A new series of flavones, thioflavones, and flavanones as selective monoamine oxidase-B inhibitors
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ContentsPART IAdvances in Flavonoid Biosynthesis Research (Ⅰ)PART IIAbstract and Key words (Ⅰ)PART IIII Introduction (1)II Flavonoids and blue flower colour (1)ⅢFlavonoids and UV-B protection in plants (2)ⅣMedicinal properties of flavonoids (3)4.1 Antioxidant activity of flavonoids (3)4.2 Vascular activity of flavonoids (3)ⅤFlavonoids biosynthesis (3)PART IV References (6)PART V Acknowledgements (7)Advances in Flavonoid Biosynthesis ResearchAbstract:The role of anthocyanins and flavones in providing stable blue flower colours in the angiosperms is outlined.The contribution of leaf flavonoids to UV-B protection in plants is critically discussed. Advances in understanding the pathway of flavonoids biosynthesis are described. The biological properties of flavonoids are considered in an evaluation of the medicinal and nutritional values of these compounds.Keywords:Flavonoid; Biosynthesis; UV-B protection; blue flower colourI IntroductionAdvances in flavonoid research over recent decades have been reviewed in a series of four volumes, beginning with Harborne et al. (1975) and culminating in Harborne. Since then, reviews of new structures in the anthocyanin and flavonoid field and with the isoflavones have appeared; as well as accounts of isoprenylated flavonoids. An introduction to flavonoids has been published (Bohm, 1999). This is essentially a listing of 6467 known flavonoid structures, with formulae, references and information on biological activities.The role of flavonoids as the major red, blue, and purple pigments in plants has gained these secondary products a great deal of attention over the years.From the first description of acid and base effects on plant pigments by Robert Boyle in 1664 to the characterization of structural and regulatory genes in the late 20th century, a wealth of information has been collected on the structures, chemical activities, and biosynthesis of these compounds. But besides providing beautiful pigmentation in flowers, fruits, seeds, and leaves, flavonoids also have key roles in signaling between plants and microbes, and in UV protection. The “early” steps in the pathway are found even in the bryophytes (mosses) and it has been suggested that synthesis of flavones, flavanones, and flavonols may have evolved first to provide chemical messengers and then UV sunscreens (Stafford, 1991). Flavonoids are also a remarkably d- iverse group of secondary products with a vast array of biological functions, including apparent roles in stress protection. The flavonols may be among the most important flavonoids in this regard; they are the most ancient and widespread of the flavonoids, synthesized even in mosses and ferns, and have a wide range of potent physiological activities.Perhaps the most active area of flavonoid research at the present time is in the possible medicinal contribution that flavonoids make to human health. Recent research on the biological properties of flavonoids will therefore be a further subject of the present review. Progress continues to be made in understanding the roles of flavonoids in stress protection, as well as in defining the mechanisms that control the amounts and varieties of flavonoids that are produced in plants in response to diverse environmental cuesII Flavonoids and blue flower colourBlue flower colour is usually due to the presence in the petals of an anthocyanin based on delphinidin. However, most delphinidin glycosides are mauve in colour and the shift to the blue region usually requires the presence of a flavone copigment, and occasionally of one or more metal cations. Blue flower colour is the preferred attractant of bee pollinators, so that evolution towards blue colour is apparent in temperate floras where bee pollination is dominant. As Gottlieb (1982) has pointed out, blue flower colour is restricted to the more highly evolved angiosperm plant families. Thus, many more primitive families have floral anthocyanins based on cyanidin (in the red to magenta range) and this explains why families like the Rosaceae and genera like Rosa lack delphinidin-based blue flowers. The chemicalbasis of blue flower colour was first extensively investigated in the case of Commelina communis.The discovery that the blue pigment of Commelina communis has two metal cations, iron and magnesium, as essential components of the anthocyanin–flavone complex, suggested at the time that metal cations might be generally present in blue flowered species. This has, however, not been borne out by subsequent experimentation. In fact, the only comparable metal complex, closely similar to that of Commelina, is that of the blue cornflower, Centaurea cyanus, where the same two metals are present and where the same ratio (6:6:1:1) of anthocyanin to flavone to iron to magnesium occurs. The only other clear example of a metal ion being required for blue flower colour is the case of the blue Hydrangea macrophylla where the metal is aluminium.Finally, mention should be made of some experiments on red–purple and red flower colours in the carnation, Dianthus caryophyllus. Red–purple carnation flowers have yielded cyanidin 3,5-diglucoside-600,6000-malyl diester, the first macrocyclic anthocyanin ever to be characterised . These two pigments are particularly unstable in acidic media and can only be extracted if neutral solvents are employed. In vivo, these two rare pigments appear to be stabilised by copigmentation with associated flavones,as with the more familiar blue copigment complexes discussed above.ⅢFlavonoids and UV-B protection in plantsUltraviolet radiation is by convention divided into three bands, each with a dierent energy and with different ecological significance. Of these, UV-B (280–315 nm) is the band of lowest wavelength and highest energy. It can penetrate the ozone layer in the stratosphere and hence potentially cause damage to plant life. Resistance to UV-B may take many forms, but one type of resistance could lie in the flavonoid pigmentss. These flavonoids generally absorb in the 280–315 region and thus are capable of acting as UV filters, thereby protecting the underlying photosynthetic tissues from damage.A series of experiments in dierent laboratories around the world have provided more convincing evidence that plants subjected artificially to UV-B radiation respond by changes in the pathway of flavonoid synthesis. Changes have been observed not only in the levels of flavonoids in epidermal cells of the adaxial leaf surface,but also in flavonoids in the leaf wax and in leaf hairs. It is interesting that Arabidopsis mutants which are blocked in the biosynthesis of related phenylpropanoids based on sinapic acid, are less aected by UV-B radiation.Although Arabidopsis is a specially favoured plant for genetic studies, mutant forms in other plants and especially in cereals can be obtained by appropriate experimentation. Another cereal in which mutants aecting flavonoid synthesis exist is barley, Hordeum vulgare.Here, a mutant has been produced which contains only 7% of the flavonoids (mainly glycoflavones based on apigenin and luteolin) of the wild type.ⅣMedicinal properties of flavonoids4.1Antioxidant activity of flavonoidsTournaireetal.(1993)havedeveloped an improved method to compare the antioxidant activity of 13 selected flavonoids from dierent classes by measuring the quantum yields of sensitised photo-oxidation of individual flavonoids. This was coupled with determination of photo-oxidation based on measuring the singlet oxygen luminescence.Previous workers (e.g. Das and Pereira, 1990) have shown that a carbonyl group at C-4 and a double bond between C-2 and C-3 are also important features for high antioxidant activity in flavonoids. Butein and other 3,4-dihydroxychalcones are more active than analogous flavones because of their ability to achieve greater electron delocalisation (Dziedzic and Hudson, 1983). In most recent reports the antioxidant activity has been measured using a lipid peroxidation assay. Rios et al. (1992) found that hypolaetin 8-glucoside (8-hydro-xyluteolin 8-glucoside) was the most potent inhibitor of non-enzymic lipid peroxidation amongst the flavone glycosides in the aerial parts of Sideritish javalambrensis(Labiatae).4.2 Vascular activity of flavonoidsIn a survey of 65 flavonoids for procoagulant activity 18 were found to inhibit the interleukin 1-induced expression of tissue factor on human monocytes but the most active was the biflavonoid, hinokiflavone (Lale et al., 1996). Tissue factor is a glycoprotein that initiates blood coagulation but this activity is not normally expressed in monocytes and endothelial cells unless they are exposed to inflammatory mediators which cause them to acquire procoagulant properties. Hinokiflavone was found to inhibit endoxin- and interleukin-induced tissue factor expression within the same concentration range with IC50 values of 18 and 48 nM, respectively.Flavonols such as kaempferol, quercetin and myricetin have been shown to inhibit adenosine deaminase activity in the endothelial cells of the aorta while flavones were found to be inactive (Melzig, 1996). Flavonoids have been shown to be potent inhibitors of the oxidative modification of low density lipoproteins by macrophages (Whalley et al., 1990). In atherosclerotic lesions lipid-laden macrophages are a characteristic feature. In any case the inhibitory eects of flavonoids in the circulation would predominate over any modification of LDL by myricetin or gossypetin.