Compounds for protein stabilization and methods for their use

• US 20040171518A1
• Filed: 02/27/2003
• Published: 09/02/2004
• Est. Priority Date: 02/27/2003
• Status: Abandoned Application

• Current Assignee: Medtronic Minimed Incorporated (Medtronic Plc)
• Original Assignee: Medtronic Minimed Incorporated (Medtronic Plc)
• Inventors: Walsh, Joseph C., Van Antwerp, William Peter
• Application Number:
  US10/375,383
• Publication Number:
  US 20040171518A1
• Time in Patent Office: Days
• US Class Current: 514/3
• 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 31/495 having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazinesA61K31/48 takes precedence ; with three nitrogen atoms A61K31/53
• CPC Class Codes: A61K 38/28 Insulins
• CPC Class Codes: A61K 47/22 Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
• 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

Abstract

The present invention is directed to stabilized polypeptide compositions. Typical embodiments of the present invention provide improved methods and materials for maintaining the stability of insulin polypeptide formulations. In particular, the disclosure provided herein teaches that the aggregation of insulin polypeptides can be inhibited by combining them with a class of compounds having the general formula A¹—L¹—S—L²—A², wherein S comprises from one to seven consecutive atoms selected from the group consisting of nitrogen, carbon, and oxygen, wherein at least one of the atoms is a carbon atom; L¹ and L² are linking groups having from two to twelve atoms selected from the group consisting of nitrogen, carbon, oxygen, sulfur, and phosphorus; and A¹ and A² are carboxylic acid groups.

Claims

1. A method of inhibiting the aggregation of insulin polypeptides in a solution comprising combining the insulin polypeptides with a compound having the general formula:

2. The method of claim 1, wherein the compound comprises a compound having a general formula shown in FIG. 2.

3. The method of claim 2, wherein the compound has the general formula:

4. The method of claim 1, wherein S has the structure:

5. The method of claim 1, wherein L¹ and L² have the structure:

6. The method of claim 1, wherein A¹ and A² have the structure:

7. The composition of claim 1, wherein insulin polypeptide is LISPRO insulin.

8. The composition of claim 1, wherein the composition is a pharmaceutical composition.

9. The method of claim 1, wherein the solution comprises zinc.

10. The method of claim 1, wherein the compound further comprises a phenolic moiety.

11. The method of claim 1, wherein the inhibition of aggregation is observable in a spectrophotometric assay of turbidity or a Thioflavin-T assay.

12. A method of increasing the stability of insulin polypeptides in a solution comprising combining the insulin polypeptides with a compound having the general formula:

A¹—

L¹—

S—

L²—

A²

wherein S comprises from one to seven atoms selected from the group consisting of nitrogen, carbon, and oxygen, wherein at least one of the atoms is a carbon atom;

L¹ and L² are linking groups having from two to twelve consecutively linked atoms selected from the group consisting of nitrogen, carbon, oxygen, sulfur, and phosphorus; and

A¹ and A² ate carboxylic acid groups; and

wherein the amount of compound in the solution is sufficient to inhibit the aggregation of the insulin polypeptides in the solution.

13. The method of claim 12, wherein the compound has the general formula:

14. A method of producing a stabilized insulin solution comprising combining insulin with a compound having the general formula:

A¹—

L¹—

S—

L²—

A²

wherein S comprises from one to seven atoms selected from the group consisting of nitrogen, carbon, and oxygen, wherein at least one of the atoms is a carbon atom;

L¹ and L² are linking groups having from two to twelve consecutively linked atoms selected from the group consisting of nitrogen, carbon, oxygen, sulfur, and phosphorus; and

A¹ and A² are carboxylic acid groups; and

wherein the compound inhibits the aggregation of the insulin in the solution.

15. The method of claim 14, wherein the compound has the general formula:

16. A method of making a compound for use in inhibiting the aggregation of insulin polypeptides in solution comprising:

(a) constructing a compound having the general formula;

A¹—

L¹—

S—

L²—

A²

wherein S comprises from one to seven atoms selected from the group consisting of nitrogen, carbon, and oxygen, wherein at least one of the atoms is a carbon atom;

L¹ and L² ate linking groups having from two to twelve consecutively linked atoms selected from the group consisting of nitrogen, carbon, oxygen, sulfur, and phosphorus; and

A¹ and A² are carboxylic acid groups; and

(b) testing the compound in an assay of insulin polypeptide aggregation, wherein the level of insulin polypeptide aggregation that is observed in a solution comprising insulin polypeptides and the compound is less than that observed in a control sample to which no compound has been added.

17. A method of identifying a compound for use in inhibiting the aggregation of insulin polypeptides in solution comprising combining insulin polypeptides with a compound having the general formula:

A¹—

L¹—

S—

L²—

A²

wherein S comprises from one to seven atoms selected from the group consisting of nitrogen, carbon, and oxygen, wherein at least one of the atoms is a carbon atom;

L¹ and L² are linking groups having from two to twelve consecutively linked atoms selected from the group consisting of nitrogen, carbon, oxygen, sulfur, and phosphorus; and

A¹ and A² are carboxylic acid groups; and

(b) characterizing the compound in an assay of insulin polypeptide aggregation, wherein the level of insulin polypeptide aggregation that is observed in the solution comprising the insulin polypeptides and the compound is less than that observed in a control sample to which no compound has been added.

18. A method of inhibiting the generation of an immune response to exogenous insulin comprising administering the exogenous insulin in a solution combination with a compound having the general formula:

A¹—

L¹—

S—

L²—

A²

wherein S comprises from one to seven atoms selected from the group consisting of nitrogen, carbon, and oxygen, wherein at least one of the atoms is a carbon atom;

L¹ and L² are linking groups having from two to twelve consecutively linked atoms selected from the group consisting of nitrogen, carbon, oxygen, sulfur, and phosphorus; and

A¹ and A² are carboxylic acid groups;

wherein the amount of compound combined with the insulin polypeptides is sufficient to inhibit the aggregation of the insulin polypeptides in the solution.

19. The method of claim 18, wherein the compound has the general formula:

20. A method of inhibiting the formation of immunoreactive insulin complexes in a solution comprising combining insulin in a solution a compound having the general formula:

A¹—

L¹—

S—

L²—

A²

wherein S comprises from one to seven atoms selected from the group consisting of nitrogen, carbon, and oxygen, wherein at least one of the atoms is a carbon atom;

L¹ and L² are linking groups having from two to twelve consecutively linked atoms selected from the group consisting of nitrogen, carbon, oxygen, sulfur, and phosphorus; and

A¹ and A² are carboxylic acid groups;

wherein the amount of compound combined with the insulin polypeptides is sufficient to inhibit the aggregation of the insulin polypeptides in the solution.

21. The method of claim 20, wherein the compound has the general formula:

22. A method of inhibiting the aggregation of polypeptides in a solution comprising combining the polypeptides with a compound having the general formula:

A¹—

L¹—

S—

L²—

A²

wherein S comprises from one to seven atoms selected from the group consisting of nitrogen, carbon, and oxygen, wherein at least one of the atoms is a carbon atom;

L¹ and L² are linking groups having from two to twelve consecutively linked atoms selected from the group consisting of nitrogen, carbon, oxygen, sulfur, and phosphorus; and

A¹ and A² are carboxylic acid groups;

wherein the amount of compound combined with the polypeptides is sufficient to inhibit the aggregation of the polypeptides in the solution.

23. The method of claim 22, wherein the polypeptide is LISPRO insulin.

24. The method of claim 22, wherein the compound has the general formula:

25. A compound having a general formula shown in FIG. 2.