Compositions useful in treatment of ornithine transcarbamylase (OTC) deficiency

Patent metadata

Field	Value
Publication number	US-11732246-B2
Application number	US-202016995572-A
Country	US
Kind code	B2
Filing date	Aug 17, 2020
Priority date	Mar 9, 2014
Publication date	Aug 22, 2023
Grant date	Aug 22, 2023

Abstract

Viral vectors comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis and/or cirrhosis in OTCD patients by administering hOTC.

First claim

What is claimed is:

1. A nucleic acid molecule comprising a sequence encoding a functional human ornithine transcarbamylase (hOTC) protein,
wherein the sequence is less than 80% identical to the wild-type hOTC sequence of SEQ ID NO: 1, and
wherein said sequence is selected from a nucleic acid sequence comprising SEQ ID NO: 5 or a nucleic acid sequence at least 96% identical thereto.

2. The nucleic acid molecule according to claim 1 , wherein the sequence encoding the functional hOTC protein has the sequence of SEQ ID NO: 5.

3. The nucleic acid molecule according to claim 1 , wherein the sequence encoding the functional hOTC protein has the sequence of SEQ ID NO: 3 or of SEQ ID NO: 4.

4. A plasmid containing an expression cassette comprising:
an AAV 5′ inverted terminal repeat (ITR),
an hOTC nucleic acid sequence,
expression control sequences,
and an AAV 3′ ITR,
wherein the hOTC nucleic acid sequence encodes a functional hOTC protein and is less than 80% identical to the wild-type hOTC sequence of SEQ ID NO: 1, and
wherein said hOTC nucleic acid sequence is selected from a nucleic acid sequence comprising SEQ ID NO: 5 or a nucleic acid sequence at least 96% identical thereto.

5. The plasmid according to claim 4 , wherein the hOTC nucleic acid sequence has the sequence of SEQ ID NO: 5.

6. The plasmid according to claim 4 , wherein the hOTC nucleic acid sequence has the sequence of SEQ ID NO: 3 or of SEQ ID NO: 4.

7. The plasmid according to claim 4 , wherein the expression control sequences comprise a liver-specific promoter.

8. The plasmid according to claim 7 , wherein the liver-specific promoter is a thyroxin-binding globulin (TBG) promoter.

9. The plasmid according to claim 4 , wherein the expression cassette further comprises one or more of an intron, a Kozak sequence, a poly A, and a post-transcriptional regulatory element.

10. The plasmid according to claim 4 , wherein the AAV 5′ ITR and/or the AAV 3′ ITR is/are from AAV2.

11. The plasmid according to claim 4 , wherein the expression cassette comprises one or more enhancers.

12. The plasmid according to claim 11 , wherein the enhancers include an alpha 1 microglobulin/bikunin (alpha mic/bik) enhancer.

13. A producer cell containing the plasmid according to claim 4 .

14. A plasmid comprising a vector genome which comprises
an AAV2 3′ ITR,
an AAV2 5′ ITR in which the D-sequence and terminal resolution site (trs) are deleted,
a thyroxin-binding globulin (TBG) promoter sequence,
an hOTC coding sequence comprising SEQ ID NO: 5, and
an SV40 polyA sequence.

15. A producer cell containing the plasmid according to claim 14 .

16. A plasmid containing an expression cassette comprising:
an AAV 5′ ITR,
a nucleic acid sequence that encodes a chimeric ornithine transcarbamylase comprising a mature hOTC protein and a heterologous transit peptide,
expression control sequences,
and an AAV 3′ ITR,
wherein the mature hOTC protein is encoded by a sequence less than 80% identical to the wild-type mature hOTC coding sequence of SEQ ID NO: 1, and
wherein the mature hOTC protein is encoded by a nucleic acid sequence comprising the mature hOTC coding sequence of SEQ ID NO: 5 or a nucleic acid sequence at least 96% identical thereto.

17. The plasmid according to claim 16 , wherein the nucleic acid sequence that encodes the chimeric ornithine transcarbamylase comprises nucleotides 97 to 1062 of SEQ ID NO: 5.

18. The plasmid according to claim 16 , wherein the expression control sequences comprise a liver-specific promoter.

19. The plasmid according to claim 16 , wherein the expression cassette further comprises one or more of an intron, a Kozak sequence, a poly A, and a post-transcriptional regulatory element.

20. A producer cell containing the plasmid according to claim 16 .

21. A packaging cell line containing the plasmid according to claim 16 .

22. A packaging cell line containing the plasmid according to claim 4 .

23. A packaging cell line containing the plasmid according to claim 14 .

Assignees

• Univ Pennsylvania

Inventors

• Wang Lili
• Wilson James M

Classifications

• C12N9/1018 Primary

  Carboxy- and carbamoyl transferases (2.1.3) · CPC title

• A61K48/0058

  Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct · CPC title

• C12N15/86 Primary

  Viral vectors · CPC title

• C12Y201/03003

  Ornithine carbamoyltransferase (2.1.3.3) · CPC title

• C12N2710/10343

  viral genome or elements thereof as genetic vector · CPC title