# US 10,500,271 B2 · Compositions and methods for self-adjuvanting vaccines against microbes and tumors

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- Number: US 10,500,271 B2
- Document ID: US10500271B2
- Status: Granted
- Kind: B2
- Application number: 16/123,926
- Filed: 2018-09-06
- Published: 2018-12-27
- Granted: 2019-12-10
- Earliest claimed: 2010-03-23
- Assignees: The Regents of the University of California
- Applicants: The Regents of the University of California
- Inventors: Richard Syd Kornbluth; Geoffrey William Stone
- Web page: https://patentgrep.com/patent/US10500271B2

## CPC

- A61K 39/39: characterised by the immunostimulating additives, e.g. chemical adjuvants
- A61K 35/763: Herpes virus
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 31/12: Antivirals
- A61P 35/00: Antineoplastic agents
- C07K 14/005: from viruses
- C07K 14/70578: {NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 (NGF-receptor C07K14/71, TNF-receptor C07K14/7151)}
- C12N 15/86: Viral vectors
- A61K 2039/5256: {expressing foreign proteins}
- A61K 2039/55516: {Proteins; Peptides}
- C07K 2319/03: containing a transmembrane segment
- C12N 2710/16222: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2710/16632: Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
- C12N 2710/24143: viral genome or elements thereof as genetic vector
- C12N 2740/15011: Lentivirus, not HIV, e.g. FIV, SIV
- C12N 2740/16011: Human Immunodeficiency Virus, HIV
- 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)

## Abstract

The present invention is drawn to compositions and methods to enhance an immune response in order to prevent or treat infections or hyperproliferative diseases such as cancer. More particularly, the composition is an immunostimulatory intracellular signaling peptide fused directly or indirectly to a peptide that leads to multimerization into complexes of three or more units, where the intracellular signaling peptide must be present in a complex of three or more units in order to stimulate an immune response. Inserting this fusion construct into viruses like HIV-1 or introducing it into dendritic cells or tumor cells is predicted to lead to a positive therapeutic effect in humans, non-human mammals, birds, and fish.

## Claims

### Claim 1

An isolated cell, comprising:
(i) a first expression cassette comprising a nucleotide encoding a multimerizing domain from an N-terminal fragment of Epstein-Barr virus (EBV) Latent Membrane Protein 1 (LMP1), operatively joined to a nucleotide encoding a cytoplasmic signaling domain of LMP1, wherein the nucleotides do not contain introns; and
(ii) a second expression cassette for expressing an antigen, wherein the antigen is not an EBV protein.

### Claim 2 (depends on 1)

The isolated cell of claim 1, wherein the multimerizing domain comprises at least four transmembrane regions.

### Claim 3 (depends on 1)

The isolated cell of claim 1, wherein the antigen is endogenous to the cell.

### Claim 4 (depends on 1)

The isolated cell of claim 1, wherein the antigen is not normally expressed by the cell.

### Claim 5 (depends on 1)

The isolated cell of claim 1, wherein the antigen comprises a tumor antigen, viral antigen, or microbial antigen.

### Claim 6 (depends on 1)

The isolated cell of claim 1, wherein the cell is a microorganism.

### Claim 7 (depends on 6)

The isolated cell of claim 6, wherein the microorganism is a Listeria species or a Salmonella species.

### Claim 8 (depends on 1)

The isolated cell of claim 1, wherein the cell is a tumor cell.

### Claim 9

A virus, comprising:
(i) a first expression cassette comprising a multimerizing domain from the N-terminal fragment of Epstein-Barr virus (EBV) Latent Membrane Protein 1 (LMP1), operatively joined to a cytoplasmic signaling domain of LMP1, wherein the nucleotide encoding LMP1 does not contain introns; and
(ii) a second expression cassette for expressing an antigen, wherein the antigen is not an EBV protein.

### Claim 10 (depends on 9)

The virus of claim 9, wherein the multimerizing domain comprises at least four transmembrane regions.

### Claim 11 (depends on 9)

The virus of claim 9, wherein the antigen comprises a tumor antigen, viral antigen, or microbial antigen.

### Claim 12 (depends on 9)

The virus of claim 9, wherein the first and second expression cassettes are contained within the virus, wherein the virus is selected from the group consisting of human immunodeficiency virus-1 (HIV-1), simian immunodeficiency virus (SIV), influenza virus, parainfluenza virus, Hepatitis A virus, Hepatitis B virus, adenovirus, adeno-associated virus, Simian virus 40 (SV40), Modified Vaccinia Ankara (MVA), an arenavirus, a bunyavirus, a flavivirus, a herpesvirus, measles virus, a rhabdovirus, and combinations thereof.

### Claim 13 (depends on 12)

The virus of claim 12, wherein the herpesvirus is cytomegalovirus (CMV).

### Claim 14 (depends on 9)

The virus of claim 9, wherein the virus is an oncolytic virus.

### Claim 15

A herpes simplex virus 1 (HSV1) strain, lacking a functional ICP34.5-encoding gene, wherein the virus is derived from HSV1 strain JS1 as deposited at the European Collection of Cell Cultures (ECACC) under accession number 01010209; and wherein the virus comprises an expression cassette comprising a nucleotide sequence encoding a multimerizing domain from the N-terminal fragment of EBV LMP1, operatively joined to a nucleotide sequence encoding a cytoplasmic signaling domain of LMP1.

### Claim 16 (depends on 15)

The virus of claim 15, wherein the virus comprises a gene encoding an immunostimulatory protein, wherein the virus lacks a functional ICP34.5 encoding gene and a functional ICP47 encoding gene, and wherein the virus is replication competent in tumor cells.

### Claim 17 (depends on 15)

The virus of claim 15, wherein the nucleotide sequences do not contain intron sequences within the LMP1 open reading frames (ORFs).

### Claim 18 (depends on 15)

A method of treating cancer in a patient in need thereof, the method comprising administering to the patient an effective amount of the virus of claim 15.

### Claim 19 (depends on 12)

The virus of claim 12, wherein the flavivirus is selected from the group consisting of Hepatitis C virus, dengue virus, West Nile virus, Japanese Encephalitis virus, Venezuelan equine encephalitis virus, Eastern equine encephalitis virus and Western equine encephalitis.

### Claim 20 (depends on 12)

The virus of claim 12, wherein the rhabdovirus is vesicular stomatitis virus (VSV).