High and low load formulations of IGF-I in multivesicular liposomes

A process for producing multivesicular liposomes having multiple non-concentric chambers with membranes distributed as a continuous network throughout, and having insulin-like growth factor I (IGF-I) encapsulated therein, the process comprising the steps of:

• a. forming a first aqueous component comprising insulin-like growth factor I (IGF-I) in a concentration range from about 40 mg/mL to about 300 mg/mL, an osmotic excipient, and a pH adjusting agent, wherein the aqueous component is calibrated to have an osmolarity in the range from about 5 mOsm to about 150 mOsm and to have a pH in the range from about 2 to about 4.8;
• b. forming a lipid component comprising at least one organic solvent, at least one amphipathic lipid, and a neutral lipid lacking a hydrophilic headgroup;
• c. forming an emulsion from the first aqueous component and the lipid component;
• d. dispersing the emulsion into a second aqueous component to form solvent spherules;
• and
• e. removing the organic solvent from the solvent spherules to form multivesicular liposomes suspended in the second aqueous component.

A multivesicular liposome having multiple non-concentric chambers with membranes distributed as a continuous network throughout, made by a process comprising the steps of:

• a. forming a first aqueous component comprising insulin-like growth factor I (IGF-I) in a concentration range from about 1 mg/mL to about 33 mg/mL, an osmotic excipient and a pH adjusting agent, to have an osmolarity in the range from about 130 mOsm to about 320 mOsm and to have a pH in the range from about 1 to about 5;
• b. forming a lipid component comprising at least one organic solvent, at least one amphipathic lipid, and a neutral lipid lacking a hydrophilic headgroup;
• c. forming an emulsion from the first aqueous component and the lipid component;
• d. dispersing the emulsion into a second aqueous component to form solvent spherules;
• and
• e. removing the organic solvent from the solvent spherules to form multivesicular liposomes suspended in the second aqueous component.

A process for producing multivesicular liposome having multiple non-concentric chambers with membranes distributed as a continuous network throughout, and having insulin- like growth factor I (IGF-I) encapsulated therein, the process comprising the steps of:

• a. forming a first aqueous component comprising insulin-like growth factor I (IGF-I) in a concentration range from about 1 mg/mL to about 33 mg/mL, an osmotic excipient, and a pH adjusting agent, where the aqueous component is calibrated to have an osmolarity in the range from about 130 mOsm to about 320 mOsm and to have a pH in the range from about 1 to about 5, so as to determine resulting IGF-I load into the liposome;
• b. forming a lipid component comprising at least one organic solvent, at least one amphipathic lipid, and a neutral lipid lacking a hydrophilic headgroup;
• c. forming an emulsion from the first aqueous component and the lipid component;
• d. dispersing the emulsion into a second aqueous component to form solvent spherules;
• and
• e. removing the organic solvent from the solvent spherules to form multivesicular liposomes suspended in the second aqueous component.

A process for producing multivesicular liposome having multiple non-concentric chambers with membranes distributed as a continuous network throughout, and having insulin- like growth factor I (IGF-I) encapsulated therein, the process comprising the steps of:

• a. forming a first aqueous component comprising insulin-like growth factor I (IGF-I) in a concentration range from about 40 mg/mL to about 300 mg/mL, an osmotic excipient, and a pH adjusting agent, where the aqueous component is calibrated to have an osmolarity in the range from about 5 mOsm to about 150 mOsm and to have a pH in the range from about 2 to about 4.8, so as to determine resulting IGF-I load into the liposome;
• b. forming a lipid component comprising at least one organic solvent, at least one amphipathic lipid, and a neutral lipid lacking a hydrophilic headgroup;
• c. forming an emulsion from the first aqueous component and the lipid component;
• d. dispersing the emulsion into a second aqueous component to form solvent spherules;
• and
• e. removing the organic solvent from the solvent spherules to form multivesicular liposomes suspended in the second aqueous component;
• f. repeating steps a-e, except that once the desired IGF-I load is determined, exact amounts of IGF-I, osmotic excipient and pH and adjusting agent are used without further calibration.

A process for producing multivesicular liposome having multiple non-concentric chambers with membranes distributed as a continuous network throughout, and having insulin- like growth factor I (IGF-I) encapsulated therein, the process comprising the steps of:

• a. forming a first aqueous component comprising insulin-like growth factor I (IGF-I) in a concentration range from about 1 mg/mL to about 33 mg/mL, an osmotic excipient, and a pH adjusting agent, where the aqueous component is calibrated to have an osmolarity in the range from about 130 mOsm to about 320 mOsm and to have a pH in the range from about 1 to about 5;
• so as to determine resulting IGF-I load into the liposome;
• b. forming a lipid component comprising at least one organic solvent, at least one amphipathic lipid, and a neutral lipid lacking a hydrophilic headgroup;
• c. forming an emulsion from the first aqueous component and the lipid component;
• d. dispersing the emulsion into a second aqueous component to form solvent spherules;
• and
• e. removing the organic solvent from the solvent spherules to form multivesicular liposomes suspended in the second aqueous component;
• f. repeating steps a-e, except that once the desired IGF-I load is determined, exact amounts of IGF-I, osmotic excipient and pH and adjusting agent are used without further calibration.