Discloses insulin analogues with the general structure as shown in Formula 1 with amino acid substitutions in the A-chain and B-chain as disclosed. Further discloses a method for preparing the insulin analogues. Also discloses insulin analogues which further comprise an addition of a glucagon-like peptide-1 or extendin-3 or -4 molecule. The therapeutic application is in the treatment of diabetes by providing insulin analogues with long-lasting activity in vivo.