Biphasic single-chain insulin analogues

A pharmaceutical composition comprising a single-chain insulin formulated at a pH within the range 7.0 to 8.0, wherein the single chain insulin comprises:

• a C-domain of from 6 to 11 amino acid residues comprising at least two acidic residues at the N-terminal side of the C-domain and at least two basic residues at the C-terminal side of the C-domain peptide;
• an amino acid substitution at the position corresponding to A8 of human insulin selected from the group consisting of Ala, Gln, and Glu, and
• an amino acid substitution at the position corresponding to A14 of human insulin selected from the group consisting of Gln, Phe and Trp.

A method for lowering the blood sugar level of a patient in need thereof, the method comprising, subcutaneously administering a pharmaceutical composition to the patient, wherein the pharmaceutical composition comprises a single-chain insulin formulated at a pH within the range 7.0 to 8.0, and wherein the single chain insulin comprises:

• a C-domain of from 6 to 11 amino acid residues comprising at least two acidic residues at the N-terminal side of the C-domain and at least two basic residues at the C-terminal side of the C-domain peptide;
• an amino acid substitution at the position corresponding to A8 of human insulin selected from the group consisting of Ala, Gln, and Glu, and
• an amino acid substitution at the position corresponding to A14 of human insulin selected from the group consisting of Gln, Phe and Trp.