In addition to the background information obtained for this patent application, NewsRx journalists also obtained the inventors' summary information for this patent application: "The objective of the claimed invention is to obtain, through a high-tech method, exopolysaccharides of bacteria S. sonnei, phase I, and develop on its basis a polysaccharide vaccines and pharmaceutical compositions.
"The technical results, provided by the claimed inventions, are: (a) obtaining native polysaccharide from S. sonnei, phase I bacteria of high purity with a high yield on a commercial scale; (b) increasing the specificity, immunogenicity, protective activity and safety of developed vaccines; © high efficacy and broad spectrum of activity of the proposed pharmaceutical compositions.
"For the first time is obtained a new polysaccharide antigen--exopolysaccharide, or capsular polysaccharide, secreted by S. sonnei, phase I bacteria into the external medium. In contrast to O-specific polysaccharide from LPS bacterial cell wall, an artificially isolated fragment of the molecule, the exopolysaccharide is an authentic natural compound, derived using S. sonnei bacteria, but without the use of LPS as its source. The primary structure of the exopolysaccharide was identical to that of the O-polysaccharide from LPS of bacteria S. sonnei, phase I, i.e. the exopolysaccharide consists of 1-100 repeating disaccharide units of O-[4-amino-2-(N-acetyl)amino-2,4- dideoxy-.beta.-D-galactopyranosyl]-(1.fwdarw.4)-O-[2-(N-acetyl)amino-2-de- oxy-.alpha.-L-altrpyranuronic acid] connected by (1.fwdarw.3) bonds to form a polysaccharide chain (FIG. 1 and FIG. 2). In contrast to the O-polysaccharide from bacterial cell LPS, the native exopolysaccharide includes a non-toxic lipid component, the composition of which contains non hydroxylated fatty acids with 16-18 carbon atoms in the molecule (FIG. 3, FIG. 4). The fatty acid content in it is no less than 0.01 (w/w) percent. Additionally, obtained by any method the exopolysaccharide from S. sonnei bacteria does not include elements of LPS core domain structure (FIG. 4). Exopolysaccharide can be prepared by any method, including genetic engineering, using the genome of S. sonnei bacteria. Preferably the exopolysaccharide is produced using S. sonnei bacteria by a method, including: (a) production of the bacterial culture in liquid phase; (b) separating the liquid phase from bacterial cells; © isolating the polysaccharide from liquid phase. Meanwhile, to avoid destroying the cell wall and LPS entry into the liquid phase, separating it from the bacterial cells is advisable to preserve the nativity of bacterial cells. Isolating the polysaccharide from the liquid phase can be carried out by a method comprising: (i) removing proteins and nucleic acids from the liquid phase; (ii) ultrafiltration and (iii) dialysis of obtained solution.
"Obtained using the above method exopolysaccharide contains no more than 1% (w/w) of protein and 2% (w/w) of nucleic acid. The molecular weight of the polysaccharide, measured by gel filtration, is from 0.4 to 400 kDa. The main fraction of the exopolysaccharide is a biopolymer with molecular weight over 80 kDa (FIG. 5B), while the main fraction of O-polysaccharide has a molecular weight of not more than 26 kDa (FIG. 5A). Exopolysaccharide is immunogenic and causes mucosal protection from shigellosis S.sonnei by inducing synthesis of a specific antibodies against S. sonnei, phase I bacteria in mammalian organisms, including humans (Example 1C, FIG. 6; Examples 2D, 2F).
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