JPWO2021077000A5

JPWO2021077000A5 -

Info

Publication number: JPWO2021077000A5
Application number: JP2022522782A
Authority: JP
Publication date: 2023-10-18

Claims

a targeting moiety effective for targeting muscle cells, the targeting moiety comprising an n-mer motif; and a cargo, the cargo being attached to the targeting moiety. A composition comprising said cargos linked or otherwise associated.

the n-mer motif comprises an RGD motif or a non-RGD n-mer motif; Any one of 8692 to 8889,
A composition according to claim 1.

The RGD motif has the formula X _m RGDX _n , where m is 0 to 4 amino acids, n is 0 to 15 amino acids, and selected from the group consisting of any amino acid), or the RGD motif has the formula RGDXn, where n is 4 or 5 and X is any amino acid; 3. The composition of claim 2, wherein each X amino acid independently selected from the other from the group consisting of any amino acid.

2. The composition of claim 1, wherein the targeting moiety comprises a polypeptide, polynucleotide, lipid, polymer, sugar, viral protein, or a combination thereof.

the viral protein is a capsid protein; or the viral protein is an adeno-associated virus (AAV) protein.
The composition according to claim 4.

the n-mer motif is located between two amino acids of the viral protein such that the n-mer motif is external to the viral capsid of which the viral capsid protein is a part; or is between amino acids 262-269, 327-332, 382-386, 452-460, 488-505, 527-539, 545-558, 581-593, 704-714, or any of them in the AAV9 capsid polypeptide. or AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV rh. 74, or AAV rh. or the n-mer motif is inserted between any two consecutive amino acids at similar positions in the AAV9 capsid polypeptide or between amino acids 588 and 589 in the AAV9 capsid polypeptide or AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV rh. 74, or AAV rh. 10 inserted at a similar position in the capsid polypeptide,
The composition according to claim 4.

The composition is a modified AAV virus particle, or The composition is a modified AAV virus particle, or The composition is a modified AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV rh. 74, or AAV rh. 10 virus particles,
A composition according to claim 1.

2. The composition of claim 1, wherein the cargo is capable of treating or preventing a muscle disease or disorder.

The muscle disease or disorder is
a. autoimmune diseases;
b. cancer;
c. muscular dystrophy;
d. neuromuscular diseases;
e. Sugar or glycogen storage disease;
f. Expanded repeat disease;
g. Dominant negative disease;
h. Cardiomyopathy;
i. Viral diseases;
j. progeroid diseases; or k. 9. A composition according to claim 8, which is any combination thereof.

The cargo is
a. Morpholino;
b. Peptide-linked morpholino;
c. antisense oligonucleotide;
d. PMO, therapeutic transgene;
e. a polynucleotide encoding a therapeutic polypeptide or peptide;
f. PPMO;
g. one or more peptides or polypeptides;
h. one or more polynucleotides encoding a CRISPR-Cas protein, a guide RNA, or both;
i. a ribonucleoprotein, the ribonucleoprotein comprising a CRISPR-Cas system molecule;
j. therapeutic transgene RNA or other genetically modified or therapeutic RNA and/or proteins; or k. A composition according to claim 1, which is any combination thereof.

the cargo is capable of inducing exon skipping in a gene, or the cargo is capable of inducing exon skipping in a dystrophin gene, or the cargo is a mini- or microdystrophin gene, or The cargo is a mini- or microdystrophin gene, and the mini- or microdystrophin gene is a spectrin-like repeat 1, 1', 2, 3, 16, 17, 20, 21, 22, 23, 24, or the like. and optionally an nNOS domain, an actin binding domain, one or more hinge regions, a dystroglycan binding domain, or any combination thereof, or said cargo is operably linked to a muscle-specific promoter. has been,
A composition according to claim 1.

The stretched repeat disease is Huntington's disease, myotonic dystrophy, or facioscapulohumeral muscular dystrophy (FSHD), or the muscular dystrophy is Duchenne muscular dystrophy, Becker muscular dystrophy, limb-girdle muscular dystrophy, Emery Dreifs muscular dystrophy. , myotonic dystrophy, or FSHD, or the myotonic dystrophy is type 1 or type 2 myotonic dystrophy, or the cardiomyopathy is dilated cardiomyopathy, hypertrophic cardiomyopathy, or Duchenne muscular dystrophy. associated cardiomyopathy, or Danon's disease, or the sugar or glycogen storage disease is MPS type III disease or Pompe disease, or the MPS type III disease is MPS IIIA, IIIB, IIIC, or IIID. or the neuromuscular disease is Charcot-Marie-Tooth disease or Friedreich ataxia;
A composition according to claim 9.

