The invention discloses a
solid-
phase synthesis insulin glargine preparation process, which belongs to the technical field of biological
pharmacy, and comprises the following steps: S1, carrying out
solid-
phase synthesis on an
insulin A chain through
Wang resin, removing a
protecting group, and then oxidizing by using NCS to form a first pair of disulfide bonds: sequentially inoculating amino acids according to an Fmoc
solid-
phase synthesis method by using NCS as an
oxidizing agent and
Wang resin as a solid-phase carrier, so as to obtain a second pair of disulfide bonds; constructing an A chain sequence; s2, performing solid-phase synthesis of
insulin B chain by
Wang resin,
grafting DTDP after
cutting, and purifying; s3, butting the A chain solution and the B chain solution to form a second pair of disulfide bonds; and S4, constructing a third pair of disulfide bonds by adopting an
iodine oxidation method. According to the method, disulfide bonds can be directionally formed and constructed in three steps, non-specific crosslinking is avoided through selective oxidation of NCS, DTDP and
iodine, correct folding of molecules is ensured, the total yield of the three-step
disulfide bond forming process reaches 28% and is increased by 13% compared with one-pot oxidation, and the yield is increased; the process is controllable, and the industrial adaptability is high.