AAV/UPR-plus virus, UPR-plus fusion protein, genetic treatment method and its use in treatment of neurodegenerative diseases, such as Parkinson's disease and Huntington's disease, among others

• US 10,889,831 B2
• Filed: 11/04/2016
• Issued: 01/12/2021
• Est. Priority Date: 11/04/2015
• Status: Active Grant

• Current Assignee: Universidad de Chile
• Original Assignee: Universidad de Chile
• Inventors: Hetz Flores, Claudio Andres, Vidal Gomez, Rene Luis
• Primary Examiner(s): Kelly, Robert M
• Application Number:
  US15/773,723
• Publication Number:
  US 20190309324A1
• Time in Patent Office: 1,530 Days
• Field of Search: None
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/42 containing a HA(hemagglutinin)-tag
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector

Abstract

The present invention presents a sequence of the AAV/UPR-plus virus, a genetic treatment method and its use in the treatment of neurodegenerative diseases, such as Parkinson'"'"'s and Huntington'"'"'s diseases, among others, as presented in the in vitro studies shown in FIG. 14/17.

Claims

1. An adeno-associated vector (AAV), comprising a recombinant viral genome wherein said genome comprises an expression cassette comprising a transcription regulatory region including a promoter specific for neuronal tissues operatively bound to a polynucleotide of interest coding a fusion protein, wherein the fusion protein comprises XBP1s, ATF6f, and a bridge or linker sequence.

2. The adeno-associated vector according to claim 1, wherein the serotype of the AAV is selected from a group that comprises AAV2, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11 and pseudo-typed AAVs.

3. The adeno-associated vector according to claim 1, wherein the transcription regulatory region includes a promoter region selected from the group consisting of CMV, PGK1, CAMKII, THY1 and GAD34.

4. The adeno-associated vector according to claim 1, comprising a detection epitope coding region selected from the group consisting of Ha, Flag, Gfp, His and Myc.

5. The adeno-associated vector according to claim 1, wherein the expression cassette comprises a post-transcriptional regulatory region, and the post-transcriptional regulatory region is the post-transcriptional regulatory element of the Woodchuck Hepatitis Virus (WHP).

6. The adeno-associated vector according to claim 1, wherein the bridge sequence comprises the LGF, L4H4 and LF sequences.

7. The adeno-associated vector according to claim 1, wherein the polynucleotide of interest encodes a fusion protein selected from the group consisting of XBP1s-LGF-ATF6f-HA, XBP1s-LF-ATF6f-HA, XBP1s-L4H4-ATF6f-HA, ATF6f-LGF-XBP1s-HA, ATF6f-LF-XBP1s-HA and ATF6f-L4H4-XBP1s-HA, which act systemically close to or with neuronal cells.

8. A pharmaceutical composition comprising an adeno-associated vector of claim 1 and a pharmaceutically acceptable excipient.

9. The pharmaceutical composition according to claim 8, comprising a dosage of the virus at a range between 10⁹ to 10¹³ genome copies (GC) per ml of the pharmaceutical composition.

10. A polynucleotide, comprising an expression cassette flanked by the ITRs of an adeno-associated virus, where said expression cassette comprises a promoter, and a polynucleotide of interest that encodes a fusion protein, wherein the fusion protein comprises XBP1s, ATF6f, and a bridge or linker sequence.

11. The polynucleotide according to claim 10, wherein the promoter region is selected from the group consisting of CMV, PGK1, CAMKII, THY1 and GAD34.

12. The polynucleotide according to claim 10, wherein the expression cassette further comprises a detection epitope coding region selected from the group consisting of Ha, Flag, Gfp, His and Myc.

13. The polynucleotide according to claim 10, wherein the serotype of the adeno-associated vector is specific to neuronal cells.

14. The polynucleotide according to claim 13, wherein the serotype of the adeno-associated vector is AAV 2 or an AAV 2/6 pseudo-typed vector.

15. The polynucleotide according to claim 10, wherein the expression cassette further comprises a post-transcriptional regulatory element of the Woodchuck Hepatitis Virus (WHP).

16. The polynucleotide according to claim 10, wherein the polynucleotide of interest encodes a fusion protein selected from the group consisting of XBP1s-LGF-ATF6f-HA, XBP1s-LF-ATF6f-HA, XBP1s-L4H4-ATF6f-HA, ATF6f-LGF-XBP1s-HA, ATF6f-LF-XBP1s-HA and ATF6f-L4H4-XBP1s-HA, which act systemically close to or with neuronal cells.

18. An adeno-associated virus, comprising a viral genome comprising the polynucleotide described in claim 10.

17. A plasmid deposited in the international body of biological deposits, Instituto de Investigaciones Agropecuarias de Chile, INIA, under a deposit number selected from the group consisting of RGM 2231, RGM 2232, RGM 2233, RGM 2234, RGM 2235 and RGM 2236.