# US 10,214,565 B2 · Compositions and methods for treating, including preventing, parvovirus infections and related diseases

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- Number: US 10,214,565 B2
- Document ID: US10214565B2
- Status: Granted
- Kind: B2
- Application number: 15/124,699
- Filed: 2015-03-10
- Published: 2017-01-19
- Granted: 2019-02-26
- Earliest claimed: 2014-03-10
- Assignees: University of Louisville Research Foundation, Inc.
- Applicants: University of Louisville Research Foundation, Inc.
- Inventors: Shin-je Ghim; A. Bennett Jenson; John O. Trent
- Web page: https://patentgrep.com/patent/US10214565B2

## CPC

- C07K 14/005: from viruses
- A61K 39/12: Viral antigens
- A61K 39/23: Parvoviridae, e.g. feline panleukopenia virus
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 7/06: Antianaemics
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 31/20: for DNA viruses
- A61P 37/02: Immunomodulators
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- C12N 15/85: for animal cells
- C12P 21/02: having a known sequence of two or more amino acids, e.g. glutathione
- A61K 2039/5258: {Virus-like particles}
- A61K 2039/55511: {Organic adjuvants}
- A61K 2039/55566: {Emulsions, e.g. Freund's adjuvant, MF59}
- A61K 2039/575: {humoral response}
- C12N 2710/14043: viral genome or elements thereof as genetic vectore
- C12N 2750/14222: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2750/14223: Virus like particles [VLP]
- C12N 2750/14234: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Some embodiments of the invention include inventive polypeptides (e.g., mutant VP2 proteins) and virus-like particles made from the inventive polypeptides. Other embodiments of the invention include compositions for treating (e.g., preventing) parvovirus (e.g., erythrovirus or parvovirus B19) infection and other diseases. Further embodiments include methods for administering compositions to an animal. Other embodiments include treating (e.g., preventing) parvovirus (e.g., erythrovirus or parvovirus B19) infection and other diseases. Still other embodiments include nucleic acid sequences that encode the inventive polypeptides. Additional embodiments of the invention are also discussed.

## Claims

### Claim 1

A polypeptide comprising a VP2 polypeptide with at least one amino acid modification relative to wild type VP2
wherein
the wild type VP2 has the amino acid sequence of SEQ ID NO: 1 and
the at least one amino acid modification comprises a substitution at Y401, a substitution at Q399, a substitution at Q400, a substitution at Q404, a substitution at Q368, a substitution at Q369, a substitution at Y392, Y401F, Y401W, Y401A, Q368A, Q369A, Q368N, Q369N, Q399N, Q400N, Q404T, Y392A, Y392F, Q404N, Y401P, T402A, D403A, Q404A, or combinations thereof.

### Claim 2 (depends on 1)

The polypeptide of claim 1, wherein the at least one amino acid modification further comprises an insertion, a deletion, or a substitution.

### Claim 3 (depends on 1)

The polypeptide of claim 1, wherein the polypeptide sequence has at least about 90% identity to SEQ ID NO: 1.

### Claim 4 (depends on 1)

The polypeptide of claim 1, wherein the at least one amino acid modification further comprises an amino acid modification in the recess of the three-fold axis or less than about 50 Å from the recess of the three-fold axis.

### Claim 5 (depends on 1)

The polypeptide of claim 1, wherein the at least one amino acid modification further comprises a substitution in loop 3, a substitution in loop 4, a substitution upstream of loop 3, a substitution upstream of loop 4, or combinations thereof.

### Claim 6 (depends on 1)

The polypeptide of claim 1, wherein the at least one amino acid modification comprises Q399N, Q400N, Y401F, Q404T, or combinations thereof.

### Claim 7 (depends on 1)

The polypeptide of claim 1, wherein the polypeptide is Construct F.

### Claim 8 (depends on 1)

The polypeptide of claim 1, wherein the polypeptide is selected from the group consisting of construct A, construct B, construct C, construct D, construct E, construct F, construct G, construct H, construct I, construct J, construct K, construct L, construct M, construct N, construct O, construct P, construct Q, construct R, construct S, construct T, construct U, construct V, construct W, and construct X.

### Claim 9 (depends on 1)

The polypeptide of claim 1, wherein the polypeptide sequence has at least about 95% identity to SEQ ID NO: 1.

