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

Patent metadata

Field	Value
Publication number	US-9944918-B2
Application number	US-201615070787-A
Country	US
Kind code	B2
Filing date	Mar 15, 2016
Priority date	Mar 15, 2013
Publication date	Apr 17, 2018
Grant date	Apr 17, 2018

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 a nucleic acid sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6;
b) a 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, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6, and encoding a polypeptide having 100% identity to SEQ ID NO:2, and is capable of having at least equivalent expression in a host to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6 expression,
wherein the polynucleotide of a) or b) 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, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6.

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, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6.

4. The synthetic polynucleotide of claim 1 , wherein SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6 exhibits increased expression in an appropriate host relative to the expression of 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 5 , 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 the synthetic polynucleotide of claim 1 .

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

11. A method of treating a disease or condition mediated by methylmalonyl-CoA mutase, comprising administering to a cell of a subject in need thereof the polynucleotide of claim 1 , wherein the polynucleotide is inserted into the 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; and administering the cell to the subject.

Assignees

• Us Health
• The Us Secretary Dept Of Health & Human Services

Inventors

• Venditti Charles P
• Chandler Randy J

Classifications

• C12Y504/99002

  Methylmalonyl-CoA mutase (5.4.99.2) · CPC title

• C12N9/90 Primary

  Isomerases (5.) · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title