Production of bupivacaine multivesicular liposomes

Abstract

Embodiments of this disclosure relate to a novel and improved large-scale commercial manufacturing process for producing bupivacaine multivesicular liposomes (MVLs). Batches of bupivacaine MVLs prepared by the novel process have high yield, improved stability, and a desired particle size distribution. Further embodiments of this disclosure relate to batches comprising a composition of bupivacaine-encapsulated multivesicular liposomes (MVLs) prepared by any one of the embodiments of the process described herein.

Claims

1. A batch comprising a composition of bupivacaine-encapsulated multivesicular liposomes (MVLs), wherein the composition comprises Bupivacaine present inside multiple internal aqueous chambers of an MVL separated by a lipid membrane, wherein the lipid membrane contains 1,2-dielycoilphosphatidylcholine (DEPC), 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG) or a salt thereof, cholesterol, and tricaprylin, and The aqueous medium in which the bupivacaine-encapsulated MVL is suspended, and also includes an aqueous medium containing unencapsulated bupivacaine. Includes, The total bupivacaine concentration in the composition is 12 mg / mL to 17 mg / mL. The batch has a volume of at least 100 liters, The batch, after storing two or more aliquots of the batch at 2°C to 8°C for approximately 12 months from the batch manufacturing date, is measured from the aliquots using an in vitro release assay assisted by a rotating apparatus for at least 48 hours, and has a cumulative release percentage of 46% to 71% bupivacaine at 24 hours. A batch in which the rate of change in the cumulative release percentage of bupivacaine in the batch at the 24-hour mark is at least 0.05% / month after the aliquots have been stored at 2°C to 8°C for approximately 12 months.

2. The batch according to claim 1, wherein the rate of change in the cumulative release percentage of bupivacaine at the 24-hour mark is 0.05% / month to 0.5% / month or 0.08% / month to 0.5% / month.

3. The batch according to claim 1 or 2, wherein the rate of change in the cumulative release percentage of bupivacaine at the 24-hour mark is 0.1% / month to 0.5% / month or 0.1% / month to 0.4% / month.

4. The batch according to any one of claims 1 to 3, wherein the batch has a cumulative release percentage of bupivacaine of 60% to 85% at 48 hours, and the rate of change in the cumulative release percentage of bupivacaine of the batch at 48 hours is not less than -0.3% per month after the aliquots are stored at 2°C to 8°C for about 12 months.

5. The batch according to claim 4, wherein the rate of change in the cumulative release percentage of bupivacaine at the 48-hour mark is -0.3% / month to 0.33% / month or -0.2% / month to 0.30% / month.

6. The batch according to claim 4 or 5, wherein the rate of change in the cumulative release percentage of bupivacaine at the 48-hour mark is -0.18% / month to 0.33% / month or -0.12% / month to 0.28% / month.

7. A batch comprising a composition of bupivacaine-encapsulated multivesicular liposomes (MVLs), wherein the composition comprises Bupivacaine present inside multiple internal aqueous chambers of an MVL separated by a lipid membrane, wherein the lipid membrane contains 1,2-dielycoilphosphatidylcholine (DEPC), 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG) or a salt thereof, cholesterol, and tricaprylin, and The aqueous medium in which the bupivacaine-encapsulated MVL is suspended, and also includes an aqueous medium containing unencapsulated bupivacaine. Includes, The total bupivacaine concentration in the composition is 12 mg / mL to 17 mg / mL. The batch has a volume of at least 100 liters, The batch, after storing two or more aliquots of the batch at 2°C to 8°C for approximately 12 months from the batch manufacturing date, is measured from the aliquots using an in vitro release assay assisted by a rotating apparatus for at least 48 hours, and has a cumulative release percentage of 60% to 85% of bupivacaine at 48 hours. A batch in which the rate of change in the cumulative release percentage of bupivacaine of the batch at the aforementioned 48-hour point is not less than -0.3% / month after the aliquots have been stored at 2°C to 8°C for approximately 12 months.

8. The batch according to claim 7, wherein the rate of change in the cumulative release percentage of bupivacaine at the 48-hour mark is -0.3% / month to 0.33% / month or -0.2% / month to 0.30% / month.

9. The batch according to claim 7 or 8, wherein the rate of change in the cumulative release percentage of bupivacaine at the 48-hour mark is -0.18% / month to 0.33% / month or -0.12% / month to 0.28% / month.