ⅤFlavonoids biosynthesisIn recent years, much effort has been directed at elucidating the flavonoid biosynthetic pathway from a molecular genetic point of view. Mutants affecting flavonoid synthesis have been isolated in a variety of plant species based on alterations in flower and seed pigmentation. Maize, snapdragon (Antirrhinum majus), and petunia were established as the first major experimental models in this system, and work in these species led to the isolation of many flavonoid structural and regulatory genes (for review, see Holton and Cornish, 1995; Mol et al., 1998). Arabidopsis more recently has helped facilitate analysis of the regulation and subcellular organization of the flavonoid pathway. One unique aspect of using Arabidopsis for studying flavonoid biosynthesis is that all but one of the enzymes of central flavonoid metabolism (leading to flavonols and anthocyanins) are encoded by single-copy genes. The exception is flavonol synthase (FLS), which appears to be encoded by six genes, two of which may not be expressed (A.Bandara, D. Owens, and B. Winkel-Shirley, unpublished data).The question of the intracellular localization and organization of flavonoid enzymes was first raised by Helen Stafford more than 25 years ago, together with the suggestion that the enzymes of general phenylpropanoid,sinapate,lignin,and flavonoid biosynthesis were likely to function as multienzyme complexes (Stafford, 1974). The concept that these pathways may be organized as enzyme complexes that facilitate the direct transfer, or channeling, of intermediates between active sites is compelling for a number of reasons. For example, there is competition for substrates at the numerous branch points within these pathways, the intermediates are highly reactive and potentially toxic, and the overall concentrations of these compounds appears to be extremely low.Many, although not all, of the enzymes of flavonoid biosynthesis are encoded by small gene families. The functional significance of this redundancy has been the subject of substantial interest over the years. One of these genes is induced by elicitor treatment, and is therefore proposed to function in the stress response related to wounding. A third enzyme can use only 6′hydroxychalcone and presumably functions in the general flavonoid pathway. It will be interesting to discover whether other legumes use similarly duplicated genes to provide for the stress-inducible synthesis of isoflavonoids.In non-legume plants, which do not synthesize isoflavonoid phytoalexins, CHI generally appears to be encoded by a single gene; although petunia does contain a second gene with homology to CHI, the product does not appear to be involved in the flavonoid pathway. Work is also underway to examine duplication in other genes of flavonoid biosynthesis, such as flavonol synthase (FLS), the only flavonoid enzyme in Arabidopsis that is encoded by a gene family. In addition to providing new informationabout the evolution of flavonoid synthesis, as discussed above, analyses of the structures of CHS and the related 2-pyrone synthase (Jeet al., 2000a) are providing a great deal of information regarding the reaction mechanisms of plant polyketide synthesis. This is opening possibilities for engineering these enzymes to produce new products similar to what has been done with the bacteria polyketide synthases. CHI, on the other hand, has a three dimensional structure and enzyme activity that are unique to plants. The structure also suggest residues that may be important in determining the different substrate specificities of CHI enzymes in legumes and other plant species. Efforts are under way to solve the structures of Arabidopsis flavonoid enzymes to facilitate the design of experiments the define the interaction interfaces of these proteins in the apparent enzyme complex.The accumulation of anthocyanin pigments in vegetative tissues is a hallmark of plant stress, yet the role that flavonoids play in the stress response is still poorly understood. In many cases, these compounds may provide antioxidant activity as part of a general stress response, which may also explain their health-promoting qualities in animals.The flavonoid biosynthetic pathway has been one of the most intensively studied metabolic systems in plants. As with any good model, each new piece of information appears to raise a number of unanticipated and intriguing questions. At the same time,new tools are providing the opportunity to consider flavonoid biosynthesis, not as an assemblage of independent components, but as part of a large, complex, and tightly orchestrated metabolic network. The ability to now consider flavonoid enzymes, for the very first time, in three dimensions and to examine the interdependence of the pathways of secondary metabolism using genomic, proteomic, and metabolic profiling methods are likely to move us much more rapidly toward this end. The development of genomics datasets for organisms that include Arabidopsis.References[1]Winkel-Shirley B: Flavonoid biosynthesis: a colorful model for genetics, biochemistry, cell biology and biotechnology. Plant Physiol 2001, 126:485-493.[2] Jeffrey B. Harborne, Christine: Advances in flavonoid research since 1986.Phytochemistry 2000,55: 481-504.[3] Antus, S., Gabor, M., Vetschera, K. (Eds.), 1995. Flavonoids and Bioflavonoids.Akademiai Kiado, Budapest.[4] Brenda Winkel-Shirley, Biosynthesis of flavonoids and effects of stress. Plant Biology2002, 5:218–222.AcknowledgementsThe author gratefully acknowledges the tutor Yang Li-min for his support and the members of her aboratory, Wang Yue and Wang Shuo, and two anonymous reviewers for helpful comments on the manuscript. Many thanks are also extended to Dong Xu-hui, Ren Yu-zhuo, and Zhang Zhi-wen for sharing inormation.8。
Strengthening basic researches on Chinese Medicinal Plants and its relations to realizing the modernization of CMM加强中国药用植物基础研究及其与中药现代化的关系Traditional Chinese Medicine (TCM) 中药Chinese traditional medicinal herbs 中草药the modernization of Chinese Materia Medica (CMM) 中药现代化Ethnobotany 人类植物学Ethnopharmacology民族药理学Ethno- 民族,种族semi-colonial and semi-feudal nation/半殖民半封建的国家multidisciplinary:多学科special projects专项调查pharmacological experiments 药理实验clinical applications临床适应性monographs 各论、专论therapeutical efficacy疗效黄花蒿( Artemisia annua),青蒿( Artemisia apiacea) chloroquine resistant malaria/抗氯喹宁疟疾Pernicious(有害的)malaria/恶性疟疾cerebral (大脑的,脑的)malaria/脑疟疾harringtonine 三尖杉酯碱homoharringtonine 高三尖杉酯碱extracted from 提取leukemia 白血病malignant lymphoma 恶性淋巴瘤ginkgetin 银杏黄酮ommon-used Chinese materia medica常用中药材品种整理和质量研究traditional Chinese patent medicines and preparations 中成药和中药制剂marketing 流通①Identification of species / 品种鉴定益胃生津,滋阴清热benefit the stomach, promote the production of body fluid and remove the excessive heat②Quality control and evaluation/质量的控制和评价intensive 加强的,透彻的remedy: n. 药物,治疗法,赔偿,modern scientific methodologies 现代科学方法①Strengthen the study of medicinal plant resources 加强药用植物基源研究re-producting resources 可持续性利用资源conservation of germplasm of the rare and endangered medicinal plants 珍稀濒危药用植物的种质资源保护②Carry out the researches on specific biology of medicinal plants③Map out GAP (Good Agriculturing Practice) in medicinal cultivation processing rules and regulations 加工方法及规范④Established standards of quality control and renew methodology⑤Apply modern comprehensive multidisciplinary studies on Chinese medicinal plants ⑥Establish information systems in modern research of Chinese medicinal herbs constraint and devastation/ 压迫与毁坏the inheritance and development of Ch inese troditional medicine ……/继承和发扬何首乌A perennial twining herb. The end of the root in a irregular plump mass.Stem somewhat woody at the base and much branched at the upperpart,herbaceous.Leave alternate,with a long periole;blade cordate,4-9cm long.2.5-5cm broad ,entire,glabrous;Flowers numerous,in terminal or axillary panicles,small;perianth 5-parted,white,the parts unequal in size.Achene elliptical,3-ridged,covered by the persisting winged perianth.多年生缠绕草本;根末端肥大呈不整齐块状,红褐色。
甘草属Glycyrrhiza L.植物资源化学及利用研究进展郑云枫;魏娟花;冷康;陶伟伟;方诗琦;彭国平;段金廒【摘要】甘草作为一种优良的天然植物资源,已被广泛应用于医药、食品、化工、畜牧等领域,国内外相关研究也方兴正艾,但近年来随着生态环境的破坏及无节制的采挖,资源状况日益严峻.本文从中药资源化学的角度出发,对甘草属植物的种类及资源分布进行总结,并针对不同品种甘草的活性成分、药理作用以及应用现状进行综述.旨在为进一步深入研究和开发该类药用植物资源提供参考,并为多层次、精细化的甘草资源综合利用提供依据.【期刊名称】《中国现代中药》【年(卷),期】2015(017)010【总页数】13页(P1096-1108)【关键词】甘草属;资源化学;活性成分;药理作用;综合利用【作者】郑云枫;魏娟花;冷康;陶伟伟;方诗琦;彭国平;段金廒【作者单位】南京中医药大学药学院,江苏南京210023;江苏省中药资源产业化过程协同创新中心,江苏南京210023;南京中医药大学药学院,江苏南京210023;南京中医药大学药学院,江苏南京210023;江苏省中药资源产业化过程协同创新中心,江苏南京210023;南京中医药大学药学院,江苏南京210023;南京中医药大学药学院,江苏南京210023;江苏省中药资源产业化过程协同创新中心,江苏南京210023;南京中医药大学药学院,江苏南京210023;江苏省中药资源产业化过程协同创新中心,江苏南京210023【正文语种】中文甘草为豆科(Leguminosae)蝶形花亚科(Papilionatae Taub.)的多年生草本植物,全球均有分布。
主产区为亚洲及欧洲,少数分布在美洲及非洲的热带和亚热带地区。
我国地处甘草资源中心地带,也是世界上甘草使用量及出口量最大的国家,年消耗量超过3万吨,以新疆、内蒙古、宁夏和甘肃为主产地,华北和西北各省区亦有产出。
国内甘草属植物约有8种[1],其中作为法定的药用品种有乌拉尔甘草Glycyrrhiza uralensis、光果甘草Glycyrrhiza glabra和胀果甘草Glycyrrhiza inflate[2]。
产品介绍列表Product C atalog1、SOY ISOFLAVONES大豆异黄酮2、Isoflavoues Aglycone大豆甙元大豆苷元3、Genistein染料木素4、Camptothecin喜树碱5、10-Hydroxy Camptothecin,10-羟基喜树碱6、7-ethyl-10-hydroxycamptothecin7-乙基-10-羟基喜树碱7、Topotecan Hydrochloride盐酸拓扑替康8、Irinotecan Hydrochloride盐酸伊立替康9、Puerarin葛根素10、Trans-Resveratrol白藜芦醇11、EGCG表没食子儿茶素没食子酸酯12、Docetaxel多烯紫杉醇13、Flaxseed Hull Extract亚麻木酚素14、Andrographolide穿心莲内酯15、Ginkgo Biloba Extract银杏叶提取物SOY ISOFLAVONES----------------------------------------------------------------------------------------------------------------------------------------Blog:/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:Product Name :SOY ISOFLAVONES 大豆异黄酮Different N ame :SIF Botanical S ource:SoybeanPrincipal C onstituent :Daidzin ,Glycerin ,Genistin ,Daidzein ,Glycitein ,Genistein.C as No.:486-66-8HS C ode :29389000.90Quality Standard :Enterprise Standard Appearance :Yellow or Light yellow Powder.Content or S PE .:5%,10%,20%,40%,60%,80%by HPLC Physics and C hemical P roperty :Is flavones is one compound,three type,sayDaidzin groups,Genistin groups and Glycitin groups.----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:Pharmacological A ction &Clinical P ractice :Fight tumour,prevent bone loose,reduce cholesterin,protect heart and relaxed women’s Menopause Syndrome.Package :25kgs/drum,N.W.:25kgs,G.W.:28kgs Dimensions:380x 520mmStorage :Stored in dry and cool place,keep away from light and 20℃temperature.HPLC A tlas tlas::Daidzein----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:Product Name :Isoflavoues Aglycone 大豆甙元,大豆苷元Different N ame :DaidzeinBotanical S ource :Trifolium pratense L.Entire gras s&Pueraria lobata(Willd.)Ohwi Root.&P.thunbergiana Benth.Root.