### Claim 10 (depends on 1)

An mVLP comprising the polypeptide of claim 1, wherein
the at least one amino acid modification comprises (a) Y401F and (b) Q399N or Q404T, and
the polypeptide is not construct J.

### Claim 11 (depends on 10)

The mVLP of claim 10, wherein the mVLP has reduced binding to P antigen compared to wtVLP, the mVLP has no detectable binding to P antigen, the mVLP has reduced hemagglutination of red blood cells compared to wtVLP, or the mVLP has no detectable hemagglutination.

### Claim 12 (depends on 10)

The mVLP of claim 10, wherein the mVLP has one or more neutralizing epitopes.

### Claim 13 (depends on 10)

The mVLP of claim 10, wherein the mVLP induces the production of antibodies in an animal, where the antibodies produced are capable of reducing or inhibiting hemagglutination by wtVLP.

### Claim 14 (depends on 10)

A composition comprising the mVLP of claim 10.

### Claim 15 (depends on 10)

An immunogenic pharmaceutical composition comprising the mVLP of claim 10.

### Claim 16 (depends on 15)

The immunogenic composition of claim 15, wherein the amount of mVLP is from about 0.0001% (by weight total composition) to about 50%.

### Claim 17 (depends on 15)

The immunogenic composition of claim 15, further comprising an adjuvant or a carrier.

### Claim 18 (depends on 15)

The immunogenic composition of claim 15, wherein the immunogenic composition does not comprise an adjuvant.

### Claim 19 (depends on 10)

A method for providing an animal with an mVLP comprising one or more administrations of one or more compositions comprising the mVLP of claim 10, wherein the compositions may be the same or different if there is more than one administration.

### Claim 20 (depends on 19)

The method of claim 19, wherein at least one of the one or more compositions does not comprise an adjuvant.

### Claim 21 (depends on 19)

The method of claim 19, wherein at least one of the one or more compositions further comprises a carrier or an adjuvant.

### Claim 22 (depends on 19)

The method of claim 19, wherein if there is more than one administration at least one composition used for at least one administration is different from the composition of at least one other administration.

### Claim 23 (depends on 10)

A method for inducing an immune response in an animal comprising one or more administrations of one or more compositions comprising the mVLP of claim 10, wherein the compositions may be the same or different if there is more than one administration.

### Claim 24 (depends on 23)

The method of claim 23, wherein at least one of the one or more compositions does not comprise an adjuvant.

### Claim 25 (depends on 23)

The method of claim 23, wherein at least one of the one or more compositions further comprises a carrier or an adjuvant.

### Claim 26 (depends on 23)

The method of claim 23, wherein if there is more than one administration at least one composition used for at least one administration is different from the composition of at least one other administration.

### Claim 27 (depends on 1)

A method for producing a polypeptide of claim 1 comprising,
culturing a host cell transfected with a vector comprising a nucleic acid sequence encoding the polypeptide, to provide expression of the polypeptide; and
recovering the polypeptide.

### Claim 28 (depends on 10)

A method for producing the mVLP of claim 10 comprising,
culturing a host cell transfected with a vector comprising a nucleic acid sequence encoding the polypeptide, to provide expression of the polypeptide; and
recovering the mVLP.

### Claim 29 (depends on 1)

An isolated nucleic acid molecule encoding the polypeptide of claim 1.

### Claim 30 (depends on 29)

The nucleic acid molecule of claim 29, wherein the nucleic acid molecule is included in a vector or plasmid.

### Claim 31 (depends on 29)

A vector comprising the nucleic acid molecule of claim 29.

### Claim 32 (depends on 10)

The mVLP of claim 10, wherein the at least one amino acid modification comprises Y401F and Q399N.

### Claim 33 (depends on 10)

The mVLP of claim 10, wherein the at least one amino acid modification comprises Y401F and Q404T.

### Claim 34 (depends on 10)

The mVLP of claim 10, wherein the at least one amino acid modification does not comprises Q404N.

### Claim 35 (depends on 10)

The mVLP of claim 10, wherein the at least one amino acid modification comprises Y401F, Q399N, and Q404T.

### Claim 36 (depends on 10)

The mVLP of claim 10, wherein the polypeptide is Construct F.