERAPY
Abstract: The present invention relates to improved adeno-associated virus (AAV) particles for gene delivery and gene therapy. Provided are adeno associated virus (AAV) particles that comprise a modified capsid. The present invention further relates to methods for producing the improved AAV particles of this invention by removing natural binding sites in adeno associated virus (AAV) capsids and introducing ligand binding sites into said capsid to provide AAVs that transduce only particular cells of interest. An additional aspect of the present invention relates to modified AAV particles for use in the treatment of a disease and methods for treating a disease, comprising administering the modified AAV particles to a subject in need thereof. Yet a further aspect of this invention relates to the AAV particles of this invention for the transfection of cells, for example as a gene delivery tool in basic research.
IPC codes: A61K  48/    00            A I
C12N  15/    86            A I
C12N   7/    00            A I
Claims: number: 1
text: Adeno associated virus (AAV) particle, comprising a modified capsid, wherein said modified capsid comprises at least one modification selected from the removal of a natural binding site in said capsid, and the introduction of a ligand binding site into said capsid.
number: 2
text: The AAV particle according to claim 1, wherein said AAV is selected from AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, and AAV12.
number: 3
text: The AAV particle according to claim 1 or 2, wherein at least one protein in said capsid is modified, preferably wherein said at least one protein is VP1, VP2, and/or VP3.
number: 4
text: The AAV particle according to any one of claims 1 to 3, wherein said modification of said capsid comprises the removal of at least one natural binding site and the introduction of at least one ligand binding site.
number: 5
text: The AAV particle according to any one of claims 1 to 4, wherein said natural binding site is a natural binding site that enables binding to heparan sulfate proteoglycans, and is preferably removed by replacing at least one of arginines 585 and 588 of VP1 and/or an analogous arginine in VP2 or VP3 with a different amino acid, such as alanine.
number: 6
text: The AAV particle according to any one of claims 1 to 5, wherein said ligand binding site is a ligand binding site that enables the covalent attachment of a ligand, and is preferably selected from a benzylguanine group, a benzylcytosine group, a chloroalkane group, an azide group, a phosphine, or combinations thereof.
number: 7
text: The AAV particle according to claim 6, further comprising a ligand that is attached to said benzylguanine group, said benzylcytosine group, said chloroalkane group, said azide group and/or said phosphine in particular a HaloTag™, a SNAP-tag™, or a CLIP-tag™, an azide or a phosphine.
number: 8
text: The AAV particle according to claim 6 or 7, wherein said ligand is selected from a protein ligand, a toxin subunit, a lectin, an adhesion factor, an antibody, a peptide, and a gene editing nuclease.
number: 9
text: A method of producing an improved adeno associated virus (AAV) particle, comprising the step of introducing at least one modification into a capsid of said AAV, preferably wherein said modification comprises introducing at least one ligand binding site into said capsid, optionally wherein a natural binding site in said capsid is removed.
number: 10
text: A pharmaceutical composition, comprising the AAV particle according to any one of claims 1 to 8, together with at least one pharmaceutically acceptable carrier and/or diluent.
number: 11
text: The AAV particle according to any one of claims 1 to 8 or the pharmaceutical composition according to claim 10 for use in the treatment of a disease, wherein said disease is preferably one that can be treated by gene therapy, such as cancer, an inherited monogenic disease, a genetic skin disease, an infectious disease, type I diabetes, and wound healing.
number: 12
text: A method for treating a disease that can be treated by gene therapy, comprising administering the AAV particle according to any one of claims 1 to 8 or the pharmaceutical composition according to claim 10 to a subject in need thereof, wherein said disease is preferably one that can be treated by gene therapy, such as cancer, an inherited monogenic disease, a genetic skin disease, an infectious disease, type I diabetes, and wound healing.
number: 13
text: Use of the AAV particle according to any one of claims 1 to 8 for the transfection of a cell, for example as a gene delivery tool.
number: 14
text: The use of the AAV particle according to claim 13, wherein said use is for cosmetic purposes.
Office action: search_report, dated 2020-11-11
Rejection reasons: novelty_epc_54
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severity: background
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severity: novelty_destroying
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severity: novelty_destroying
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severity: novelty_destroying
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severity: novelty_destroying
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ref_type: npl
severity: novelty_destroying
ref_id: - MATTHEW D STACHLER ET AL, "Site-specific Modification of AAV Vector Particles With Biophysical Probes and Targeting Ligands Using Biotin Ligase", MOLECULAR THERAPY, (20080617), vol. 16, no. 8, doi:10.1038/mt.2008.129, ISSN 1525-0016, pages 1467 - 1473, XP055187899 [X] 1-3,6,8-13 * see the whole document, in particular Figure 1 and section "Addition of targeting ligands to AAV vector particles with biotin ligase" *
ref_type: npl
severity: novelty_destroying
ref_id: - XIANGQUN SHI, "Insertional Mutagenesis at Positions 520 and 584 of Adeno-Associated Virus Type 2 (AAV2) Capsid Gene and Generation of AAV2 Vectors with Eliminated Heparin- Binding Ability and Introduced Novel Tropism", HUMAN GENE THERAPY, (20060301), vol. 17, pages 353 - 361, XP055507574 [X] 1-5,9-13 * see the whole document, in particular the cited passages; page  356, column  r, line  l  - page  357, column  l, line  2; figures 2,3; table 1 *
ref_type: npl
severity: novelty_destroying
Final disposition: pending (2020-11-11)