PROTEIN INHIBITING COLLAGEN-CAUSED AGGREGATION OF MAMMALIAN PLATELETS, RECOMBINANT PROTEIN INHIBITING COLLAGEN-CAUSED AGGREGATION OF PLATELETS, DNA FRAGMENT ENCODING RECOMBINANT PROTEIN, EXPRESSION VECTOR (VARIANTS), STRAIN OF CULTURED MAMMALIAN CELLS BHK - PRODUCER OF RECOMBINANT PROTEIN (VARIANTS), METHOD OF PREPARING RECOMBINANT PROTEIN, PHARMACEUTICAL COMPOSITION Russian patent published in 1999 - IPC

FIELD: biotechnology, genetic engineering. SUBSTANCE: invention relates to protein inhibiting collagen-caused aggregation of mammalian platelets and isolated from saliva of Triatoma pallidipennis. N-Terminal amino acid sequence of this protein is determined. Protein is obtained by recombinant method and its activity is similar with that of natural protein. Amino acid sequence of recombinant protein is determined also. Also, nucleotide sequence of DNA fragment encoding this recombinant protein is determined. Expression vectors pMPSV/CMV - an inhibitor-I (DSM 7223) and pMPSV/CMV - an inhibitor-II containing DNA fragments encoding recombinant protein are constructed. Method of preparing recombinant protein is carried out by culturing mammalian cells BHK transformed with the above mentioned expression vectors that are able to produce recombinant protein. Then, protein is isolated and purified by gel-filtration on "Superose-12" and in system electrophoresis-chromatography. Pharmaceutical composition inhibiting collagen-caused aggregation of platelets has natural or recombinant protein as an active component taken at effective amount and pharmaceutically acceptable carrier or vehicle. Proteins can be used as a drug for inhibition of collagen-caused human platelet aggregation and for treatment of patients with metastatic cancer cells. EFFECT: improved method of protein preparing, enhanced antitumor effectiveness. 15 cl, 26 dwg, 1 tbl, 24 ex