Methods and compositions for liposomal formulation of antigens and uses thereof

Patent metadata

Field	Value
Publication number	US-9931419-B2
Application number	US-201615181028-A
Country	US
Kind code	B2
Filing date	Jun 13, 2016
Priority date	Mar 9, 2009
Publication date	Apr 3, 2018
Grant date	Apr 3, 2018

Abstract

The present invention relates to liposomal vaccine compositions, methods for the manufacture thereof, and methods for the use thereof to stimulate an immune response in an animal. These compositions comprise dimyristoylphosphatidylcholine (“DMPC”); either dimyristoylphosphatidylglycerol (“DMPG”) or dimyristoyltrimethylammonium propane (“DMTAP”) or both DMPC and DMTAP; and at least one sterol derivative providing a covalent anchor for one or more immunogenic polypeptide(s) or carbohydrate(s).

First claim

We claim:

1. A composition comprising:
a) an aqueous vehicle; and
b) liposomes comprising
(i) dimyristoylphosphatidylcholine (“DMPC”),
(ii) one or more lipids selected from the group consisting of dimyristoylphosphatidylglycerol (“DMPG”), and dimyristoyltrimethylammonium propane (“DMTAP”), and
(iii) at least one sterol derivative,

at relative percentages of 50%-98% (i):1%-25% (ii):1%-25% (iii); and
wherein said sterol derivative comprises a reactive group selected from the group consisting of imidoesters, N-hydroxysuccinimidyl (“NHS”) esters, maleimides, alkyl halides, aryl halides, α-haloacyls, pyridyl disulfides, carbodiimides, glyoxals, amines, hydrazides, and arylazides.

2. A composition according to claim 1 , wherein the said sterol derivative comprises a maleimide reactive group.

3. A composition according to claim 1 , wherein the sterol derivative comprises a covalent linkage between the sterol derivative and the reactive group, the covalent linkage having the structure St-R3-RG, wherein:
St is a ring atom of the sterol derivative;
R3 is C 0-18 straight or branched chain alkyl, or C 0-12 straight or branched chain alkyl-(alkylene oxide) n -C 0-12 straight or branched chain alkyl, wherein n is on average between 40 and 1000;
each said straight or branched chain alkyl comprises from 0-4 chain heteroatoms and one or more substituents independently selected from the group consisting of halogen, trihalomethyl, —C 1-6 alkoxy, —NO 2 , —NH 2 , —OH, —CH 2 OH, —CONH 2 , and —C(O)(OR4) where R4 is H or C 1-3 alkyl; and
RG is the maleimide reactive group.

4. A composition according to claim 2 , wherein said relative percentages are 70%-85% (i):5%-15% (ii):10%-15% (iii).

5. A composition according to claim 2 , wherein said relative percentages are about 75% (i), about 10% (ii), and about 15% (iii).

6. A composition according to claim 2 , wherein said sterol derivative has the following structure:

wherein:
one of R1 or R2 is —CH 2 —CH 2 —C(O)—RG and the other of R1 or R2 is H, or
R1 is —CH 2 —CH 2 —CH 2 —C(H)(CH 3 ) 2 and R2 is —C(O)—CH 2 —CH 2 —C(O)—RG,
wherein RG is the maleimide reactive group.

7. A composition according to claim 2 , wherein said liposomes are substantially between 50 and 500 nm in diameter.

8. A composition according to claim 7 , wherein said liposomes are substantially between 50 and 200 nm in diameter.

9. A composition according to claim 8 , wherein said liposomes are substantially between 50 and 150 nm in diameter.

10. A composition according to claim 2 , wherein said liposomes further comprise one or more components selected from the group consisting of monophosphoryl lipid A, resiquimod, flagellin, CpG, and α-galactosylceramide.

11. A composition according to claim 2 , wherein the lipid in (ii) is DMPG.

12. A composition according to claim 2 , wherein said sterol derivative(s) is/are derivative(s) of cholesterol, cholesteryl chloroformate, stigmasterol, sitosterol, ergosterol, lanosterol, desmosterol, or campesterol.

13. A composition according to claim 3 , wherein said relative percentages are 70%-85% (i):5%-15% (ii):10%-15% (iii).

14. A composition according to claim 3 , wherein said relative percentages are about 75% (i), about 10% (ii), and about 15% (iii).

15. A composition according to claim 3 , wherein said sterol derivative has the following structure:

wherein:
one of R1 or R2 is —CH 2 —CH 2 —C(O)—RG and the other of R1 or R2 is H, or
R1 is —CH 2 —CH 2 —CH 2 —C(H)(CH 3 ) 2 and R2 is —C(O)—CH 2 —CH 2 —C(O)—RG,
wherein RG is the maleimide reactive group.

16. A composition according to claim 3 , wherein said liposomes are substantially between 50 and 500 nm in diameter.

17. A composition according to claim 16 , wherein said liposomes are substantially between 50 and 200 nm in diameter.

18. A composition according to claim 17 , wherein said liposomes are substantially between 50 and 150 nm in diameter.

19. A composition according to claim 3 , wherein said liposomes further comprise one or more components selected from the group consisting of monophosphoryl lipid A, resiquimod, flagellin, CpG, and α-galactosylceramide.

20. A composition according to claim 3 , wherein the lipid in (ii) is DMPG.

21. A composition according to claim 3 , wherein said sterol derivative(s) is/are derivative(s) of cholesterol, cholesteryl chloroformate, stigmasterol, sitosterol, ergosterol, lanosterol, desmosterol, or campesterol.

Assignees

• Molecular Express Inc
• Univ California

Inventors

• Fujii Gary
• Szoka Jr Francis C
• Watson Douglas S

Classifications

• A61K2039/55555

  Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers · CPC title

• A61K2039/6018

  Lipids, e.g. in lipopeptides · CPC title

• C12N2760/16134

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• A61K39/145

  Orthomyxoviridae, e.g. influenza virus · CPC title

• A61K47/554

  the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid · CPC title