&Medicago sativa L.Entire gras s &Glycine max (L.)Merr.SeedChemicalName :4,7dihydroxyisoflavoneCAS NO :486-66-8HS Code:29389000.90MF&MV :C 15H 10O 4and 254.23Melting Point Point::315℃~323℃Quality Standard :Enterprise Standard Appearance :White powder or Light yellow Powder.Content or S PE .:98%by HPLCPharmacological A ction &Clinical Practice Practice::Fight tumour,prevent bone loose,reduce cholesterin,protect heart and relaxed women’s Menopause Syndrome.Package :25kgs/drum,N.W.:25kgs,G.W.:28kgs Dimensions:380x 520mmStorage :Stored in dry and cool place,keep away from light and 20℃temperature.HPLC A tlas tlas::G enistein----------------------------------------------------------------------------------------------------------------------------------------Blog:/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:Product Name :Genistein 染料木素Different N ame :Genistein 染料木苷元,Botanical S ource :Trifolium pratense L.Entire gras s&Pueraria lobata(Willd.)Ohwi Root.&P.thunbergiana Benth.Root.&Medicago sativa L.Entire gras s &Glycine max (L.)Merr.SeedChemicalName :5,,7,4'-triatomicisoflavoneCAS NO :446-72-0HS Code:29389000.90MF&MV :C 15H 10O 5and 270.23Melting Point Point::297℃-298℃Quality Standard :Enterprise StandardAppearance :Yellow Powder or Light Yellow Powder.Content or S PE .:98%by HPLCPharmacological A ction &Clinical Practice :1.Have estrogenic and antioxidant activities.2.Have anti-tumor and reduce incidences of prostate cancers.3.Have anti-atherogenic and anti-osteoporotic activities.----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:4.Resist osteoporosis by preventing bone resorption and promoting bone densit.Package:25kgs/drum,N.W.:25kgs,G.W.:28kgs Dimensions:380x 520mmStorage:Stored in dry and cool place,keep away from light and 20℃temperature.HPLC A tlas tlas::----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:CamptothecinProduct Name :Camptothecin ,喜树碱Different N ame :CPTOrigin :Camptotheca acuminata DecneChemicalName :1H-pyrano[3’,4’:6,7]indolizino[1,2-6]quinoline-3,14(4H,12H)-dione,4-ethy1-4-hydroxy-(s)-CAS NO :7689-03-4HS Code:29399990MF&MV :C 20H 16N 2O 4and 348.34Quality Standard :Enterprise Standard Appearance :Yellow powder or the faint yellow Content or S PE .:98%-99%by HPLCPhysics and C hemical P roperty :Solubility264℃~267℃,[α]D25+31.3°.Meets the light to be easy to deteriorate,micro has directs wet,hasthe dextrorotation in chloroform 8with the methyl alcohol 2solutionsin.Dissolves in the lye,in ethyl alcohol,chloroform and so on inorganic solvent slightly soluble,does not dissolve in the water.Pharmacological A ction ction::The clinical experiment proved that,CPT to stomach cancer,----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:livercancer,urinary bladder cancer and leukemia and so on the malignanttumor has a better curative effect,but it has seriously not goodresponded,for example:Marrow suppression,hemorrhage cystitis anddiarrhea and so on,therefore generally not directly makes thepreparation,but takes it as the medicine intermediate to grow OPT,TPT,CPT-11and so on.Clinical Practice :This is synthesizes OPT,TPT,CPT-11and so on the intermediate Package :100g,500g,1000g/drum.Storage :Stored in dry and cool place,keep away from light and Low temperature.HPLC A tlas tlas::----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:10-Hydroxy CamptothecinProduct Name :10-Hydroxy Camptothecin ,10-羟基喜树碱Different N ame :HPT or OPT Origin :CamptothecinChemicalName :(4S)-4-Ethyl-4,9-dihydroxy-4,12-dihydro-1H-pyrano[3,4-f]quinolino-[2,3-a]indolizine-3,14-dioneCAS NO :19685-09-7HS Code:29399990MF&MV :C 20H 16N 2O 5and 364.34Quality Standard :CP2000Appearance :Straw yellow crystal powder Content or S PE .:99.5%by HPLCPhysics and C hemical P roperty :Does not dissolve to the water,slightly soluble to organic solvent,has the blue color fluorescence,the sodium salts to the water.Pharmacological A ction ction::This is another kind of alkaloid which withdraws from the camptothecaacuminata,for camptotheca acuminata alkali hydroxyl derivative,withcamptotheca acuminata alkali same,is mainly sensitive to themultiplication cell,for cell cycle specificitymedicine.Clinical Practice:This is the anti-tumor medicine raw material.Package:50g,100g,500g,1000g/drum.Storage:Stored in dry and cool place,keep away from light and Low temperature.HPLC Atlas:----------------------------------------------------------------------------------------------------------------------------------------Blog:/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:7-ethyl-10-hydroxycamptothecinProduct Name:7-ethyl-10-hydroxycamptothecin7-乙基-10-羟基喜树碱Different N ame:SN38Origin:CamptothecinChemicalName:4S)-4-Ethyl-4,9-dihydroxy-4,12-dihydro-1H-pyrano[3,4-f]quinolino-[2,3-a]in dolizine-3,14-dioneCAS NO:86639-52-3MF&MV:C22H20N2O5and392.31Quality Standard:Enterprise StandardAppearance:Faint yellow crystalline powder.Content or S PE.:99.5%by HPLCPhysics and C hemical P roperty:Light yellow needle crystal,melting point:228°C~235°C,it is easy to deteriorate when meeting light.It slightly dissolves in methanol, alcohol and chloroform.It easily dissolves in the solution of chloroform and methanol and hardly in water.It can not form stable salt with acid.Melting Point:228~235℃Clinical P ractice:Camptothecin derivative Irinotecan and SN-38glycuronic acids ----------------------------------------------------------------------------------------------------------------------------------------Blog:/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:synthesis rawmaterials.Package:50g,100g,500g,1000g/drum.Storage:Stored in dry and cool place,keep away from light and Low temperature.HPLC Atlas:----------------------------------------------------------------------------------------------------------------------------------------Blog:/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:T opotecan HydrochlorideProduct Name :Topotecan Hydrochloride 盐酸拓扑替康Different N ame :TPTOrigin :CamptothecinChemicalName :(S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-(H-Pyrano[3',4':6,7]i ndolizino[1,2-b]quinoline-3,14[4H,12H]-dionemonohydrochloride.CAS NO :119413-54-6HS Code:29399990MF&MV :C 23H 23N 3O 5.HCl ;C 23H 24ClN 3O 5and 457.91Quality Standard :Enterprise StandardAppearance :Yellow Crystal Powder.Content or S PE .:99.7%by HPLCPhysics and C hemical P roperty :Soluble in water and carbinol ;soluble a little in chloroform and ethanol soluble hardly in Acetone aether.Melting Point :213~218℃----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:Pharmacological A ction ction::Topotecan can resist lung colon rectum ovary stomach lymph galactophore cancer and leucocythemia in different degree ,especially,the curative effect of cellule lung and colon cancer is outstanding.Clinical P ractice :This is the anti-tumor medicine raw material.Package :50g,100g,500g,1000g/drum.Storage :Stored in dry and cool place,keep away from light and Low temperature.HPLC Atlas:----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:Irinotecan HydrochlorideProduct Name :Irinotecan Hydrochloride 盐酸伊立替康Different N ame :CPT—11盐酸依立替康,盐酸依列替康Origin :CamptothecinChemical Name :(+)-(4S)-4,11-Diethy[-4-hydroxy-9-[(4-piperidino-piperidino)-earbonyloxy J -1H-pyrano [3,4:6,7]indolizino [1,2-b]quinoline-3,14-(4H ,12H)-dionehydrochloridetrihydrat.CAS NO :136572-09-3HS Code:29399990MF&MV :C 33H 38N 4O 6·HCL·3H 2O and 677.19Quality Standard :Enterprise StandardAppearance :Straw yellow crystal powderContent or S PE .:99.7%by HPLCPhysics and C hemical P roperty :Soluble in water and carbinol ;soluble a little in chloroform and ethanol soluble hardly in Acetone aether.Specific R otation :+63℃-+69℃----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:/group/Herbs_Plants?hl=zh-CNPharmacological A ction ction::Irinotecan can resist lung 、colon 、rectum 、ovary 、stomach 、lymph 、galactophore cancer and leucocythemia in different degree ,especially,the curative effect of cellule lung and colon cancer is outstanding.Clinical Practice :This is the anti-tumor medicine raw material.Package :50g,100g,500g,1000g/drum.Storage :Stored in dry and cool place,keep away from light and Low temperature.HPLC A tlas tlas::----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:PuerarinProduct Name :Puerarin 葛根素Different N ame :TapoconBotanical S ource :Pueraria lobata(Willd.)Ohwi Root.or P.thunbergiana Benth.Root.ChemicalName :4H-1-Benzopyran-4-one,8-B-D-glueopyranosyl-7-hydroxy 3-(4-hydroxyphenyl)CAS NO :3681-99-0HS Code:29389000.90MF&MV :C 21H 20O 9and 416.37Melting Point Point::187℃Quality Standard :CP2005Appearance :White acicular crystalline powderContent or S PE .:99.5%by HPLCPharmacological A ction &Clinical Practice Practice::Puerarin ß-receptor interdiction ,Anti-arrhythmia,Ease angina ,Vasodilatic .Expanding brain blood vessel,anti-vas convulsion,enhancing blood stream,improving blood supplying.Reducing cholesterin,reforming platelet function,restraining its collecting and depressing its viscosity.