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In vitro, in silico and in vivo studies of ursolic acid as an anti-filarial agent.

Kalani K, Kushwaha V, Sharma P, Verma R, Srivastava M, Khan F, Murthy PK, Srivastava SK - PLoS ONE (2014)

Bottom Line: The selectivity index (SI) of UA for the parasites was found safe.Thus, in conclusion in vitro, in silico and in vivo results indicate that UA is a promising, inexpensive, widely available natural lead, which can be designed and developed into a macrofilaricidal drug.To the best of our knowledge this is the first ever report on the anti-filarial potential of UA from E. tereticornis, which is in full agreement with the Thomson Reuter's 'Metadrug' tool screening predictions.

View Article: PubMed Central - PubMed

Affiliation: Medicinal Chemistry Department, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow, 226015 (U.P.) India; Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, New Delhi, 110 001, India.

ABSTRACT
As part of our drug discovery program for anti-filarial agents from Indian medicinal plants, leaves of Eucalyptus tereticornis were chemically investigated, which resulted in the isolation and characterization of an anti-filarial agent, ursolic acid (UA) as a major constituent. Antifilarial activity of UA against the human lymphatic filarial parasite Brugia malayi using in vitro and in vivo assays, and in silico docking search on glutathione-s-transferase (GST) parasitic enzyme were carried out. The UA was lethal to microfilariae (mf; LC100: 50; IC50: 8.84 µM) and female adult worms (LC100: 100; IC50: 35.36 µM) as observed by motility assay; it exerted 86% inhibition in MTT reduction potential of the adult parasites. The selectivity index (SI) of UA for the parasites was found safe. This was supported by the molecular docking studies, which showed adequate docking (LibDock) scores for UA (-8.6) with respect to the standard antifilarial drugs, ivermectin (IVM -8.4) and diethylcarbamazine (DEC-C -4.6) on glutathione-s-transferase enzyme. Further, in silico pharmacokinetic and drug-likeness studies showed that UA possesses drug-like properties. Furthermore, UA was evaluated in vivo in B. malayi-M. coucha model (natural infection), which showed 54% macrofilaricidal activity, 56% female worm sterility and almost unchanged microfilaraemia maintained throughout observation period with no adverse effect on the host. Thus, in conclusion in vitro, in silico and in vivo results indicate that UA is a promising, inexpensive, widely available natural lead, which can be designed and developed into a macrofilaricidal drug. To the best of our knowledge this is the first ever report on the anti-filarial potential of UA from E. tereticornis, which is in full agreement with the Thomson Reuter's 'Metadrug' tool screening predictions.

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Micro-(A) and macrofilaricidal (B) activity of UA and reference drug diethylcarbamazine-citrate (DEC-C) against Brugia malayi in Mastomys coucha.Values are mean ± S.D. of 5 animals from two experiments. (A) No alteration in Microfilarial count in treated animals at each time point post initiation of treatment over day 0. Statistics: Student's ‘t’ test. Significance level (B) *P<0.05 (vs sterilized female worm of control animals).
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pone-0111244-g005: Micro-(A) and macrofilaricidal (B) activity of UA and reference drug diethylcarbamazine-citrate (DEC-C) against Brugia malayi in Mastomys coucha.Values are mean ± S.D. of 5 animals from two experiments. (A) No alteration in Microfilarial count in treated animals at each time point post initiation of treatment over day 0. Statistics: Student's ‘t’ test. Significance level (B) *P<0.05 (vs sterilized female worm of control animals).

Mentions: Microfilaricidal activity: Figure 5A shows anti-filarial efficacy of UA against B. malayi in M. coucha at 100 mg/kg s.c. for 5 consecutive days. UA produced 4–33% lower microfilaremia (statistically not significant) than 0 day throughout the post treatment observation period (Figure 5A). In other words, the microfilaremia in UA treated animals remained below (4–33%) the pretreatment (0 day) level throughout the observation period, while in the untreated control it was (progressively) higher than the pretreatment level and never equaled the 0 day level. This clearly shows that UA possesses considerable antifilarial efficacy. DEC-C (50 mg/kg, s.c. x 5 days), which is principally a microfilaricide, caused >85% reduction in microfilarial count on day 7 p.i.t. which progressively increased and relapsed on day 49 p.i.t.; the count further increased rapidly and crossed the pretreatment level by day 56 p.i.t. The trend of microfilaremia from day 7 to day 84 p.i.t. in the three groups (control untreated, UA treated and DEC treated) against time (post treatment) was determined and compared among each other using linear trend analysis. The baseline of each group was subjected to equality and the observations were converted to show percent change at each time point. The baseline adjusted data was fitted to straight line. The analysis showed that while the mf count in the DEC-treated animals after an initial dramatic drop on day 7 p.i.t., increased gradually over time, it remained almost unchanged with time in UA treated group (trend not significant). Thus UA was found to be better than DEC in controlling microfilaremia. The details are given in File S1: ‘In vivo antifilarial efficacy in Brugia malayi -M. coucha model: Microfilaricidal activity’.


