Study on Flavonoids from the Methanol Extracts of Tephrosia purpurea Leaves

[Objective] The aim was to determine flavonoids from the MeOH extracts of Tephrosia purpurea leaves and their cytotoxicitives against the ovarian cells from Sprodenia litura （SL cells）.[Method] The compounds were isolated with column chromatography and their structures were established on the basis of various spectroscopic analysis （including UV,1D and 2D NMR analyses as well as HR-ESRMS）.The cytotoxicity against the SL cells was evaluated by using MTT method.[Result] Six known flavonoids,6-methoxykaempferol （1）,6-methoxykaempferol 7-O-α-rhamnopyranoside （2）,6-methoxykaempferol 3-O-α-rhamnopyranosyl（1→2）[α-rhamno-pyranosyl（1→6）]-β-galactopyranoside （3）,6-methoxykaempferol 3-O-α-rhamnopyranosyl （1 →2）[α-rhamnopyranosyl （1 →6）]-β-galactopyranoside-7-O-α-rhamnopyranoside （4）,pongachin （5）,5,7-dimethoxy-8-（3-hydroxy-3-methylbut-1Z-enyl） flavanone （6） were isolated and determined.Except compound 5,the others were isolated from T.purpurea for the first time.For the cytotoxicity compound 5 had significant activity with the IC50 value of 4.4 mg/L while compound 1 and 3,whose cytotoxicity exceeded rotenone,also showed moderate activity.[Conclusion] Of all the compounds from T.purpurea leaves,the content of 6-methoxykaempferol compounds was considerable.The profiles of these compounds against SL cells suggested that compounds 1,3 and 5,whose cytotoxicity exceeded rotenone,were worth further research.

灰毛豆Tephrosia purpurea中查尔酮类化合物的分离鉴定

【目的】为探讨灰毛豆属植物Tephrosia purpurea化学成分中的查尔酮类化合物,对其枝、叶部位进行化学成分分离及结构解析。【方法】采用乙醇提取、溶剂萃取、柱层析分离方法得到多种灰毛豆属T.purpurea化学成分,并用波谱分析手段对其进行结构表征。【结果】从灰毛豆属T.purpurea中共分离鉴定出11个查尔酮类化合物,分别为elatadihydrochalcone(1),spinochalcone A(2),3′,5′-diisopentenyl-2′,4′-dihydroxychalcone(3),candidachalcone(4),O-methylpongamol(5),(+)-tephrosone(6),spinochalcone C(7),epoxyobovatachalcone(8),tunicatachalcone(9),crassichalone(10),(S)-elatadihydrochalcone(11)。【结论】以上化合物均为查尔酮,且均属首次从该植物中分离得到。

Anticancer activity of Tephrosia purpurea and Ficus religiosa using MCF 7 cell lines

Objective:To investigate anticancer activity of different fractions of Tephrosia purpurea[TP] （Sharapunkha,Fabaceae） and Ficus religiosa[FR]（Peepal,Moraceae）.Methods:The fractions of TP and FR were prepared and tested for in vitro anticancer activity using human MCF 7 cell line by trypan blue exclusion method.Results:The result showed that among all these fractions of TPI.TPIII.FRI and FRIII showed better anticancer activity compared to other fractions.The IC50 value for TPI（152.4μM）,TPIII（158.71μM）.FRI（160.3μM） and for FRIII（222.7μM） was observed.Conclusions:The present study shows anticancer potential of TP and FR fractions in MCF 7 cell line.

Evaluation of antiinflammatory activity of Tephrosia purpurea in rats

Objective:To evaluate the antiinflammatory activity of orally administered ethanolic extract of Tephrosia purpurea in acute and subacute inflammation in rats.Methods:An ethanolic extract of Tephrosia purpurea was prepared.Carrageenan induced paw edema and cotton pellet granuloma were the models for acute and subacute inflammation respectively.Four groups of rats in each model were treated orally with 2%gum acacia,100 mg /kg of aspirin,500 mg/kg and 1 000 mg/kg of ethanolic extract of Tephrosia purpurea respectively.In carrageenan induced paw edema model, subplantar injection of 1%carrageenan was made into the hind paw of the rats sixty minutes after the administration of the respective drugs.The paw volume was measured immediately after injection of carrageenan,at 3 hours and at 6 hours.Then percentage inhibition of edema was calculated.In the cotton pellet granuloma model,animals were administered drugs for six days after placing cotton pellets in the axilla on each side.On the 7th day,dry weight of granuloma was calculated.Results:The rats treated with Tephrosia purpurea did not exhibit any significant decrease in paw volume and serum ceruloplasmin levels as compared to the control and aspirin treated groups in the acute inflammation model;while,there was a significant(P 【 0.01) decrease in the weight of granuloma in Tephrosia purpurea and aspirin treated groups as compared to control in subacute inflammation.Conclusions:The ethanolic extract of orally administered Tephrosia purpurea shows significant antiinflammatory effect in subacute inflammation but not in acute inflammation in rats.

