The debriding activity of the protease complex produced from the hepatopancreas of the king crab Paralithodes camtschatica was evaluated in vivo. The results clearly showed that the crab preparation had a high debriding ability. Individual proteases were isolated chromatographically from the crab complex to compare their debriding activities. All the purified proteases were active in vivo. The most active component of the complex was the isozyme C of crab collagenolytic protease, a member of the chymotrypsin-like protease class.

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A toxicological study of a preparation of king crab collagenase designed for local treatment of wounds shows that daily hypodermal administration of various concentrations of preparation (1.5 to 15 rng/ml) over 5 days does not affect erythrocyte morphology or hemoglobin content in laboratory animals (rats and rabbits). Systemic enzyme administration does not alter the histological structure of the kidneys (kidney is the target organ for enzyme) but leads to dose-dependent revers~le degeneration of rat liver tissues.-The milder effects of the preparation in comparison with a comparable dose of the widely used chymopsin affirm that this collagenase preparation is suitable for external use. Key words: collagenase; Icing crab; toxicology; blood components; liver; kidneysIn previous studies we have shown that a preparation of king crab (Paralithodes camtschatica) collagenase expresses high necrolytic and wound-cleaning activity [1]. The most active components of this preparation are the collagenolytic proteases A and C[11], originally described by Sakharov et al. [7].Crab collagenase accomplishes its cleaning activity by hydrolytic splitting of the fibrin coveting the wound bottom and of the collagen strands, which hold necrotic tissue on the surface of healthy tissue [3,8]. Later, in an investigation of the pharmacokinetic properties of collagenase, it was discovered that crab proteases can penetrate into the organism through the wound surface [6]. A study of the distribution of collagenolytic proteases in organs after intravenous injection reveals isoenzyme accumulation in rat liver and kidneys, whereas the protease was not detected in other organs (heart, spleen, lungs, muscles) [6]. The data obtained form a basis for a toxicological investigation of crab col/agenase. MATERIALS AND METHODSThe preparation known as "crab collagenase" with a specific activity of 600 units/mg protein (by Mandl [10]) was isolated from the hepatopancreas of P. camtschatica.Acute collagenase toxicity (LDs0) was studied on male white rats weighing 180-220 g by intravenous injection of the preparation in increasing doses. Further toxicological study was carried out on male rabbits of 2.5 kg weight and male white rats of 180-220 g weight by daily hypodermal injection during 5 days of 1 ml collagenase solution (total 6 injections) in the back. The concentrations were as follows: 15 mg/m/ (77 rats), 5 mg/ml (21 rats), and 1.5 rag/ml (21 rats). The control group (56 rats) was injected with 4 mg/ml chymopsin and benzoylarginine-p-nitroanilide (BAPNA), the activity of which was equal to the activity of collagenase in a concentration of 15 mg/ml. Seven rats received no preparation. For a comparative study of the action of the enzyme on animals of another species and weight, rabbits were injected with analogous protease doses, calculated per kg of body weight.The erythrocyte number in the peripheral blood was estimated every day. Hemoglobin concentration was recorded using spectrophotometry 0007-4888/95/0004-0377512.50 o1995 Plenum Publishing Corp...