RAAV with chemically modified capsid
The invention is directed to a recombinant Adeno-Associated Virus (rAAV) vector particle having at least one primary amino group contained in the capsid proteins, chemically coupled with at least one ligand L, wherein coupling of said ligand L is implemented through a bond comprising a —CSNH— bond and an optionally substituted aromatic moiety. The present invention further relates to a method for chemically coupling an Adeno-Associated Virus (AAV) vector particle with at least one ligand L and to a Recombinant Adeno-Associated Virus (rAAV) vector particle obtained by said method as well as a pharmaceutical composition comprising it and their corresponding medical use.
1. A recombinant Adeno-Associated Virus (rAAV) vector particle having at least one primary amino group contained in the capsid proteins, chemically coupled with at least one ligand L, wherein said ligand L is implemented under the form of a compound of formula (I)
with
N* being the nitrogen atom of one primary amino group contained in the capsid proteins, and
representing an optionally substituted arylene or a heteroarylene radical, directly or not, covalently bound to at least one ligand L.
2. The recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 , wherein
represents a phenylene group, a napthylene or a pyridylene group.
3. The recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 , wherein said Adeno Associated Virus vector is composed of wild-type capsid proteins from naturally occurring serotypes.
4. The recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 , wherein said Adeno Associated Virus is an Adeno Associated Virus with a genetic modification of the capsid proteins from naturally occurring serotypes or composed by a synthetic capsid.
5. The recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 , wherein ligand L is a targeting agent, a steric shielding agent for avoiding neutralizing antibody interactions, a labeling agent or a magnetic agent.
6. The recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 , wherein ligand L is a targeting ligand derived from proteins, from mono- or polysaccharides, from steroid hormones, from RGD motif peptide, from vitamins, from small molecules or from targeting peptide.
7. The recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 , wherein ligand L is a steric shielding agent for avoiding neutralizing antibody interactions derived from synthetic polymers.
8. The recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 , wherein ligand L is a labeling agent for analytic gene transfer with fluorescent dye or nano gold particles, or radioactive dye.
9. The recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 , wherein ligand L is a magnetic agent.
10. The recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 , wherein the moiety of formula (I) is selected from the group consisting of
11. A method for chemically coupling an Adeno-Associated Virus (AAV) vector particle with at least one ligand L, said method comprising at least the steps of:
having an Adeno-Associated Virus (AAV) particle having at least one primary amino group contained in the capsid proteins, and
contacting said AAV particle with a reagent of formula (II)
B—N═C═S (II)
wherein B is a radical (L)m(X)Ar—
with
L being as defined in claim 5 ,
m representing an integer from 1 to 1000,
Ar representing an arylene or heteroarylene radical, and
X representing a bond or a spacer between said ligand(s) L and Ar, in conditions suitable for reacting the primary amino group with the —N═C═S moiety of the reagent of formula (II).
12. A method according to claim 11 , wherein X represents:
a group
wherein n represents a integer from 0 to 5000, or
a group
wherein n represents an integer from 0 to 5000.
13. The method according to claim 11 , wherein the step of contacting said AAV particle with a reagent of formula (II) takes place in presence of a buffer selected from Tris Buffered Saline, sodium carbonate-sodium bicarbonate buffer, PBS, and dPBS.
14. The method according to claim 11 , wherein the reaction takes place at a pH comprised between 7 and 9.6.
15. The method according to claim 11 , wherein the obtained Recombinant Adeno-Associated Virus (rAAV) vector particle is further reacted by modifying the capsid proteins in a second coupling step, by chemical coupling with unreacted amino groups from the first coupling step.
16. The method according to claim 11 , wherein said method comprises obtaining an Adeno-Associated Virus particle with altered tropism for accentuating the transduction of a specific cell or tissue or for decreasing the interaction with neutralizing antibodies.
17. A recombinant Adeno-Associated Virus (rAAV) vector particle obtained by a method according to claim 11 .
18. A method for delivering the recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 to a subject in need thereof, the method comprising administering to the subject a composition comprising said rAAV vector particle.
19. A method for targeting a desired specific organ, tissue or cell types in a subject in need thereof with the recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 , the method comprising at least a step of administering to said subject a composition comprising said rAAV vector particle, wherein the Adeno Associated Virus (AAV) has altered tropism for targeting to a desired specific organ, tissue or cell type.
20. A method for decreasing interaction with neutralizing antibodies with the recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 in a subject in need thereof, the method comprising at least a step of administering to said subject a composition comprising said rAAV vector particle, wherein said rAAV vector particle has a decreased interaction with neutralizing antibodies.
21. A method for targeting hepatocytes, retina, lung, heart, kidney, liver, brain, spleen, tumor or muscle cells, Retinal Pigmented Epithelium, photoreceptors, myocytes or cardiomyocytes with the recombinant Adeno-Associated Virus (rAAV) vector particle according to claim 1 in a subject in need thereof, the method comprising at least a step of administering to said subject a composition comprising said rAAV vector particle, wherein said Recombinant Adeno-Associated Virus (rAAV) vector particle has a selective tropism for hepatocytes, retina, lung, heart, kidney, liver, brain, spleen, tumor or muscle cells.
22. A pharmaceutical composition comprising the recombinant Adeno-Associated Virus (rAAV) vector particle as recited in claim 1 in a pharmaceutically acceptable carrier.