Ethnopharmacology, phytochemistry and pharmacology of Tephrosia purpurea

Tephrosia purpurea(L.) Pers. is popularly known as ‘Sarapunkha' in classical Ayurvedic texts. It is a perennial plant belonging to the family Fabaceae, and occurs throughout the Indian subcontinent. T. purpurea is traditionally used to treat spleenomegaly, cirrhosis, cough and cold, abdominal swelling and as an antidote in the Ayurvedic system of medicine. Phytochemical investigations indicate the presence of semiglabrin, pongamole, lanceolatins A and B, rutin, lupeol, and β-sitosterol. Flavonoids including(+)-tephrorin A and B,(+)-tephrosone, an isoflavone, 7, 4'-dihydroxy-3', 5'-dimethoxyisoflavone and a chalcone,(+)-tephropurpurin were isolated from the whole plant. Pharmacological activities of different parts of the plant reported include anti-inflammatory, antiulcer, antimicrobial, antioxidant, antiallergic, antidiabetic, hepatoprotective, antitumor and insect repellent activity. In the present review, the literature on the phytochemical and pharmacological investigations of Tephrosia purpurea(L.) Pers. are summarized to August, 2012.

Therapeutic potential of ethyl acetate fraction of Tephrosia purpurea Linn.leaves in a rat model of gout

Objective: The present study is to determine the potential treatment effects of ethyl acetate fraction of Tephrosia purpurea Linn. leaves(EATP) against gout.Methods: Gout in experimental rats was induced with potassium oxonate at the dose of 250 mg/kg(intraperitoneal injection) for 7 consecutive days;EATP was administered 1 h after administration of the potassium oxonate on each day of experiment. Potassium oxonate was discontinued on the 8 th day;thereafter allopurinol(10 mg/kg, p.o.) and EATP(200 and 400 mg/kg, p.o.) were continued until day 14. The uric acid level was measured from serum and urine during the experiment. Other biochemical parameters were assessed, including blood and urine creatinine, erythrocyte sedimentation rate, and total protein. Blood urea nitrogen, serum aspartate aminotransferase serum alanine aminotransferase and alkaline phosphatase were also measured. The blood was analyzed for levels of malondialdehyde and the antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase.Histopathological and radiological changes in the ankle of rats were observed after completion of the experiment.Results: EATP was able to decrease serum uric acid and creatinine level;it also reduced inflammation,oxidative stress and lysosomal enzyme level, which has a role in acute inflammation. EATP increased uric acid excretion through urine due to its uricosuric effect.Conclusion: EATP lowered the serum uric acid level and increased the urine uric acid level through excretion, which is useful in the treatment of gout. Hence the EATP was found to be helpful in the treatment of gout.

灰毛豆精油对谷蠹的毒杀作用

为评价灰毛豆叶、树皮和种子精油对谷蠹的毒杀活性,分别测定了它们对谷蠹的驱避活性、触杀活性、熏蒸活性及种群抑制作用。结果表明,处理48 h后,3种精油对谷蠹成虫的驱避率达到最大值,分别是82.3%、90.7%和94.3%;3种精油都对谷蠹5日龄若虫具有触杀活性,其LD50值分别为每虫25.0、27.9和27.8 mg;在用量为20.0 mg/L时,3种精油对谷蠹成虫都具有熏蒸活性,熏蒸48 h后成虫的死亡率为56.2%~57.4%,3种精油的熏蒸活性无显著性差异;当100 g稻谷中拌有200 mg精油时,3种精油对当代种群的抑制率分别为45.5%、53.6%和76.8%,对子代种群的抑制率分别为43.8%、68.7%和75.1%,其中种子精油的抑制活性最高,且显著高于树皮及叶片精油的活性。