2. The composition of claim 1, wherein the composition has increased muscle cell efficacy, muscle cell specificity, decreased immunogenicity, or any combination thereof.

one or more polynucleotides each encoding all or a portion of one or more targeting sites effective for targeting muscle cells, each targeting site comprising one or more n-mer motifs; wherein each n-mer motif is an RGD motif or a non-RGD n-mer motif, and each polynucleotide encodes at least one or more of said one or more n-mer motifs. a polynucleotide; and, optionally, a control element operably linked to one or more of said one or more polynucleotides.

The RGD motif has the formula X _m RGDX _n , where m is 0 to 4 amino acids, n is 0 to 15 amino acids, and selected from the group consisting of any amino acid), or the RGD motif has the formula RGDXn, where n is 4 or 5 and X is any amino acid; each X amino acid independently selected from the other from the group consisting of any amino acid), or the n-mer motif is , 6647-8313, 8314-8502, or 8692-8889,
The vector system according to claim 14.

15. The vector system of claim 14, further comprising cargo.

17. The cargo of claim 16, wherein the cargo is a cargo polynucleotide, optionally linked to one or more of the one or more polynucleotides encoding the targeting site, the control element, or both. vector system.

18. The vector system of claim 17, wherein the cargo polynucleotide is on the same vector or a different vector as the one or more polynucleotides encoding the targeting site.

said vector system is capable of producing viral particles containing cargo, or said vector system is capable of producing a viral capsid polypeptide comprising one or more of said targeting sites, or the vector system is capable of producing AAV viral particles;
The vector system according to claim 14.

The AAV virus particles include modified AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV rh. 74, or AAV rh. 10 viral particles, or the capsid polypeptide is a modified AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV rh. 74, AAV rh. 10 capsid polypeptide,
The vector system according to claim 19.

At least one of the one or more polynucleotides encoding the n-mer motif(s) comprises two amino acids of the viral protein such that at least one of the n-mer motifs is external to the viral capsid. 20. The vector system according to claim 19, wherein the vector system is inserted between two codons corresponding to .

The two codons are between amino acids 262-269, 327-332, 382-386, 452-460, 488-505, 527-539, 545-558, 581-593, 704-714 in the AAV9 capsid polypeptide, or any combination thereof or AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV rh. 74, or AAV rh. or the two codons correspond to amino acids 588 and 589 in said AAV9 capsid polynucleotide or AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7. , AAV8, AAV rh. 74, or AAV rh. 10 corresponding to similar positions in the capsid polypeptide,
22. Vector system according to claim 21.

15. The vector system of any of claims 14, wherein the vector comprising one or more polynucleotides each encoding all or a portion of the one or more targeting sites does not contain splice control elements.

15. The vector system of claim 14, further comprising a polynucleotide encoding a viral rep protein.

the viral rep protein is an AAV rep protein;
the polynucleotide encoding the viral rep protein is on the same vector or a different vector as the one or more polynucleotides each encoding all or a portion of the one or more targeting sites; or the protein is operably linked to the regulatory element;
25. Vector system according to claim 24.

A polypeptide produced by expressing a vector system according to any one of claims 14 to 25.

27. The polypeptide of claim 26, wherein the polypeptide is a viral polypeptide.

28. The polypeptide of claim 27, wherein the viral polypeptide is an AAV polypeptide.

Particles produced by expressing a vector system according to any one of claims 14 to 25.

the particle is a viral particle, or the particle is an adeno-associated virus (AAV) particle, or the particle has muscle-specific tropism.
Particles according to claim 29.

The vector system according to any one of claims 16 to 25, the polypeptide according to any one of claims 26 to 28, wherein the cargo is capable of treating or preventing a muscle disease or disorder. , or particles according to any one of claims 29-30.

The muscle disease or disorder is
a. autoimmune diseases;
b. cancer;
c. muscular dystrophy;
d. neuromuscular diseases;
e. Sugar or glycogen storage disease;
f. Expanded repeat disease;
g. Dominant negative disease;
h. Cardiomyopathy;
i. Viral diseases;
j. progeroid disease; or k. 32. The vector system, polypeptide, or particle of claim 31 in any combination thereof.

