Granted Patent B2

US 12,668,813 · App. 17/487,699 · Granted Jun 30, 2026

Inventor: Jie Xu (State College, PA)

Assignee: NeuExcell Therapeutics Inc.

C12N15/86A61K35/761

Abstract

Claims (41)

1 . An adeno-associated virus (AAV) vector comprising a human neurogenic differentiation 1 (hNeuroD1) sequence, wherein said hNeuroD1 sequence comprises a nucleic acid sequence at least 99% identical to SEQ ID NO:6; wherein the AAV vector does not encode another heterologous polypeptide; and wherein said hNeuroD1 sequence is operably linked to regulatory elements comprising:

(a) a glial fibrillary acidic protein (GFAP) promoter comprising a nucleic acid sequence at least 95% identical to the sequence set forth in SEQ ID NO:15;

(b) a cytomegalovirus (CMV) enhancer comprising a nucleic acid sequence at least 95% identical to the sequence set forth in SEQ ID NO:11;

(c) a chimeric intron comprising a nucleic acid sequence at least 95% identical to the sequence set forth in SEQ ID NO:16;

(d) a woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) comprising the nucleic acid sequence set forth in SEQ ID NO:18; and

(e) a bGH polyadenylation sequence comprising a nucleic acid sequence at least 95% identical to the sequence set forth in SEQ ID NO:14.

2 . The AAV vector of claim 1 , wherein:

(a) said GFAP promoter comprises the nucleic acid sequence set forth in SEQ ID NO: 15;

(b) said CMV enhancer comprises the nucleic acid sequence set forth in SEQ ID NO: 11;

(c) said chimeric intron comprises the nucleic acid sequence set forth in SEQ ID NO: 16;

(d) said bGH polyadenylation sequence comprises the nucleic acid sequence set forth in SEQ ID NO:14; or

(e) said nucleic acid sequence encoding said hNeuroD1 sequence is at least 99.5% identical to the sequence set forth in SEQ ID NO:6.

3 . The AAV vector of claim 1 , wherein said AAV vector is selected from the group consisting of AAV serotype 2, AAV serotype 5, and AAV serotype 9.

4 . The AAV vector of claim 1 , wherein said hNeuroD1 sequence comprises the nucleic acid sequence set forth in SEQ ID NO:6.

5 . The AAV vector of claim 1 , wherein said AAV vector further comprises a 2A self-cleavage peptide coding sequence that is at least 80% identical to (i) the sequence set forth in SEQ ID NO:3 or (ii) the complementary sequence set forth in SEQ ID NO:3.

6 . The AAV vector of claim 1 , wherein said AAV vector comprises at least one inverted terminal repeat nucleic acid sequence at least 80% identical to the sequence set forth in SEQ ID NO: 1 or SEQ ID NO:9.

7 . An adeno-associated virus (AAV) vector comprising, in order from 5′ to 3′:

(a) a cytomegalovirus (CMV) enhancer comprising a nucleic acid sequence at least 95% identical to the sequence set forth in SEQ ID NO:11;

(b) a glial fibrillary acidic protein (GFAP) promoter comprising a nucleic acid sequence at least 95% identical to the sequence set forth in SEQ ID NO:15;

(c) a chimeric intron comprising a nucleic acid sequence at least 95% identical to the sequence set forth in SEQ ID NO:16;

(d) a human neurogenic differentiation 1 (hNeuroD1) sequence comprising a nucleic acid sequence at least 99% identical to the sequence set forth in SEQ ID NO:6;

(e) a woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) comprising the nucleic acid sequence set forth in SEQ ID NO:18; and

(f) a bGH polyadenylation sequence comprising a nucleic acid sequence at least 95% identical to the sequence set forth in SEQ ID NO:14;

wherein the AAV vector does not encode another heterologous polypeptide.

8 . The AAV vector of claim 7 , wherein said AAV vector comprises at least one inverted terminal repeat nucleic acid sequence at least 80% identical to the sequence set forth in SEQ ID NO:1 or SEQ ID NO:9.

9 . The AAV vector of claim 7 , wherein:

(a) said GFAP promoter comprises the nucleic acid sequence set forth in SEQ ID NO: 15;

(b) said CMV enhancer comprises the nucleic acid sequence set forth in SEQ ID NO: 11;

(c) said chimeric intron comprises the nucleic acid sequence set forth in SEQ ID NO: 16;

(d) said bGH polyadenylation sequence comprises the nucleic acid sequence set forth in SEQ ID NO:14; and

(e) said hNeuroD1 sequence comprises a nucleic acid sequence at least 99.5% identical to the sequence set forth in SEQ ID NO:6.

10 . The AAV vector of claim 1 , wherein said AAV vector further comprises a 2A self-cleavage peptide coding sequence at least 90% identical to (i) the sequence set forth in SEQ ID NO:3 or (ii) the complementary sequence set forth in SEQ ID NO:3.

11 . The AAV vector of claim 1 , wherein said AAV vector further comprises a 2A self-cleavage peptide coding sequence comprising the nucleic acid sequence set forth in SEQ ID NO:3 or the complementary sequence set forth in SEQ ID NO:3.

12 . The AAV vector of claim 1 , wherein said AAV vector comprises at least one inverted terminal repeat nucleic acid sequence at least 90% identical to the sequence set forth in SEQ ID NO:1 or SEQ ID NO:9.

13 . The AAV vector of claim 1 , wherein said AAV vector comprises at least one inverted terminal repeat nucleic acid sequence set forth in SEQ ID NO:1 or SEQ ID NO:9.

14 . The AAV vector of claim 7 , wherein said AAV vector is selected from the group consisting of AAV serotype 2, AAV serotype 5, and AAV serotype 9.

15 . The AAV vector of claim 7 , wherein said AAV vector further comprises a 2A self-cleavage peptide coding sequence at least 90% identical to (i) the sequence set forth in SEQ ID NO:3 or (ii) the complementary sequence set forth in SEQ ID NO:3.

16 . The AAV vector of claim 7 , wherein said AAV vector further comprises a 2A self-cleavage peptide coding sequence comprising the nucleic acid sequence set forth in SEQ ID NO:3 or the complementary sequence set forth in SEQ ID NO:3.

17 . The AAV vector of claim 7 , wherein said AAV vector comprises at least one inverted terminal repeat nucleic acid sequence at least 90% identical to the sequence set forth in SEQ ID NO: 1 or SEQ ID NO: 9.

18 . The AAV vector of claim 7 , wherein said AAV vector comprises at least one inverted terminal repeat nucleic acid sequence set forth in SEQ ID NO: 1 or SEQ ID NO:9.

19 . The AAV vector of claim 1 , wherein said hNeuroD1 sequence comprises the amino acid sequence set forth in SEQ ID NO:10.

Assignments (2)

Continuity (3)

References Cited (400)