灰毛豆种子甲醇提取物对黑肩绿盲蝽成虫的安全性

为评价灰毛豆种子甲醇提取物对白背飞虱天敌黑肩绿盲蝽的安全性,比较了提取物对黑肩绿盲蝽成虫和白背飞虱3龄若虫的杀虫毒力,测定了提取物对黑肩绿盲蝽成虫存活时间和捕食功能的影响。结果表明,提取物对黑肩绿盲蝽成虫的毒力远低于对白背飞虱3龄若虫的毒力,处理后48 h的LC50值分别为298.8和16.3 mg/L;用10 mg/L的提取物处理后,黑肩绿盲蝽成虫对白背飞虱3龄若虫密度变化的功能反应符合Holling-II功能反应模型,处理后第1天的捕食能力增强,第2、3天捕食能力逐日下降;黑肩绿盲蝽成虫接触提取物后,存活时间为13.3 d,显著低于未接触提取物的存活时间21.3 d。综合试验结果,灰毛豆种子甲醇提取物对黑肩绿盲蝽成虫不安全。

灰毛豆叶片的化学成分研究

从灰毛豆叶片中分离得到10个化合物,通过波谱数据和理化性质分别鉴定为:stigmast-1,5-dien-3β-ol(1),clemaphenol A(2),松脂醇(3),桉素(4),3-吲哚甲醛(5),色氨酸(6),(S)-4-苄基-2-噁唑烷酮(7),4-羟基苯基丙酸(8),buteaspermanol(9),bovatachalcone(10)。其中化合物1、2、4、5、7、8、9首次从该植物中分离得到。

灰毛豆树皮中的杀虫成分及其杀虫活性

为确定灰毛豆Tephrosia purpurea树皮甲醇提取物中的杀虫成分,以白纹伊蚊Aedes albopictus 4龄幼虫为靶标昆虫,在活性跟踪的基础上利用色谱技术分离其活性成分,然后根据各化合物的核磁共振图谱和质谱数据确定化合物的结构,并利用玻片载蚜法和点滴法测定了各化合物对桃蚜Myzus persicae无翅蚜成虫和小菜蛾Pluttella xylostella 3龄幼虫的毒杀活性。结果表明：从该植物树皮甲醇提取物中共分离、鉴定了10个对白纹伊蚊幼虫具有毒杀作用的化合物,即12a-羟基鱼藤酮（12a-hydroxyrotenone）,4’-hydroxyemoroidocarpan,豆薯内酯（pachyrrhizine）,鱼藤酮（rotenone）,6-甲氧基香豆素（6-methoxycoumarin）,（-）-edunol,obovatin,pongachin,12-acetyelliptinol和2-hydroxyrotenone。这些化合物对该蚊虫幼虫处理24 h的LC50值分别是12.5,22.1,25.0,34.1,43.4,58.4,121.9,191.0,219.8和250.0 mg/L。3个化合物（4’-hydroxyemoroidocarpan,鱼藤酮和12a-羟基鱼藤酮）对桃蚜成虫和小菜蛾3龄幼虫表现出毒杀活性,它们对桃蚜24 h的LC50值分别是49.9,1.9和0.9 mg/L,对小菜蛾幼虫24 h的LD50值分别是49.8,197.1和40.9μg/头。首次从该植物中分离得到6个已知的黄酮类化合物[4’-hydroxyemoroidocarpan,2-hydroxyrotenone,（-）-edunol,12-acetyelliptinol,pongachin和obovatin]和2个已知的香豆素类化合物（6-甲氧基香豆素和豆薯内酯）。阐明这些杀虫化合物的结构不仅有利于理解植物和昆虫的关系,而且有助于评价该植物及其活性化合物作为植物源农药开发利用的潜力。

灰毛豆甲醇提取物的杀虫活性

研究灰毛豆（Tephrosia purpurea）各部位提取物的杀虫活性及其作用方式。结果表明,灰毛豆对白纹伊蚊Aedes albopictus Skuse幼虫、菜青虫Pieris rapae（L.）、斜纹夜蛾Prodenialitura（Fabricius）幼虫和黄曲条跳甲Phyllotreta striolata（Fabricius）成虫都有杀虫活性。灰毛豆种子、树皮、根皮、豆荚、枝条和树叶的甲醇提取物对白纹伊蚊4龄幼虫24h的LC50值分别是22.1,97.7,36.6,142.6,165.6和618.3mg.L-1,树干木质部没有杀虫活性;灰毛豆种子、树皮和根皮甲醇提取物对3龄菜青虫24h的触杀毒力LC50值分别是232.1,206.3和236.7mg.L-1;种子、树皮、根皮、枝条、树叶和豆荚对3龄菜青虫的24h胃毒毒力LC50值分别是192.6,168.4,249.7,524.5,1001.0,和510.7mg.L-1。在取食或接触有提取物的叶碟后,5龄菜青虫会出现取食量减少和生长发育变慢的亚致死现象。这些研究结果表明,灰毛豆除茎干木质部以外的其它各部位均含有杀虫活性成分,其作用方式为胃毒和触杀。

