Screening and Characterization of Bioactive Molecules Derived From Medicinal Plants for Antileishmanial Activities for Leishmania donovani

Arushdeep Sidana, Umar Farooq, Amir Khan, Shiwani Kausal
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Abstract

Abstract- Leishmaniasis is a vector-borne disease caused by genus Leishmania. It causes significant morbidity and mortality in the endemic areas of several developing countries. Due to multidrug resistance in Leishmania sp and unavailability of an effective vaccine, discovery of new drugs is urgently needed. The aim of the present study was screening of medicinal plants used as Indian traditional medicine for leishmanicidal activity. Promastigote forms of Leishmania parasite were cultured in-vitro in NNN medium and further sub-cultured and maintained in RPMI-1640 medium containing 10% Fetal Bovine Serum for the screening of medicinal plants. A total of 26 medicinal plants were collected and screened for leishmanicidal activity. The methanolic extracts showing antileishmanial activity were subjected to LC-MS analysis to identify the major phyto-constituents in the crude methanolic extracts. The compounds were detected in the LC-MS of active extracts. The plant extract showing maximum antileishmanial activity was further fractionated to isolate the major compound(s).The compound isolated was characterised by IR, Mass spectrometry and subjected to in vitro antileishmanial activity against L. donovani promastigotes. In vitro antileishmanial assay revealed that crude methanolic extracts of 2/10 plants were active against L. donovani promastigotes. Methanolic root extracts of Inula racemosa were found active (54.83%) against the parasite while T. Terrestris were found least active with such as toxicity, percent inhibition 43.10% at concentration of 500 μg/ml. Inula racemosa methanolic extract further fractionated subjected for LC-MS analysis. The compound isolated from the methanolic root extract of I. racemosa was isoalantolactone, which did not show any antileishmanial activity against L. donovani. The study suggested that crude extract of Inula racemosa and T. Terrestris have shown potent antileishmanial activity while extracted bioactive molecules does not show efficacy against Leishmania parasite. Hence the antileishmanial activity could be due to any other compound which could not be detected, so further study is undertaken.
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药用植物抗多诺瓦利什曼原虫活性分子的筛选与鉴定
摘要-利什曼病是由利什曼属引起的媒介传播疾病。它在一些发展中国家的流行地区造成了很高的发病率和死亡率。由于利什曼原虫的多药耐药和缺乏有效的疫苗,迫切需要发现新的药物。本研究的目的是筛选作为印度传统药物的药用植物的利什曼尼杀灭活性。在NNN培养基中体外培养利什曼原虫的Promastigote形式,并在含有10%胎牛血清的rmi -1640培养基中进一步继代培养和维持,以筛选药用植物。共收集26株药用植物进行利什曼尼杀虫活性筛选。对具有抗利什曼原虫活性的甲醇提取物进行LC-MS分析,鉴定其主要植物成分。活性提取物的LC-MS检测到化合物。对具有最大抗利什曼活性的植物提取物进行进一步分离,以分离出主要化合物。对分离得到的化合物进行了红外光谱、质谱鉴定,并对其体外抗多诺瓦氏L. promastigotes的利什曼原虫活性进行了测试。体外抗利什曼原虫实验表明,2/10的植物粗甲醇提取物对多诺瓦螺旋体具有抗利什曼原虫活性。总状菊甲醇根提取物在500 μg/ml浓度下对寄生虫的抑制率为43.10%,活性最低(54.83%);总状菊甲醇提取物进一步分离,进行LC-MS分析。从总状花甲醇根提取物中分离得到的化合物为异丙酸内酯,该化合物对多氏利什曼菌无抗利什曼活性。研究表明,总状花序和地鸡翅的粗提物具有较强的抗利什曼原虫活性,而提取的生物活性分子对利什曼原虫没有活性。因此,抗利什曼原虫活性可能是由于任何其他化合物无法检测到,因此进行了进一步的研究。
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