Methods for preparing squalene

Patent metadata

Field	Value
Publication number	US-9545440-B2
Application number	US-201514921665-A
Country	US
Kind code	B2
Filing date	Oct 23, 2015
Priority date	May 12, 2010
Publication date	Jan 17, 2017
Grant date	Jan 17, 2017

Abstract

An improved method for preparing squalene from a squalene-containing composition, said method comprising the steps of (a) a purification distillation carried out at a temperature T 1 (b) a denaturing distillation carried out at a temperature T 2 ; wherein steps (a) and (b) may be performed in either order; T 1 and T 2 are sufficient to cause squalene to boil; T 2 >T 1 ; and T 2 >200° C.

First claim

The invention claimed is:

1. A method for the manufacture of an oil-in-water emulsion comprising:
i. obtaining squalene prepared from a fish oil source by a process comprising subjecting fish oil comprising squalene to: (a) a purification distillation carried out at near vacuum; and (b) a denaturing distillation carried out at a temperature greater than or equal to 210° C. at a pressure of 0.8 to 5.0 mmHg,
wherein the temperature in the denaturing distillation exceeds the temperature in the purification distillation; and
wherein the conditions for both the purification and the denaturing distillation are sufficient to cause squalene to boil; and
wherein the denaturing distillation denatures and/or removes potential contaminant proteins and viruses, thereby rendering the squalene safe for human use as compared with non-denatured squalene; and

ii. preparing an oil-in-water emulsion using the squalene from step (i).

2. The method of claim 1 , wherein the purification distillation occurs at a temperature of less than 140° C.

3. The method of claim 1 , wherein the squalene prepared in step (i) is kept sterile following distillation treatment and prior to the preparation of the oil-in-water emulsion.

4. The method of claim 1 , wherein the fish oil comprises one or more contaminant proteins.

5. The method of claim 4 , wherein the contaminant protein comprises parvalbumin.

6. The method of claim 1 , wherein the purification distillation is carried out at a temperature of from 70 to 100° C.

7. The method of claim 1 , wherein the purification distillation is carried out at a pressure of from 0.5 μm Hg to 5 μm Hg.

8. The method of claim 1 , wherein the purification distillation is carried out prior to the denaturing distillation.

9. The method of claim 1 , which further comprises a step of subjecting the fish oil to saponification prior to conducting the purification distillation and denaturing distillation steps.

10. The method of claim 1 , which further comprises a step of subjecting the fish oil to saponification before conducting the denaturing distillation step.

11. The method of claim 9 , wherein saponification comprises the addition of NaOH or KOH to the fish oil.

12. The method of claim 10 , wherein saponification comprises the addition of NaOH or KOH to the fish oil.

13. The method of claim 1 , wherein the purification distillation results in a composition comprising 99% squalene or more.

14. The method of claim 1 , wherein the fish oil is shark liver oil.

15. The method of claim 1 , further comprising the step of combining the oil-in-water emulsion with an antigen.

16. The method of claim 1 , further comprising the step of packaging the oil-in-water emulsion into a kit as a kit component together with an antigen component.

17. The method of claim 16 , wherein the kit components are in separate vials.

18. The method of claim 17 , wherein the vials are made from borosilicate glass.

19. The method of claim 15 , wherein the antigen is an influenza virus antigen.

20. The method of claim 19 , wherein the combination of the oil-in-water emulsion and the antigen forms a vaccine composition and wherein the vaccine composition includes about 15 μg; about 10 μg, about 7.5 μg, about 5 μg, about 3.8 μg, about 3.75 μg, about 1.9 μg, or about 1.5 μg of hemagglutinin per influenza virus strain.

21. The method of claim 19 , wherein the combination of the oil-in-water emulsion and the antigen forms a vaccine composition and wherein the vaccine composition includes a thiomersal or 2-phenoxyethanol preservative.

22. The method of claim 1 , wherein the oil-in-water emulsion comprises between 2-10% squalene.

23. The method of claim 1 , wherein the oil-in-water emulsion comprises squalene, polysorbate 80 and sorbitan trioleate.

24. A method for processing squalene from a fish source, said method comprising the steps of:
i providing a composition comprising distilled squalene which has been obtained by a process of distillation, and wherein the squalene is from a fish source; and
ii re-distilling the composition from (i) at a temperature sufficient to cause the distilled squalene to boil, so as to provide a composition comprising re-distilled squalene;
wherein the process of distillation is performed at a temperature less than 140° C. and at a near vacuum and the process of re-distillation is performed at a temperature greater than or equal to 210° C. and a pressure of 0.8 to 5.0 mmHg; and
wherein the conditions for both the distillation and the re-distillation are sufficient to cause the squalene to boil; and
wherein the denaturing distillation denatures and/or removes potential contaminant proteins and viruses, thereby rendering the squalene safer for human use as compared with non-denatured squalene.

25. A method for the manufacture of an oil-in-water emulsion comprising:
i. obtaining squalene prepared from a fish oil source by a process comprising subjecting fish oil comprising squalene to: (a) a purification distillation carried out at near vacuum at a temperature of less than 140° C.; and (b) a denaturing distillation carried out at a temperature greater than or equal to 210° C.,
wherein the conditions for both the purification and the denaturing distillation are sufficient to cause squalene to boil; and
wherein the denaturing distillation denatures and/or removes potential contaminant proteins and viruses, thereby rendering the squalene safe for human use as compared with non-denatured squalene; and

ii. preparing an oil-in-water emulsion using the squalene from step (i).

26. The method of claim 25 , wherein the squalene prepared in step (i) is kept sterile following distillation treatment and prior to the preparation of the oil-in-water emulsion.

27. The method of claim 25 , wherein the fish oil comprises one or more contaminant proteins.

28. The method of claim 27 , wherein the contaminant protein comprises parvalbumin.

29. The method of claim 25 , wherein the purification distillation is carried out at a temperature of from 70 to 100° C.

30. The method of claim 25 , wherein the purification distillation is carried out at a pressure of from 0.5 μm Hg to 5 μm Hg.

31. The method of claim 25 , wherein the purification distillation is carried out prior to the denaturing distillation.

32. The method of claim 25 , which further comprises a step of subjecting the fish oil to saponification prior to conducting the purification distillation and denaturing distillation steps.

33. The method of claim 25 , which further comprises a step of subjecting the fish oil to saponification before conducting the denaturing distillation step.

34. The method of claim 32 , wherein saponification comprises the addition of NaOH or KOH to the fish oil.

35. The method of claim 33 , wherein saponification comprises the addition of NaOH or KOH to the fish oil.

36. The method of claim 25 , wherein the purification distillation results in a composition comprising 99% squalene or more.

37. The method of claim 25 , wherein the fish oil is shark liver oil.

38. The method of claim 25 , further comprising the step of combining the oil-in-water emulsion with an antigen.

39. The

Assignees

• Novartis Ag

Inventors

• Hora Maninder

Classifications

• A61P31/00

  Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics · CPC title

• A61P31/16

  for influenza or rhinoviruses · CPC title

• B01L3/10

  Wash bottles · CPC title

• C12N2760/16034

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

• A61K9/107

  Emulsions {; Emulsion preconcentrates; Micelles (composition of emulsions A61K47/00)} · CPC title