# US 10,479,821 B2 · Micro-dystrophins and related methods of use

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,479,821 B2
- Document ID: US10479821B2
- Status: Granted
- Kind: B2
- Application number: 15/541,870
- Filed: 2016-01-15
- Published: 2018-05-31
- Granted: 2019-11-19
- Earliest claimed: 2015-01-16
- Assignees: University of Washington
- Applicants: University of Washington
- Inventors: Jeffrey S. Chamberlain; Julian Ramos; Stephen D. Hauschka
- Web page: https://patentgrep.com/patent/US10479821B2

## CPC

- C07K 14/4708: {Duchenne dystrophy}
- A61K 48/0058: {Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct}
- A61P 21/00: Drugs for disorders of the muscular or neuromuscular system
- C07H 21/04: with deoxyribosyl as saccharide radical
- C07K 14/4707: {Muscular dystrophy}
- C07K 14/4716: {Muscle proteins, e.g. myosin, actin}
- C12N 15/85: for animal cells
- C12N 15/86: Viral vectors
- A61K 38/00: 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)
- C07K 2319/30: Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
- C12N 2750/14143: viral genome or elements thereof as genetic vector
- C12N 2830/008: cell type or tissue specific enhancer/promoter combination
- 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)
- 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
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Nucleotide sequences including a micro-dystrophin gene are provided. The micro-dystrophin genes may be operatively linked to a regulatory cassette. Methods of treating a subject having, or at risk of developing, muscular dystrophy, sarcopenia, heart disease, or cachexia are also provided. The methods may include administering a pharmaceutical composition including the micro-dystrophin gene and a delivery vehicle to a subject. Further, the methods may include administering the pharmaceutical composition a subject having Duchenne muscular dystrophy or Becker muscular dystrophy.

## Claims

### Claim 1

An isolated and purified nucleotide sequence, comprising:
a micro-dystrophin gene encoding a protein comprising:
an amino-terminal actin-binding domain;
a β-dystroglycan binding domain; and
a spectrin-like repeat domain, consisting of five spectrin-like repeats, including spectrin-like repeat 1 (SR1), spectrin-like repeat 16 (SR16), spectrin-like repeat 17 (SR17), spectrin-like repeat 23 (SR23), and spectrin-like repeat 24 (SR24);
wherein the micro-dystrophin gene is operatively linked to a regulatory cassette.

### Claim 2 (depends on 1)

The isolated and purified nucleotide sequence of claim 1, wherein the protein encoded by the micro-dystrophin gene further comprises at least a portion of a hinge domain.

### Claim 3 (depends on 2)

The isolated and purified nucleotide sequence of claim 2,
wherein the hinge domain is selected from at least one of a Hinge 1 domain, a Hinge 2 domain, a Hinge 3 domain, and a Hinge 4 domain.

### Claim 4 (depends on 1)

The isolated and purified nucleotide sequence of claim 1, wherein the regulatory cassette is selected from the group consisting of a CK8 promoter and a cardiac troponin T (cTnT) promoter.

### Claim 5 (depends on 1)

The isolated and purified nucleotide sequence of claim 1, wherein the regulatory cassette is a CK8 promoter, and wherein the CK8 promoter has at least 80% sequence identity to the nucleic acid sequence of SEQ ID NO:19.

### Claim 6 (depends on 1)

The isolated and purified nucleotide sequence of claim 1, wherein the regulatory cassette is a cTnT promoter, and wherein the cTnT promoter has at least 80% sequence identity to the nucleic acid sequence of SEQ ID NO:1.

### Claim 7 (depends on 1)

The isolated and purified nucleotide sequence of claim 1, wherein the micro-dystrophin gene encodes a protein comprising:
at least one of spectrin-like repeat 1 directly coupled to spectrin-like repeat 16, spectrin-like repeat 17 directly coupled to spectrin-like repeat 23, and spectrin-like repeat 23 directly coupled to spectrin-like repeat 24.

### Claim 8 (depends on 3)

The isolated and purified nucleotide sequence of claim 3, which comprises a Hinge 1 domain (H1) and a Hinge 4 domain (H4).

### Claim 9 (depends on 1)

A pharmaceutical composition, comprising:
the isolated and purified nucleotide sequence of claim 1; and
a delivery vehicle.

### Claim 10 (depends on 9)

The pharmaceutical composition of claim 9, wherein the delivery vehicle comprises an adeno-associated virus (AAV) vector or a recombinant adeno-associated virus (rAAV) vector.

### Claim 11 (depends on 10)

The delivery vehicle of claim 10, wherein the adeno-associated virus (AAV) vector consists of serotype AAV6, AAV8, or AAV9.

### Claim 12 (depends on 10)

The delivery vehicle of claim 10, wherein the recombinant adeno-associated virus (rAAV) vector consists of rAAV6, rAAV8, rAAV9, or rAAV2/6.

### Claim 13 (depends on 11)

The delivery vehicle of claim 11, wherein the adeno-associated virus (AAV) vector is serotype AAV9.

### Claim 14 (depends on 12)

The delivery vehicle of claim 12, wherein the recombinant adeno-associated virus (rAAV) vector is rAAV9.