Patent AU-542044-B2 (AU)
Title: Preparation of a biologically active substance for selective inhibition of the proliferation of leukemic and normal myeloid cells
Inventor: BALAZS ANDRAS
IPC: A61K35/14
Filed: 1981-01-14
Granted: 1985-02-07
Abstract: For the Contracting States : BE, CH, DE, FR, GB, IT, LI, NL, SE 1. Oligopeptide isolable from sound erythrocytes or granulocytes, respectively, selectively inhibiting the growth or proliferation, respectively, of normal and leukemic myeloid cells, characterized in that it has the gross amino acid composition Tau1 , Asx1 , Ser2 , Thr1 , Glx3 , Gly2 , Ala1 , (PO4 )1 in which Tau represents a radical of taurine, Asx means a radical of L-asparagin and/or L-asparaginic acid, Ser represents a radical of L-serine, Thr means a radical of L-threonine, Glx represents a radical of L-glutamine and/or L-glutamic acid, Gly means a radical of glycine and Ala represents a radical of L-alanine and exhibits at a pH-value of 6.5 a negative charge and a relative electrophoretic mobility of from -0.55 to -0.65 referred to asparaginic acid or at a pH-value of 1.9 no charge and a relative mobility of about 0.26 referred to epsilon-(dinitrophenyl)-lysine and has a positive reaction to chlorotolidine. For the Contracting State : AT 1. A process for preparing an oligopeptide isolable from sound erythrocytes or granulocytes, respectively, selectively inhibiting the growth or proliferation, respectively, of normal and leukemic myeloid cells having the gross amino acid composition Tau1 , Asx1 , Ser2 , Thr1 , Glx3 , Gly2 , Ala1 , (PO4 )1 in which Tau represents a radical of taurine, Asx means a radical of L-asparagin and/or L-asparaginic acid, Ser represents a radical of L-serine, Thr means a radical of L-threonine, Glx represents a radical of L-glutamine and/or L-glutamic acid, Gly means a radical of glycine and Ala represents a radical of L-alanine and exhibits at a pH-value of 6.5 a negative charge and a relative electrophoretic mobility of from -0.55 to -0.65 referred to asparaginic acid or at a pH-value of 1.9 no charge and a relative mobility of about 0.26 referred to epsilon-(dinitrophenyl)-lysine and having a positive reaction to chlorotolidine, characterized in that one a) homogenises sound erythrocytes gained from animal or human blood, particularly horse blood, in a buffer solution having a pH-value of from 7 to 8, one centrifuges the liquid homogenisate, one separates from the supernantant liquid the dissolved components having molecular weights below 10 000, one subjects this fraction to a further fractionation, suitably by gel chromatography, and separates the fraction precipitating at volume values ve /vo  of from 1.15 to 1.45 and optionally lyophilises it or b) homogenises animal organs containing granulocytes, particularly calf spleen, in an organic solvent miscible with water, one separates from the obtained liquid suspension the solids and extracts them with an organic solvent dissolving fats, one extracts the thus obtained solids with water, one separates from the liquid suspension obtained the insoluble part and one separates from the liquid phase the dissolved components having molecular weights below 10 000, one subjects this fractions to a further fractionation, suitably by gel chromatography, and one separates the fraction precipitating at volume values ve /vo  of from 1.3 to 2.5 and optionally lyophilises and one purifies the concentrate of active principle obtained according to one of the above variants a) and b) by chromatographing fractionating on account of the ionic charge and particle size and/or by paper-electrophoresis and one separates the oligopeptide having at a pH-value of 6.5 a negative charge and a relative mobility of from -0.55 to -0.65 referred to asparaginic acid or exhibiting at a pH-value of 1.9 no charge and a relative mobility of about 0.26 referred to epsilon-(dinitrophenyl)-lysine, respectively and positively reacting to chlorotolidine and optionally lyophilises it.