# US 12,760,324 B2 · Engineered muscle targeting compositions

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- Number: US 12,760,324 B2
- Document ID: US12760324B2
- Status: Granted
- Kind: B2
- Application number: 18/915,615
- Filed: 2024-10-15
- Published: 2025-02-06
- Granted: 2026-10-13
- Earliest claimed: 2020-07-22
- Assignees: The Broad Institute, Inc.; President and Fellows of Harvard College; Massachusetts Institute of Technology
- Applicants: The Broad Institute, Inc.; President and Fellows of Harvard College; Massachusetts Institute of Technology
- Inventors: Pardis Sabeti; Mohammadsharif Tabebordbar; Simon Ye; Kim Lagerborg; Alexandra Stanton; Amy Wagers
- Web page: https://patentgrep.com/patent/US12760324B2

## CPC

- C12N 15/86: Viral vectors
- A61P 21/00: Drugs for disorders of the muscular or neuromuscular system
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- C12N 2750/14122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2750/14142: virus or viral particle as vehicle, e.g. encapsulating small organic molecule
- C12N 2750/14143: viral genome or elements thereof as genetic vector
- C12N 2750/14145: Special targeting system for viral vectors
- C12N 2750/14171: Demonstrated in vivo effect
- C12N 2830/008: cell type or tissue specific enhancer/promoter combination
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- 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)

## Abstract

Described herein are muscle-specific targeting moieties and compositions including the muscle specific targeting motifs. Also described herein are uses of the muscle-specific targeting motifs and compositions including the muscle specific targeting moieties. In some embodiments, the muscle-specific targeting moieties and compositions including the muscle specific targeting moieties can be used to direct delivery of a cargo to a muscle cell.

## Claims

### Claim 1

A composition comprising
a modified AAV capsid polypeptide comprising a targeting moiety effective at targeting a muscle cell,
wherein the targeting moiety is inserted after any one of amino acids 453, 587, and 588 of an AAV9 VP1 capsid polypeptide,
wherein the targeting moiety comprises one or more n-mer motifs,
each n-mer motif consisting of an amino acid sequence of
Xm1 RGD X1X2X3X4,
Xm2Xm1 RGD X1X2X3X4, or
Xm3Xm2Xm1 RGD X1X2X3X4,
wherein X1, X2, and X3 are independently selected from any amino acid, X4 is L or T, and
(i) Xm3 and Xm2 are independently selected from any amino acid, and Xm1 is G, S, T, N, V, R, I, K, or A, or
(ii) Xm3 and Xm1 are independently selected from any amino acid, and Xm2 is N, S, A, P, T, M, or D, or
(iii) Xm2 and Xm1 are independently selected from any amino acid, and Xm3 is T, N, S, A, G, D, or E.

### Claim 2 (depends on 1)

The composition of claim 1, wherein X4 is L, and
(I) (a) X1 is Q, H, or Y;
(b) X2 is T, G, A, or S;
(c) X3 is T, S, N, R, A;
(d) X1 is Q, H, or Y, and X2 is T, G, A, or S;
(e) X1 is Q, H, or Y, and X3 is T, S, N, R, A;
(f) X2 is T, G, A, or S, and X3 is T, S, N, R, A, or
(g) X1 is Q, H, or Y, X2 is T, G, A, or S, and X3 is T, S, N, R, or A,
or
(II) X1 is Q, X2 is T or G, and X3 is T or R; or
X1 is Q, X2 is T, and X3 is T; or
X1 is Q, X2 is G, and X3 is R,
or
(III) X1 is H, X2 is G, A, or S, and X3 is T, S, or A; or
X1 is H, X2 is G, and X3 is T or A; or
X1 is H, X2 is G, and X3 is T; or
X1 is H, X2 is G, and X3 is A,
or
(IV) X1 is H, X2 is A, and X3 is S; or
X1 is H, X2 is S, and X3 is S; or
X1 is Y, X2 is A, and X3 is N.

### Claim 3 (depends on 1)

The composition of claim 1, wherein the one or more n-mer motifs comprise
an amino acid sequence chosen from
(SEQ ID NO: 777)
GPGRGDX1X2X3X4,
(SEQ ID NO: 778)
TSVRGDX1X2X3X4,
(SEQ ID NO: 779)
AMSRGDX1X2X3X4,
(SEQ ID NO: 780)
GAVRGDX1X2X3X4,
(SEQ ID NO: 781)
TPSRGDX1X2X3X4,
(SEQ ID NO: 782)
DSRRGDX1X2X3X4,
(SEQ ID NO: 783)
NASRGDX1X2X3X4, and
(SEQ ID NO: 784)
SPVRGDX1X2X3X4
wherein X1, X2, and X3 are independently selected from any amino acid, and X4 is L.

