Methods of evaluating protein formulation stability and surfactant-stabilized insulin formulations derived therefrom

• US 20030054979A1
• Filed: 10/18/2001
• Published: 03/20/2003
• Est. Priority Date: 06/28/2001
• Status: Active Grant

• Current Assignee: Medtronic Minimed Incorporated (Medtronic Plc)
• Original Assignee: Medtronic Minimed Incorporated (Medtronic Plc)
• Inventors: Gulati, Poonam S., Van Antwerp, William P., Gross, Todd M., Kim, Seonyoung
• Granted Patent: US 6,737,401 B2
• Time in Patent Office: Days
• US Class Current: 514/4
• CPC Class Codes: A61K 47/02 Inorganic compounds
• CPC Class Codes: A61K 47/10 Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
• CPC Class Codes: A61K 47/18 Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
• CPC Class Codes: A61K 47/26 Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61M 5/14 Infusion devices, e.g. infusing by gravity; Blood infusion; Accessories therefor
• CPC Class Codes: G01N 33/6803 General methods of protein analysis not limited to specific proteins or families of proteins
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors

Abstract

Embodiments of the invention are directed to a method of estimating the physical stability of a protein formulation. A particular embodiment of the invention places the protein formulation under an agitational stress that causes the protein to aggregate at an accelerated rate. In one embodiment, the change in protein aggregation is monitored spectroscopically using Thioflavin-T. Embodiments of the invention then utilize a survival curve analysis to ascertain the relative physical stability of the different protein formulations under study. This method was used to develop novel surfactant-stabilized insulin formulations in a rapid, cost efficient manner, thus illustrating the utility of the inventive method to the discovery and development of pharmaceutical protein formulations.

Claims

1. An aqueous insulin formulation comprising insulin, a buffer system, an isotonicity agent, a preservative, metal ions, and a non-ionic surfactant selected from at least a polysorbate, a poloxyethylene ether, a polyethylene glycol ether, and mixtures of these surfactants.

2. The aqueous insulin formulation of claim 1, wherein the insulin is human insulin.

3. The aqueous insulin formulation of claim 2, wherein the human insulin is a recombinant insulin.

6. The aqueous insulin formulation of claim 1, wherein the insulin concentration is about 2 U/ml to about 1000 U/ml.

7. The aqueous insulin formulation of claim 6, wherein the insulin concentration is about 400 U/ml.

4. The aqueous insulin formulation, wherein the preservative is selected from at least phenol, m-cresol and mixture of these preservatives.

5. An aqueous insulin formulation comprising insulin, a buffer system, an isotonicity agent, a preservative, metal ions and a non-ionic surfactant selected from at least a polysorbate, a poloxyethylene ether, a polyethylene glycol, and mixtures of these surfactants, wherein the insulin formulation exhibits a physical stability that is greater than or equal to the same insulin formulation containing the non-ionic surfactant Genapol PF-10.

8. An insulin infusion device comprising:

an insulin pump system; and

a surfactant-stabilized insulin formulation including insulin and a non-ionic surfactant selected from at least a polysorbate, a poloxyethylene ether, a polyethylene glycol ether, and mixtures of these surfactants.

9. A method of evaluating the physical stability of a protein formulation comprising:

preparing a statistically relevant number of identical samples of a protein formulation to yield a first sample type, wherein the protein is susceptible to changes in its native conformation yielding non-native conformers of the protein;

preparing a statistically relevant number of identical samples of at least one other protein formulation that differs from the first sample type to yield a second, or more, sample types, wherein the protein is susceptible to a changes in its native conformation yielding non-native conformers of the protein;

adding an agent that yields a change upon binding to a non-native conformer of the protein;

applying a controlled stress on all sample types, wherein the controlled stress applied causes the protein to exhibit a change in its native conformation;

monitoring the samples types to yield time-dependent data that are related to a degree of protein conformational change for each sample type;

applying a survival analysis to the data obtained for each sample type; and

comparing the survival analysis for each sample type to determine the relative physical stability of the protein formulations under evaluation.

10. The method of claim 9, wherein the controlled stress is agitation.

11. The method of claim 9, wherein the spectroscopic change is a spectroscopic change in fluorescence.

12. The method of claim of claim 9, wherein the protein is insulin and the non-native conformer of the protein is a fibril form of insulin.