Method for freeze-drying lipid nanoparticles
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- ARCTURUS THERAPEUTICS INC
- Filing Date
- 2026-03-19
- Publication Date
- 2026-06-23
Smart Images
Figure 2026102830000001_ABST
Abstract
Claims
1. A method for freeze-drying a composition containing lipid nanoparticles for encapsulating RNA, wherein the method is: a. A step of providing a suspension of lipid nanoparticles in a liquid medium, wherein the liquid medium contains approximately 4% w / v to approximately 22% w / v of saccharides; and b. The step of preparing the liquid medium to form a pre-treated suspension containing at least one excipient selected from potassium sorbate, thiosulfate, sodium benzoate, and iodixanol. The method, including the method described above.
2. Step (c): c. The pre-treated suspension i. An initial freezing process carried out at a temperature of -48 ± 8°C and atmospheric pressure; ii. A primary drying process carried out at a temperature in the range of -20±2°C to -48±2°C and a pressure in the range of approximately 25 mTorr to approximately 100 mTorr; and iii. A secondary drying process carried out at a temperature in the range of 5±2℃ to 30±2℃ and a pressure in the range of approximately 30 mTorr to approximately 300 mTorr. Subject to a freeze-thaw process that includes The method according to claim 1, further comprising:
3. Step (c): c. The pre-treated suspension i. An initial freezing process carried out at a temperature of -48 ± 8°C and atmospheric pressure; ii. A primary drying process carried out at a pressure of approximately 0.03 to approximately 0.08 mbar, starting at a temperature of -48 ± 8°C and gradually decreasing to a temperature of 0 ± 2°C over a period of approximately 40 to approximately 75 hours; and iii. A secondary drying process performed at a pressure of approximately 0.03 to 0.08 mbar, starting at a temperature of 0 ± 2°C and gradually increasing to a temperature of approximately 25 ± 3°C over a period of approximately 30 to 50 hours. Subject to a freeze-thaw process that includes The method according to claim 1, further comprising:
4. The method according to any one of claims 1 to 3, wherein the liquid medium is an aqueous medium.
5. The method according to any one of the prior claims, wherein the RNA in the suspension has a concentration in the range of about 0.05 mg / mL to about 2.0 mg / mL.
6. The method according to claim 5, wherein the RNA in the suspension has a concentration in the range of about 0.075 mg / mL to about 0.3 mg / mL.
7. The method according to claim 5, wherein the RNA in the suspension has a concentration in the range of about 0.1 mg / mL to about 1.5 mg / mL.
8. The method according to claim 5, wherein the RNA in the suspension has a concentration in the range of about 0.1 mg / mL to about 1.0 mg / mL.
9. The method according to claim 5, wherein the RNA in the suspension has a concentration in the range of about 0.1 mg / mL to about 0.5 mg / mL.
10. The method according to any one of the prior claims, wherein the weight ratio of total lipids to RNA in the suspension is about 50:1 to about 10:
1.
11. The method according to claim 10, wherein the weight ratio of total lipids to RNA in the suspension is about 40:1 to about 20:
1.
12. The method according to claim 10, wherein the weight ratio of total lipids to RNA in the suspension is about 35:1 to about 25:
1.
13. The method according to any one of the prior claims, wherein the pre-treated suspension comprises thiosulfate.
14. The method according to claim 13, wherein the thiosulfate is sodium thiosulfate or potassium thiosulfate.
15. The method according to claim 13, wherein the thiosulfate has a concentration of about 0.025% w / v to about 1.0% w / v.
16. The method according to claim 13, wherein the thiosulfate has a concentration of about 0.025% w / v to about 0.75% w / v.
17. The method according to claim 13, wherein the thiosulfate has a concentration of about 0.025% w / v to about 0.5% w / v.
18. The method according to claim 13, wherein the thiosulfate has a concentration of about 0.05% w / v to about 0.3% w / v.
19. The method according to claim 13, wherein the thiosulfate has a concentration of about 0.05% w / v to about 0.25% w / v.