### Claim 4 (depends on 1)

The composition of claim 1, wherein the targeting moiety further comprises one or more linkers comprising one or more glycine or glycine serine linkers.

### Claim 5 (depends on 1)

The composition of claim 1, further comprising a cargo.

### Claim 6 (depends on 5)

The composition of claim 5, wherein the cargo is
(a) capable of treating or preventing a muscle disease or disorder;
(b) a morpholino, a peptide-linked morpholino, an antisense oligonucleotide, a phosphorodiamidate morpholino oligomer (PMO), a therapeutic transgene, a polynucleotide encoding a therapeutic polypeptide or peptide, a peptide-conjugated phosphorodiamidate morpholino oligomer (PPMO), one or more peptides or polypeptides; one or more polynucleotides encoding a CRISPR-Cas protein, a guide RNA, or both, a ribonucleoprotein, wherein the ribonucleoprotein comprises a CRISPR-Cas system molecule, a therapeutic transgene RNA, or other gene-modifying or therapeutic RNA and/or protein, or any combination thereof, or
(c) both (a) and (b).

### Claim 7 (depends on 1)

A vector system comprising a polynucleotide encoding the composition of claim 1.

### Claim 8 (depends on 7)

The vector system of claim 7, further comprising a cargo, wherein the cargo is
(a) capable of treating or preventing a muscle disease or disorder;
(b) a morpholino; a peptide-linked morpholino; an antisense oligonucleotide; a PMO, a therapeutic transgene; a polynucleotide encoding a therapeutic polypeptide or peptide; a PPMO; one or more peptides or polypeptides; one or more polynucleotides encoding a CRISPR-Cas protein, a guide RNA, or both; a ribonucleoprotein, wherein the ribonucleoprotein comprises a CRISPR-Cas system molecule; a therapeutic transgene RNA, or other gene modifying or therapeutic RNA and/or protein; or any combination thereof, or
(c) both (a) and (b).

### Claim 9

An engineered particle, comprising a modified AAV capsid polypeptide comprising a targeting moiety effective at targeting a muscle cell,
wherein the targeting moiety is inserted after any one of amino acids 453, 587, and 588 of an AAV9 VP1 capsid polypeptide, or any combination thereof, or into an analogous position of an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV rh.74, or AAV rh.10 VP1 capsid polypeptide
wherein the targeting moiety comprises one or more n-mer motifs,
each n-mer motif consisting of an amino acid sequence
Xm1 RGD X1X2X3X4,
Xm2Xm1 RGD X1X2X3X4, or
Xm3Xm2Xm1 RGD X1X2X3X4,
wherein X1, X2, and X3 are independently selected from any amino acid, X4 is L or T, and
(i) Xm3 and Xm2 are independently selected from any amino acid, and Xm1 is G, S, T, N, V, R, I, K, or A, or
(ii) Xm3 and Xm1 are independently selected from any amino acid, and Xm2 is N, S, A, P, T, M, or D, or
(iii) Xm2 and Xm1 are independently selected from any amino acid, and Xm3 is T, N, S, A, G, D, or E.

### Claim 10 (depends on 9)

The engineered particle of claim 9, wherein the targeting moiety further comprises one or more linkers comprising one or more glycine or glycine serine linkers.

### Claim 11 (depends on 9)

The engineered particle of claim 9, further comprising a cargo.

### Claim 12 (depends on 11)

The engineered particle of claim 11, wherein the cargo is
(a) capable of treating or preventing a muscle disease or disorder;
(b) a morpholino, a peptide-linked morpholino, an antisense oligonucleotide, a PMO, a therapeutic transgene, a polynucleotide encoding a therapeutic polypeptide or peptide, a PPMO, one or more peptides or polypeptides; one or more polynucleotides encoding a CRISPR-Cas protein, a guide RNA, or both, a ribonucleoprotein, wherein the ribonucleoprotein comprises a CRISPR-Cas system molecule, a therapeutic transgene RNA, or other gene-modifying or therapeutic RNA and/or protein, or any combination thereof, or
(c) both (a) and (b).