# US 11,298,429 B2 · Adeno-associated virus vector delivery of microrna-29 to treat muscular dystrophy

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- Number: US 11,298,429 B2
- Document ID: US11298429B2
- Status: Granted
- Kind: B2
- Application number: 16/092,711
- Filed: 2017-04-14
- Published: 2019-04-25
- Granted: 2022-04-12
- Earliest claimed: 2016-04-15
- Assignees: Research Institute At Nationwide Children's Hospital
- Applicants: Research Institute At Nationwide Children's Hospital
- Inventors: Louise Rodino-Klapac; Jerry R. Mendell; Kristin N. Heller
- Web page: https://patentgrep.com/patent/US11298429B2

## CPC

- A61K 48/005: {characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered}
- A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)}
- A61K 31/7088: Compounds having three or more nucleosides or nucleotides
- A61K 35/761: Adenovirus
- A61K 38/1709: {from mammals}
- A61K 38/1719: {Muscle proteins, e.g. myosin or actin}
- A61K 48/0058: {Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct}
- A61K 48/0075: {characterised by an aspect of the delivery route, e.g. oral, subcutaneous}
- A61P 19/04: for non-specific disorders of the connective tissue
- A61P 21/00: Drugs for disorders of the muscular or neuromuscular system
- C07K 14/4708: {Duchenne dystrophy}
- C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids}
- C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)}
- C12N 15/86: Viral vectors
- C12N 15/8645: {Adeno-associated virus}
- C12N 2310/141: MicroRNAs, miRNAs
- C12N 2320/31: Combination therapy
- C12N 2330/51: Specially adapted vectors
- C12N 2750/14141: Use of virus, viral particle or viral elements as a vector
- C12N 2750/14143: viral genome or elements thereof as genetic vector
- C12N 2800/22: Vectors comprising a coding region that has been codon optimised for expression in a respective host
- C12N 2830/008: cell type or tissue specific enhancer/promoter combination
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The invention provides for recombinant AAV vectors comprising a polynucleotide sequence comprising the guide strand of miR-29c and methods of using the recombinant vectors to reduce or prevent fibrosis in subjects suffering from muscular dystrophy.

## Claims

### Claim 1

A recombinant AAVrh74 vector comprising the nucleic sequences of SEQ ID NO: 3 or SEQ ID NO: 4.

### Claim 2 (depends on 1)

The recombinant AAVrh74 vector of claim 1 comprising
a) the nucleotide sequence of SEQ ID NO: 2,
b) the nucleotide sequence of SEQ ID NO: 1 or
c) the nucleotide sequence of SEQ ID NO: 12.

### Claim 3 (depends on 1)

The recombinant AAV vector of claim 1 wherein the polynucleotide sequence is operably linked to a muscle-specific control element.

### Claim 4 (depends on 3)

The recombinant AAV vector of claim 3 wherein the muscle-specific control element is cytomegalovirus (CMV) promoter, human skeletal actin gene element, cardiac actin gene element, myocyte-specific enhancer binding factor mef, muscle creatine kinase (MCK), truncated MCK (tMCK), myosin heavy chain (MHC), MHCK7, C5-12, murine creatine kinase enhancer element, skeletal fast-twitch troponin c gene element, slow-twitch cardiac troponin c gene element, the slow-twitch troponin i gene element, hypoxia-inducible nuclear factors, steroid-inducible element or glucocorticoid response element (gre).

### Claim 5 (depends on 3)

The recombinant AAV vector of claim 3 wherein the muscle-specific control element comprises the nucleotide sequence of SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 13.

### Claim 6 (depends on 1)

A composition comprising the recombinant AAV vector of claim 1.

### Claim 7 (depends on 6)

A method of treating muscular dystrophy comprising administering a therapeutically effective amount of the composition of claim 6.

### Claim 8 (depends on 6)

A method of reducing or preventing fibrosis in a subject suffering from muscular dystrophy comprising administering a therapeutically effective amount of the composition of claim 6.

### Claim 9 (depends on 6)

A method of preventing fibrosis in a patient suffering from muscular dystrophy comprising administering a therapeutically effective amount of the composition of claim 6.

### Claim 10 (depends on 6)

A method of increasing muscular force or muscle mass in a subject suffering from muscular dystrophy comprising administering a therapeutically effective amount of the composition of claim 6.

### Claim 11 (depends on 7)

The method claim 7 wherein the muscular dystrophy is Duchenne muscular dystrophy or Limb Girdle muscular dystrophy.

### Claim 12 (depends on 7)

The method of claim 7 wherein the recombinant AAV vector or the composition is administered by intramuscular injection.

### Claim 13 (depends on 7)

The method of claim 7 wherein the recombinant AAV vector or the composition is administered systemically.

### Claim 14 (depends on 13)

The method of claim 13, where the recombinant AAV vector or the composition is parenterally administered by injection, infusion or implantation.

### Claim 15 (depends on 7)

The method of claim 7, wherein the recombinant AAV vector of composition is administered before fibrosis is observed in the subject or before muscle force is reduced in the subject or before muscle mass is reduced in the subject.

### Claim 16 (depends on 1)

A method of producing a mature microRNA polynucleotide sequence comprising infecting a host cell with a recombinant AAV vector claim 1 and expressing a mature microRNA polynucleotide in the host cell.

### Claim 17 (depends on 2)

The recombinant AAV vector of claim 2 wherein the polynucleotide sequence is operably linked to a muscle-specific control element.

### Claim 18 (depends on 17)

The recombinant AAV vector of claim 17 wherein the muscle-specific control element is cytomegalovirus (CMV) promoter, human skeletal actin gene element, cardiac actin gene element, myocyte-specific enhancer binding factor mef, muscle creatine kinase (MCK), truncated MCK (tMCK), myosin heavy chain (MHC), MHCK7, C5-12, murine creatine kinase enhancer element, skeletal fast-twitch troponin c gene element, slow-twitch cardiac troponin c gene element, the slow-twitch troponin i gene element, hypoxia-inducible nuclear factors, steroid-inducible element or glucocorticoid response element (gre).

### Claim 19 (depends on 17)

The recombinant AAV vector of claim 17 wherein the muscle-specific control element comprises the nucleotide sequence of SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 13.

### Claim 20 (depends on 2)

A composition comprising the recombinant AAV vector of claim 2.