# US 9,750,799 B2 · Broad spectrum influenza A neutralizing vaccines and D-peptidic compounds, and methods for making and using the same

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- Number: US 9,750,799 B2
- Document ID: US9750799B2
- Status: Granted
- Kind: B2
- Application number: 15/056,743
- Filed: 2016-02-29
- Published: 2016-08-04
- Granted: 2017-09-05
- Earliest claimed: 2011-12-02
- Assignees: Reflexion Pharmaceuticals, Inc.; The Governing Council of the University of Toronto
- Applicants: Reflexion Pharmaceuticals, Inc.; The Governing Council of the University of Toronto
- Inventors: Dana Ault-Riche; Maruti Uppalapati
- Web page: https://patentgrep.com/patent/US9750799B2

## CPC

- A61K 39/145: Orthomyxoviridae, e.g. influenza virus
- A61P 31/12: Antivirals
- A61P 31/16: for influenza or rhinoviruses
- C07K 14/005: from viruses
- C07K 14/11: Orthomyxoviridae, e.g. influenza virus
- 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 15/1037: {Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display}
- G01N 33/56983: {Viruses}
- G01N 33/68: involving proteins, peptides or amino acids {(involving lipoproteins G01N33/92)}
- G01N 33/6845: {Methods of identifying protein-protein interactions in protein mixtures}
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C12N 2760/16122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2760/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- G01N 2333/11: Orthomyxoviridae, e.g. influenza virus
- 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
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

GB1 peptidic compounds that specifically bind to a hemagglutinin target protein, and libraries that include the same, as well as methods of making and using the same, are provided. Also provided are methods and compositions for making and using the compounds. Also provided are hemagglutinin mimics and fragments and methods of using the same, including methods of screening for GB1 peptidic compounds and methods of using conjugates the mimics as influenza A vaccines. Aspects of the invention include methods of screening libraries of L-peptidic compounds for specific binding to a D-peptidic hemagglutinin target protein. Once a L-peptidic compound has been identified that specifically binds to the D-peptidic hemagglutinin target protein, the D-enantiomer of the selected L-peptidic compound may be produced. In some embodiments, the D-enantiomer of the selected L-peptidic compound binds to, and in some instances, neutralizes influenza virus particles.

## Claims

### Claim 1

A conjugate comprising:
an L-peptidic coiled coil hemagglutinin mimic comprising two covalently linked strands that each independently comprise a peptidic sequence having one of SEQ ID NOs:87-102; and
a carrier protein conjugated to the L-peptidic coiled coil hemagglutinin mimic.

### Claim 2 (depends on 1)

A method of inducing an antibody response in a subject in need thereof, the method comprising administering a conjugate to the subject in an amount sufficient to induce an antibody response in the subject, wherein the conjugate comprises the L-peptidic coiled coil hemagglutinin mimic of claim 1.

### Claim 3 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic is described by one of formulae (X) and (XI):
wherein Z1 and Z2 each comprise a peptidic hemagglutinin epitope displayed on a coiled coil scaffold;
T1 and T2 are optional tethers; and
L is a linker.

### Claim 4 (depends on 3)

The conjugate of claim 3, wherein Z1 and Z2 each independently comprise a peptidic sequence of one of SEQ ID NOs:87-102 wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 5 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence QKSaQdIDGaTSdVNSaIEd (SEQ ID NO:87), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 6 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence KESaQKdIDGaTNdVNSaIEd (SEQ ID NO:88), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 7 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence LKSaQAdIDQaNGdLNRaIEd (SEQ ID NO:89), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 8 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence KESaQKdIDGaTNdVNSaIDd (SEQ ID NO:91), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 9 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence RESaQKdVDGaTNdVNSaIDd (SEQ ID NO:92), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 10 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence YKSaQSdIDQaTGdLNRaIEd (SEQ ID NO:93), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 11 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence QKSaQEdIDKaTNdVNNaVDd (SEQ ID NO:94), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 12 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence KGSaQKdIDKaTSdVNNaIDd (SEQ ID NO:95), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 13 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence YKSaQAdIDQaTGdLNRaIEd (SEQ ID NO:96), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 14 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence KESaQKdIDQaTSdVNNaVDd (SEQ ID NO:97), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 15 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence RDSaQRdIDNaQNdLNNaIDd (SEQ ID NO:98), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 16 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence KESaQKdIDQaTTdINNaIDd (SEQ ID NO:99), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 17 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence LKSaQAdIDQaNGdLNRaIEd (SEQ ID NO:100), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 18 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence YKSaQAdIDQaTGdLNRaIEd (SEQ ID NO:101), wherein each “a” and each “d” are independently selected from Ile and Leu.

### Claim 19 (depends on 1)

The conjugate of claim 1, wherein the L-peptidic coiled coil hemagglutinin mimic comprises a peptidic sequence KASaQKdIDEaTTdINNaIEd (SEQ ID NO:102), wherein each “a” and each “d” are independently selected from Ile and Leu.