Macrophage cytotoxin compositions and methods of preparation

Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Carbohydrate doai

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514 8, 424 851, 530395, A61K 31715

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active

051984288

ABSTRACT:
This invention provides a cytotoxin composition that inhibits mitochondrial respiration and causes cytostasis in cells, independent of nitric oxide. The invention comprises a conditioned supernatant derived from macrophage cell line (EA13.5) that inhibits mitochondrial respiration and causes cytostasis in cells. More particularly, the invention comprises a macrophage cytotoxin obtained from conditioned supernatant collected from cultured EA13.5 cell. This macrophage cytotoxin (1) inhibits mitochondrial respiration, (2) causes cytostasis independent of L-arginine derived nitric oxide, (3) is a weakly acidic (pI of about 7.5 to 8.0) glycoprotein (as determined by binding to lentil lectin sepharose). The invention also discloses a method for producing an L-arginine-derived nitric oxide-independent macrophage cytotoxin by culturing EA13.5 cells, adding IFN-.gamma. and lipopolysaccharide to trigger secretion of the macrophage cytotoxin into the conditioned supernatant, collecting the conditioned supernatant, and isolating the macrophage cytotoxin from the conditioned supernatant. Further disclosed by this invention is a method for producing a nitric oxide independent macrophage cytotoxin, that inhibits mitochondrial respiration independent of L-arginine, from cultured rodent macrophages.

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