Bioassay-guided isolation and characterization of active antiplasmodial compounds from Murraya koenigii extracts against Plasmodium falciparum and Plasmodium berghei

Abstract

Malaria is an overwhelming impact in the poorest countries in the world due to their prevalence, virulence and drug resistance ability. Currently, there is inadequate armoury of drugs for the treatment of malaria. This underscores the continuing need for the discovery and development of new effective and safe antimalarial drugs. To evaluate the in vitro and in vivo antimalarial activity of the leaf ethyl acetate extract of Murraya koenigii, bioassay-guided chromatographic fractionation was employed for the isolation and purification of antimalarial compounds. The in vitro antimalarial activity was assayed by the erythrocytic stages of chloroquine-sensitive strain of Plasmodium falciparum (3D7) in culture using the fluorescence-based SYBR Green I assay. The in vivo assay was done by administering mice infected with Plasmodium berghei (NK65) four consecutive daily doses of the extracts through oral route following Peter's 4-day curative standard test. The percentage suppression of parasitaemia was calculated for each dose level by comparing the parasitaemia in untreated control with those of treated mice. Cytotoxicity was determined against HeLa cells using MTT assay. Histopathology was studied in kidney, liver and spleen of isolated compound-treated Swiss albino mice. The leaf crude ethyl acetate extract of M. koenigii showed good in vitro antiplasmodial activity against P. falciparum. The in vivo test of the leaf crude ethyl acetate extract (600 mg/kg) showed reduced malaria parasitaemia by 86.6% against P. berghei in mice. Bioassay-guided fractionation of the leaf ethyl acetate extract of M. koenigii led to the isolation of two purified fractions C3B2 (2.84 g) and C3B4 (1.97 g). The purified fractions C3B2 and C3B4 were found to be active with IC50 values of 10.5 ± 0.8 and 8.25 ± 0.2 μg/mL against P. falciparum, and in vivo activity significantly reduced parasitaemia by 82.6 and 88.2% at 100 mg/kg/body weight on day 4 against P. berghei, respectively. The isolated fractions C3B2 and C3B4 were monitored by thin-layer chromatography until a single spot was obtained with R f values of 0.36 and 0.52, respectively. The pure compounds obtained in the present investigation were subjected to UV-visible spectroscopy, Fourier transformer infrared spectroscopy, 1D and 2D (1)H-Nuclear magnetic resonance (NMR), (13)C NMR, DEPT, COSY and Mass spectral analysis. Based on the spectral analysis, it is concluded that the isolated compounds were myristic acid (C3B2) and β-caryophyllene (C3B4). The cytotoxic effect of myristic acid and β-caryophyllene showed the TC50 values of >100 and 80.5 μg/mL, respectively against HeLa cell line. The histopathology study showed that protection against nephrotoxicity of kidney, hepatic damage of liver and splenocytes protection in spleen was achieved with the highest dose tested at 100 mg/kg/body weight. The present study provides evidence of antiplasmodial compounds from M. koenigii and is reported for the first time.

Characterization and biotoxicity of Hypnea musciformis-synthesized silver nanoparticles as potential eco-friendly control tool against Aedes aegypti and Plutella xylostella

“…The use of synthetic insecticides to control mosquito vectors lead to high operational costs, resistance and non-target effects. In this scenario, eco-friendly control tools, such as botanical mosquitocidals, are a priority (Perry et al, 1998;Kamaraj et al, 2014;Benelli et al, 2015).…”

In Vivo Antiplasmodial Potentials of the Combinations of Four Nigerian Antimalarial Plants

“…The protoberberine alkaloids berberine, palmatine and jatrorrhizine were identified as potential active principles because they demonstrated in vitro activity against P. falciparum that was comparable ( p > 0.05) to quinine, although they lacked any in vivo activity against P. berghei [15]. Numerous monomeric and dimeric carbazole alkaloids have been reported for M. koenigii leaf [9], which has been reported to give a dose-dependent antimalarial activity against P. berghei chloroquine-resistant NK65 strain [16]. Bioassay-guided chromatographic fractionation of its leaf ethyl acetate extract, using the in vitro and chemosuppresive in vivo antimalarial assay methods, yielded myristic acid and β-caryophyllene as its antimalarial constituents [16].…”

“…Numerous monomeric and dimeric carbazole alkaloids have been reported for M. koenigii leaf [9], which has been reported to give a dose-dependent antimalarial activity against P. berghei chloroquine-resistant NK65 strain [16]. Bioassay-guided chromatographic fractionation of its leaf ethyl acetate extract, using the in vitro and chemosuppresive in vivo antimalarial assay methods, yielded myristic acid and β-caryophyllene as its antimalarial constituents [16].…”

In Vitro Assessment of Antiplasmodial and Antitrypanosomal Activities of Chloroform, Ethyl Acetate and Ethanol Leaf Extracts of Oedera genistifolia

“…In the same vein, hexadecanoic acid is known to exhibit strong antimicrobial and anti-inflammatory activity [33]. Similarly, ferruginol and caryophyllene were also present in all the extracts and their antimalarial activity has been well documented previously [1]. Therefore, fractionation and isolation of pure compounds from the crude extracts are necessary because of numerous bioactive compounds that support the medicinal application of O. genistifolia.…”

“…Malaria has been known as one of the tropical diseases with a devastating effect on the well-being of people, especially from developing countries in the world. Although considerable progress has been made in the fight to combat malaria over the last two decades, the disease burden has been prevalent in Africa, where global cases and deaths have increased to about 82% and 90%, respectively [1]. The report further signposts that children below age 5 years are more vulnerable to this disease, which account for about 78% of 90% malaria death cases reported, thus making malaria one of the most dreadful diseases affecting the quality of life lived by an individual in tropical countries.…”