20. The method according to any one of the prior claims, wherein the pre-treated suspension comprises potassium sorbate.
21. The method according to claim 20, wherein the potassium sorbate has a concentration of about 0.01 M to about 0.5 M.
22. The method according to claim 20, wherein the potassium sorbate has a concentration of about 0.02 M to about 0.4 M.
23. The method according to claim 20, wherein the potassium sorbate has a concentration of about 0.025 M to about 0.3 M.
24. The method according to claim 20, wherein the potassium sorbate has a concentration of about 0.03 M to about 0.2 M.
25. The method according to claim 20, wherein the potassium sorbate has a concentration of about 0.035 M to about 0.1 M.
26. The method according to claim 20, wherein the potassium sorbate has a concentration of about 0.04 M to about 0.08 M.
27. The method according to claim 20, wherein the potassium sorbate has a concentration of about 0.01 M to about 0.05 M.
28. The method according to claim 20, wherein the potassium sorbate has a concentration of about 0.02 M to about 0.04 M.
29. The method according to any one of the prior claims, wherein the pre-treated suspension comprises iodixanol.
30. The method according to claim 29, wherein the iodixanol has a concentration of about 5% w / v to about 15% w / v.
31. The method according to claim 29, wherein the iodixanol has a concentration of about 6% w / v to about 13% w / v.
32. The method according to claim 29, wherein the iodixanol has a concentration of about 7% w / v to about 11% w / v.
33. The method according to claim 29, wherein the iodixanol has a concentration of about 8% w / v to about 10% w / v.
34. The method according to any one of the prior claims, wherein the pre-treated suspension comprises sodium benzoate.
35. The method according to claim 34, wherein the sodium benzoate has a concentration of about 0.01 M to about 0.6 M.
36. The method according to claim 34, wherein the sodium benzoate has a concentration of about 0.02 M to about 0.5 M.
37. The method according to claim 34, wherein the sodium benzoate has a concentration of about 0.03 M to about 0.4 M.
38. The method according to claim 34, wherein the sodium benzoate has a concentration of about 0.04 M to about 0.3 M.
39. The method according to claim 34, wherein the sodium benzoate has a concentration of about 0.05 M to about 0.2 M.
40. The method according to any one of the prior claims, wherein the pre-treated suspension further comprises polyvinyl alcohol (PVA).
41. The method according to claim 40, wherein the PVA has a concentration of about 0.01% w / v to about 0.75% w / v.
42. The method according to any one of the prior claims, wherein the pre-treated suspension further comprises NaCl.
43. The method according to claim 42, wherein the NaCl has a concentration of about 0.005 M to about 0.5 M.
44. The method according to claim 42, wherein the NaCl has a concentration of about 0.01 M to about 0.4 M.
45. The method according to claim 42, wherein the NaCl has a concentration of about 0.015 M to about 0.3 M.
46. The method according to claim 42, wherein the NaCl has a concentration of about 0.015 M to about 0.2 M.
47. The method according to claim 42, wherein the NaCl has a concentration of about 0.015 M to about 0.1 M.
48. The method according to claim 42, wherein the NaCl has a concentration of about 0.02 M to about 0.05 M.
49. The method according to claim 42, wherein the NaCl has a concentration of about 0.03 M to about 0.07 M.
50. The method according to any one of the prior claims, wherein the saccharide is sucrose.
51. The method according to claim 50, wherein the sucrose has a concentration of about 8% w / v to about 20% w / v.
52. The method according to claim 50, wherein the sucrose has a concentration of about 7% w / v to about 11% w / v.
53. The method according to claim 50, wherein the sucrose has a concentration of about 8% w / v to about 10% w / v.
54. The method according to claim 50, wherein the sucrose has a concentration of about 16% w / v to about 20% w / v.
55. The method according to any one of the prior claims, comprising the liquid medium or pre-treated suspension, and a buffer.