----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:1、Vas expanding drug.Distending the vas of coronary artery and brain,falling oxygen cardiac muscle consumed ,reforming microcirculation,curing micro-blood vessel pathological changes from agedness diabetes,coronary heart disease and brain infarct.2、Treating unstable angina.3、Being effective to cure sudden deaf of all age group.4、Lowering blood pressure together with western medicine.Package:25kgs/drum,N.W.:25kgs,G.W.:28kgsDimensions:380x 520mmStorage :Stored in dry and cool place,keep away from light and 20℃temperature.HPLC A tlas tlas::----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:ResveratrolProduct Name :Trans-Resveratrol 白藜芦醇Different N ame :PolydateinBotanical S ource:Polygonum cuspidatum sieb.RootChemical Name :3',4',5-TrihydroxystlbeneCAS NO :501-36-0HS Code:29072990.90MF&MV :C 14H 12O 3and 228.25Quality Standard :Enterprise StandardAppearance :White powder.Content or S PE .:99.7%by HPLCPhysics and C hemical P roperty :Without the flavor,completely dissolves to the ethyl alcohol.Melting Point :256℃-257℃Pharmacological A ction ction::It has anti-aging function,and it is antibiotic.Antivirus,antitumor,resists the hepatitis,suppresses the agglutinate production.With plain B2of thrombus of blood platelet.It can prevent low density fat.Albumen from oxidize,have potential prevention and cure cardiovascular disease,Gives protection against cancer,antivirus andimmune regulating action,its.Function is shown as its anti-oxidant characteristic mainly. Clinical P ractice:This is the anti-tumor medicine raw material.Package:25kgs/drum,N.W.:25kgs,G.W.:28kgsDimensions:380x520mmStorage:Store in cool&dry place,Keep away from strong light and heat.HPLC Atlas:----------------------------------------------------------------------------------------------------------------------------------------Blog:/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:(-)-Epigallocatechin G allate (EGCG EGCG))Product Name :EGCG (表没食子儿茶素没食子酸酯)Different N ame :Botanical S ource :Green Tea ExtractsChemicalName :(2R,3R)-2-(3,4,5-Trihydroxyphenyl)-3,4-dihydro-1(2H)-benzopyra n-3,5,7-triol 3-(3,4,5-trihydroxybenzoate)CAS NO :989-51-5HS Code:MF&MV :C 22H 18O 11and 458.38Melting Point Point::218℃Quality Standard :Enterprise StandardAppearance :white crystal powderContent or S PE .:98%by HPLCMoisture :<5.0%Arsenide :<1ppm Heavy Metal :<10ppmSingle Impurity :<1.0%by HPLCPharmacological A ction &Clinical Practice Practice::EGCG is one kind of ingredient which withdraws from the Chinese greentea,it is green tea main activeness and the water-soluble ingredient,is in the son caffeine the content highest component,because has----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:thespecial stereochemistry structure,EGCG has the extremely strong anti-oxidized activeness,its anti-oxidized activeness at least is vitaminC 100multiple,is vitamin E 25times,can protect the cell and DNAexempts is harmed,took on the important role at the anti-cancer andthe cardiovascular disease aspect.The research indicated that,EGCGhas the remarkable anti-oxidation,eliminates in vivo free radical,the anti-tumor,the anti-sudden change,anti-is senile,the anti-inflammation,the anti-virus and so on a series of biologiesactiveness,to prevents with the treatment malignant tumor,the highblood fats sickness,the arteriosclerosis,the cerebral thrombus,thediabetes,the obesity and so on have the good function.Package:1kgs/drum,N.W.:1kgs,G.W.:1.1kgsDimensions:5x 10mmStorage :Stored in dry and cool place,keep away from light and 20℃temperature.HPLC A tlas tlas::DocetaxelProduct Name:Docetaxel,多烯紫杉醇,多西他赛Different N ame:DocetaxelBotanical S ource:ChemicalName:C as No.:114977-28-5MF&MV:C43H53NO14and807.88HS C ode:Quality Standard:Enterprise StandardAppearance:Powder.Content or S PE.:99%by HPLCPhysics and C hemical P roperty:Pharmacological A ction&Clinical P ractice:----------------------------------------------------------------------------------------------------------------------------------------Blog:/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:Package :50g,100g,500g,1000g/drum.Storage :Stored in dry and cool place,keep away from light and Low temperature.HPLC A tlas tlas::----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:Flaxseed Hull LignansProduct Name :Flaxseed Lignans or Flaxseed Hull Extract 亚麻木酚素Different N ame :Flax Lignans ,Flax Lignan ,Lignans ,Lignan ,SDGBotanical S ource:FlaxseedsChemicalName :Secoisolariciresinol Diglucoside (SDG )开环异落叶松树脂酚二葡萄糖苷(甙)CAS NO :148244-82-0HS C ode :29389000.90MF&MV :C 7H 23O 15and 347Quality Standard :Enterprise StandardAppearance :Faint yellow or yellow Powder.Content or S PE .:10%、20%、40%、60%、80%by HPLCPhysics and C hemical P roperty :Pharmacological A ction ction::Flax lignans have many health benefits for both men and women.It is extensivetly used in dietary supplements,pharmaceutical products andcosmetics.----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:Research suggests that lignans may benefit the following areas.1.Breast health ;2.Menopause ;3.Prostate health ;4.Heart health ;5.Osteoporosis ;6.Antioxidant ;7.Hair loss ;8.Acne ;9.Cholesterol ;10.Blood Suga……Clinical P ractice :Mainly applies to the capsule,the sheeting,the grain breakfast andthe fast-food food chemical additive,but also has includes omega -3fatty acid,the soluble food textile fiber and the wooden phenolelement nutrition function ultra thick emulsion and the drink and soon the many kinds of products.Package :25kgs/drum,N.W.:25kgs,G.W.:28kgsDimensions:380x 520mmStorage :Stored in dry and cool place,keep away from light and Low temperature.HPLCAtlas:----------------------------------------------------------------------------------------------------------------------------------------Blog :/中药植物提取物and /Herbs_Plantsand /herb_plant Forum:AndrographolideProduct Name :Andrographolide 穿心莲内酯Different N ame :Origin :Andrographis Paniculata (burm.f )NessChemicalName :293H)-Furanone,3-[2-[decahydro-6-hydroxy-5-(hydroxy-methyl)-5,8a-dimet hyl-2-methylene-1-naphthalenyl]ethylidene]dihydro-4-hydroxy-CAS NO :5508-58-7EINECS :226-852-5HS Code:29399990MF&MV :C 20H 30O 5and 350.44Quality Standard :Enterprise StandardAppearance :Side Lozenge or Laminated Crystallization yellowContent or S PE .:98%-99%by HPLCPhysics and C hemical P roperty :Melting Point229-232°C Alpha-126°(c=1.5,HAc)。
Why Flavonoid Phytonutrients?•Research links diets abundant in flavonoid-rich fruits, vegetables, tea, and wine with reduced risks for cancer, heart disease, and other age-related degenerative diseases.•As water-soluble antioxidants, flavonoids help protect body fluids such as blood from oxidation.•Flavonoids promote health in ways distinct from the actions of other dietary factors.Why GNLD Flavonoid Complex ™?•Broad-spectrum, water-soluble nutrient supplementation.Flavonoid Complex TM contains flavonoids representative of all flavonoid classes – flavones, flavanols,flavanones, anthocyanins and catechins – as they naturally occur in human-food-chain fruits and vegetables.•Whole-food ingredients.Our exclusive blend of flavonoid-rich extracts and concentrates is derived from whole cranberries, kale, green tea (decaffeinated),beets, elderberries, raspberries, blueberries,red and black grapes, oranges, lemons, and grapefruit. New Zealand product has added vitamin C to enhance absorption. For Australian product, combine with a vitamin C product for enhanced absorption.•Laboratory-specified amounts of key flavonoids.Flavonoid Complex TM has been tested to confirm the presence of specific flavonoids for which substantial research demonstrates benefits.•Delivers ellagic acid.Closely related to the flavonoid "family," ellagic acid is an antioxidant phytonutrient found in strawberries, cranberries, grapes, raspberries,etc.•Green tea extract.Hundreds of clinical studies show green tea flavonoids - missing in many diets - may benefit health.•No artificial colours, flavours,preservatives or sweeteners.•Convenient serving.Each tablet provides the phytonutrient value of an optimal serving of flavonoid-rich fruits and vegetables - just one more way you can meet the recommended daily intake of 6 to 7 servings of fruits and vegetables every day.“Research links diets abundant in flavonoid-rich fruits, vegetables, tea and wine, with beneficialeffects on the body.”Fast Facts About Flavonoid Complex™Flavonoids are water-soluble antioxidants in fruits, vegetables, tea, and wine. Colorful pigments, they are responsible for brilliant blues, purples, and greens, as well as yellows, oranges, and reds which cannot be attributed to carotenoids. Research links them to reduced risks for cancer, heart disease,and other age-related degenerative diseases, as well as antioxidant protection of body fluids such as blood. GNLD’s Flavonoid Complex provides diverse phytonutrients from an optimal serving of fruits and vegetables, plus catechins from green tea and ellagic acid from grapes, cranberries, and other berries.Fast Facts AboutFlavonoid ComplexTMFast Facts About Flavonoid Complex™of Lipid-soluble CarotenoidsIf carotenoids are nature’s “crayons,” creating many of the red, orange, yellow, and deep green colours in fruits and vegetables, then flavonoids can be thought of as a vivid layer in the jumbo assortment! Flavonoids are plant pigments that provide the more vibrant, brilliant colours in nature, including most of the blue, purple, and emerald green hues found in flowers, leaves, fruits, and vegetables. In addition, most of the yellow, orange, and red colours that are NOT carotenoids belong to the flavonoid family. Another distinction: carotenoids are LIPID-soluble, while flavonoids are predominantly WATER-soluble.Flavonoids represent a large family of phytonutrients (plant nutrients). More than 4,000 unique flavonoids have been identified so far — and the number is growing! Because of their chemical structure, flavonoids are part of a broader family of compounds known as polyphenols.In our diet, flavonoids are found primarily in fruits and vegetables; significant amounts of flavonoids are also found in teas and wines. Citrus fruits and berries are particularly high in flavonoids. In plants, flavonoids appear to act as a natural sunscreen, protecting against ultraviolet damage. In people, flavonoids appear to interfere with the complex and multiple processes that lead to chronic diseases such as cancer, heart disease, and other degenerative diseases associated with aging.1,2,3,4 One of the primary functions of dietary flavonoids is to act as antioxidant protectors of the watery areas in and around cells and in body fluids such as blood.Fruits and VegetablesRich in FlavonoidsThe ideal dietary goal is to consume 5 to 9 servings of fruits and vegetables every day. To help you include flavonoid-rich choices, refer to this list. The richest sources appear in italics.Rich in Anthocyanins•Berries! (blueberries, cranberries, currants, raspberries, hawthorn berries, blackberries,elderberries)•Red wine•Grapes and grape juice•Beets•Red Onions•Cherries•Citrus (grapefruit, oranges, lemons)Rich in Catechins•Green Tea (by far the richest source)•Apples•Pears•Cherries•Some WinesRich in Flavonols•Onions•Kale•Parsley•French Beans•Endive•ApplesRich in Ellagic Acid•Walnuts•Red Raspberries•Cranberries•Blackberries•StrawberriesFlavonoids and Your HealthIn the past decade, world-wide population studies have firmly established that diets high in fruits and vegetables are linked with a lower risk of developing heart disease, a wide range of cancers, and other age-related diseases.1,2,3,4Enormous efforts have been launched to identify the compounds in fruits and vegetables that may help provide protection. Because