| It is very important in 21st century to find new medicine derived from ocean because many diseases are difficult to be cured at presence and vast marine resource is still not exploited. Research contents involve extraction, isolation, structural elucidation and pharmacological evaluation of metabolites from marine organism. Although many natural products are discovered every year, new medicines from them are few and existent medicine cannot satisfy present diseases and future new diseases. So it is necessary to continue the research about new medicine discovery.Our research is mainly focused on the extraction, isolation, structural elucidation and pharmacological evaluation of metabolites from marine annelid, Arenicola cristata. Some creative outcome were obtained as follows:1. Pharmacology aspects were done on the extract (ACE) from the marine annelid, Arenicola cristata including MTT assay in vitro, the antineoplastic activity in vivo by using human tumor xenograft model and the inhibition of angiogenesis demonstrated with CAM (chorioallantoic membrane) models. The results show that the extract has good antitumor bioactivity and it is very promising to develop new anticancer medicine by using Chinese translation model.2. A novel sulfate sterol, Arenicolsterol A was firstly isolated from a marine annelid Arenicola cristata and its structure was elucidated by using ESIMS, ID and 2D NMR etc. Good anticancer bioactivity of this compound was evaluated by using MTT assay and thus it is promising to be developed new medicine for common cancer therapy.3. Six anticancer compounds were obtained from the extract of marine annelid, Arenicola cristata by bioassay-guided isolation. Their primary structures were confirmed to be sterol with long saturated fatty alkyl side chain.4. Double fluorescence staining assay, DNA agarose gel electrophoresis and flow cytometry analysis were firstly been applied to study Arenicolsterol A induced Hela cells apoptosis as the deep anticancer act of this compound.5. An on-line analytical system coupling HPLC and CCD, spectrometer foridentification of the purity for each eluate was developed in a thin layerspectrometric cell. This method supplied an efficient, convenient, fast, andinexpensive approach, which would be promising in on-line identifying the purityof the samples, isolated from natural products.These outcomes and relevant research work are distributed in six chapters and now stated as followed: Chapter 1. PrefaceFirstly, research background of the paper is expatiated, which is mainly focused on potential new marine medicine discovery including anticancer, anti-HIV and anti-heart disease compounds. Secondly, the methods about isolation, structural elucidation and bioactive evaluation are stated. At last, the objects of the research work are designated to find new potential anticancer medicines from Arenicola cristata.Chapter 2. Extract and Isolation of Metabolites from the Marine Annelid, Arenicola cristata Guided by BioassayFirstly, Some information about the marine annelid, Arenicola cristata is introduced in this chapter, including species, morphologic characteristic, geographic distributing, traditional officinal part and traditional function.Secondly, apparatus, reagents and experimental methods ?for isolation are introduced. Middle or low-pressure silica gel column chromatography and sephadex LH-20 gel column chromatography are adopted; Isolation guided by bioassay is applied to discovery of the potential new medicines.Thirdly, 500 g extract were obtained from 100 kg wet Arenicola cristata after dried, minced and extracted repeatedly.At last, isolation of metabolites from the extract of Arenicola cristata guided by bioassay are expatiated, which involves partition, silica gel column chromatography, sephadex LH-20 gel column chromatography, recrystallization and HPLC. Then six pure cytotoxic compounds (ICso= 0.5 u g/ml or so) whose primary structure are sterol with long saturated fatty alkyl side chain by ID NMR were obtained. At the same time, a new sulfated sterol, Arenicolsterol A, was also obtained (yield rate is 3.2x10"6based on dry weight).Chapter 3 Arenicolsterol A, a New Anticancer Sulfated Sterol from a MarineAnnelid Arenicola cristataA new sulfated sterol, Arenicolsterol A, has been isolated from a marine annelid Arenicola cristata collected in the coast of Main land of China. The structure was elucidated using spectroscopic data including ESIMS, HNMR, CNMR, DEPT, HHCOSY, HCCOSY, HMBC, NOESY, and FTIR etc. The anticancer bioactivity of this sterol was evaluated by MTT assay. It can inhibit the growth of human cervix cancer cell line (Hela) and human non-small cell lung cancer cell line (NCI-h6) with IC50 of 3.1 ± 0.6ug/ml and 7.6 ± 0.8ug/ml.Chapter 4 Pharmacological Aspects of the Extract, Fraction and Arenicolsterol A from the Marine Annelid, Arenicola cristataCytotoxic activity of the Extract (ACE), and Fraction (ACF) from the Marine Annelid, Arenicola cristata was studied. On the basis of them, other Pharmacological aspects of ACE were investigated. The cytotoxic activity in vitro was demonstrated in two cancer cell lines (Hela cell line and SGC-7901 cell line) using MTT assay and Pharmacological aspects included the inhibition of angiogenesis demonstrated with CAM models and the antineoplastic activity in vivo by using mice tumor models. The mean IC50 ranged from 97 to 114 ng/ml of ACE and compared to the blank control, the density of the blood vessels around the sampling spots obviously became less. IR (tumour growing inhibition rate) at the dose of 0.25 g/kg.d was 80.07%.Pharmacological aspects of a novel sulfate sterol, Arenicolsterol A, isolated from marine annelid, Arenicola cristata were studied. Growth inhibition of this compound on cancer cell lines was determined by MTT assay and suppression of tumor stemcells clone forming (IC50 = 3.1 ± 0.6|xg/ml on Hela cell line, 7.6± 0.8(JLg/ml onNCI-h6 cell line). Double fluorescence staining assay, DNA agarose gel electrophoresis and flow cytometry analysis confirmed that Arenicolsterol A inhibited...
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