56. The method according to claim 55, wherein the buffer solution is selected from MOPS, HEPES, Tris, MES, citrate, and phosphate-buffered saline (PBS).
57. The method according to claim 56, wherein the buffer solution is Tris.
58. The method according to any one of claims 55 to 57, wherein the buffer solution has a concentration of about 10 mM to about 100 mM.
59. The method according to claim 58, wherein the buffer solution has a concentration of about 15 mM to about 75 mM.
60. The method according to claim 58, wherein the buffer solution has a concentration of about 10 mM to about 40 mM.
61. The method according to any one of the prior claims, wherein the liquid medium or the pre-treated suspension has a pH of about 7.0 to about 8.
5.
62. The method according to any one of the prior claims, further comprising, after step (b), dividing the pre-treated suspension into individual containers in predetermined freeze-dried volumes.
63. The method according to claim 62, wherein the predetermined freeze-dried volume is in the range of about 0.5 mL to about 10.0 mL.
64. The method according to claim 62, wherein the predetermined freeze-dried volume is in the range of about 1.0 mL to about 3.0 mL.
65. The method according to any one of the prior claims, wherein the pre-treated suspension further comprises poloxamer.
66. The method according to claim 65, wherein the poloxamer is poloxamer 188.
67. The method according to claim 65 or claim 66, wherein the poloxamer is present in a concentration of about 0.01% w / v to about 0.10% w / v.
68. The method according to claim 65 or claim 66, wherein the poloxamer is present at a concentration of about 0.02% w / v to about 0.8% w / v.
69. The method according to claim 65 or claim 66, wherein the poloxamer is present at a concentration of about 0.03% w / v to about 0.7% w / v.
70. The method according to claim 65 or claim 66, wherein the poloxamer is present at a concentration of about 0.04% w / v to about 0.06% w / v.
71. A product prepared by the process described in any one of claims 1 to 70.
72. A lyophilized composition comprising lipid nanoparticles for encapsulating nucleic acids, monosaccharides, and one or more excipients selected from potassium sorbate, thiosulfate, sodium benzoate, and iodixanol.
73. The freeze-dried composition according to claim 72, wherein the nucleic acid is RNA.
74. The freeze-dried composition according to claim 73, wherein the RNA is self-replicating RNA.
75. The freeze-dried composition according to claim 73, wherein the RNA is mRNA.
76. The lyophilized composition according to any one of claims 73 to 75, wherein the nucleic acid has a length of about 20 nucleotides to about 13,000 nucleotides.
77. The freeze-dried composition according to any one of claims 72 to 76, wherein the weight ratio of total lipids to nucleic acids in the freeze-dried composition is about 50:1 to about 10:
1.
78. The freeze-dried composition according to claim 77, wherein the weight ratio of total lipids to RNA in the freeze-dried composition is about 40:1 to about 20:
1.
79. The freeze-dried composition according to claim 77, wherein the weight ratio of total lipids to RNA in the freeze-dried composition is about 35:1 to about 25:
1.
80. The freeze-dried composition according to any one of claims 72 to 79, wherein the freeze-dried composition contains potassium sorbate in a weight ratio of about 30:1 to about 250:1 of potassium sorbate to RNA.
81. The freeze-dried composition according to any one of claims 72 to 79, wherein the freeze-dried composition contains potassium sorbate in a weight ratio of about 40:1 to about 200:1 of potassium sorbate to RNA.
82. The freeze-dried composition according to any one of claims 72 to 79, wherein the freeze-dried composition contains potassium sorbate in a weight ratio of about 50:1 to about 175:1 of potassium sorbate to RNA.
83. The freeze-dried composition according to any one of claims 72 to 82, wherein the freeze-dried composition contains sodium thiosulfate in a weight ratio of sodium thiosulfate to RNA of about 0.25:1 to about 40:
1.
84. The freeze-dried composition according to any one of claims 72 to 82, wherein the freeze-dried composition contains sodium thiosulfate in a weight ratio of sodium thiosulfate to RNA of about 2:1 to about 10:
1.