they are potent antioxidants and because they represent the majority of the plant polyphenols, flavonoids were quickly singled out as promising candidates. To date, literally hundreds of published articles report the broad protective health benefits of flavonoids. Flavonoids have clearly emerged as “hero” phytonutrients, and current research focuses on their role in helping to prevent disease.Whole foods containing flavonoids have a healthy reputation that dates back several centuries or more. In fact, many beneficial effects of traditional herbal remedies can to be attributed to the flavonoid constituents of the plants. The first clinical use of flavonoids was in 1936 when a Hungarian scientist showed that extract of citrus rinds, combined with vitamin C, was effective in strengthening capillaries, the body’s smallest blood vessels. Subsequent research also established flavonoids as anti-inflammatory, anti-allergic, and anti-infectious agents.Fast Facts About Flavonoid Complex™The Flavonoid StoryThe "Flavonoid Gap"Research continues to show that greater fruit and vegetable consumption is linked to a lower risk for developing disease. At the same time, it is evident that in many parts of the world, intake of fruits and vegetables is far from recommended levels. In Australia, for instance, 1/3of Australians do not eat the recommended amount!5,6Since fruits and vegetables supply most of the flavonoids in our diet, it is clear that our intake of flavonoids is far from optimal.The GNLD Difference in a Flavonoid SupplementFruits and vegetables are important dietary components, yet they are lacking in our diet. A part of the human food chain, they have a long history of safe use. For these reasons, GNLD set out to identify those fruits and vegetables that would deliver significant amounts of the different members of the flavonoid family. The result is Flavonoid Complex TM.Flavonoids as Nature Intended - From Whole FoodsLike carotenoids, flavonoids do not appear in isolation in nature. Specific flavonoids appear in different foods; natural foods invariably contain many flavonoids in groups. It is the DIVERSITY of foods that we eat, and the DENSITY of nutrients in those foods, that contribute to the protective health benefits. G NLD's Flavonoid Complex TM provides flavonoid diversity and density from a variety of Nature's richest whole-food sources.Broad-spectrum FormulaUsing published analytical data, GNLD developed a profile of flavonoids and related compounds present in an optimal serving of flavonoid-rich fruits and vegetables including:Beets GrapefruitBlueberries KaleCranberries LemonsElderberries OrangesGreen tea RaspberriesGrapes (red and black)Flavonoid Complex TM was then formulated so each tablet would match the profile of an optimal serving of these foods. Raw materials were carefully selected to contribute representatives of all the members of the flavonoid family, including:•AnthocyaninsThese deep purple compounds are found inblack grapes, beets, red onions and berries.•CatechinsCatechins such as +catechinsepigallocatechin, epicatechin gallate,epicatechin and epigallocatechin gallate arethe main polyphenols in green tea. Fruits suchas apples also contain catechins.•Ellagic AcidClosely related to the flavonoid family, ellagicacid is abundant in berries (particularlycranberries and raspberries) and nuts.•FlavonesFlavones include apigenin and luteolin. Theyare found in citrus fruits, red grapes and greenbeans.•FlavanolsFlavanols such as quercetin, kaempherol andmyricetin are found in many fruits andvegetables such as kale, spinach, onions andapples, as well as in black tea.•FlavanonesFlavanones include hesperidin, neo-hesperidin, naringenin and naringin.Flavanones are usually found in citrus fruitssuch as grapefruit, oranges and lemons. Flavonoids and Vitamin C:The Perfect CombinationHistorically, flavonoids have been associated with vitamin C. As early as 1939, flavonoids in citrus fruits, then referred to as "vitamin P," were known to enhance vitamin C activity. Today even more is known about the interactions of vitamin C and the large family of flavonoids represented in Flavonoid Complex TM:•Flavonoids possess vitamin C–stabilising and antioxidant-dependent vitamin C-sparingactivities.•Both flavonoids and vitamin C have complementary roles in protecting thestomach and the intestine from food-bornesubstances which can cause cancer.6•Vitamin C has been shown to enhance the ability of flavonoids to inhibit tumour growth.7•Flavonoids and vitamin C occur together in plants. Vitamin C can protect flavonoids fromoxidation.•Flavonoids enhance vitamin C absorption.Vitamin C may enhance flavonoidabsorption.8In 1972, GNLD introduced Neo-Plex Concentrate, an exclusive whole-food concentrate based on the understanding that vitamin C occurred in fruits, not in isolation, but together with a family of functionally related compounds: citrus bioflavonoids. Accordingly, GNLD's unique vitamin C supplements were formulated to include these whole-citrus compounds to assure maximum nutritional benefits.The Flavonoid StoryThe Flavonoid StoryReferences[1]Hertog, M.G. et al. Dietary antioxidantflavonoids & risk of coronary heart disease: the Zutphen elderly study. The Lancet 342: 1007-1009 (1993).[2]G ryglewski, R.J. et al. On the mechanisms ofantithrombotic action of flavonoids. Biochem.Pharmacol. 36(3): 317-22 (1987)[3]Oxidants, antioxidants and lifespan.Presentation by Dr. Lester Packer at the First Conference on Oxidative Stress & Aging. March 1994.[4]Bors, W. and Saran, M. Radical scavengingflavonoid & antioxidants. Free Rad. Res.Commun. 2: 289-294 (1987).[5]Department of Community Services & Health.National Dietary Survey of adults: 1983. No 2 Foods Consumed. Australian G overnment Publishing Service: Canberra: (1987).[6]Block, G. Dietary Guidelines and the Results ofFood Consumption Surveys: Am. J. Clin. Nutr.53:.356S-357S (1991).[7]Helser, M.A., Hotchkiss, J.H., and Roe, D.A.Influence of Fruit and Vegetable Juices on the Endogenous Formation of N-nitrosopropoline and N-nitrosothiazolidine-4-carboxylic Acid in Humans on Controlled Diets. Carcinogenesis 13:2277-2280 (1992).[8]Kandaswami, C., Perkins, E. Ascorbic AcidEnhanced Antiproliferative Effect of Flavonoids on Squamous Cell Carcinoma in Vitro.Anticancer Drugs4:91-96 (1993).[9]Green Tea Catechin Extract Symposium (video).Sponsored by Chemco Industries Inc.。
查耳酮类化合物的合成及其生物活性研究进展夏雅平;崔冬梅【摘要】近年来,随着查耳酮类化合物的生物活性的发现,对它的研究越来越多。
本文以不同的底物出发概述了查耳酮类化合物的合成方法,并对其的生物活性做了一下总结。
%Chalcones have important biological effects,so they get more attentions. In this paper, we summarize some synthetic methods of the chalcones derivatives from different starting substrates, and give a introduction about it's bioactivity.【期刊名称】《浙江化工》【年(卷),期】2011(042)009【总页数】4页(P11-13,16)【关键词】查耳酮;合成;活性【作者】夏雅平;崔冬梅【作者单位】浙江工业大学药学院,浙江杭州310014;浙江工业大学药学院,浙江杭州310014【正文语种】中文【中图分类】X830.70 前言查耳酮类化合物是合成黄酮类化合物的重要中间体,在有机合成中也有着特殊的用途,因此其制备方法受到普遍关注。
随着查耳酮类化合物的生物活性的逐渐发现,对它的研究也越来越深入。
以下介绍查耳酮类化合物主要的合成方法及其生物活性。
1 查耳酮类化合物主要的合成方法1.1 以一氧化碳,苯乙烯,碘苯为原料[1]该反应由苯乙烯出发,与一氧化碳和卤代苯在100℃,5atm和钯催化剂的条件下,在苯乙烯和卤代苯之间插入一个羰基从而得到α,β-不饱和酮。
这个反应的适用性比较广,收率也比较高,但是钯配体的结构比较复杂,且反应时间要20h,不适合工业化的大规模生产。
1.2 以肉桂酰氯和三苯基铋为原料[2]该反应是3mol的肉桂酰氯和1mol三苯基铋在钯催化剂的作用下,通过交叉偶联反应,生成α,β-不饱和酮类化合物,虽然在适当的溶剂和碱的条件下收率可以达到60%以上,但是三苯基铋的价格比较昂贵,限制了该方法的使用。
黄酮类化合物抗癌作用机制的研究进展药学文献检索作业论文作者姓名: 渠文涛作者学号: 201012282785学专业: 药物化学所论文完成时间: 2010.12.302010 年 12月 30日黄酮类化合物抗癌作用机制的研究进展摘要: 黄酮类化合物抗肿瘤作用的研究由来已久,目前已发现具有抗肿瘤作用的黄酮类化合物主要有槲皮素、水飞蓟素、芦丁、柚皮甙等。
黄酮类化合物尽管在体外试验中表现出很强的抗氧化能力,但由于在人体血浆中的稳态浓度低,所以作为抗氧化剂产生的抗癌作用是较小的。
黄酮类化合物预防癌症的作用主要与其抑制硫氧还蛋白还原酶、抑制基质金属蛋白酶和抑制信号转导等作用机制有关。
关键词:黄酮类化合物; 类黄酮; 抗癌; 抗氧化;抗肿瘤Abstract : People have studied Flavonoids` anti-tumor effect for a long time, and have found lots of flavonoids has anti-tumor effect ,such as quercetin, silymarin, rutin, naringin and so on. Flavonoids, despite their strong antioxidant capacity in vitro, have low steady-state concentrations in human plasma, and thus, as antioxidants, have very little anti-cancer effec.t Their cancer preventing effect can be attributed mainly to their ability to interact with basic cellularmechanisms. Such interactions include the inhibition of thioredoxin reductase, inhibition of matrix metalloproteinases and inhibit the role of signal transduction ,and so on.Key words: Flavonoids; flavonoids; anticancer; antioxidant; antitumor黄酮类化合物(Flavonoids,又称类黄酮化合物),主要指具有色酮环与苯环为基本结构的一类化合物的总称,具C-C-C基本构型,为植物体多酚类的代636谢物.根据黄酮类化合物化合物的结构不同,可将其进一步分为黄酮醇类(Flavonols)、黄酮类(Flavones)、黄烷酮类(Flavanones)、黄烷醇类(Flavanols)[或儿茶素类(Catechins)]、花色素类(Anthocyanins)、异黄酮类(Isoflavones)、二氢黄酮醇类(Dihydroflavonols)以及查耳酮类[1-2](Chalcones)等等。
A new series of flavones,thioflavones,and flavanones as selective monoamine oxidase-B inhibitorsFranco Chimenti a ,Rossella Fioravanti a,*,Adriana Bolasco a ,Paola Chimenti a ,Daniela Secci a ,Francesca Rossi a ,Matilde Yáñez b ,Francisco Orallo b ,Francesco Ortuso c ,Stefano Alcaro c ,Roberto Cirilli d ,Rosella Ferretti d ,M.Luisa Sanna daDipartimento di Chimica e Tecnologie del Farmaco,Universitàdegli Studi di Roma ‘‘La Sapienza ”,P.le A.Moro 5,00185Roma,Italyb Departamento de Farmacología and Instituto de Farmacia Industrial,Facultad de Farmacia,Universidad de Santiago de Compostela,Campus Universitario Sur,E-15782Santiago de Compostela (La Coruña),Spain cDipartimento di Scienze Farmacobiologiche,Universitàdi Catanzaro ‘‘Magna Graecia ”,‘‘Complesso NinìBarbieri ”,88021Roccelletta di Borgia (CZ),Italy dIstituto Superiore di Sanità,Dipartimento del Farmaco,Viale Regina Elena 299,I-00161Rome,Italya r t i c l e i n f o Article history:Received 2November 2009Revised 2December 2009Accepted 8December 2009Available online 4January 2010Keywords:FlavonesMAO inhibitorsMolecular modelingEnantiomeric separationa b s t r a c tA new series of synthetic flavones,thioflavones,and flavanones has been synthesized and evaluated as potential inhibitors of monoamine oxidase isoforms (MAO-A and -B).The most active series is the flava-none one with higher selective inhibitory activity against MAO-B.Some of these flavanones (mainly the most effective)have been separated and tested as single enantiomers.In order to investigate the MAOs recognition of the most active and selective compounds,a molecular modeling study has been performed using available Protein Data Bank (PDB)structures as receptor models for docking experiments.