LIPID-BASED FORMULATIONS FOR THE DELIVERY OF RNA

Abstract

The invention provides a composition which is suitable for the delivery of RNA, which composition comprises (i) particles contained in a liquid phase, wherein the particles comprise RNA and a lipid composition, and (ii) 1,2 propanediol. The lipid composition comprises (i-a) a cholesterol derivative, (i-b) a phosphoglyceride, and (i-c) a pegylated phosphoglyceride. Further provided are a method for preparing the composition, and a solid composition which is obtainable by freezing the composition wherein particles are contained in a liquid phase.

Claims

1. A composition comprising (i) particles contained in a liquid phase, wherein the particles comprise RNA and a lipid composition, and wherein the lipid composition comprises: (i-a) a cholesterol derivative of formula (I) or a salt thereof: ##STR00012## wherein n is 0 or 1, preferably 0, R.sup.1 is a group —(CH.sub.2).sub.q—NH.sub.2 or a group —(CH.sub.2).sub.r—NH—(CH.sub.2).sub.s—NH.sub.2, wherein q, r and s are independently an integer of 2 to 6, R.sup.2 is a group —(CH.sub.2).sub.t—NH.sub.2 or a group —(CH.sub.2).sub.u—NH—(CH.sub.2).sub.w—NH.sub.2, wherein t, u and w are independently an integer of 2 to 6, R.sup.3 is a linear alkanediyl group having 1 to 4 carbon atoms; (i-b) a phosphoglyceride of formula (II) or a salt thereof: ##STR00013## wherein R.sup.4 is a linear alkyl group having 10 to 24 carbon atoms or a linear alkenyl group having 1 to 3 double bonds and 10 to 24 carbon atoms; R.sup.5 is a linear alkyl group having 10 to 24 carbon atoms or a linear alkenyl group having 1 to 3 double bonds and 10 to 24 carbon atoms; and (i-c) a pegylated phosphoglyceride of formula (III) or a salt thereof: ##STR00014## wherein p is an integer of 5 to 200, preferably 10 to 170 and most preferably 10 to 140 R.sup.6 is a linear alkyl group having 10 to 20 carbon atoms or a linear alkenyl group having 1 to 3 double bonds and 10 to 20 carbon atoms; R.sup.7 is a linear alkyl group having 10 to 20 carbon atoms or a linear alkenyl group having 1 to 3 double bonds and 10 to 20 carbon atoms; and (ii) 1,2-propanediol.

2. The composition according to claim 1, wherein the RNA is mRNA.

3. The composition according to claim 1, wherein the cholesterol derivative of formula (I) is GL67.

4. The composition according to claim 1, wherein the phosphoglyceride of formula (II) is 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine.

5. The composition according to claim 1, wherein the pegylated phosphoglyceride of formula (III) is 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-PEG5000.

6. The composition according to claim 1, wherein the molar ratio of the components (i-a):(i-b):(i-c) in the lipid composition is 1:(0.5 to 5):(0.01 to 1).

7. The composition according to claim 1, wherein the particles have average particle diameter in the range of 1 to 5000 nm.

8. The composition according to claim 1, wherein the N/P ratio of the number of nitrogen atoms N derived from the cholesterol derivative of formula (I) to the number of phosphate groups P in the RNA is in the range of 1 to 100.

9. The composition according to claim 1, wherein the 1,2-propanediol is contained at a concentration of 0.1 to 50% w/v, based on the total volume of the composition.

10. The composition according to claim 1, wherein the liquid phase in which the particles are contained comprises water.

11. A process for the preparation of a composition in accordance with claim 1, said process comprising the steps of: a) dissolving and mixing the components of the lipid composition in an organic solvent, followed by the lyophilization of the lipid composition; b) rehydrating the lyophilized lipid composition via addition of water; c) combining the rehydrated lipid composition with an aqueous solution of the RNA to allow particles comprising RNA and the lipid composition to be formed which are contained in a liquid phase; and d) adding 1,2-propanediol.

12. A solid composition comprising (i) particles comprising RNA and a lipid composition, and (ii) 1,2-propanediol, which solid composition is obtainable by freezing the composition according to claim 1.

13. A method of treating or preventing a disease via an RNA-based therapy using the composition of claim 1.

14. The method in accordance with claim 13 wherein the disease to be treated or prevented is a lung disease.

15. The method in accordance with claim 13, wherein the treatment or prevention involves the administration of the composition to or via the respiratory tract, preferably via pulmonary administration or nasal administration.

Description

[0324] Figures:

[0325] FIG. 1 shows the tdTomato level after transfection with GL67-complexes in different excipient solutions (water (0% propylene glycol) or 5% propylene glycol) at N/P 3 and 5.

[0326] FIG. 2 shows the luciferase activity in the nose of Balb/c mice 5h after treatment with GL67 complexes in water or in a 5% propylene glycol solution.

[0327] FIG. 3 shows the particle size before freezing and after thawing in the presence or absence of 1,2-propanediol.