10. A batch comprising a composition of bupivacaine-encapsulated multivesicular liposomes (MVLs), wherein the composition comprises Bupivacaine present inside multiple internal aqueous chambers of an MVL separated by a lipid membrane, wherein the lipid membrane contains 1,2-dielycoilphosphatidylcholine (DEPC), 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG) or a salt thereof, cholesterol, and tricaprylin, and The aqueous medium in which the bupivacaine-encapsulated MVL is suspended, and also includes an aqueous medium containing unencapsulated bupivacaine. Includes, The total bupivacaine concentration in the composition is 12 mg / mL to 17 mg / mL. The batch is manufactured within a period of six months, and each of the batch has a volume of at least 100 liters. Each batch, after storing two or more aliquots from each batch at 2°C to 8°C for approximately 12 months from the batch manufacturing date, is measured for at least 48 hours using an in vitro release assay assisted by a rotating apparatus, and has a cumulative release percentage of 46% to 71% bupivacaine at 24 hours. A batch in which the average rate of change in the cumulative release percentage of bupivacaine in the batch at the 24-hour mark is at least 0.05% / month after storing the aliquots of each batch at 2°C to 8°C for approximately 12 months.

11. The batch according to claim 10, wherein the average rate of change in the cumulative release percentage of bupivacaine at the 24-hour mark is 0.05% / month to 0.5% / month or 0.08% / month to 0.5% / month.

12. The batch according to claim 10 or 11, wherein the average rate of change in the cumulative release percentage of bupivacaine at the 24-hour time point is 0.1% / month to 0.5% / month or 0.1% / month to 0.4% / month.

13. Each batch has a cumulative release percentage of bupivacaine of 60% to 85% at 48 hours, and the average rate of change in the cumulative release percentage of bupivacaine at 48 hours is not less than -0.3% / month, according to any one of claims 10 to 12.

14. The batch according to claim 13, wherein the average rate of change in the cumulative release percentage of bupivacaine at the 48-hour mark is -0.3% / month to 0.33% / month or -0.2% / month to 0.30% / month.

15. The batch according to claim 13 or 14, wherein the mean rate of change in the cumulative release percentage of bupivacaine at the 48-hour mark is -0.18% / month to 0.33% / month or -0.12% / month to 0.28% / month.

16. A batch comprising a composition of bupivacaine-encapsulated multivesicular liposomes (MVLs), wherein the composition comprises Bupivacaine present inside multiple internal aqueous chambers of an MVL separated by a lipid membrane, wherein the lipid membrane contains 1,2-dielycoilphosphatidylcholine (DEPC), 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG) or a salt thereof, cholesterol, and tricaprylin, and The aqueous medium in which the bupivacaine-encapsulated MVL is suspended, and also includes an aqueous medium containing unencapsulated bupivacaine. Includes, The total bupivacaine concentration in the composition is 12 mg / mL to 17 mg / mL. The batch is manufactured within a period of six months, and each of the batch has a volume of at least 100 liters. Each batch, after storing two or more aliquots of each batch at 2°C to 8°C for approximately 12 months from the batch manufacturing date, is measured from the aliquots of each batch using an in vitro release assay assisted by a rotating apparatus for at least 48 hours, and has a cumulative release percentage of 60% to 85% of bupivacaine at 48 hours. A batch in which the average rate of change in the cumulative release percentage of bupivacaine in the batch at the 48-hour mark is not less than -0.3% per month after storing the aliquots of each batch at 2°C to 8°C for approximately 12 months.

17. The batch according to claim 16, wherein the average rate of change in the cumulative release percentage of bupivacaine at the 48-hour mark is -0.3% / month to 0.33% / month or -0.2% / month to 0.30% / month.

18. The batch according to claim 16 or 17, wherein the mean rate of change in the cumulative release percentage of bupivacaine at the 48-hour mark is -0.18% / month to 0.33% / month or -0.12% / month to 0.28% / month.

19. The batch according to any one of claims 10 to 18, wherein the batch is manufactured within a period of three months.

20. The batch according to any one of claims 10 to 19, wherein the batch is manufactured within a period of 30 days.

21. The batch according to any one of claims 10 to 20, wherein the average rate of change in the cumulative release percentage of bupivacaine is such that, based on three batches, one batch is manufactured at least 10 days or longer apart from the other batches.