In vitro, in silico and in vivo studies of ursolic acid as an anti-filarial agent.

Kalani K, Kushwaha V, Sharma P, Verma R, Srivastava M, Khan F, Murthy PK, Srivastava SK - PLoS ONE (2014)

Micro-(A) and macrofilaricidal (B) activity of UA and reference drug diethylcarbamazine-citrate (DEC-C) against Brugia malayi in Mastomys coucha.Values are mean ± S.D. of 5 animals from two experiments. (A) No alteration in Microfilarial count in treated animals at each time point post initiation of treatment over day 0. Statistics: Student's ‘t’ test. Significance level (B) *P<0.05 (vs sterilized female worm of control animals).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4222910&req=5

pone-0111244-g005: Micro-(A) and macrofilaricidal (B) activity of UA and reference drug diethylcarbamazine-citrate (DEC-C) against Brugia malayi in Mastomys coucha.Values are mean ± S.D. of 5 animals from two experiments. (A) No alteration in Microfilarial count in treated animals at each time point post initiation of treatment over day 0. Statistics: Student's ‘t’ test. Significance level (B) *P<0.05 (vs sterilized female worm of control animals).
Mentions: Microfilaricidal activity: Figure 5A shows anti-filarial efficacy of UA against B. malayi in M. coucha at 100 mg/kg s.c. for 5 consecutive days. UA produced 4–33% lower microfilaremia (statistically not significant) than 0 day throughout the post treatment observation period (Figure 5A). In other words, the microfilaremia in UA treated animals remained below (4–33%) the pretreatment (0 day) level throughout the observation period, while in the untreated control it was (progressively) higher than the pretreatment level and never equaled the 0 day level. This clearly shows that UA possesses considerable antifilarial efficacy. DEC-C (50 mg/kg, s.c. x 5 days), which is principally a microfilaricide, caused >85% reduction in microfilarial count on day 7 p.i.t. which progressively increased and relapsed on day 49 p.i.t.; the count further increased rapidly and crossed the pretreatment level by day 56 p.i.t. The trend of microfilaremia from day 7 to day 84 p.i.t. in the three groups (control untreated, UA treated and DEC treated) against time (post treatment) was determined and compared among each other using linear trend analysis. The baseline of each group was subjected to equality and the observations were converted to show percent change at each time point. The baseline adjusted data was fitted to straight line. The analysis showed that while the mf count in the DEC-treated animals after an initial dramatic drop on day 7 p.i.t., increased gradually over time, it remained almost unchanged with time in UA treated group (trend not significant). Thus UA was found to be better than DEC in controlling microfilaremia. The details are given in File S1: ‘In vivo antifilarial efficacy in Brugia malayi -M. coucha model: Microfilaricidal activity’.

Bottom Line: The selectivity index (SI) of UA for the parasites was found safe.Thus, in conclusion in vitro, in silico and in vivo results indicate that UA is a promising, inexpensive, widely available natural lead, which can be designed and developed into a macrofilaricidal drug.To the best of our knowledge this is the first ever report on the anti-filarial potential of UA from E. tereticornis, which is in full agreement with the Thomson Reuter's 'Metadrug' tool screening predictions.

View Article: PubMed Central - PubMed

Affiliation: Medicinal Chemistry Department, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow, 226015 (U.P.) India; Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, New Delhi, 110 001, India.

ABSTRACT
As part of our drug discovery program for anti-filarial agents from Indian medicinal plants, leaves of Eucalyptus tereticornis were chemically investigated, which resulted in the isolation and characterization of an anti-filarial agent, ursolic acid (UA) as a major constituent. Antifilarial activity of UA against the human lymphatic filarial parasite Brugia malayi using in vitro and in vivo assays, and in silico docking search on glutathione-s-transferase (GST) parasitic enzyme were carried out. The UA was lethal to microfilariae (mf; LC100: 50; IC50: 8.84 µM) and female adult worms (LC100: 100; IC50: 35.36 µM) as observed by motility assay; it exerted 86% inhibition in MTT reduction potential of the adult parasites. The selectivity index (SI) of UA for the parasites was found safe. This was supported by the molecular docking studies, which showed adequate docking (LibDock) scores for UA (-8.6) with respect to the standard antifilarial drugs, ivermectin (IVM -8.4) and diethylcarbamazine (DEC-C -4.6) on glutathione-s-transferase enzyme. Further, in silico pharmacokinetic and drug-likeness studies showed that UA possesses drug-like properties. Furthermore, UA was evaluated in vivo in B. malayi-M. coucha model (natural infection), which showed 54% macrofilaricidal activity, 56% female worm sterility and almost unchanged microfilaraemia maintained throughout observation period with no adverse effect on the host. Thus, in conclusion in vitro, in silico and in vivo results indicate that UA is a promising, inexpensive, widely available natural lead, which can be designed and developed into a macrofilaricidal drug. To the best of our knowledge this is the first ever report on the anti-filarial potential of UA from E. tereticornis, which is in full agreement with the Thomson Reuter's 'Metadrug' tool screening predictions.

Show MeSH
Related in: MedlinePlus