Ó2009Elsevier Ltd.All rights reserved.1.IntroductionFlavonoids are an extensive group of polyphenolic compounds present in plants,regularly consumed foods (e.g.vegetables and fruits),olive oil,and beverages like tea and wine.1–3They are usu-ally subdivided,according to their chemical structure,into several subclasses including anthocyanidins,flavanones,flavones,flavo-nols,flavanonols (or dihydroflavonols),chalcones,isoflavones,and flavanols (flavan-3-ols)(also called catechins).Besides their physiological role in plants,these different flavonoids have been reported to possess a wide range of biological activities,including modulatory properties of several enzymes [e.g.activation of sirtu-ins,4inhibition of monoamine oxidase (MAO),5and a number of other biological activities such as anxiolytic,6anti-inflammatory,3antiviral,7antiprotozoal,8antioxidant,3cardiovascular,3,9and anti carcinogenic properties.10–12Concerning the effects of flavonoids on the enzymatic activity of MAO (EC 1.4.3.4),several studies 5,13–15have previously described the MAOIs (MAO Inhibitors)properties of jaceosidine,eupafolin,luteolin,quercetin,and apigenin (Chart 1).Further in our recent paper 16we have reported the activity of the flavone quercetin,which showed a selective MAO-A inhibitory activity in the nanomolar range (IC 50=10nM).0968-0896/$-see front matter Ó2009Elsevier Ltd.All rights reserved.doi:10.1016/j.bmc.2009.12.029*Corresponding author.Tel.:+39649693259;fax:+3964462731.E-mail address:rossella.fioravanti@uniroma1.it (R.Fioravanti).Bioorganic &Medicinal Chemistry 18(2010)1273–1279Contents lists available at ScienceDirectBioorganic &Medicinal Chemistryj o u r n a l h o m e p a g e :w w w.e l s e v i e r.c o m /l o c a t e /b m cMAO is aflavoprotein located in the outer membrane of the mitochondria that contains a covalently boundflavin adenine dinucleotide(FAD)as a coenzyme and that has considerable phys-iological and pharmacological interest due to its central role in the metabolism of monoamine neurotransmitters.MAO exists in two isoforms,MAO-A and MAO-B,which share approximately70% sequence identity on the amino acid levels and differ in their substrate specificity,susceptibility to specific inhibitors,and three-dimensional structure.17,18As indicated above,these MAO isoforms catalyze the biotransformation by oxidation of endoge-nous neurotransmitter monoamines and also the metabolization of various exogenous primary,secondary,and tertiary amines in the central nervous system as well as in peripheral tissues.19 MAO-A preferentially deaminates5-hydroxytryptamine(5-HT), norepinephrine,and epinephrine and is irreversibly inhibited by low concentrations of clorgyline whereas MAO-B preferentially deaminates dopamine,b-phenylethylamine,and benzylamine and is irreversibly inhibited by R-(À)-deprenyl.It is well known that major depression is related to the deficit of monoamines(basically norepinephrine and5-HT)at critical syn-apses in the central nervous system20whereas Parkinson’s disease is mainly due to a deficit of dopamine.21All thesefindings support the clinical importance of MAO inhib-itors in the treatment of several neurological and psychiatric disor-ders.Thus,selective MAO-A inhibitors are used as anti-depressant and anti-anxiety drugs whereas selective MAO-B inhibitors can be mainly used to treat Parkinson’s disease either alone or in combi-nation with L-DOPA.22,23Bearing in mind the above considerations,we consider that nat-ural products asflavonoids(e.g.flavones andflavanones)are promising lead compounds for developing effective agents to com-bat neurological diseases.Therefore,as part of our continuous search for potential selective MAO inhibitors,we have synthesized flavones,thioflavones,andflavanones to investigate their inhibi-tory activity on human MAO isoforms(hMAO-A and-B).Due to the presence of a stereogenic carbon on theflavanone ring(flava-nones exist as(R)-and(S)-enantiomers),we performed on the most active compounds the enantioseparation by chiral HPLC and tested the single enantiomers.Furthermore,the most promis-ing compound binding mode to both hMAO-A and B catalytic clefts has been investigated by means of molecular modeling carried out using PDB crystallographic structures as receptor models for dock-ing simulations.2.ChemistryThe syntheses of theflavones andflavanones derivatives were carried out using chalcones as starting products24,25;(Scheme1) whereas the thioflavones derivatives were prepared from the pre-viously synthesizedflavones.The synthesis of chalcone derivatives was performed according to procedures previously described.26 Briefly,we used the aldolic condensation with Ba(OH)2Á8H2O as base.In this process,20-hydroxyacetophenone reacted with an aldehyde.Chalcones1a–p and sodium acetate were heated in refluxing ethanol to obtain derivatives2a–p.To obtain derivatives 3a–p and4a–p,suitable chalcones1a–p were dissolved in DMSO (dimethyl sulfoxide)and iodine was added.After oxidative cycliza-tion of chalcones,the derivatives3a–p were treated with Lawes-son’s reagent,27in refluxing benzene for2h to obtain4a–p derivatives.3.BiochemistryThe potential effects of the test drugs on hMAO activity were investigated by measuring their effects on the production of hydrogen peroxide(H2O2)from p-tyramine(a common substrate for both hMAO-A and-B),using the Amplex Red MAO assay kit (Molecular Probes,Inc.,Eugene,Oregon,USA)and microsomal MAO isoforms prepared from insect cells(BTI-TN-5B1-4)infected with recombinant baculovirus containing cDNA inserts for hMAO-A or-B(Sigma–Aldrich Química S.A.,Alcobendas,Spain).The production of H2O2catalyzed by MAO isoforms,can be de-tected using10-acetyl-3,7-dihydroxyphenoxazine(Amplex Red re-agent),a non-fluorescent compound and highly sensitive probe that reacts with H2O2in the presence of horseradish peroxidase to produce afluorescent product,resorufin.In this study hMAO activity was evaluated using the above method following the gen-eral previously procedure described by.28The test drugs(new compounds and reference inhibitors)them-selves were unable to react directly with the Amplex Red reagent, which indicates that these drugs do not interfere with the mea-surements.On the other hand,in our experiments and under our experimental conditions,the control activity of hMAO-A and-B (using p-tyramine as a common substrate for both isoforms)was 165±2pmol of p-tyramine oxidized to p-hydroxyphenylacetalde-hyde/min(n=20).Most testedflavonoids concentration-dependently and selec-tively inhibited the enzymatic control activity of MAO-B(see Ta-bles1–3).4.Results and discussionAll synthesized compounds2a–p,3a–p,and4a–p were assayed for hMAO-A and-B inhibitory activity.By analyzing the data re-ported in Tables1–3it can be seen that the most active and hMAO-B selective compounds belong toflavanones2a–p deriva-tives and among them the best results(in the low micro molar range)were observed with compounds2f,2g,2h,2j,2k,and2n, substituted with thefluorine atom and the methyl or the methoxy group.It seems that the substitution pattern of these groups do not1274 F.Chimenti et al./Bioorg.Med.Chem.18(2010)1273–1279affect the activity.The introduction of a double bond in the flavones3a–p and of the sulfur in the thioflavones4a–p leads to a relevant decrease of the inhibitory activity although remain a hMAO-B selectivity.The most active compounds2f,2g,2h,2j, 2k,and2n,first assayed as racemates,were resolved by enantiose-lective HPLC and tested again as single enantiomers.The direct HPLC enantiomeric separation was accomplished on the immobi-lised polysaccharide-based Chiralpak IA chiral stationary phases (CSP)using pure methanol as a mobile phase.The absolute config-uration of the enantiopure forms isolated at semipreparative scale was empirically assigned by comparing the circular dichroism(CD) spectra of the enantiomers offlavanone29with those of the struc-turally analogues2f,2g,2h,2j,2k,and2n(see Supplementary data).In the reversibility and irreversibility tests,hMAO-B inhibition was irreversible in presence of the compounds selected for docking experiments[2j,(R)-2j and(S)-2j](see below)as shown by the lack enzyme activity restoration after repeated washing.Similar results were obtained for R-(À)-deprenyl(Table4).However,significant recovery of hMAO-A activity was observed after repeated washing of moclobemide,indicating that this drug is a reversible inhibitor of hMAO-A.Compound2j,that evidenced the best inhibitory activity as racemate,was the best compound also in the two enantiomeric forms(Table5).Because of this result we selected this compound for the subsequent molecular modeling investigation.The recognition of both(R)-2j and(S)-2j enantiomers was eval-uated by means of docking studies with respect to hMAO-A and-B receptor models.Two new,high resolution,PDB crystallographic structures,after a preliminary treatment,were adopted as targets (see Experimental section).The molecular modeling investigation, in qualitative agreement to the experimental data,reported the capability of both2j enantiomers tofit within hMAO-A and-B cat-alytic sites with some differences in terms of affinity.Actually both isomers recognized preferentially the hMAO-B binding cleft and, between them,the(S)-2j highlighted most favorable interactions.In order to analyze the binding modes of2j enantiomers within the MAO isoforms,the most stable theoretical complexes were graphically inspected reporting similar interaction and binding modes.Furthermore in Table6was reported the average G score values for the modeled compounds.As displayed in Figure1and according to interaction energy data,the(R)-2j showed a more productive recognition with respect to the(S)-2j into the hMAO-A.In the hMAO-A the(R)-2j was able to deeply penetrate into the active sites highlighting hydrophobic contacts to the FAD.Such an obser-vation cannot be addressed to the(S)-2j located at a larger distance from the cofactor.Taking into account the orientation of both enantiomers into the hMAO-A binding cleft,(S)-2j appeared slightly translated toward the outer side.So,the most part of inter-acting residues was shared between the