A new technique of in vitro assay of antifilarials using different life-forms of Acanthocheilonema viteae.

D P Singh, S Misra, R K Chatterjee
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Abstract

An approach has been made to develop an in vitro screening system to evaluate antifilarial efficacy of compounds and an effort has been made to establish correlation between in vivo and in vitro screening technique. The in vitro experiments were conducted simultaneously using three life-forms (adult, microfilaria and infective larva) of Acanthocheilonema viteae using five antifilarial agents representing four chemical groups. All the selected antifilarials were known to be active against one or more life-stages of human lymphatic or animal filariids. Diethylcarbamazine and Centperazine showed 100% microfilaricidal and infective larvicidal actions at concentrations of 0.5 and 0.25 mg/ml and 0.5 and 0.0313 mg/ml respectively with no effect on adult worms even at 1 mg/ml. Levamisole was effective against all the three life-stages killing 100% adult worms at 1 mg/ml, infective larvae at 0.0625 mg/ml and microfilariae at 0.0125 mg/ml, while mebendazole exhibited activity only against adult worms (100% at 0.5 mg/ml). Ivermectin killed adult females and microfilariae at 0.063 and 0.5 mg/ml respectively but did not affect infective larvae even up to 1 mg/ml concentration. The study indicated that in vitro screening system can be used for primary screening of potential antifilarial agents provided three life-forms of A. viteae are used simultaneously to avoid false negative results. It would however be more appropriate if a few compounds of a particular chemical class are initially assessed both in vivo and in vitro for validity of subsequent test results in vitro.

利用不同生命形式的棘条线虫体外测定抗丝虫药的新技术。
建立了一种体外筛选系统来评价化合物的抗丝效果,并努力建立体内筛选技术和体外筛选技术之间的相关性。采用4个化学类群的5种抗丝剂,同时对三种生活型(成虫、微丝虫和感染性幼虫)进行体外实验。已知所有选定的抗丝虫药对人淋巴丝虫病或动物丝虫病的一个或多个生命阶段都有活性。二乙基卡马嗪和噻嗪在0.5和0.25 mg/ml浓度和0.5和0.0313 mg/ml浓度下分别表现出100%的杀微丝和杀幼虫效果,即使在1 mg/ml浓度下对成虫也没有效果。左旋咪唑在1 mg/ml、0.0625 mg/ml和0.0125 mg/ml浓度下均能杀灭100%的成虫,而甲苯咪唑仅在0.5 mg/ml浓度下杀灭100%的成虫。伊维菌素浓度为0.063 mg/ml和0.5 mg/ml时,对雌虫和微丝虫均有杀伤作用,但当浓度达到1 mg/ml时,对感染幼虫无杀伤作用。研究表明,如果同时使用三种生命形式的黄曲霉进行体外筛选,可用于潜在抗丝虫药的初步筛选,避免假阴性结果。然而,如果首先在体内和体外对某一特定化学类别的几种化合物进行评估,以确定随后的体外测试结果的有效性,则更为合适。
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