# US 2020/0224205 A1 · Vlp for the treatment of a lysosomal storage disease

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

- Number: US 2020/0224205 A1
- Document ID: US20200224205A1
- Status: Published application
- Kind: A1
- Application number: 16/717,033
- Filed: 2019-12-17
- Published: 2020-07-16
- Earliest claimed: 2018-12-18
- Assignees: NEUWAY Pharma GmbH
- Applicants: NEUWAY Pharma GmbH
- Inventors: Victoria DEMINA; Marcus STAPF; Sergey Sotnikov; Heiko MANNINGA
- Web page: https://patentgrep.com/patent/US20200224205A1

## CPC

- C12N 15/63: Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- A61K 9/5184: {Virus capsids or envelopes enclosing drugs (liposomes with additional exogenous lipids A61K9/127)}
- C07K 14/005: from viruses
- C12N 9/16: acting on ester bonds (3.1)
- C12N 15/52: Genes encoding for enzymes or proenzymes
- C12N 15/86: Viral vectors
- C12N 2710/22022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2710/22023: Virus like particles [VLP]
- C12N 2710/22032: Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
- C12N 2710/22042: virus or viral particle as vehicle, e.g. encapsulating small organic molecule
- C12N 2710/22051: Methods of production or purification of viral material
- C12Y 301/06001: Arylsulfatase (3.1.6.1)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)
- 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)
- C12Y: ENZYMES

## Abstract

The invention relates to virus like particles (VLP) associated with a lysosomal enzyme or an expression vector encoding a lysosomal enzyme which are used in a method for the treatment of a lysosomal storage disease. The invention also relates to a pharmaceutical composition for use in a method for the treatment of a lysosomal storage disease, to an expression vector encoding a lysosomal enzyme and to a method of associating a VLP with an expression vector encoding a lysosomal enzyme.

## Claims

### Claim 1

A method of treating Metachromatic Leukodystrophy Disease (MLD), comprising administering a virus-like particle (VLP) to a subject in need thereof,
wherein said VLP incorporates a lysosomal enzyme, or incorporates an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, and
wherein the VLP is derived from JCV.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the VLP does not comprise viral genetic material, and the expression vector does not encode viral proteins.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the subject is an animal.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the subject has a neurologic deficit.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the lysosomal enzyme is a human enzyme.

### Claim 6 (depends on 1)

The method according to claim 1, wherein the lysosomal enzyme is arylsulfatase A.

### Claim 7 (depends on 1)

The method according to claim 1, wherein the lysosomal enzyme comprises an amino acid sequence which is at least 80% identical to the amino acid sequence of SED NO: 1 over its entire length.

### Claim 8 (depends on 1)

The method according to claim 1, wherein the VLP incorporates an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, and wherein the expression vector has a size of less than 7 kb.

### Claim 9 (depends on 1)

The method according to claim 1, wherein the VLP incorporates an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, and wherein the expression vector has a promoter selected from the group consisting of a CMV and a CAG promoter.

### Claim 10 (depends on 1)

The method according to claim 1, wherein the VLP incorporates an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, and wherein the nucleotide sequence encoding the lysosomal enzyme is at least 70% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length.

### Claim 11 (depends on 1)

The method according to claim 1, wherein after said administration, the VLP crosses the blood-brain barrier to enter the CNS together with the lysosomal enzyme or the expression vector.

### Claim 12 (depends on 11)

The method according to claim 11, wherein the lysosomal enzyme or the expression vector enters at least one cell type selected from the group consisting of astrocytes, oligodendrocytes, microglia and neurons, after crossing said blood-brain barrier.

### Claim 13 (depends on 1)

The method according to claim 1, wherein the VLP is administered orally or parenterally.

### Claim 14 (depends on 1)

The method according to claim 1, wherein after said administration, a target cell is contacted with an effective amount of the lysosomal enzyme.

### Claim 15 (depends on 1)

The method according to claim 1, wherein the lysosomal enzyme has a therapeutically effective enzyme activity for at least 10 days.

### Claim 16 (depends on 1)

The method according to claim 1, wherein the VLP is composed of VP1 proteins of JC virus.

### Claim 17 (depends on 16)

The method according to claim 16, wherein the VP1 protein comprises an amino acid sequence which is at least 80% identical to the amino acid sequence of SEQ ID NO: 3 over its entire length.

### Claim 18 (depends on 1)

The method according to claim 1, wherein said VLP is administered in a pharmaceutical composition that further comprises at least one of a pharmaceutically acceptable carrier and an excipient.

### Claim 19

An expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, and further comprising a promoter selected from the group consisting of CAG and CMV, wherein said expression vector has a size of less than 7 kb.

### Claim 20 (depends on 19)

The expression vector according to claim 19, wherein the nucleotide sequence encoding the lysosomal enzyme is at least 70% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length.