灰毛豆的化学成分研究

目的探讨灰毛豆属植物灰毛豆Tephrosia purpurea枝叶部位的化学成分。方法采用常用硅胶柱和凝胶色谱色谱手段和核磁共振谱与质谱分析方法以及通过对照文献,对灰毛豆属植物T.purpurea的枝叶部分进行了化学成分提取分离及结构鉴定。结果从灰毛豆属T.purpurea枝叶中共分离鉴定出12个化合物,分别为:pumilanol(1),tephrocandidin A(2),tephrospiroketone I(3),1β-hydroxy-6,7α-dihydroxyeudesm-4(15)-ene(4),calophione A(5),(+)-lariciresinol-9'-stearate(6),tephroapollin D(7),cineroside A(8),hookerianin(9),(2S)-5-hydroxy-7,4'-di-O-(γ,γ-dimethylallyl)flavanone(10),tephrospirolactone(11),candirone(12)。结论以上化合物均为首次从该植物中分离得到。

灰毛豆内生真菌淡紫拟青霉菌(TPL04)代谢产物的研究

从灰毛豆叶片中分离出内生真菌-淡紫拟青霉菌Purureocillium lilacinum(TPL04),对其发酵产物采用硅胶柱色谱、Sephadex LH-20凝胶色谱、制备薄层色谱、中压柱色谱及制备液相色谱等方法进行分离纯化,共获得8个单体化合物。通过波谱学方法并结合相关文献数据比对,化合物结构鉴定为5-羟基己酸-4-内酯(1)、(R)-4-苄基-2-噁唑烷酮(2)、2′-脱氧胸苷(3)、对羟基苯甲酸甲酯(4)、光色素(5)、β-胡萝卜苷(6)、尿嘧啶核苷(7)、己六醇(8)。其中化合物1、2、4是首次从淡紫拟青霉菌中分离得到,抑菌实验表明化合物4对水稻纹枯菌(Pellicularia sasakii)、辣椒疫霉菌(Phytophthora capsici)、草莓灰霉菌(Botrytis cinerea)、油菜菌核菌(Sclerotinia sclerotiorum)四种植物病原菌生长有抑制作用。

不同因素对灰毛豆树皮中12α-羟基鱼藤酮的含量影响研究

为了解灰毛豆不同品种树皮中12α-羟基鱼藤酮的含量,测定了16个品种树皮中该化合物的含量,研究了不同树龄灰毛豆树皮中12α-羟基鱼藤酮的含量变化,以及磷酸氢钙、尿素、硫酸钾对灰毛豆树皮中该化合物含量及其产量的影响。结果表明:16个品种灰毛豆树皮中12α-羟基鱼藤酮含量在0.22%~5.95%之间,其中广西2号的含量显著高于其他品种。1月播种的16个品种,12α-羟基鱼藤酮含量在11月份时接近最大值。施用磷酸氢钙(20 kg/667m2)、尿素(5 kg/667m2)和硫酸钾(5 kg/667m2)后,广西2号树皮中12α-羟基鱼藤酮的产量依次是75.33、66.34和64.26 kg/667m2,以磷酸氢钙作基肥时12α-羟基鱼藤酮的产量显著高于其他处理。广西2号品种是一种优秀的杀虫资源植物,最佳采收时期为11月份,施用磷酸氢钙有利于提高该植物中12α-羟基鱼藤酮的产量。

灰毛豆中异黄酮类、鱼藤酮类及coumestan类化合物分析

目的:探讨灰毛豆Tephrosia purpurea枝叶部位的化学成分;丰富灰毛豆属植物的化学成分数据库。方法:采用多种色谱分离手段和现代波谱分析方法以及通过对照文献,深入系统地对灰毛豆T.purpurea的枝叶部分进行了化学成分分离提取及结构鉴定。结果:从灰毛豆T.purpurea中共分离鉴定出10个异黄酮类化合物,分别为12a-dehydro-6-hydroxysumatrol(1),elongatin(2),tephcalostan C(3),7,4'-dihydroxy-3',5'-dimethoxyisoflavone(4),4'-demethyltoxicarol isoflavone(5),tephcalostan B(6),5,7-di-O-prenylbiochanin A(7),tephcalostan D(8),tephcalostan(9),6a,12a-dehydro-2,3,6-trimethoxy-8-(3',3'-dimethylallyl)-9,11-dihydroxyrotenone(10),3,4,8,9-dimethylenedioxypterocarpan(11),12a-hydroxy-β-toxicarol(12)。结论:除化合物4外,其他11个化合物均为首次从该植物中分离得到。