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Pancreatitis is a global health challenge, affecting nearly 9 million people annually and resulting in more than 100,000 deaths per year. The disease, which manifests in acute and chronic forms, can be triggered and influenced by factors such as genetic factors, alcohol consumption, trauma or gallstones. Genetic studies have demonstrated the central role of trypsin in the development of pancreatitis, with risk increasing mutations in cationic trypsinogen (PRSS-1) and trypsin inhibitors (SPINK1 and PST1). The disease is initiated by the activation of trypsinogen by the lysosomal protease cathepsin b to trypsin inside the acinar cells, as shown in studies in cathepsin b knockout mice. Current treatment methods of pancreatitis, which has a mortality rate of 3-16%, focuses on a steady supply of liquids, pain management and the administration of antibiotics. Clinical trials of drugs such as BPT1 (bovine pancreatic trypsin inhibitor), camostat and gabexate mesylate (GM) have failed due to their low molecular weight and associated rapid renal clearance. The plasma half-life for BPT1 and GM is less than 1 minute, and 1-2 hours for camostat, resulting in no detectable therapeutic effect and patient death from the resulting cytokine storm. By N-terminal fusion of the Fc-region of an IgG1 antibody to the human trypsin inhibitor SPINK1 (serine protease inhibitor kazal type 1), the molecular weight of the inhibitor was increased to 64 kDa and a plasma half-life of t1/2 ≈ 21.5 hours was achieved. By Pichia pastoris strain engineering and fermentation development, a FC-SPINK1 production titer up to 40 mg･L-1 was achieved in a shaking flask fermentation under the influence of the strong methanol-inducible promoter AOX1. The fusion protein expressed in Pichia pastoris showed initial therapeutic efficacy in caerulein-induced mouse studies. This study demonstrated the prophylactic efficacy of the inhibitor in a severe mouse model of acute pancreatitis and provided insights into the immune response of the caerulein-induced mouse ... |