Study On Separation And Derivant Synthesis And Bioactives Of Polyprenols From Ginkgo Biloba L(GP)

Polyprenols from ginkgo biloba L (GP) is new effective lipids in ginkgo biloba L, whichhave similar structure as dolichols(DH) in human. GP and DH mainly take part in thebiosynthsis of glycoprotein in vivo, but GP have'nt been developed yet.The extraction and separation of GP have been studied in this dissertation. The synthesiticmechanism of polyprenyl monophosphates (GPP) was investigated from GP also. To develophealthyfood and new drugs of GP, the bioactives and pharmacology of GP and GPP forhepatoprotective properties, antitumor and antivirus were researched.The dissertation investigated systemically the analysitic methodology of GP, andestablished qualitative and quantitative scheme of TLC, RP-TLC and HPLC for GP. The contentand seasonal variation of GP were studied with C₉₅-prenol as external standard, the resultsshowed that the content of GP was lowest in spring and highest in fall, the optimum collect timeis from sept. to oct.. The yield of GP was 0.7%～0.9% with about 90% polyprenol by treatmentwith seedlings.The effection on GP with different extraction styles were systematically researched first.the ultrasonic extraction was best effective with short time, high yield and purity, itstechnological parameter was 40Khz, 150W, solid to liquid ratio (g/ml)1:10,30min,twice, theextract rate of GP was about 90% with purity 18.6%. Zauschneria extract was selected asindustrial production with over 80% yield of GP and 16%-19% polyprenols.Non-saponifialble matter of GP was made and separated by saponification and solventrefrigeration., saponifier 5%NaOH-EtOH, ratio of paste of petroleum to saponifier 1:5 (g/ml).Solvent A got GP recovery of 98.5% with 55.6% polyprenols in the twice refrigerating fluids at-10℃to -20℃, solvent B got GP revovery of 90% with 64.7% polyprenols at below -20℃inthe twice precipitates while refrigerating.Column chromatography was selected to purify non-saponifialble matter of GP. Theseperating effect was Silica gel＞alumina＞NKA-2＞polyamide＞D101. Molecular shortdistillation was first used to purify non-saponifialble matter of GP, total polyprenols could beraised from 46.2% to 83.7% in distilled remainder and there was no solvent residue, so it wouldbe the best ideal separation in industry.Sterols in lipid of GP could be separated byâ…¡grade molecular short distillation andrecrystallizatiion with a yield of 0.3-0.8% and over purity 95%. 8 sterols were identified byGC-MS, besidesβ-sitosterol, the other sterols were first reported in ginkgo biloba L.The derivants of GP were synthesized first by reaction of acylation, hydrolysis and esterifywith phosphorus oxychloride, pyro-phosphoric acid and diethyl chlorophosphate as phosphatein wild alkali, the hydrolysate was purified by silicon gel chromatography and prepared byHPLC,and the yields of monopolyprenyl phosphate and polyprenyl diethyl phosphate andpolyprenylphosphatedichlordate were over 80%, and their structure was identified with IR, ¹H-NMR,¹³C-NMR and HRMS.The bioactives and pharmacology of GP for hepatoprotective properties showed that GP isinnocuity and has a powerful protective effect on acute hepatic injury induced by carbontetrachloride, D-galactosamine and alcohol; GP and YI Shanfu could significantly decreasedserum levels of AST and ALT and significantly improved degree of proliferation of hepaticfibrous tissue and declined content of hydrop roline and collagen. GP holds significant effectson the treatment of hepatic fibrosis of chronic liver injury induced by CCl₄ in Rats.The bioactives and pharmacology of GP and GPP for antitumor showed that GPP havebetter inhibition than GP on mice transplanted tumor Heps, S₁₈₀ and EC. GP couldsignificantly prolong survival time of mice transphtnted EC tumor, nude bearing SF763 tumorand nude bearing A549 tumor. GP has good synergistic reaction on mice transplanted tumors ofHeps, S₁₈₀ and EC in combination with Fu-5, CTX, PDD and ADM, In particular the lowerdoses of GP with ⁶⁰Co radiotherapy had the best therapeutic effect on mice transplantedHeps, SoGP have distinct subsidiary effect on chemotherapy and radiotherapy and can reducetoxicity.GP had higher the clearance than CTX, and could increase remarkablely the phagocyticfunction of the macrophages, increase the index of the thymus gland in mouse with Heps andindex of the spleen in mouse with EC, The APO level in mouse with S₁₈₀ tumor in 5mg/kg GPwas 6.35, which made the rate of CD₄/CD₈ near to the normal mouse. GP have goodpharmacological functions of humoual immunity, inducing apoptosis and inhibitingproliferation.The bioactives and pharmacology of GP for antivirus in vitro showed GP is no toxicityon cell HepG 2215 and MDCK, and has better inhibition on HBV DNA secreted by cell HepG2215, with inhibition for GP 75.9% at 25μg/ml and 76.6% at 12.5μg/ml; GP had directly nokill on H₃N₂ influenza virus, but GP can raise the survival numbers of cell MDCK, the survivalrate can be raised to 55.6% at GP100μg/ml.