Compounds for protein stabilization and methods for their use

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.

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.

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.

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.

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.

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.

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.