# US 10,307,374 B2 · Oil-in-water emulsions that contain nucleic acids

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- Number: US 10,307,374 B2
- Document ID: US10307374B2
- Status: Granted
- Kind: B2
- Application number: 15/495,066
- Filed: 2017-04-24
- Published: 2017-08-10
- Granted: 2019-06-04
- Earliest claimed: 2011-07-06
- Assignees: Glaxosmithkline Biologicals SA
- Applicants: Glaxosmithkline Biologicals SA
- Inventors: Luis Brito; Michelle C. Archer; Andrew Geall; Derek O'Hagan; Manmohan Singh
- Web page: https://patentgrep.com/patent/US10307374B2

## CPC

- A61K 9/107: Emulsions {; Emulsion preconcentrates; Micelles (composition of emulsions A61K47/00)}
- A61K 31/713: Double-stranded nucleic acids or oligonucleotides
- A61K 39/12: Viral antigens
- A61K 39/155: Paramyxoviridae, e.g. parainfluenza virus
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 25/16: Anti-Parkinson drugs
- A61P 25/28: for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
- A61P 31/04: Antibacterial agents
- A61P 31/10: Antimycotics
- A61P 31/12: Antivirals
- A61P 31/18: for HIV
- A61P 33/02: Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
- A61P 35/00: Antineoplastic agents
- A61P 37/02: Immunomodulators
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- A61P 37/08: Antiallergic agents (antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14)
- A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)}
- A61K 2039/53: {DNA (RNA) vaccination}
- A61K 2039/55566: {Emulsions, e.g. Freund's adjuvant, MF59}
- C12N 2760/18534: 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
- 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)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

This invention generally relates to cationic oil-in-water emulsions that can be used to deliver nucleic acid molecules, such as an RNA molecule. The emulsion particles comprise an oil core and a cationic lipid. The emulsion particles have an average diameter of about 80 nm to about 180 nm, and the emulsion have an N/P ratio of at least 1.1:1.

## Claims

### Claim 1

A method for generating an immune response in a subject, comprising administering to the subject an effective amount of an immunogenic cationic oil-in-water emulsion comprising emulsion particles that contain an oil core and a cationic lipid, and a nucleic acid molecule that is complexed to the emulsion particles, wherein:
a) the nucleic acid is RNA that encodes a polypeptide antigen,
b) the average diameter of the emulsion particles is from about 80 nm to about 180 nm, and
c) the N/P of the emulsion is at least 4:1;
with the proviso that the nucleic acid molecule does not encode secreted alkaline phosphatase, and the further proviso that the nucleic acid molecule is not the RNA encoded by the plasmid A317, the sequence of which is given by SEQ ID NO:1;
whereby the immune response is generated in the subject.

### Claim 2 (depends on 1)

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

### Claim 3 (depends on 1)

The method of claim 1, wherein the immunogenic cationic oil-in-water emulsion is buffered, and has a pH from about 6.0 to about 8.0.

### Claim 4 (depends on 3)

The method of claim 3, wherein the immunogenic cationic oil-in-water emulsion comprises a buffer selected from the group consisting of a citrate buffer, a succinate buffer, an acetate buffer, and combinations thereof.

### Claim 5 (depends on 4)

The method of claim 4, wherein the buffer is a citrate buffer and the pH is about 6.5.

### Claim 6 (depends on 1)

The method of claim 1, further comprising an inorganic salt.

### Claim 7 (depends on 6)

The method of claim 6, wherein the inorganic salt concentration is no greater than 30 mM.

### Claim 8 (depends on 1)

The method of claim 1, further comprising a nonionic tonicifying agent.

### Claim 9 (depends on 8)

The method of claim 8, wherein the nonionic tonicifying agent is a sugar, a sugar alcohol or combinations thereof.

### Claim 10 (depends on 9)

The method of claim 9, wherein the nonionic tonicifying agent is selected from the group consisting of sucrose, trehalose, sorbitol, dextrose and combinations thereof.

### Claim 11 (depends on 1)

The method of claim 1, further comprising a polymer in the aqueous phase.

### Claim 12 (depends on 11)

The method of claim 11, wherein the polymer is a poloxamer.

### Claim 13 (depends on 11)

The method of claim 11, wherein the polymer is Pluronic F127.

### Claim 14 (depends on 11)

The method of claim 11, wherein the emulsion contains about 0.05% to about 20% (w/v) polymer.

### Claim 15 (depends on 1)

The method of claim 1, wherein the average diameter of the emulsion particles is from about 80 nm to about 130 nm.

### Claim 16 (depends on 1)

The method of claim 1, wherein the N/P of the emulsion is from 4:1 to about 20:1.

### Claim 17 (depends on 1)

The method of claim 1, wherein the N/P of the emulsion is from 4:1 to about 15:1.

### Claim 18 (depends on 1)

The method of claim 1, wherein the cationic oil-in-water emulsion is isotonic with human blood.

### Claim 19 (depends on 1)

The method of claim 1, wherein the oil core comprises an oil that is selected from the group consisting of: Castor oil, Coconut oil, Corn oil, Cottonseed oil, Evening primrose oil, Fish oil, Jojoba oil, Lard oil, Linseed oil, Olive oil, Peanut oil, Safflower oil, Sesame oil, Soybean oil, Squalene, Squalane, Sunflower oil, Wheatgerm oil, and combinations thereof.

### Claim 20 (depends on 19)

The method of claim 19, wherein the oil is Squalene.

### Claim 21 (depends on 1)

The method of claim 1, wherein the cationic lipid is selected from the group consisting of: 1,2-dioleoyloxy-3-(trimethylammonio)propane (DOTAP), 3β-[N—(N′,N′-Dimethylaminoethane)-carbamoyl]Cholesterol (DC Cholesterol), dimethyldioctadecylammonium (DDA), 1,2-Dimyristoyl-3-TrimethylAmmoniumPropane (DMTAP), dipalmitoyl(C16:0)trimethyl ammonium propane (DPTAP), distearoyltrimethylammonium propane (DSTAP), N-[1-(2, 3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA), N,N-dioleoyl-N,N-dimethylammonium chloride (DODAC), 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine (DOEPC), 1,2-dioleoyl-3-dimethylammonium-propane (DODAP), 1,2-dilinoleyloxy-3-dimethylaminopropane (DLinDMA), Lipids E0001-E0118, and combinations thereof.

### Claim 22 (depends on 21)

The method of claim 21, wherein the cationic lipid is DOTAP.

### Claim 23 (depends on 1)

The method of claim 1, wherein the cationic lipid comprises a quaternary amine.

### Claim 24 (depends on 1)

The method of claim 1, wherein the particle further comprises a surfactant.

### Claim 25 (depends on 24)

The method of claim 24, wherein the surfactant is a nonionic surfactant.

### Claim 26 (depends on 25)

The method of claim 25, wherein the surfactant is Sorbitan Trioleate, polysorbate 80, or a combination thereof.

### Claim 27 (depends on 1)

The method of claim 1, wherein the emulsion further comprises an antioxidant.