The cargo is a morpholino;
a. Peptide-linked morpholino;
b. antisense oligonucleotide;
c. PMO, therapeutic transgene;
d. a polynucleotide encoding a therapeutic polypeptide or peptide;
e. PPMO;
f. one or more peptides or polypeptides;
g. one or more polynucleotides encoding a CRISPR-Cas protein, a guide RNA, or both;
h. a ribonucleoprotein, the ribonucleoprotein comprising a CRISPR-Cas system molecule;
i. therapeutic transgene RNA or other genetically modified or therapeutic RNA and/or proteins; or j. 33. The vector system, polypeptide, or particle of claim 32 in any combination thereof.

the cargo is capable of inducing exon skipping in a gene, or the cargo is capable of inducing exon skipping in a dystrophin gene, or the cargo is a mini- or microdystrophin gene, or The cargo is a mini- or micro-dystrophin gene, and the mini- or micro-dystrophin gene comprises spectrin-like repeats 1, 1', 2, 3, 16, 17, 20, 21, 22, 23, 24, or and optionally an nNOS domain, an actin binding domain, one or more hinge regions, a dystroglycan binding domain, or any combination thereof.
32. The vector system, polypeptide, or particle of claim 31.

The stretched repeat disease is Huntington's disease, myotonic dystrophy, or facioscapulohumeral muscular dystrophy (FSHD), or the muscular dystrophy is Duchenne muscular dystrophy, Becker muscular dystrophy, limb-girdle muscular dystrophy, Emery Dreifs muscular dystrophy. , myotonic dystrophy, or FSHD, or the myotonic dystrophy is type 1 or type 2, or the cardiomyopathy is dilated cardiomyopathy, hypertrophic cardiomyopathy, DMD-related cardiomyopathy, or Danone cardiomyopathy. or the sugar or glycogen storage disease is MPS type III disease or Pompe disease, or the MPS type III disease is MPS type IIIA, IIIB, IIIC, or IIID, or the neuromuscular disease is , Charcot-Marie-Tooth disease or Friedreich ataxia,
33. The vector system, polypeptide, or particle of claim 32.

31. The polypeptide, the particle, or both have increased muscle cell potency, muscle cell specificity, decreased immunogenicity, or any combination thereof. polypeptide or particle.

a. The composition according to any one of claims 1 to 13;
b. Vector system according to any one of claims 14 to 25 or 31 to 35;
c. The polypeptide according to any one of claims 26 to 28 or 31 to 36;
d. particles according to any one of claims 29 to 36; or e. cells containing those combinations.

the cell is prokaryotic; or the cell is eukaryotic.
38. A cell according to claim 37.

a. The composition according to any one of claims 1 to 13;
b. Vector system according to any one of claims 14 to 25 or 31 to 35;
c. The polypeptide according to any one of claims 26 to 28 or 31 to 36;
d. Particles according to any one of claims 29 to 36;
e. a cell according to any one of claims 37 to 38; or f. a combination thereof; and a pharmaceutically acceptable carrier.

a. The composition according to any one of claims 1 to 13;
b. Vector system according to any one of claims 14 to 25 or 31 to 35;
c. The polypeptide according to any one of claims 26 to 28 or 31 to 36;
d. Particles according to any one of claims 29 to 36;
e. The cell according to any one of claims 37 to 38;
f. a pharmaceutical formulation according to claim 39; or g. A method comprising administering the combination to a subject in need thereof, but excluding methods of treating humans.

41. The method of claim 40, wherein the subject in need thereof has a muscular disease or disorder.

The muscle disease or disorder is
a. autoimmune diseases;
b. cancer;
c. muscular dystrophy;
d. neuromuscular diseases;
e. Sugar or glycogen storage disease;
f. Expanded repeat disease;
g. Dominant negative disease;
h. Cardiomyopathy;
i. Viral diseases;
j. progeroid diseases; or k. 42. The method of claim 41, any combination thereof.

The stretched repeat disease is Huntington's disease, myotonic dystrophy, or facioscapulohumeral muscular dystrophy (FSHD), or the muscular dystrophy is Duchenne muscular dystrophy, Becker muscular dystrophy, limb-girdle muscular dystrophy, Emery Dreifs muscular dystrophy. , myotonic dystrophy, or FSHD, or the myotonic dystrophy is type 1 or type 2, or the cardiomyopathy is dilated cardiomyopathy, hypertrophic cardiomyopathy, DMD-related cardiomyopathy, or Danone cardiomyopathy. or the sugar or glycogen storage disease is MPS type III disease or Pompe disease, or the MPS type III disease is MPS type IIIA, IIIB, IIIC, or IIID, or the neuromuscular disease is , Charcot-Marie-Tooth disease or Friedreich ataxia,
43. The method of claim 42.