enantiomers but(R)-2j performed stronger interaction to Tyr407and Tyr444with respect to(S)-2j who,vice versa,was able to hydrophobically contact Ala111,Ile325,and Leu97.A similar scenario was observed into the hMAO-B case but the binding mode differences were less remarkable(Fig.2).Although(R)-2j bound relatively deeper in the active site cavity, the(S)-enantiomer also was recognized by the FAD cofactor.The Table2Structures and inhibitory activity of derivatives3a–pORR1OCompd R R1IC50l M hMAO-A IC50l M hMAO-B SI–3a H H*****3b H F******3c H CH3** 2.38±0.07>42b 3d H OCH3***27.87±1.04>3.6b 3e F H***12.94±0.47>7.7b 3f F F*****3g F CH3** 4.33±0.39>23b 3h F OCH3*** 1.34±0.02>75b 3i CH3H****3j CH3F*****3k CH3CH3******3l CH3OCH3******3m OCH3H*****3n OCH3F** 5.14±0.18>19b 3o OCH3CH3**11.34±0.26>8.8b 3p OCH3OCH3**49.72±1.83>2b Clorgyline(IC50l M hMAO-A0.004±0.0003a;IC50l M hMAO-B61.35±1.13;SI# 0.000065);R-(À)-deprenyl(IC50l M hMAO-A67.25±1.02a;IC50l M hMAO-B 0.019±0.0009;SI# 3.539);iproniazid(IC50l M hMAO-A 6.56±0.76;IC50l M hMAO-B7.54±0.36;SI#0.87);Moclobemide(IC50l M hMAO-A361.38±19.37;IC50 l M hMAO-B*;SI#<0.36c);–SI:hMAO-B selectivity index=IC50(hMAO-A)/IC50(hMAO-B). Each IC50value is the mean±S.E.M.fromfive experiments.Level of statistical sig-nificance:a P<0.01versus the corresponding IC50values obtained against hMAO-B, as determined by ANOVA/Dunnett’s.b Values obtained under the assumption that the corresponding IC50against hMAO-A is the highest concentration tested (100l M).c Value obtained under the assumption that the corresponding IC50 against hMAO-B is the highest concentration tested(1mM).*Inactive at1mM(highest concentration tested).**Inactive at100l M(highest concentration tested).***100l M inhibits by approximately40%.Table1Structures and inhibitory activity of derivatives2a–pORR1OCompd R R1IC50l M hMAO-A IC50l M hMAO-B SI#2a H H** 2.87±0.09>35b2b H F** 4.41±0.13>23b2c H CH3** 2.07±0.11>48b2d H OCH3** 2.20±0.10>45b2e F H** 3.77±0.16>27b2f F F**0.67±0.04>149b2g F CH3**0.16±0.006>625b2h F OCH3**0.17±0.005>588b2i CH3H** 3.62±0.10>28b2j CH3F**0.13±0007>769b2k CH3CH3**0.50±0.02>200b2l CH3OCH3** 3.35±0.07>30b2m OCH3H** 1.47±0.03>68b2n OCH3F**0.17±0.009>588b2o OCH3CH3**0.55±0.02>182b2p OCH3OCH3**7.18±0.06>14bClorgyline(IC50l M hMAO-A0.004±0.0003a;IC50l M hMAO-B61.35±1.13;SI#0.000065);R-(À)-deprenyl(IC50l M hMAO-A67.25±1.02a;IC50l M hMAO-B0.019±0.0009;SI# 3.539);Iproniazid(IC50l M hMAO-A 6.56±0.76;IC50l MhMAO-B7.54±0.36;SI#0.87);Moclobemide(IC50l M hMAO-A361.38±19.37;IC50l M hMAO-B*;SI#<0.36c);–SI:hMAO-B selectivity index=IC50(hMAO-A)/IC50(hMAO-B).Each IC50value is the mean±S.E.M.fromfive experiments.Level of statistical significance:a P<0.01versus the corresponding IC50valuesobtained against hMAO-B,as determined by ANOVA/Dunnett’s.b Values obtainedunder the assumption that the corresponding IC50against hMAO-A is the highestconcentration tested(100l M).c Value obtained under the assumption that thecorresponding IC50against hMAO-B is the highest concentration tested(1mM).*Inactive at1mM(highest concentration tested).**Inactive at100l M(highest concentration tested).F.Chimenti et al./Bioorg.Med.Chem.18(2010)1273–12791275number of interacting residues was equivalent and a large part of them was shared.So the slightly advantage of (S )-2j interaction en-ergy with respect to (R )-2j could be addressed to the better accom-modation of its fluorophenyl moiety into the lipophilic cage delimited by Trp119,Phe168,Leu171,Ile199,and Tyr326.The analysis of the residues involved in the recognition of 2j enantio-mers in both MAO isoforms revealed that the selectivity was not addressed to canonical differences between the catalytic gorges such as Ile199and Tyr326(hMAO-B),respectively,versus Phe208and Ile335(hMAO-A).Actually all these crucial amino acids interacted with both 2j enantiomers as shown in Figures 1and 2.Since the average G score values (see Table 6)take into account other effects (solvation,pen-alty clashes,loss of ligand internal degrees of freedom,etc)the hMAO-B selectivity can be addressed to those terms.Finally in or-der to clarify the role of R 1we have compared the hMAO-BTable 3Structures and inhibitory activity of derivatives 4a –pORR 1Compd R R 1IC 50l M hMAO-AIC 50l M hMAO-BSI –4a H H *****4b H F ****4c H CH 3** 6.69±0.13>15b 4d H OCH 3***0.48±0.024>208b 4e F H ******4f F F ** 6.89±0.19>15b 4g F CH 3*** 1.61±0.04>62b4h F OCH 3*****4i CH 3H *****4j CH 3F *****4k CH 3CH 3** 6.72±0.08>15b4l CH 3OCH 3*****4m OCH 3H *****4n OCH 3F *****4o OCH 3CH 3***7.61±0.43>13b 4pOCH 3OCH 3**3.34±0.16>30bClorgyline (IC 50l M hMAO-A 0.004±0.0003a;IC 50l M hMAO-B 61.35±1.13;SI #0.000065);R-(À)-deprenyl (IC 50l M hMAO-A 67.25±1.02a ;IC 50l M hMAO-B 0.019±0.0009;SI # 3.539);Iproniazid (IC 50l M hMAO-A 6.56±0.76;IC 50l M hMAO-B 7.54±0.36;SI #0.87);Moclobemide (IC 50l M hMAO-A 361.38±19.37;IC 50l M hMAO-B*;SI #<0.36c )SI:hMAO-B selectivity index =IC 50(hMAO-A)/IC 50(hMAO-B).Each IC 50value is the means ±SEM from five experiments.Level of statistical significance:a P <0.01versus the corresponding IC 50valuesobtained against hMAO-B,as determined by ANOVA/Dunnett´s.b Values obtained under the assumption that the corresponding IC 50against hMAO-A is the highest concentration tested (100l M).c Value obtained under the assumption that the corresponding IC 50against hMAO-B is the highest concentration tested (1mM).*Inactive at 1mM (highest concentration tested).**Inactive at 100l M (highest concentration tested).***100l M inhibits by approximately 40%.Table 4Reversibility and irreversibility of hMAO inhibition of compound 2j and of its separated enantiomers Compd%hMAO-A inhibition%hMAO-B inhibitionBefore washingAfter repeated washing Before washing After repeated washing (R )(À)D a (20nM)51.45±2.6952.05±2.88M a (500l M)84.75±4.3410.26±0.65b2j (100nM)38.36±3.1536.51±2.56(R )(+)2j (100nM)35.61±2.9037.23±3.14(S )(À)2j (100nM)45.29±3.9242.58±3.37a D ,(R )(À)deprenyl;M ,moclobemide.Each value is the means ±SEM from five experiments (n =5).bLevel of statistical significance:P <0.01versus the corresponding %hMAO-A inhibition before washing,as determined by ANOVA/Dunnett´s.Table 5Inhibitory activity of single enantiomer of the most active flavanones 2Comp.IC 50l M hMAO-A IC 50l M hMAO-B SI a (R )(+)2f **0.65±0.03>154b (S )(-)2f **0.62±0.02>161b (R )(+)2g **0.15±0.01>667b (S )(À)2g **0.17±0.01>588b (R )(+)2h **0.15±0.01>667b (S )(À)2h **0.15±0.01>667b (R )(+)2j **0.14±0.01>714b (S )(À)2j **0.11±0.01>909b (R )(+)2k **0.53±0.03>189b (S )(À)2k **0.59±0.03>169b (R )(+)2n **0.20±0.01>500b (S )(À)2n**0.16±0.01>625bEach IC 50value is the means ±SEM from five experiments.aSI:hMAO-B selectivity index =IC 50(hMAO-A)/IC 50(hMAO-B).bValues obtained under the assumption that the corresponding IC 50against MAO-A is the highest concentration tested (100l M).Table 6Theoretical affinity of 2j ,(R )-2j ,and (S )-2j with respect to MAO isoforms Comp hMAO-A *hMAO-B *(±)-2j À6.75À7.45(R )-2j À6.93À7.44(S )-2jÀ6.58À7.47*Average G score values.Figure 1.Superimposition of best fully energy minimized hMAO-A poses of (R )-2j and (S )-2j .The ligand is reported as white and green carbon polytube,respectively.The interacting residues with (R )-2j and (S )-2j are colored in white and green,respectively.The FAD cofactor is displayed as a spacefill structure.Non-polar hydrogen atoms are omitted for clarity.1276F.Chimenti et al./Bioorg.Med.Chem.18(2010)1273–1279interaction energies of 2j ,2i ,2k ,and 2l averaging the values of both enantiomers.The results (not shown)indicated the fluoro-phenyl as the best moiety with the optimal compromise of lipo-philicity and steric hindrance.In principle,our results of irreversibility obtained with the compounds 2j ,(R )-2j ,and (S )-2j ,do not seem to correlate well with the conformational analysis made in the docking studies but a number of arguments may ex-plain,at least in part,this discrepancy.In most cases,the irrevers-ible inhibitors establish a covalent interaction with the active center of the enzyme.30However,not all irreversible inhibitors form covalent adducts with their enzyme targets but they may also act by other mecha-nisms.In fact,some reversible inhibitors bind so tightly to their target enzyme that they are essentially irreversible.These tight-binding inhibitors may show kinetics similar to covalent irrevers-ible inhibitors.In these cases,some of these inhibitors rapidly bind to the enzyme in a low-affinity enzyme-inhibitor (EI)complex and this then undergoes a slower rearrangement to a very tightly bound EI*complex.This kinetic behavior is called slow-binding.This slow rearrangement after binding often involves a conforma-tional change as the enzyme ‘‘clamps down”around the inhibitor molecule.31In contrast,some irreversible MAO inhibitors (the so-called suicide inhibitors)act as a substrate for the target enzyme,which finally generates a new compound that irreversibly inhibits MAO activity.Therefore,the initial interaction of these inhibitors with MAO may be different to the interaction obtained after sev-eral minutes of the enzyme–inhibitor complex formation (see above).R-(À)-deprenyl,for example,first of all form a non-covalent complex with MAO as an initial,reversible step.The sub-sequent interaction of R-(À)-deprenyl with MAO leads to a reduc-tion of the enzyme-bound FAD,and concomitant oxidation of the inhibitor.This oxidized inhibitor then reacts with FAD at the N-5-position in a covalent manner.32The initial non-covalent bind-ing to MAO has been also described for other MAO inhibitors (e.g.clorgyline derivatives).33Finally,it is possible that,in some cases of irreversible inhibition,a steric hindrance may prevent the release of the inhibitor from the enzymatic active center (although its interaction with this binding site is either very weak or reversible).Bearing in mind all the above considerations and taking into ac-count that the docking studies make only a theoretical prediction of the initial possible interaction inhibitor–enzyme,the results ob-tained in these docking studies and in the reversibility experiments may be different.5.Experimental5.1.General procedure for the synthesis of 20-hydroxy chalcones 1a–pA solution of suitable 2-hydroxy acetophenone (0.01mol)and the