### Claim 21

A method of incorporating a lysosomal enzyme, or incorporating an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, into a VLP, wherein the method comprises:
a) providing a composition comprising VP1 proteins,
b) exposing the VP1 proteins of the composition of a) to conditions inducing the VP1 to assemble into VLP,
c) exposing the VLP of the composition of b) to conditions disassembling the VLP into pentamers,
d) exposing the pentamers of the composition of c) to conditions inducing the pentamers to reassemble into VLP,
e) exposing the VLP of the composition of d) to conditions disassembling the VLP into pentamers, and
f) exposing the pentamers of the composition of e) to the lysosomal enzyme or the expression vector to conditions inducing the pentamers to assemble into a VLP associated with the lysosomal enzyme or the expression vector.

### Claim 22 (depends on 21)

A VLP produced by the method according to claim 21.

### Claim 23 (depends on 1)

A VLP incorporating a lysosomal enzyme, or incorporating an expression vector comprising a nucleotide sequence encoding a lysosomal enzyme, for use in the method of claim 1.

### Claim 24 (depends on 23)

The VLP according to claim 23, wherein the VLP does not comprise viral genetic material, and the expression vector does not encode viral proteins.

### Claim 25 (depends on 1)

The method according to claim 1, wherein said method does not comprise a step of increasing the permeability of a blood-brain barrier of the subject.

### Claim 26 (depends on 23)

The VLP according to claim 23, wherein the VLP is derived from JCV.

### Claim 27 (depends on 23)

The VLP according to claim 23, wherein the lysosomal enzyme is arylsulfatase A.

### Claim 28 (depends on 23)

The VLP according to claim 23, wherein the lysosomal enzyme comprises an amino acid sequence which is at least 80% identical to the amino acid sequence of SEQ ID NO: 1 over its entire length.

### Claim 29 (depends on 14)

The method according to claim 14, wherein the lysosomal enzyme is transiently expressed in the target cell.

### Claim 30 (depends on 28)

The VLP according to claim 28, wherein the lysosomal enzyme comprises an amino acid sequence which is at least 90% identical to the amino acid sequence of SEQ ID NO: 1 over its entire length.

### Claim at position 31

31-32. (canceled)

### Claim 33 (depends on 1)

The method according to claim 1, wherein the subject is a human.

### Claim 34 (depends on 7)

The method according to claim 7, wherein the lysosomal enzyme comprises an amino acid sequence which is at least 90% identical to the amino acid sequence of SEQ ID NO: 1 over its entire length.

### Claim 35 (depends on 34)

The method according to claim 34, wherein the lysosomal enzyme comprises the amino acid sequence of SEQ ID NO: 1.

### Claim 36 (depends on 8)

The method according to claim 8, wherein the expression vector has a size of less than 6 kb.

### Claim 37 (depends on 10)

The method according to claim 10, wherein the nucleotide sequence encoding the lysosomal enzyme is at least 80% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length.

### Claim 38 (depends on 37)

The method according to claim 37, wherein the nucleotide sequence encoding the lysosomal enzyme is at least 90% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length.

### Claim 39 (depends on 38)

The method according to claim 38, wherein the nucleotide sequence encoding the lysosomal enzyme comprises the nucleotide sequence of SEQ ID NO: 2.

### Claim 40 (depends on 11)

The method according to claim 11, wherein said blood-brain barrier is a physiologically intact blood-brain barrier.

### Claim 41 (depends on 12)

The method according to claim 12, wherein the lysosomal enzyme or the expression vector enters an oligodendrocyte after crossing said blood-brain barrier.

### Claim 42 (depends on 1)

The method according to claim 1, wherein the VLP is administered intravenously.

### Claim 43 (depends on 15)

The method according to claim 15, wherein the lysosomal enzyme has a therapeutically effective enzyme activity for at least 20 days.

### Claim 44 (depends on 43)

The method according to claim 43, wherein the lysosomal enzyme has a therapeutically effective enzyme activity for at least 30 days.

### Claim 45 (depends on 17)

The method according to claim 17, wherein the VP1 protein comprises an amino acid sequence which is at least 90% identical to the amino acid sequence according to SEQ ID NO: 3 over its entire length.

### Claim 46 (depends on 20)

The expression vector according to claim 20, wherein the nucleotide sequence encoding the lysosomal enzyme comprises a nucleotide sequence which is at least 80% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length.

### Claim 47 (depends on 46)

The expression vector according to claim 46, wherein the nucleotide sequence encoding the lysosomal enzyme comprises a nucleotide sequence which is at least 90% identical to the nucleotide sequence of SEQ ID NO: 2 over its entire length.

### Claim 48 (depends on 47)

The expression vector according to claim 47, wherein the nucleotide sequence encoding the lysosomal enzyme comprises the nucleotide sequence of SEQ ID NO: 2.

### Claim 49 (depends on 19)

The expression vector according to claim 19, wherein the lysosomal enzyme is human arylsulfatase A.