Cationic oil-in-water emulsions

Patent metadata

Field	Value
Publication number	US-9636410-B2
Application number	US-201214130886-A
Country	US
Kind code	B2
Filing date	Jul 6, 2012
Priority date	Jul 6, 2011
Publication date	May 2, 2017
Grant date	May 2, 2017

Abstract

This invention generally relates to cationic oil-in-water emulsions that contain high concentrations of cationic lipids and have a defined oil:lipid ratio. The cationic lipid can interact with the negatively charged molecule thereby anchoring the molecule to the emulsion particles. The cationic emulsions described herein are useful for delivering negatively charged molecules, such as nucleic acid molecules to cells, and for formulating nucleic acid-based vaccines.

First claim

The invention claimed is:

1. A stable oil-in-water emulsion comprising particles that are dispersed in an aqueous continuous phase, wherein the average diameter of said particles is from about 80 nm to 150 nm, wherein the average diameter does not change by more than 10% when the emulsion is stored at 4° C. for one month; and wherein the emulsion comprises an oil and a cationic lipid, wherein the head group of the cationic lipid comprises a quaternary amine, and
wherein:
(i) the ratio of oil:lipid (mole:mole) is at least about 8:1 (mole:mole);
(ii) the concentration of cationic lipid in said emulsion is at least about 2.5 mM; and
(iii) the cationic lipid is not DC-Cholesterol.

2. The oil-in-water emulsion of claim 1 , wherein the average diameter of said particles is from about 80 nm to about 130 nm.

3. The oil-in-water emulsion of claim 1 , wherein the ratio of oil:lipid (mole:mole) is from about 10:1 (mole:mole) to about 43:1 (mole:mole).

4. The oil-in-water emulsion of claim 1 , wherein said oil-in-water emulsion comprises from about 0.2% to about 8% (w/v) oil.

5. The oil-in-water emulsion of claim 4 , wherein said oil is squalene or squalane.

6. The oil-in-water emulsion of claim 1 , wherein the oil-in-water emulsion further comprises from about 0.01% to about 2.5% (v/v) surfactant.

7. The oil-in-water emulsion of claim 6 , wherein said surfactant is SPAN85 (SorbitanTrioleate), Tween 80 (polysorbate 80), or a combination thereof.

8. The oil-in-water emulsion of claim 7 , wherein the oil-in-water emulsion comprises about 0.5% (v/v) Tween 80 and about 0.5% (v/v) SPAN85.

9. The oil-in-water emulsion of claim 1 , wherein the cationic lipid is selected from the group consisting of: 1,2-dioleoyloxy-3-(trimethylammonio)propane (DOTAP), 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine (DOEPC), N,N-dioleoyl-N,N-dimethylammonium chloride (DODAC), and N-[1-(2, 3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA).

10. The oil-in-water emulsion of claim 9 , wherein the cationic lipid is DOTAP and the concentration of DOTAP in said emulsion is from about 2.58 mM (1.8 mg/mL) to about 7.16 mM (5 mg/mL).

11. A method for preparing the oil-in-water emulsion of claim 1 , comprising: (a) directly dissolving the cationic lipid in the oil to form an oil phase; (b) providing an aqueous phase of the emulsion; and (c) dispersing the oil phase in the aqueous phase by homogenization.

12. The method of claim 11 , wherein step (a) further comprises heating the oil to a temperature between about 30° C. to about 65° C.

13. The oil-in-water emulsion of claim 1 further comprising an antioxidant.

Assignees

• Brito Luis
• Chan Michelle
• Geall Andrew
• O'Hagan Derek
• Singh Manmohan
• Glaxosmithkline Biologicals Sa

Inventors

• Brito Luis
• Chan Michelle
• Geall Andrew
• O'Hagan Derek
• Singh Manmohan

Classifications

• C12N2770/36143

  viral genome or elements thereof as genetic vector · CPC title

• A61K47/24

  containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids · CPC title

• C12N2710/16171

  Demonstrated in vivo effect · CPC title

• A61K47/34

  Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers (A61K47/10 takes precedence) · CPC title

• A61K39/39 Primary

  characterised by the immunostimulating additives, e.g. chemical adjuvants · CPC title