High purity lipopeptides

• US 8,129,342 B2
• Filed: 09/22/2010
• Issued: 03/06/2012
• Est. Priority Date: 01/20/2000
• Status: Expired due to Term

• Current Assignee: Cubist Pharmaceuticals, LLC (Merck & Co., Inc.)
• Original Assignee: Cubist Pharmaceuticals, Inc. (Merck & Co., Inc.)
• Inventors: Kelleher, Thomas J., Lai, Jan-Ji, DeCourcey, Joseph P., Lynch, Paul D., Zenoni, Maurizio, Tagliani, Auro R.
• Primary Examiner(s): Kam, Chih-Min
• Application Number:
  US12/888,233
• Publication Number:
  US 20110207658A1
• Time in Patent Office: 531 Days
• Field of Search: 514/9, 514/11, 514/2, 514/14, 530/317, 530/322, 530/344, 435/886
• US Class Current: 514/9
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 38/10 Peptides having 12 to 20 amino acids A61K38/043 - A61K38/046 take precedence
• CPC Class Codes: A61K 38/12 Cyclic peptides , e.g. bacitracins; Polymyxins; Gramicidins S, C; Tyrocidins A, B or C A61K38/043 - A61K38/046 take precedence
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: C07K 1/145 by extraction or solubilisation
• CPC Class Codes: C07K 5/0821 with the first amino acid being heterocyclic, e.g. His, Pro, Trp
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: Y10S 435/886 Streptomyces

Abstract

The invention discloses highly purified daptomycin and to pharmaceutical compositions comprising this compound. The invention discloses a method of purifying daptomycin comprising the sequential steps of anion exchange chromatography, hydrophobic interaction chromatography and anion exchange chromatography. The invention also discloses a method of purifying daptomycin by modified buffer enhanced anion exchange chromatography. The invention also discloses an improved method for producing daptomycin by fermentation of Streptomyces roseosporus. The invention also discloses high pressure liquid chromatography methods for analysis of daptomycin purity. The invention also discloses lipopeptide micelles and methods of making the micelles. The invention also discloses methods of using lipopeptide micelles for purifying lipopeptide antibiotics, such as daptomycin. The invention also discloses using lipopeptide micelles therapeutically.

Claims

1. A composition obtained by a process comprising the step of forming a daptomycin aggregate, the composition comprising daptomycin of greater than or about 93% purity relative to impurities 1-14 defined by peaks 1-14 shown in FIG. 12 and having less than 4% of anhydro-daptomycin and having less than 4% of β

-isomer of daptomycin.

2. The composition according to claim 1 that is free of each of impurities 1 to 14 defined by peaks 1-14 shown in FIG. 12.

3. The composition according to claim 1 that is essentially free of at least one of impurities 1 to 14 defined by peaks 1-14 shown in FIG. 12.

4. The composition of claim 1, wherein daptomycin purity is measured by HPLC.

5. The composition according to claim 1 wherein the process comprises the steps of:

i) providing a daptomycin solution under conditions in which the daptomycin is in a monomeric and nonmicellar state;

ii) filtering the daptomycin solution under conditions in which the daptomycin passes through the filter but pyrogens do not pass through the filter;

iii) subjecting the daptomycin solution to conditions forming a daptomycin aggregate;

iv) filtering the daptomycin aggregate under conditions in which the daptomycin aggregate is retained on the filter; and

v) collecting the daptomycin aggregate.

6. The composition according to claim 5, wherein the process further comprises the step of lyophilizing daptomycin.

7. The composition of claim 1 comprising daptomycin having less than 1% of β

-isomer of daptomycin.

8. The composition of claim 1, comprising daptomycin having greater than 93% purity.

9. The composition of claim 1, comprising daptomycin having less than 1% of the lactone hydrolysis product of daptomycin.

10. The composition of claim 1, comprising daptomycin that is substantially free of β

-isomer of daptomycin.

11. The composition of claim 10, comprising daptomycin of greater than 93% purity.

12. The composition of claim 1, comprising daptomycin of at least 95% purity.

13. The composition of claim 1, comprising daptomycin with a purity of about 94 to 96%.

14. The composition of claim 1, comprising daptomycin of at least 97% purity.

15. The composition of claim 1, comprising lyophilized daptomycin having greater than 93% purity relative to impurities 1-14 defined by peaks 1-14 shown in FIG. 12, and having less than 1% of the lactone hydrolysis product of daptomycin.