suitable benzaldehyde (0.01mol)dissolved in ethanol was treated with barium hydroxide (0.01mol).The solution was stirred 24h at 30°C.After the pH of the reaction mixture was brought back to 7.0by the careful addition of HCl 1N solution.The aqueous layer was extracted with ethyl acetate,washed with brine,dried (Na 2SO 4),and then concentrated in vacuo.The residue was purified from ethanol,to afford chalcones 1a–p .5.2.General procedure for the synthesis of flavanones 2a–pChalcones 1a–p (0.2mmol)and sodium acetate (2.0mmol)were heated in refluxing ethanol (2mL)for 24–48h.The mixture was then to cool to r.t.and poured into ice water (10mL)and extracted with CH 2Cl 2(3Â10mL).The combined or-ganic phase was washed with brine,dried over Na 2SO 4,and then concentrated in vacuo.The residue was column chromatographed on silica gel,eluting with mixture of ethyl acetate/hexane 1:4,to afford flavanones 2a–p which were crystallized from suitable sol-vent (see Supplementary data).5.3.General procedure for the synthesis of flavones 3a–p To a solution of chalcones 1a–p (0.015moL)in 50ml of DMSO (dimethyl sulfoxide)were added iodine (0.015mol).The reaction mixture was stirred for 30min at 130°C.Then the mixture was treated with sodium thiosulfate (solution 20%)and extracted with CHCl 3(3Â10mL).The combined organic phase were washed with brine,dried over Na 2SO 4,and then concentrated in vacuo.The res-idue was purified by crystallization from suitable solvent (see Sup-plementary data).5.4.General procedure for the synthesis of thioflavones 4a–p A 100mL flask was charged with flavones 3a–p (0.05mol)and Lawesson’s reagent (0.025mol),whereupon the temperature of the reaction mixture increases to 78–80°C.After 5min,35mL of ben-zene was added,the mixture was heated at refluxing for 2h,then was cooled to r.t.,and the solvent removed under vacuum.The crude solid was purified by suitable method (see Supplementary data).5.5.Determination of hMAO isoform activityThe effects of the test compounds on hMAO isoform enzymatic activity were evaluated by a fluorimetric method following the experimental protocol previously described.28Briefly,0.1mL of sodium phosphate buffer (0.05M,pH 7.4)con-taining various concentrations of the test drugs (new compounds or reference inhibitors)and adequate amounts of recombinant hMAO-A or -B required and adjusted to obtain in our experimental conditions the same reaction velocity,i.e.,to oxidize (in the control group)165pmol of p -tyramine/min (hMAO-A:1.1l g protein;spe-cific activity:150nmol of p-tyramine oxidized to p -hydroxyphe-nylacetaldehyde/min/mg protein;hMAO-B:7.5l g protein;specific activity:22nmol of p -tyramine transformed/min/mg protein)were incubated for 15min at 37°C in a flat-black-bottom 96-well microtest plate (BD Biosciences,Franklin Lakes,NJ,USA)placed in the dark multimode microplate reader chamber.After this incubation period,the reaction was started by adding(finalFigure 2.Superimposition of best fully energy minimized hMAO-B poses of (R )-2j and (S )-2j .The ligand is reported as white and green carbon polytube,respectively.The interacting residues with (R )-2j and (S )-2j are colored in white and green,respectively.The FAD cofactor is displayed as a spacefill structure.Non-polar hydrogen atoms are omitted for clarity.F.Chimenti et al./Bioorg.Med.Chem.18(2010)1273–12791277concentrations)200l M Amplex Red reagent,1U/ml horseradish peroxidase and1mM p-tyramine.The production of H2O2and, consequently,of resorufin was quantified at37°C in a multimode microplate reader(Fluostar Optima,BMG Labtech GmbH,Offen-burg,Germany),based on thefluorescence generated(excitation, 545nm,emission,590nm)over a15min period,in which thefluo-rescence increased linearly.Control experiments were carried out simultaneously by replac-ing the test drugs(new compounds and reference inhibitors)with appropriate dilutions of the vehicles.In addition,the possible capacity of the above test drugs to modify thefluorescence gener-ated in the reaction mixture due to non-enzymatic inhibition(e.g. for directly reacting with Amplex Red reagent)was determined by adding these drugs to solutions containing only the Amplex Red re-agent in a sodium phosphate buffer.The specificfluorescence emission(used to obtain thefinal re-sults)was calculated after subtraction of the background activity, which was determined from vials containing all components ex-cept the MAO isoforms,which were replaced by a sodium phos-phate buffer solution.5.6.Reversibility and irreversibility experimentsTo evaluate whether some of the tested compounds,2j,(R)-2j, and(S)-2j,are reversible or irreversible hMAO-B inhibitors,an effective centrifugation–ultrafiltration method(so-called repeated washing)previously described was used.26Briefly,adequate amounts of the recombinant hMAO-B were incubated together with a single concentration(see Table4)of the test drugs or the reference inhibitor R-(À)-deprenyl in a so-dium phosphate buffer(0.05M,pH7.4)for15min at37°C.Then,an aliquot of this incubated was stored at4°C and used for subsequent measurement of hMAO-B activity under the exper-imental conditions indicated above(see the subsection determina-tion of MAO activity).The remaining incubated sample(300l L) was placed in a Ultrafree-0.5centrifugal tube(Millipore,Billerica, USA)with a30kDa Biomax membrane in the middle of the tube and centrifuged(9000g,20min,4°C)in a centrifuge(J2-MI,Beck-man Instruments,Inc.,Palo Alto,California,USA).The enzyme re-tained in the30kDa membrane was resuspended in sodium phosphate buffer at4°C and centrifuged again(under the same experimental conditions described above)two successive times. After the third centrifugation,the enzyme retained in the mem-brane was resuspended in sodium phosphate buffer(300l L)and an aliquot of this suspension was used for subsequent hMAO-B activity determination.Similar studies were carried out on hMAO-A activity in presence of the reference inhibitor moclobemide under the experimental conditions described above.Control experiments were performed simultaneously(to define 100%hMAO activity)by replacing the test drugs with appropriate dilutions of the vehicles.The corresponding values of percent(%) hMAO inhibition were separately calculated for samples with and without repeated washing.6.Enantioseparation studiesHPLC enantioseparations were performed by using stainless-steel Chiralpak IA(250Â4.6mm I.D.and250Â10mm I.D.) (Daicel,Chemical Industries,Tokyo,Japan)columns.HPLC-grade solvents were supplied by Carlo Erba(Milan,Italy).HPLC apparatus consisted in a Perkin–Elmer(Norwalk,CT,USA)200lc pump equipped with a Rheodyne(Cotati,CA,USA)injector,a HPLC Dio-nex(CA,USA)Model TCC-100oven and a Jasco(Jasco,Ishikawa-cho,Hachioji City,Tokyo,Japan)Model2095Plus UV/CD detector.The mobile phases werefiltered and degassed by sonication imme-diately before using.In analytical enantioseparations,standard solutions were prepared by dissolving about1mg of racemic sam-ple,into10ml of methanol.The injection volume was10–20l l.In a semipreparative enantioseparation1mL sample loop was used. After semipreparative separation,the collected fractions were ana-lyzed by chiral analytical columns to determine their enantiomeric excess(ee).The column hold-up time(t0=3.0min for the250Â4.6mm i.d. column)was determined from the elution of an unretained marker (toluene),using methanol as eluent,delivered at aflow-rate of 1.0mL/min.The chromatographic and polarimetric data of the enantiomers separated at semipreparative scale are summarized in Supplemen-tary data,Table4.Specific rotations were measured at589nm by a Perkin-Elmer polarimeter model241equipped with a Na lamp.The volume of the cell was1mL and the optical path was10cm.The system was at a temperature of20°C by a Neslab RTE740cryostat. The circular dichroism(CD)spectra of the enantiomers of2f,2g, 2h,2j,2k,and2n,dissolved in ethanol(concentration about 0.2mg/mL),in a quartz cell(0.1cm-path length)at25°C,were measured by using a Jasco Model J-700spectropolarimeter(see Supplementarydata,Fig.1).The spectra are average computed over three instrumental scans and the intensities are presented in terms of ellipticity values(mdeg).7.Molecular modelingThe Protein Data Bank34(PDB)crystallographic structures 2Z5X35and2BK336were considered as receptor model of hMAO-A and-B,respectively.Both(R)and(S)enantiomers of2j were built by means of the Maestro GUI37and energy minimized using the OPLS-AA38force field as implemented in Macromodel ver.7.2.39Water solvent ef-fects were taken into account using the implicit solvation model GB/SA.40The optimized structures were submitted to docking sim-ulations with respect to hMAO-A and-B PDB crystallographic structures.Both receptor models required graphical manipulation: the co-crystallized ligands,harmine and farnesol,respectively,for 2Z5X and2BK3,were removed,FAD double bonds were corrected, and hydrogen atoms were added onto both proteins and cofactors.According to the Glide41methodology,a regular box,of about 110,000Å3,centered onto the cofactor N5atom,was considered as the enzyme active site for both hMAO-A and-B models.In order to take into account the inducedfit phenomena,2j enantiomers were docked using the Glide‘‘flexible”algorithm.The binding affinity has been evaluated using the G Score average value com-puted from the ten best generated poses for each enantiomer into the hMAO-A and–B,respectively(see Supplementary data).The most stable complexes were submitted to energy minimization using the same forcefield and aqueous environment previously re-ported.The resulting optimized structures were considered for the binding modes graphical analysis.PyMol ver.0.9842was used to create Figs.1and2.The results obtained in this study indicate thatflavonoids may have interesting therapeutic potential as original chemical models (templates)for the design and subsequent development of new drugs(selective and efficient MAO-B inhibitors)useful for improv-ing the pharmacological treatment of neurodegenerative diseases (e.g.Parkinson’s disease).AcknowledgmentsThis work was supported by Grants from MURST(Italy).Minis-terio de Sanidad y Consumo(Spain;FISS PI061537)and Consellería1278 F.Chimenti et al./Bioorg.Med.Chem.18(2010)1273–1279。