Study On Structure-Activity Relationship Of Anti-tumor Activity Of LAMS

Laminarin is Eeklonia kurome Okam. or Laminaria japonica Aresch. in the main polysaccharide composition, also called alginate starch.In recent years at home and abroad research shows, laminarin has antibacterial, antiviral, anti-tumor, regulate immune function, regulate blood lipids, antioxidant, anticoagulant, resistance to radiation, hypoglycemic effects, has significant value in research and development. To undertake to polysaccharide sulfated modification, in order to obtain high activity, good function of polysaccharide and its derivatives. This thesis of laminarin sulfate esterification modification, the polysaccharide and its sulfated metabolites content determination, physicochemical properties and primary structure analysis, and examines the human colon cancer LoVo cell proliferation.This paper adopts the method of chlorosulfonic acid pyridine laminarin pure (98.5%purity) of sulfated modification. Selective esterification reagent (chlorosulfonic acid and pyridine) volume ratio is1:12,1:3, reaction temperature and reaction time were75degrees C,1.5h to preparation of LAMS, LAMS-2LAMS-1.Combination of UV, IR,1H-NMR,13C-NMR, high performance liquid chromatography gel chromatography, scanning electron microscopy and other physical methods and chemical methods, preliminary study of laminarin and LAMS-1, LAMS-2physicochemical properties, content and primary structure features, the results are as follows:LAMS-2LAMS-1, the content of sulfate radical is respectively37.80%,45.92%; degree of substitution were1.07,1.51; polysaccharide content are respectively:44.52%,37.19%; molecular respectively13000,16000, the pure product of kelp polysaccharide with molecular weight of5000.The infrared spectra show, sulfated modified polysaccharide and polysaccharide compared to unmodified, about3400cm-1-OH characteristic absorption peak weakened, about810cm-1symmetric C-O-S stretching vibration characteristic absorption peak and about1245cm-1S=O asymmetric stretching vibration characteristic absorption peak appeared:1H-NMR, LAMS-1. LAMS-2hydrogen chemical displacement relative to the kelp polysaccharide chemical shift generally to low field mobile, description of kelp polysaccharide part of the hydroxyl groups have been sulfate; infrared spectrum and1H-NMR display all display laminarin and LAMS-1, LAMS-2is a beta pyranose skeleton polysaccharide.13C-NMR display LAMS-2compared to kelp polysaccharide C2and C6signals to a lower field mobile, and peak intensity is abate, C2and C6were sulfuric acid substituents; C1, C3shifi to high field:C4, C5chemical shift has no obvious change; that is LAMS-2sulfuric acid substituted position C2-OH and C6-OH; presumably the LAMS-1sulfuric acid substituted position C6-OH.Preliminary evidence of laminarin by sulfuric acid modification on sulfate radical and determine sulfate radical substitution position. Scanning electron microscopic observation of kelp polysaccharide molecules that, between the repulsive force is weaker; low degree of substitution and LAMS-1surface topography is a smooth sheet; degree of substitution of high LAMS-2surface morphology was irregular flakes and not smooth.Using the method of MTT observation of laminarin and LAMS-1, LAMS-2on human colon cancer LoVo cell proliferation inhibition, the results showed that:kelp polysaccharide and LAMS-1, LAMS-2can inhibit the growth of LoVo cells, in a dose-dependent manner; sulfated modified polysaccharide LAMS-1, LAMS-2on LoVo cell growth inhibition rate higher than the unmodified laminarin, LAMS-2on LoVo cell growth inhibition rate than LAMS-1.