Synthesis And Anti-aging Evaluation Of Two Types Of Nitrogen-containing Heteroaromatic Sugar Derivatives

Pyrrolspirone ketal alkaloids and fructosazine are two types of nitrogen-containing heteroaromatic ring sugar derivatives,both of which are derived from natural products and have certain physiological and pharmacological activities.The synthesis of pollenopyrroside A derivatives and its probes was performed on the basis of total synthesis.Firstly,the reaction conditions were optimized as following:（1）The optimal conditions for the hydrodebenzylation:reaction solvent:methanol;catalyst:10%Pd/C;acidic conditions:acetic acid;（2）The optimal conditions for the dehydration of fructose to 5-HMF:catalyst:10%H₂SO₄;reaction time:3h;（3）The optimized condition of Maillard reaction between amino sugar and reducing sugar:reaction temperature:35℃;reaction time:14 h.Secondly,the3-propargyloxy-substituted probe molecule and the iodized pyrrole precursor were synthesized..In this dissertation,human embryo lung diploid fibroblasts（2BS）were cultured continuously to simulate replicative senescence.The effects of PA and 3-OH PA on cell viability were detected by MTT method.The optimal concentration of PA and 3-OH PA for delaying senescence was 0.1μmol/L.PA or 3-OH PA had obvious effects on delaying senescence and preventing senescence morphogenesis on 2BS cells at the concentration of 0.1μmol/L.The senescence-associatedβ-galactosidase（SA-β-Gal）staining method was used to evaluate the anti-aging activity.When 2BS cells were continuously cultured with 0.1μmol/L3-OH PA or PA,the expression of SAβ-galactosidase in 2BS cells was decreased mainly during the middle and late stage of cell senescence.For the 52 generation of 2BS cells,the blue staining rate was 62%in the control group,29%in the PA group,and 38%in the 3-OH PA group.The blue staining rate in the drug added culture group was significantly lower than that in the control group.Compared with 3-OH PA,PA had a stronger ability to delay aging.Different concentrations of PA had a promoting effect on the ATP content of 2BS cells.The intracellular ATP content reached the maximum when 2BS cells were cultured by 5μmol/L PA,which was 124%of the control group.The active oxygen level of 52-generation 2BS cells cultured continuously with 0.1μmol/L PA was significantly lower than that of the control group.In 2BS cells cultured at different experimental concentrations of PA or 3-OH PA after 48h,the active oxygen levels could be reduced.PA had a stronger ability to reduce active oxygen at the concentration of 10μmol/L.The IL-6 secretion level of 52-generation cells cultured with 0.1μmol/L PA for a long time was also significantly lower than that of the control group,which was 23%of that of the control group.Fructosazine was synthesized from D-gluctose by four steps,including Amadori rearrangement,isopropylidene protection,cascade reaction of hydrodebenzylation and oxidative aromatization catalyzed by Pd/C,and removal of isopropylidene protection.The structure of the key intermediate isopropylidene fructosazine was characterized by NMR,MS and X-ray single crystal diffraction.MTT assay was used to determine the effect of fructosazine on the viability of 2BS cells.The optimal culture concentration of fructosazine was 1μmol/L,which could upregulate cell viability.The SAβ-galactosidase staining method was used to evaluate the anti-aging activity of fructosazine.The blue staining rate of early-senescent and mid-senescent 2BS cells were decreased 14%and 19%,respectively,which were continuously cultured with 1μmol/L fructosazine.The results showed that fructosazine had anti-aging activity on 2BS cells in the early or middle stage of aging.