Compositions and methods for internalizing enzymes

Patent metadata

Field	Value
Publication number	US-2025171520-A1
Application number	US-202418973895-A
Country	US
Kind code	A1
Filing date	Dec 9, 2024
Priority date	Jun 7, 2017
Publication date	May 29, 2025
Grant date	—

Abstract

Compositions and methods for treating enzyme-deficiency diseases are disclosed. Multidomain therapeutic proteins containing an internalization effector binding domain and a lysosomal replacement enzyme activity are disclosed. The multidomain therapeutic proteins are capable of entering cells, segregating to the lysosome, and delivering the replacement enzyme activity to the lysosome.

First claim

1 .- 31 . (canceled)

32 . A method of reducing glycogen accumulation in a tissue in a subject in need thereof, comprising administering to the subject a polynucleotide encoding a multidomain therapeutic protein comprising a delivery domain and an enzyme domain,
wherein the delivery domain is an antigen-binding protein that binds to CD63, wherein the enzyme domain comprises an alpha-glucosidase (GAA) or a portion thereof, and wherein the polynucleotide further comprises a virus nucleic acid sequence, wherein the virus nucleic acid sequence is an adeno-associated virus (AAV) nucleic acid sequence.

33 . The method of claim 32 , wherein the antigen-binding protein is a single-chain variable fragment (scFv).

34 . The method of claim 32 , wherein the tissue is selected from the group consisting of heart, liver, and skeletal muscle.

35 . The method of claim 32 , wherein
(i) high serum levels of GAA in the subject are maintained for at least 12 weeks; and/or (ii) glycogen levels in the tissue are maintained at wild-type levels 3 months after administration of the polynucleotide.

36 . The method of claim 32 , wherein the polynucleotide comprises a locus-targeting nucleic acid sequence.

37 . The method of claim 32 , wherein the polynucleotide comprises a tissue-specific regulatory element.

38 . The method of claim 37 , wherein the tissue-specific regulatory element is a liver-specific promoter.

39 . The method of claim 32 , wherein the enzyme domain comprises the amino acid sequence set forth in SEQ ID NO: 1 or a biologically active portion thereof.

40 . The method of claim 32 , wherein the delivery domain comprises:
the amino acid sequence set forth in SEQ ID NO: 2; the HCDR-1-HCDR-2-HCDR3-LCDR1-LCDR2-LCDR-3 amino acid sequences contained within an HCVR/LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 14/22, SEQ ID NOS: 30/38, SEQ ID NOS: 46/54, and SEQ ID NOS: 62/70; HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences selected from the group consisting of SEQ ID NOs: 16-18-20-24-26-28, SEQ ID NOs: 32-34-36-40-42-44, SEQ ID NOs: 48-50-52-56-58-60, and SEQ ID NOs: 64-66-68-72-74-76; or an HCVR/LCVR amino acid pair set forth in SEQ ID NOS 14/22, SEQ ID NOS: 30/38, SEQ ID NOS: 46/54, or SEQ ID NOS: 62/70.

41 . The method of claim 40 , wherein the HCVR/LCVR amino acid pair comprises from N-terminal to C-terminal: the HCVR, a linker, and the LCVR.

42 . The method of claim 41 , wherein the enzyme domain is attached to the C-terminus of the LCVR via a linker.

43 . The method of claim 42 , wherein the delivery domain comprises the HCVR/LCVR amino acid pair set forth in SEQ ID NOS: 46/54.

44 . The method of claim 32 , wherein the multidomain therapeutic protein comprises the amino acid sequence set forth in SEQ ID NO: 79.

45 . The method of claim 32 , wherein the polynucleotide is administered via a gene therapy vector comprising the polynucleotide, and wherein the gene therapy vector is an AAV vector.

46 . The method of claim 45 , wherein the gene therapy vector is an AAV8 vector or an AAV2/8 chimera.

47 . The method of claim 45 , wherein the gene therapy vector is in a composition further comprising a pharmaceutically acceptable carrier.

48 . The method of claim 45 , wherein the method results in the polynucleotide being integrated into a genomic locus of one or more cells of the subject.

49 . The method of claim 48 , wherein the subject is a human, and the one or more cells are human cells.

50 . The method of claim 48 , wherein the genomic locus is a safe harbor locus selected from the group consisting of an EESYR locus, a SARS locus, position 188,083,272 of human chromosome 1 or its non-human mammalian orthologue, position 3,046,320 of human chromosome 10 or its non-human mammalian orthologue, position 67,328,980 of human chromosome 17 or its non-human mammalian orthologue, an adeno-associated virus site 1 (AAVS1) on human chromosome 19 or its non-human mammalian orthologue, a chemokine receptor 5 (CCR5) gene, a chemokine receptor gene encoding an HIV-1 coreceptor, a mouse Rosa26 locus or its non-murine mammalian orthologue, and a human albumin (alb) locus.

Assignees

• Regeneron Pharma

Inventors

• Baik Andrew
• Cygnar Katherine
• Schoenherr Christopher
• Kyratsous Christos
• Wang Cheng

Classifications

• C07K2319/33

  fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• A61K38/47 Primary

  acting on glycosyl compounds (3.2), e.g. cellulases, lactases · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• C07K2317/24

  containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered · CPC title