Study On The Bioactivity Constituents Of Myristica Fragrans Houtt. And Rubus Spp.blackberry Seeds

This dissertation is composed of three chapters. The first chapter studied chemical compounds from Myristica fragrans Houtt. and bioactivity. The second chapter included chemical compounds from Rubus spp. Blackberry, and studied bioactivity. The third chapter summarized the progress of chemical constituent and pharmaceutical activity of Myristica genus.Chapter 1. The bioactivity constituents ofMyristica fragrans Houtt.The chemical constituents from M. fragrans were isolated by silica gel and Sephadex LH-20 column chromatography, and their structure were identified by NMR and MS spectroscopic data and physicochemical properties. Six compounds were obtained and identified as(7S)-8′-(benzo[3′,4′]dioxol-1′-yl)-7-hydroxypropyl)benzene-2,4-diol(1), odoratisol A(2), dehydrodiisoeugenol(3), erythro-2-(4-allyl-2,6-dimethoxyphenoxy)-1-(4-hydroxy-3-rnethoxyphenyl) propan-1-ol(4), bis(2-ethylhexyl) phthalate(5) and β-sitosterol(6). Compounds 2 and 5 were isolated from the genus for the first time.Coagulant activities of M. fragrans were assayed in vitro. PT results showed compounds 1, 4 had a certain effect on promoting exogenous coagulation, while compound 2 had a certain resistance to exogenous coagulation function; and there are certain resistance to endogenous clotting effect of all the three compounds by methods of APTT. TT revealed that three compounds promoted transfer of fibrinogen to fibrin, and FIB Showed that three compounds hd no effect on plasma fibrinolysis enzyme activity.The inhibitory activity against α-glucosidase of different elution parts separating by macroporous resin and compounds from M. fragrans were screened in a 96-well plate. In addition to the 20% elution part having inactive, other parts showed stronger inhibitory effect against a-glucosidase and were higher than that of acarbose(IC50=1136.41±150.26 μg·m L-1). The order was 40% methanol part(IC50=60.12±0.22 μg/m L) >60% methanol parts(IC50=129.27±3.47 μg/m L) > methanol part(IC50=386.80±24.37 μg/m L) > total extract(IC50=622.58±6.64 μg/m L). Moreover, compound 1 and 2 revealing strong inhibition of the α-glycosidase enzymes 1(IC50=21.76±0.59 μg/m L) and 2(IC50=21.31±0.00 μg/m L) were better than that of acarbose(IC50=1136.41±150.26 μg·m L-1), and presented a significant dose-dependence.Antibacterial activity against Staphylococcus aureus(SA), methicillin-resistant S, aureus(MRSA) and β-lactamase positive S. aureus(ESBLs-SA) of M. fragrans were assayed by the disk diffusion. Compound 1 showed the biggest inhibitory zone of 9 mm, 8 mm and 10 mm against SA, MRSA and ESBLs-SA. However, compounds 2 and 4 both only had a certain antibacterial activity to S. aureus(SA), and the inhibitory zone were 8 mm.Chapter 2. The chemical constituents of Rubus spp. BlackberryThe chemical constituents from the R. spp. Blackberry were studied by column chromatography. 13 compounds were isolated and identified as β- sitosterol(1), chrysin(2), euscaphic acid(3), hexadecanoic acid(4), 1α, 2α,3β,19α-tetrahydroxyurs-12-en-28-oic acid(5), tormentic acid(6), daucosterol(7), ursolic acid(8), oleanolic acid(9), 9(Z)-octadecenamide(10), suavissimoside R1(11), pomolic acid-28-O-β-D-glucopyranosyl ester(12), 3,5,7,2’,5’-pentahydroxy-flavan(13). Compounds 1~10, 12 and 13 were isolated from Rubus genus for the first time. Compound 11 was isolated from the plant for the first time.The result showed compound 5,7-dihydroxy flavonoid(2), euscaphic acid(3), hexadecanoic acid(4),tormentic acid(6), ursolic acid(8), oleanolic acid(9) and 9(Z)-octadecenamide(10) exhibitingsignificantly activity, and were far less than the positive control. The order was oleanolic acid(IC50=2.88±0.02 μg/m L) > ursolic acid(IC50=3.59±0.23 μg/m L) > tormentic acid(IC50=7.58±0.40μg/m L) > euscaphic acid(IC50=18.39±0.46 μg/m L) > hexadecanoic acid(IC50=58.05±1.14 μg/m L) >9(Z)-octadecenamide(IC50=66.62±0.82 μg/m L) > 5,7-dihydroxy flavonoid(IC50=85.73±3.89 μg/m L) >acarbose(IC50=1136.41±150.26 μg/m L).Chapter 3. Advances of chemical constituents andpharmacological activities of Myristica genusThe progress of chemical constituents and pharmacological research of Myristica genus were summarized.