# US 11,026,964 B2 · Delivery of RNA to different cell types

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- Number: US 11,026,964 B2
- Document ID: US11026964B2
- Status: Granted
- Kind: B2
- Application number: 15/691,042
- Filed: 2017-08-30
- Published: 2018-02-22
- Granted: 2021-06-08
- Earliest claimed: 2010-07-06
- Assignees: Glaxosmithkline Biologicals SA
- Applicants: Glaxosmithkline Biologicals, SA
- Inventors: Andrew Geall; Katrin Ramsauer; Gillis Otten; Christian Mandl
- Web page: https://patentgrep.com/patent/US11026964B2

## CPC

- A61K 31/7088: Compounds having three or more nucleosides or nucleotides
- A61K 39/12: Viral antigens
- A61K 39/155: Paramyxoviridae, e.g. parainfluenza virus
- C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids}
- A61K 2039/5256: {expressing foreign proteins}
- A61K 2039/53: {DNA (RNA) vaccination}
- A61K 2039/55505: {Inorganic adjuvants}
- A61K 2039/55555: {Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers}
- A61K 2039/55566: {Emulsions, e.g. Freund's adjuvant, MF59}
- C12N 2320/32: Special delivery means, e.g. tissue-specific
- C12N 2760/18534: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2770/36143: viral genome or elements thereof as genetic vector
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

RNA encoding an immunogen is co-delivered to non-immune cells at the site of delivery and also to immune cells which infiltrate the site of delivery. The responses of these two cell types to the same delivered RNA lead to two different effects, which interact to produce a strong immune response against the immunogen. The non-immune cells translate the RNA and express the immunogen. Infiltrating immune cells respond to the RNA by expressing type I interferons and pro-inflammatory cytokines which produce a local adjuvant effect which acts on the immunogen-expressing non-immune cells to upregulate major histocompatibility complex expression, thereby increasing presentation of the translated protein to T cells. The effects on the immune and non-immune cells can be achieved by a single delivery of a single RNA e.g. by a single injection.

## Claims

### Claim 1

A method of administering to a delivery site in a vertebrate
a pharmaceutical composition, in an immunologically effective amount to raise an effective immune response to a pathogen in the vertebrate, comprising immunogen encoding RNA encapsulated within liposomes and immunogen encoding RNA on the surface of liposomes, wherein at least half of the RNA is liposome-encapsulated, and wherein the RNA has a size of about 5000-25000 nucleotides and is not virion-packaged, and wherein the liposomes have an aqueous core,
and then permitting immune cells to infiltrate the delivery site, such that the RNA separately enters both (a) non-immune cells at the delivery site and (b) the infiltrating immune cells, provided that the RNA includes no modified nucleotides other than a 5′ cap.

### Claim 2 (depends on 1)

The method of claim 1, wherein the delivery site is in skeletal muscle tissue.

### Claim 3 (depends on 1)

The method of claim 1, wherein the RNA is administered by injection.

### Claim 4 (depends on 3)

The method of claim 3, wherein injection is via a needle.

### Claim 5 (depends on 1)

The method of claim 1, wherein the non-immune cells comprise muscle cells and/or fibroblasts.

### Claim 6 (depends on 1)

The method of claim 1, wherein the RNA is translated by the non-immune cells.

### Claim 7 (depends on 1)

The method of claim 1, wherein the immune cells comprise macrophages, dendritic cells and/or monocytes.

### Claim 8 (depends on 1)

The method of claim 1, wherein the immune cells are TLR7-positive.

### Claim 9 (depends on 1)

The method of claim 1, wherein the RNA causes the immune cells to secrete type I interferons and/or pro-inflammatory cytokines.

### Claim 10 (depends on 1)

The method of claim 1, wherein the RNA is +-stranded and can be translated by the non-immune cells.

### Claim 11 (depends on 1)

The method of claim 1, wherein the RNA can bind to TLR7.

### Claim 12 (depends on 1)

The method of claim 1, wherein the RNA is a self-replicating RNA.

### Claim 13 (depends on 1)

The method of claim 1, wherein the RNA encodes an immunogen which can elicit an immune response against a bacterium, a virus, a fungus, an allergen, a tumour, or a parasite.

### Claim 14 (depends on 13)

The method of claim 13, wherein the immunogen can elicit an immune response in vivo against respiratory syncytial virus glycoprotein F.

### Claim 15 (depends on 1)

The method of claim 1, wherein the size of the RNA is about 8000-15000 nucleotides.

### Claim 16 (depends on 1)

The method of claim 1, wherein the size of the RNA is about 9000-12000 nucleotides.

### Claim 17 (depends on 1)

The method of claim 1, wherein a size of the liposome is between 50-220 nm.

### Claim 18 (depends on 1)

The method of claim 1, wherein an amount of RNA encapsulated within the liposome is between 0.01 μg and 10 μg.