Conjugation of succinylated gelatin to soybean trypsin inhibitor

Kunitz-type soybean trypsin inhibitor (SBTI) and succinylated gelatin fragment (Suc-gel) were conjugated by using water-soluble carbodiimide, and physicochemical and biopharmaceutical properties of the conjugate (Suc-gel-SBTI) were characterized. The conjugate contains about 2.5 mol of Suc-gel with a mean molecular weight of 32,000 per mole of SBTI, based on the content of hydroxyproline. After iv injection of the radiolabeled conjugate or native SBTI into mice, it was found that native SBTI showed rapid elimination from plasma (t(1/2 alpha) = 2 min), whereas Suc-gel-SBTI was slowly (t(1/2 alpha) = 12 min) eliminated. Organ distribution of the two agents at 30 min after iv injection was different: Suc-gel-SBTI accumulated to a large extent in both the liver and the spleen, about 23.4 and 6.9%, respectively which was more than the native SBTI. Native SBTI was taken up more rapidly by the kidney (similar to 45%) and excreted into the urine (similar to 34%) before it accumulated in other organs and tissues. In addition, we evaluated the pharmacological effects of these agents in an animal model. Suc-gel-SBTI was more effective than native SBTI against trypsin-induced lethal shock in mice. This polymer-conjugated SBTI seems to exhibit better pharmacological and therapeutic properties than its native counterpart.