22. The batch according to any one of claims 10 to 20, wherein the average rate of change in the cumulative release percentage of bupivacaine is based on two batches, with one batch being manufactured 10 days or more apart from the other batch.

23. The batch or multiple batches according to any one of claims 1 to 22, wherein the cumulative release percentage of bupivacaine in each batch is measured as the average of six aliquots from each batch.

24. The batch or batch according to any one of claims 1 to 23, wherein each aliquot has a cumulative release percentage of bupivacaine of 36% to 81% at 24 hours and a cumulative release percentage of bupivacaine of 50% to 95% at 48 hours.

25. The batch or batch according to any one of claims 1 to 24, wherein the total bupivacaine concentration in the composition is about 13.3 mg / mL.

26. A batch or a number of batches according to any one of claims 1 to 25, wherein each batch has a volume of approximately 120 L to approximately 300 L, or approximately 150 L to approximately 280 L, or approximately 200 L to approximately 260 L.

27. below: (a) A step of mixing a first aqueous solution containing phosphoric acid with a volatile water-immiscible solvent solution to form a first water-in-oil emulsion, wherein the volatile water-immiscible solvent solution comprises 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG) or a salt thereof, 1,2-dielycoilphosphatidylcholine (DEPC), tricaprylin, and cholesterol, and either the first aqueous solution or the volatile water-immiscible solvent solution comprises bupivacaine. (b) A step of mixing the first water-in-oil emulsion with a second aqueous solution to form a second water-in-water emulsion, wherein the second aqueous solution contains lysine and at least one osmotic agent, (c) The step of substantially removing the volatile water-immiscible solvent from the second water-in-oil emulsion by sparging the second emulsion to form a first aqueous suspension of bupivacaine-encapsulated MVL having a first volume, (d) A step of reducing the first volume of the first aqueous suspension of bupivacaine-encapsulated polyvesicular liposomes by first microfiltration to provide a second aqueous suspension of bupivacaine-encapsulated MVL having a second volume, wherein the feed flow rate of the first microfiltration is about 200 L / min to about 400 L / min. (e) A step of replacing the medium of the second aqueous suspension with a saline solution by diafiltration to provide a third aqueous suspension of bupivacaine-filled MVL having a third volume, wherein the feed flow rate of the diafiltration is approximately 200 L / min to approximately 350 L / min. (f) The third volume of the third aqueous suspension is reduced by second microfiltration to provide a batch of aqueous suspensions of bupivacaine-encapsulated MVL having a target bupivacaine concentration of 12 mg / mL to 17 mg / mL. A batch according to any one of claims 1 to 9 and 23 to 26, prepared by a process including the following:

28. Each batch is as follows: (a) A step of mixing a first aqueous solution containing phosphoric acid with a volatile water-immiscible solvent solution to form a first water-in-oil emulsion, wherein the volatile water-immiscible solvent solution comprises 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG) or a salt thereof, 1,2-dielycoilphosphatidylcholine (DEPC), tricaprylin, and cholesterol, and either the first aqueous solution or the volatile water-immiscible solvent solution comprises bupivacaine. (b) A step of mixing the first water-in-oil emulsion with a second aqueous solution to form a second water-in-water emulsion, wherein the second aqueous solution contains lysine and at least one osmotic agent, (c) The step of substantially removing the volatile water-immiscible solvent from the second water-in-oil emulsion by sparging the second emulsion to form a first aqueous suspension of bupivacaine-encapsulated MVL having a first volume, (d) A step of reducing the first volume of the first aqueous suspension of bupivacaine-encapsulated polyvesicular liposomes by first microfiltration to provide a second aqueous suspension of bupivacaine-encapsulated MVL having a second volume, wherein the feed flow rate of the first microfiltration is about 200 L / min to about 400 L / min. (e) A step of replacing the medium of the second aqueous suspension with a saline solution by diafiltration to provide a third aqueous suspension of bupivacaine-filled MVL having a third volume, wherein the feed flow rate of the diafiltration is approximately 200 L / min to approximately 350 L / min. (f) The third volume of the third aqueous suspension is reduced by second microfiltration to provide a batch of aqueous suspensions of bupivacaine-encapsulated MVL having a target bupivacaine concentration of 12 mg / mL to 17 mg / mL. A batch according to any one of claims 10 to 26, prepared by a process including the following:

