Delivery of self-replicating RNA using biodegradable polymer particles
Particle compositions comprising adsorbed RNA replicons as well as methods of making and using the same are described.
1. An immunogenic composition comprising:
(a) positively charged nanoparticles that comprise a biodegradable polymer and greater than 1% (w/w) of a cationic surfactant, wherein:
(i) the biodegradable polymer is a poly(α-hydroxy acid), and
(ii) the nanoparticles have Z average mean particle size value that is between 100 and 500 nanometers, and a zeta potential greater than +50 mV;
(b) an RNA replicon comprising at least one polynucleotide encoding at least one antigen adsorbed to said positively charged nanoparticles; and
(c) a non-ionic surfactant.
2. The immunogenic composition of claim 1 , wherein the biodegradable polymer is a poly(lactide-co-glycolide).
3. The immunogenic composition of claim 1 , wherein the biodegradable polymer is a poly(lactide-co-glycolide) having a lactide:glycolide molar ratio ranging from 40:60 to 60:40.
4. The immunogenic composition of claim 1 , wherein the cationic surfactant is selected from (1,2-dioleoyloxypropyl)-N,N,N-trimethylammonium salt (DOTAP), dimethyldioctadecylammonium salt (DDA), and 3-beta-[N—(N′,N′-dimethylaminoethane)carbamoyl]cholesterol (DC-Chol).
5. The immunogenic composition of claim 1 , wherein the cationic surfactant comprises an ammonium group and a saturated or unsaturated hydrocarbon chain having between 12 to 20 carbon atoms.
6. The composition of claim 1 , wherein said non-ionic surfactant is poly(vinyl alcohol).
7. The immunogenic composition of claim 1 , wherein said composition further comprises at least one additional component selected from polyols, carbohydrates and combinations thereof.
8. The immunogenic composition of claim 7 , wherein said at least one additional component comprises an alditol and a saccharide.
9. The immunogenic composition of claim 1 , wherein said RNA replicon is an alphavirus replicon.
10. The immunogenic composition of claim 9 , wherein the alphavirus replicon is derived from an alphavirus selected from the group consisting of: Sindbis (SIN), Venezuelan equine encephalitis (VEE), Semliki Forest virus (SFV) and combinations thereof.
11. The immunogenic composition of claim 1 , wherein the at least one antigen is selected from a viral antigen, a bacterial antigen and a tumor antigen.
12. The immunogenic composition of claim 1 , wherein the at least one antigen is selected from an influenza virus, a respiratory syncytial virus (RSV), a parainfluenza virus (PIV), hepatitis B virus (HBV), a hepatitis C virus (HCV), a human immunodeficiency virus (HIV), a herpes simplex virus (HSV), and a human papilloma virus (HPV), yellow fever, pandemic flu, tuberculosis, dengue, norovirus, measles, rhinovirus, west nile virus, polio, hepatitis A and cytomegalo virus (CMV).
13. The immunogenic composition of claim 1 , further comprising an immunological adjuvant.
14. The immunogenic composition of claim 13 , wherein the immunological adjuvant is selected from small molecule immune potentiators, toll like receptor ligands, and immunostimulatory oligonulcleotides.
15. The immunogenic composition of claim 13 , wherein the immunological adjuvant is adsorbed or entrapped within nanoparticles that may be the same or different from the positively charged nanoparticles comprising the adsorbed RNA replicon.
16. The immunogenic composition of claim 1 , wherein the immunogenic composition is an injectable composition.
17. The immunogenic composition of claim 1 , wherein the immunogenic composition is lyophilized.
18. An immunogenic composition comprising:
(a) positively charged nanoparticles that comprise a biodegradable polymer and about 4% (w/w) of a cationic surfactant selected from (1,2-dioleoyloxypropyl)-N,N,N-trimethylammonium salt (DOTAP), dimethyldioctadecylammonium salt (DDA), and 3-beta-[N—(N′,N′-dimethylaminoethane)carbamoyl]cholesterol (DC-Chol) wherein:
(i) the biodegradable polymer is a poly(lactide-co-glycolide), and
(ii) the nanoparticles have Z average mean particle size value that is between 100 and 250 nanometers, and a zeta potential greater than +50 mV, and
(b) an RNA replicon comprising at least one polynucleotide encoding at least one antigen adsorbed to said positively charged nanoparticles, wherein ratio of the number of moles of cationic nitrogen in said cationic surfactant to the number of moles of anionic phosphate in said RNA replicon (N:P ratio) is about 4:1, or more.
19. A method of stimulating an immune response in a vertebrate host animal comprising administering to the animal the immunogenic composition of claim 1 .
20. A method of forming the immunogenic composition of claim 1 , comprising: (a) combining (i) a first liquid that comprises said biodegradable polymer and said cationic surfactant dissolved in an organic solvent with (ii) a second liquid that comprises water, whereupon a first suspension of nanoparticles comprising said biodegradable polymer and said cationic surfactant is formed, (b) adsorbing said RNA replicon to said nanoparticles in said first suspension to form a second suspension, (c) adding at least one additional component comprising a non-ionic surfactant to said second suspension to form a third suspension, and (d) lyophilizing said third suspension.
21. The method of claim 20 , wherein said at least one additional component is selected from polyols, carbohydrates and combinations thereof.
22. The composition of claim 18 , wherein the nanoparticles have Z average mean particle size value that is between about 200 and about 250 nanometers with a polydispersity index of less than about 0.2.
23. The composition of claim 22 , wherein the nanoparticles are formed by solvent extraction, wherein the poly(lactide-co-glycolide) and cationic surfactant are dissolved in acetone solvent.
24. The composition of claim 23 , wherein the cationic surfactant is (1,2-dioleoyloxypropyl)-N,N,N-trimethylammonium salt (DOTAP).
25. The composition of claim 24 , wherein the composition further comprises about 0.5 to about 20% (w/w) non-ionic surfactant.
26. The composition of claim 1 , wherein:
(a) the nanoparticles have Z average mean particle size value that is between about 200 and about 250 nanometers with a polydispersity index of less than about 0.2,
(b) the cationic surfactant is (1,2-dioleoyloxypropyl)-N,N,N-trimethylammonium salt (DOTAP), which is present at about 4% (w/w), and
(c) the nanoparticles are formed by solvent extraction, wherein the poly(α-hydroxy acid) and cationic surfactant are dissolved in acetone solvent.