BRAIN FUNCTIONS NORMALIZING AGENT AND A METHOD FOR PREPARATION THEREOF Russian patent published in 2007 - IPC A61K35/30 A61P25/00 

FIELD: pharmaceutical chemistry.

SUBSTANCE: group of inventions relate to isolation of a biologically active substances from animal brain and to preparation of therapeutical form for parenteral administration, which therapeutical form can be used in medicine as brain functions normalizing agent. Agent is prepared in therapeutical form for parenteral administration and represents peptide complex containing low-molecular weight polypeptide fraction between 70 and 90% with molecular weights of included peptides within a range of 72 to 250 Da and concentration of polypeptides 2.5-2.9 mg/mL. Such substance is obtained from brains of calves at age no more than 12 months of porcine brains via extraction with acetic acid in presence of zinc chloride. Preparation procedure involves freezing brains of calves at age no more than 12 months or porcine brains at temperature not higher than -40°C, kept at temperature from -20 to -22°C over a period not less than two months, then ground, after which 3% acetic acid is added at volume ratio 1:5 at 20±5°C. Subsequent extraction is performed at continuous stirring until homogenous slurry is formed, to which 1% zinc chloride solution is added at volume ratio 50:1. Resulting mixture is cooled at continuous stirring to 7-16°C, then stirred for 48 h at a regime of 1 h stirring followed by 4 h settling. Extract is separated from ballast materials in separator, supplemented by acetone at volume ratio 1:5, kept at 3-5°C for 4 h to form homogenized precipitate, which is re-precipitated with acetone at least 2 times. Thus obtained precipitate containing active substance is rinsed on nutsch filter with two volumes of acetone cooled to 7-16°C until light gray precipitate is obtained, which is forced through metallic riddle, dried, and dissolved in distilled water at ambient temperature and continuous stirring to concentration of polypeptides 2.5-2.9 mg/mL. Solution is centrifuged, filtered, subjected to ultrafiltration purification at back pressure no higher than 1 kg/cm² through materials with retention capacity 15000 Da. Ultrafiltrate is supplemented by glycin to its final concentration 10-20 mg/mL at pH=5.6-6.6, solution is subjected to sterilizing filtration under pressure at most 2.0 kg/cm², poured into 2-mL ampoules and autoclaved for 8 min at 120°C and atmospheric pressure 1.1 kg/cm². Thus, impurities-free aqueous solution of extract with polypeptide concentration 2.5-2.9 mg/mL is obtained. Isolated substance differs from previously obtained substances isolated from animal brains by molecular weight of peptide components. Substance is not toxic and manifests no antipyrogenic effects.

EFFECT: optimized isolation technology allowing not only preparation of impurities-free product but also increased yield thereof.

2 cl, 1 dwg, 2 tbl, 5 ex