85. The freeze-dried composition according to any one of claims 72 to 82, wherein the freeze-dried composition contains sodium thiosulfate in a weight ratio of sodium thiosulfate to RNA of about 3:1 to about 8:
1.
86. The freeze-dried composition according to any one of claims 72 to 85, wherein the freeze-dried composition contains sodium benzoate in a weight ratio of about 1:1 to about 12:1 of sodium benzoate to RNA.
87. The freeze-dried composition according to any one of claims 72 to 85, wherein the freeze-dried composition contains sodium benzoate in a weight ratio of about 2:1 to about 10:1 of sodium benzoate to RNA.
88. The freeze-dried composition according to any one of claims 72 to 85, wherein the freeze-dried composition contains sodium benzoate in a weight ratio of about 3:1 to about 9:1 of sodium benzoate to RNA.
89. The freeze-dried composition according to any one of claims 72 to 88, wherein the freeze-dried composition contains iodixanol in a weight ratio of iodixanol to RNA of about 100:1 to about 800:
1.
90. The freeze-dried composition according to any one of claims 72 to 88, wherein the freeze-dried composition contains iodixanol in a weight ratio of iodixanol to RNA of about 150:1 to about 750:
1.
91. The freeze-dried composition according to any one of claims 72 to 88, wherein the freeze-dried composition contains iodixanol in a weight ratio of iodixanol to RNA of about 200:1 to about 700:
1.
92. The freeze-dried composition according to any one of claims 72 to 88, wherein the freeze-dried composition contains iodixanol in a weight ratio of iodixanol to RNA of about 250:1 to about 650:
1.
93. The freeze-dried composition according to any one of claims 72 to 92, further comprising polyvinyl alcohol (PVA) in a weight ratio of PVA to RNA of about 1:1 to about 12:
1.
94. The freeze-dried composition according to any one of claims 72 to 93, wherein the saccharide is sucrose in a weight ratio of sucrose to RNA of about 100:1 to about 800:
1.
95. The lyophilized composition according to any one of claims 72 to 94, further comprising a buffer selected from HEPES, MOPS, Tris, MERS, citrate, and phosphate in a weight ratio of buffer to RNA of about 3:1 to about 150:
1.
96. A freeze-dried composition comprising lipid nanoparticles for encapsulating RNA, poloxamer, potassium sorbate, and sugar.
97. The composition according to claim 96, wherein the poloxamer is poloxamer 188.
98. The freeze-dried composition is the composition according to claim 96 or claim 97, comprising about 0.001 to about 1.0% w / w of the RNA.
99. The composition according to claim 98, wherein the freeze-dried composition contains about 0.005 to about 0.8% w / w of the RNA.
100. The composition according to claim 98, wherein the freeze-dried composition contains about 0.01 to about 0.5% w / w of the RNA.
101. The composition according to claim 98, wherein the freeze-dried composition contains about 0.02 to about 0.4% w / w of the RNA.
102. The composition according to claim 98, wherein the freeze-dried composition contains about 0.03 to about 0.3% w / w of the RNA.
103. The composition according to claim 98, wherein the freeze-dried composition contains about 0.04 to about 0.2% w / w of the RNA.
104. The freeze-dried composition is the composition according to any one of claims 96 to 103, comprising about 0.5 to about 5.0% w / w of lipids.
105. The freeze-dried composition according to claim 104, wherein the freeze-dried composition contains about 1.0 to about 4.0% w / w of lipids.
106. The composition according to claim 104, wherein the freeze-dried composition contains about 1.25 to about 3.0% w / w of lipids.
107. The lyophilized composition comprises about 0.5 to about 2.5% w / w of Tris buffer, according to any one of claims 96 to 106.
108. The composition according to claim 107, wherein the freeze-dried composition contains about 0.75 to about 2.25% w / w of Tris buffer.
109. The composition according to claim 107, wherein the freeze-dried composition contains about 1.0 to about 2.0% w / w of Tris buffer solution.
