Synthetic methylmalonyl-CoA mutase transgene for the treatment of MUT class methylmalonic acidemia (MMA)

Patent metadata

Field	Value
Publication number	US-9719080-B2
Application number	US-201414773885-A
Country	US
Kind code	B2
Filing date	Mar 14, 2014
Priority date	Mar 15, 2013
Publication date	Aug 1, 2017
Grant date	Aug 1, 2017

Abstract

Synthetic polynucleotides encoding human methylmalonyl-CoA mutase (synMUT) and exhibiting augmented expression in cell culture and/or in a subject are described herein. An adeno-associated viral (AAV) gene therapy vector encoding synMUT under the control of a liver-specific promoter (AAV2/8-HCR-hAAT-synMUT-RBG) successfully rescued the neonatal lethal phenotype displayed by methylmalonyl-CoA mutase-deficient mice, lowered circulating methylmalonic acid levels in the treated animals, and resulted in prolonged hepatic expression of the product of synMUT transgene in vivo, human methylmalonyl-CoA mutase (MUT).

First claim

The invention claimed is:

1. A synthetic methylmalonyl-CoA mutase (MUT) polynucleotide (synMUT) selected from the group consisting of:
(a) a polynucleotide comprising the nucleic acid sequence of SEQ ID NO:1; and
(b) a codon-optimized polynucleotide comprising a polynucleotide having a nucleic acid sequence with at least about 80% identity to the nucleic acid sequence of SEQ ID NO:1 and encoding a polypeptide according to SEQ ID NO:2, and having equivalent expression in a host to either SEQ ID NO:1 expression or SEQ ID NO:3 expression,
wherein the codon-optimized polynucleotide having at least about 80% identity to SEQ ID NO:1 does not have the nucleic acid sequence of SEQ ID NO:3.

2. The synthetic polynucleotide of claim 1 , wherein the polynucleotide has at least about 90% identity to the nucleic acid sequence of SEQ ID NO:1.

3. The synthetic polynucleotide of claim 1 , wherein the polynucleotide has at least about 95% identity to the nucleic acid sequence of SEQ ID NO:1.

4. The synthetic polynucleotide of claim 1 , wherein the SEQ ID NO:1 exhibits increased expression in an appropriate host relative to the expression SEQ ID NO:3 in an appropriate host.

5. The synthetic polynucleotide of claim 3 , wherein the synthetic polynucleotide having increased expression comprises a nucleic acid sequence comprising codons that have been optimized relative to the naturally occurring human methylmalonyl-CoA mutase polynucleotide sequence (SEQ ID NO:3).

6. The synthetic polynucleotide of claim 4 , wherein the nucleic acid sequence has at least about 70% of less commonly used codons replaced with more commonly used codons.

7. An expression vector comprising the synthetic polynucleotide of claim 1 .

8. The expression vector of claim 7 , wherein the expression vector is AAV2/8-HCR-hAAT-RBG.

9. A method of treating a disease or condition mediated by methylmalonyl-CoA mutase, comprising administering to a subject in need thereof a therapeutic amount of a synthetic methylmalonyl-CoA mutase (MUT) polynucleotide (synMUT) selected from the group consisting of:
a) a polynucleotide comprising the nucleic acid sequence of SEQ ID NO:1;
b) a codon optimized polynucleotide comprising a polynucleotide having a nucleic acid sequence with at least about 80% identity to the nucleic acid sequence of SEQ ID NO:1 and encoding a polypeptide according to SEQ ID NO:2, and having equivalent expression in a host to either SEQ ID NO:1 expression or SEQ ID NO:3 expression,
wherein the codon optimized polynucleotide having at least about 80% identity to SEQ ID NO:1 does not have the nucleic acid sequence of SEQ ID NO:3.

10. The method of claim 9 , wherein the disease or condition is methylmalonic academia (MMA).

11. The synthetic polynucleotide of claim 1 , wherein the synthetic polynucleotide is formulated with a pharmaceutically acceptable carrier.

12. The method of claim 9 , wherein the polynucleotide is inserted into a cell of the subject via genome editing on the cell of the subject using a nuclease selected from the group of zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), the clustered regularly interspaced short palindromic repeats (CRISPER/cas system) and meganuclease re-engineered homing endonucleases on a cell from the subject.

13. The synthetic polynucleotide of claim 1 , wherein the polynucleotide has at least about 97% identity to the nucleic acid sequence of SEQ ID NO:1.

14. The synthetic polynucleotide of claim 1 , wherein the polynucleotide has at least about 99% identity to the nucleic acid sequence of SEQ ID NO:1.

15. The method of claim 9 , wherein the polynucleotide has at least about 90% identity to the nucleic acid sequence of SEQ ID NO:1.

16. The method of claim 9 , wherein the polynucleotide has at least about 95% identity to the nucleic acid sequence of SEQ ID NO:1.

17. The method of claim 9 , wherein the polynucleotide has at least about 97% identity to the nucleic acid sequence of SEQ ID NO:1.

18. The method of claim 9 , wherein the polynucleotide has at least about 99% identity to the nucleic acid sequence of SEQ ID NO:1.

Assignees

• Us Health
• Us Health

Inventors

• Venditti Charles P
• Chandler Randy J

Classifications

• A61K48/0066

  Manipulation of the nucleic acid to modify its expression pattern, e.g. enhance its duration of expression, achieved by the presence of particular introns in the delivered nucleic acid · CPC title

• A61K38/52 Primary

  Isomerases (5) · CPC title

• C12Y504/99002

  Methylmalonyl-CoA mutase (5.4.99.2) · CPC title

• C12N9/90 Primary

  Isomerases (5.) · CPC title