16. The composition of claim 15, wherein the daptomycin is substantially free of β

-isomer of daptomycin.

17. A pharmaceutical composition compatible with a pharmaceutically acceptable carrier for the treatment of an infection of the blood, skin or soft tissue, the composition comprising daptomycin obtained by a process comprising the step of forming a daptomycin aggregate, the composition having daptomycin with greater than or about 93% purity relative to impurities 1-14 defined by peaks 1-14 shown in FIG. 12.

18. The pharmaceutical composition of claim 17, wherein the daptomycin has greater than 93% purity and less than 4% anhydro daptomycin.

19. The pharmaceutical composition of claim 17, wherein the pharmaceutically acceptable carrier is selected from the group consisting of physiological saline and Ringer'"'"'s solution for reconstitution for administration as a single daily dose to the subject.

20. The pharmaceutical composition of claim 17, wherein the pharmaceutical composition is compatible with the pharmaceutically acceptable carrier for the treatment of an infection of the blood, skin or soft tissue by administration in a daily dose of 1 to 12 mg/kg of the daptomycin in a reconstituted solution of the composition in the pharmaceutically acceptable carrier.

21. The pharmaceutical composition of claim 20, wherein

a) the pharmaceutically acceptable carrier is selected from the group consisting of physiological saline and Ringer'"'"'s solution for intravenous administration as a single daily dose to the subject;

b) the daptomycin has greater than 93% purity, less than 4% anhydro daptomycin and less than 4% β

-isomer of daptomycin; and

c) the composition comprising daptomycin is obtained by a purification process comprising the steps of forming a daptomycin aggregate and obtaining the daptomycin from the daptomycin aggregate.

22. The pharmaceutical composition of claim 21, wherein the process for obtaining the daptomycin includes a purification process comprising the steps of

a) subjecting daptomycin to anion exchange chromatography to obtain an enriched daptomycin preparation;

b) forming the daptomycin aggregate comprising a daptomycin micelle in the enriched daptomycin preparation or a composition obtained from the enriched daptomycin preparation; and

c) obtaining the daptomycin from the daptomycin aggregate.

23. The pharmaceutical composition of claim 22, wherein the daptomycin is obtained from the daptomycin aggregate by a method comprising the steps of

a) filtering the daptomycin aggregate under conditions in which the daptomycin aggregate is retained on the filter;

and

b) collecting the daptomycin aggregate.

24. The pharmaceutical composition of claim 23, wherein the daptomycin is obtained from the daptomycin aggregate by a method further comprising the steps of

a) subjecting a composition comprising the daptomycin aggregate to hydrophobic interaction chromatography to obtain a semi-purified daptomycin preparation;

and

b) obtaining the daptomycin from the semi-purified daptomycin preparation.

25. The composition of claim 17, wherein the infection of the blood, skin or soft tissue comprises Staphylococcus aureus.

26. The composition of claim 17, wherein the infection is bacteremia.

27. The composition of claim 17, wherein the infection is endocarditis.

28. The composition of claim 17, wherein the infection is a skin or soft tissue infection.

29. The composition of claim 17, wherein the infection includes bacteria selected from the group consisting of Staphylococcus aureus, Streptococcus pyogenes, Streptococcus agalactiae, and Enterococcus faecalis.

30. A pharmaceutical composition for the treatment of an infection, the composition comprising daptomycin having greater than 93% purity relative to impurities 1-14 defined by peaks 1-14 shown in FIG. 12, the daptomycin purified by a process comprising the formation of micelles comprising daptomycin.

31. The pharmaceutical composition of claim 30, wherein the daptomycin is a lyophilized powder comprising daptomycin purified by process comprising the steps of forming a daptomycin micelle and obtaining the daptomycin from the micelles.

32. The pharmaceutical composition of claim 31, wherein the daptomycin has less than 4% anhydro daptomycin, less than 1% of the lactone hydrolysis product of daptomycin and is essentially free of the β

-isomer of daptomycin.

33. The pharmaceutical composition of claim 30, wherein the daptomycin has less than 4% anhydro daptomycin and less than 4% β

-isomer of daptomycin.

34. The pharmaceutical composition of claim 30, wherein the daptomycin has less than 1% of the lactone hydrolysis product of daptomycin.

