A single-chain insulin analogues may comprise an insulin B-chain polypeptide sequence connected by a connecting polypeptide (or C-domain) sequence to an insulin A-chain polypeptide sequence. The connecting polypeptide sequence may be Glu-Xaa-Gly-Pro-Arg-Arg where Xaa is Glu or Ala. The insulin analogues may additionally comprise Glu or His substitutions at the position corresponding to A8 of human insulin and/or a Glu substitution at the position corresponding to A14 of human insulin. In some embodiments, the insulin analogues may additionally comprise either a Pro or Glu at the positions corresponding to B28 and B29 of wild-type insulin. Additional substitutions may comprise Phe or Trp at the position corresponding to A13 of wild type insulin and/or Gln, Arg, Phe, or Glu at the position corresponding to A17 of wild type insulin. In some embodiments, a Glu substitution at the position corresponding to B16 of wild type insulin may be present. In other embodiments, a Cys substitution may be present at the positions corresponding to A10 and/or B4 of wild-type insulin. In addition or in the alternative, the analogue may comprise a His or Ala substitution at the position corresponding to B22 of wild-type insulin and/or the connecting polypeptide sequence may be Glu-Glu-Gly-Pro-Ala-His. A method of treating a patient with diabetes mellitus comprises administering a physiologically effective amount of the insulin analogue or a physiologically acceptable salt thereof to a patient by means of intravenous, intraperitoneal, or subcutaneous injection.La présente invention concerne un analogue d'insuline à chaîne unique qui peut comprendre une séquence polypeptidique à chaîne B d'insuline reliée par une séquence polypeptidique de liaison (ou de domaine C) à une séquence polypeptidique à chaîne A d'insuline. La séquence polypeptidique de liaison peut être Glu-Xaa-Gly-Pro-Arg-Arg dans laquelle Xaa est Glu ou Ala. Les analogues d'insuline peuvent en outre comprendre des substitutio