29. A process for preparing bupivacaine-encapsulated multivesicular liposomes (MVLs), (a) A step of mixing a first aqueous solution containing phosphoric acid with a volatile water-immiscible solvent solution to form a first water-in-oil emulsion, wherein the volatile water-immiscible solvent solution contains at least one phosphatidylcholine, at least one phosphatidylglycerol, cholesterol, and at least one neutral lipid, and either the first aqueous solution or the solvent solution contains bupivacaine. (b) A step of mixing the first water-in-oil emulsion with a second aqueous solution to form a second water-in-water emulsion, wherein the second aqueous solution contains lysine and at least one osmotic agent, (c) The step of substantially removing the volatile water-immiscible solvent from the second water-in-oil emulsion by sparging the second emulsion at a temperature of about 18°C ​​to about 20°C to form a first aqueous suspension of bupivacaine-encapsulated MVL having a first volume, (d) A step of reducing the first volume of the first aqueous suspension of bupivacaine-encapsulated polyvesicular liposomes by first microfiltration to provide a second aqueous suspension of bupivacaine-encapsulated MVL having a second volume, wherein the feed flow rate of the first microfiltration is at least 190 L / min. (e) A step of replacing the medium of the second aqueous suspension with a saline solution by diafiltration to provide a third aqueous suspension of bupivacaine-encapsulated MVL having a third volume, wherein the feed flow rate of the diafiltration is at least 190 L / min. (f) The third volume of the third aqueous suspension is reduced by second microfiltration to provide a final aqueous suspension of bupivacaine-encapsulated MVL having a target bupivacaine concentration of about 12 mg / mL to about 17 mg / mL. Includes, The final aqueous suspension of the bupivacaine-encapsulated MVL has a volume of at least 100 L. The process has a bupivacaine MVL product yield of at least about 75%. process.

30. The process according to claim 29, wherein the volatile water-immiscible solvent solution comprises bupivacaine, 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG) or a salt thereof, 1,2-dielucycoilphosphatidylcholine (DEPC), tricaprylin, and cholesterol.

31. The process according to claim 29 or 30, wherein the second aqueous solution comprises lysine and dextrose.

32. The process according to any one of claims 29 to 31, wherein the mixing in step (a) is carried out at a high speed of about 1100 rpm to about 1300 rpm for about 65 minutes to about 75 minutes.

33. The process according to claim 32, wherein the mixing in step (a) is carried out at a high speed of about 1200 rpm to about 1250 rpm for about 70 minutes.

34. The process according to any one of claims 29 to 31, wherein the mixing in step (a) is carried out at a high speed of about 1300 rpm to about 1500 rpm for about 70 minutes to about 90 minutes.

35. The process according to claim 34, wherein the mixing in step (a) is carried out at a high speed of about 1330 rpm to about 1350 rpm for about 70 minutes.

36. The process according to any one of claims 29 to 35, wherein the first water-in-oil emulsion has a volume of about 200 L to about 260 L.

37. The process according to any one of claims 29 to 36, wherein the mixing in step (b) is carried out at a low speed of about 445 rpm to about 680 rpm for about 60 to 85 seconds.

38. The process according to claim 37, wherein the mixing in step (b) is carried out at a low speed of about 615 rpm to about 650 rpm for about 70 seconds.

39. The process according to any one of claims 29 to 38, wherein the mixing in step (b) is carried out at a low speed of about 680 rpm to about 800 rpm for about 65 to 85 seconds.

40. The process according to claim 39, wherein the mixing in step (b) is carried out at a low speed of about 720 rpm to about 740 rpm for about 70 seconds.

41. The process according to any one of claims 29 to 40, wherein step (b) is carried out at a temperature of about 18°C ​​to about 20°C.

42. The process according to any one of claims 29 to 41, wherein the volume ratio of the first emulsion to the second emulsion is about 1:1 to 1:

5.

43. The process according to any one of claims 29 to 42, wherein the sparging of the second water-in-oil emulsion is carried out at a temperature of about 19°C for about 19 to about 25 minutes.

44. The process according to claim 43, wherein the sparging of the second water-in-oil emulsion comprises sparging for about 22 minutes at a sparging gas flow rate of about 1874 SLPM to about 2500 SLPM or a sparging gas flow rate of about 2400 SLPM.

45. The process according to any one of claims 29 to 44, wherein the first microfiltration is performed at a first microfiltration start feed flow rate of about 220 L / min to about 350 L / min and a first microfiltration end feed flow rate of about 190 L / min to about 310 L / min.