110. The freeze-dried composition is the composition according to any one of claims 96 to 109, comprising about 0.75 to about 2.75% w / w of NaCl.
111. The composition according to claim 110, wherein the freeze-dried composition contains about 1.0 to about 2.5% w / w of NaCl.
112. The freeze-dried composition according to claim 110, wherein the freeze-dried composition contains about 1.25 to about 1.80% w / w of NaCl.
113. The freeze-dried composition is the composition according to any one of claims 96 to 112, comprising about 85 to about 96% w / w of the sugar.
114. The composition according to claim 113, wherein the freeze-dried composition contains about 88 to about 95% w / w of the sugar.
115. The composition according to claim 113, wherein the freeze-dried composition contains about 90 to about 95% w / w of the sugar.
116. The composition according to any one of claims 96 to 115, wherein the sugar is sucrose.
117. The freeze-dried composition comprises about 0.01 to about 1.0% w / w of the poloxamer, according to any one of claims 96 to 116.
118. The composition according to claim 117, wherein the freeze-dried composition contains about 0.02 to about 0.8% w / w of the poloxamer.
119. The composition according to claim 117, wherein the freeze-dried composition contains about 0.03 to about 0.7% w / w of the poloxamer.
120. The composition according to claim 117, wherein the freeze-dried composition contains about 0.04 to about 0.6% w / w of the poloxamer.
121. The composition according to claim 117, wherein the freeze-dried composition contains about 0.05 to about 0.5% w / w of the poloxamer.
122. The composition according to claim 117, wherein the freeze-dried composition contains about 0.06 to about 0.4% w / w of the poloxamer.
123. The composition according to claim 117, wherein the freeze-dried composition contains about 0.07 to about 0.3% w / w of the poloxamer.
124. The composition according to claim 117, wherein the freeze-dried composition contains about 0.09 to about 0.2% w / w of the poloxamer.
125. The composition according to any one of claims 96 to 124, wherein the poloxamer is poloxamer 188.
126. The freeze-dried composition is the composition according to any one of claims 96 to 125, comprising about 0.5 to about 5.0% w / w of potassium sorbate.
127. The composition according to claim 126, wherein the freeze-dried composition contains about 0.75 to about 4.0% w / w of potassium sorbate.
128. The composition according to claim 126, wherein the freeze-dried composition contains about 1.0 to about 3.0% w / w of potassium sorbate.
129. The composition according to claim 126, wherein the freeze-dried composition contains about 1.25 to about 2.75% w / w of potassium sorbate.
130. A method for storing a freeze-dried composition according to any one of claims 72 to 129, comprising storing the freeze-dried product at a temperature of about 2°C to about 8°C.
131. A method for storing a freeze-dried composition according to any one of claims 72 to 129, comprising storing the freeze-dried product at a temperature of about -20°C.
132. A method for reconstituting a freeze-dried composition according to any one of claims 72 to 129, comprising adding a liquid medium to the freeze-dried composition.
133. The method according to claim 132, wherein the liquid medium is an aqueous medium.
134. The method according to claim 132 or claim 133, wherein the liquid medium comprises poloxamer.
135. The method according to claim 134, wherein the poloxamer is P-188.
136. The method according to any one of claims 132 to 135, wherein the liquid medium further comprises a buffer having a pH of about 7.0 to about 8.
5.
137. A method for treating a disease or disorder in a subject, comprising administering to the subject a lyophilized composition according to any one of claims 72 to 129, which has been reconstituted in a liquid medium.
138. The method according to claim 137, wherein the reconstituted lyophilized composition is administered intravenously.
139. The method according to claim 137, wherein the reconstituted lyophilized composition is administered intramuscularly.
140. The method according to claim 137, wherein the reconstituted lyophilized composition is administered by inhalation.
141. The method according to claim 137, wherein the reconstituted freeze-dried composition is administered to a mucous membrane.
142. The method according to claim 137, wherein the reconstituted lyophilized composition is administered subcutaneously.