METHOD OF OBTAINING COLLAGENASE INHIBITOR WITH ANTI-TUMOUR ACTION FROM HEPATOPANCREAS OF RED KING CRAB Russian patent published in 2011 - IPC C12N9/00 

FIELD: medicine.

SUBSTANCE: method includes homogenisation of raw material in 0.1 M ammonium acetate, containing 0.1-1 mM of calcium acetate at pH 6.4-7.0. Obtained homogenate is centrifuged with obtaining supernatant. To obtained supernatant added is ammonium sulfate to 60-70% saturation. After 20 hours sediment is separated by centrifugation. Obtained sediment is dissolved in distilled water with addition of 0.1-1 mM of calcium acetate until true solution is obtained. Obtained solution is purified and concentrated on polyfiber membrane with pore size 100 kDa and washed with distilled water with further heating of concentrate to 60-100°C for 25 min with obtaining sediment. Obtained sediment is separated by centrifugation with obtaining of aggregate inhibitor preparation in 0.05 M tris buffer with pH 7.0-8.0. Obtained aggregate inhibitor preparation is chromatographed on column with affine sorbent - arginine silochrome, with further elution of active enzymes with tris buffer with addition of 1M NaCl and 20% isopropyl alcohol. Inhibitor-containing fraction is evaporated on rotor evaporator at 60-80°C for 40-60 min and concentrated by ultrafiltration on membrane with pore size 30-50 kDa obtaining concentrate. Obtained concentrate, which contains target product, is washed with 0.01 M solution of sodium chloride and freeze-dried obtaining target product.

EFFECT: extension the set of medications with anti-tumour action.

2 dwg, 5 tbl, 5 ex