46. The process according to claim 45, wherein the feed flow rate of the first microfiltration decreases approximately linearly with respect to the MVL concentration in the first aqueous suspension, the starting feed flow rate of the first microfiltration is approximately 340 L / min, and the ending feed flow rate of the first microfiltration is approximately 300 L / min.

47. The process according to any one of claims 29 to 46, wherein the feed flow rate of the diafiltration during the first stage of the diafiltration is about 190 L / min to about 310 L / min, and the feed flow rate of the diafiltration during the second stage of the diafiltration is about 190 L / min to about 265 L / min.

48. The process according to claim 47, wherein the feed flow rate of the diafiltration during the first stage of the diafiltration is approximately 300 L / min, and the feed flow rate of the diafiltration during the second stage of the diafiltration is approximately 255 L / min.

49. The process according to any one of claims 29 to 48, wherein the second microfiltration is performed at a starting feed flow rate of approximately 190 L / min to approximately 265 L / min and a ending feed flow rate of approximately 120 L / min to approximately 190 L / min.

50. The process according to claim 49, wherein the feed flow rate of the second microfiltration decreases approximately linearly with respect to the MVL concentration in the third aqueous suspension, the starting feed flow rate of the second microfiltration is approximately 255 L / min, and the ending feed flow rate of the second microfiltration is approximately 180 L / min.

51. The process according to any one of claims 29 to 50, wherein steps (d), (e), and (f) are performed using a cross-flow filtration system configured to switch between modes between microfiltration and diafiltration, the cross-flow filtration system comprising a plurality of independently operating cross-flow modules, each cross-flow module comprising at least one filter array, and each filter array comprising a plurality of hollow fiber filters.

52. The process according to claim 51, wherein the cross-flow filtration system comprises two filter arrays, each filter array comprising six hollow fiber filters.

53. The process according to claim 51 or 52, wherein the cross-flow filtration system further includes at least one turbidity sensor downstream of the at least one filter array for detecting a loss of filter integrity.

54. A step of detecting an abnormal turbidity value using at least one turbidity sensor, wherein the abnormal turbidity value is a value exceeding a threshold value. A step of testing for possible filter integrity defects by sequentially deactivating one or more filters of the at least one filter array, and measuring the turbidity of each individual filter while it is deactivated using the at least one turbidity sensor until a normal turbidity value is detected, wherein the normal turbidity value is a value below the threshold, and The process according to claim 53, further comprising:

55. The process according to claim 54, further comprising the step of identifying the individual filters that were deactivated when the normal turbidity value was detected as potentially damaged filters, in response to the detection of a normal turbidity value.

56. The steps include: testing the potentially damaged filter by reactivating it and measuring the turbidity with at least one turbidity sensor while the potentially damaged filter is operating; In response to measuring an abnormal turbidity value when the potentially damaged filter is operating after restarting, the steps include determining that the potentially damaged filter is a damaged filter and stopping the operation of the damaged filter. The process according to claim 55, further comprising:

57. The process according to any one of claims 29 to 56, wherein the batch has a volume of approximately 120 L to approximately 300 L, approximately 150 L to approximately 280 L, approximately 200 L to approximately 260 L, approximately 320 L to approximately 500 L, or approximately 350 L to approximately 450 L.

58. The process according to any one of claims 29 to 57, having a bupivacaine MVL product yield of at least about 80%.

59. A composition of bupivacaine-encapsulated multivesicular liposomes (MVLs) prepared by the process described in any one of claims 29 to 57.

60. A batch comprising a composition of bupivacaine-encapsulated multivesicular liposomes (MVLs) prepared by the process described in any one of claims 29 to 57.

61. A method for treating or improving pain in a subject in need thereof, comprising the step of administering a composition according to any one of claims 1 to 28 and 59 to 60 to the subject.

62. The method according to claim 61, wherein the administration is by local infiltration into the surgical site to provide local analgesia.

63. The method according to claim 61, wherein the administration is by interscalene brachial plexus nerve block or femoral nerve block to provide regional analgesia.

64. The method according to claim 61, wherein the administration is by adductor canal block or sciatic nerve block in the popliteal fossa to provide regional analgesia.

65. The method according to any one of claims 61 to 64, wherein the composition has a volume of about 10 mL or 20 mL for a single dose.

Images