35. The composition of claim 30, wherein the infection comprises Staphylococcus aureus.

36. The composition of claim 30, wherein the infection is bacteremia.

37. The composition of claim 30, wherein the infection is endocarditis.

38. The composition of claim 30, wherein the infection is a skin or soft tissue infection.

39. The composition of claim 30, wherein the infection includes bacteria selected from the group consisting of Staphylococcus aureus, Streptococcus pyogenes, Streptococcus agalactiae, and Enterococcus faecalis.

40. A pharmaceutical composition for the treatment of an infection of the blood, skin or soft tissue, the pharmaceutical composition comprising a solution of a pharmaceutically acceptable carrier for intravenous administration and daptomycin, the daptomycin having greater than 93% purity relative to impurities 1-14 defined by peaks 1-14 shown in FIG. 12, and the daptomycin obtained from a purification process comprising the formation of a daptomycin micelle.

41. The pharmaceutical composition of claim 40, wherein the daptomycin has less than 4% anhydro daptomycin and less than 4% β

-isomer of daptomycin.

42. The pharmaceutical composition of claim 41, wherein the daptomycin has less than 4% anhydro daptomycin, less than 1% of the lactone hydrolysis product of daptomycin and is essentially free of the β

-isomer of daptomycin.

43. The pharmaceutical composition of claim 40, wherein the daptomycin has less than 1% of the lactone hydrolysis product of daptomycin.

44. The composition of claim 40, wherein the infection comprises Staphylococcus aureus.

45. The composition of claim 40, wherein the infection is bacteremia.

46. The composition of claim 40, wherein the infection is endocarditis.

47. The composition of claim 40 wherein the infection is a skin or soft tissue infection.

48. The composition of claim 40, wherein the infection includes bacteria selected from the group consisting of Staphylococcus aureus, Streptococcus pyogenes, Streptococcus agalactiae, and Enterococcus faecalis.

49. The composition of claim 40, wherein the pharmaceutical composition includes daptomycin in a daily intravenous dose 1 to 12 mg/kg.

50. A vial containing a lyophilized powder pharmaceutical composition compatible with a pharmaceutically acceptable carrier for the treatment of an infection by a daily intravenous dose of the lyophilized powder reconstituted in the pharmaceutically acceptable carrier, the composition

a) having daptomycin with greater than or about 93% purity relative to impurities 1-14 defined by peaks 1-14 shown in FIG. 12;

and

b) the composition comprising daptomycin purified by a process including the steps of forming a daptomycin aggregate, converting the daptomycin aggregate to monomers and obtaining the daptomycin in the composition from the monomers by a process including one or more steps selected from the group consisting of anion exchange chromatography and hydrophobic interaction chromatography.

51. A composition obtained by a process comprising the steps of forming a daptomycin aggregate, converting the daptomycin aggregate to monomers and obtaining the daptomycin in the composition from the monomers by a process including one or more steps selected from the group consisting of anion exchange chromatography and hydrophobic interaction chromatography, the composition comprising daptomycin of greater than or about 93% purity relative to impurities 1-14 defined by peaks 1-14 shown in FIG. 12.

52. The composition of claim 51, comprising daptomycin of greater than 93% purity relative to impurities 1-14 defined by peaks 1-14 shown in FIG. 12.

53. The composition of claim 52, wherein

a) the composition is a lyophilized powder compatible with a pharmaceutically acceptable carrier for the treatment of an infection by a daily intravenous dose of 1 to 12 mg/kg of the daptomycin in a reconstituted solution of the lyophilized powder in the pharmaceutically acceptable carrier;

and

b) the daptomycin has a purity of about 94 to 96% relative to impurities 1-14 defined by peaks 1-14 shown in FIG. 12, the daptomycin having less than 1% of the lactone hydrolysis product of daptomycin, less than 4% anhydro daptomycin and less than 4% of the β

-isomer of daptomycin.

54. The composition of claim 51, wherein the composition is a lyophilized powder compatible with a pharmaceutically acceptable carrier for the treatment of one or more infections selected from the group consisting of infections of the blood, skin and soft tissue, by a daily intravenous dose of 1 to 12 mg/kg of the daptomycin in a reconstituted solution of the lyophilized powder in the pharmaceutically acceptable carrier.